NEISSERIAL ANTIGENS

Abstract: The invention provides proteins from Neisseria meningitidis (strains A and B) and from Neisseria gonorrhoerae including amino acid sequences, the corresponding nucleotide sequences, expression data, and serological data. The proteins are useful antigens for vaccines, immunogenic compositions, and/or diagnostics.

Description: NEISSERIAL ANTIGENS This invention relates to antigens from Neisseria bacteria. BACKGROUND ART Neisseria meningitidis .and Neisseria gonorrhoeae are non-motile, gram negative diplococci that are pathogenic in humans. N. meningitidis colonises the pharynx and causes meningitis (and, occasionally, septicaemia in the absence of meningitis); N.gonorrhoeae colonises the genital tract and causes gonorrhea. Although colonising different areas of the body and causing completely different diseases, the two pathogens are closely related, although one feature that clearly differentiates meningococcus from gonococcus is the presence of a polysaccharide capsule that is present in all pathogenic meningococci. N.gonorrhoeae caused approximately 800,000 cases per year during the period 1983-1990 in the United States alone (chapter by Meitzner & Cohen, "Vaccines Against Gonococcal Infection", In: New Generation Vaccines, 2nd edition, ed. Levine, Woodrow, Kaper, & Cobon, Marcel Dekker, New York, 1997, pp.817-842). The disease causes significant morbidity but limited mortality. Vaccination against N.gonorrhoeae would be highly desirable, but repeated attempts have failed. The main candidate antigens for this vaccine are surface-exposed proteins such as pili, porins, opacity-associated proteins (Opas) and other surface-exposed proteins such as the Lip, Laz, IgAl protease and transferrin-binding proteins. The lipooligosaccharide (LOS) has also been suggested as vaccine (Meitzner & Cohen, supra). N. meningitidis causes both endemic and epidemic disease. In the United States the attack rate is 0.6-1 per 100,000 persons per year, and it can be much greater during outbreaks (see Lieberman et al. (1996) Safety and Immunogenicity of a Serogroups A/C Neisseria meningitidis Oligosaccharide-Protein Conjugate Vaccine in Young Children. JAMA 275(19):1499-1503; Schuchat et al (1997) Bacterial Meningitis in the United States in 1995. NEnglJMed 337(14):970- 976). In developing countries, endemic disease rates are much higher and during epidemics incidence rates can reach 500 cases per 100,000 persons per year. Mortality is extremely high, at 10-20% in the United States, and much higher in developing countries. Following the introduction of the conjugate vaccine against Haemophilus inβuenzae, N meningitidis is the major cause of bacterial meningitis at all ages in the United States (Schuchat et al (1997) supra). Based on the organism's capsular polysaccharide, 12 serogroups of N meningitidis have been identified. Group A is the pathogen most often implicated in epidemic disease in sub-Saharan Africa. Serogroups B and C are responsible for the vast majority of cases in the United States and in most developed countries. Serogroups W135 and Y are responsible for the rest of the cases in the United States and developed countries. The meningococcal vaccine currently in use is a tetravalent polysaccharide vaccine composed of serogroups A, C, Y and W135. Although efficacious in adolescents and adults, it induces a poor immune response and short duration of protection, and cannot be used in infants [eg. Morbidity .and Mortality weekly report, Vol.46, No. RR-5 (1997)]. This is because polysaccharides are T-cell independent antigens that induce a weak immune response that cannot be boosted by repeated immunization. Following the success of the vaccination against H.influenzae, conjugate vaccines against serogroups A and C have been developed and are at the final stage of clinical testing (Zollinger WD "New and Improved Vaccines Against Meningococcal Disease" in: New Generation Vaccines, supra, pp. 469-488; Lieberman et al (1996) supra; Costantino et al (1992) Development and phase I clinical testing of a conjugate vaccine against meningococcus A and C. Vaccine 10:691-698). Meningococcus B remains a problem, however. This serotype currently is responsible for approximately 50% of total meningitis in the United States, Europe, and South America. The polysaccharide approach cannot be used because the menB capsular polysaccharide is a polymer of α(2-8)-linked N-acetyl neuraminic acid that is also present in mammalian tissue. This results in tolerance to the .antigen; indeed, if an immune response were elicited, it would be .anti-self, .and therefore undesirable. In order to avoid induction of autoimmunity and to induce a protective immune response, the capsular polysaccharide has, for instance, been chemically modified substituting the N-acetyl groups with Ν-propionyl groups, leaving the specific antigenicity unaltered (Romero & Outschoorn (1994) Current status of Meningococcal group B vaccine candidates: capsular or non-capsular? Clin Microbiol Rev 7(4):559-575). Alternative approaches to menB vaccines have used complex mixtures of outer membrane proteins (OMPs), containing either the OMPs alone, or OMPs enriched in porins, or deleted of the class 4 OMPs that are believed to induce antibodies that block bactericidal activity. This approach produces vaccines that are not well characterized. They are able to protect against the homologous str-ain, but .are not effective at large where there are many antigenic variants of the outer membrane proteins. To overcome the antigenic variability, multivalent vaccines containing up to nine different porins have been constructed (eg. Poolman JT (1992) Development of a meningococcal vaccine. Infect. Agents Dis. 4:13-28). Additional proteins to be used in outer membrane vaccines have been the opa and ope proteins, but none of these approaches have been able to overcome the antigenic variability (eg. Ala'Aldeen & Borriello (1996) The meningococcal transferrin-binding proteins 1 and 2 are both surface exposed and generate bactericidal antibodies capable of killing homologous and heterologous strains. Vaccine 14(l):49-53). A certain amount of sequence data is available for meningococcal and gonoccocal genes and proteins (eg. EP-A-0467714, WO96/29412), but this is by no means complete. The provision of further sequences could provide an opportunity to identify secreted or surface-exposed proteins that are presumed t.argets for the immune system and which are not antigenically variable. For instance, some of the identified proteins could be components of efficacious vaccines against meningococcus B, some could be components of vaccines against all meningococcal serotypes, and others could be components of vaccines against all pathogenic Neisseriae. THE INVENTION The invention provides proteins comprising the Neisserial amino acid sequences disclosed in the examples. These sequences relate to N meningitidis ox N.gonorrhoeae. It also provides proteins comprising sequences homologous (ie. having sequence identity) to the Neisserial amino acid sequences disclosed in the examples. Depending on the particul.ar sequence, the degree of identity is preferably greater than 50% (eg. 65%, 80%, 90%, or more). These homologous proteins include mutants and allelic variants of the sequences disclosed in the examples. Typically, 50% identity or more between two proteins is considered to be an indication of functional equivalence. Identity between the proteins is preferably determined by the Smith-Waterman homology search algorithm as implemented in the MPSRCH program (Oxford Molecular), using an affine gap search with parameters gap open penalty=12 and gap extension penalty=l . The invention further provides proteins comprising fragments of the Neisserial amino acid sequences disclosed in the examples. The fragments should comprise at least n consecutive .amino acids from the sequences and, depending on the particular sequence, n is 7 or more (eg. 8, 10, 12, 14, 16, 18, 20 or more). Preferably the fragments comprise an epitope from the sequence. The proteins of the invention can, of course, be prepared by various means (eg. recombinant expression, purification from cell culture, chemical synthesis etc.) and in various forms (eg. native, fusions etc.). They are preferably prepared in substantially pure or isolated form (ie. substantially free from other Neisserial or host cell proteins) According to a further .aspect, the invention provides antibodies which bind to these proteins. These may be polyclonal or monoclonal and may be produced by any suitable means. According to a further aspect, the invention provides nucleic acid comprising the Neisserial nucleotide sequences disclosed in the examples. In addition, the invention provides nucleic acid comprising sequences homologous (ie. having sequence identity) to the Neisserial nucleotide sequences disclosed in the examples. Furthermore, the invention provides nucleic acid which can hybridise to the Neisserial nucleic acid disclosed in the examples, preferably under "high stringency" conditions (eg. 65°C in a O.lxSSC, 0.5% SDS solution). Nucleic acid comprising fragments of these sequences are also provided. These should comprise at least n consecutive nucleotides from the Neisserial sequences and, depending on the particular sequence, n is 10 or more (eg 12, 14, 15, 18, 20, 25, 30, 35, 40 or more). According to a further aspect, the invention provides nucleic acid encoding the proteins and protein fragments of the invention. It should also be appreciated that the invention provides nucleic acid comprising sequences complementary to those described above (eg. for antisense or probing purposes). Nucleic acid according to the invention can, of course, be prepared in many ways (eg. by chemical synthesis, from genomic or cDNA libraries, from the organism itself etc.) and can take various forms (eg. single stranded, double stranded, vectors, probes etc.). In addition, the term "nucleic acid" includes DNA and RNA, and also their analogues, such as those containing modified backbones, and also peptide nucleic acids (PNA) etc. According to a further aspect, the invention provides vectors comprising nucleotide sequences of the invention (eg. expression vectors) and host cells transformed with such vectors. According to a further aspect, the invention provides compositions comprising protein, antibody, and/or nucleic acid according to the invention. These compositions may be suitable as vaccines, for instance, or as diagnostic reagents, or as immunogenic compositions. The invention also provides nucleic acid, protein, or antibody according to the invention for use as medicaments (eg. as vaccines) or as diagnostic reagents. It also provides the use of nucleic acid, protein, or antibody according to the invention in the manufacture of: (i) a medicament for treating or preventing infection due to Neisserial bacteria; (ii) a diagnostic reagent for detecting the presence of Neisserial bacteria or of antibodies raised against Neisserial bacteria; and/or (iii) a reagent which can raise antibodies against Neisserial bacteria. Said Neisserial bacteria may be any species or strain (such as N.gonorrhoeae, or any strain of N. meningitidis, such as strain A, strain B or strain C). The invention also provides a method of treating a patient, comprising administering to the patient a therapeutically effective amount of nucleic acid, protein, and/or antibody according to the invention. According to further aspects, the invention provides various processes. A process for producing proteins of the invention is provided, comprising the step of culturing a host cell according to the invention under conditions which induce protein expression. A process for producing protein or nucleic acid of the invention is provided, wherein the the protein or nucleic acid is synthesised in part or in whole using chemical means. A process for detecting polynucleotides of the invention is provided, comprising the steps of: (a) contacting a nucleic probe according to the invention with a biological sample under hybridizing conditions to form duplexes; and (b) detecting said duplexes. A process for detecting proteins of the invention is provided, comprising the steps of: (a) contacting an antibody according to the invention with a biological sample under conditions suitable for the formation of an antibody-antigen complexes; and (b) detecting said complexes. A summary of standard techniques and procedures which may be employed in order to perform the invention (eg. to utilise the disclosed sequences for vaccination or diagnostic purposes) follows. This summary is not a limitation on the invention but, rather, gives examples that may be used, but are not required. General The practice of the present invention will employ, unless otherwise indicated, conventional techniques of molecular biology, microbiology, recombinant DNA, and immunology, which are within the skill of the art. Such techniques are explained fully in the literature eg. Sambrook Molecular Cloning; A Laboratory Manual, Second Edition (1989); DNA Cloning, Volumes land IΪ (D.N Glover ed. 1985); Oligonucleotide Synthesis (M.J. Gait ed, 1984); Nucleic Acid Hybridization (B.D. Hames & S.J. Higgins eds. 1984); Transcription and Translation (B.D. Hames & S.J. Higgins eds. 1984); Animal Cell Culture (R.I. Freshney ed. 1986); Immobilized Cells and Enzymes (IRL Press, 1986); B. Perbal, A Practical Guide to Molecular Cloning (1984); the Methods in Enzymology series (Academic Press, Inc.), especially volumes 154 & 155; Gene Transfer Vectors for Mammalian Cells (J.H. Miller and M.P. Calos eds. 1987, Cold Spring Harbor Laboratory); Mayer and Walker, eds. (1987), Immunochemical Methods in Cell and Molecular Biology (Academic Press, London); Scopes, (1987) Protein Purification: Principles and Practice, Second Edition (Springer- Verlag, N.Y.), and Handbook of Experimental Immunology, Volumes I-IV (D.M. Weir and C. C. Blackwell eds 1986). Standard abbreviations for nucleotides and amino acids are used in this specification. All publications, patents, and patent applications cited herein are incorporated in full by reference. In particular, the contents of UK patent applications 9723516.2, 9724190.5, 9724386.9, 9725158.1, 9726147.3, 9800759.4, and 9819016.8 are incorporated herein. Definitions A composition containing X is "substantially free of Y when at least 85% by weight of the total X+Y in the composition is X. Preferably, X comprises at least about 90% by weight of the total of X+Y in the composition, more preferably at least about 95% or even 99% by weight. The term "comprising" means "including" as well as "consisting" eg. a composition "comprising" X may consist exclusively of X or may include something additional to X, such as X+Y. The term "heterologous" refers to two biological components that are not found together in nature. The components may be host cells, genes, or regulatory regions, such as promoters. Although the heterologous components .are not found together in nature, they can function together, as when a promoter heterologous to a gene is operably linked to the gene. Another example is where a Neisserial sequence is heterologous to a mouse host cell. A further examples would be two epitopes from the same or different proteins which have been assembled in a single protein in an arrangement not found in nature. An "origin of replication" is a polynucleotide sequence that initiates and regulates replication of polynucleotides, such as an expression vector. The origin of replication behaves as an autonomous unit of polynucleotide replication within a cell, capable of replication under its own control. An origin of replication may be needed for a vector to replicate in a particular host cell. With certain origins of replication, an expression vector can be reproduced at a high copy number in the presence of the appropriate proteins within the cell. Examples of origins are the autonomously replicating sequences, which are effective in yeast; and the viral T-antigen, effective in COS-7 cells. A "mutant" sequence is defined as DNA, RNA or amino acid sequence differing from but having sequence identity with the native or disclosed sequence. Depending on the particul.ar sequence, the degree of sequence identity between the native or disclosed sequence and the mutant sequence is preferably greater than 50% (eg. 60%, 70%, 80%, 90%, 95%, 99% or more, calculated using the Smith- Waterman algorithm as described above). As used herein, an "allelic variant" of a nucleic acid molecule, or region, for which nucleic acid sequence is provided herein is a nucleic acid molecule, or region, that occurs essentially at the same locus in the genome of another or second isolate, and that, due to natural variation caused by, for example, mutation or recombination, has a similar but not identical nucleic acid sequence. A coding region allelic variant typically encodes a protein having similar activity to that of the protein encoded by the gene to which it is being compared. An allelic variant can also comprise an alteration in the 5' or 3' untranslated regions of the gene, such as in regulatory control regions (eg. see US patent 5,753,235). Expression systems The Neisserial nucleotide sequences can be expressed in a variety of different expression systems; for example those used with mammalian cells, baculoviruses, plants, bacteria, and yeast. i. Mammalian Systems Mammalian expression systems are known in the art. A mammalian promoter is any DNA sequence capable of binding mammalian RNA polymerase and initiating the downstream (3') transcription of a coding sequence (eg. structural gene) into mRNA. A promoter will have a transcription initiating region, which is usually placed proximal to the 5' end of the coding sequence, and a TATA box, usually located 25-30 base pairs (bp) upstream of the transcription initiation site. The TATA box is thought to direct RNA polymerase II to begin RNA synthesis at the correct site. A mammalian promoter will also contain an upstream promoter element, usually located within 100 to 200 bp upstream of the TATA box. An upstream promoter element determines the rate at which transcription is imtiated and can act in either orientation [Sambrook et al. (1989) "Expression of Cloned Genes in Mammalian Cells." In Molecular Cloning: A Laboratory Manual, 2nd ed.]. Mammalian viral genes are often highly expressed and have a broad host range; therefore sequences encoding mammalian viral genes provide particularly useful promoter sequences. Examples include the SV40 early promoter, mouse mammary tumor virus LTR promoter, adenovirus major late promoter (Ad MLP), and herpes simplex virus promoter. In addition, sequences derived from non- viral genes, such as the murine metallotheionein gene, also provide useful promoter sequences. Expression may be either constitutive or regulated (inducible), depending on the promoter can be induced with glucocorticoid in hormone-responsive cells. The presence of an enhancer element (enhancer), combined with the promoter elements described above, will usually increase expression levels. An enhancer is a regulatory DNA sequence that can stimulate transcription up to 1000-fold when linked to homologous or heterologous promoters, with synthesis beginning at the normal RNA start site. Enhancers are also active when they are placed upstream or downstream from the transcription initiation site, in either normal or flipped orien- tation, or at a distance of more than 1000 nucleotides from the promoter [Maniatis et al. (1987) Science 236:1237; Alberts et al. (1989) Molecular Biology of the Cell, 2nd ed.]. Enhancer elements derived from viruses may be particularly useful, because they usually have a broader host range. Examples include the SV40 early gene enhancer [Dijkema et al (1985) EMBO J. 4:761] and the enhancer/promoters derived from the long terminal repeat (LTR) of the Rous Sarcoma Virus [Gorman et al. (1982b) Proc. Natl. Acad. Sci. 79:6777] and from human cytomegalovirus [Boshart et al. (1985) Cell 41:521}. Additionally, some enhancers are regulatable and become active only in the presence of an inducer, such as a hormone or metal ion [Sassone-Corsi and Borelli (1986) Trends Genet. 2:215; Maniatis et al. (1987) Science 236:1237]. A DNA molecule may be expressed intracellularly in mammalian cells. A promoter sequence may be directly linked with the DNA molecule, in which case the first amino acid at the N-terminus of the recombin.ant protein will always be a methionine, which is encoded by the ATG start codon. If desired, the N-terminus may be cleaved from the protein by in vitro incubation with cyanogen bromide. Alternatively, foreign proteins can also be secreted from the cell into the growth media by creating chimeric DNA molecules that encode a fusion protein comprised of a leader sequence fragment that provides for secretion of the foreign protein in mammalian cells. Preferably, there are processing sites encoded between the leader fragment and the foreign gene that can be cleaved either in vivo or in vitro. The leader sequence fragment usually encodes a signal peptide comprised of hydrophobic .amino acids which direct the secretion of the protein from the cell. The adenovirus triparite leader is an example of a leader sequence that provides for secretion of a foreign protein in mammalian cells. Usually, transcription termination and polyadenylation sequences recognized by mammalian cells are regulatory regions located 3' to the translation stop codon and thus, together with the promoter elements, flank the coding sequence. The 3' terminus of the mature mRNA is formed by site- specific post-transcriptional cleavage and polyadenylation [Birnstiel et al. (1985) Cell 41:349; Proudfoot and Whitelaw (1988) "Termination and 3' end processing of eukaryotic RNA. In Transcription and splicing (ed. B.D. Hames and D.M. Glover); Proudfoot (1989) Trends Biochem. Sci. 14:105]. These sequences direct the transcription of an mRNA which can be translated into the polypeptide encoded by the DNA. Examples of transcription terminater/polyadenylation signals include those derived from SV40 [Sambrook et al (1989) "Expression of cloned genes in cultured mammalian cells." In Molecular Cloning: A Laboratory Manual]. Usually, the above described components, comprising a promoter, polyadenylation signal, and transcription termination sequence are put together into expression constructs. Enhancers, introns with functional splice donor and acceptor sites, and leader sequences may also be included in an expression construct, if desired. Expression constructs are often maintained in a replicon, such as an extrachromosomal element (eg. plasmids) capable of stable maintenance in a host, such as mammalian cells or bacteria. Mammalian replication systems include those derived from animal viruses, which require trans-acting factors to replicate. For example, plasmids containing the replication systems of papovaviruses, such as SV40 [Gluzman (1981) Cell 25:175] or polyomavirus, replicate to extremely high copy number in the presence of the appropriate viral T antigen. Additional examples of mammalian replicons include those derived from bovine papillomavirus and Epstein-Barr virus. Additionally, the replicon may have two replicaton systems, thus allowing it to be maintained, for example, in mammalian cells for expression and in a prokaryotic host for cloning and amplification. Examples of such mammalian-bacteria shuttle vectors include pMT2 [Kaufman et al. (1989) Mol. Cell. Biol. 9:946] and pHEBO [Shimizu et al. (1986) Mol. Cell. Biol. 5:1074]. The transformation procedure used depends upon the host to be transformed. Methods for introduction of heterologous polynucleotides into mammalian cells are known in the art and include dextran-mediated transfection, calcium phosphate precipitation, polybrene mediated transfection, protoplast fusion, electroporation, encapsulation of the polynucleotide(s) in liposomes, and direct microinjection of the DNA into nuclei. Mammalian cell lines available as hosts for expression are known in the art and include many immortalized cell lines available from the American Type Culture Collection (ATCC), including but not limited to, Chinese hamster ovary (CHO) cells, HeLa cells, baby hamster kidney (BHK) cells, monkey kidney cells (COS), human hepatocellular carcinoma cells (eg. Hep G2), and a number of other cell lines. ii. Baculo virus Systems The polynucleotide encoding the protein can also be inserted into a suitable insect expression vector, and is operably linked to the control elements within that vector. Vector construction employs techniques which are known in the art. Generally, the components of the expression system include a transfer vector, usually a bacterial plasmid, which contains both a fragment of the baculovirus genome, and a convenient restriction site for insertion of the heterologous gene or genes to be expressed; a wild type baculovirus with a sequence homologous to the baculovirus-specific fragment in the transfer vector (this allows for the homologous recombination of the heterologous gene in to the baculovirus genome); and appropriate insect host cells and growth media. After inserting the DNA sequence encoding the protein into the transfer vector, the vector and the wild type viral genome are transfected into an insect host cell where the vector and viral genome are allowed to recombine. The packaged recombinant virus is expressed and recombinant plaques are identified and purified. Materials and methods for baculovirus/insect cell expression systems are commercially available in kit form from, ter alia, Invitrogen, San Diego CA ("MaxBac" kit). These techniques are generally known to those skilled in the art and fully described in Summers and Smith, Texas Agricultural Experiment Station Bulletin No. 1555 (1987) (hereinafter "Summers and Smith"). Prior to inserting the DNA sequence encoding the protein into the baculovirus genome, the above described components, comprising a promoter, leader (if desired), coding sequence of interest, and transcription termination sequence, are usually assembled into an intermediate transplacement construct (transfer vector). This construct may contain a single gene and operably linked regulatory elements; multiple genes, each with its owned set of operably linked regulatory elements; or multiple genes, regulated by the same set of regulatory elements. Intermediate transplacement constructs are often maintained in a replicon, such as an extrachromosomal element (eg. plasmids) capable of stable maintenance in a host, such as a bacterium. The replicon will have a replication system, thus allowing it to be maintained in a suitable host for cloning and amplification. Currently, the most commonly used transfer vector for introducing foreign genes into AcNPV is pAc373. Many other vectors, known to those of skill in the art, have also been designed. These include, for example, pVL985 (which alters the polyhedrin start codon from ATG to ATT, and which introduces a BamHl cloning site 32 basepairs downstream from the ATT; see Luckow and Summers, Virology (1989) 77:31. The pl.asmid usually also contains the polyhedrin polyadenylation signal (Miller et al. (1988) Ann. Rev. Microbiol, 42:177) and a prokaryotic ampicillin-resistance (amp) gene and origin of replication for selection and propagation in E. coli. Baculovirus transfer vectors usually contain a baculovirus promoter. A baculovirus promoter is any DNA sequence capable of binding a baculovirus RNA polymerase and initiating the downstream (5' to 3') transcription of a coding sequence (eg. structural gene) into mRNA. A promoter will have a transcription initiation region which is usually placed proximal to the 5' end of the coding sequence. This transcription initiation region usually includes an RNA polymerase binding site and a transcription initiation site. A baculovirus transfer vector may also have a second domain called an enhancer, which, if present, is usually distal to the structural gene. Expression may be either regulated or constitutive. Structural genes, abundantly transcribed at late times in a viral infection cycle, provide particularly useful promoter sequences. Examples include sequences derived from the gene encoding the viral polyhedron protein, Friesen et al., (1986) "The Regulation of Baculovirus Gene Expression," in: The Molecular Biology of Baculoviruses (ed. Walter Doerfler); EPO Publ. Nos. 127 839 and 155 476; and the gene encoding the plO protein, Vlak et al., (1988), J. Gen. Virol. 69:765. DNA encoding suitable signal sequences can be derived from genes for secreted insect or baculovirus proteins, such as the baculovirus polyhedrin gene (Carbonell et al. (1988) Gene, 73:409). Alternatively, since the signals for mammalian cell posttranslational modifications (such as signal peptide cleavage, proteolytic cleavage, and phosphorylation) appear to be recognized by insect cells, and the signals required for secretion and nuclear accumulation also appear to be conserved between the invertebrate cells and vertebrate cells, leaders of non-insect origin, such as those derived from genes encoding human α-interferon, Maeda et al., (1985), Nature 315:592; human gastrin-releasing peptide, Lebacq-Verheyden et al, (1988), Molec. Cell. Biol. 5:3129; human IL-2, Smith et al., (1985) Proc. Nat'lAcad. Sci. USA, #2:8404; mouse IL-3, (Miyajima et al., (1987) Gene 58:273; and human glucocerebrosidase, Martin et al. (1988) DNA, 7:99, can also be used to provide for secretion in insects. A recombinant polypeptide or polyprotein may be expressed intracellularly or, if it is expressed with the proper regulatory sequences, it can be secreted. Good intracellular expression of nonfused foreign proteins usually requires heterologous genes that ideally have a short leader sequence containing suitable translation initiation signals preceding an ATG start signal. If desired, methionine at the N-terminus may be cleaved from the mature protein by in vitro incubation with cyanogen bromide. Alternatively, recombinant polyproteins or proteins which are not naturally secreted can be secreted from the insect cell by creating chimeric DNA molecules that encode a fusion protein comprised of a leader sequence fragment that provides for secretion of the foreign protein in insects. The leader sequence fragment usually encodes a signal peptide comprised of hydrophobic amino acids which direct the translocation of the protein into the endoplasmic reticulum. After insertion of the DNA sequence and/or the gene encoding the expression product precursor of the protein, an insect cell host is co-transformed with the heterologous DNA of the transfer vector and the genomic DNA of wild type baculovirus — usually by co-transfection. The promoter and transcription termination sequence of the construct will usually comprise a 2-5kb section of the baculovirus genome. Methods for introducing heterologous DNA into the desired site in the baculovirus virus are known in the art. (See Summers and Smith supra; Ju et al. (1987); Smith et al., Mol. Cell. Biol. (1983) 5:2156; .and Luckow .and Summers (1989)). For example, the insertion can be into a gene such as the polyhedrin gene, by homologous double crossover recombination; insertion can also be into a restriction enzyme site engineered into the desired baculovirus gene. Miller et al, (1989), Bioessays :91.The DNA sequence, when cloned in place of the polyhedrin gene in the expression vector, is flanked both 5' and 3' by polyhedrin-specific sequences and is positioned downstream of the polyhedrin promoter. The newly formed baculovirus expression vector is subsequently packaged into an infectious recombinant baculovirus. Homologous recombination occurs at low frequency (between about 1% and about 5%); thus, the majority of the virus produced after cotransfection is still wild-type virus. Therefore, a method is necessary to identify recombinant viruses. An advantage of the expression system is a visual screen allowing recombinant viruses to be distinguished. The polyhedrin protein, which is produced by the native virus, is produced at very high levels in the nuclei of infected cells at late times after viral infection. Accumulated polyhedrin protein forms occlusion bodies that also contain embedded particles. These occlusion bodies, up to 15 μm in size, are highly refractile, giving them a bright shiny appearance that is readily visualized under the light microscope. Cells infected with recombinant viruses lack occlusion bodies. To distinguish recombinant virus from wild-type virus, the transfection supernatant is plaqued onto a monolayer of insect cells by techniques known to those skilled in the art. Namely, the plaques are screened under the light microscope for the presence (indicative of wild-type virus) or absence (indicative of recombinant virus) of occlusion bodies. "Current Protocols in Microbiology" Vol. 2 (Ausubel et al. eds) at 16.8 (Supp. 10, 1990); Summers and Smith, supra; Miller et al. (1989). Recombinant baculovirus expression vectors have been developed for infection into several insect cells. For example, recombinant baculoviruses have been developed for, inter alia: Aedes aegypti , Autographa californica, Bombyx mori, Drosophila melanogaster, Spodoptera frugiperda, and Trichoplusiα ni (WO 89/046699; Garbonell et al, (1985) J Virol. 56:153; Wright (1986) Nαtwre 527:718; Smith et al, (1983) Mol. Cell. Biol. 5:2156; and see generally, Fraser, et αl. (1989) In Vitro Cell. Dev. Biol. 25:225). Cells and cell culture media are commercially available for both direct and fusion expression of heterologous polypeptides in a baculovirus/expression system; cell culture technology is generally known to those skilled in the art. See, eg. Summers and Smith supra. The modified insect cells may then be grown in an appropriate nutrient medium, which allows for stable maintenance of the plasmid(s) present in the modified insect host. Where the expression product gene is under inducible control, the host may be grown to high density, and expression induced. Alternatively, where expression is constitutive, the product will be continuously expressed into the medium and the nutrient medium must be continuously circulated, while removing the product of interest and augmenting depleted nutrients. The product may be purified by such techniques as chromatography, eg. HPLC, affinity chromatography, ion exchange chromatography, etc.; electrophoresis; density gradient centrifugation; solvent extraction, or the like. As appropriate, the product may be further purified, as required, so as to remove substantially any insect proteins which are also secreted in the medium or result from lysis of insect cells, so as to provide a product which is at least substantially free of host debris, eg. proteins, lipids and polysaccharides. In order to obtain protein expression, recombinant host cells derived from the transformants are incubated under conditions which allow expression of the recombinant protein encoding sequence. These conditions will vary, dependent upon the host cell selected. However, the conditions are readily ascertainable to those of ordinary skill in the art, based upon what is known in the art. iii. Plant Systems There are many plant cell culture and whole plant genetic expression systems known in the art. Exemplary plant cellular genetic expression systems include those described in patents, such as: US 5,693,506; US 5,659,122; and US 5,608,143. Additional examples of genetic expression in plant cell culture has been described by Zenk, Phytochemistry 30:3861-3863 (1991). Descriptions of plant protein signal peptides may be found in addition to the references described above in Vaulcombe et al., Mol. Gen. Genet. 209:33-40 (1987); Chandler et al., Plant Molecular Biology 3:407-418 (1984); Rogers, J Biol. Chem. 260:3731-3738 (1985); Rothstein et al., Gene 55:353-356 (1987); Whittier et al., Nucleic Acids Research 15:2515-2535 (1987); Wirsel et al., Molecular Microbiology 3:3-14 (1989); Yu et al., Gene 122:247-253 (1992). A description of the regulation of plant gene expression by the phytohormone, gibberellic acid and secreted enzymes induced by gibberellic acid can be found in R.L. Jones and J. MacMillin, Gibberellins: in: Advanced Plant Physiology,. Malcolm B. Wilkins, ed., 1984 Pitman Publishing Limited, London, pp. 21-52. References that describe other metabolically-regulated genes: Sheen, Plant Cell, 2:1027- 1038(1990); Maas et al., EMBOJ. 9:3447-3452 (1990); Benkel and Hickey, Proc. Natl. Acad. Sci. 84:1337-1339 (1987) Typically, using techniques known in the art, a desired polynucleotide sequence is inserted into an expression cassette comprising genetic regulatory elements designed for operation in plants. The expression cassette is inserted into a desired expression vector with companion sequences upstream and downstream from the expression cassette suitable for expression in a plant host. The companion sequences will be of plasmid or viral origin and provide necessary characteristics to the vector to permit the vectors to move DNA from .an original cloning host, such as bacteria, to the desired plant host. The basic bacterial/plant vector construct will preferably provide a broad host range prokaryote replication origin; a prokaryote selectable marker; and, for Agrobacterium transformations, T DNA sequences for Agrobacterium-mediated transfer to plant chromosomes. Where the heterologous gene is not readily amenable to detection, the construct will preferably also have a selectable marker gene suitable for determining if a plant cell has been transformed. A general review of suitable markers, for example for the members of the grass family, is found in Wilmink and Dons, 1993, Plant Mol. Biol. Reptr, 11(2):165-185. Sequences suitable for permitting integration of the heterologous sequence into the plant genome are also recommended. These might include transposon sequences and the like for homologous recombination as well as Ti sequences which permit random insertion of a heterologous expression cassette into a plant genome. Suitable prokaryote selectable markers include resistance toward antibiotics such as ampicillin or tetracycline. Other DNA sequences encoding additional functions may also be present in the vector, as is known in the art. The nucleic acid molecules of the subject invention may be included into an expression cassette for expression of the protein(s) of interest. Usually, there will be only one expression cassette, although two or more are feasible. The recombinant expression cassette will contain in addition to the heterologous protein encoding sequence the following elements, a promoter region, plant 5' untranslated sequences, initiation codon depending upon whether or not the structural gene comes equipped with one, and a transcription .and translation termination sequence. Unique restriction enzyme sites at the 5' and 3' ends of the cassette allow for easy insertion into a pre-existing vector. A heterologous coding sequence may be for any protein relating to the present invention. The sequence encoding the protein of interest will encode a signal peptide which allows processing and translocation of the protein, as appropriate, and will usually lack any sequence which might result in the binding of the desired protein of the invention to a membrane. Since, for the most part, the transcriptional initiation region will be for a gene which is expressed and translocated during germination, by employing the signal peptide which provides for translocation, one may also provide for translocation of the protein of interest. In this way, the protein(s) of interest will be translocated from the cells in which they are expressed and may be efficiently harvested. Typically secretion in seeds are across the aleurone or scutellar epithelium layer into the endosperm of the seed. While it is not required that the protein be secreted from the cells in which the protein is produced, this facilitates the isolation and purification of the recombinant protein. Since the ultimate expression of the desired gene product will be in a eucaryotic cell it is desirable to determine whether any portion of the cloned gene contains sequences which will be processed out as introns by the host's splicosome machinery. If so, site-directed mutagenesis of the "intron" region may be conducted to prevent losing a portion of the genetic message as a false intron code, Reed and Maniatis, Cell 41:95-105, 1985. The vector can be microinjected directly into plant cells by use of micropipettes to mechanically transfer the recombinant DNA. Crossway, Mol. Gen. Genet, 202:179-185, 1985. The genetic material may also be transferred into the plant cell by using polyethylene glycol, Krens, et al., Nature, 296, 72-7 , 1982. Another method of introduction of nucleic acid segments is high velocity ballistic penetration by small particles with the nucleic acid either within the matrix of small beads or particles, or on the surface, Klein, et al., Nature, 327, 70-73, 1987 and Knudsen and Muller, 1991, Planta, 185:330-336 teaching particle bombardment of barley endosperm to create transgenic barley. Yet another method of introduction would be fusion of protoplasts with other entities, either minicells, cells, lysosomes or other fusible lipid-surfaced bodies, Fraley, et al., Proc. Natl. Acad. Sci. USA, 79, 1859-1863, 1982. The vector may also be introduced into the plant cells by electroporation. (Fromm et al., Proc. Natl Acad. Sci. USA 82:5824, 1985). In this technique, plant protoplasts are electroporated in the presence of plasmids containing the gene construct. Electrical impulses of high field strength reversibly permeabilize biomembranes allowing the introduction of the plasmids. Electroporated plant protoplasts reform the cell wall, divide, and form plant callus. All plants from which protoplasts can be isolated and cultured to give whole regenerated plants can be transformed by the present invention so that whole plants are recovered which contain the transferred gene. It is known that practically all plants can be regenerated from cultured cells or tissues, including but not limited to all major species of sugarcane, sugar beet, cotton, fruit and other trees, legumes and vegetables. Some suitable plants include, for example, species from the genera Fragaria, Lotus, Medicago, Onobrychis, Trifolium, Trigonella, Vigna, Citrus, Linum, Geranium, Manihot, Daucus, Arabidopsis, Brassica, Raphanus, Sinapis, Atropa, Capsicum, Datura, Hyoscyamus, Lycopersion, Nicotiana, Solanum, Petunia, Digitalis, Majorana, Cichorium, Helianthus, Lactuca, Bromus, Asparagus, Antirrhinum, Hererocallis, Nemesia, Pelargonium, Panicum, Pennisetum, Ranunculus, Senecio, Salpiglossis, Cucumis, Browaalia, Glycine, Lolium, Zea, Triticum, Sorghum, .and Datura. Means for regeneration vary from species to species of plants, but generally a suspension of transformed protoplasts containing copies of the heterologous gene is first provided. Callus tissue is formed and shoots may be induced from callus and subsequently rooted. Alternatively, embryo formation can be induced from the protoplast suspension. These embryos germinate as natural embryos to form plants. The culture media will generally contain various amino acids and hormones, such as auxin and cytokinins. It is also advantageous to add glutamic acid and proline to the medium, especially for such species as corn and alfalfa. Shoots and roots normally develop simultaneously. Efficient regeneration will depend on the medium, on the genotype, and on the history of the culture. If these three variables are controlled, then regeneration is fully reproducible and repeatable. In some plant cell culture systems, the desired protein of the invention may be excreted or alternatively, the protein may be extracted from the whole plant. Where the desired protein of the invention is secreted into the medium, it may be collected. Alternatively, the embryos and embryoless-half seeds or other plant tissue may be mechanically disrupted to release any secreted protein between cells and tissues. The mixture may be suspended in a buffer solution to retrieve soluble proteins. Conventional protein isolation and purification methods will be then used to purify the recombinant protein. Parameters of time, temperature pH, oxygen, and volumes will be adjusted through routine methods to optimize expression and recovery of heterologous protein. iv. Bacterial Systems Bacterial expression techniques are known in the art. A bacterial promoter is any DNA sequence capable of binding bacterial RNA polymerase and initiating the downstream (3') transcription of a coding sequence (eg. structural gene) into mRNA. A promoter will have a transcription initiation region which is usually placed proximal to the 5' end of the coding sequence. This transcription initiation region usually includes an RNA polymerase binding site and a transcription initiation site. A bacterial promoter may also have a second domain called an operator, that may overlap an adjacent RNA polymerase binding site at which RNA synthesis begins. The operator permits negative regulated (inducible) transcription, as a gene repressor protein may bind the operator and thereby inhibit transcription of a specific gene. Constitutive expression may occur in the absence of negative regulatory elements, such as the operator. In addition, positive regulation may be achieved by a gene activator protein binding sequence, which, if present is usually proximal (5') to the RNA polymerase binding sequence. An example of a gene activator protein is the catabolite activator protein (CAP), which helps initiate transcription of the lac operon in Escherichia coli (E. coli) [Raibaud et al. (1984) Annu. Rev. Genet. 18:173]. Regulated expression may therefore be either positive or negative, thereby either enhancing or reducing transcription. Sequences encoding metabolic pathway enzymes provide particularly useful promoter sequences. Examples include promoter sequences derived from sugar metabolizing enzymes, such as galactose, lactose (lac) [Chang et al. (1977) Nature 795:1056], and maltose. Additional examples include promoter sequences derived from biosynthetic enzymes such as tryptophan (trp) [Goeddel et al. (1980) NMC. Acids Res. 5:4057; Yelverton et al. (1981) Nucl. Acids Res. 9:731; US patent 4,738,921; EP-A-0036776 and EP-A-0121775]. The g-laotamase (bid) promoter system [Weissmann (1981) "The cloning of interferon and other mistakes." In Interferon 3 (ed. I. Gresser)], bacteriophage lambda PL [Shimatake et al. (1981) Nature 292:128] and T5 [US patent 4,689,406] promoter systems also provide useful promoter sequences. In addition, synthetic promoters which do not occur in nature also function as bacterial promoters. For example, transcription activation sequences of one bacterial or bacteriophage promoter may be joined with the operon sequences of another bacterial or bacteriophage promoter, creating a synthetic hybrid promoter [US patent 4,551,433]. For example, the tac promoter is a hybrid trp-lac promoter comprised of both trp promoter .and lac operon sequences that is regulated by the lac repressor [A ann et al. (1983) Gene 25:167; de Boer et al. (1983) Proc. Natl. Acad. Sci. 80:21]. Furthermore, a bacterial promoter can include naturally occurring promoters of non-bacterial origin that have the ability to bind bacterial RNA polymerase and initiate transcription. A naturally occurring promoter of non-bacterial origin can also be coupled with a compatible RNA polymerase to produce high levels of expression of some genes in prokaryotes. The bacteriophage T7 RNA polymerase/promoter system is an example of a coupled promoter system [Studier et al. (1986) J. Mol. Biol. 759:113; Tabor et al. (1985) Proc Natl. Acad. Sci. 52:1074]. In addition, a hybrid promoter can also be comprised of a bacteriophage promoter and an E. coli operator region (ΕPO- A-0 267 851). In addition to a functioning promoter sequence, an efficient ribosome binding site is also useful for the expression of foreign genes in prokaryotes. In E. coli, the ribosome binding site is called the Shine-Dalgarno (SD) sequence and includes an initiation codon (ATG) and a sequence 3-9 nucleotides in length located 3-11 nucleotides upstream of the initiation codon [Shine et al. (1975) Nature 254:3A]. The SD sequence is thought to promote binding of mRNA to the ribosome by the pairing of bases between the SD sequence and the 3' and of E. coli 16S rRNA [Steitz et al. (1979) "Genetic signals and nucleotide sequences in messenger RNA." In Biological Regulation and Development: Gene Expression (ed. R.F. Goldberger)]. To express eukaryotic genes and prokaryotic genes with weak ribosome-binding site [Sambrook et al. (1989) "Expression of cloned genes in Escherichia coli." In Molecular Cloning: A Laboratory Manual]. A DNA molecule may be expressed infracellularly. A promoter sequence may be directly linked with the DNA molecule, in which case the first amino acid at the N-terminus will always be a methionine, which is encoded by the ATG start codon. If desired, methionine at the N-terminus may be cleaved from the protein by in vitro incubation with cyanogen bromide or by either in vivo on in vitro incubation with a bacterial methionine N-terminal peptidase (EPO-A-0 219 237). Fusion proteins provide an alternative to direct expression. Usually, a DNA sequence encoding the N-terminal portion of an endogenous bacterial protein, or other stable protein, is fused to the 5' end of heterologous coding sequences. Upon expression, this construct will provide a fusion of the two amino acid sequences. For example, the bacteriophage lambda cell gene can be linked at the 5' terminus of a foreign gene and expressed in bacteria. The resulting fusion protein preferably retains a site for a processing enzyme (factor Xa) to cleave the bacteriophage protein from the foreign gene [Nagai et al. (1984) Nature 509:810]. Fusion proteins can also be made with sequences from the lacZ [Jia et al. (1987) Gene 60:197], trpE [Allen et al. (1987) j. Biotechnol. 5:93; Makoff et al. (1989) J. Gen. Microbiol. 135:11], and Chey [EP-A-0 324 647] genes. The DNA sequence at the junction of the two amino acid sequences may or may not encode a cleavable site. Another example is a ubiquitin fusion protein. Such a fusion protein is made with the ubiquitin region that preferably retains a site for a processing enzyme (eg. ubiquitin specific processing-protease) to cleave the ubiquitin from the foreign protein. Through this method, native foreign protein can be isolated [Miller et al. (1989) Bio/Technology 7:698]. Alternatively, foreign proteins can also be secreted from the cell by creating chimeric DNA molecules that encode a fusion protein comprised of a signal peptide sequence fragment that provides for secretion of the foreign protein in bacteria [US patent 4,336,336]. The signal sequence fragment usually encodes a signal peptide comprised of hydrophobic amino acids which direct the secretion of the protein from the cell. The protein is either secreted into the growth media (gram-positive bacteria) or into the periplasmic space, located between the inner and outer membrane of the cell (gram-negative bacteria). Preferably there are processing sites, which can be cleaved either in vivo or in vitro encoded between the signal peptide fragment and the foreign gene. DNA encoding suitable signal sequences can be derived from genes for secreted bacterial proteins, such as the E. coli outer membrane protein gene (ompA) [Masui et al. (1983), in: Experimental Manipulation of Gene Expression; Ghrayeb et al. (1984) EMBO J. 5:2437] and the E. coli alkaline phosphatase signal sequence (phoA) [Oka et al. (1985) Proc. Natl. Acad. Sci. 52:7212]. As an additional example, the signal sequence of the alpha-amylase gene from various Bacillus strains can be used to secrete heterologous proteins from B. subtilis [Palva et al. (1982) Proc. Natl. Acad. Sci. USA 79:5582; EP-A-0 244 042]. Usually, transcription termination sequences recognized by bacteria are regulatory regions located 3' to the translation stop codon, and thus together with the promoter flank the coding sequence. These sequences direct the transcription of an mRNA which can be translated into the polypeptide encoded by the DNA. Transcription termination sequences frequently include DNA sequences of about 50 nucleotides capable of forming stem loop structures that aid in terminating transcription. Examples include transcription termination sequences derived from genes with strong promoters, such as the trp gene in E. coli as well as other biosynthetic genes. Usually, the above described components, comprising a promoter, signal sequence (if desired), coding sequence of interest, and transcription termination sequence, are put together into expression constructs. Expression constructs are often maintained in a replicon, such as an extrachromosomal element (eg. plasmids) capable of stable maintenance in a host, such as bacteria. The replicon will have a replication system, thus allowing it to be maintained in a prokaryotic host either for expression or for cloning and amplification. In addition, a replicon may be either a high or low copy number plasmid. A high copy number plasmid will generally have a copy number ranging from about 5 to about 200, and usually about 10 to about 150. A host containing a high copy number plasmid will preferably contain at least about 10, and more preferably at least about 20 plasmids. Either a high or low copy number vector may be selected, depending upon the effect of the vector and the foreign protein on the host. Alternatively, the expression constructs can be integrated into the bacterial genome with an integrating vector. Integrating vectors usually contain at least one sequence homologous to the bacterial chromosome that allows the vector to integrate. Integrations appear to result from recombinations between homologous DNA in the vector and the bacterial chromosome. For example, integrating vectors constructed with DNA from various Bacillus strains integrate into the Bacillus chromosome (EP-A- 0 127 328). Integrating vectors may also be comprised of bacteriophage or transposon sequences. Usually, extrachromosomal and integrating expression constructs may contain selectable markers to allow for the selection of bacterial strains that have been transformed. Selectable markers can be expressed in the bacterial host and may include genes which render bacteria resistant to drugs such as ampicillin, chloramphenicol, erythromycin, kanamycin (neomycin), and tetracycline [Davies et al. (1978) Annu. Rev. Microbiol. 52:469]. Selectable markers may also include biosynthetic genes, such as those in the histidine, tryptophan, and leucine biosynthetic pathways. Alternatively, some of the above described components can be put together in transformation vectors. Transformation vectors are usually comprised of a selectable market that is either maintained in a replicon or developed into an integrating vector, as described above. Expression and transformation vectors, either extra-chromosomal replicons or integrating vectors, have been developed for transformation into many bacteria. For example, expression vectors have been developed for, inter alia, the following bacteria: Bacillus subtilis [Palva et al. (1982) Proc. Natl. Acad. Sci. USA 79:5582; EP-A-0 036 259 and EP-A-0 063 953; WO 84/04541], Escherichia coli [Shimatake et al. (1981) Nαtwre 292:128; Amann et al. (1985) Gene 40:183; Studier et al. (1986) J Mol. Biol. 759:113; EP-A-0 036 776,EP-A-0 136 829 and EP-A-0 136 907], Streptococcus cremoris [Powell et al. (1988) Appl. Environ. Microbiol. 54:655]; Streptococcus lividans [Powell et al. (1988) Appl. Environ. Microbiol. 54:655], Streptomyces lividans [US patent 4,745,056]. Methods of introducing exogenous DNA into bacterial hosts are well-known in the art, and usually include either the transformation of bacteria treated with CaCl 2 or other agents, such as divalent cations and DMSO. DNA can also be introduced into bacterial cells by electroporation. Transformation procedures usually vary with the bacterial species to be transformed. See eg. [Masson et al. (1989) FEMS Microbiol. Lett. 60:273; Palva et al. (1982) Proc. Natl. Acad. Sci. USA 79:5582; EP-A-0 036 259 and EP-A-0 063 953; WO 84/04541, Bacillus], [Miller et al. (1988) Proc. Natl. Acad. Sci. 55:856; Wang et al. (1990) J. Bacteriol. 172:949, Campylobacter], [Cohen et al. (1973) Proc. Natl. Acad. Sci. 59:2110; Dower et al. (1988) Nucleic Acids Res. 16:6127; Kushner (1978) "An improved method for transformation of Escherichia coli with ColEl-derived plasmids. In Genetic Engineering: Proceedings of the International Symposium on Genetic Engineering (eds. H.W. Boyer and S. Nicosia); Mandel et al. (1970) J. Mol. Biol. 55:159; Taketo (1988) Biochim. Biophys. Acta 949:318; Escherichia], [Chassy et al. (1987) FEMS Microbiol. Lett. 44: 73 Lactobacillus]; [Fiedler et al. (1988) Anal. Biochem 170:38, Pseudomonas]; [Augustin et al. (1990) FEMS Microbiol. Lett. 66:203, Staphylococcus], [Barany et al. (1980) J. Bacteriol. 144:698; Harlander (1987) "Transformation of Streptococcus lactis by electroporation, in: Streptococcal Genetics (ed. J. Ferretti and R. Curtiss III); Perry et al. (1981) Infect. Immun. 52:1295; Powell et al. (1988) Appl. Environ. Microbiol. 54:655; Somkuti et al. (1987) Proc. 4th Evr. Cong. Biotechnology 7:412, Streptococcus]. v. Yeast Expression Yeast expression systems are also known to one of ordinary skill in the art. A yeast promoter is any DNA sequence capable of binding yeast RNA polymerase and initiating the downstream (3') transcription of a coding sequence (eg. structural gene) into mRNA. A promoter will have a transcription initiation region which is usually placed proximal to the 5' end of the coding sequence. This transcription initiation region usually includes an RNA polymerase binding site (the "TATA Box") and a transcription initiation site. A yeast promoter may also have a second domain called an upstream activator sequence (UAS), which, if present, is usually distal to the structural gene. The UAS permits regulated (inducible) expression. Constitutive expression occurs in the absence of a UAS. Regulated expression may be either positive or negative, thereby either enhancing or reducing transcription. Yeast is a fermenting organism with an active metabolic pathway, therefore sequences encoding enzymes in the metabolic pathway provide particularly useful promoter sequences. Examples include alcohol dehydrogenase (ADH) (EP-A-0 284 044), enolase, glucokinase, glucose-6- phosphate isomerase, glyceraldehyde-3-phosphate-dehydrogenase (GAP or GAPDH), hexokinase, phosphofructokinase, 3-phosphoglycerate mutase, and pyruvate kinase (PyK) (EPO-A-0 329 203). The yeast PH05 gene, encoding acid phosphatase, also provides useful promoter sequences [Myanohara et al. (1983) Proc. Natl. Acad. Sci. USA 80:1]. In addition, synthetic promoters which do not occur in nature also function as yeast promoters. For example, UAS sequences of one yeast promoter may be joined with the transcription activation region of another yeast promoter, creating a synthetic hybrid promoter. Examples of such hybrid promoters include the ADH regulatory sequence linked to the GAP transcription activation region (US Patent Nos. 4,876,197 and 4,880,734). Other examples of hybrid promoters include promoters which consist of the regulatory sequences of either the ADH2, GAL4, GALIO, OR PH05 genes, combined with the transcriptional activation region of a glycolytic enzyme gene such as GAP or PyK (EP-A-0 164 556). Furthermore, a yeast promoter can include naturally occurring promoters of non-yeast origin that have the ability to bind yeast RNA polymerase and initiate transcription. Examples of such promoters include, inter alia, [Cohen et al. (1980) Proc. Natl. Acad. Sci. USA 77:1078; Henikoff et al. (1981) Nαtwre 255:835; Hollenberg et al. (1981) Curr. Topics Microbiol. Immunol. 96:1 9; Hollenberg et al. (1979) "The Expression of Bacterial Antibiotic Resistance Genes in the Yeast Saccharomyces cerevisiae," in: Plasmids of Medical, Environmental and Commercial Importance (eds. K.Ν. Timmis and A. Puhler); Mercerau-Puigalon et al. (1980) Gene 77:163; Panthier et al. (1980) Curr. Genet. 2:109;]. A DNA molecule may be expressed mtracellularly in yeast. A promoter sequence may be directly linked with the DNA molecule, in which case the first amino acid at the N-terminus of the recombinant protein will always be a methionine, which is encoded by the ATG start codon. If desired, methionine at the N-terminus may be cleaved from the protein by in vitro incubation with cyanogen bromide. Fusion proteins provide an alternative for yeast expression systems, as well as in mammalian, baculovirus, and bacterial expression systems. Usually, a DNA sequence encoding the N-terminal portion of an endogenous yeast protein, or other stable protein, is fused to the 5' end of heterologous coding sequences. Upon expression, this construct will provide a fusion of the two amino acid sequences. For example, the yeast or human superoxide dismutase (SOD) gene, can be linked at the 5' terminus of a foreign gene and expressed in yeast. The DNA sequence at the junction of the two amino acid sequences may or may not encode a cleavable site. See eg. EP-A-0 196 056. Another example is a ubiquitin fusion protein. Such a fusion protein is made with the ubiquitin region that preferably retains a site for a processing enzyme (eg. ubiquitin-specific processing protease) to cleave the ubiquitin from the foreign protein. Through this method, therefore, native foreign protein can be isolated (eg. WO88/024066). Alternatively, foreign proteins can also be secreted from the cell into the growth media by creating chimeric DNA molecules that encode a fusion protein comprised of a leader sequence fragment that provide for secretion in yeast of the foreign protein. Preferably, there are processing sites encoded between the leader fragment and the foreign gene that can be cleaved either in vivo or in vitro. The leader sequence fragment usually encodes a signal peptide comprised of hydrophobic amino acids which direct the secretion of the protein from the cell. DNA encoding suitable signal sequences can be derived from genes for secreted yeast proteins, such as the yeast invertase gene (EP-A-0 012 873; JPO. 62,096,086) and the A-factor gene (US patent 4,588,684). Alternatively, leaders of non-yeast origin, such as an interferon leader, exist that also provide for secretion in yeast (EP-A-0 060 057). A preferred class of secretion leaders are those that employ a fragment of the yeast alpha-factor gene, which contains both a "pre" signal sequence, and a "pro" region. The types of alpha-factor fragments that can be employed include the full-length pre-pro alpha factor leader (about 83 amino acid residues) as well as truncated alpha-factor leaders (usually about 25 to about 50 amino acid residues) (US Patents 4,546,083 and 4,870,008; EP-A-0 324 274). Additional leaders employing an alpha-factor leader fragment that provides for secretion include hybrid alpha-factor leaders made with a presequence of a first yeast, but a pro-region from a second yeast alphafactor. (eg. see WO 89/02463.) Usually, transcription termination sequences recognized by yeast are regulatory regions located 3' to the translation stop codon, and thus together with the promoter flank the coding sequence. These sequences direct the transcription of an mRNA which can be translated into the polypeptide encoded by the DNA. Examples of transcription terminator sequence and other yeast-recognized termination sequences, such as those coding for glycolytic enzymes. Usually, the above described components, comprising a promoter, leader (if desired), coding sequence of interest, and transcription termination sequence, are put together into expression constructs. Expression constructs are often maintained in a replicon, such as an extrachromosomal element (eg. plasmids) capable of stable maintenance in a host, such as yeast or bacteria. The replicon may have two replication systems, thus allowing it to be maintained, for example, in yeast for expression and in a prokaryotic host for cloning and amplification. Examples of such yeast- bacteria shuttle vectors include YEp24 [Botstein et al. (1979) Gene 5:17-24], pCl/1 [Brake et al. (1984) Proc. Natl. Acad. Sci USA 57:4642-4646], and YRpl7 [Stinchcomb et al. (1982) J. Mol. Biol. 755:157]. In addition, a replicon may be either a high or low copy number plasmid. A high copy number plasmid will generally have a copy number ranging from about 5 to about 200, and usually about 10 to about 150. A host containing a high copy number plasmid will preferably have at least about 10, and more preferably at least about 20. Enter a high or low copy number vector may be selected, depending upon the effect of the vector and the foreign protein on the host. See eg. Brake et al., supra. Alternatively, the expression constructs can be integrated into the yeast genome with an integrating vector. Integrating vectors usually contain at least one sequence homologous to a yeast chromosome that allows the vector to integrate, and preferably contain two homologous sequences flanking the expression construct. Integrations appear to result from recombinations between homologous DNA in the vector and the yeast chromosome [Orr- Weaver et al. (1983) Methods in Enzymol. 101:228-245]. An integrating vector may be directed to a specific locus in yeast by selecting the appropriate homologous sequence for inclusion in the vector. See Orr- Weaver et al., supra. One or more expression construct may integrate, possibly affecting levels of recombinant protein produced [Rine et al. (1983) Proc. Natl. Acad. Sci. USA 80:6750]. The chromosomal sequences included in the vector can occur either as a single segment in the vector, which results in the integration of the entire vector, or two segments homologous to adjacent segments in the chromosome and flanking the expression construct in the vector, which can result in the stable integration of only the expression construct. Usually, extrachromosomal and integrating expression constructs may contain selectable markers to allow for the selection of yeast strains that have been transformed. Selectable markers may include biosynthetic genes that can be expressed in the yeast host, such as ADE2, HIS4, LEU2, TRP1, and ALG7, and the G418 resistance gene, which confer resistance in yeast cells to tunicamycin and G418, respectively. In addition, a suitable selectable marker may also provide yeast with the ability to grow in the presence of toxic compounds, such as metal. For example, the presence of CUP1 allows yeast to grow in the presence of copper ions [Butt et al. (1987) Microbiol, Rev. 57:351]. Alternatively, some of the above described components can be put together into transformation vectors. Transformation vectors are usually comprised of a selectable marker that is either maintained in a replicon or developed into an integrating vector, as described above. Expression and transformation vectors, either extrachromosomal replicons or integrating vectors, have been developed for transformation into many yeasts. For example, expression vectors have been developed for, inter alia, the following yeasts: Candida albicans [Kurtz, et al. (1986) Mol. Cell. Biol. .5:142], Candida maltosa [Kunze, et al. (1985) J. Basic Microbiol. 25:141]. Hansenula polymorpha [Gleeson, et al. (1986) J. Gen. Microbiol. 132:3459; Roggenkamp et al. (1986) Mol. Gen. Genet. 202:302], Kluyveromyces fragilis [Das, et al. (1984) J. Bacteriol. 755:1165], Kluyveromyces lactis [De Louvencourt et al. (1983) J. Bacteriol. 154:737; Van den Berg et al. (1990) Bio/Technology 5:135], Pichia guillerimondii [Kunze et al. (1985) J. Basic Microbiol. 25:141], Pichia pastoris [Cregg, et al. (1985) Mol. Cell. Biol. 5:3376; US Patent Nos. 4,837,148 and 4,929,555], Saccharomyces cerevisiae [Hinnen et al. (1978) Proc. Natl. Acad. Sci. USA 75:1929; Ito et al. (1983) J. Bacteriol. 755:163], Schizosaccharomyces pombe [Beach and Nurse (1981) Nature 300:706], and Yarrowia lipolytica [Davidow, et al. (1985) Curr. Genet. 70:380471 Gaillardin, et al. (1985) Curr. Genet. 70:49]. Methods of introducing exogenous DNA into yeast hosts are well-known in the art, and usually include either the transformation of spheroplasts or of intact yeast cells treated with alkali cations. Transformation procedures usually vary with the yeast species to be transformed. See eg. [Kurtz et al. (1986) Mol. Cell. Biol. 5:142; Kunze et al. (1985) J. Basic Microbiol. 25:141; Candida]; [Gleeson et al. (1986) J. Gen. Microbiol. 752:3459; Roggenkamp et al. (1986) Mol. Gen. Genet. 202:302; Hansenula]; [Das et al. (1984) J. Bacteriol. 755:1165; De Louvencourt et al. (1983) J. Bacteriol. 154:1165; Van den Berg et al. (1990) Bio/Technology 5:135; Kluyveromyces]; [Cregg et al. (1985) Mol. Cell. Biol. 5:3376; Kunze et al. (1985) J. Basic Microbiol. 25:141; US Patent Nos. 4,837,148 and 4,929,555; Pichia]; [Hinnen et al. (1978) Proc. Natl. Acad. Sci. USA 75;1929; Ito et al. (1983) J. Bacteriol. 755:163 Saccharomyces]; [Beach and Nurse (1981) Nature 300:706; Schizosaccharomyces]; [Davidow et al. (1985) Curr. Genet. 70:39; Gaillardin et al. (1985) Curr. Genet. 10:49; Yarrowia]. Antibodies As used herein, the term "antibody" refers to a polypeptide or group of polypeptides composed of at least one antibody combining site. An "antibody combining site" is the three-dimensional binding space with an internal surface shape and charge distribution complementary to the features of an epitope of an antigen, which allows a binding of the antibody with the antigen. "Antibody" includes, for example, vertebrate antibodies, hybrid antibodies, chimeric antibodies, humanised antibodies, altered antibodies, univalent .antibodies, Fab proteins, and single domain antibodies. Antibodies against the proteins of the invention are useful for affinity chromatography, immunoassays, and distinguishing/identifying Neisserial proteins. Antibodies to the proteins of the invention, both polyclonal and monoclonal, may be prepared by conventional methods. In general, the protein is first used to immunize a suitable animal, preferably a mouse, rat, rabbit or goat. Rabbits and goats are preferred for the preparation of polyclonal sera due to the volume of serum obtainable, and the availability of labeled anti-rabbit and anti-goat antibodies. Immunization is generally performed by mixing or emulsifying the protein in saline, preferably in an adjuvant such as Freund's complete adjuvant, and injecting the mixture or emulsion parenterally (generally subcutaneously or intramuscularly). A dose of 50-200 μg/injection is typically sufficient. Immunization is generally boosted 2-6 weeks later with one or more injections of the protein in saline, preferably using Freund's incomplete adjuvant. One may alternatively generate antibodies by in vitro immunization using methods known in the art, which for the purposes of this invention is considered equivalent to in vivo immunization. Polyclonal antisera is obtained by bleeding the immunized animal into a glass or plastic container, incubating the blood at 25°C for one hour, followed by incubating at 4°C for 2-18 hours. The serum is recovered by centrifugation (eg. l,000g for 10 minutes). About 20-50 ml per bleed may be obtained from rabbits. Monoclonal antibodies are prepared using the standard method of Kohler & Milstein [Nature (1975) 256:495-96], or a modification thereof. Typically, a mouse or rat is immunized as described above. However, rather than bleeding the animal to extract serum, the spleen (and optionally several large lymph nodes) is removed and dissociated into single cells. If desired, the spleen cells may be screened (after removal of nonspecifically adherent cells) by applying a cell suspension to a plate or well coated with the protein antigen. B-cells expressing membrane-bound i munoglobulin specific for the antigen bind to the plate, and are not rinsed away with the rest of the suspension. Resulting B-cells, or all dissociated spleen cells, are then induced to fuse with myeloma cells to form hybridomas, and are cultured in a selective medium (eg. hypoxanthine, aminopterin, thymidine medium, "HAT"). The resulting hybridomas are plated by limiting dilution, and are assayed for the production of antibodies which bind specifically to the immunizing antigen (and which do not bind to unrelated antigens). The selected MAb-secreting hybridomas are then cultured either in vitro (eg. in tissue culture bottles or hollow fiber reactors), or in vivo (as ascites in mice). If desired, the antibodies (whether polyclonal or monoclonal) may be labeled using conventional techniques. Suitable labels include fluorophores, chromophores, radioactive atoms (particularly 32 P and 125 I), electron-dense reagents, enzymes, and ligands having specific binding partners. Enzymes are typically detected by their activity. For example, horseradish peroxidase is usually detected by its ability to convert 3,3',5,5'-tetramethylbenzidine (TMB) to a blue pigment, quantifiable with a spectrophotometer. "Specific binding partner" refers to a protein capable of binding a ligand molecule with high specificity, as for example in the case of an antigen and a monoclonal antibody specific therefor. Other specific binding partners include biotin and avidin or streptavidin, IgG and protein A, and the numerous receptor-ligand couples known in the art. It should be understood that the above description is not meant to categorize the various labels into distinct classes, as the same label may serve in several different modes. For example, 125 I may serve as a radioactive label or as an electron-dense reagent. HRP may serve as enzyme or as antigen for a MAb. Further, one may combine various labels for desired effect. For ex.ample, MAbs .and avidin also require labels in the practice of this invention: thus, one might label a MAb with biotin, and detect its presence with avidin labeled with 125 I, or with an anti-biotin MAb labeled with HRP. Other permutations and possibilities will be readily apparent to those of ordinary skill in the art, and are considered as equivalents within the scope of the instant invention. Pharmaceutical Compositions Pharmaceutical compositions can comprise either polypeptides, antibodies, or nucleic acid of the invention. The pharmaceutical compositions will comprise a therapeutically effective amount of either polypeptides, antibodies, or polynucleotides of the claimed invention. The term "therapeutically effective amount" as used herein refers to an amount of a therapeutic agent to treat, ameliorate, or prevent a desired disease or condition, or to exhibit a detectable therapeutic or preventative effect. The effect can be detected by, for example, chemical markers or antigen levels. Therapeutic effects also include reduction in physical symptoms, such as decreased body temperature. The precise effective amount for a subject will depend upon the subject's size and health, the nature and extent of the condition, and the therapeutics or combination of therapeutics selected for administration. Thus, it is not useful to specify an exact effective amount in advance. However, the effective amount for a given situation can be determined by routine experimentation and is within the judgement of the clinician. For purposes of the present invention, an effective dose will be from about 0.01 mg/ kg to 50 mg kg or 0.05 mg/kg to about 10 mg/kg of the DNA constructs in the individual to which it is administered. A pharmaceutical composition can also contain a pharmaceutically acceptable carrier. The term "pharmaceutically acceptable carrier" refers to a carrier for administration of a therapeutic agent, such as antibodies or a polypeptide, genes, and other therapeutic agents. The term refers to any pharmaceutical carrier that does not itself induce the production of antibodies harmful to the individual receiving the composition, and which may be administered without undue toxicity. Suitable carriers may be large, slowly metabolized macromolecules such as proteins, polysaccharides, polylactic acids, polyglycolic acids, polymeric amino acids, amino acid copolymers, and inactive virus particles. Such carriers are well known to those of ordinary skill in the art. Pharmaceutically acceptable salts can be used therein, for example, mineral acid salts such as hydrochlorides, hydrobromides, phosphates, sulfates, and the like; and the salts of organic acids such as acetates, propionates, malonates, benzoates, and the like. A thorough discussion of pharmaceutically acceptable excipients is available in Remington's Pharmaceutical Sciences (Mack Pub. Co., N.J. 1991). Pharmaceutically acceptable carriers in therapeutic compositions may contain liquids such as water, saline, glycerol and ethanol. Additionally, auxiliary substances, such as wetting or emulsifying agents, pH buffering substances, .and the like, may be present in such vehicles. Typically, the therapeutic compositions are prepared as injectables, either as liquid solutions or suspensions; solid forms suitable for solution in, or suspension in, liquid vehicles prior to injection may also be prepared. Liposomes are included within the definition of a pharmaceutically acceptable carrier. Delivery Methods Once formulated, the compositions of the invention can be administered directly to the subject. The subjects to be treated can be animals; in particular, human subjects can be treated. Direct delivery of the compositions will generally be accomplished by injection, either subcutaneously, intraperitoneally, intravenously or intramuscularly or delivered to the interstitial space of a tissue. The compositions can also be administered into a lesion. Other modes of administration include oral and pulmonary administration, suppositories, and transdermal or transcutaneous applications (eg. see WO98/20734), needles, and gene guns or hyposprays. Dosage treatment may be a single dose schedule or a multiple dose schedule. Vaccines Vaccines according to the invention may either be prophylactic (ie. to prevent infection) or therapeutic (ie. to treat disease after infection). Such vaccines comprise immunising antigen(s), immunogen(s), polypeptide(s), protein(s) or nucleic acid, usually in combination with "pharmaceutically acceptable carriers," which include any carrier that does not itself induce the production of antibodies harmful to the individual receiving the composition. Suitable carriers are typically large, slowly metabolized macromolecules such as proteins, polysaccharides, polylactic acids, polyglycolic acids, polymeric amino acids, amino acid copolymers, lipid aggregates (such as oil droplets or liposomes), and inactive virus particles. Such carriers are well known to those of ordinary skill in the art. Additionally, these carriers may function as immunostimulating agents ("adjuvants"). Furthermore, the antigen or immunogen may be conjugated to a bacterial toxoid, such as a toxoid from diphtheria, tetanus, cholera, H. pylori, etc. pathogens. Preferred adjuvants to enhance effectiveness of the composition include, but are not limited to: (1) aluminum salts (alum), such as aluminum hydroxide, aluminum phosphate, aluminum sulfate, etc; (2) oil-in-water emulsion formulations (with or without other specific immunostimulating agents such as muramyl peptides (see below) or bacterial cell wall components), such as for example (a) MF59™ (WO 90/14837; Chapter 10 in Vaccine design: the subunit and adjuvant approach, eds. Powell & Newm.an, Plenum Press 1995), containing 5% Squalene, 0.5% Tween 80, and 0.5% Span 85 (optionally containing various amounts of MTP-PE (see below), although not required) formulated into submicron particles using a microfluidizer such as Model HOY microfiuidizer (Microfluidics, Newton, MA), (b) SAF, containing 10% Squalane, 0.4% Tween 80, 5% pluronic- blocked polymer L121, and thr-MDP (see below) either microfluidized into a submicron emulsion or vortexed to generate a larger particle size emulsion, and (c) Ribi™ adjuvant system (RAS), (Ribi Immunochem, Hamilton, MT) containing 2% Squalene, 0.2% Tween 80, and one or more bacterial cell wall components from the group consisting of monophosphorylipid A (MPL), trehalose dimycolate (TDM), and cell wall skeleton (CWS), preferably MPL + CWS (Detox™); (3) saponin adjuvants, such as Stimulon™ (Cambridge Bioscience, Worcester, MA) may be used or particles generated therefrom such as ISCOMs (immunostimulating complexes); (4) Complete Freund's Adjuvant (CFA) and Incomplete Freund's Adjuvant (IF A); (5) cytokines, such as interleukins (eg. IL-1, IL-2, IL-4, IL-5, IL-6, IL-7, IL-12, etc.), interferons (eg. gamma interferon), macrophage colony stimulating factor (M-CSF), tumor necrosis factor (TNF), etc; and (6) other substances that act as immunostimulating agents to enhance the effectiveness of the composition. Alum and MF59™ are preferred. As mentioned above, muramyl peptides include, but are not limited to, N-acetyl-muramyl-L- threonyl-D-isoglutamine (thr-MDP), N-acetyl-normuramyl-L-alanyl-D-isoglutamine (nor-MDP), N-acetylmuramyl-L-alanyl-D-isoglutaminyl-L-alanine-2-( 1 '-2'-dipalmitoyl-5«-glycero-3 - hydroxyphosphoryloxy)-ethylamine (MTP-PE), etc. The immunogenic compositions (eg. the immunising antigen/immunogen/polypeptide/protein nucleic acid, pharmaceutically acceptable carrier, and adjuvant) typically will contain diluents, such as water, saline, glycerol, ethanol, etc. Additionally, auxiliary substances, such as wetting or emulsifying agents, pH buffering substances, and the like, may be present in such vehicles. Typically, the immunogenic compositions are prepared as injectables, either as liquid solutions or suspensions; solid forms suitable for solution in, or suspension in, liquid vehicles prior to injection may also be prepared. The preparation also may be emulsified or encapsulated in liposomes for enhanced adjuvant effect, as discussed above under pharmaceutically acceptable carriers. Immunogenic compositions used as vaccines comprise an immunologically effective amount of the antigenic or immunogenic polypeptides, as well as any other of the above-mentioned components, as needed. By "immunologically effective amount", it is meant that the administration of that amount to an individual, either in a single dose or as part of a series, is effective for treatment or prevention. This amount varies depending upon the health and physical condition of the individual to be treated, the taxonomic group of individual to be treated (eg. nonhuman primate, primate, etc.), the capacity of the individual's immune system to synthesize antibodies, the degree of protection desired, the formulation of the vaccine, the treating doctor's assessment of the medical situation, and other relevant factors. It is expected that the amount will fall in a relatively broad range that can be determined through routine trials. The immunogenic compositions are conventionally administered parenterally, eg. by injection, either subcutaneously, intramuscularly, or transdermally/transcutaneously (eg. WO98/20734). Additional formulations suitable for other modes of administration include oral and pulmonary formulations, suppositories, and transdermal applications. Dosage treatment may be a single dose schedule or a multiple dose schedule. The vaccine may be administered in conjunction with other immunoregulatory agents. As an alternative to protein-based vaccines, DNA vaccination may be employed [eg. Robinson & Torres (1997) Seminars in Immunology 9:271-283; Donnelly et al. (1997) Annu Rev Immunol 15:617-648; see later herein]. Gene Delivery Vehicles Gene therapy vehicles for delivery of constructs including a coding sequence of a therapeutic of the invention, to be delivered to the mammal for expression in the mammal, can be administered either locally or systemically. These constructs can utilize viral or non- viral vector approaches in in vivo or ex vivo modality. Expression of such coding sequence can be induced using endogenous mammalian or heterologous promoters. Expression of the coding sequence in vivo can be either constitutive or regulated. The invention includes gene delivery vehicles capable of expressing the contemplated nucleic acid sequences. The gene delivery vehicle is preferably a viral vector and, more preferably, a retroviral, adenoviral, adeno-associated viral (AAV), herpes viral, or alphavirus vector. The viral vector can also be an astrovirus, coronavirus, orthomyxovirus, papovavirus, paramyxovirus, parvovirus, picornavirus, poxvirus, or togavirus viral vector. See generally, Jolly (1994) Cancer Gene Therapy 1:51-64; Kimura (1994) Human Gene Therapy 5:845-852; Connelly (1995) Human Gene Therapy 6:185-193; and Kaplitt (1994) Nature Genetics 6:148-153. Retroviral vectors .are well known in the art and we contemplate that any retroviral gene therapy vector is employable in the invention, including B, C and D type retroviruses, xenotropic retroviruses (for example, NZB-X1, NZB-X2 and NZB9-1 (see O'Neill (1985) J. Virol. 53:160) polytropic retroviruses eg. MCF and MCF-MLV (see Kelly (1983) J. Virol. 45:291), spumaviruses and lentiviruses. See RNA Tumor Viruses, Second Edition, Cold Spring Harbor Laboratory, 1985. Portions of the retroviral gene therapy vector may be derived from different retroviruses. For example, retro vector LTRs may be derived from a Murine Sarcoma Virus, a tRNA binding site from a Rous Sarcoma Virus, a packaging signal from a Murine Leukemia Virus, and an origin of second strand synthesis from an Avian Leukosis Virus. These recombinant retroviral vectors may be used to generate transduction competent retroviral vector particles by introducing them into appropriate packaging cell lines (see US patent 5,591,624). Retrovirus vectors can be constructed for site-specific integration into host cell DNA by incorporation of a chimeric integrase enzyme into the retroviral particle (see WO96/37626). It is preferable that the recombinant viral vector is a replication defective recombinant virus. Packaging cell lines suitable for use with the above-described retrovirus vectors are well known in the art, are readily prepared (see WO95/30763 and WO92/05266), and can be used to create producer cell lines (also termed vector cell lines or "VCLs") for the production of recombinant vector particles. Preferably, the packaging cell lines are made from human parent cells (eg. HT1080 cells) or mink parent cell lines, which eliminates inactivation in human serum. Preferred retroviruses for the construction of retroviral gene therapy vectors include Avian Leukosis Virus, Bovine Leukemia, Virus, Murine Leukemia Virus, Mink-Cell Focus-Inducing Virus, Murine Sarcoma Virus, Reticuloendothehosis Virus and Rous Sarcoma Virus. Particularly preferred Murine Leukemia Viruses include 4070A and 1504A (Hartley and Rowe (1976) J Virol 19:19-25), Abelson (ATCC No. VR-999), Friend (ATCC No. VR-245), Graffi, Gross (ATCC Nol VR-590), Kirsten, Harvey Sarcoma Virus and Rauscher (ATCC No. VR-998) and Moloney Murine Leukemia Virus (ATCC No. VR-190). Such retroviruses may be obtained from depositories or collections such as the American Type Culture Collection ("ATCC") in Rockville, Maryland or isolated from known sources using commonly available techniques. Exemplary known retroviral gene therapy vectors employable in this invention include those described in patent applications GB2200651, EP0415731, EP0345242, EP0334301, WO89/02468; WO89/05349, WO89/09271, WO90/02806, WO90/07936, WO94/03622, WO93/25698, WO93/25234, WO93/11230, WO93/10218, WO91/02805, WO91/02825, WO95/07994, US 5,219,740, US 4,405,712, US 4,861,719, US 4,980,289, US 4,777,127, US 5,591,624. See also Vile (1993) Cancer Res 53:3860-3864; Vile (1993) Cancer Res 53:962-967; Ram (1993) Cancer Res 53 (1993) 83-88; Takamiya (1992) J Neurosci Res 33:493-503; Baba (1993) J Neurosurg 79:729-735; Mann (1983) Cell 33:153; Cane (1984) Proc Natl Acad Sci 81:6349; .and Miller (1990) Human Gene Therapy 1. Human adenoviral gene therapy vectors are also known in the art and employable in this invention. See, for example, Berkner (1988) Biotechniques 6:616 and Rosenfeld (1991) Science 252:431, and WO93/07283, WO93/06223, and WO93/07282. Exemplary known adenoviral gene therapy vectors employable in this invention include those described in the above referenced documents and in WO94/12649, WO93/03769, WO93/19191, WO94/28938, WO95/11984, WO95/00655, WO95/27071, WO95/29993, WO95/34671, WO96/05320, WO94/08026, WO94/11506, WO93/06223, WO94/24299, WO95/14102, WO95/24297, WO95/02697, WO94/28152, WO94/24299, WO95/09241, WO95/25807, WO95/05835, WO94/18922 and WO95/09654. Alternatively, administration of DNA linked to killed adenovirus as described in Curiel (1992) Hum. Gene Ther. 3:147-154 may be employed. The gene delivery vehicles of the invention also include adenovirus associated virus (AAV) vectors. Leading and preferred examples of such vectors for use in this invention are the AAV-2 based vectors disclosed in Srivastava, WO93/09239. Most preferred AAV vectors comprise the two AAV inverted terminal repeats in which the native D-sequences are modified by substitution of nucleotides, such that at least 5 native nucleotides and up to 18 native nucleotides, preferably at least 10 native nucleotides up to 18 native nucleotides, most preferably 10 native nucleotides are retained and the remaining nucleotides of the D-sequence are deleted or replaced with non-native nucleotides. The native D-sequences of the AAV inverted terminal repeats are sequences of 20 consecutive nucleotides in each AAV inverted terminal repeat (ie. there is one sequence at each end) which are not involved in HP formation. The non-native replacement nucleotide may be any nucleotide other than the nucleotide found in the native D-sequence in the same position. Other employable exemplary AAV vectors are pWP-19, pWN-1, both of which are disclosed in Nahreini (1993) Gene 124:257-262. Another example of such an AAV vector is psub201 (see Samulski (1987) J Virol. 61:3096). Another exemplary AAV vector is the Double-D ITR vector. Construction of the Double-D ITR vector is disclosed in US Patent 5,478,745. Still other vectors are those disclosed in Carter US Patent 4,797,368 and MuzyczkaUS Patent 5,139,941, Chartejee US Patent 5,474,935, and Kotin WO94/288157. Yet a further example of an AAV vector employable in this invention is SSV9AFABTKneo, which contains the AFP enhancer and albumin promoter and directs expression predominantly in the liver. Its structure and construction are disclosed in Su (1996) Human Gene Therapy 7:463-470. Additional AAV gene therapy vectors are described in US 5,354,678, US 5,173,414, US 5,139,941, and US 5,252,479. The gene therapy vectors of the invention also include herpes vectors. Leading and preferred examples are herpes simplex virus vectors containing a sequence encoding a thymidine kinase polypeptide such as those disclosed in US 5,288,641 and EP0176170 (Roizman). Additional exemplary herpes simplex virus vectors include HFEM/ICP6-LacZ disclosed in WO95/04139 (Wistar Institute), pHSVlac described in Geller (1988) Science 241:1667-1669 and in WO90/09441 and WO92/07945, HSV Us3::pgC-lacZ described in Fink (1992) Human Gene Therapy 3:11-19 and HSV 7134, 2 RH 105 and GAL4 described in EP 0453242 (Breakefield), and those deposited with the ATCC as accession numbers ATCC VR-977 and ATCC VR-260. Also contemplated are alpha virus gene therapy vectors that can be employed in this invention. Preferred alpha virus vectors are Sindbis viruses vectors. Togaviruses, Semliki Forest virus (ATCC VR-67; ATCC VR-1247), Middleberg virus (ATCC VR-370), Ross River virus (ATCC VR-373; ATCC VR-1246), Venezuelan equine encephalitis virus (ATCC VR923; ATCC VR-1250; ATCC VR-1249; ATCC VR-532), and those described in US patents 5,091,309, 5,217,879, and WO92/10578. More particularly, those alpha virus vectors described in US Serial No. 08/405,627, filed March 15, 1995,WO94/21792, WO92/10578, WO95/07994, US 5,091,309 and US 5,217,879 are employable. Such alpha viruses may be obtained from depositories or collections such as the ATCC in Rockville, Maryland or isolated from known sources using commonly available techniques. Preferably, alphavirus vectors with reduced cytotoxicity are used (see USSN 08/679640). DNA vector systems such as eukarytic layered expression systems are also useful for expressing the nucleic acids of the invention. See WO95/07994 for a detailed description of eukaryotic layered expression systems. Preferably, the eukaryotic layered expression systems of the invention are derived from alphavirus vectors and most preferably from Sindbis viral vectors. Other viral vectors suitable for use in the present invention include those derived from poliovirus, for example ATCC VR-58 and those described in Evans, Nature 339 (1989) 385 and Sabin (1973) J. Biol. Standardization 1:115; rhinovirus, for example ATCC VR-1110 and those described in Arnold (1990) J Cell Biochem L401; pox viruses such as canary pox virus or vaccinia virus, for example ATCC VR-111 and ATCC VR-2010 and those described in Fisher-Hoch (1989) Proc Natl Acad Sci 86:317; Flexner (1989) Ann NY Acad Sci 569:86, Flexner (1990) Vaccine 8:17; in US 4,603,112 and US 4,769,330 and WO89/01973; SV40 virus, for example ATCC VR-305 and those described in Mulligan (1979) Nαtwre 277:108 and Madzak (1992) J Gen Virol 73:1533; influenza virus, for example ATCC VR-797 and recombinant influenza viruses made employing reverse genetics techniques as described in US 5,166,057 and in Enami (1990) Proc Natl Acad Sci 87:3802-3805; Enami & Palese (1991) J Virol 65:2711-2713 and Luytjes (1989) Cell 59:110, (see also McMichael (1983) NEJ Med 309:13, and Yap (1978) Nαtwre 273:238 and Nαtwre (1979) 277:108); human immunodeficiency virus as described in EP-0386882 and in Buchschacher (1992) J Virol. 66:2731; measles virus, for example ATCC VR-67 and VR-1247 and those described in EP-0440219; Aura virus, for example ATCC VR-368; Bebaru virus, for example ATCC VR-600 and ATCC VR-1240; Cabassou virus, for example ATCC VR-922; Chikungunya virus, for example ATCC VR-64 and ATCC VR-1241; Fort Morgan Virus, for example ATCC VR-924; Getah virus, for example ATCC VR-369 and ATCC VR-1243; Kyzylagach virus, for example ATCC VR-927; Mayaro virus, for example ATCC VR-66; Mucambo virus, for example ATCC VR-580 and ATCC VR-1244; Νdumu virus, for example ATCC VR-371; Pixuna virus, for example ATCC VR-372 and ATCC VR-1245; Tonate virus, for example ATCC VR-925; Triniti virus, for example ATCC VR-469; Una virus, for example ATCC VR-374; Whataroa virus, for example ATCC VR-926; Y-62-33 virus, for example ATCC VR-375; O'Νyong virus, Eastern encephalitis virus, for example ATCC VR-65 and ATCC VR-1242; Western encephalitis virus, for example ATCC VR-70, ATCC VR-1251, ATCC VR-622 and ATCC VR-1252; and coronavirus, for example ATCC VR-740 and those described in Hamre (1966) Proc Soc Exp Biol Med 121 : 190. Delivery of the compositions of this invention into cells is not limited to the above mentioned viral vectors. Other delivery methods and media may be employed such as, for example, nucleic acid expression vectors, polycationic condensed DNA linked or unlinked to killed adenovirus alone, for example see US Serial No. 08/366,787, filed December 30, 1994 and Curiel (1992) Hum Gene Ther 3:147-154 ligand linked DNA, for example see Wu (1989) J Biol Chem 264:16985-16987, eucaryotic cell delivery vehicles cells, for example see US Serial No.08/240,030, filed May 9, 1994, and US Serial No. 08/404,796, deposition of photopolymerized hydrogel materials, hand-held gene transfer particle gun, as described in US Patent 5,149,655, ionizing radiation as described in US5,206,152 and in WO92/11033, nucleic charge neutralization or fusion with cell membranes. Additional approaches are described in Philip (1994) Mol Cell Biol 14:2411-2418 and in Woffendin (1994) Proc Natl Acad Sci 91:1581-1585. Particle mediated gene transfer may be employed, for example see US Serial No. 60/023,867. Briefly, the sequence can be inserted into conventional vectors that contain conventional control sequences for high level expression, and then incubated with synthetic gene transfer molecules such as polymeric DNA-binding cations like polylysine, protamine, and albumin, linked to cell targeting ligands such as asialoorosomucoid, as described in Wu 8c Wu (1987) J. Biol. Chem. 262:.4429-4432, insulin as described in Hucked (1990) Biochem Pharmacol 40:253-263, galactose as described in Plank (1992) Bioconjugate Chem 3:533-539, lactose or transferrin. Naked DNA may also be employed. Exemplary naked DNA introduction methods are described in WO 90/11092 and US 5,580,859. Uptake efficiency may be improved using biodegradable latex beads. DNA coated latex beads are efficiently transported into cells after endocytosis initiation by the beads. The method may be improved further by treatment of the beads to increase hydrophobicity and thereby facilitate disruption of the endosome and release of the DNA into the cytoplasm. Liposomes that can act as gene delivery vehicles are described in US 5,422,120, WO95/13796, WO94/23697, WO91/14445 and EP-524,968. As described in USSN. 60/023,867, on non-viral delivery, the nucleic acid sequences encoding a polypeptide can be inserted into conventional vectors that contain conventional control sequences for high level expression, and then be incubated with synthetic gene transfer molecules such as polymeric DNA-binding cations like polylysine, protamine, and albumin, linked to cell targeting ligands such as asialoorosomucoid, insulin, galactose, lactose, or transferrin. Other delivery systems include the use of liposomes to encapsulate DNA comprising the gene under the control of a variety of tissue-specific or ubiquitously-active promoters. Further non-viral delivery suitable for use includes mechanical delivery systems such as the approach described in Woffendin et al (1994) Proc. Natl. Acad. Sci. USA 91(24): 11581-11585. Moreover, the coding sequence and the product of expression of such can be delivered through deposition of photopolymerized hydrogel materials. Other conventional methods for gene delivery that can be used for delivery of the coding sequence include, for example, use of hand-held gene transfer particle gun, as described in US 5,149,655; use of ionizing radiation for activating transferred gene, as described in US 5,206, 152 and WO92/11033 Exemplary liposome and polycationic gene delivery vehicles are those described in US 5,422,120 and 4,762,915; inWO 95/13796; WO94/23697; and WO91/14445; in EP-0524968; and in Stryer, Biochemistry, pages 236-240 (1975) W.H. Freeman, San Francisco; Szoka (1980) Biochem BiophysActa 600:1; Bayer (1979) Biochem Biophys Acta 550:464; Rivnay (1987) Meth Enzymol 149:119; Wang (1987) Proc Natl Acad Sci 84:7851; Plant (1989) Anal Biochem 176:420. A polynucleotide composition can comprises therapeutically effective amount of a gene therapy vehicle, as the term is defined above. For purposes of the present invention, an effective dose will be from about 0.01 mg/ kg to 50 mg/kg or 0.05 mg/kg to about 10 mg/kg of the DNA constructs in the individual to which it is administered. Delivery Methods Once formulated, the polynucleotide compositions of the invention can be administered (1) directly to the subject; (2) delivered ex vivo, to cells derived from the subject; or (3) in vitro for expression of recombinant proteins. The subjects to be treated can be mammals or birds. Also, human subjects can be treated. Direct delivery of the compositions will generally be accomplished by injection, either subcutaneously, intraperitoneally, intravenously or intramuscularly or delivered to the interstitial space of a tissue. The compositions can also be administered into a lesion. Other modes of administration include oral and pulmonary administration, suppositories, and transdermal or transcutaneous applications (eg. see WO98/20734), needles, and gene guns or hyposprays. Dosage treatment may be a single dose schedule or a multiple dose schedule. Methods for the ex vivo delivery and reimplantation of transformed cells into a subject are known in the art and described in eg. WO93/14778. Examples of cells useful in ex vivo applications include, for example, stem cells, particularly hematopoetic, lymph cells, macrophages, dendritic cells, or tumor cells. Generally, delivery of nucleic acids for both ex vivo and in vitro applications can be accomplished by the following procedures, for example, dextran-mediated transfection, calcium phosphate precipitation, polybrene mediated transfection, protoplast fusion, electroporation, encapsulation of the polynucleotide(s) in liposomes, and direct microinjection of the DNA into nuclei, all well known in the art. Polynucleotide and yolvpeptide pharmaceutical compositions In addition to the pharmaceutically acceptable carriers and salts described above, the following additional agents can be used with polynucleotide and/or polypeptide compositions. A.Polvpeptides One example are polypeptides which include, without limitation: asioloorosomucoid (ASOR); transferrin; asialoglycoproteins; antibodies; antibody fragments; ferritin; interleukins; interferons, granulocyte, macrophage colony stimulating factor (GM-CSF), granulocyte colony stimulating factor (G-CSF), macrophage colony stimulating factor (M-CSF), stem cell factor and erythropoietin. Viral antigens, such as envelope proteins, can also be used. Also, proteins from other invasive organisms, such as the 17 amino acid peptide from the circumsporozoite protein of plasmodium falciparum known as RII. B .Hormones, Vitamins, etc. Other groups that can be included are, for example: hormones, steroids, androgens, estrogens, thyroid hormone, or vitamins, folic acid. C.Polyalkylenes. Polysaccharides. etc. Also, polyalkylene glycol can be included with the desired polynucleotides/polypeptides. In a preferred embodiment, the polyalkylene glycol is polyethlylene glycol. In addition, mono-, di-, or polysaccarides can be included. In a preferred embodiment of this aspect, the polysaccharide is dextran or DEAE-dextran. Also, chitosan and poly(lactide-co-glycolide) D.Lipids. and Liposomes The desired polynucleoti.de/polypeptide can also be encapsulated in lipids or packaged in liposomes prior to delivery to the subject or to cells derived therefrom. Lipid encapsulation is generally accomplished using liposomes which are able to stably bind or entrap and retain nucleic acid. The ratio of condensed polynucleotide to lipid preparation can vary but will generally be around 1:1 (mg DNA:micromoles lipid), or more of hpid. For a review of the use of liposomes as carriers for delivery of nucleic acids, see, Hug and Sleight (1991) Biochim. Biophys. Acta. 1097:1-17; Straubinger (1983) Meth. Enzymol. 101:512-527. Liposomal preparations for use in the present invention include cationic (positively charged), anionic (negatively charged) and neutral preparations. Cationic liposomes have been shown to mediate intracellular delivery of plasmid DNA (Feigner (1987) Proc. Natl. Acad. Sci. USA 84:7413-7416); mRNA (Malone (1989) Proc. Natl. Acad. Sci. USA 86:6077-6081); and purified transcription factors (Debs (1990) J. Biol. Chem. 265:10189-10192), in functional form. Cationic liposomes are readily available. For example, N[l-2,3-dioleyloxy)propyl]-N,N,N-triethylammonium (DOTMA) liposomes are available under the trademark Lipofectin, from GLBCO BRL, Grand Island, NY. (See, also, Feigner supra). Other commercially available liposomes include transfectace (DDAB/DOPE) and DOTAP/DOPE (Boerhinger). Other cationic liposomes can be prepared from readily available materials using techniques well known in the art. See, eg. Szoka (1978) Proc. Natl. Acad. Sci. USA 75:4194-4198; WO90/11092 for a description of the synthesis of DOTAP (l,2-bis(oleoyloxy)-3-(trimethylammonio)propane) liposomes. Similarly, anionic and neutral liposomes are readily available, such as from Avanti Polar Lipids (Birmingham, AL), or can be easily prepared using readily available materials. Such materials include phosphatidyl choline, cholesterol, phosphatidyl ethanolamine, dioleoylphosphatidyl choline (DOPC), dioleoylphosphatidyl glycerol (DOPG), dioleoylphoshatidyl ethanolamine (DOPE), among others. These materials can also be mixed with the DOTMA and DOTAP starting materials in appropriate ratios. Methods for making liposomes using these materials are well known in the art. The liposomes can comprise multilammelar vesicles (MLVs), small unilamellar vesicles (SUVs), or large unilamellar vesicles (LUVs). The various liposome-nucleic acid complexes are prepared using methods known in the art. See eg. Straubinger (1983) Meth. Immunol. 101 :512-527; Szoka (1978) Proc. Nαtl. Acαd. Sci. USA 75:4194-4198; Papahadjopoulos (1975) Biochim. Biophys. Actα 394:483; Wilson (1979) Cell 17:77); Deamer & Bangham (1976) Biochim. Biophys. Actα 443:629; Ostro (1977) Biochem. Biophys. Res. Commun. 76:836; Fraley (1979) Proc. Nαtl. Acαd. Sci. USA 76:3348); Enoch & Strittmatter (1979) Proc. Nαtl. Acαd. Sci. USA 76:145; Fraley (1980) J. Biol. Chem. (1980) 255:10431; Szoka & Papahadjopoulos (1978) Proc. Nαtl. Acαd. Sci. USA 75:145; and Schaefer-Ridder (1982) Science 215:166. E.Lipoproteins In addition, lipoproteins can be included with the polynucleotide/polypeptide to be delivered. Examples of lipoproteins to be utilized include: chylomicrons, HDL, IDL, LDL, and VLDL. Mutants, fragments, or fusions of these proteins can also be used. Also, modifications of naturally occurring lipoproteins can be used, such as acetylated LDL. These lipoproteins can target the delivery of polynucleotides to cells expressing lipoprotein receptors. Preferably, if lipoproteins are including with the polynucleotide to be delivered, no other targeting ligand is included in the composition. Naturally occurring lipoproteins comprise a lipid and a protein portion. The protein portion are known as apoproteins. At the present, apoproteins A, B, C, D, and E have been isolated and identified. At least two of these contain several proteins, designated by Roman numerals, Al, All, ATV; Cl, CII, CIII. A lipoprotein can comprise more than one apoprotein. For example, naturally occurring chylomicrons comprises of A, B, C, and E, over time these lipoproteins lose A and acquire C and E apoproteins. VLDL comprises A, B, C, and E apoproteins, LDL comprises apoprotein B; and HDL comprises apoproteins A, C, and E. The amino acid of these apoproteins are known and are described in, for example, Breslow (1985) Annu Rev. Biochem 54:699; Law (1986) Adv. Exp Med. Biol. 151 :162; Chen (1986) J Biol Chem 261:12918; Kane (1980) Proc Natl Acad Sci USA 77:2465; and Utermann (1984) Hum Genet 65:232. Lipoproteins contain a variety of lipids including, triglycerides, cholesterol (free and esters), and phopholipids. The composition of the lipids varies in naturally occurring lipoproteins. For example, chylomicrons comprise mainly triglycerides. A more detailed description of the lipid content of naturally occurring lipoproteins can be found, for example, in Meth. Enzymol. 128 (1986). The composition of the lipids are chosen to aid in conformation of the apoprotein for receptor binding activity. The composition of lipids can also be chosen to facilitate hydrophobic interaction and association with the polynucleotide binding molecule. Naturally occurring lipoproteins can be isolated from serum by ultracentrifugation, for instance. Such methods are described in Meth. Enzymol. (supra); Pitas (1980) J. Biochem. 255:5454-5460 and Mahey (1979) J Clin. Invest 64:743-750. Lipoproteins can also be produced by in vitro or recombinant methods by expression of the apoprotein genes in a desired host cell. See, for example, Atkinson (1986) Annu Rev Biophys Chem 15 :403 and Radding (1958) Biochim Biophys Acta 30: 443. Lipoproteins can also be purchased from commercial suppliers, such as Biomedical Techniologies, Inc., Stoughton, Massachusetts, USA. Further description of lipoproteins can be found in Zuckermann et al. PCT/US97/14465. F.Polvcationic Agents Polycationic agents can be included, with or without lipoprotein, in a composition with the desired polynucleotide/polypeptide to be delivered. Polycationic agents, typically, exhibit a net positive charge at physiological relevant pH and are capable of neutralizing the electrical charge of nucleic acids to facilitate delivery to a desired location. These agents have both in vitro, ex vivo, and in vivo applications. Polycatiomc agents can be used to deliver nucleic acids to a living subject either intramuscularly, subcutaneously, etc. The following are examples of useful polypeptides as polycationic agents: polylysine, polyarginine, polyorrώhine, and protamine. Other examples include histones, protamines, human serum albumin, DNA binding proteins, non-histone chromosomal proteins, coat proteins from DNA viruses, such as (XI 74, transcriptional factors also contain domains that bind DNA and therefore may be useful as nucleic aid condensing agents. Briefly, transcriptional factors such as C/CEBP, c-jun, c-fos, AP-1, AP-2, AP-3, CPF, Prot-1, Sp-1, Oct-1, Oct-2, CREP, and TF1TD contain basic domains that bind DNA sequences. Organic polycationic agents include: spermine, spermidine, and purtrescine. The dimensions and of the physical properties of a polycationic agent can be extrapolated from the list above, to construct other polypeptide polycationic agents or to produce synthetic polycatiomc agents. Synthetic polycationic agents which are useful include, for example, DEAE-dextran, polybrene. Lipofectin™, and lipofectAMLNE™ are monomers that form polycationic complexes when combined with polynucleotides/polypeptides. Immunodiasnostic Assays Neisserial antigens of the invention can be used in immunoassays to detect antibody levels (or, conversely, anti-Neisserial antibodies can be used to detect antigen levels). Immunoassays based on well defined, recombinant antigens can be developed to replace invasive diagnostics methods. Antibodies to Neisserial proteins within biological samples, including for example, blood or serum samples, can be detected. Design of the immunoassays is subject to a great deal of variation, and a variety of these are known in the art. Protocols for the immunoassay may be based, for example, upon competition, or direct reaction, or sandwich type assays. Protocols may also, for example, use solid supports, or may be by immunoprecipitation. Most assays involve the use of labeled antibody or polypeptide; the labels may be, for example, fluorescent, chemiluminescent, radioactive, or dye molecules. Assays which amplify the signals from the probe are also known; examples of which are assays which utilize biotin and avidin, and enzyme-labeled and mediated immunoassays, such as ELISA assays. Kits suitable for immunodiagnosis and containing the appropriate labeled reagents are constructed by packaging the appropriate materials, including the compositions of the invention, in suitable containers, along with the remaining reagents and materials (for example, suitable buffers, salt solutions, etc.) required for the conduct of the assay, as well as suitable set of assay instructions. Nucleic Acid Hybridisation "Hybridization" refers to the association of two nucleic acid sequences to one another by hydrogen bonding. Typically, one sequence will be fixed to a solid support and the other will be free in solution. Then, the two sequences will be placed in contact with one another under conditions that favor hydrogen bonding. Factors that affect this bonding include: the type and volume of solvent; reaction temperature; time of hybridization; agitation; agents to block the non-specific attachment of the liquid phase sequence to the solid support (Denhardt's reagent or BLOTTO); concentration of the sequences; use of compounds to increase the rate of association of sequences (dextran sulfate or polyethylene glycol); and the stringency of the washing conditions following hybridization. See Sambrook et al. [supra] Volume 2, chapter 9, pages 9.47 to 9.57. "Stringency" refers to conditions in a hybridization reaction that favor association of very similar sequences over sequences that differ. For example, the combination of temperature and salt concentration should be chosen that is approximately 120 to 200°C below the calculated Tm of the hybrid under study. The temperature and salt conditions can often be determined empirically in preliminary experiments in which samples of genomic DNA immobilized on filters are hybridized to the sequence of interest and then washed under conditions of different stringencies. See Sambrook et al. at page 9.50. Variables to consider when performing, for example, a Southern blot are (1) the complexity of the DNA being blotted and (2) the homology between the probe and the sequences being detected. The total amount of the fragment(s) to be studied can vary a magnitude of 10, from 0.1 to lμg for a plasmid or phage digest to IO "9 to 10 "8 g for a single copy gene in a highly complex eukaryotic genome. For lower complexity polynucleotides, substantially shorter blotting, hybridization, and exposure times, a smaller amount of starting polynucleotides, and lower specific activity of probes can be used. For example, a single-copy yeast gene can be detected with an exposure time of only 1 hour starting with 1 μg of yeast DNA, blotting for two hours, and hybridizing for 4-8 hours with a probe of IO 8 cpm/μg. For a single-copy mammalian gene a conservative approach would start with 10 μg of DNA, blot overnight, .and hybridize overnight in the presence of 10% dextran sulfate using a probe of greater than 10 s cpm/μg, resulting in an exposure time of -24 hours. Several factors can affect the melting temperature (Tm) of a DNA-DNA hybrid between the probe and the fragment of interest, and consequently, the appropriate conditions for hybridization and washing. In many cases the probe is not 100% homologous to the fragment. Other commonly encountered variables include the length and total G+C content of the hybridizing sequences and the ionic strength and formamide content of the hybridization buffer. The effects of all of these factors can be approximated by a single equation: Tm= 81 + 16.6(log 10 Ci) + 0.4[%(G + C)]-0.6(%formamide) - 600/n-1.5(%mismatch). where Ci is the salt concentration (monovalent ions) and n is the length of the hybrid in b.ase pairs (slightly modified from Meinkoth & Wahl (1984) Anal. Biochem. 138: 267-284). In designing a hybridization experiment, some factors affecting nucleic acid hybridization can be conveniently altered. The temperature of the hybridization and washes and the salt concentration during the washes are the simplest to adjust. As the temperature of the hybridization increases (ie. stringency), it becomes less likely for hybridization to occur between str.ands that are nonhomologous, and as a result, background decreases. If the radiolabeled probe is not completely homologous with the immobilized fragment (as is frequently the case in gene family and interspecies hybridization experiments), the hybridization temperature must be reduced, and background will increase. The temperature of the washes affects the intensity of the hybridizing band and the degree of background in a similar manner. The stringency of the washes is also increased with decreasing salt concentrations. In general, convenient hybridization temperatures in the presence of 50% formamide are 42°C for a probe with is 95% to 100% homologous to the target fragment, 37°C for 90% to 95% homology, and 32°C for 85% to 90% homology. For lower homologies, formamide content should be lowered and temperature adjusted accordingly, using the equation above. If the homology between the probe and the target fragment are not known, the simplest approach is to start with both hybridization and wash conditions which are nonstringent. If non-specific bands or high background are observed after autoradiography, the filter can be washed at high stringency .and reexposed. If the time required for exposure makes this approach impractical, several hybridization and/or washing stringencies should be tested in parallel. Nucleic Acid Probe Assays Methods such as PCR, branched DNA probe assays, or blotting techniques utilizing nucleic acid probes according to the invention can determine the presence of cDNA or mRNA. A probe is said to "hybridize" with a sequence of the invention if it can form a duplex or double stranded complex, which is stable enough to be detected. The nucleic acid probes will hybridize to the Neisserial nucleotide sequences of the invention (including both sense and antisense strands). Though many different nucleotide sequences will encode the amino acid sequence, the native Neisserial sequence is preferred because it is the actual sequence present in cells. mRNA represents a coding sequence and so a probe should be complementary to the coding sequence; single-stranded cDNA is complementary to mRNA, and so a cDNA probe should be complementary to the non-coding sequence. The probe sequence need not be identical to the Neisserial sequence (or its complement) — some variation in the sequence and length can lead to increased assay sensitivity if the nucleic acid probe can form a duplex with target nucleotides, which can be detected. Also, the nucleic acid probe can include additional nucleotides to stabilize the formed duplex. Additional Neisserial sequence may also be helpful as a label to detect the formed duplex. For example, a non-complementary nucleotide sequence may be attached to the 5' end of the probe, with the remainder of the probe sequence being complementary to a Neisserial sequence. Alternatively, non-complementary bases or longer sequences can be interspersed into the probe, provided that the probe sequence has sufficient complementarity with the a Neisserial sequence in order to hybridize therewith and thereby form a duplex which can be detected. The exact length and sequence of the probe will depend on the hybridization conditions, such as temperature, salt condition and the like. For example, for diagnostic applications, depending on the complexity of the analyte sequence, the nucleic acid probe typically contains at least 10-20 nucleotides, preferably 15-25, and more preferably at least 30 nucleotides, although it may be shorter than this. Short primers generally require cooler temperatures to form sufficiently stable hybrid complexes with the template. Probes may be produced by synthetic procedures, such as the triester method of Matteucci et al. [J. Am. Chem. Soc. (1981) 103:3185], or according to Urdea et al. [Proc. Natl. Acad. Sci. USA (1983) 80: 7461], or using commercially available automated oligonucleotide synthesizers. The chemical nature of the probe can be selected according to preference. For certain applications, DNA or RNA are appropriate. For other applications, modifications may be incorporated eg. backbone modifications, such as phosphorothioates or methylphosphonates, can be used to increase in vivo half-life, alter RNA affinity, increase nuclease resistance etc. [eg. see Agrawal & Iyer (1995) Cwrr Opin Biotechnol 6:12-19; Agrawal (1996) TIBTECH 14:376-387]; analogues such as peptide nucleic acids may also be used [eg. see Corey (1997) TIBTECH 15:224-229; Buchardt et al. (1993) TIBTECH 11:384-386]. Alternatively, the polymerase chain reaction (PCR) is another well-known means for detecting small amounts of target nucleic acids. The assay is described in: Mullis et al. [Meth. Enzymol. (1987) 155: 335-350]; US patents 4,683,195 and 4,683,202. Two "primer" nucleotides hybridize with the target nucleic acids and are used to prime the reaction. The primers can comprise sequence that does not hybridize to the sequence of the amplification target (or its complement) to aid with duplex stability or, for ex-ample, to incorporate a convenient restriction site. Typically, such sequence will flank the desired Neisserial sequence. A thermostable polymerase creates copies of target nucleic acids from the primers using the original target nucleic acids as a template. After a threshold amount of target nucleic acids are generated by the polymerase, they can be detected by more traditional methods, such as Southern blots. When using the Southern blot method, the labelled probe will hybridize to the Neisserial sequence (or its complement). Also, mRNA or cDNA can be detected by traditional blotting techniques described in Sambrook et al [supra]. mRNA, or cDNA generated from mRNA using a polymerase enzyme, can be purified and separated using gel electrophoresis. The nucleic acids on the gel are then blotted onto a solid support, such as nitrocellulose. The solid support is exposed to a labelled probe and then washed to remove any unhybridized probe. Next, the duplexes containing the labeled probe are detected. Typically, the probe is labelled with a radioactive moiety. BRIEF DESCRIPTION OF THE DRAWINGS Figures 1-20 show biochemical data obtained in the Examples, and also sequence analysis, for ORFs 37, 5, 2, 15, 22, 28, 32, 4, 61, 76, 89, 97, 106, 138, 23, 25, 27, 79, 85 and 132. Ml and M2 are molecular weight markers. Arrows indicate the position of the main recombinant product or, in Western blots, the position of the main N. meningitidis immunoreactive band. TP indicates N. meningitidis total protein extract; OMV indicates N. meningitidis outer membrane vesicle preparation. In bactericidal assay results: a diamond (♦) shows preimmune data; a triangle (A) shows GST control data; a circle (•) shows data with recombinant N. meningitidis protein. Computer .analyses show a hydrophilicity plot (upper), .an .antigenic index plot (middle), and an AMPHI analysis (lower). The AMPHI program has been used to predict T-cell epitopes [Gao et al. (1989) J. Immunol. 143:3007; Roberts et al. (1996) AIDS Res Hum Retrovir 12:593; Quakyi et al. (1992) Scand J Immunol suppl.l 1 :9) .and is available in the Protean package of DNASTAR, Inc. (1228 South Park Street, Madison, Wisconsin 53715 USA). EXAMPLES The examples describe nucleic acid sequences which have been identified in N. meningitidis, along with their putative translation products, and also those of N.gonorrhoeae. Not all of the nucleic acid sequences are complete ie. they encode less than the full-length wild-type protein. The examples are generally in the following format: • a nucleotide sequence which has been identified in N. meningitidis (strain B) • the putative translation product of this sequence • a computer analysis of the translation product based on database comparisons • corresponding gene and protein sequences identified in N. meningitidis (strain A) and in N.gonorrhoeae • a description of the characteristics of the proteins which indicates that they might be suitably antigenic • results of biochemical analysis (expression, purification, ELISA, FACS etc.) The ex>amples typically include details of sequence identity between species and strains. Proteins that are similar in sequence are generally similar in both structure and function, and the sequence identity often indicates a common evolutionary origin. Comparison with sequences of proteins of known function is widely used as a guide for the assignment of putative protein function to a new sequence and has proved particularly useful in whole-genome analyses. Sequence comparisons were performed at NCBI (http://www.ncbi.nlm.nih.gov) using the algorithms BLAST, BLAST2, BLASTn, BLASTp, tBLASTn, BLASTx, & tBLASTx [eg. see also Altschul et al. (1997) Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Research 25:2289-3402]. Searches were performed against the following databases: non-redundant GenBank+EMBL+DDBJ+PDB sequences and non-redundant GenBank CDS translations+PDB+SwissProt+SPupdate+PIR sequences. To comp.are Meningococcal and Gonococcal sequences, the tBLASTx algorithm was used, as implemented at http://www.genome.ou.edu gono_blast.html. The FASTA algorithm was also used to compare the ORFs (from GCG Wisconsin Package, version 9.0). Dots within nucleotide sequences (eg. position 495 in SEQ ID 11) represent nucleotides which have been arbitrarily introduced in order to maintain a reading frame. In the same way, double- underlined nucleotides were removed. Lower case letters (eg. position 496 in SEQ ID 11) represent ambiguities which arose during alignment of independent sequencing reactions (some of the nucleotide sequences in the examples are derived from combining the results of two or more experiments). Nucleotide sequences were scanned in all six reading frames to predict the presence of hydrophobic domains using an algorithm based on the statistical studies of Esposti et al. [Critical evaluation of the hydropathy of membrane proteins (1990) Ewr J Biochem 190:207-219]. These domains represent potential transmembrane regions or hydrophobic leader sequences. Open reading frames were predicted from fragmented nucleotide sequences using the program ORFFINDΕR (NCBI). Underlined amino acid sequences indicate possible transmembrane domains or leader sequences in the ORFs, as predicted by the PSORT algorithm (http://www.psort.nibb.ac.jp). Functional domains were also predicted using the MOTLFS program (GCG Wisconsin & PROSITΕ). Various tests can be used to assess the in vivo immunogencity of the proteins identified in the examples. For example, the proteins can be expressed recombinantly and used to screen patient sera by immunoblot. A positive reaction between the protein and patient serum indicates that the patient has previously mounted an immune response to the protein in question ie. the protein is -an immunogen. This method can also be used to identify immunodominant proteins. The recombinant protein can also be conveniently used to prepare antibodies eg. in a mouse. These can be used for direct confirmation that a protein is located on the cell-surface. Labelled antibody (eg. fluorescent labelling for FACS) can be incubated with intact bacteria and the presence of label on the bacterial surface confirms the location of the protein. In particular, the following methods (A) to (S) were used to express, purify and biochemically characterise the proteins of the invention: A) Chromosomal DNA preparation N. meningitidis strain 2996 was grown to exponential phase in 100ml of GC medium, harvested by centrifugation, and resuspended in 5ml buffer (20% Sucrose, 50mM Tris-HCl, 50mM EDTA, pH8). After 10 minutes incubation on ice, the bacteria were lysed by adding 10ml lysis solution (50mM NaCI, 1% Na-Sarkosyl, 50μg/ml Proteinase K), and the suspension was incubated at 37°C for 2 hours. Two phenol extractions (equilibrated to pH 8) and one ChCl 3 /isoamylalcohol (24:1) extraction were performed. DNA was precipitated by addition of 0.3M sodium acetate and 2 volumes ethanol, and was collected by centrifugation. The pellet was washed once with 70% ethanol and redissolved in 4ml buffer (lOmM Tris-HCl, ImM EDTA, pH 8). The DNA concentration was measured by reading the OD at 260 nm. B) Oligonucleotide design Synthetic oligonucleotide primers were designed on the basis of the coding sequence of each ORF, using (a) the meningococcus B sequence when available, or (b) the gonococcus/meningococcus A sequence, adapted to the codon preference usage of meningococcus as necessary. Any predicted signal peptides were omitted, by deducing the 5 '-end amplification primer sequence immediately downstream from the predicted leader sequence. For most ORFs, the 5' primers included two restriction enzyme recognition sites (BamHl-Ndel, BamHl-Nhel, or EcoRL-Nhel, depending on the gene's own restriction pattern); the 3' primers included a Xhol restriction site. This procedure was established in order to direct the cloning of each amplification product (corresponding to each ORF) into two different expression systems: pGEX-KG (using either BamHl-Xhol or EcoRl-Xhol), and pET21b+ (using either Ndel-Xhol or Nhel-Xhoϊ). 5 '-end primer tail: CGCGGATCCCATATG (BamHl-Ndel ) CGCGGATCCGCTAGC (BamHl-Nheϊ) CCGGAATTCTAGCTAGC (EcoRl-Nheϊ) 3'-end primer tail: CCCGCTCGAG (Xhol) For ORFs 5, 15, 17, 19, 20, 22, 27, 28, 65 & 89, two different amplifications were performed to clone each ORF in the two expression systems. Two different 5' primers were used for each ORF; the same 3 ' Xhol primer was used as before: 5'-end primer tail: GGAATTCCATATGGCCATGG (Ndel) 5 '-end primer tail: CGGGATCC (BamHl) ORF 76 was cloned in the pTRC expression vector and expressed as an amino-terminus His-tag fusion. In this particular case, the predicted signal peptide was included in the final product. Nhel- BamHl restriction sites were incorporated using primers: 5'-end primer tail: GATCAGCTAGCCATATG (Mel) 3 '-end primer tail: CGGGATCC (BamHl) As well as containing the restriction enzyme recognition sequences, the primers included nucleotides which hybridizeed to the sequence to be amplified. The number of hybridizing nucleotides depended on the melting temperature of the whole primer, and was determined for each primer using the formulae: T m = 4 (G+C)+ 2 (A+T) (tail excluded) T m = 64.9 + 0.41 (% GC) - 600/N (whole primer) The average melting temperature of the selected oligos were 65-70°C for the whole oligo and 50-55°C for the hybridising region alone. Table I (page 487) shows the forward and reverse primers used for each amplification. In certain cases, it will be noted that the sequence of the primer does not exactly match the sequence in the ORF. When initial amplifications were performed, the complete 5' and/or 3' sequence was not known for some meningococcal ORFs, although the corresponding sequences had been identified in gonococcus. For amplification, the gonococcal sequences could thus be used as the basis for primer design, altered to take account of codon preference. In particular, the following codons were changed: ATA→ATT; TCG→TCT; CAG→CAA; AAG→AAA; GAG→GAA; CGA→CGC; CGG→CGC; GGG→GGC. Italicised nucleotides in Table I indicate such a change. It will be appreciated that, once the complete sequence has been identified, this approach is generally no longer necessary. Oligos were synthesized by a Perkin Elmer 394 DNA/RNA Synthesizer, eluted from the columns in 2ml NH 4 OH, and deprotected by 5 hours incubation at 56°C. The oligos were precipitated by addition of 0.3M Na- Acetate and 2 volumes ethanol. The samples were then centrifuged and the pellets resuspended in either lOOμl or 1ml of water. OD 260 was determined using a Perkin Elmer Lambda Bio spectophotometer and the concentration was determined and adjusted to 2-10pmol/μl. C) Amplification The standard PCR protocol was as follows: 50-200ng of genomic DNA were used as a template in the presence of 20-40μM of each oligo, 400-800μM dNTPs solution, 1 x PCR buffer (including 1.5mM MgCl 2 ), 2.5 units TaqI DNA polymerase (using Perkin-Elmer AmpliTaQ, GIBCO Platinum, Pwo DNA polymerase, or Tahara Shuzo Taq polymerase). In some cases, PCR was optimsed by the addition of lOμl DMSO or 50μl 2M betaine. After a hot start (adding the polymerase during a preliminary 3 minute incubation of the whole mix at 95°C), each sample underwent a double-step amplification: the first 5 cycles were performed using as the hybridization temperature the one of the oligos excluding the restriction enzymes tail, followed by 30 cycles performed according to the hybridization temperature of the whole length oligos. The cycles were followed by a final 10 minute extension step at 72°C. The standard cycles were as follows: The elongation time varied according to the length of the ORF to be amplified. The amplifications were performed using either a 9600 or a 2400 Perkin Elmer GeneAmp PCR System. To check the results, 1/10 of the amplification volume was loaded onto a 1-1.5% agarose gel and the size of each amplified fragment compared with a DNA molecular weight marker. The amplified DNA was either loaded directly on a 1 % agarose gel or first precipitated with ethanol and resuspended in a suitable volume to be loaded on a 1% agarose gel. The DNA fragment corresponding to the right size band was then eluted and purified from gel, using the Qiagen Gel Extraction Kit, following the instructions of the manufacturer. The fmal volume of the DNA fragment was 30μl or 50μl of either water or lOmM Tris, pH 8.5. D) Digestion of PCR fragments The purified DNA corresponding to the amplified fragment was split into 2 aliquots and double- digested with: - N dell Xhol or NheUXhόl for cloning into pET-21b+ and further expression of the protein as a C-terminus His-tag fusion - BamHI/XhoI or EcoRI/XhoI for cloning into pGEX-KG and further expression of the protein as N-terminus GST fusion. - For ORF 76, Nhel/BamHl for cloning into pTRC-HisA vector and further expression of the protein as N-terminus His-tag fusion. - EcoRI/PstI, EcoRl/Sall, Sall/PstI for cloning into pGex-His .and further expression of the protein as N-terminus His-tag fusion Each purified DNA fragment was incubated (37°C for 3 hours to overnight) with 20 units of each restriction enzyme (New England Biolabs ) in a either 30 or 40μl final volume in the presence of the appropriate buffer. The digestion product was then purified using the QIAquick PCR purification kit, following the manufacturer's instructions, and eluted in a final volume of 30 or 50μl of either water or lOmM Tris-HCl, pH 8.5. The final DNA concentration was determined by 1% agarose gel electrophoresis in the presence of titrated molecular weight marker. E) Digestion of the cloning vectors (pET22B, pGEX-KG, pTRC-His A, and pGex-His) lOμg plasmid was double-digested with 50 units of each restriction enzyme in 200μl reaction volume in the presence of appropriate buffer by overnight incubation at 37°C. After loading the whole digestion on a 1% agarose gel, the band corresponding to the digested vector was purified from the gel using the Qiagen QIAquick Gel Extraction Kit and the DNA was eluted in 50μl of lOmM Tris-HCl, pH 8.5. The DNA concentration was evaluated by measuring OD 260 of the sample, and adjusted to 50μg/μl. lμl of plasmid was used for each cloning procedure. The vector pGEX-His is a modified pGEX-2T vector carrying a region encoding six histidine residues upstream to the thrombin cleavage site and containing the multiple cloning site of the vector pTRC99 (Pharmacia) . F) Cloning The fragments corresponding to each ORF, previously digested and purified, were ligated in both pET22b and pGEX-KG. In a final volume of 20μl, a molar ratio of 3:1 fragment/vector was ligated using 0.5μl of NEB T4 DNA ligase (400 units/μl), in the presence of the buffer supplied by the manufacturer. The reaction was incubated at room temperature for 3 hours. In some experiments, ligation was performed using the Boheringer "Rapid Ligation Kit", following the manufacturer's instructions. In order to introduce the recombinant plasmid in a suitable strain, lOOμl E. coli DH5 competent cells were incubated with the ligase reaction solution for 40 minutes on ice, then at 37°C for 3 minutes, then, after adding 800μl LB broth, again at 37°C for 20 minutes. The cells were then centrifuged at maximum speed in .an Eppendorf microfuge .and resuspended in approximately 200μl of the supernatant. The suspension was then plated on LB ampicillin (lOOmg/ml ). The screening of the recombinant clones was performed by growing 5 randomly-chosen colonies overnight at 37°C in either 2ml (pGEX or pTC clones) or 5ml (pET clones) LB broth + lOOμg/ml ampicillin. The cells were then pelletted and the DNA extracted using the Qiagen QIAprep Spin Miniprep Kit, following the manufacturer's instructions, to a final volume of 30μl. 5μl of each individual miniprep (approximately lg ) were digested with either Ndel/Xhol or BamHUXhol .and the whole digestion loaded onto a 1-1.5% agarose gel (depending on the expected insert size), in parallel with the molecular weight marker (1Kb DNA Ladder, GIBCO). The screening of the positive clones was made on the base of the correct insert size. For the cloning of ORFs 110, 111, 113, 115, 119, 122, 125 & 130, the double-digested PCR product was ligated into double-digested vector using EcoRl-Pstl cloning sites or, for ORFs 115 & 127, EcoRl-SaR or, for ORF 122, SaR-Pstl. After cloning, the recombin-ant plasmids were introduced in the E.coli host W3110. Individual clones were grown overnight at 37°C in L-broth with 50μl/ml ampicillin. G) Expression Each ORF cloned into the expression vector w.as transformed into the strain suitable for expression of the recombinant protein product, lμl of each construct was used to tr.ansform 30μl of E.coli BL21 (pGEX vector), E.coli TOP 10 (pTRC vector) ox E.coli BL21-DE3 (pET vector), as described above. In the case of the pGEX-His vector, the same E.coli strain (W3110) was used for initial cloning and expression. Single recombinant colonies were inoculated into 2ml LB+Amp (lOOμg/ml), incubated at 37°C overnight, then diluted 1:30 in 20ml of LB+Amp (lOOμg/ml) in 100ml flasks, making sure that the OD 600 ranged between 0.1 and 0.15. The flasks were incubated at 30°C into gyratory water bath shakers until OD indicated exponential growth suitable for induction of expression (0.4-0.8 OD for pET and pTRC vectors; 0.8-1 OD for pGEX and pGEX- His vectors). For the pET, pTRC and pGEX-His vectors, the protein expression was induced by addition of ImM LPTG, whereas in the case of pGEX system the final concentration of IPTG was 0.2mM. After 3 hours incubation at 30°C, the final concentration of the sample was checked by OD. In order to check expression, 1ml of each sample was removed, centrifuged in a microfuge, the pellet resuspended in PBS, and analysed by 12% SDS-PAGE with Coomassie Blue staining. The whole sample was centrifuged at 6000g and the pellet resuspended in PBS for further use. H) GST-fusion proteins large-scale purification. A single colony was grown overnight at 37°C on LB+Amp agar plate. The bacteria were inoculated into 20ml of LB+Amp liquid colture in a water bath shaker and grown overnight. Bacteria were diluted 1:30 into 600ml of fresh medium and allowed to grow at the optimal temperature (20-37°C) to OD 550 0.8-1. Protein expression was induced with 0.2mM LPTG followed by three hours incubation. The culture was centrifuged at 8000rpm at 4°C. The supernatant was discarded and the bacterial pellet was resuspended in 7.5ml cold PBS. The cells were disrupted by sonication on ice for 30 sec at 40W using a Branson sonifier B-15, frozen and thawed twice and centrifuged again. The supernatant was collected and mixed with 150μl Glutatione-Sepharose 4B resin (Ph.armacia) (previously washed with PBS) and incubated at room temperature for 30 minutes. The sample was centrifuged at 700 for 5 minutes at 4°C. The resin was washed twice with 10ml cold PBS for 10 minutes, resuspended in 1ml cold PBS, and loaded on a disposable column. The resin was washed twice with 2ml cold PBS until the flow-through reached OD 280 of 0.02-0.06. The GST-fusion protein was eluted by addition of 700μl cold Glutathione elution buffer (lOmM reduced glutathione, 50mM Tris-HCl) and fractions collected until the OD 280 was 0.1. 21μl of each fraction were loaded on a 12% SDS gel using either Biorad SDS-PAGE Molecular weight standard broad range (Ml) (200, 116.25, 97.4, 66.2, 45, 31, 21.5, 14.4, 6.5 kDa) or Amersham Rainbow Marker (M2) (220, 66, 46, 30, 21.5, 14.3 kDa) as standards. As the MW of GST is 26kDa, this value must be added to the MW of each GST-fusion protein. I) His-fusion solubility analysis (ORFs 111-129) To analyse the solubility of the His-fusion expression products, pellets of 3ml cultures were resuspended in buffer Ml [500μl PBS pH 7.2]. 25μl lysozyme (lOmg/ml) was added and the bacteria were incubated for 15 min at 4°C. The pellets were sonicated for 30 sec at 40W using a Branson sonifier B-15, frozen and thawed twice and then separated again into pellet and supernatant by a centrifugation step. The supernatant was collected and the pellet was resuspended in buffer M2 [8M urea, 0.5M NaCI, 20mM imidazole and 0.1M NaH 2 PO 4 ] and incubated for 3 to 4 hours at 4°C. After centrifugation, the supernatant was collected and the pellet was resuspended in buffer M3 [6M guanidinium-HCl, 0.5M NaCI, 20mM imidazole and 0.1M NaH 2 PO 4 ] overnight at 4°C. The supernatants from all steps were analysed by SDS-PAGE. The proteins expressed from ORFs 113, 119 and 120 were found to be soluble in PBS, whereas ORFs 111, 122, 126 and 129 need urea and ORFs 125 and 127 need guanidium-HCl for their solubilization. J) His-fusion large-scale purification. A single colony was grown overnight at 37°C on a LB + Amp agar plate. The bacteria were inoculated into 20ml of LB+Amp liquid culture and incubated overnight in a water bath shaker. Bacteria were diluted 1:30 into 600ml fresh medium and allowed to grow at the optimal temperature (20-37°C) to OD 550 0.6-0.8. Protein expression was induced by addition of ImM LPTG and the culture further incubated for three hours. The culture was centrifuged at 8000rpm at 4°C, the supernatant was discarded and the bacterial pellet was resuspended in 7.5ml of either (i) cold buffer A (300mM NaCI, 50mM phosphate buffer, lOmM imidazole, pH 8) for soluble proteins or (ii) buffer B (urea 8M, lOmM Tris-HCl, lOOmM phosphate buffer, pH 8.8) for insoluble proteins. The cells were disrupted by sonication on ice for 30 sec at 40W using a Branson sonifier B-15, frozen and thawed two times and centrifuged again. For insoluble proteins, the supernatant was stored at -20°C, while the pellets were resuspended in 2ml buffer C (6M guanidine hydrochloride, lOOmM phosphate buffer, lOmM Tris-HCl, pH 7.5) and treated in a homogenizer for 10 cycles. The product was centrifuged at 13000rpm for 40 minutes. Supernatants were collected and mixed with 150μl Ni + -resin (Pharmacia) (previously washed with either buffer A or buffer B, as appropriate) and incubated at room temperature with gentle agitation for 30 minutes. The sample was centrifuged at 700g for 5 minutes at 4°C. The resin was washed twice with 10ml buffer A or B for 10 minutes, resuspended in 1ml buffer A or B and loaded on a disposable column. The resin was washed at either (i) 4°C with 2ml cold buffer A or (ii) room temperature with 2ml buffer B, until the flow-through reached OD 280 of 0.02-0.06. The resin was washed with either (i) 2ml cold 20mM imidazole buffer (300mM NaCI, 50mM phosphate buffer, 20mM imidazole, pH 8) or (ii) buffer D (urea 8M, lOmM Tris-HCl, lOOmM phosphate buffer, pH 6.3) until the flow-through reached the O.D 280 of 0.02-0.06. The His-fusion protein was eluted by addition of 700μl of either (i) cold elution buffer A (300mM NaCI, 50mM phosphate buffer, 250mM imidazole, pH 8) or (ii) elution buffer B (urea 8M, lOmM Tris-HCl, lOOmM phosphate buffer, pH 4.5) and fractions collected until the O.D 280 was 0.1. 21μl of each fraction were loaded on a 12% SDS gel. K) His-fusion proteins renaturation 10% glycerol was added to the denatured proteins. The proteins were then diluted to 20μg/ml using dialysis buffer I (10% glycerol, 0.5M arginine, 50mM phosphate buffer, 5mM reduced glutathione, 0.5mM oxidised glutathione, 2M urea, pH 8.8) and dialysed against the same buffer at 4°C for 12- 14 hours. The protein was further dialysed against dialysis buffer II (10% glycerol, 0.5M arginine, 50mM phosphate buffer, 5mM reduced glutathione, 0.5mM oxidised glutathione, pH 8.8) for 12-14 hours at 4°C. Protein concentration was evaluated using the formula: Protein (mg/ml) = (1.55 x OD 280 ) - (0.76 x OD 260 ) L) His-fusion large-scale purification (ORFs 111-129) 500ml of bacterial cultures were induced and the fusion proteins were obtained soluble in buffer Ml, M2 or M3 using the procedure described above. The crude extract of the bacteria was loaded onto a Ni-NTA superflow column (Quiagen) equilibrated with buffer Ml, M2 or M3 depending on the solubilization buffer of the fusion proteins. Unbound material was eluted by washing the column with the same buffer. The specific protein was eluted with the corresponding buffer containing 500mM imidazole and dialysed against the corresponding buffer without imidazole. After each run the columns were sanitized by washing with at least two column volumes of 0.5 M sodium hydroxide and reequilibrated before the next use. M) Mice immunisations 20μg of each purified protein were used to immunise mice intraperitoneally. In the case of ORFs 2, 4, 15, 22, 27, 28, 37, 76, 89 and 97, Balb-C mice were immunised with Al(OH) 3 as adjuvant on days 1, 21 and 42, and immune response was monitored in samples taken on day 56. For ORFs 44, 106 and 132, CD1 mice were immunised using the same protocol. For ORFs 25 and 40, CD1 mice were immunised using Freund's adjuvant, rather than AL(OH) 3 , and the same immunisation protocol was used, except that the immune response was measured on day 42, rather than 56. Similarly, for ORFs 23, 32, 38 and 79, CD1 mice were immunised with Freund's adjuvant, but the immune response was measured on day 49. N) ELISA assay (sera analysis) The acapsulated MenB M7 strain was plated on chocolate agar plates and incubated overnight at 37°C. Bacterial colonies were collected from the agar plates using a sterile dracon swab and inoculated into 7ml of Mueller-Hinton Broth (Difco) containing 0.25% Glucose. Bacterial growth was monitored every 30 minutes by following OD 620 . The bacteria were let to grow until the OD reached the value of 0.3-0.4. The culture was centrifuged for 10 minutes at lOOOOrpm. The supernatant was discarded and bacteria were washed once with PBS, resuspended in PBS containing 0.025% formaldehyde, and incubated for 2 hours at room temperature and then overnight at 4°C with stirring. lOOμl bacterial cells were added to each well of a 96 well Greiner plate and incubated overnight at 4°C. The wells were then washed three times with PBT washing buffer (0.1% Tween-20 in PBS). 200μl of saturation buffer (2.7% Polyvinylpyrrolidone 10 in water) was added to each well and the plates incubated for 2 hours at 37°C. Wells were washed three times with PBT. 200μl of diluted sera (Dilution buffer: 1% BSA, 0.1% Tween-20, 0.1% NaN 3 in PBS) were added to each well and the plates incubated for 90 minutes at 37°C. Wells were washed three times with PBT. lOOμl of HRP-conjugated rabbit anti-mouse (Dako) serum diluted 1 :2000 in dilution buffer were added to each well and the plates were incubated for 90 minutes at 37°C. Wells were washed three times with PBT buffer. 1 OOμl of substrate buffer for HRP (25ml of citrate buffer pH5, lOmg of O-phenildiamine and lOμl of H 2 O) were added to each well and the plates were left at room temperature for 20 minutes. lOOμl H 2 SO 4 was added to each well and OD 490 was followed. The ELISA was considered positive when OD 490 was 2.5 times the respective pre-immune sera. O) FACScan bacteria Binding Assay procedure. The acapsulated MenB M7 strain was plated on chocolate agar plates and incubated overnight at 37°C. Bacterial colonies were collected from the agar plates using a sterile dracon swab and inoculated into 4 tubes containing 8ml each Mueller-Hinton Broth (Difco) containing 0.25% glucose. Bacterial growth was monitored every 30 minutes by following OD 620 . The bacteria were let to grow until the OD reached the value of 0.35-0.5. The culture was centrifuged for 10 minutes at 4000rpm. The supernatant was discarded and the pellet was resuspended in blocking buffer (1% BSA, 0.4% NaN 3 ) and centrifuged for 5 minutes at 4000rpm. Cells were resuspended in blocking buffer to reach OD 620 of 0.07. lOOμl bacterial cells were added to each well of a Costar 96 well plate. lOOμl of diluted (1:200) sera (in blocking buffer) were added to each well and plates incubated for 2 hours at 4°C. Cells were centrifuged for 5 minutes at 4000rpm, the supernatant aspirated and cells washed by addition of 200μl well of blocking buffer in each well. lOOμl of R- Phicoerytrin conjugated F(ab) 2 goat anti-mouse, diluted 1:100, was added to each well and plates incubated for 1 hour at 4°C. Cells were spun down by centrifugation at 4000rpm for 5 minutes and washed by addition of 200μl/well of blocking buffer. The supernatant was aspirated and cells resuspended in 200μl/well of PBS, 0.25% formaldehyde. Samples were transferred to FACScan tubes and read. The condition for FACScan setting were: FL1 on, FL2 and FL3 off; FSC-H threshold:92; FSC PMT Voltage: E 02; SSC PMT: 474; Amp. Gains 7.1; FL-2 PMT: 539; compensation values: 0. P) OMV preparations Bacteria were grown overnight on 5 GC plates, harvested with a loop and resuspended in 10 ml 20mM Tris-HCl. Heat inactivation was performed at 56°C for 30 minutes and the bacteria disrupted by sonication for 10 minutes on ice (50% duty cycle, 50% output). Unbroken cells were removed by centrifugation at 5000g for 10 minutes and the total cell envelope fraction recovered by centrifugation at 50000g at 4°C for 75 minutes. To extract cytoplasmic membrane proteins from the crude outer membranes, the whole fraction was resuspended in 2% sarkosyl (Sigma) and incubated at room temperature for 20 minutes. The suspension was centrifuged at lOOOOg for 10 minutes to remove aggregates, and the supernatant further ultracentrifuged at 50000g for 75 minutes to pellet the outer membranes. The outer membranes were resuspended in lOmM Tris-HCl, pH8 and the protein concentration measured by the Bio-Rad Protein assay, using BSA as a standard. Q) Whole Extracts preparation Bacteria were grown overnight on a GC plate, harvested with a loop and resuspended in 1ml of 20mM Tris-HCl. Heat inactivation was performed at 56°C for 30 minutes. R) Western blotting Purified proteins (500ng/lane), outer membrane vesicles (5μg) and total cell extracts (25μg) derived from MenB strain 2996 were loaded on 15% SDS-PAGE and transferred to a nitrocellulose membrane. The transfer was performed for 2 hours at 150mA at 4°C, in transferring buffer (0.3 % Tris base, 1.44 % glycine, 20% methanol). The membrane was saturated by overnight incubation at 4°C in saturation buffer (10% skimmed milk, 0.1% Triton XlOO in PBS). The membrane was washed twice with washing buffer (3% skimmed milk, 0.1% Triton XlOO in PBS) and incubated for 2 hours at 37°C with mice sera diluted 1 :200 in washing buffer. The membrane was washed twice and incubated for 90 minutes with a 1 :2000 dilution of horseradish peroxidase labelled anti- mouse Ig. The membrane was washed twice with 0.1%) Triton XlOO in PBS and developed with the Opti-4CN Substrate Kit (Bio-Rad). The reaction was stopped by adding water. S) Bactericidal assay MC58 strain was grown overnight at 37°C on chocolate agar plates. 5-7 colonies were collected and used to inoculate 7ml Mueller-Hinton broth. The suspension was incubated at 37°C on a nutator and let to grow until OD 620 was 0.5-0.8. The culture was aliquoted into sterile 1.5ml Eppendorf tubes and centrifuged for 20 minutes at maximum speed in a microfuge. The pellet was washed once in Gey's buffer (Gibco) and resuspended in the same buffer to an OD 620 of 0.5, diluted 1 :20000 in Gey's buffer and stored at 25°C. 50μl of Gey's buffer/1% BSA was added to each well of a 96-well tissue culture plate. 25μl of diluted mice sera (1 :100 in Gey's buffer/0.2% BSA) were added to each well and the plate incubated at 4°C. 25μl of the previously described bacterial suspension were added to each well. 25μl of either heat-inactivated (56°C waterbath for 30 minutes) or normal baby rabbit complement were added to each well. Immediately after the addition of the baby rabbit complement, 22μl of each sample/well were plated on Mueller-Hinton agar plates (time 0). The 96-well plate was incubated for 1 hour at 37°C with rotation and then 22μl of each sample/well were plated on Mueller-Hinton agar plates (time 1). After overnight incubation the colonies corresponding to time 0 and time 1 hour were counted. Table II (page 493) gives a summary of the cloning, expression and prurification results. Example 1 The following partial DNA sequence was identified in N. meningitidis <SEQ ID 1>: 1 ATGAAACAGA CAGTCAA.AT GCTTGCCGCC GCCCTGATTG CCTTGGGCTT 51 GAACCGACCG GTGTGGNCGG ATGACGTATC GGATTTTCGG GAAAACTTGC 101 A.GCGGCAGC ACAGGGAAAT GCAGCAGCCC AATACAATTT GGGCGCAATG 151 TAT.TACAAA GGACGCGCGT GCGCCGGGAT GATGCTGAAG CGGTCAGATG 201 GTATCGGCAG CCGGCGGAAC AGGGGTTAGC CCAAGCCCAA TACAATTTGG 251 GCTGGATGTA TGCCAACGGG CGCGC.GTGC GCCAAGATGA TACCGAAGCG 301 GTCAGATGGT ATCGGCAGGC GGCAGCGCAG GGGGTTGTCC AAGCCCAATA 351 CAATTTGGGC GTGATATATG CCGAAGGACG TGGAGTGCGC CAAGACGATG 401 TCGAAGCGGT CAGATGGTTT CGGCAGGCGG CAGCGCAGGG GGTAGCCCAA 451 GCCCAAAACA ATTTGGGCGT GATGTATGCC GAAAGANCGC GCGTGCGCCA 501 AGACCG... This corresponds to the amino acid sequence <SEQ LD 2; ORF37>: 1 MKQTVXMLAA ALIALGLNRP VWXDDVSDFR ENLXAAAQGN AAAQYNLGAM 51 YXQRTRVRRD DAEAVR YRQ PAEQGLAQAQ YNLG MYANG RXVRQDDTEA 101 VRWYRQAAAQ GWQAQYNLG VIYAEGRGVR QDDVEAVRWF RQAAAQGVAQ 151 AQNNLGVMYA ERXRVRQD... Further work revealed the complete nucleotide sequence <SEQ ID 3>: 1 ATGAAACAGA CAGTCAAATG GCTTGCCGCC GCCCTGATTG CCTTGGGCTT 51 GAACCGAGCG GTGTGGGCGG ATGACGTATC GGATTTTCGG GAAAACTTGC 101 AGGCGGCAGC ACAGGGAAAT GCAGCAGCCC AATACAATTT GGGCGCAATG 151 TATTACAAAG GACGCGGCGT GCGCCGGGAT GATGCTGAAG CGGTCAGATG 201 GTATCGGCAG GCGGCGGAAC AGGGGTTAGC CCAAGCCCAA TACAATTTGG 251 GCTGGATGTA TGCCAACGGG CGCGGCGTGC GCCAAGATGA TACCGAAGCG 301 GTCAGATGGT ATCGGCAGGC GGCAGCGCAG GGGGTTGTCC AAGCCCAATA 351 CAATTTGGGC GTGATATATG CCGAAGGACG TGGAGTGCGC CAAGACGATG 401 TCGAAGCGGT CAGATGGTTT CGGCAGGCGG CAGCGCAGGG GGTAGCCCAA 451 GCCCAAAACA ATTTGGGCGT GATGTATGCC GAAAGACGCG GCGTGCGCCA 501 AGACCGCGCC CTTGCACAAG AATGGTTTGG CAAGGCTTGT CAAAACGGAG 551 ACCAAGACGG CTGCGACAAT GACCAACGCC TGAAGGCGGG TTATTGA This corresponds to the amino acid sequence <SEQ ID 4; ORF37-l>: 1 MKQTVK LAA ALIALGLNRA VADDVSDFR ENLQAAAQGN AAAQYNLGAM 51 YYKGRGVRRD DAEAVR YRQ AAEQGLAQAQ YNLG MYANG RGVRQDDTEA 101 VRWYRQAAAQ GWQAQYNLG VIYAEGRGVR QDDVEAVRWF RQAAAQGVAQ 151 AQNNLGVMYA ERRGVRQDRA LAQEWFGKAC QNGDQDGCDN DQRLKAGY* Further work identified the corresponding gene in strain A of N. meningitidis <SEQ LD 5>: 1 ATGAAACAGA CAGTCAAATG GCTTGCCGCC GCCCTGATTG CCTTGGGCTT 51 GAACCAAGCG GTGTGGGCGG ATGACGTATC GGATTTTCGG GAAAACTTGC 101 AGGCGGCAGC ACAGGGAAAT GCAGCAGCCC AAAACAATTT GGGCGTGATG 151 TATGCCGAAA GACGCGGCGT GCGCCAAGAC CGCGCCCTTG CACAAGAATG 201 GCTTGGCAAG GCTTGTCAAA ACGGATACCA AGACAGCTGC GACAATGACC 251 AACGCCTGAA AGCGGGTTAT TGA This encodes a protein having amino acid sequence <SEQ ID 6; ORF37a>: 1 MKQTVKWLAA ALIALGLNQA VWADDVSDFR ENLQAAAQGN AAAQNNLGVM 51 YAERRGVRQD RALAQEWLGK ACQNGYQDSC DNDQRLKAGY * The originally-identified partial strain B sequence (ORF37) shows 68.0% identity over a 75aa overlap with ORF37a: 10 20 30 40 50 60 orf37.pep MKQTVXMLAAALIALGLNRPVWXDDVSDFRENLXAAAQGNAAAQYNLGAMYXQRTRVRRD I I I I I I I I I I I I I I I I: II I I I I I I I I I I I I I II I I I II 111:11 : I I I : I orf37a MKQTVKWLAAALIALGLNQAVWADDVSDFRENLQAAAQGNAAAQNNLGVMYAERRGVRQD 10 20 30 40 50 60 70 80 90 100 110 120 orf37. pep DAEAVRWYRQPAEQGLAQAQYNLGWMYANGRXVRQDDTEAVRWYRQAAAQGWQAQYNLG I I : I : : : I orf37a RALAQEWLGKACQNGYQDSCDNDQRLKAGYX 70 80 90 Further work identified the coπesponding gene in N.gonorrhoeae <SEQ ID 7 >: 1 ATGAAACAGA CAGTCAAATG GCTTGCCGCC GCCCTGATTG CCTTGGGCTT 51 GAACCAAGCG GTGTGGGCGG GTGACGTATC GGATTTTCGG GAAAACTTGC 101 AGgcggcaGA ACaggGAAAT GCAGCAGCCC AATTCAATTT GGGCGTGATG 151 TATGAAAATG GACAAGGAGT TCGTCAAGAT TATGTACAGG CAGTGCAGTG 201 GTATCGCAAG GCTTCAGAAC AAGGGGATGC CCAAGCCCAA TACAATTTGG 251 GCTTGATGTA TTACGATGGA CGCGGCGTGC GCCAAGACCT TGCGCTCGCT 301 CAACAATGGC TTGGCAAGGC TTGTCAAAAC GGAGACCAAA ACAGCTGCGA 351 CAATGACCAA CGCCTGAAGG CGGGTTATTA A This encodes a protein having amino acid sequence <SEQ ID 8; ORF37ng>: 1 MKQTVKWLAA ALIALGLNQA VWAGDVSDFR ENLQAAEQGN AAAQFNLGVM 51 YENGQGVRQD YVQAVQWYRK ASEQGDAQAQ YNLGLMYYDG RGVRQDLALA 101 QQWLGKACQN GDQNSCDNDQ RLKAGY* The originally-identified partial strain B sequence (ORF37) shows 64.9% identity over a 1 llaa overlap with ORF37ng: orf37.pep MKQTVXMLAAALIALGLNRPVWXDDVSDFRENLXAAAQGNAAAQYNLGAMYXQRTRVRRD 60 I I I I I I I I II I I I I I I: II I I I I II I II II 1111111:111:11 : I I: I orf37ng MKQTVKWLAAALIALGLNQAVWAGDVSDFRENLQAAEQGNAAAQFNLGVMYENGQGVRQD 60 orf37.pep DAEAVRWYRQPAEQGLAQAQYNLGWMYANGRXVRQDDTEAVRWYRQAAAQGWQAQYNLG 120 :: I I : I I I : : I I I I I I I I I I I I I : I I I I I I : I : I : I : I orf37ng YVQAVQWYRKASEQGDAQAQYNLGLMYYDGRGVRQDLALAQQWLGKACQNGDQNSCDNDQ 120 orf37.pep VIYAEGRGVRQDDVEAVRWFRQAAAQGVAQAQNNLGVMYAERXRVRQD 168 orf37ng RLKAGY 126 The complete strain B sequence (ORF37-1) and ORF37ng show 51.5% identity in 198 aa overlap: 10 20 30 40 50 60 orf37-1. pep MKQTVKWLAAALIALGLNRAVWADDVSDFRENLQAAAQGNAAAQYNLGAMYYKGRGVRRD I I I I I I I I I I I I I I I I I I: I I I I I I I I I I I I I I I I 1111111:111:11 : I: I I I : I orf37ng MKQTVKWLAAALIALGLNQAVWAGDVSDFRENLQAAEQGNAAAQFNLGVMYENGQGVRQD 10 20 30 40 50 60 70 80 90 100 110 120 orf37-1. pep DAEAVRWYRQAAEQGLAQAQYNLGWMYANGRGVRQDDTEAVRWYRQAAAQGWQAQYNLG :: I I: I I I: I: I I I I I I I I I I I I I : I I I I I I I orf37ng YVQAVQWYRKASEQGDAQAQYNLGLMYYDGRGVRQD 70 80 90 130 140 150 160 170 180 orf37-1.pep VIYAEGRGVRQDDVEAVRWFRQAAAQGVAQAQNNLGVMYAERRGVRQDRALAQEWFGKAC II I I: I : II II orf37ng LALAQQWLGKAC 100 190 199 orf37-1. pep QNGDQDGCDNDQRLKAGYX I I I I I :: I I I I I I I I I I I I orf37ng QNGDQNSCDNDQRLKAGYX 110 120 Computer analysis of these amino acid sequences indicates a putative leader sequence, and it was predicted that the proteins from N. meningitidis .and N.gonorrhoeae, and their epitopes, could be useful .antigens for vaccines or diagnostics, or for raising antibodies. ORF37-1 (l lkDa) was cloned in pET and pGex vectors and expressed in E.coli, as described above. The products of protein expression and purification were analyzed by SDS-PAGE. Figure IA shows the results of affinity purification of the GST-fusion protein, and Figure IB shows the results of expression of the His-fusion in E.coli. Purified GST-fusion protein was used to immunise mice, whose sera were used for ELISA (positive result), FACS analysis (Figure IC), .and a bactericidal assay (Figure ID). These experiments confirm that ORF37-1 is a surface-exposed protein, and that it is a useful immunogen. Figure IE shows plots of hydrophilicity, antigenic index, and AMPHI regions for ORF37-1. Example 2 The following partial DNA sequence was identified in N. meningitidis <SEQ LD 9>: TTCGGCGA CATCGGCGGT TTGAAGGTCA ATGCCCCCGT CAAATCCGCA GGCGTATTGG TCGGGCGCGT CGGCGCTATC GGACTTGACC CGAAATCCTA TCAGGCGAGG GTGCGCCTCG ATTTGGACGG CAAGTATCAG TTCAGCAGCG ACGTTTCCGC GCAAATCCTG ACTTCsGGAC TTTTGGGCGA GCAGTACATC GGGCTGCAGC AGGGCGGCGA CACGGAAAAC CTTGCTGCCG GCGACACCAT CTCCGTAACC AGTTCTGCAA TGGTTCTGGA AAACCTTATC GGCAAATTCA TGACGAGTTT TGCCGAGAAA AATGCCGACG GCGGCAATGC GGAAAAAGCC GCCGAATAA This corresponds to the amino acid sequence <SEQ LD 10>: 1 FGDIGGLKVN APVKSAGVLV GRVGAIGLDP KSYQARVRLD LDGKYQFSSD 51 VSAQILTSGL LGEQYIGLQQ GGDTENLAAG DTISVTSSAM VLENLIGKFM 101 TSFAEKNADG GNAEKAAE* Computer analysis of this amino acid sequence gave the following results: Homology with a hypothetical H.influenzae protein -fybrd.haein; accession number p45029 SEQ ID 9 and ybrd.haein show 48.4% aa identity in 122 aa overlap: 20 30 40 50 60 70 yrbd.h LGIGALVFLGLRVANVQGFAETKSYTVTATFDNIGGLKVRAPLKIGGWIGRVSAITLDE I:: I II I I I : II: I : I I : : I I I : I I : II N.m FGDIGGLKVNAPVKSAGVLVGRVGAIGLDP 10 20 30 80 90 100 110 120 130 yrbd.h KSYLPKVSIAINQEYNEIPENSSLSIKTSGLLGEQYIALTMGFDDGDTAMLKNGSQIQDT I I I :: |::::: : I ::::: I I I I I I I I I I I I : I I I I I : I : |: I I N.m KSYQARVRLDLDGKY-QFSSDVSAQILTSGLLGEQYIGLQQG GDTENLAAGDTISVT 40 50 60 70 80 140 150 160 yrbd.h TSAMVLEDLIGQFL—YGSKKSDGNEKSESTEQ : I I I I I I: I I I : I: : : : I : : I I : : :::: |: N.m SSAMVLENLIGKFMTSFAEKNADGGNAEKAAEX 90 100 110 120 Homology with a predicted ORF from N.gonorrhoeae SEQ LD 9 shows 99.2% identity over a ll 8aa overlap with a predicted ORF from N. gonorrhoeae: 20 30 40 50 60 70 yrbd GAAAVAFLAFRVAGGAAFGGSDKTYAVYADFGDIGGLKVNAPVKSAGVLVGRVGAIGLDP I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I N.m FGDIGGLKVNAPVKSAGVLVGRVGAIGLDP 10 20 30 80 90 100 110 120 130 yrbd KSYQARVRLDLDGKYQFSSDVSAQILTSGLLGEQYIGLQQGGDTENLAAGDTISVTSSAM I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I II I I I I I I I I I I I I I I I I I I I I I I I I I N.m KSYQARVRLDLDGKYQFSSDVSAQILTSGLLGEQYIGLQQGGDTENLAAGDTISVTSSAM 40 50 60 70 80 90 140 150 160 yrbd VLENLIGKFMTSFAEKNAEGGNAEKAAEX I I I I I I I I I I I I I I I I I I : I I I I II II I I N.m VLENLIGKFMTSFAEKNADGGNAEKAAEX 100 110 120 The complete yrbd H.influenzae sequence has a leader sequence and it is expected that the full- length homologous N. meningitidis protein will also have one. This suggests that it is either a membrane protein, a secreted protein, or a surface protein and that the protein, or one of its epitopes, could be a useful antigen for vaccines or diagnostics. Example 3 The following partial DNA sequence was identified in N. meningitidis <SEQ LD 11>: 1 ..ATTTTGATAT ACCTCATCCG CAAGAATCTA GGTTCGCCCG TCTTCTTCTT 51 TCAGGAACGC CCCGGAAAGG ACGGAAAACC TTTTAAAATG GTCAAATTCC 101 GTTCCATGCG CGACGGCTTG TATTCAGACG GCATTCCGCT GCCCGACGGA 151 GAACGCCTGA CACCGTTCGG CAAAAAACTG CGTGCCGcCA GTwTGGACGA 201 ACTGCCTGAA TTATGGAATA TCTTAAAAGG CGAGATGAGC CTGGTCGGCC 251 CCCGCCCGCT GCTGATGCAA TATCTGCCGC TGTACGACAA CTTCCAAAAC 301 CGCCGCCACG AAATGAAACC CGGCATTACC GGCTGGGCGC AGGTCAACGG 351 GCGCAACGCg CTTTCGTGGG ACGAAAAATT CGCCTGCGAT GTTTGGTATA 401 TCGACCACTT CAGCCTGTGC CTCGACATCA AAATCCTACT GCTGACGGTT 451 AAAAAAGTAT TAATCAAGGA AGGGATTTCC GCACAGGGCG AACA.aCCAT 501 GCCCCCTTTC ACAGGAAAAC GCAAACTCGC CGTCGTCGGT GCGGGCGGAC 551 ACGGAAAAGT CGTTGCCGAC CTTGCCGCCG CACTCGGCCG GTACAGGGAA 601 ATCGTTTTTC TGGACGACCG CGCACAAGGC AGCGTCAACG GCTTTTCCGT 651 CATCGGCACG ACGCTGCTGC TTGAAAACAG TTTATCGCCC GAACAATACG 701 ACGTCGCCGT CGCCGTCGGC AACAACCGCA TCCGCCGCCA AATCGCCGAA 751 AAAGCCGCCG CGCTCGGCTT CGCCCTGCCC GTACTGGTTC ATCCGGACGC 801 GACCGTCTCG CCTTCTGCAA CAGTCGGACA AGGCAGCGTC GTTATGGCGA 851 AAGCGGTCG.. This corresponds to the amino acid sequence <SEQ LD 12; ORF3>: 1 .. ILIYLIRKNL GSPVFFFQER PGKDGKPFKM VKFRSMRDGL YSDGIPLPDG 51 ERLTPFGKKL RAASXDELPE LWNILKGEMS LVGPRPLLMQ YLPLYDNFQN 101 RRHEMKPGIT GWAQVNGRNA LSWDEKFACD VWYIDHFSLC LDIKILLLTV 151 KKVLIKEGIS AQGEXTMPPF TGKRKLAWG AGGHGKWAD LAAALGRYRE 201 IVFLDDRAQG SVNGFSVIGT TLLLENSLSP EQYDVAVAVG NNRIRRQIAE 251 KAAALGFALP VLVHPDATVS PSATVGQGSV VMAKAV.. Further sequence analysis revealed the complete nucleotide sequence <SEQ ID 13>: 1 ATGAGTAAAT TCTTCAAACG CCTGTTTGAC ATTGTTGCCT CCGCCTCGGG 51 ACTGATTTTC CTCTCGCCAG TATTTTTGAT TTTGATATAC CTCATCCGCA 101 AGAATCTAGG TTCGCCCGTC TTCTTCTTTC AGGAACGCCC CGGAAAGGAC 151 GGAAAACCTT TTAAAATGGT CAAATTCCGT TCCATGCGCG ACGCGCTTGA 201 TTCAGACGGC ATTCCGCTGC CCGACGGAGA ACGCCTGACA CCGTTCGGCA 251 AAAAACTGCG TGCCGCCAGT TTGGACGAAC TGCCTGAATT ATGGAATATC 301 TTAAAAGGCG AGATGAGCCT GGTCGGCCCC CGCCCGCTGC TGATGCAATA 351 TCTGCCGCTG TACGACAACT TCCAAAACCG CCGCCACGAA ATGAAACCCG 401 GCATTACCGG CTGGGCGCAG GTCAACGGGC GCAACGCGCT TTCGTGGGAC 451 GAAAAATTCG CCTGCGATGT TTGGTATATC GACCACTTCA GCCTGTGCCT 501 CGACATCAAA ATCCTACTGC TGACGGTTAA AAAAGTATTA ATCAAGGAAG 551 GGATTTCCGC ACAGGGCGAA GCCACCATGC CCCCTTTCAC AGGAAAACGC 601 AAACTCGCCG TCGTCGGTGC GGGCGGACAC GGAAAAGTCG TTGCCGACCT 651 TGCCGCCGCA CTCGGCCGGT ACAGGGAAAT CGTTTTTCTG GACGACCGCG 701 CACAAGGCAG CGTCAACGGC TTTTCCGTCA TCGGCACGAC GCTGCTGCTT 751 GAAAACAGTT TATCGCCCGA ACAATACGAC GTCGCCGTCG CCGTCGGCAA 801 CAACCGCATC CGCCGCCAAA TCGCCGAAAA AGCCGCCGCG CTCGGCTTCG 851 CCCTGCCCGT TCTGGTTCAT CCGGACGCGA CCGTCTCGCC TTCTGCAACA 901 GTCGGACAAG GCAGCGTCGT TATGGCGAAA GCCGTCGTAC AGGCAGGCAG 951 CGTATTGAAA GACGGCGTGA TTGTGAACAC TGCCGCCACC GTCGATCACG 1001 ACTGCCTGCT TAACGCTTTC GTCCACATCA GCCCAGGCGC GCACCTGTCG 1051 GGCAACACGC ATATCGGCGA AGAAAGCTGG ATAGGCACGG GCGCGTGCAG 1101 CCGCCAGCAG ATCCGTATCG GCAGCCGCGC AACCATTGGA GCGGGCGCAG 1151 TCGTCGTACG CGACGTTTCA GACGGCATGA CCGTCGCGGG CAATCCGGCA 1201 AAGCCGCTGC CGCGCAAAAA CCCCGAGACC TCGACAGCAT AA This corresponds to the amino acid sequence <SEQ LD 14; ORF3-l>: 1 MSKFFKRLFD IVASASGLIF LSPVFLILIY LIRKNLGSPV FFFQERPGKD 51 GKPFKMVKFR SMRDALDSDG IPLPDGERLT PFGKKLRAAS LDELPELWNI 101 LKGEMSLVGP RPLLMQYLPL YDNFQNRRHE MKPGITGWAQ VNGRNALSWD 151 EKFACDVWYI DHFSLCLDIK ILLLTVKKVL IKEGISAQGE ATMPPFTGKR 201 KLAWGAGGH GKWADLAAA LGRYREIVFL DDRAQGSVNG FSVIGTTLLL 251 ENSLSPEQYD VAVAVGNNRI RRQIAEKAAA LGFALPVLVH PDATVSPSAT 301 VGQGSWMAK AWQAGSVLK DGVIVNTAAT VDHDCLLNAF VHISPGAHLS 351 GNTHIGEESW IGTGACSRQQ IRIGSRATIG AGAVWRDVS DGMTVAGNPA 401 KPLPRKNPET STA* Computer analysis of this amino acid sequence gave the following results: Homology with a predicted ORF from N.meninsitidis (strain A) ORF3 shows 93.0% identity over a 286aa overlap with an ORF (ORF3a) from strain A of N meningitidis: 10 20 30 orf3.pep ILIYLIRKNLGSPVFFFQERPGKDGKPFKMVKFR I I I I I I I I I I I I I I I I I I I II I I I I I I I I I I II I orf3a MSKFFKRLFDIVASASGLIFLSPVFLILIYLIRKNLGSPVFFFQERPGKDGKPFKMVKFR 10 20 30 40 50 60 40 50 60 70 80 90 or 3.pep SMRDGLYSDGIPLPDGERLTPFGKKLRAASXDELPELWNILKGEMSLVGPRPLLMQYLPL I I: I: I I I I I I I I I I I I I I I I I I I I I I I I I II I I I I : I I |: I I I I I I I I I I I I I II I orf 3a SMHDALDSDGILLPDGERLTPFGKKLRAASLDELPELWNVLKGDMSLVGPRPLLMQYLPL 70 80 90 100 110 120 100 110 120 130 140 150 orf 3 . pep YDNFQNRRHEMKPGITGWAQVNGRNALSWDEKFACDVWYIDHFSLCLDIKILLLTVKKVL I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I : I I I I : I I I I 1 I I I I I I I I I I I I I I I I I I orf 3a YDNFQNRRHEMKPGITGWAQVNGRNALSWDERFACDIWYIDHFSLCLDIKILLLTVKKVL 130 140 150 160 170 180 160 170 180 190 200 210 orf 3 . pep ^KEGISAQGEXTMPPFTGKRKLAWGAGGHGKWADLAAALGRYREIVFLDDRAQGSVNG I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I : I I I I I I I I I I I I I I I : I I I I I I orf 3a CEGISAQGEATMPPFTGKRKLAVVGAGGHGKVVAELAAALGTYGEIVFLDDRVQGSVNG 190 200 210 220 230 240 220 230 240 250 260 270 orf 3 . pep FSVIGTTLLLENSLSPEQYDVAVAVGNNRIRRQIAEKAAALGFALPVLVHPDATVSPSAT I I I I I I I I I I I I I I I I I : I : I I I I I I I I I I I I I I I I I I I I I I I I I I I : I I I : I I I I I I I orf3a FPVIGTTLLLENSLSPEQFDIAVAVGNNRIRRQIAEKAAALGFALPVLIHPDSTVSPSAT 250 260 270 280 290 300 280 orf3.pep VGQGSWMAKAV 1111:1111111 orf3a VGQGGWMAKAWQADSVLKDGVIVNTAATVDHDCLLDAFVHISPGAHLSGNTRIGEESW 310 320 330 340 350 360 The complete length ORF3a nucleotide sequence <SEQ LD 15> is: 1 ATGAGTAAAT TCTTCAAACG CCTGTTTGAC ATTGTTGCCT CCGCCTCGGG 51 ACTGATTTTC CTCTCGCCAG TATTTTTGAT TTTGATATAC CTCATCCGCA 101 AGAATCTGGG TTCGCCCGTC TTCTTCTTTC AGGAACGCCC CGGAAAGGAC 151 GGAAAACCTT TTAAAATGGT CAAATTCCGT TCCATGCACG ACGCGCTTGA 201 TTCAGACGGC ATTCTGCTGC CCGACGGAGA ACGCCTGACA CCGTTCGGCA 251 AAAAACTGCG TGCCGCCAGT TTGGACGAAC TGCCCGAACT GTGGAACGTC 301 CTCAAAGGCG ACATGAGCCT GGTCGGCCCC CGCCCGCTGC TGATGCAATA 351 TCTGCCGCTG TACGACAACT TCCAAAACCG CCGCCACGAA ATGAAACCGG 401 GCATTACCGG CTGGGCGCAG GTCAACGGGC GCAACGCGCT TTCGTGGGAC 451 GAACGCTTCG CATGCGACAT CTGGTATATC GACCACTTCA GCCTGTGCCT 501 CGACATCAAA ATCCTACTGC TGACGGTTAA AAAAGTATTA ATCAAAGAAG 551 GGATTTCCGC ACAGGGCGAA GCCACCATGC CCCCTTTCAC AGGAAAACGC 601 AAACTTGCCG TCGTCGGTGC GGGCGGACAC GGCAAAGTCG TTGCCGAGCT 651 TGCCGCCGCA CTCGGCACAT ACGGCGAAAT CGTTTTTCTG GACGACCGCG 701 TCCAAGGCAG CGTCAACGGC TTCCCCGTCA TCGGCACGAC GCTGCTGCTT 751 GAAAACAGTT TATCGCCCGA ACAATTCGAC ATCGCCGTCG CCGTCGGCAA 801 CAACCGCATC CGCCGCCAAA TCGCCGAAAA AGCCGCCGCG CTCGGCTTCG 851 CCCTGCCCGT CCTGATTCAT CCGGACTCGA CCGTCTCGCC TTCTGCAACA 901 GTCGGACAAG GCGGCGTCGT TATGGCGAAA GCCGTCGTAC AGGCTGACAG 951 CGTATTGAAA GACGGCGTAA TTGTGAACAC TGCCGCCACC GTCGATCACG 1001 ATTGCCTGCT TGATGCTTTC GTCCACATCA GCCCGGGCGC GCACCTGTCG 1051 GGCAACACGC GTATCGGCGA AGAAAGCTGG ATAGGCACAG GCGCGTGCAG 1101 CCGCCAGCAG ATCCGTATCG GCAGCCGCGC AACCATTGGA GCGGGCGCAG 1151 TCGTCGTGCG CGACGTTTCA GACGGCATGA CCGTCGCGGG CAACCCGGCA 1201 AAACCATTGG CAGGCAAAAA TACCGAGACC CTGCGGTCGT AA This is predicted to encode a protein having amino acid sequence <SEQ LD 16>: 1 MSKFFKRLFD IVASASGLIF LSPVFLILIY LIRKNLGSPV FFFQERPGKD 51 GKPFKMVKFR SMHDALDSDG ILLPDGERLT PFGKKLRAAS LDELPELWNV 101 LKGDMSLVGP RPLLMQYLPL YDNFQNRRHE MKPGITGWAQ VNGRNALSWD 151 ERFACDIWYI DHFSLCLDIK ILLLTVKKVL IKEGISAQGE ATMPPFTGKR 201 KLAWGAGGH GKWAELAAA LGTYGEIVFL DDRVQGSVNG FPVIGTTLLL 251 ENSLSPEQFD IAVAVGNNRI RRQIAEKAAA LGFALPVLIH PDSTVSPSAT 301 VGQGGWMAK AWQADSVLK DGVIVNTAAT VDHDCLLDAF VHISPGAHLS 351 GNTRIGEESW IGTGACSRQQ IRIGSRATIG AGAVWRDVS DGMTVAGNPA 401 KPLAGKNTET LRS* Two transmembrane domains are underlined. ORF3-1 shows 94.6% identity in 410 aa overlap with ORF3a: 10 20 30 40 50 60 orf3 . pep MSKFFKRLFDIVASASGLIFLSPVFLILIYLIRKNLGSPVFFFQERPGKDGKPFKMVKFR I I I I I I I I I I II I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I II I I I I I I I I I I I I I I I orf3-1 MSKFFKRLFDIVAΞASGLIFLSPVFLILIYLIRKNLGSPVFFFQERPGKDGKPFKMVKFR 10 20 30 40 50 60 70 80 90 100 110 120 orf3a .pep SMHDALDSDGILLPDGERLTPFGKKLRAASLDELPELWNVLKGDMSLVGPRPLLMQYLPL I I : I I I I I I I I I I I I I I I I I I I I I I I I II I I I I I I I I I : I I I : I I I I I I I I I I I I I I I I orf3-1 SMRDALDSDGIPLPDGERLTPFGKKLRAASLDELPELWNILKGEMSLVGPRPLLMQYLPL 70 80 90 100 110 120 130 140 150 160 170 180 orf3a . pep YDNFQNRRHEMKPGITGWAQVNGRNALSWDERFACDIWYIDHFSLCLDIKILLLTVKKVL I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I : I I I I : I I I I I I I I I I I I I I I I I I I I I I I orf 3-1 YDNFQNRRHEMKPGITGWAQVNGRNALSWDEKFACDVWYIDHFSLCLDIKILLLTVKKVL 130 140 150 160 170 180 190 200 210 220 230 240 orf 3a . pep IKEGISAQGEATMPPFTGKRKLAWGAGGHGKVVAELAAALGTYGEIVFLDDRVQGSVNG I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I : I I I I I I I I I I I I I I I : I I I I I I orf 3-1 IKEGISAQGEATMPPFTGKRKLAWGAGGHGKWADLAAALGRYREIVFLDDRAQGSVNG 190 200 210 220 230 240 250 260 270 280 290 300 orf 3a . pep FPVIGTTLLLENSLSPEQFDIAVAVGNNRIRRQIAEKAAALGFALPVLIHPDSTVSPSAT I I I I I I I I I I I I I I I I I : I : I I I I I I I I I I I I I I I I I I I I I I I I I I I : I I I : I I I I I I I orf3-1 FSVIGTTLLLENSLSPEQYDVAVAVGNNRIRRQIAEKAAALGFALPVLVHPDATVSPSAT 250 260 270 280 290 300 310 320 330 340 350 360 orf3a . pep VGQGGWMAKAWQADSVLKDGVIVNTAATVDHDCLLDAFVHISPGAHLSGNTRIGEESW I I I I : I I II I I I I I I I I I I I I I I I I I I I I I I I I I I I : I I I I I I I I I I I I I I I : I I I I I I orf3-1 VGQGSVVMAKAWQAGSVLKDGVIVNTAATVDHDCLLNAFVHISPGAHLSGNTHIGEESW 310 320 330 340 350 360 370 380 390 400 410 orf3a . pep IGTGACSRQQIRIGSRATIGAGAWVRDVSDGMTVAGNPAKPLAGKNTETLRSX I I I I I I I I I I I I I I I II I I I I I I I I I I I I I I I I I I I I I I I I I I II II orf3-l IGTGACSRQQIRIGSRATIGAGAVWRDVSDGMTVAGNPAKPLPRKNPETSTAX 370 380 390 400 410 Homology with hypothetical protein encoded by yvfc gene (accession Z719281 of B. subtilis ORF3 and YVFC proteins show 55% aa identity in 170 aa overlap (BLASTp): 0RF3 3 IYLIRKNLGSPVFFFQERPGKDGKPFKMVKFRSMRDGLYSDGIPLPDGERLTPFGKKLRA 62 I ++R +GSPVFF Q RPG GKPF + KFR+M D S G LPD RLT G+ +R yvfc 27 IAWRLKIGSPVFFKQVRPGLHGKPFTLYKFRTMTDERDSKGNLLPDEVRLTKTGRLIRK 86 0RF3 63 ASXDELPELWNILKGEMSLVGPRPLLMQYLPLYDNFQNRRHEMKPGITGWAQVNGRNALS 122 S DELP+L N+LKG++SLVGPRPLLM YLPLY Q RRHE+KPGITGWAQ+NGRNA+S yvfc 87 LSIDELPQLLNVLKGDLSLVGPRPLLMDYLPLYTEKQARRHEVKPGITGWAQINGRNAIS 146 ORF3 123 WDEKFACDVWYIDHFSLCLDXXXXXXXXXXXXXXEGISAQGEXTMPPFTG 172 W++KF DVWY+D++S LD EGI T FTG yvfc 147 WEKKFELDVWYVDNWSFFLDLKILCLTVRKVLVSEGIQQTNHVTAERFTG 196 Homology with a predicted ORF from N.sonorrhoeae ORF3 shows 86.3% identity over a 286aa overlap with a predicted ORF (ORF3.ng) from N gonorrhoeae: orf3 ILIYLIRKΝLGSPVFFFQERPGKDGKPFKMVKFR 34 : I I I I I I I I I I I I I I :: I I I I I I I I I I I I I I I I orf3ng MSKAVKRLFDIIASASGLIVLSPVFLVLIYLIRKΝKGSPVFFIRERPGKDGKPFKMVKFR 60 orf3 SMRDGLYSDGIPLPDGERLTPFGKKLRAASXDELPELWΝILKGEMSLVGPRPLLMQYLPL 94 I I I I: I I II I I I I I : I I I I I I I I I I I: I I I I I I I I I : I I I I I I I I I I II I I I I I I I I orf3ng SMRDALDSDGIPLPDSERLTDFGKKLRATSLDELPELWΝVLKGEMSLVGPRPLLMQYLPL 120 orf3 YDΝFQΝRRHEMKPGITGWAQVΝGRΝALSWDEKFACDVWYIDHFSLCLDIKILLLTVKKVL 154 I :: I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I : I I I I I I : I I : I I : I I I : I I I I I I I orf3ng YΝKFQΝRRHEMKPGITGWAQVΝGRΝALSWDEKFSCDVWYTDΝFSFWLDMKILFLTVKKVL 180 orf3 IKEGISAQGEXTMPPFTGKRKLAWGAGGHGKWADLAAALGRYREIVFLDDRAQGSVΝG 214 I I I I I I I I I I I I I I : I : I II I I : I I I I I I I I I I : I I I I I I I I I I I I I I I : I I I I I I orf3ng IKEGISAQGEATMPPFAGΝRKLAVIGAGGHGKWAELAAALGTYGEIVFLDDRTQGSVΝG 240 orf3 FSVIGTTLLLEΝSLSPEQYDVAVAVGΝΝRIRRQIAEKAAALGFALPVLVHPDATVSPSAT 274 I I I II I I I I I I I I I I I I : I:: I I II I I I I I I I I: I: I I I I II I I I I : I I I I I I I I II orf3ng FPVIGTTLLLEΝSLSPEQFDITVAVGΝΝRIRRQITEΝAAALGFKLPVLIHPDATVSPSAI 300 orf3 VGQGSVVMAKAV 286 : I I I I I I I I I I I orf3ng IGQGSWMAKAWQAGSVLKDGVIVΝTAATVDHDCLLDAFVHISPGAHLSGΝTRIGEESR 360 The complete length ORF3ng nucleotide sequence <SEQ ID 17> is: 1 ATGAGTAAAG CCGTCAAACG CCTGTTCGAC ATCATCGCAT CCGCATCGGG 51 GCTGATTGTC CTGTCGCCCG TGTTTTTGGT TTTAATATAC CTCATCCGCA 101 AAAACTTAGG TTCGCCCGTC TTCTTCattC GGGAACGCCc cgGAAAGGAc 151 ggaaaacCTT TTAAAATGGT CAAATTCCGT TCCAtgcgcg acgcgcttGA 201 TTCAGACGGC ATTCCGCTGC CCGATAGCGA ACGCCTGACC GATTTCGGCA 251 AAAAATTACG CGCCACCAGT TTGGACGAAC TTCCTGAATT ATGGAATGTC 301 CTCAAAGGCG AGATGAGCCT GGTCGGCCCC CGCCCGCTTT TGATGCAGTA 351 TCTGCCGCTT TACAACAAAT TTCAAAACCG CCGCCACGAA ATGAAACCGG 401 GCATTACCGG CTGGGCGCAG GTCAACGGGC GCAACGCGCT TTCGTGGGAC 451 GAAAAGTTCT CCTGCGATGT TTGGTACACC GACAATTTCA GCTTTTGGCT 501 GGATATGAAA ATCCTGTTTC TGACAGTCAA AAAAGTCTTG ATTAAAGAAG 551 GCATTTCGGC GCAAGGGGAA GCCACCATGC CCCCTTTCGC GGGGAATCGC 601 AAACTCGCCG TTATCGGCGC GGGCGGACAC GGCAAAGTCG TTGCCGAGCT 651 TGCCGCCGCA CTCGGCACAT ACGGCGAAAT CGTTTTTCTG GACGACCGCA 701 CCCAAGGCAG CGTCAACGGC TTCCCCGTCA TCGGCACGAC GCTGCTGCTT 751 GAAAACAGTT TATCGCCCGA ACAATTCGAC ATCACCGTCG CCGTCGGCAA 801 CAACCGCATC CGCCGCCAAA TCACCGAAAA CGCCGCCGCG CTCGGCTTCA 851 AACTGCCCGT TCTGATTCAT CCCGACGCGA CCGTCTCGCC TTCTGCAATA 901 ATCGGACAAG GCAGCGTCGT AATGGCGAAA GCCGTCGTAC AGGCCGGCAG 951 CGTATTGAAA GACGGCGTGA TTGTGAACAC TGCCGCCACC GTCGATCACG 1001 ACTGCCTGCT TGACGCTTTC GtccaCATCA GCCCGGGCGC GCACCTGTCG 1051 GGCAACACGC GTATCGGCGA AGAAAGCCGG ATAGGCACGG GCGCGTGCAG 1101 CCGCCAGCAG ACAACCGTCG GCAGCGGGGT TACCgccgGT GCAGGGgcGG 1151 TTATCGTATG CGACATCCCG GACGGCATGA CCGTCGCGGG CAACCCGGCA 1201 AAGCCCCTTA CGGGCAAAAA CCCCAAGACC GGGACGGCAT AA This encodes a protein having amino acid sequence <SEQ LD 18>: 1 MSKAVKRLFD IIASASGLIV LSPVFLVLIY LIRKΝLGSPV FFIRERPGKD 51 GKPFKMVKFR SMRDALDSDG IPLPDSERLT DFGKKLRATS LDELPELWΝV 101 LKGEMSLVGP RPLLMQYLPL YΝKFQΝRRHE MKPGITGWAQ VΝGRΝALSWD 151 EKFSCDVWYT DΝFSFWLDMK ILFLTVKKVL IKEGISAQGE ATMPPFAGΝR 201 KLAVIGAGGH GKWAELAAA LGTYGEIVFL DDRTQGSVΝG FPVIGTTLLL 251 EΝSLSPEQFD ITVAVGΝΝRI RRQITEΝAAA LGFKLPVLIH PDATVSPSAI 301 IGQGSWMAK AWQAGSVLK DGVIVΝTAAT VDHDCLLDAF VHISPGAHLS 351 GΝTRIGEESR IGTGACSRQQ TTVGSGVTAG AGAVIVCDIP DGMTVAGΝPA 401 KPLTGKΝPKT GTA* This protein shows 86.9% identity in 413 aa overlap with ORF3-1: 10 20 30 40 50 60 orf3-1.pep MSKFFKRLFDIVASASGLIFLSPVFLILIYLIRKNLGSPVFFFQERPGKDGKPFKMVKFR III I I I I I I : I I I I I I I I I I I I I : I I I I I I I I I I I I I I I : : I I I I I I I I I I I I I I I I orf3ng MSKAVKRLFDIIASASGLIVLSPVFLVLIYLIRKNLGSPVFFIRERPGKDGKPFKMVKFR 10 20 30 40 50 60 70 80 90 100 110 120 orf3-1.pep SMRDALDSDGIPLPDGERLTPFGKKLRAASLDELPELWNILKGEMSLVGPRPLLMQYLPL I I I I I I I I I I I I I I I : I I I I I II I I II : I I I I I I I I I I : I I I I I I I I I I I I I I I I II I I orf3ng SMRDALDSDGIPLPDSERLTDFGKKLRATSLDELPELWNVLKGEMSLVGPRPLLMQYLPL 70 80 90 100 110 120 130 140 150 160 170 180 orf3-1.pep YDNFQNRRHEMKPGITGWAQVNGRNALSWDEKFACDVWYIDHFSLCLDIKILLLTVKKVL I :: I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I : I I I I I I: I I: I I : I I I : I II I I I I orf3ng YNKFQNRRHEMKPGITGWAQVNGRNALSWDEKFSCDVWYTDNFSFWLDMKILFLTVKKVL 130 140 150 160 170 180 190 200 210 220 230 240 orf3-1.pep IKEGISAQGEATMPPFTGKRKLAWGAGGHGKWADLAAALGRYREIVFLDDRAQGSVNG I I I I II I I I I I I I I I I : I: I I I I I: I I I I I I I II I: I I I I I I I I I I I I I I I: II I I I I orf3ng IKEGISAQGEATMPPFAGNRKLAVIGAGGHGKWAELAAALGTYGEIVFLDDRTQGSVNG 190 200 210 220 230 240 250 260 270 280 290 300 orf 3-1 . pep FSVIGTTLLLENSLSPEQYDVAVAVGNNRIRRQIAEKAAALGFALPVLVHPDATVSPSAT I I I I I I I I I I I I I I I I I : I : : I I I I I I I I I I I I : I : I I I I I I I I I I : I I I I I I I I I I orf3ng FPVIGTTLLLENSLSPEQFDITVAVGNNRIRRQITENAAALGFKLPVLIHPDATVSPSAI 250 260 270 280 290 300 310 320 330 340 350 360 orf 3-1 . pep VGQGSWMAKAWQAGSVLKDGVIVNTAATVDHDCLLNAFVHISPGAHLSGNTHIGEESW : I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I : I I I I I I I I I I I I I I I : I I I I I orf3ng IGQGSWMAKAWQAGSVLKDGVIVNTAATVDHDCLLDAFVHISPGAHLSGNTRIGEESR 310 320 330 340 350 360 370 380 390 400 410 orf 3-1 . pep IGTGACSRQQIRIGSRATIGAGAWVRDVSDGMTVAGNPAKPLPRKNPETSTAX I I I I I I I I I I : I I : I I I I I I : I I : I I I I I I I I I I I I I I I I : I : I I I orf3ng IGTGACSRQQTTVGSGVTAGAGAVIVCDIPDGMTVAGNPAKPLTGKNPKTGTAX 370 380 390 400 410 In addition, ORF3ng shows significant homology with a hypothetical protein from B. subtilis: gnl|PID|e238668 (Z71928) hypothetical protein [Bacillus subtilis] >gi|1945702|gnl|PID|e313004 (Z94043) hypothetical protein [Bacillus subtilis] >gi|2635938|gnl|PID|ell86113 (Z99121) similar to capsular polysaccharide biosynthesis [Bacillus subtilis] Length = 202 Score = 235 bits (594), Expect = 3e-61 Identities = 114/195 (58%), Positives = 142/195 (72%) Query: 5 VKRLFDIIASASGLIVLSPVFLVLIYLIRKNLGSPVFFIRERPGKDGKPFKMVKFRSMRD 64 +KRLFD+ A+ L S + L I ++R +GSPVFF + RPG GKPF + KFR+M D Sbjct: 3 LKRLFDLTAAIFLLCCTSVIILFTIAVVRLKIGSPVFFKQVRPGLHGKPFTLYKFRTMTD 62 Query: 65 ALDSDGIPLPDSERLTDFGKKLRATSLDELPELWNVLKGEMSLVGPRPLLMQYLPLYNKF 124 DS G LPD RLT G+ +R S+DELP+L NVLKG++SLVGPRPLLM YLPLY + Sbjct: 63 ERDSKGNLLPDEVRLTKTGRLIRKLSIDELPQLLNVLKGDLSLVGPRPLLMDYLPLYTEK 122 Query: 125 QNRRHEMKPGITGWAQVNGRNALSWDEKFSCDVWYTDNFSFWLDMKILFLTVKKVLIKEG 184 Q RRHE+KPGITGWAQ+NGRNA+SW++KF DVWY DN+SF+LD+KIL LTV+KVL+ EG Sbjct: 123 QARRHEVKPGITGWAQINGRNAISWEKKFELDVWYVDNWSFFLDLKILCLTVRKVLVSEG 182 Query: 185 ISAQGEATMPPFAGN 199 I T F G+ Sbjct: 183 IQQTNHVTAERFTGS 197 The hypothetical product of yvfc gene shows similarity to EXOY of R.meliloti, an exopolysaccharide production protein. Based on this and on the two predicted transmembrane regions in the homologous N.gonorrhoeae sequence, it is predicted that these proteins, or their epitopes, could be useful antigens for vaccines or diagnostics, or for raising antibodies. Example 4 The following partial DNA sequence was identified in N. meningitidis <SEQ ID 19>: 1 ..AACCATATGG CGATTGTCAT CGACGAATAC GGCGGCACAT CCGGCTTGGT 51 CACCTTTGAA GACATCATCG AGCAAATCGT CGGCGAAATC GAAGACGAGT 101 TTGACGAAGA CGATAGCGCC GACAATATCC ATGCCGTTTC TTCAGACACG 151 TGGCGCATCC ATGCAGCTAC CGAAATCGAA GACATCAACA CCTTCTTCGG 201 CACGGAATAC AGCATCGAAG AAGCCGACAC CATT . GGCGG CCTGGTCATT 251 CAAGAGTTGG GACATCTGCC CGTGCGCGGC GAAAAAGTCC TTATCGGCGG 301 TTTGCAGTTC ACCGTCGCAC GCGCCGACAA CCGCCGCCTG CATACGCTGA 351 TGGCGACCCG CGTGAAGTAA GC ACCGC CGTTTCTGCA 401 CAGTTTAG This corresponds to .amino acid sequence <SEQ ID 20; ORF5>: 1 ..NHMAIVIDEY GGTSGLVTFE DIIEQIVGEI EDEFDEDDSA DNIHAVSSDT 51 WRIHAATEIE DINTFFGTEY SIEEADTIXR PGHSRVGTSA RARRKSPYRR 101 FAVHRRTRRQ PPPAYADGDP REVS....XR RFCTV* Further sequence analysis revealed the complete DNA sequence to be <SEQ LD 21>: 1 ATGGACGGCG CACAACCGAA AACGAATTTT TTTGAACGCC TGATTGCCCG 51 ACTCGCCCGC GAACCCGATT CCGCCGAAGA CGTATTAAAC CTGCTTCGGC 101 AGGCGCACGA GCAGGAAGTT TTTGATGCGG ATACGCTTTT AAGATTGGAA 151 AAAGTCCTCG ATTTTTCCGA TTTGGAAGTG CGCGACGCGA TGATTACGCG 201 CAGCCGTATG AACGTTTTAA AAGAAAACGA CAGCATCGAG CGCATCACCG 251 CCTACGTTAT CGATACCGCC CATTCGCGCT TCCCCGTCAT CGGCGAAGAC 301 AAAGACGAAG TTTTGGGCAT TTTGCACGCC AAAGACCTGC TCAAATATAT 351 GTTTAACCCC GAGCAGTTCC ACCTCAAATC CATTCTCCGC CCCGCCGTCT 401 TCGTCCCCGA AGGCAAATCG CTGACCGCCC TTTTAAAAGA GTTCCGCGAA 451 CAGCGCAACC ATATGGCGAT TGTCATCGAC GAATACGGCG GCACATCCGG 501 CTTGGTCACC TTTGAAGACA TCATCGAGCA AATCGTCGGC GAAATCGAAG 551 ACGAGTTTGA CGAAGACGAT AGCGCCGACA ATATCCATGC CGTTTCTTCC 601 GAACGCTGGC GCATCCATGC AGCTACCGAA ATCGAAGACA TCAACACCTT 651 CTTCGGCACG GAATACAGCA GCGAAGAAGC CGACACCATT CGGCCTGGTC 701 ATTCAAGAGT TGGGACATCT GCCCGTGCGC GGCGAAAAAG TCCTTATCGG 751 CGGTTTGCAG TTCACCGTCG CACGCGCCGA CAACCGCCGC CTGCATACGC 801 TGATGGCGAC CCGCGTGAAG TAAGCACCGC CGTTTCTGCA CAGTTTAGGA 851 TGACGGTACG GGCGTTTTCT GTTTCAATCC GCCCCATCCG CCAAACATAA This corresponds to amino acid sequence <SEQ ID 22; ORF5-l>: 1 MDGAQPKTNF FERLIARLAR EPDSAEDVLN LLRQAHEQEV FDADTLLRLE 51 KVLDFSDLEV RDAMITRSRM NVLKENDSIE RITAYVIDTA HSRFPVIGED 101 KDEVLGILHA KDLLKYMFNP EQFHLKSILR PAVFVPEGKS LTALLKEFRE 151 QRNHMAIVID EYGGTSGLVT FEDIIEQIVG EIEDEFDEDD SADNIHAVSS 201 ERWRIHAATE IEDINTFFGT EYSSEEADTI RPGHSRVGTS ARARRKSPYR 251 RFAVHRRTRR QPPPAYADGD PREVSTAVSA QFRMTVRAFS VSIRPIRQT* Further work identified the corresponding gene in strain A of N. meningitidis <SEQ LD 23 >: 1 ATGGACGGCG CACAACCGAA AACAAATTTT TTΝΝAACGCC TGATTGCCCG 51 ACTCGCCCGC GAACCCGATT CCGCCGAAGA CGTATTGACC CTGTTGCGCC 101 AAGCGCACGA ACAGGAAGTA TTTGATGCGG ATACGCTTTT AAGATTGGAA 151 AAAGTCCTCG ATTTTTCTGA TTTGGAAGTG CGCGACGCGA TGATTACGCG 201 CAGCCGTATG AACGTTTTAA AAGAAAACGA CAGCATCGAA CGCATCACCG 251 CCTACGTTAT CGATACCGCC CATTCGCGCT TCCCCGTCAT CGGTGAAGAC 301 AAAGACGAAG TTTTGGGTAT TTTGCACGCC AAAGACCTGC TCAAATATAT 351 GTTCAACCCC GAGCAGTTCC ACCTCAAATC GATATTGCGC CCTGCCGTCT 401 TCGTCCCCGA AGGCAAATCG CTGACCGCCC TTTTAAAAGA GTTCCGCGAA 451 CAGCGCAACC ATATGGCAAT CGTCATCGAC GAATACGGCG GCACGTCGGG 501 TTTGGTAACT TTTGAAGACA TCATCGAGCA AATCGTCGGC GACATCGAAG 551 ATGAGTTTGA CGAAGACGAA AGCGCGGACA ACATCCACGC CGTTTCCGCC 601 GAACGCTGGC GCATCCACGC GGCTACCGAA ATCGAAGACA TCAACGCCTT 651 TTTCGGCACG GAATACAGCA GCGAAGAAGC CGACACCATC GGCGGCCNTG 701 GTCATTCAGG AATTGGNACA CCTGCCCGTG CGCGGCGAAA AAGTCNTTAT 751 CGGCGNNTTG CANTTCACNG TCGCCNGCGC NGACAACCGC CGCCTGCATA 801 CGCTGATGGC GACCCGCGTG AAGTAAGCTC CGCCGTTTCT GTACAGTTTA 851 GGATGACGGT ACGGGCGTTT TCTGTTTCAA TCCGCCCCAT CCGCCANACA 901 TAA This encodes a protein having amino acid sequence <SEQ LD 24; ORF5a>: 1 MDGAQPKTNF XXR IARLAR EPDSAEDVLT LLRQAHEQEV FDADTLLRLE 51 KVLDFSDLEV RDAMITRSRM NVLKENDSIE RITAYVIDTA HSRFPVIGED 101 KDEVLGILHA KDLLKYMFNP EQFHLKSILR PAVFVPEGKS LTALLKEFRE 151 QRNHMAIVID EYGGTSGLVT FEDIIEQIVG DIEDEFDEDE SADNIHAVSA 201 ERWRIHAATE IEDINAFFGT EYSSEEADTI GGXGHSGIGT PARARRKSXY 251 RRXAXHXRXR XQPPPAYADG DPREVSSAVS VQFRMTVRAF SVSIRPIRXT 301 * The originally-identified partial strain B sequence (ORF5) shows 54.7% identity over a 124aa overlap with ORF5a: 10 20 30 orf5. pep NHMAIVIDEYGGTSGLVTFEDIIEQIVGEI I I I I I I I I I I I I I I II I I I I I I I I I I I I : I orf5a FHLKSILRPAVFVPEGKSLTALLKEFREQRNHMAIVIDEYGGTSGLVTFEDIIEQIVGDI 130 140 150 160 170 180 40 50 60 70 80 90 orf5. pep EDEFDEDDSADNIHAVSSDTWRIHAATEIEDINTFFGTEYSIEEADTIXRPGHSRVGTSA I I I II I I: I I I I I I I I I :: I I I I I I I I I I I I I : I I I I I I I I I I I I I III : I I I orf5a EDEFDEDESADNIHAVSAERWRIHAATEIEDINAFFGTEYSSEEADTIGGXGHSGIGTPA 190 200 210 220 230 240 100 110 120 130 orf5. pep RARRKSPYRRFAVHRRTRRQPPPAYADGDPREVSXXXXXRRFCTV II I I I I III I I i I I I I I I I I I I I I I I I orf5a RARRKSXYRRXAXHXRXRXQPPPAYADGDPREVSSAVSVQFRMTVRAFSVSIRPIRXTX 250 260 270 280 290 300 The complete strain B sequence (ORF5-1) and ORF5a show 92.7% identity in 300 aa overlap: 10 20 30 40 50 60 orf5a . pep MDGAQPKTNFXXRLIARLAREPDSAEDVLTLLRQAHEQEVFDADTLLRLEKVLDFSDLEV I I II I I II I I II I I I II I I I I I I I I I I: I I I II I I I I I I II II I I I I I I I I I I I I I I I orf5-1 MDGAQPKTNFFERLIARLAREPDSAEDVLNLLRQAHEQEVFDADTLLRLEKVLDFSDLEV 10 20 30 40 50 60 70 80 90 100 110 120 orf5a . pep RDAMITRSRMNVLKENDSIERITAYVIDTAHSRFPVIGEDKDEVLGILHAKDLLKYMFNP I II I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I orf5-1 RDAMITRSRMNVLKENDSIERITAYVIDTAHSRFPVIGEDKDEVLGILHAKDLLKYMFNP 70 80 90 100 110 120 130 140 150 160 170 180 orf5a.pep EQFHLKSILRPAVFVPEGKSLTALLKEFREQRNHMAIVIDEYGGTSGLVTFEDIIEQIVG I I I I I I I I I I I I I I I I I I I I I I I II I I I I II I I II I I I I I I I I I I I I I I I I I I I I I I I I I orf5-1 EQFHLKSILRPAVFVPEGKSLTALLKEFREQRNHMAIVIDEYGGTSGLVTFEDIIEQIVG 130 140 150 160 170 180 190 200 210 220 230 240 orf5a . pep DIEDEFDEDESADNIHAVSAERWRIHAATEIEDINAFFGTEYSSEEADTIGGXGHSGIGT : I I I I I I I I: I I I I I I I I I: I I I I I I I I I I I I I I I: I M I I I I I I I I I I I III : I I orf5-1 EIEDEFDEDDSADNIHAVSSERWRIHAATEIEDINTFFGTEYSSEEADTIRP-GHSRVGT 190 200 210 220 230 250 260 270 280 290 300 orf5a . pep PARARRKSXYRRXAXHXRXRXQPPPAYADGDPREVSSAVSVQFRMTVRAFSVSIRPIRXT I I I I I I I III I I I : I I I I I I I I I II I I I I I : I I I : I I I I I I I I I I I I I I I I I I orf5-1 SARARRKSPYRRFAVHRRTRRQPPPAYADGDPREVSTAVSAQFRMTVRAFSVSIRPIRQT 240 250 260 270 280 290 Further work identified the a partial DNA sequence in N.gonorrhoeae <SEQ LD 25> which encodes a protein having amino acid sequence <SEQ ID 26; ORF5ng>: 1 MDGAQPKTNF FERLIARLAR EPDSAEDVLN LLRQAHEQEV FDADTLTRLE 51 KVLDFAELEV RDAMITRSRM NVLKENDSIE RITAYVIDTA HSRFPVIGED 101 KDEVLGILHA KDLLKYMFNP EQFHLKSVLR PAVFVPEGKS LTALLKEFRE 151 QRNHMAIVID EYGGTSGLVT FEDIIEQIVG DIEDEFDEDE SADDIHSVSA 201 ERWRIHAATE IEDINAFFGT EYGSEEADTI RRLGHSGIGT PARARRKSPY 251 RRFAVHRRPR RQPPPAHADG DPREVSRACP HRRFCTV* Further analysis revealed the complete gonococcal nucleotide sequence <SEQ LD 27> to be: 1 ATGGACGGCG CACAACCGAA AACAAATTTT TTTGAACGCC TGATTGCCCG 51 ACTCGCCCGC GAACCCGATT CCGCCGAAGA CGTATTAAAC CTGCTTCGGC 101 AGGCGCACGA ACAGGAAGTT TTTGATGCCG ACACACTGAC CCGGCTGGAA 151 AAAGTATTGG ACTTTGCCGA GCTGGAAGTG CGCGATGCGA TGATTACGCG 201 CAGCCGCATG AACGTATTGA AAGAAAACGA CAGCATCGAA CGCATCACCG 251 CCTACGTCAT CGATACCGCC CATTCGCGCT TCCCCGTCAT CGGCGAAGAC 301 AAAGACGAAG TTTTGGGCAT TTTGCACGCC AAAGACCTGC TCAAATATAT 351 GTTCAACCCC GAGCAGTTCC ACCTGAAATC CGTCTTGCGC CCTGCCGTTT 401 TCGTGCCCGA AGGCAAATCT TTGACCGCCC TTTTAAAAGA GTTCCGCGAA 451 CAGCGCAACC ATATGGCAAT CGTCATCGAC GAATACGGCG GCACGTCGGG 501 TTTGGTCACC TTTGAAGACA TCATCGAGCA AATCGTCGGT GACATCGAAG 551 ACGAGTTTGA CGAAGACGAA AGCGccgacg acatCCACTC cgTTTccgCC 601 GAACGCTGGC GCATCCacgc ggctaCCGAA ATCGAAGaca TCAACGCCTT 651 TTTCGGTACG GAatacggca gcgaagaagc cgacaccatc cggcggctTG 701 GTCATTCAGG AATTGGGACA CCTGCCCGTG CGCGGCGAAA AAGTCCTTAt 751 cggcgGTTTG Cagttcaccg tCGCCCGCGC CGACAACCGC CGCCTGCACA 801 CGCTGATGGC GACCCGCGTG AAGTAAGCAG AGCCTGCCcg AccgccgttT 851 CTGCacAGTT TAGGatgACG gtaCGGTCGT TTTCTGTTTC AATCCGCCCC 901 ATCCGCCAAA CATAA This encodes a protein having amino acid sequence <SEQ ID 28; ORF5ng-l>: 1 MDGAQPKTNF FERLIARLAR EPDSAEDVLN LLRQAHEQEV FDADTLTRLE 51 KVLDFAELEV RDAMITRSRM NVLKENDSIE RITAYVIDTA HSRFPVIGED 101 KDEVLGILHA KDLLKYMFNP EQFHLKSVLR PAVFVPEGKS LTALLKEFRE 151 QRNHMAIVID EYGGTSGLVT FEDIIEQIVG DIEDEFDEDE SADDIHSVSA 201 ERWRIHAATE IEDINAFFGT EYGSEEADTI RRLGHSGIGT PARARRKSPY 251 RRFAVHRRPR RQPPPAHADG DPREVSRACP TAVSAQFRMT VRSFSVSIRP 301 IRQT* The originally-identified partial strain B sequence (ORF5) shows 83.1% identity over a 135aa overlap with the partial gonococcal sequence (ORF5ng): orf5 NHMAIVIDEYGGTSGLVTFEDIIEQIVGEI 30 I I I I I I I I I I I I I I I I I I I I I I I I I I I I : I orf5ng FHLKSVLRPAVFVPEGKSLTALLKEFREQRNHMAIVIDEYGGTSGLVTFEDIIEQIVGDI 182 orf5 EDEFDEDDSADNIHAVSSDTWRIHAATEIEDINTFFGTEYSIEEADTIXRPGHSRVGTSA 90 I I I I I I 1:1 I 1:11:1 I:: I I I I I I I I I I I I I : I I I I I I : I II I I I I III :| I I orfδng EDEFDEDESADDIHSVSAERWRIHAATEIEDINAFFGTEYGSEEADTIRRLGHSGIGTPA 242 orf5 RARRKSPYRRFAVHRRTRRQPPPAYADGDPREVSX RRFCTV 131 I I I I I I I I I I I I I I I I I I I I I II: I I I I I I I I I MINI orf5ng RARRKSPYRRFAVHRRPRRQPPPAHADGDPREVSRACPHRRFCTV 287 The complete strain B and gonococcal sequences (ORF5-1 8c ORF5ng-l) show 92.4% identity in 304 aa overlap: 10 20 30 40 50 60 orf5ng-l . pep MDGAQPKTNFFERLIARLAREPDSAEDVLNLLRQAHEQEVFDADTLTRLEKVLDFAELEV I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I:: I I I orf5-1 MDGAQPKTNFFERLIARLAREPDSAEDVLNLLRQAHEQEVFDADTLLRLEKVLDFSDLEV 10 20 30 40 50 60 70 80 90 100 110 120 orf5ng-l.pep RDAMITRSRMNVLKENDSIERITAYVIDTAHSRFPVIGEDKDEVLGILHAKDLLKYMFNP I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I II I I I I I I I I I I I I I I I I I I I I I I I orf5-1 RDAMITRSRMNVLKENDSIERITAYVIDTAHSRFPVIGEDKDEVLGILHAKDLLKYMFNP 70 80 90 100 110 120 130 140 150 160 170 180 orfδng-l.pep EQFHLKSVLRPAVFVPEGKSLTALLKEFREQRNHMAIVIDEYGGTSGLVTFEDIIEQIVG I I I I I I I: I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I II I orf5-1 EQFHLKSILRPAVFVPEGKSLTALLKEFREQRNHMAIVIDEYGGTSGLVTFEDIIEQIVG 130 140 150 160 170 180 190 200 210 220 230 240 orf5ng-l . pep DIEDEFDEDESADDIHSVSAERWRIHAATEIEDINAFFGTEYGSEEADTIRRLGHSGIGT : I I I I I I I I : I I I : I I : I I : I I I I I I I I I I I I I I I : I I I I I I : I I I I I I I I III : I I orf5-1 EIEDEFDEDDSADNIHAVSSERWRIHAATEIEDINTFFGTEYSSEEADTIRP-GHSRVGT 190 200 210 220 230 250 260 270 280 290 300 orf5ng-l . pep PARARRKSPYRRFAVHRRPRRQPPPAHADGDPREVSRACPTAVSAQFRMTVRSFSVSIRP I I I I I M I I I I I I I I I I I I I I I I I : I I I I I I I I I I I I I I I I I I I I I : I I I I I I I orf5-1 SARARRKSPYRRFAVHRRTRRQPPPAYADGDPREVS TAVSAQFRMTVRAFSVSIRP 240 250 260 270 280 290 orfδng-l.pep IRQTX I I I I I orf5-l IRQTX 300 Computer analysis of these amino acid sequences indicates a putative leader sequence, and identified the following homologies: Homology with hemolysin homolog TlvC (accession U32716) of H.influenzae ORF5 and TlyC proteins show 58%) aa identity in 77 aa overlap (BLASTp). ORF5 2 HMAIVIDEYGGTSGLVTFEDIIEQIVGEIEDEFDEDDSADNIHAVSSDTWRIHAATEIED 61 HMAIV+DE+G SGLVT EDI+EQIVG+IEDEFDE++ AD I +S T+ + A T+I+D TlyC 166 HMAIWDEFGAVSGLVTIEDILEQIVGDIEDEFDEEEIAD-IRQLSRHTYAVRALTDIDD 224 ORF5 62 INTFFGTEYSIEEADTI 78 N F T++ EE DTI TlyC 225 FNAQFNTDFDDEEVDTI 241 ORF5ng-l also shows significant homology with TlyC: SCORES Initl: 301 Initn: 419 Opt: 668 Smith-Waterman score: 668; 45.9% identity in 242 aa overlap 10 20 30 40 50 orf5ng-l.pep MDGAQPKTNFFERLIARLAR-EPDSAEDVLNLLRQAHEQEVFDADTLTRLEK I I |: I:: I: : I : I :::::: I :::::::: | : | :| tlyc_haein MNDEQQNSNQSENTKKPFFQSLFGRFFQGELKNREELVEVIRDSEQNDLIDQNTREMIEG 10 20 30 40 50 60 60 70 80 90 100 109 orf5ng-l.pep VLDFAELEVRDAMITRSRMNVLKENDSIERITAYVIDTAHSRFPVIGE—DKDEVLGILH I::: I I I: I I I II II:: :::::::: : I : : I I I I I I I I : : I : I :::| I I I tlyc_haein VMEIAELRVRDIMIPRSQIIFIEDQQDLNTCLNTIIESAHSRFPVIADADDRDNIVGILH 70 80 90 100 110 120 110 120 130 140 150 160 orf 5ng-l . pep AKDLLKYMF-NPEQFHLKSVLRPAVFVPEGKSLTALLKEFREQRNHMAIVIDEYGGTSGL I I I I I I : : : I I I : I : I I I : I : I I I : I : : I | : I I : I I I I I I : I I : I : : | | | tlyc haein AKDLLKFLREDAEVFDLSSLLRPWIVPESKRVDRMLKDFRSERFHMAIWDEFGAVSGL 130 140 150 160 170 180 170 180 190 200 210 220 orfδng-l.pep VTFEDIIEQIVGDIEDEFDEDESADDIHSVSAERWRIHAATEIEDINAFFGTEYGSEEAD I I : I I I : I I I I I I I I I I I I I : I I I I ::: I : : :: I I : I: I: I I I : I:: : I I: I tlyc aein VTIEDILEQIVGDIEDEFDEEEIAD-IRQLSRHTYAVRALTDIDDFNAQFNTDFDDEEVD 190 200 210 220 230 230 240 250 260 270 280 orf5ng-l . pep TIRRLGHSGIG-TPARARRKSPYRRFAVHRRPRRQPPPAHADGDPREVSRACPTAVSAQF II I : : I I I: tlyc_haein TIGGLIMQTFGYLPKRGEEIILKNLQFKVTSADSRRLIQLRVTVPDEHLAEMNNVDEKSE 240 250 260 270 280 290 Homology with a hypothetical secreted protein from E.coli: ORF5a shows homology to a hypothetical secreted protein from E.coli: sp|P77392|YBEX_ECOLI HYPOTHETICAL 33.3 KD PROTEIN IN CUTE-ASNB INTERGENIC REGION >gi 11778577 (U82598) similar to H. influenzae [ Escherichia coli ] >gi 11786879 (AE000170) f292; This 292 aa ORF is 23% identical (9 gaps) to 272 residues of an approx. 440 aa protein YTFL_HAEIN SW: P44717 [ Escherichia coli ] Length = 292 Score = 212 bits (533), Expect = 3e-54 Identities = 112/230 (48%), Positives = 149/230 (64%), Gaps = 3/230 (1%) Query: 2 DGAQPKTNFXXRLIARLAR-EPDSAEDVLTLLRQAHEQEVFDADTLLRLEKVLDFSDLEV 60 D K F L+++L EP + +++L L+R + + +4- D DT LE V+D +D V Sbjct: 10 DTISNKKGFFSLLLSQLFHGEPKNRDELLALIRDSGQNDLIDEDTRDMLEGVMDIADQRV 69 Query: 61 RDAMITRSRMNVLKENDSIERITAYVIDTAHSRFPVIGEDKDEVLGILHAKDLLKYM-FN 119 RD MI RS+M LK N +++ +I++AHSRFPVI EDKD + GIL AKDLL +M + Sbjct: 70 RDIMIPRSQMITLKRNQTLDECLDVIIESAHSRFPVISEDKDHIEGILMAKDLLPFMRSD 129 Query: 120 PEQFHLKSILRPAVFVPEGKSLTALLKEFREQRNHMAIVIDEYGGTSGLVTFEDIIEQIV 179 E F + +LR AV VPE K + +LKEFR QR HMAIVIDE+GG SGLVT EDI+E IV Sbjct: 130 AEAFSMDKVLRQAVVVPESKRVDRMLKEFRSQRYHMAIVIDEFGGVSGLVTIEDILELIV 189 Query: 180 GDIEDEFDEDESADNIHAVSAERWRIHAATEIEDINAFFGTEYSSEEADT 229 G+IEDE+DE++ D +S W + A IED N FGT +S EE DT Sbjct: 190 GEIEDEYDEEDDID-FRQLSRHTWTVRALASIEDFNEAFGTHFSDEEVDT 238 Based on this analysis, including the amino acid homology to the TlyC hemolysin-homologue from H. influenzae (hemolysins are secreted proteins), it was predicted that the proteins from N. meningitidis and N.gonorrhoeae are secreted and could thus be useful antigens for vaccines or diagnostics. ORF5-1 (30.7kDa) was cloned in the pGex vector and expressed in E.coli, as described above. The products of protein expression and purification were analyzed by SDS-PAGE. Figure 2A shows the results of affinity purification of the GST-fusion protein. Purified GST-fusion protein was used to immunise mice, whose sera were used for Western blot analysis (Figure IB). These experiments confirm that ORF5-1 is a surface-exposed protein, and that it is a useful immunogen. Example 5 The following partial DNA sequence was identified in N. meningitidis <SEQ ID 29>: 1 ATGCGCGGCG GCAGGCCGGA TTCCGTTACC GTGCAGATTA TCGAAGGTTC 51 GCGTTTTTCG CATATGAGGA AAGTCATCGA CGCAACGCCC GACATCGGAC 101 ACGACACCAA AGGCTGGAGC AATGAAAAAC TGATGGCGGA AGTTGCGCCC 151 GATGCCTTCA GCGGCAATCC TGAAgGGCAG TTTTTCCCCG ACAGCTACGA 201 AATCGATGCG GGCGGCAGTG ATTTGCAGAT TTACCAAACC GCCTACAAgG 251 GCGATGCAAC GCCGCCTGAA TGAgGGCATG GGAAAGCAGG CAGGACGGGC 301 TGCCTTATAA AAACCCTTAT GAAATGCTGA TTATGGCGAr CCTGGTCGAA 351 AAGGAAACAG GGCATGAAGC CGAsCsCGAC CATGTcGCTT CCGTCTTCGT 401 CAACCGCCTG AAAATCGGTA TGCGCCTGCA AACCgAssCG TCCGTGATTT 451 ACGGCATGGG TGCGGCATAC AAGGGCAAAA TCCGTAAAGC CGACCTGCGC 501 CGCGACACGC CGTACAACAC CTACACGCGC GGCGGTCTGC CGCCAACCCC 555511 GATTGCGCTG CCC.. This corresponds to the amino acid sequence <SEQ ID 30; ORF7>: 1 MRGGRPDSVT VQIIEGSRFS HMRKVIDATP DIGHDTKGWS NEKLMAEVAP 51 DAFSGNPEGQ FFPDSYEIDA GGSDLQIYQT AYKAMQRRLN EAWESRQDGL 101 PYKNPYEMLI MAXLVEKETG HEAXXDHVAS VFVNRLKIGM RLQTXXSVIY 151 GMGAAYKGKI RKADLRRDTP YNTYTRGGLP PTPIALP.. Further sequence analysis revealed the complete DNA sequence <SEQ ID 31>: 1 ATGTTGAGAA AATTGTTGAA ATGGTCTGCC GTTTTTTTGA CCGTGTCGGC 51 AGCCGTTTTC GCCGCGCTGC TTTTTGTTCC TAAGGATAAC GGCAGGGCAT 101 ACCGAATCAA AATTGCCAAA AACCAGGGTA TTTCGTCGGT CGGCAGGAAA 151 CTTGCCGAAG ACCGCATCGT GTTCAGCAGG CATGTTTTGA CGGCGGCGGC 201 CTACGTTTTG GGTGTGCACA ACAGGCTGCA TACGGGGACG TACAGATTGC 251 CTTCGGAAGT GTCTGCTTGG GATATCTTGC AGAAAATGCG CGGCGGCAGG 301 CCGGATTCCG TTACCGTGCA GATTATCGAA GGTTCGCGTT TTTCGCATAT 351 GAGGAAAGTC ATCGACGCAA CGCCCGACAT CGGACACGAC ACCAAAGGCT 401 GGAGCAATGA AAAACTGATG GCGGAAGTTG CGCCCGATGC CTTCAGCGGC 451 AATCCTGAAG GGCAGTTTTT CCCCGACAGC TACGAAATCG ATGCGGGCGG 501 CAGTGATTTG CAGATTTACC AAACCGCCTA CAAGGCGATG CAACGCCGCC 551 TGAATGAGGC ATGGGAAAGC AGGCAGGACG GGCTGCCTTA TAAAAACCCT 601 TATGAAATGC TGATTATGGC GAGCCTGGTC GAAAAGGAAA CAGGGCATGA 651 AGCCGACCGC GACCATGTCG CTTCCGTCTT CGTCAACCGC CTGAAAATCG 701 GTATGCGCCT GCAAACCGAC CCGTCCGTGA TTTACGGCAT GGGTGCGGCA 751 TACAAGGGCA AAATCCGTAA AGCCGACCTG CGCCGCGACA CGCCGTACAA 801 CACCTACACG CGCGGCGGTC TGCCGCCAAC CCCGATTGCG CTGCCCGGCA 851 AGGCGGCACT CGATGCCGCC GCCCATCCGT CCGGCGAAAA ATACCTGTAT 901 TTCGTGTCCA AAATGGACGG CACGGGCTTG AGCCAGTTCA GCCATGATTT 951 GACCGAACAC AATGCCGCCG TCCGCAAATA TATTTTGAAA AAATAA This corresponds to the amino acid sequence <SEQ ID 32; ORF7-l>: 1 MLRKLLKWSA VFLTVSAAVF AALLFVPKDN GRAYRIKIAK NQGISSVGRK 51 LAEDRIVFSR HVLTAAAYVL GVHNRLHTGT YRLPSEVSAW DILQKMRGGR 101 PDSVTVQIIE GSRFSHMRKV IDATPDIGHD TKGWSNEKLM AEVAPDAFSG 151 NPEGQFFPDS YEIDAGGSDL QIYQTAYKAM QRRLNEAWES RQDGLPYKNP 201 YEMLIMASLV EKETGHEADR DHVASVFVNR LKIGMRLQTD PSVIYGMGAA 251 YKGKIRKADL RRDTPYNTYT RGGLPPTPIA LPGKAALDAA AHPSGEKYLY 301 FVSKMDGTGL SQFSHDLTEH NAAVRKYILK K* Computer analysis of this amino acid sequence gave the following results: Homology with hypothetical protein encoded hy yceg gene (accession P44270) of H.influenzae ORF7 and yceg proteins show 44% aa identity in 192 aa overlap: ORF7 1 MRGGRPDSVTVQIIEGSRFSHMRKVIDATPDIGHDTKGWSNEKLMA EVAPDAFSG 55 + G+ V+ IEG F RK ++ P + K SNE++ A ++ + yceg 102 LNSGKEVQFNVKWIEGKTFKDWRKDLENAPHLVQTLKDKSNEEIFALLDLPDIGQNLELK 161 ORF7 56 NPEGQFFPDSYEIDAGGSDLQIYQTAYKAMQRRLNEAWESRQDGLPYKNPYEMLIMAXLV 115 N EG +PD+Y +DL++ + + + M++ LN+AW R + LP NPYEMLI+A +V yceg 162 NVEGWLYPDTYNYTPKSTDLELLKRSAERMKKALNKAWNERDEDLPLANPYEMLILASIV 221 ORF7 116 EKETGHEAXXDHVASVFVNRLKIGMRLQTXXSVIYGMGAAYKGKIRKADLRRDTPYNTYT 175 EKETG VASVF+NRLK M+LQT +VIYGMG Y G IRK DL TPYNTY yceg 222 EKETGIANERAKVASVFINRLKAKMKLQTDPTVIYGMGENYNGNIRKKDLETKTPYNTYV 281 0RF7 176 RGGLPPTPIALP 187 GLPPTPIA+P yceg 282 IDGLPPTPIAMP 293 The complete length YCEG protein has sequence: 1 MKKFLIAILL LILILAGVAS FSYYKMTEFV KTPVNVQADE LLTIERGTTS 51 SKLATLFEQE KLIADGKLLP YLLKLKPELN KIKAGTYSLE NVKTVQDLLD 101 LLNSGKEVQF NVKWIEGKTF KDWRKDLENA PHLVQTLKDK SNEEIFALLD 151 LPDIGQNLEL KNVEGWLYPD TYNYTPKSTD LELLKRSAER MKKALNKAWN 201 ERDEDLPLAN PYEMLILASI VEKETGIANE RAKVASVFIN RLKAKMKLQT 251 DPTVIYGMGE NYNGNIRKKD LETKTPYNTY VIDGLPPTPI AMPSESSLQA 301 VANPEKTDFY YFVADGSGGH KFTRNLNEHN KAVQEYLRWY RSQKNAK Homology with a predicted ORF from N. meningitidis (strain A) ORF7 shows 95.2% identity over a 187aa overlap with an ORF (ORF7a) from strain A of N meningitidis: 10 20 30 orf7.pep MRGGRPDSVTVQIIEGSRFSHMRKVIDATP I I I I I I I II I I I I I I I I I I I I I I I I I I I I I orf7a AAYVLGVHΝRLHTGTYRLPSEVSAWDILQKMRGGRPDSVTVQIIEGSRFSHMRKVIDATP 70 80 90 100 110 120 40 50 60 70 80 90 orf7. pep DIGHDTKGWSΝEKLMAEVAPDAFSGΝPEGQFFPDSYEIDAGGSDLQIYQTAYKAMQRRLΝ II I I I I I I I I I II I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I: I I I I I I I I I I I I I orf7a DIEHDTKGWSΝEKLMAEVAPDAFSGΝPEGQFFPDSYEIDAGGSDLRIYQIAYKAMQRRLΝ 130 140 150 160 170 180 100 110 120 130 140 150 orf7. pep EAWESRQDGLPYKΝPYEMLIMAXLVEKETGHEAXXDHVASVFVΝRLKIGMRLQTXXSVIY I I I I I I I I I I I I I I I I I I II I I I : I I I I I I I I I I I I I II I I I I II I I I I I I I I I I orf7a EAWESRQDGLPYKΝPYEMLIMASLIEKETGHEADRDHVASVFVΝRLKIGMRLQTDPSVIY 190 200 210 220 230 240 160 170 180 orf7. pep GMGAAYKGKIRKADLRRDTPYΝTYTRGGLPPTPIALP I I I I I I I I II I I I I I I I I I I I I I I I I I I I I I I I I I I I orf7a GMGAAYKGKIRKADLRRDTPYΝTYTRGGLPPTPIALPGKAALDAAAHPSGEKYLYFVSKM 250 260 270 280 290 300 orf7a DGTGLSQFSHDLTEHΝAAVRKYILKKX 310 320 330 The complete length ORF7a nucleotide sequence <SEQ ID 33> is: 1 ATGTTGAGAA AATTGTTGAA ATGGTCTGCC GTTTTTTTGA CCGTATCGGC 51 AGCCGTTTTC GCCGCGCTGC TTTTCGTCCC TAAAGACAAC GGCAGGGCAT 101 ACAGGATTAA AATTGCCAAA AACCAGGGTA TTTCGTCGGT CGGCAGGAAA 151 CTTGCCGAAG ACCGCATCGT GTTCAGCAGG CATGTTTTGA CGGCGGCGGC 201 CTACGTTTTG GGTGTGCACA ACAGGCTGCA TACGGGGACG TACAGACTGC 251 CTTCGGAAGT GTCTGCTTGG GATATCTTGC AGAAAATGCG CGGCGGCAGG 301 CCGGATTCCG TTACCGTGCA GATTATCGAA GGTTCGCGTT TTTCGCATAT 351 GAGGAAAGTC ATCGACGCAA CGCCCGACAT CGAACACGAC ACCAAAGGCT 401 GGAGCAATGA AAAACTGATG GCGGAAGTTG CCCCTGATGC CTTCAGCGGC 451 AATCCTGAAG GGCAGTTTTT CCCCGACAGC TACGAAATCG ATGCGGGCGG 501 CAGCGATTTA CGGATTTACC AAATCGCCTA CAAGGCGATG CAACGCCGAC 551 TGAATGAGGC ATGGGAAAGC AGGCAGGACG GGCTGCCTTA TAAAAACCCT 601 TATGAAATGC TGATTATGGC GAGCCTGATC GAAAAGGAAA CAGGGCATGA 651 AGCCGACCGC GACCATGTCG CTTCCGTCTT CGTCAACCGC CTGAAAATCG 701 GTATGCGCCT GCAAACCGAC CCGTCCGTGA TTTACGGCAT GGGTGCGGCA 751 TACAAGGGCA AAATCCGTAA AGCCGACCTG CGCCGCGACA CGCCGTACAA 801 CACCTACACG CGCGGCGGTC TGCCGCCAAC CCCGATCGCG CTGCCCGGCA 851 AGGCGGCACT CGATGCCGCC GCCCATCCGT CCGGTGAAAA ATACCTGTAT 901 TTCGTGTCCA AAATGGACGG TACGGGCTTG AGCCAGTTCA GCCATGATTT 951 GACCGAACAC AACGCCGCCG TTCGCAAATA TATTTTGAAA AAATAA This is predicted to encode a protein having amino acid sequence <SEQ LD 34>: 1 MLRKLLKWSA VFLTVSAAVF AALLFVPKDN GRAYRIKIAK NQGISSVGRK 51 LAEDRIVFSR HVLTAAAYVL GVHNRLHTGT YRLPSEVSAW DILQKMRGGR 101 PDSVTVQIIE GSRFSHMRKV IDATPDIEHD TKGWSNEKLM AEVAPDAFSG 151 NPEGQFFPDS YEIDAGGSDL RIYQIAYKAM QRRLNEAWES RQDGLPYKNP 201 YEMLIMASLI EKETGHEADR DHVASVFVNR LKIGMRLQTD PSVIYGMGAA 251 YKGKIRKADL RRDTPYNTYT RGGLPPTPIA LPGKAALDAA AHPSGEKYLY 301 FVSKMDGTGL SQFSHDLTEH NAAVRKYILK K* A leader peptide is underlined. ORF7a and ORF7-1 show 98.8% identity in 331 aa overlap: 10 20 30 40 50 60 orf7a . pep MLRKLLKWSAVFLTVSAAVFAALLFVPKDNGRAYRIKIAKNQGISSVGRKLAEDRIVFSR I I I I I I I I I I I I I I I I I I I I I I I II I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I orf7-1 MLRKLLKWSAVFLTVSAAVFAALLFVPKDNGRAYRIKIAKNQGISSVGRKLAEDRIVFSR 10 20 30 40 50 60 70 80 90 100 110 120 orf7a . pep HVLTAAAYVLGVHNRLHTGTYRLPSEVSAWDILQKMRGGRPDSVTVQIIEGSRFSHMRKV I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I II I I I I I I I I I I I I I I I I I I I orf7-1 HVLTAAAYVLGVHNRLHTGTYRLPSEVSAWDILQKMRGGRPDSVTVQIIEGSRFSHMRKV 70 80 90 100 110 120 130 140 150 160 170 180 orf7a . pep IDATPDIEHDTKGWSNEKLMAEVAPDAFSGNPEGQFFPDSYEIDAGGSDLRIYQIAYKAM I I I I I I I I I I I I I I I I I I I II I I I I I I I I I I I I I I I I I I I II I I I I I I I : II I I I I I I orf7-1 IDATPDIGHDTKGWSNEKLMAEVAPDAFSGNPEGQFFPDSYEIDAGGSDLQIYQTAYKAM 130 140 150 160 170 180 190 200 210 220 230 240 orf7a. pep QRRLNEAWESRQDGLPYKNPYEMLIMASLIEKETGHEADRDHVASVFVNRLKIGMRLQTD I II I I I II I I I I I I II I I I I I I I I I I I I I : I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I orf7-1 QRRLNEAWESRQDGLPYKNPYEMLIMASLVEKETGHEADRDHVASVFVNRLKIGMRLQTD 190 200 210 220 230 240 250 260 270 280 290 300 orf7a. pep PSVIYGMGAAYKGKIRKADLRRDTPYNTYTRGGLPPTPIALPGKAALDAAAHPSGEKYLY I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I II I II I I I I I I I I I I I I I I I I I I I I I I I I orf7-1 PSVIYGMGAAYKGKIRKADLRRDTPYNTYTRGGLPPTPIALPGKAALDAAAHPSGEKYLY 250 260 270 280 290 300 310 320 330 orf7a . pep FVSKMDGTGLSQFSHDLTEHNAAVRKYILKKX I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I orf7-1 FVSKMDGTGLSQFSHDLTEHNAAVRKYILKKX 310 320 330 Homology with a predicted ORF from N.gonorrhoeae ORF7 shows 94.7% identity over a 187aa overlap with a predicted ORF (ORF7.ng) from N. gonorrhoeae: orf7 MRGGRPDSVTVQIIEGSRFSHMRKVIDATPDIGHDTKGWSNEKLMAEVAPDAFSGNPEGQ 60 I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I II I I I I I I I I I II I I I I I I I I I I orf7ng MRGGRPDSVTVQIIEGSRFSHMRKVIDATPDIGHDTKGWSNEKLMAEVAPDAFSGNPEGQ 60 orf7 FFPDSYEIDAGGSDLQIYQTAYKAMQRRLNEAWESRQDGLPYKNPYEMLIMAXLVEKETG 120 I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I : I I I I I I I I I I I I I I I II I : I I I I I orf7ng FFPDSYEIDAGGSDLQIYQTAYKAMQRRLNEAWAGRQDGLPYKNPYEMLIMASLIEKETG 120 orf7 HEAXXDHVASVFVNRLKIGMRLQTXXSVIYGMGAAYKGKIRKADLRRDTPYNTYTRGGLP 180 III I I I I I I I I I I I I I I I I I I I I I I I I I I I I II I I I I I I I I I I I I I I I I I I I I I I orf7ng HEADRDHVASVFVNRLKIGMRLQTDPSVIYGMGAAYKGKIRKADLRRDTPYNTYTGGGLP 180 orf7 PTPIALP 187 II I I I I orf7ng PTRIALPGKAAMDAAAHPSGEKYLYFVSKMDGTGLΞQFSHDLTEHNAAVRKYILKK 236 An ORF7ng nucleotide sequence <SEQ LD 35> is predicted to encode a protein having amino acid sequence <SEQ ID 36>: 1 MRGGRPDSVT VQIIEGSRFS HMRKVIDATP DIGHDTKGWS NEKLMAEVAP 51 DAFSGNPEGQ FFPDSYEIDA GGSDLQIYQT AYKAMQRRLN EAWAGRQDGL 101 PYKNPYEMLI MASLIEKETG HEADRDHVAS VFVNRLKIGM RLQTDPSVIY 151 GMGAAYKGKI RKADLRRDTP YNTYTGGGLP PTRIALPGKA AMDAAAHPSG 201 EKYLYFVSKM DGTGLSQFSH DLTEHNAAVR KYILKK* Further sequence analysis revealed a partial DNA sequence of ORF7ng <SEQ ID 37>: 1 .. taccgaatca AGATTGCCAA AAATCAGGGT ATTTCGTCGG TCGGCAGGAA 51 ACTTGCcgaA GACCGCATCG TGTTCAGCAG GCATGTTTTG ACAGCGGCGG 101 CCTACGTTTT GGGTGTGCAC AACAGGCTGC ATACGGGGAC gTACAGATTG 151 CCTTCGGAAG TGTCTGCTTG GGATATCTTG CAGAAAATGC GCGGCGGCAG 201 GCCGGATTCC GTTACCGTGC AGATTATCGA AGGTTCGCGT TTTTCGCATA 251 TGAGGAAAGT CATCGACGCA ACGCCCGACA TCGGACACGA CACCAAAGGC 301 TGGAGCAATG AAAAACTGAT GGCGGAAGTT GCGCCCGATG CCTTCAGCGG 351 CAATCCTGAA GGGCAGTTTT TTCCCGACAG CTACGAAATC GATGCGGGCG 401 GCAGCGATTT GCAGATTTAC CAAACCGCCT ACAAGGCGAT GCAACGCCGC 451 CTGAACGAGG CATGGGCAGG CAGGCAGGAC GGGCTGCCTT ATAAAAACCC 501 TTATGAAATG CTGATTATGG CGAGCCTGAT CGAAAAGGAA ACGGGGCATG 551 AGGCCGACCG CGACCATGTC GCTTCCGTCT TCGTCAACCG CCTGAAAATC 601 GGTATGCGCC TGCAAACCGA CCCGTCCGTG ATTTACGGCA TGGGTGCGGC 651 ATACAAGGGC AAAATCCGTA AAGCCGACCT GCGCCGCGAC ACGCCGTACA 701 aCAccTAtac gggcgggggc ttgccgccaa cccggattgc gctgcccggC 751 Aaggcggcaa tggatgccgc cgcccacccg tccggcgaAa aatacctgTa 801 tttcgtgtcC AAAATGGACG GCACGGGCTT GAGCCAGTTC AGCCATGATT 851 TGACCGAACA CAACGCCGCc gTcCGCAAAT ATATTTTGAA AAAATAA pondis to the amino ;acid sequence <SEQ ID 38; ORF7ng- ■ 1>: 1 .. YRIKIAKNQG ISSVGRKLAE DRIVFSRHVL TAAAYVLGVH NRLHTGTYRL 51 PSEVSAWDIL QKMRGGRPDS VTVQIIEGSR FSHMRKVIDA TPDIGHDTKG 101 WSNEKLMAEV APDAFSGNPE GQFFPDSYEI DAGGSDLQIY QTAYKAMQRR 151 LNEAWAGRQD GLPYKNPYEM LIMASLIEKE TGHEADRDHV ASVFVNRLKI 201 GMRLQTDPSV IYGMGAAYKG KIRKADLRRD TPYNTYTGGG LPPTRIALPG 251 KAAMDAAAHP SGEKYLYFVS KMDGTGLSQF SHDLTEHNAA VRKYILKK* ORF7ng-l and ORF7-1 show 98.0% identity in 298 aa overlap: 10 20 30 40 50 60 orf7-1. pep KLLKWSAVFLTVSAAVFAALLFVPKDNGRAYRIKIAKNQGISSVGRKLAEDRIVFSRHVL I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I orf7ng-l YRIKIAKNQGISSVGRKLAEDRIVFSRHVL 10 20 30 70 80 90 100 110 120 orf7-1. pep TAAAYVLGVHNRLHTGTYRLPSEVSAWDILQKMRGGRPDSVTVQIIEGSRFSHMRKVIDA I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I II I I I I I I I I I I I I II I I I I orf7ng-l TAAAYVLGVHNRLHTGTYRLPSEVSAWDILQKMRGGRPDSVTVQIIEGSRFSHMRKVIDA 40 50 60 70 80 90 130 140 150 160 170 180 orf7-1. pep TPDIGHDTKGWSNEKLMAEVAPDAFSGNPEGQFFPDSYEIDAGGSDLQIYQTAYKAMQRR I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I orf ng-l TPDIGHDTKGWSNEKLMAEVAPDAFSGNPEGQFFPDSYEIDAGGSDLQIYQTAYKAMQRR 100 110 120 130 140 150 190 200 210 220 230 240 orf7-1. pep LNEAWESRQDGLPYKNPYEMLIMASLVEKETGHEADRDHVASVFVNRLKIGMRLQTDPSV IMM : I II II I M I I I I I II II I I: I I I I I I I I I I I I I I I I I I I I I I I I M I I II II I orf7ng-l LNEAWAGRQDGLPYKNPYEMLIMASLIEKETGHEADRDHVASVFVNRLKIGMRLQTDPSV 160 170 180 190 200 210 250 260 270 280 290 300 orf7-1.pep IYGMGAAYKGKIRKADLRRDTPYNTYTRGGLPPTPIALPGKAALDAAAHPSGEKYLYFVS I I I I II II I I I II I I I I I I I I II II II MMM II I II I II : II I I II I I II II II II orf7ng-l IYGMGAAYKGKIRKADLRRDTPYNTYTGGGLPPTRIALPGKAAMDAAAHPSGEKYLYFVS 220 230 240 250 260 270 310 320 330 orf7-1. pep KMDGTGLSQFSHDLTEHNAAVRKYILKKX I I II II II I II I I II II I I II I II II I II orf7ng-l KMDGTGLSQFSHDLTEHNAAVRKYILKKX 280 290 In addition, ORF7ng-l shows significant homology with a hypothetical E.coli protein: sp|P28306|YCEG_ECOLI HYPOTHETICAL 38.2 KD PROTEIN IN PABC-HOLB INTERGENIC REGION gi I 1787339 (AE000210) o340; 100% identical to fragment YCEG_ECOLI SW: P28306 but has 97 additional C-terminal residues [Escherichia coli] Length = 340 Score = 79 (36.2 bits), Expect = 5.0e-57, Sum P(2) = 5.0e-57 Identities = 20/87 (22%), Positives = 40/87 (45%) Query: 10 GISSVGRKLAEDRIVFSRHVLTAAAYVLGVHNRLHTGTYRLPSEVSAWDILQKMRGGRPD 69 G ++G +L D+I+ V + + GTYR +++ ++L+ + G+ Sbjct: 49 GRLALGEQLYADKIINRPRVFQWLLRIEPDLSHFKAGTYRFTPQMTVREMLKLLESGKEA 108 Query: 70 SVTVQIIEGSRFSHMRKVIDATPDIGH 96 ++++EG R S K + P I H Sbjct: 109 QFPLRLVEGMRLSDYLKQLREAPYIKH 135 Score = 438 (200.7 bits), Expect = 5.0e-57, Sum P(2) = 5.0e-57 Identities = 84/155 (54%), Positives = 111/155 (71%) Query: 120 EGQFFPDSYEIDAGGSDLQIYQTAYKAMQRRLNEAWAGRQDGLPYKNPYEMLIMASLIEK 179 EG F+PD++ A +D+ + + A+K M + ++ AW GR DGLPYK+ +++ MAS+IEK Sbjct: 158 EGWFWPDTWMYTANTTDVALLKRAHKKMVKAVDSAWEGRADGLPYKDKNQLVTMASIIEK 217 Query: 180 ETGHEADRDHVASVFVNRLKIGMRLQTDPSVIYGMGAAYKGKIRKADLRRDTPYNTYTGG 239 ET ++RD VASVF+NRL+IGMRLQTDP+VIYGMG Y GK+ +ADL T YNTYT Sbjct: 218 ETAVASERDKVASVFINRLRIGMRLQTDPTVIYGMGERYNGKLSRADLETPTAYNTYTIT 277 Query: 240 GLPPTRIALPGKAAMDAAAHPSGEKYLYFVSKMDG 274 GLPP IA PG ++ AAAHP+ YLYFV+ G Sbjct: 278 GLPPGAIATPGADSLKAAAHPAKTPYLYFVADGKG 312 Based on this analysis, including the fact that the H.influenzae YCEG protein possesses a possible leader sequence, it is predicted that the proteins from N meningitidis and N.gonorrhoeae, and their epitopes, could be useful antigens for vaccines or diagnostics, or for raising antibodies. Example 6 The following partial DΝA sequence was identified in N. meningitidis <SEQ ID 39>: 1 CGTTTCAAAA TGTTAACTGT GTTGACGGCA ACCTTGATTG CCGGACAGGT 51 ATCTGCCGCC GGAGGCGGTG CGGGGGATAT GAAACAGCCG AAGGAAGTCG 101 GAAAGGTTTT CAGAAAGCAG CAGCGTTACA GCGAGGAAGA AATCAAAAAC 151 GAACGCGCAC GGCTTGCGGC AGTGGGCGAG CGGGTTAATC AGATATTTAC 201 GTTGCTGGGA GGGGAAACCG CCTTGCAAAA GGGGCAGGCG GGAACGGCTC 251 TGGCAACCTA TATGCTGATG TTGGAACGCA CAAAATCCCC CGAAGTCGCC 301 GAACGCGCCT TGGAAATGGC CGTGTCGCTG AACGCGTTTG AACAGGCGGA 351 AATGATTTAT CAGAAATGGC GGCAGATTGA GCCTATACCG GGTAAGGCGC 401 AAAAACGGGC GGGGTGGCTG CGGAACGTGC TGAGGGAAAG AGGAAATCAG 451 CATCTGGACG GACGGGAAGA AGTGCTGGCT CAGGCGGACG AAGGACAG This corresponds to the amino acid sequence <SEQ ID 40; ORF9>: 1 ..RFKMLTVLTA TLIAGQVSAA GGGAGDMKQP KEVGKVFRKQ QRYSEEEIKΝ 51 ERARLAAVGE RVΝQIFTLLG GETALQKGQA GTALATYMLM LERTKSPEVA 101 ERALEMAVSL ΝAFEQAEMIY QKWRQIEPIP GKAQKRAGWL RΝVLRERGΝQ 151 HLDGREEVLA QADEGQ Further sequence .analysis revealed the complete DNA sequence <SEQ ID 41>: 1 ATGTTACCTA ACCGTTTCAA AATGTTAACT GTGTTGACGG CAACCTTGAT 51 TGCCGGACAG GTATCTGCCG CCGGAGGCGG TGCGGGGGAT ATGAAACAGC 101 CGAAGGAAGT CGGAAAGGTT TTCAGAAAGC AGCAGCGTTA CAGCGAGGAA 151 GAAATCAAAA ACGAACGCGC ACGGCTTGCG GCAGTGGGCG AGCGGGTTAA 201 TCAGATATTT ACGTTGCTGG GAGGGGAAAC CGCCTTGCAA AAGGGGCAGG 251 CGGGAACGGC TCTGGCAACC TATATGCTGA TGTTGGAACG CACAAAATCC 301 CCCGAAGTCG CCGAACGCGC CTTGGAAATG GCCGTGTCGC TGAACGCGTT 351 TGAACAGGCG GAAATGATTT ATCAGAAATG GCGGCAGATT GAGCCTATAC 401 CGGGTAAGGC GCAAAAACGG GCGGGGTGGC TGCGGAACGT GCTGAGGGAA 451 AGAGGAAATC AGCATCTGGA CGGACTGGAA GAAGTGCTGG CTCAGGCGGA 501 CGAAGGACAG AACCGCAGGG TGTTTTTATT GTTGGCACAA GCCGCCGTGC 551 AACAGGACGG GTTGGCGCAA AAAGCATCGA AAGCGGTTCG CCGCGCGGCG 601 TTGAAATATG AACATCTGCC CGAAGCGGCG GTTGCCGATG TGGTGTTCAG 651 CGTACAGGGA CGCGAAAAGG AAAAGGCAAT CGGAGCTTTG CAGCGTTTGG 701 CGAAGCTCGA TACGGAAATA TTGCCCCCCA CTTTAATGAC GTTGCGTCTG 751 ACTGCACGCA AATATCCCGA AATACTCGAC GGCTTTTTCG AGCAGACAGA 801 CACCCAAAAC CTTTCGGCCG TCTGGCAGGA AATGGAAATT ATGAATCTGG 851 TTTCCCTGCA CAGGCTGGAT GATGCCTATG CGCGTTTGAA CGTGCTGTTG 901 GAACGCAATC CGAATGCAGA CCTGTATATT CAGGCAGCGA TATTGGCGGC 951 AAACCGAAAA GAAGGTGCTT CCGTTATCGA CGGCTACGCC GAAAAGGCAT 1001 ACGGCAGGGG GACGGAGGAA CAGCGGAGCA GGGCGGCGCT AACGGCGGCG 1051 ATGATGTATG CCGACCGCAG GGATTACGCC AAAGTCAGGC AGTGGCTGAA 1101 AAAAGTATCC GCGCCGGAAT ACCTGTTCGA CAAAGGTGTG CTGGCGGCTG 1151 CGGCGGCTGT CGAGTTGGAC GGCGGCAGGG CGGCTTTGCG GCAGATCGGC 1201 AGGGTGCGGA AACTTCCCGA ACAGCAGGGG CGGTATTTTA CGGCAGACAA 1251 TTTGTCCAAA ATACAGATGC TCGCCCTGTC GAAGCTGCCC GATAAACGGG 1301 AGGCTTTGAG GGGGTTGGAC AAGATTATCG AAAAACCGCC TGCCGGCAGT 1351 AATACAGAGT TACAGGCAGA GGCATTGGTA CAGCGGTCAG TTGTTTACGA 1401 TCGGCTTGGC AAGCGGAAAA AAATGATTTC AGATCTTGAA AGGGCGTTCA 1451 GGCTTGCACC CGATAACGCT CAGATTATGA ATAATCTGGG CTACAGCCTG 1501 CTGACCGATT CCAAACGTTT GGACGAAGGT TTCGCCCTGC TTCAGACGGC 1551 ATACCAAATC AACCCGGACG ATACCGCTGT CAACGACAGC ATAGGCTGGG 1601 CGTATTACCT GAAAGGCGAC GCGGAAAGCG CGCTGCCGTA TCTGCGGTAT 1651 TCGTTTGAAA ACGACCCCGA GCCCGAAGTT GCCGCCCATT TGGGCGAAGT 1701 GTTGTGGGCA TTGGGCGAAC GCGATCAGGC GGTTGACGTA TGGACGCAGG 1751 CGGCACACCT TACGGGAGAC AAGAAAATAT GGCGGGAAAC GCTCAAACGT 1801 CACGGCATCG CATTGCCCCA ACCTTCCCGA AAACCTCGGA AATAA This corresponds to the amino acid sequence <SEQ ID 42; ORF9-l>: 1 MLPNRFKMLT VLTATLIAGQ VSAAGGGAGD MKQPKEVGKV FRKQQRYSEE 51 EIKNERARLA AVGERVNQIF TLLGGETALQ KGQAGTALAT YMLMLERTKS 101 PEVAERALEM AVSLNAFEQA EMIYQKWRQI EPIPGKAQKR AGWLRNVLRE 151 RGNQHLDGLE EVLAQADEGQ NRRVFLLLAQ AAVQQDGLAQ KASKAVRRAA 201 LKYEHLPEAA VADWFSVQG REKEKAIGAL QRLAKLDTEI LPPTLMTLRL 251 TARKYPEILD GFFEQTDTQN LSAVWQEMEI MNLVSLHRLD DAYARLNVLL 301 ERNPNADLYI QAAILAANRK EGASVIDGYA EKAYGRGTEE QRSRAALTAA 351 MMYADRRDYA KVRQWLKKVS APEYLFDKGV LAAAAAVELD GGRAALRQIG 401 RVRKLPEQQG RYFTADNLSK IQMLALSKLP DKREALRGLD KIIEKPPAGS 451 NTELQAEALV QRSWYDRLG KRKKMISDLE RAFRLAPDNA QIMNNLGYSL 501 LTDSKRLDEG FALLQTAYQI NPDDTAVNDS IGWAYYLKGD AESALPYLRY 551 SFENDPEPEV AAHLGEVLWA LGERDQAVDV WTQAAHLTGD KKIWRETLKR 601 HGIALPQPSR KPRK* Computer analysis of this amino acid sequence gave the following results: Homology with a predicted ORF from N. meningitidis (strain Al ORF9 shows 89.8% identity over a 166aa overlap with an ORF (ORF9a) from strain A of N meningitidis: 10 20 30 40 50 orf 9 . pep RFKMLTVLTATLIAGQVSAAGGGAGDMKQPKEVGKVFRKQQRYSEEEIKΝERARLA I I : I : I I : I : I I I I : I I I I : I I I I I I I I I I I I I I I I I I I I I I I I I I I I I orf 9a MLPARFTILSVLAAALLAGQAYAA— GAADAKPPKEVGKVFRKQQRYSEEEIKΝERARLA 10 20 30 40 50 60 70 80 90 100 110 orf9. pep AVGERVNQIFTLLGGETALQKGQAGTALATYMLMLERTKSPEVAERALEMAVSLNAFEQA II II I II II I I I II I I II II II I I I I I I I II II I II II I II II II II I I I II I I II II I orf9a AVGERVNQIFTLLGXETALQKGQAGTALATYMLMLERTKSPEVAERALEMAVSLNAFEQA 60 70 80 90 100 110 120 130 140 150 160 orf9. pep EMIYQKWRQIEPIPGKAQKRAGWLRNVLRERGNQHLDGREEVLAQADEGQ I II I II I I I I II I II II II II II I I I I I I I II II I II I II II II II I orf9a EMIYQKWRQIEPIPGKAQKRAGWLRNVLRERGNQHLDGLEEXLAQADEXQNRRVFLLLAQ 120 130 140 150 160 170 orf9a AAVQQDGLAQKASKAVRRAALRYEHLPEAAVADWFSVQXREKEKAIGALQRLAKLDTEI 180 190 200 210 220 230 The complete length ORF9a nucleotide sequence <SEQ ID 43> is: 1 ATGTTACCCG CCCGTTTCAC CATTTTATCT GTGCTCGCGG CAGCCCTGCT 51 TGCCGGGCAG GCGTATGCCG CCGGCGCGGC GGATGCGAAG CCGCCGAAGG 101 AAGTCGGAAA GGTTTTCAGA AAGCAGCAGC GTTACAGCGA GGAAGAAATC 151 AAAAACGAAC GCGCACGGCT TGCGGCAGTG GGCGAGCGGG TTAATCAGAT 201 ATTTACGTTG CTGGGANGGG AAACCGCCTT GCAAAAGGGG CAGGCGGGAA 251 CGGCTCTGGC AACCTATATG CTGATGTTGG AACGCACAAA ATCCCCCGAA 301 GTCGCCGAAC GCGCCTTGGA AATGGCCGTG TCNCTGAACG CGTTTGAACA 351 GGCGGAAATG ATTTATCAGA AATGGCGGCA GATTGAGCCT ATACCGGGTA 401 AGGCGCAAAA ACGGGCGGGG TGGCTGCGGA ACGTGCTGAG GGAAAGAGGA 451 AATCAGCATC TAGACGGACT GGAAGAANTG CTGGCTCAGG CGGACGAANG 501 ACAGAACCGC AGGGTGTTTT TATTGTTGGC ACAAGCCGCC GTGCAACAGG 551 ACGGGTTGGC GCAAAAAGCA TCGAAAGCGG TTCGCCGCGC GGCGTTGAGA 601 TATGAACATC TGCCCGAAGC GGCGGTTGCC GATGTGGTGT TCAGCGTACA 651 GGNACGCGAA AAGGAAAAGG CAATCGGAGC TTTGCAGCGT TTGGCGAAGC 701 TCGATACGGA AATATTGCCC CCCACTTTAA TGACGTTGCG TCTGACTGCA 751 CGCAAATATC CCGAAATACT CGACGGCTTT TTCGAGCAGA CAGACACCCA 801 AAACCTTTCG GCCGTCTGGC AGGAAATGGA AATTATGAAT CTGGTTTCCC 851 TGCACAGGCT GGATGATGCC TATGCGCGTT TGAACGTGCT GTTGGAACGC 901 AATCCGAATG CAGACCTGTA TATTCAGGCA GCGATATTGG CGGCAAACCG 951 AAAAGAANGT GCTTCCGTTA TCGACGGCTA CGCCGAAAAG GCATACGGCA 1001 GGGGGACGGG GGAACAGCGG GGCAGGGCGG CAATGACGGC GGCGATGATA 1051 TATGCCGACC GAAGGGATTA CACCAAAGTC AGGCAGTGGT TGAAAAAAGT 1101 GTCCGCGCCG GAATACCTGT TCGACAAAGG TGTGCTGGCG GCTGCGGCGG 1151 CTGTCGAGTT GGACNGCGGC AGGGCGGCTT TGCGGCAGAT CGGCAGGGTG 1201 CGGAAACTTC CCGAACAGCA GGGGCGGTAT TTTACGGCAG ACAATTTGTC 1251 CAAAATACAG ATGTTCGCCC TGTCGAAGCT GCCCGACAAA CGGGAGGCTT 1301 TGAGGGGGTT GGACAAGATT ATCGAAAAAC CGCCTGCCGG CAGTAATACA 1351 GAGTTACAGG CAGAGGCATT GGTACAGCGG TCAGTTGTTT ACGATCGGCT 1401 TGGCAAGCGG AAAAAAATGA TTTCAGATCT TGAAAGGGCG TTCAGGCTTG 1451 CACCCGATAA CGCTCAGATT ATGAATAATC TGGGCTACAG CCTGCTTTCC 1501 GATTCCAAAC GTTTGGACGA AGGCTTCGCC CTGCTTCAGA CGGCATACCA 1551 AATCAACCCG GACGATACCG CTGTCAACGA CAGCATAGGC TGGGCGTATT 1601 ACCTGAAANG CGACGCGGAA AGCGCGCTGC CGTATCTGCG GTATTCGTTT 1651 GAAAACGACC CCGAGCCCGA AGTTGCCGCC CATTTGGGCG AAGTGTTGTG 1701 GGCATTGGGC GAACGCGATC AGGCGGTTGA CGTATGGACG CAGGCGGCAC 1751 ACCTTACGGG AGACAAGAAA ATATGGCGGG AAACGCTCAA ACGTCACGGC 1801 ATCGCATTGC CCCAACCTTC CCGAAAACCT CGGAAATAA This encodes a protein having amino acid sequence <SEQ LD 44>: 1 MLPARFTILS VLAAALLAGQ AYAAGAADAK PPKEVGKVFR KQQRYSEEEI 51 KNERARLAAV GERVNQIFTL LGXETALQKG QAGTALATYM LMLERTKSPE 101 VAERALEMAV SLNAFEQAEM IYQKWRQIEP IPGKAQKRAG WLRNVLRERG 151 NQHLDGLEEX LAQADEXQNR RVFLLLAQAA VQQDGLAQKA SKAVRRAALR 201 YEHLPEAAVA DWFSVQXRE KEKAIGALQR LAKLDTEILP PTLMTLRLTA 251 RKYPEILDGF FEQTDTQNLS AVWQEMEIMN LVSLHRLDDA YARLNVLLER 301 NPNADLYIQA AILAANRKEX ASVIDGYAEK AYGRGTGEQR GRAAMTAAMI 351 YADRRDYTKV RQWLKKVSAP EYLFDKGVLA AAAAVELDXG RAALRQIGRV 401 RKLPEQQGRY FTADNLSKIQ MFALSKLPDK REALRGLDKI IEKPPAGSNT 451 ELQAEALVQR SWYDRLGKR KKMISDLERA FRLAPDNAQI MNNLGYSLLS 501 DSKRLDEGFA LLQTAYQINP DDTAVNDSIG WAYYLKXDAE SALPYLRYSF 551 ENDPEPEVAA HLGEVLWALG ERDQAVDVWT QAAHLTGDKK IWRETLKRHG 601 IALPQPSRKP RK* ORF9a and ORF9-1 show 95.3% identity in 614 aa overlap: 10 20 30 40 50 orf9a. pep MLPARFTILSVLAAALLAGQAYAAG--AADAKPPKEVGKVFRKQQRYSEEEIKNERARLA III II : I : I I : I : I : I I I : III Ml I I II II I I II I I I I I I I I I I I I I I I I I I orf9-1 MLPNRFKMLTVLTATLIAGQVSAAGGGAGDMKQPKEVGKVFRKQQRYSEEEIKNERARLA 10 20 30 40 50 60 60 70 80 90 100 110 orf9a . pep AVGERVNQIFTLLGXETALQKGQAGTALATYMLMLERTKSPEVAERALEMAVSLNAFEQA II I I I I II I I I II I II I I I II I II I II II II I I I II I II I I II I I I II I I I II II I II I orf9-1 AVGERVNQIFTLLGGETALQKGQAGTALATYMLMLERTKSPEVAERALEMAVSLNAFEQA 70 80 90 100 110 120 120 130 140 150 160 170 orf9a . pep EMIYQKWRQIEPIPGKAQKRAGWLRNVLRERGNQHLDGLEEXLAQADEXQNRRVFLLLAQ I II I I I I I II I I I I I II I I II I I II I II II I I I I I II I II I I I II II I I II II I I I I I orf9-1 EMIYQKWRQIEPIPGKAQKRAGWLRNVLRERGNQHLDGLEEVLAQADEGQNRRVFLLLAQ 130 140 150 160 170 180 180 190 200 210 220 230 orf9a . pep AAVQQDGLAQKASKAVRRAALRYEHLPEAAVADWFSVQXREKEKAIGALQRLAKLDTEI I II I II I II I II I I I I I I I II : I II I II II I II II II II II I I I I I I I I II I II I I I I I orf9-l AAVQQDGLAQKASKAVRRAALKYEHLPEAAVADWFSVQGREKEKAIGALQRLAKLDTEI 190 200 210 220 230 240 240 250 260 270 280 290 orf9a.pep LPPTLMTLRLTARKYPEILDGFFEQTDTQNLSAVWQEMEIMNLVSLHRLDDAYARLNVLL I I I I II II I I II I I I II I I I II I I I I II I II II II I I II I II I II II I I II I I I II I I I I orf9-1 LPPTLMTLRLTARKYPEILDGFFEQTDTQNLSAVWQEMEIMNLVSLHRLDDAYARLNVLL 250 260 270 280 290 300 300 310 320 330 340 350 orf9a. pep ERNPNADLYIQAAILAANRKEXASVIDGYAEKAYGRGTGEQRGRAAMTAAMIYADRRDYT II I II I II I II II I II II II I I II I II I II II II I I I I II : II I: II II : I I II I I I: orf9-1 ERNPNADLYIQAAILAANRKEGASVIDGYAEKAYGRGTEEQRSRAALTAAMMYADRRDYA 310 320 330 340 350 360 360 370 380 390 400 410 orf9a. pep KVRQWLKKVSAPEYLFDKGVLAAAAAVELDXGRAALRQIGRVRKLPEQQGRYFTADNLSK I II I I I I I I I I I I II I I I II I I I II I II II I I II II II I II I II I I I I II I I I I II II I orf9-1 KVRQWLKKVSAPEYLFDKGVLAAAAAVELDGGRAALRQIGRVRKLPEQQGRYFTADNLSK 370 380 390 400 410 420 420 430 440 450 460 470 orf9a . pep IQMFALSKLPDKREALRGLDKIIEKPPAGSNTELQAEALVQRSWYDRLGKRKKMISDLE I I I: I I I I I I I I I I I I I I I I I I II II I II II I II I I II II I I II I I I I I I II I II I I I I I orf9-1 IQMLALSKLPDKREALRGLDKIIEKPPAGSNTELQAEALVQRSWYDRLGKRKKMISDLE 430 440 450 460 470 480 480 490 500 510 520 530 orf9a . pep RAFRLAPDNAQIMNNLGYΞLLSDSKRLDEGFALLQTAYQINPDDTAVNDSIGWAYYLKXD I I I M I I II II I I I I II I I I 1:11 I II I II II I I I II I I I II I I I I I I I I I I I I I II I I orf9-1 RAFRLAPDNAQIMNNLGYSLLTDSKRLDEGFALLQTAYQINPDDTAVNDSIGWAYYLKGD 490 500 510 520 530 540 540 550 560 570 580 590 orf9a . pep AESALPYLRYSFENDPEPEVAAHLGEVLWALGERDQAVDVWTQAAHLTGDKKIWRETLKR II I II I II I II I I I I I I I II I I I II I II I II II II I II I I I I II I I I M I I II I I I I I I I orf9-1 AESALPYLRYSFENDPEPEVAAHLGEVLWALGERDQAVDVWTQAAHLTGDKKIWRETLKR 550 560 570 580 590 600 600 610 orf9a.pep HGIALPQPSRKPRKX I I I II I I M I I I I I I orf9-l HGIALPQPSRKPRKX 610 Homology with a predicted ORF from N.sonorrhoeae ORF9 shows 82.8% identity over a 163aa overlap with a predicted ORF (ORF9.ng) from N. gonorrhoeae: Orf9 RFKMLTVLTATLIAGQVSAAGGGAGDMKQPKEVGKVFRKQQRYSEEEIKΝERAR 54 M : I: ||: I: I : M I: II | | : | : : || | | I I I : II : : I I I I II I II II II orf9ng MIMLPARFTILSVLAAALLAGQAYAA—GAADVELPKEVGKVLRKHRRYSEEEIKΝERAR 58 orf9 LAAVGERVΝQIFTLLGGETALQKGQAGTALATYMLMLERTKSPEVAERALEMAVSLΝAFE 114 II II II II I :: I II II II I I I I II II I II II I II I II II II II I II I I I I II I II II II I orf9ng LAAVGERVΝRVFTLLGGETALQKGQAGTALATYMLMLERTKSPEVAERALEMAVSLΝAFE 118 orf9 QAEMIYQKWRQIEPIPGKAQKRAGWLRΝVLRERGΝQHLDGREEVLAQADEGQ 166 II II II II II II II II I : I II I II I I I II : I II III III I I : I orf9ng QAEMIYQKWRQIEPIPGEAQKPAGWLRΝVLKEGGΝPHLDRLEEVPAQSDYVHQPMIFLLL 178 The ORF9ng nucleotide sequence <SEQ LD 45> was predicted to encode a protein having including acid sequence <SEQ ID 46>: 1 MIMLPARFTI LSVLAAALLA GQAYAAGAAD VELPKEVGKV LRKHRRYSEE 51 EIKΝERARLA AVGERVΝRVF TLLGGETALQ KGQAGTALAT YMLMLERTKS 101 PEVAERALEM AVSLΝAFEQA EMIYQKWRQI EPIPGEAQKP AGWLRΝVLKE 151 GGΝPHLDRLE EVPAQSDYVH QPMIFLLLVQ AAVQHGGVAQ KPSKAVRPAA 201 YΝYEVLPETA GADAVFCVQG PQYEKAIQSF PPCGRΝPQTE ΝIAPPFΝELF 251 RPTARPISPK LLQRFFRTEP ΝLAKPFRPPG PEMETYQTGF PRPLTRΝΝPT Amino acids 1-28 are a putative leader sequence, and 173-189 are predicted to be a transmembrane domain. Further sequence analysis revealed the complete length ORF9ng DΝA sequence <SEQ LD 47>: 1 ATGTTACCCG CCCGTTTCAC TATTTTATCT GTCCTCGCAG CAGCCCTGCT 51 TGCCGGACAG GCGTATGCTG CCGGCGCGGC GGATGTGGAG CTGCCGAAGG 101 AAGTCGGAAA GGTTTTAAGG AAACATCGGC GTTACAGCGA GGAAGAAATC 151 AAAAACGAAC GCGCACGGCT TGCGGCAGTG GGCGAACGGG TCAACAGGGT 201 GTTTACGCTG TTGGGCGGTG AAACGGCTTT GCAGAAAGGG CAGGCGGGAA 251 CGGCTCTGGC AACCTATATG CTGATGTTGG AACGCACAAA ATCCCCCGAA 301 GTCGCCGAAC GCGCCTTGGA AATGGCCGTG TCGCTGAACG CGTTTGAACA 351 GGCGGAAATG ATTTATCAGA AATGgcggca gatcgagcct ataCcgggtg 401 aggcgcaaaa accgGcgggG tggctgcgga acgtattgaa ggaagggGGa 451 aaTCAGCATC TGGAcgggtt gaaagaggTG CtggcgcaAT cggacgatGT 501 GCAAAAAcgc aggaTATTTT TGCTGCTGGT GCAAGCCGCC GTGCagcagg 551 gTGGGGTGGC TCAAAAAGCA TCGAAAGCGG TTCGCcgtgc GGcgttgaAG 601 TATGAACATC TGCCcgaagc ggcggTTGCC GATGcggTGT TCGGCGTACA 651 GGGACGCGAA AAGGAAAagg caaTCGAAGC TTTGCAGCGT TTGGCGAAGC 701 TCGATACGGA AATATTGCCC CCCACTTTAA TGACGTTGCG TCTGACTGCA 751 CGCAAATATC CCGAAATACT CGACGGCTTT TTCGAGCAGA CAGACACCCA 801 AAACCTTTCG GCCGTCTGGC AGGAAATGGA AATTATGAAT CTGGTTTCCC 851 TGCGTAAGCC GGATGATGCC TATGCGCGTT TGAACGTGCT GTTGGAACAC 901 AACCCGAATG CAAACCTGTA TATTCAGGCG GCGATATTGG CGGCAAACCG 951 AAAAGAAGGT GCGTCCGTTA TCGACGGCTA CGCCGAAAAG GCATACGGCA 1001 GGGGGACGGG GGAACAGCGG GGCagggcgg cAATgacggc GGCGATGATA 1051 TATGCCGACC GCAGGGATTA CGCCAAAGTC AGGCAGTGGT TGAAAAAAGT 1101 GTCCGCGCCG GAATACCTGT TCGACAAAGG CGTGCTGGCG GCTGCGGCGG 1151 CTGCCGAATT GGACGGAGGC CGGGCGGCTT TGCGGCAGAT CGGCAGGGTG 1201 CGGAAACTTC CCGAACAGCA GGGGCGGTAT TTTACGGCAG ACAATTTGTC 1251 CAAAATACAG ATGCTCGCCC TGTCGAAGCT GCCCGACAAA CGGGAAGCCC 1301 TGATCGGGCT GAACAACATC ATCGCCAAAC TTTCGGCGGC GGGAAGCACG 1351 GAACCTTTGG CGGAAGCATT GGCACAGCGT TCCATTATTT ACGaacAGTT 1401 cggCAAACGG GGAAAAATGA TTGCCGACCT tgaAACcgcg CTCAAACTTA 1451 CGCCCGATAA TGCACAAATT ATGAATAATC TGGGCTACAG CCTGCTTTCC 1501 GATTCCAAAC GTTTGGACGA GGGTTTCGCC CTGCTTCAGA CGGCATACCA 1551 AATCAACCCG GACGATACCG CCGTTAACGA CAGCATAGGC TGGGCGTATT 1601 ACCTGAAAGG CGACgcggaA AGCGCGCTGC CGTATCTGcg gtattcgttt 1651 gAAAACGACC CCGAGCCCGA AGTTGCCGCC CATTTGGGCG AAGTGTTGTG 1701 GGCATTGGGC GAACGCGATC AGGCGGTTGA CGTATGGACG CAGGCGGCAC 1751 ACCTTAGGGG AGACAAGAAA ATATGGCGGG AGACGCTCAA ACGCTACGGA 1801 ATCGCCTTGC CCGAGCCTTC CCGAAAACCC CGGAAATAA This encodes a protein having amino acid sequence <SEQ ID 48>: 1 MLPARFTILS VLAAALLAGQ AYAAGAADVE LPKEVGKVLR KHRRYSEEEI 51 KNERARLAAV GERVNRVFTL LGGETALQKG QAGTALATYM LMLERTKSPE 101 VAERALEMAV SLNAFEQAEM IYQKWRQIEP IPGEAQKPAG WLRNVLKEGG 151 NQHLDGLKEV LAQSDDVQKR RIFLLLVQAA VQQGGVAQKA SKAVRRAALK 201 YEHLPEAAVA DAVFGVQGRE KEKAIEALQR LAKLDTEILP PTLMTLRLTA 251 RKYPEILDGF FEQTDTQNLS AVWQEMEIMN LVSLRKPDDA YARLNVLLEH 301 NPNANLYIQA AILAANRKEG ASVIDGYAEK AYGRGTGEQR GRAAMTAAMI 351 YADRRDYAKV RQWLKKVSAP EYLFDKGVLA AAAAAELDGG RAALRQIGRV 401 RKLPEQQGRY FTADNLSKIQ MLALSKLPDK REALIGLNNI IAKLSAAGST 451 EPLAEA AQR SIIYEQFGKR GKMIADLETA LKLTPDNAQI MNNLGYSLLS 501 DSKRLDEGFA LLQTAYQINP DDTAVNDSIG WAYYLKGDAE SALPYLRYSF 551 ENDPEPEVAA HLGEVLWALG ERDQAVDVWT QAAHLRGDKK IWRETLKRYG 601 IALPEPSRKP RK* ORF9ng and ORF9-1 show 88.1% identity in 614 aa overlap: 10 20 30 40 50 60 orf9-1. pep MLPNRFKMLTVLTATLIAGQVSAAGGGAGDMKQPKEVGKVFRKQQRYSEEEIKNERARLA III II : I : II : I : I : I I I : III M : I I 1 I I ! I : II :: I I I I I I I I I M i I II orf9ng-l MLPARFTILSVLAAALLAGQAYAAG--AADVELPKEVGKVLRKHRRYSEEEIKNERARLA 10 20 30 40 50 70 80 90 100 110 120 orf9-1. pep AVGERVNQIFTLLGGETALQKGQAGTALATYMLMLERTKSPEVAERALEMAVSLNAFEQA I I I I I I I:: I I I I I I II I I I I I I II I II I I I I I I II II I II I I I II II I I II II II II II orf9ng-l AVGERVNRVFTLLGGETALQKGQAGTALATYMLMLERTKSPEVAERALEMAVSLNAFEQA 60 70 80 90 100 110 130 140 150 160 170 180 orf9-1. pep EMIYQKWRQIEPIPGKAQKRAGWLRNVLRERGNQHLDGLEEVLAQADEGQNRRVFLLLAQ II II II II I II I II I : I II I I I I I II I : I II I II II I : I II I I : I : I : I I : I I II : I orf9ng-l EMIYQKWRQIEPIPGEAQKPAGWLRNVLKEGGNQHLDGLKEVLAQSDDVQKRRIFLLLVQ 120 130 140 150 160 170 190 200 210 220 230 240 orf9-1. pep AAVQQDGLAQKASKAVRRAALKYEHLPEAAVADWFSVQGREKEKAIGALQRLAKLDTEI I I I II I: I I I II II I II II I I I I I I I I I I I IM I I : II I I I I I II I I I I I I II II II I orf9ng-l AAVQQGGVAQKASKAVRRAALKYEHLPEAAVADAVFGVQGREKEKAIEALQRLAKLDTEI 180 190 200 210 220 230 250 260 270 280 290 300 orf9-1. pep LPPTLMTLRLTARKYPEILDGFFEQTDTQNLSAVWQEMEIMNLVSLHRLDDAYARLNVLL II I II I I I I II I I I I I I I I I I I I II I I I I I II II I I I I II I II I II : : I I I I I I I I I I I orf9ng-l LPPTLMTLRLTARKYPEILDGFFEQTDTQNLSAVWQEMEIMNLVSLRKPDDAYARLNVLL 240 250 260 270 280 290 310 320 330 340 350 360 orf9-1.pep ERNPNADLYIQAAILAANRKEGASVIDGYAEKAYGRGTEEQRSRAALTAAMMYADRRDYA M II I I : I I I II I I II I I II I I I I I I I II I I II II I I I II I : I I I : I I II : I I I II II I orf9ng-l EHNPNANLYIQAAILAANRKEGASVIDGYAEKAYGRGTGEQRGRAAMTAAMIYADRRDYA 300 310 320 330 340 350 370 380 390 400 410 420 orf9-1. pep KVRQWLKKVSAPEYLFDKGVLAAAAAVELDGGRAALRQIGRVRKLPEQQGRYFTADNLSK I II I I I I I I I I I I I I I II I I I I I II M I I I I I II I I I I II I II I II I II I I II I I I I I I I orf9ng-l KVRQWLKKVSAPEYLFDKGVLAAAAAAELDGGRAALRQIGRVRKLPEQQGRYFTADNLSK 360 370 380 390 400 410 430 440 450 460 470 480 orf9-1. pep IQMLALSKLPDKREALRGLDKIIEKPPAGSNTELQAEALVQRSWYDRLGKRKKMISDLE II I I I I II I I II I I I I II : : I I I I : : : I I I I I I : II I : : I : : : I I I II I : I I I orf9ng-l IQMLALSKLPDKREALIGLNNIIAKLSAAGSTEPLAEALAQRSIIYEQFGKRGKMIADLE 420 430 440 450 460 470 490 500 510 520 530 540 orf9-1. pep RAFRLAPDNAQIMNNLGYSLLTDSKRLDEGFALLQTAYQINPDDTAVNDSIGWAYYLKGD M : I : I II I II I I I I II II I I I I I II I II II I II II II I II II II I I I I II II II I I orf9ng-l TALKLTPDNAQIMNNLGYSLLSDSKRLDEGFALLQTAYQINPDDTAVNDSIGWAYYLKGD 480 490 500 510 520 530 550 560 570 580 590 600 orf9-1. pep AESALPYLRYSFENDPEPEVAAHLGEVLWALGERDQAVDVWTQAAHLTGDKKIWRETLKR II I II I I I II II I I I I II II I I II II I I II II I II I II I II II I II I I I I II II I I II I orf9ng-l AESALPYLRYSFENDPEPEVAAHLGEVLWALGERDQAVDVWTQAAHLRGDKKIWRETLKR 540 550 560 570 580 590 610 orf9-1. pep HGIALPQPSRKPRKX : I I II I : II I I I II I orf9ng-l YGIALPEPSRKPRKX 600 610 ition, ORF9ng shows significant homology with a hypothetical protein from P.aeruginosa: sp|P42810|YHE3_PSEAE HYPOTHETICAL 64.8 KD PROTEIN IN HEMM-HEMA INTERGENIC REGION (ORF3) >gi 11072999 Ipir I I S49376 hypothetical protein 3 - Pseudomonas aeruginosa >gi|557259 (X82071) orf3 [Pseudomonas aeruginosa] Length = 576 Score = 128 bits (318), Expect = le-28 Identities = 138/587 (23%), Positives = 228/587 (38%), Gaps = 125/587 (21%) Query: 67 VFTLLGGETALQKGQAGTALATYMLMLERTKSPEVAERALEMAVSLNAFEQAEMIYQKWR 126 +++LL E A Q+ + AL+ Y++ ++T+ P V+ERA +A L A ++A W Sbjct: 53 LYSLLVAELAGQRNRFDIALSNYWQAQKTRDPGVSERAFRIAEYLGADQEALDTSLLWA 112 Query: 127 QIEPIPGEAQKPAG WLRNVLKEGGNQHLDGLKEVLAQSDDVQKRRI 172 + P +AQ+ A ++ VL G+ H D L A++D + + Sbjct: 113 RSAPDNLDAQRAAAIQLARAGRYEESMVYMEKVLNGQGDTHFDFLALSAAETDPDTRAGL 172 Query: 173 FXXXXXXXXXXXXXXXKASKAVRRAALKYEHLPEAAVADAVFGVQGREKEKAIEALQRLA 232 ++ KY + + A+ Q ++A+ L+ + Sbjct: 173 L QSFDHLLKKYPNNGQLLFGKALLLQQDGRPDEALTLLEDNS 214 Query: 233 KLDTEILPPTLMTLRLTARK YPEILDGFFEQTDTQNLSAVWQEMEIMNLVSLRKP 287 E+ P L + L + K P + G E D + + + + LV + Sbjct: 215 ASRHEVAPLLLRSRLLQSMKRSDEALPLLKAGIKEHPDDKRVRLAYARL LVEQNRL 270 Query: 288 DDAYARLNVLLEHNPN ANLYIQAAI 312 DDA A L++ P+ A +Y++ + Sbjct: 271 DDAKAEFAGLVQQFPDDDDDLRFSLALVCLEAQAWDEARIYLEELVERDSHVDAAHFNLG 330 Query: 313 -LAANRKEGASVIDGYAEKAYGRGTGEQRGRAAMTAAMIYADRRDYAKVRQWLKKVSAPE 371 LA +K+ A +D YA+ G G + T ++ A R D A R + P+ Sbjct: 331 RLAEEQKDTARALDEYAQ—VGPGNDFLPAQLRQTDVLLKAGRVDEAAQRLDKARSEQPD 388 Query: 372 YLFDKXXXXXXXXXXXXXXXXXXRQIGRVRKLPEQQGRYFTADNLSKIQMLALSKLPDKR 431 Y A L 1+ ALS + Sbjct: 389 Y AIQLYLIEAEALSNNDQQE 408 Query: 432 EALIGLNNIIAKLSAAGSTEPLAEALAQRSIIYEQFGKRGKMIADLETALKLTPDNAQIM 491 +A + + + E L L RS++ E+ +M DL + PDNA + Sbjct: 409 KAWQAIQEGLKQYP EDL-NLLYTRSMLAEKRNDLAQMEKDLRFVIAREPDNAMAL 462 Query: 492 NNLGYSLLSDSKRLDEGFALLQTAYQINPDDTAVNDSIGWAYYLKGDAESALPYLRYSFE 551 N LGY+L + R E L+ A+++NPDD A+ DS+GW Y +G A YLR + + Sbjct: 463 NALGYTLADRTTRYGEARELILKAHKLNPDDPAILDSMGWINYRQGKLADAERYLRQALQ 522 Query: 552 NDPEPEVAAHLGEVLWALGERDQAVDVWTQAAHLRGDKKIWRETLKR 598 P+ EVAAHLGEVLWA G + A +W + + D + R T+KR Sbjct: 523 RYPDHEVAAHLGEVLWAQGRQGDARAIWREYLDKQPDSDVLRRTIKR 569 gi 12983399 (AE000710) hypothetical protein [Aquifex aeolicus] Length = 545 Score = 81.5 bits (198), Expect = le-14 Identities = 61/198 (30%), Positives = 98/198 (48%), Gaps = 19/198 (9%) Query: 408 GRYFTADNL-SKIQMLALSKLPDKREALIGLNNIIAKLSAAGSTEPLAEALAQ 459 G Y A L K ++LA PDK+E L + +K + + L + Sbjct: 335 GNYEDAKRLIEKAKVLA PDKKEILFLEADYYSKTKQYDKALEILKKLEKDYPNDSR 390 Query: 460 RSIIYEQFGKRGKMIADLETALKLTPDNAQIMNNLGYSLLS—DSKRLDEGFALLQ 513 +I+Y+ G L A++L P+N N LGYSLL +R++E L++ Sbjct: 391 VYFMEAIVYDNLGDIKNAEKALRKAIELDPENPDYYNYLGYSLLLWYGKERVEEAEELIK 450 Query: 514 TAYQINPDDTAVNDSIGWAYYLKGDAESALPYLRYSF-ENDPEPEVAAHLGEVLWALGER 572 A + +P++ A DS+GW YYLKGD E A+ YL + E +P V H+G+VL +G + Sbjct: 451 KALEKDPENPAYIDSMGWVYYLKGDYERAMQYLLKALREAYDDPWNEHVGDVLLKMGYK 510 Query: 573 DQAVDVWTQAAHLRGDKK 590 ++A + + +A L + K Sbjct: 511 EEARNYYERALKLLEEGK 528 Based on this analysis, it is predicted that the proteins from N. meningitidis and N.gonorrhoeae, and their epitopes, could be useful antigens for vaccines or diagnostics, or for raising antibodies. Example 7 The following partial DNA sequence was identified in N. meningitidis <SEQ ID 49>: 1 AACCTCTACG CCGGCCCGCA GACCACATCC GTCATCGCAA ACATCGCCGA 51 CAACCTGCAA CTGGCCAAAG ACTACGGCAA AGTACACTGG TTCGCCTCCC 101 CGCTCTTCTG GCTCCTGAAC CAACTGCACA ACATCATCGG CAACTGGGGC 151 TGGGCGATTA TCGTTTTAAC CATCATCGTC AAAGCCGTAC TGTATCCATT 201 GACCAACGCC TCTTACCGCT CTATGGCGAA AATGCGTGCC GCCGCACCCA 251 AACTGCAAGC CATCAAAGAG AAATACGGCG ACGACCGTAT GGCGCAACAA 301 CAGGCGATGA TGCAGCTTTA CACAGACGAG AAAATCAACC CG-gCTGGGCG 351 GCTGCCTGCC TATGCTGTTG CAAATCCCCG TCTTCATCGG ATTGTATTGG 401 GCATTGTTCG CCTCCGTAGA ATTGCGCCAG GCACCTTGGC TGGGTTGGAT 451 TACCGACCTC AGCCGCGCCG ACCCCTACTA CATCCTGCCC ATCATTATGG 501 CGGCAACGAT GTTCGCCCAA ACTTATCTGA ACCCGCCGCC GAcCGACCCG 551 ATGCagGCGA AAATGATGAA AATCATGCCG TTGGTTTTCT CsGwCrTGTT 601 CTTCTTCTTC CCTGCCGGks TGGTATTGTA CTGGGTAGTC AACAACCTCC 651 TGACCATCGC CCAGCAATGG CACATCAACC GCAGCATCGA AAAACAACGC 701 GCCCAAGGCG AAGTCGTTTC CTAA This corresponds to the amino acid sequence <SEQ ID 50; ORF11>: 1 ..NLYAGPQTTS VIANIADNLQ LAKDYGKVHW FASPLFWLLN QLHNIIGNWG 51 WAIIVLTIIV KAVLYPLTNA SYRSMAKMRA AAPKLQAIKE KYGDDRMAQQ 101 QAMMQLYTDE KINPLGGCLP MLLQIPVFIG LYWALFASVE LRQAPWLGWI 151 TDLSRADPYY ILPIIMAATM FAQTYLNPPP TDPMQAKMMK IMPLVFSXXF 201 FFFPAGXVLY WWNNLLTIA QQWHINRSIE KQRAQGEWS * Further sequence analysis revealed the complete DNA sequence <SEQ LD 51>: 1 ATGGATTTTA AAAGACTCAC GGCGTTTTTC GCCATCGCGC TGGTGATTAT 51 GATCGGCTGG GAAAAGATGT TCCCCACTCC GAAGCCAGTC CCCGCGCCCC 101 AACAGGCAGC ACAACAACAG GCCGTAACCG CTTCCGCCGA AGCCGCGCTC 151 GCGCCCGCAA CGCCGATTAC CGTAACGACC GACACGGTTC AAGCCGTCAT 201 TGATGAAAAA AGCGGCGACC TGCGCCGGCT GACCCTGCTC AAATACAAAG 251 CAACCGGCGA CGAAAATAAA CCGTTCATCC TGTTTGGCGA CGGCAAAGAA 301 TACACCTACG TCGCCCAATC CGAACTTTTG GACGCGCAGG GCAACAACAT 351 TCTAAAAGGC ATCGGCTTTA GCGCACCGAA AAAACAGTAC AGCTTGGAAG 401 GCGACAAAGT TGAAGTCCGC CTGAGCGCGC CTGAAACACG CGGTCTGAAA 451 ATCGACAAAG TTTATACTTT CACCAAAGGC AGCTATCTGG TCAACGTCCG 501 CTTCGACATC GCCAACGGCA GCGGTCAAAC CGCCAACCTG AGCGCGGACT 551 ACCGCATCGT CCGCGACCAC AGCGAACCCG AGGGTCAAGG TTACTTTACC 601 CACTCTTACG TCGGCCCTGT TGTTTATACC CCTGAAGGCA ACTTCCAAAA 651 AGTCAGCTTT TCCGACTTGG ACGACGATGC CAAATCCGGC AAATCCGAGG 701 CCGAATACAT CCGCAAAACC CCGACCGGCT GGCTCGGCAT GATTGAACAC 751 CACTTCATGT CCACCTGGAT TCTCCAACCT AAAGGCAGAC AAAGCGTTTG 801 CGCCGCAGGC GAGTGCAACA TCGACATCAA ACGCCGCAAC GACAAGCTGT 851 ACAGCACCAG CGTCAGCGTG CCTTTAGCCG CCATCCAAAA CGGCGCGAAA 901 GCCGAAGCCT CCATCAACCT CTACGCCGGC CCGCAGACCA CATCCGTCAT 951 CGCAAACATC GCCGACAACC TGCAACTGGC CAAAGACTAC GGCAAAGTAC 1001 ACTGGTTCGC CTCCCCGCTC TTCTGGCTCC TGAACCAACT GCACAACATC 1051 ATCGGCAACT GGGGCTGGGC GATTATCGTT TTAACCATCA TCGTCAAAGC 1101 CGTACTGTAT CCATTGACCA ACGCCTCTTA CCGCTCTATG GCGAAAATGC 1151 GTGCCGCCGC ACCCAAACTG CAAGCCATCA AAGAGAAATA CGGCGACGAC 1201 CGTATGGCGC AACAACAGGC GATGATGCAG CTTTACACAG ACGAGAAAAT 1251 CAACCCGCTG GGCGGCTGCC TGCCTATGCT GTTGCAAATC CCCGTCTTCA 1301 TCGGATTGTA TTGGGCATTG TTCGCCTCCG TAGAATTGCG CCAGGCACCT 1351 TGGCTGGGTT GGATTACCGA CCTCAGCCGC GCCGACCCCT ACTACATCCT 1401 GCCCATCATT ATGGCGGCAA CGATGTTCGC CCAAACTTAT CTGAACCCGC 1451 CGCCGACCGA CCCGATGCAG GCGAAAATGA TGAAAATCAT GCCGTTGGTT 1501 TTCTCCGTCA TGTTCTTCTT CTTCCCTGCC GGTCTGGTAT TGTACTGGGT 1551 AGTCAACAAC CTCCTGACCA TCGCCCAGCA ATGGCACATC AACCGCAGCA 1601 TCGAAAAACA ACGCGCCCAA GGCGAAGTCG TTTCCTAA This corresponds to the amino acid sequence <SEQ ID 52; ORF11-1>: 1 MDFKRLTAFF AIALVIMIGW EKMFPTPKPV PAPQQAAQQQ AVTASAEAAL 51 APATPITVTT DTVQAVIDEK SGDLRRLTLL KYKATGDENK PFILFGDGKE 101 YTYVAQSELL DAQGNNILKG IGFSAPKKQY SLEGDKVEVR LSAPETRGLK 151 IDKVYTFTKG SYLVNVRFDI ANGSGQTANL SADYRIVRDH SEPEGQGYFT 201 HSYVGPWYT PEGNFQKVSF SDLDDDAKSG KSEAEYIRKT PTGWLGMIEH 251 HFMSTWILQP KGRQSVCAAG ECNIDIKRRN DKLYSTSVSV PLAAIQNGAK 301 AEASINLYAG PQTTSVIANI ADNLQLAKDY GKVHWFASPL FWLLNQLHNI 351 IGNWGWAIIV LTIIVKAVLY PLTNASYRSM AKMRAAAPKL QAIKEKYGDD 401 RMAQQQAMMQ LYTDEKINPL GGCLPMLLQI PVFIGLYWAL FASVELRQAP 451 WLGWITDLSR ADPYYILPII MAATMFAQTY LNPPPTDPMQ AKMMKIMPLV 501 FSVMFFFFPA GLVLYWVVNN LLTIAQQWHI NRSIEKQRAQ GEWS* Computer analysis of this amino acid sequence gave the following results: Homology with a 60kDa inner-membrane protein (accession P257541 of Pseudomonas putida ORF11 and the 60kDa protein show 58% aa identity in 229 aa overlap (BLASTp). ORFll 2 LYAGPQTTSVIANIADNLQLAKDYGKVHWFASPLFWLLNQLHNIIGNWGWAIIVLTIIVK 61 LYAGP+ S + ++ L+L DYG + + A P+FWLL +H+++GNWGW+IIVLT+++K 60K 324 LYAGPKIQSKLKELSPGLELTVDYGFLWFIAQPIFWLLQHIHSLLGNWGWSIIVLTMLIK 383 ORFll 62 AVLYPLTNASYRSMAKMRAAAPKLQAIKEKYGDDRXXXXXXXXXLYTDEKINPLGGCLPM 121 + +PL+ ASYRSMA+MRA APKL A+KE++GDDR LY EKINPLGGCLP+ 60K 384 GLFFPLSAASYRSMARMRAVAPKLAALKERFGDDRQKMSQAMMELYKKEKINPLGGCLPI 443 ORFll 122 LLQIPVFIGLYWALFAΞVELRQAPWLGWITDLSRADPYYILPIIMAATMFAQTYLNPPPT 181 L+Q+PVF+ LYW L SVE+RQAPW+ WITDLS DP++ILPIIM ATMF Q LNP P 60K 444 LVQMPVFLALYWVLLESVEMRQAPWILWITDLSIKDPFFILPIIMGATMFIQQRLNPTPP 503 ORFll 182 DPMQAKMMKIMPLVXXXXXXXXPAGXVLYWWNNLLTIAQQWHINRSIE 230 DPMQAK+MK+MP++ PAG VLYWWNN L+I+QQW+I R IE 60K 504 DPMQAKVMKMMPIIFTFFFLWFPAGLVLYWWNNCLSISQQWYITRRIE 552 Homology with a predicted ORF from N.meninsitidis (strain A) ORFll shows 97.9% identity over a 240aa overlap with an ORF (ORF 11a) from strain A of N. meningitidis: 10 20 30 orf11. pep NLYAGPQTTSVIANIADNLQLAKDYGKVHW II II I I I I II I II II I I I II I II I I I II I orf11a IKRRNDKLYSTSVSVPLAAIQNGAKSXASINLYAGPQTTSVIANIADNLQLXKDYGKVHW 280 290 300 310 320 330 40 50 60 70 80 90 orf11.pep FASPLFWLLNQLHNIIGNWGWAIIVLTIIVKAVLYPLTNASYRSMAKMRAAAPKLQAIKE II II I I I I II I II I II I I I I II I I II I II I II II I I II II I II II I II II II II I I II II orf11a FASPLFWLLNQLHNIIGNWGWAIIVLTIIVKAVLYPLTNASYRSMAKMRAAAPKLQAIKE 340 350 360 370 380 390 100 110 120 130 140 150 orf11. pep KYGDDRMAQQQAMMQLYTDEKINPLGGCLPMLLQIPVFIGLYWALFASVELRQAPWLGWI II I I I I I I II II I I II I I I I II I II I I II II II I I I I I I II I I II I I I II I I II I I I I I I orf11a KYGDDRMAQQQAMMQLYTDEKINPLGGCLPMLLQIPVFIGLYWALFASVELRQAPWLGWI 400 410 420 430 440 450 160 170 180 190 200 210 orf11. pep TDLSRADPYYILPIIMAATMFAQTYLNPPPTDPMQAKMMKIMPLVFSXXFFFFPAGXVLY I II II I I II I I I II II I I I I II I II I I I I II II I I I I I I I I I I II I I I I I II I I III orf11a TDLSRADPYYILPIIMAATMFAQTYLNPPPTDPMQAKMMKIMPLVXSXXFFXFPAGLVLY 460 470 480 490 500 510 220 230 240 orf11.pep WVVNNLLTIAQQWHINRSIEKQRAQGEWSX I I : II II II II I II I II I I I II I II I I I I II orf11a WVINNLLTIAQQWHINRSIEKQRAQGEVVSX 520 530 540 The complete length ORF1 la nucleotide sequence <SEQ ID 53> is: 1 ANGGATTTTA AAAGACTCAC NGNGTTTTTC GCCATCGCAC TGGTGATTAT 51 GATCGGATNG NAAANGATGT TCCCCACTCC GAAGCCCGTC CCCGCGCCCC 101 AACAGACGGC ACAACAACAG GCCGTAANCG CTTCCGCCGA AGCCGCGCTC 151 GCGCCCGNAN CGCCGATTAC CGTAACGACC GACACGGTTC AAGCCGTCAT 201 TGATGAAAAA AGCGGCGACC TGCGCCGGCT GACCCTGCTC AAATACAAAG 251 CAACCGGCGA CNAAAATAAA CCGTTCATCC TGTTTGGCGA CGGCAAANAA 301 TACACCTACN TCGCCCANTC CGAACTTTTG GACGCGCAGG GCAACAACAT 351 TCTAAAAGGC ATCGGCTTTA GCGCACCGAA AAAACAGTAC AGCTTGGAAG 401 GCGACAAAGT TGAAGTCCGC CTGAGCGCAC CTGAAACACG CGGTCTGAAA 451 ATCGACAAAG TTTATACTTT CACCAAAGGC AGCTATCTGG TCAACGTCCG 501 CTTCGACATC GCCAACGGCA GCGGTCAAAC CGCCAACCTG AGCGCGGACT 551 ACCGCATCGT CCGCGACCAC AGCGAACCCG AGGGTCAAGG CTACTTTACC 601 CACTCTTACG TCGGCCCTGT TGTTTATACC CCTGAAGGCA ACTTCCAAAA 651 AGTCAGCTTC TCCGACTTGG ACGACGATGC CAANTCCGGN AAATCCGAGG 701 CCGAATACAT CCGCAAAACC CNGACCGGCT GGCTCGGCAT GATTGAACAC 751 CACTTCATGT CCACCTGGAT CCTCCAACCC AAAGGCGGAC AAAGCGTTTG 801 CGCCGCTGGC GACTGCNGTA TNGACATCAA ACGCCGCAAC GACAAGCTGT 851 ACAGCACCAG CGTCAGCGTG CCTTTAGCCG CTATCCAAAA CGGTGCGAAA 901 TCCNAAGCCT CCATCAACCT CTACGCCGGC CCACAGACCA CATCNGTTAT 951 CGCAAACATC GCCGACAACC TGCAACTGGN CAAAGACTAC GGCAAAGTAC 1001 ACTGGTTCGC CTCCCCCCTC TTTTGGCTTT TGAACCAACT GCACAACATC 1051 ATCGGCAACT GGGGCTGGGC GATTATCGTT TTAACCATCA TCGTCAAAGC 1101 CGTACTGTAT CCATTGACCA ACGCCTCTTA CCGTTCGATG GCGAAAATGC 1151 GTGCCGCCGC GCCCAAACTG CAAGCCATCA AAGAGAAATA CGGCGACGAC 1201 CGTATGGCGC AGCAACAAGC CATGATGCAG CTTTACACAG ACGAGAAAAT 1251 CAACCCGCTG GGCGGCTGCC TGCCTATGCT GTTGCAAATC CCCGTCTTCA 1301 TCGGATTGTA TTGGGCATTG TTCGCCTCCG TAGAATTGCG CCAGGCACCT 1351 TGGCTGGGTT GGATTACCGA CCTCAGCCGC GCCGACCCNT ACTACATCCT 1401 GCCCATCATT ATGGCGGCAA CGATGTTCGC CCAAACCTAT CTGAACCCGC 1451 CGCCGACCGA CCCGATGCAG GCGAAAATGA TGAAAATCAT GCCTTTGGTT 1501 NTNTCNNNNA NGTTCTTCNN CTTCCCTGCC GGTCTGGTAT TGTACTGGGT 1551 GATCAACAAC CTCCTGACCA TCGCCCAGCA ATGGCACATC AACCGCAGCA 1601 TCGAAAAACA ACGCGCCCAA GGCGAAGTCG TTTCCTAA This encodes a protein having amino acid sequence <SEQ LD 54>: 1 XDFKRLTXFF AIALVIMIGX XXMFPTPKPV PAPQQTAQQQ AVXASAEAAL 51 APXXPITVTT DTVQAVIDEK SGDLRRLTLL KYKATGDXNK PFILFGDGKX 101 YTYXAXSELL DAQGNNILKG IGFSAPKKQY SLEGDKVEVR LSAPETRGLK 151 IDKVYTFTKG SYLVNVRFDI ANGSGQTANL SADYRIVRDH SEPEGQGYFT 201 HSYVGPWYT PEGNFQKVSF SDLDDDAXSG KSEAEYIRKT XTGWLGMIEH 251 HFMSTWILQP KGGQSVCAAG DCXXDIKRRN DKLYSTSVSV PLAAIQNGAK 301 SXASINLYAG PQTTSVIANI ADNLQLXKDY GKVHWFASPL FWLLNQLHNI 351 IGNWGWAIIV LTIIVKAVLY PLTNASYRSM AKMRAAAPKL QAIKEKYGDD 401 RMAQQQAMMQ LYTDEKINPL GGCLPMLLQI PVFIGLYWAL FASVELRQAP 451 WLGWITDLSR ADPYYILPII MAATMFAQTY LNPPPTDPMQ AKMMKIMPLV 501 XSXXFFXFPA GLVLYWVINN LLTIAQQWHI NRSIEKQRAQ GEWS* ORF1 la and ORF11-1 show 95.2% identity in 544 aa overlap: 10 20 30 40 50 60 orf11a . pep XDFKRLTXFFAIALVIMIGXXXMFPTPKPVPAPQQTAQQQAVXASAEAALAPXXPITVTT MMM I II I I I II I I I I I II II I II II I I : I II I II : I I II II II I : I I I I I I orf11-1 MDFKRLTAFFAIALVIMIGWEKMFPTPKPVPAPQQAAQQQAVTASAEAALAPATPITVTT 10 20 30 40 50 60 70 80 90 100 110 120 orflla.pep DTVQAVIDEKSGDLRRLTLLKYKATGDXNKPFILFGDGKXYTYXAXSELLDAQGNNILKG II II II II I II I I II I I I II I II I I I I II II II II II I III I I II I II I I I I I I II orf11-1 DTVQAVIDEKSGDLRRLTLLKYKATGDENKPFILFGDGKEYTYVAQSELLDAQGNNILKG 70 80 90 100 110 120 130 140 150 160 170 180 orflla.pep IGFSAPKKQYSLEGDKVEVRLSAPETRGLKIDKVYTFTKGSYLVNVRFDIANGSGQTANL II II II I II II II II II II II I II I II II I II I II II I II II II II II I II I I I I I I II I orfll-1 IGFSAPKKQYSLEGDKVEVRLSAPETRGLKIDKVYTFTKGSYLVNVRFDIANGSGQTANL 130 140 150 160 170 180 190 200 210 220 230 240 orf11a . pep SADYRIVRDHSEPEGQGYFTHSYVGPWYTPEGNFQKVSFSDLDDDAXSGKSEAEYIRKT I I II II II I II I I II II II I II I I II I I I II II II II II II I II II I II II I I I I I I I I orf11-1 SADYRIVRDHSEPEGQGYFTHSYVGPWYTPEGNFQKVSFSDLDDDAKSGKSEAEYIRKT 190 200 210 220 230 240 250 260 270 280 290 300 orflla.pep XTGWLGMIEHHFMSTWILQPKGGQSVCAAGDCXXDIKRRNDKLYSTSVSVPLAAIQNGAK I II II I I I II II II II I I I I I I I I II IM I I- 1 II II I II I I II II II I I I I I I I I.I orfll-1 PTGWLGMIEHHFMSTWILQPKGRQSVCAAGECNIDIKRRNDKLYSTSVSVPLAAIQNGAK 250 260 270 280 290 300 310 320 330 340 350 360 orf11a . pep SXASINLYAGPQTTSVIANIADNLQLXKDYGKVHWFASPLFWLLNQLHNIIGNWGWAIIV : I II II I I I II II I I I I II I II I I I II I II I I I II I I I II II II I I II II I I I I I I I I orf11-1 AEASINLYAGPQTTSVIANIADNLQLAKDYGKVHWFASPLFWLLNQLHNIIGNWGWAIIV 310 320 330 340 350 360 370 380 390 400 410 420 orflla.pep LTIIVKAVLYPLTNASYRSMAKMRAAAPKLQAIKEKYGDDRMAQQQAMMQLYTDEKINPL I II I I I II I II II I I I I I II I II I I II II I II I II II I I II I I I II I II I I II I I II I I I orf11-1 LTIIVKAVLYPLTNASYRSMAKMRAAAPKLQAIKEKYGDDRMAQQQAMMQLYTDEKINPL 370 380 390 400 410 420 430 440 450 460 470 480 orflla.pep GGCLPMLLQIPVFIGLYWALFASVELRQAPWLGWITDLSRADPYYILPIIMAATMFAQTY II I II I II II I I I II I I I II I II I I II II I I I I II II I I I II II II I II I I II I I II I I I orfll-1 GGCLPMLLQIPVFIGLYWALFASVELRQAPWLGWITDLSRADPYYILPIIMAATMFAQTY 430 440 450 460 470 480 490 500 510 520 530 540 orflla.pep LNPPPTDPMQAKMMKIMPLVXSXXFFXFPAGLVLYWVINNLLTIAQQWHINRSIEKQRAQ II I II II II I II I II II II I I II I II II I II I I : II II I I II I I II II I I I I I I II orf11-1 LNPPPTDPMQAKMMKIMPLVFSVMFFFFPAGLVLYWWNNLLTIAQQWHINRSIEKQRAQ 490 500 510 520 530 540 orflla.pep GEWSX I I I I I I orfll-1 GEWSX Homology with a predicted ORF from N.sonorrhoeae ORFl l shows 96.3% identity over a 240aa overlap with a predicted ORF (ORFl l. ng) from N gonorrhoeae: Orfll NLYAGPQTTSVIANIADNLQLAKDYGKVHWFASPLFWLLNQLHNIIGNWGWAIIVLT 57 I I II II I I I II I II II I II I II I I II II II II II I I II II II II II I II II I I: I I I orfling MAVNLYAGPQTTSVIANIADNLQLAKDYGKVHWFASPLFWLLNQLHNIIGNWGWAIWLT 60 orfll IIVKAVLYPLTNASYRSMAKMRAAAPKLQAIKEKYGDDRMAQQQAMMQLYTDEKINPLGG 117 I II I II II I II II II II I I I I I I II I : I I : II II I II II I II I I II II I : I IIIMI orfllng IIVKAVLYPLTNASYRSMAKMRAAAPELQTIKEKYGDDRMAQQQAMMQLFEDEEINPLGG 120 orfll CLPMLLQIPVFIGLYWALFASVELRQAPWLGWITDLSRADPYYILPIIMAATMFAQTYLN 177 I I II I II I II II I I I I II I II I II II I I I II I I I II II I II I II I II I II II II II I I I I orfllng CLPMLLQIPVFIGLYWALFASVELRQAPWLGWITDLSRADPYYILPIIMAATMFAQTYLN 180 orfll PPPTDPMQAKMMKIMPLVFSXXFFFFPAGXVLYWWNNLLTIAQQWHINRSIEKQRAQGE 237 II I I I I I I II I I II I II II I I II II I I II II I II II I II I I II M I I I II II I I I I I orfllng PPPTDPMQAKMMKIMPLVFSVMFFFFPAGLVLYWWNNLLTIAQQWHINRSIEKQRAQGE 240 orfll WS 240 I I I orfllng WS 243 An ORFl lng nucleotide sequence <SEQ LD 55> was predicted to encode a protein having amino acid sequence <SEQ LD 56>: 1 MAVNLYAGPQ TTSVIANIAD NLQLAKDYGK VHWFASPLFW LLNQLHNIIG 51 NWGWAIWLT IIVKAVLYPL TNASYRSMAK MRAAAPELQT IKEKYGDDRM 101 AQQQAMMQLF EDEEINPLGG CLPMLLQIPV FIGLYWALFA SVELRQAPWL 151 GWITDLSRAD PYYILPIIMA ATMFAQTYLN PPPTDPMQAK MMKIMPLVFS 201 VMFFFFPAGL VLYWWNNLL TIAQQWHINR SIEKQRAQGE WS* Further sequence .analysis revealed the complete gonococcal DNA sequence <SEQ LD 57> to be: 1 ATGGATTTTA AAAGACTCAC GGCGTTTTTC GCCATCGCGC TGGTGATTAT 51 GATCGGCTGG GAAAAAATGT TCCCCACCCC GAAACCCGTC CCCGCGCCCC 101 AACAGGCGGC ACAAAAACAG GCAGCAACCG CTTCCGCCGA AGCCGCGCTC 151 GCGCCCGCAA CGCCGATTAC CGTAACGACC GACACGGTTC AAGCCGTTAT 201 TGATGAAAAA AGTGGCGACC TGCGCCGGCT GACCCTGCTC AAATACAAAG 251 CAACCGGCGA CGAAAACAAA CCGTTCGTCC TGTTTGGCGA CGGCAAAGAA 301 TACACCTACG TCGCCCAATC CGAACTTTTG GACGCGCAGG GCAACAACAT 351 TCTGAAAGGC ATCGGCTTTA GCGCACCGAA AAAACAGTAC ACCCTCAACG 401 GCGACACAGT CGAAGTCCGC CTGAGCGCGC CCGAAACCAA CGGACTGAAA 451 ATCGACAAAG TCTATACCTT TACCAAAGAC AGCTATCTGG TCAACGTCCG 501 CTTCGACATC GCCAACGGCA GCGGTCAAAC CGCCAACCTG AGCGCGGACT 551 ACCGCATCGT CCGCGACCAC AGCGAACCCG AGGGTCAAGG CTACTTTACC 601 CACTCTTACG TCGGCCCTGT TGTTTATACC CCTGAAGGCA ACTTCCAAAA 651 AGTCAGCTTC TCCgacTTgg acgACGATGC gaaaTccggc aaATccgagg 701 ccgaatacaT CCGCAAAACC ccgaccggtt ggctcggcat gattgaacac 751 cacttcatgt ccacctggat cctccAAcct aaaggcggcc aaaacgtttg 801 cgcccaggga gactgccgta tcgacattaa aCgccgcaac gacaagctgt 851 acagcgcaag cgtcagcgtg cctttaaccg ctatcccaac ccgggggcca 901 aaaccgaaaa tggcggTCAA CCTGTATGCC GGTCCGCAAA CCACATCCGT 951 TATCGCAAAC ATCGCcgacA ACCTGCAACT GGCAAAAGAC TACGGTAAAG 1001 TACACTGGTT CGCATCGCCG CTCTTCTGGC TCCTGAACCA ACTGCACAAC 1051 ATTATCGGCA ACTGGGGCTG GGCAATCGTC GTTTTGACCA TCATCGTCAA 1101 AGCCGTACTG TATCCATTGA CCAACGcctc ctACCGTTCG ATGGCGAAAA 1151 TGCGTGccgc cgcacCcaaA CTGCAGACCA TCAAAGAAAA ATAcgGCGAC 1201 GACCGTATGG CGCAACAGCA AGCGATGATG CAGCTTTACA AAgacgAGAA 1251 AATCAACCCG CTGGGCGGCT GTctgcctat gctgttgCAA ATCCCCGTCT 1301 TCATCGGCTT GTACTGGGCA TTGTTCGCCT CCGTAGAATT GCGCCAGGCA 1351 CCTTGGCTGG GCTGGATTAC CGACCTCAGC CGCGCCGACC CCTACTACAT 1401 CCTGCCCATC ATTATGGCGG CAACGATGTT CGCCCAAACC TATCTGAACC 1451 CGCCGCCGAC CGACCCGATG CAGGCGAAAA TGATGAAAAT CATGCCGTTG 1501 GTTTTCTCCG TCATGTTCTT CTTCTTCCCT GCCGGTTTGG TTCTCTACTG 1551 GGTGGTCAAC AACCTCCTGA CCATCGCCCA GCAGTGGCAC ATCAACCGCA 1601 GCATCGAAAA ACAACGCGCC CAAGGCGAAG TCGTTTCCTA A This encodes a protein having amino acid sequence <SEQ LD 58; ORFl lng-l>: 1 MDFKRLTAFF AIALVIMIGW EKMFPTPKPV PAPQQAAQKQ AATASAEAAL 51 APATPITVTT DTVQAVIDEK SGDLRRLTLL KYKATGDENK PFVLFGDGKE 101 YTYVAQSELL DAQGNNILKG IGFSAPKKQY TLNGDTVEVR LSAPETNGLK 151 IDKVYTFTKD SYLVNVRFDI ANGSGQTANL SADYRIVRDH SEPEGQGYFT 201 HSYVGPWYT PEGNFQKVSF SDLDDDAKSG KSEAEYIRKT PTGWLGMIEH 251 HFMSTWILQP KGGQNVCAQG DCRIDIKRRN DKLYSASVSV PLTAIPTRGP 301 KPKMAVNLYA GPQTTSVIAN IADNLQLAKD YGKVHWFASP LFWLLNQLHN 351 IIGNWGWAIV VLTIIVKAVL YPLTNASYRS MAKMRAAAPK LQTIKEKYGD 401 DRMAQQQAMM QLYKDEKINP LGGCLPMLLQ IPVFIGLYWA LFASVELRQA 451 PWLGWITDLS RADPYYILPI IMAATMFAQT YLNPPPTDPM QAKMMKIMPL 501 VFSVMFFFFP AGLVLYWWN NLLTIAQQWH INRSIEKQRA QGEWS* ORFl lng-1 and ORFl 1-1 shown 95.1% identity in 546 aa overlap: 10 20 30 40 50 60 orfling-1.pep MDFKRLTAFFAIALVIMIGWEKMFPTPKPVPAPQQAAQKQAATASAEAALAPATPITVTT I I II I I II II I I II I I I II I I I I I II I I II I II II I IM IM I II I II II II I I I I I I II orfll-1 MDFKRLTAFFAIALVIMIGWEKMFPTPKPVPAPQQAAQQQAVTASAEAALAPATPITVTT 10 20 30 40 50 60 70 80 90 100 110 120 orfling-1.pep DTVQAVIDEKSGDLRRLTLLKYKATGDENKPFVLFGDGKEYTYVAQSELLDAQGNNILKG I II II II II II I I I II II MM II I II II II I: I I I I II II I II I I I I II I I I II I I II I orfll-1 DTVQAVIDEKSGDLRRLTLLKYKATGDENKPFILFGDGKEYTYVAQSELLDAQGNNILKG 70 80 90 100 110 120 130 140 150 160 170 180 orfling-1.pep IGFSAPKKQYTLNGDTVEVRLSAPETNGLKIDKVYTFTKDSYLVNVRFDIANGSGQTANL I II I II I II I : I : I I II I I II I II I I II II I II II I I I II II I II I I I II I I I I I I I orfll-1 IGFSAPKKQYSLEGDKVEVRLSAPETRGLKIDKVYTFTKGSYLVNVRFDIANGSGQTANL 130 140 150 160 170 180 190 200 210 220 230 240 orfling-1.pep SADYRIVRDHSEPEGQGYFTHSYVGPWYTPEGNFQKVSFSDLDDDAKSGKSEAEYIRKT I II II II II I I I II I I I I II I II I I II II I I I II I I I II II I I II I II I I II I I I I I I I I orfll-1 SADYRIVRDHSEPEGQGYFTHSYVGPWYTPEGNFQKVSFSDLDDDAKSGKSEAEYIRKT 190 200 210 220 230 240 250 260 270 280 290 300 orfling-1.pep PTGWLGMIEHHFMSTWILQPKGGQNVCAQGDCRIDIKRRNDKLYSASVSVPLTAIPTRGP I II II II II I I I II I I I I II II ill l I I II II II I I II : I II I I I : II : I orfll-1 PTGWLGMIEHHFMSTWILQPKGRQSVCAAGECNIDIKRRNDKLYSTSVSVPLAAIQN-GA 250 260 270 280 290 310 320 330 340 350 360 orfllng-1. pep KPKMAVNLYAGPQTTSVIANIADNLQLAKDYGKVHWFASPLFWLLNQLHNIIGNWGWAIV I : :: I II I II II II I I II I I II I II I II II I II II I II I II I I II II II I I II I I IM orf11-1 KAEASINLYAGPQTTSVIANIADNLQLAKDYGKVHWFASPLFWLLNQLHNIIGNWGWAII 300 310 320 330 340 350 370 380 390 400 410 420 orfllng-l.pep VLTIIVKAVLYPLTNASYRSMAKMRAAAPKLQTIKEKYGDDRMAQQQAMMQLYKDEKINP I II I II I II I II II I I I I I I I I II I I I I II IM I I II I II I II II I I I I II II I I I II I orf11-1 VLTIIVKAVLYPLTNASYRSMAKMRAAAPKLQAIKEKYGDDRMAQQQAMMQLYTDEKINP 360 370 380 390 400 410 430 440 450 460 470 480 orfllng-l.pep LGGCLPMLLQIPVFIGLYWALFASVELRQAPWLGWITDLSRADPYYILPIIMAATMFAQT II II I I II I II II I I I I I II I I I I I II II I II I I I I I II II I I II I II II I II I I III I I orf11-1 LGGCLPMLLQIPVFIGLYWALFASVELRQAPWLGWITDLSRADPYYILPIIMAATMFAQT 420 430 440 450 460 470 490 500 510 520 530 540 orfllng-1. pep YLNPPPTDPMQAKMMKIMPLVFSVMFFFFPAGLVLYWWNNLLTIAQQWHINRSIEKQRA I I I II I I I II I I I II I I I I I I II I I II II II I II II II I II I II II II II I II II I II I I orf11-1 YLNPPPTDPMQAKMMKIMPLVFSVMFFFFPAGLVLYWWNNLLTIAQQWHINRSIEKQRA 480 490 500 510 520 530 orfllng-l . pep QGEWSX I I I I I I I orfll-1 QGEWSX 540 In addition, ORFl lng-1 shows significant homology with an inner-membrane protein from the database (accession number p25754): ID 60IM_PSEPU STANDARD; PRT; 560 AA. AC P25754; DT 01-MAY-1992 (REL. 22, CREATED) DT 01-MAY-1992 (REL. 22, LAST SEQUENCE UPDATE) DT 01-NOV-1995 (REL. 32, LAST ANNOTATION UPDATE) DE 60 KD INNER-MEMBRANE PROTEIN. . . . SCORES Initl: 1074 Initn: 1293 Opt: 1103 Smith-Waterman score: 1406; 41.5% identity in 574 aa overlap 10 20 30 40 orfllng-l.pep MDFKR LTAFFAIALVIMIGW EKMFPT PKPVPAPQQAAQKQ IMM :: I : ::: I ::: I : : II I II I ::: I : : p25754 MDIKRTILIAALAVVSYVMVLKWNDDYGQAALPTQNTAASTVAPGLPDGVPAGNNGASAD 10 20 30 40 50 60 50 60 70 80 90 orfllng-l.pep AATASAEAALAPATPIT VTTDTVQAVIDEKSGDLRRLTLLKYKATGDE-NKPF : : I: I I :: I : I :: | || ::: : I I : || : : | : I II M i ll p25754 VPSANAESSPAELAPVALSKDLIRVKTDVLELAIDPVGGDIVQLNLPKYPRRQDHPNIPF 70 80 90 100 110 120 100 110 120 130 140 orfllng-l.pep VLFGDGKEYTYVAQSELLDAQGNNILKGIG FSAPKKQYTL-NGD TVEVRLSAPE I I : I I : I : II I I : : I : : : I : : I : I : I I : I : : I : : : : I p25754 QLFDNGGERVYLAQSGLTGTDGPDA-RASGRPLYAAEQKSYQLADGQEQLWDLKFS 130 140 150 160 170 150 160 170 180 190 200 orf llng-1 . pep TNGLKIDKVYTFTKDSYLVNVRFDIANGSGQTANLSADYRIVRDHS-EPEGQGYF-THSY I I : : I : : I : I : I I : I I I I I : I : : : I I I : I : : I : I p25754 DNGVNYIKRFSFKRGEYDLNVSYLIDNQSGQAWNGNMFAQLKRDASGDPSSSTATGTATY 180 190 200 210 220 230 210 220 230 240 250 260 orf ling- 1 . pep VGPWYTPEGNFQKVSFSDLDDDAKSGKSEAEYIRKTPTGWLGMIEHHFMSTWILQPKGG : I : : : I : : | | | : : | : | I : : : | : : | | : : : : | : | : : : | | | : p25754 LGAALWTASEPYKKVSMKDID KGSLKE NVSGGWVAWLQHYFVTAWI-PAKSD 240 250 260 270 280 270 280 290 300 310 320 orfling-1. pep QNVCAQGDCRIDIKRRNDKLYSASVSVPLTAIPTRGPKPKMAVNLYAGPQTTSVIANIAD : 1 I :: :: :: I : : | : : : | : I I : : : II I I I : I : :: : p25754 NNV VQTRKDSQGNYIIGYTGPVISVPA-GGKVETSALLYAGPKIQSKLKELSP 290 300 310 320 330 330 340 350 360 370 380 orfllng-l.pep NLQLAKDYGKVHWF-ASPLFWLLNQLHNIIGNWGWAIWLTIIVKAVLYPLTNASYRSMA : I: I : III : II I : I : I II I : : : I : : : I I II I : I : I I I : : : I : : : : II : I I I I II I p25754 GLELTVDYGFL-WFIAQPIFWLLQHIHSLLGNWGWSIIVLTMLIKGLFFPLSAASYRSMA 340 350 360 370 380 390 390 400 410 420 430 440 orfllng-l.pep KMRAAAPKLQTIKEKYGDDRMAQQQAMMQLYKDEKINPLGGCLPMLLQIPVFIGLYWALF : I I I : I I I I : : II :: I M I : :: I I I I : II I I II II I I II I I : I : I : II I : : I I I : I : p25754 RMRAVAPKLAALKERFGDDRQKMSQAMMELYKKEKINPLGGCLPILVQMPVFLALYWVLL 400 410 420 430 440 450 450 460 470 480 490 500 orfling-1. pep ASVELRQAPWLGWITDLSRADPYYILPIIMAATMFAQTYLNPPPTDPMQAKMMKIMPLVF I II : I II I I : I I I II I I I:: I II II M II I I I III I II I I I I : I I : I I : : I p25754 ESVEMRQAPWILWITDLSIKDPFFILPIIMGATMFIQQRLNPTPPDPMQAKVMKMMPIIF 460 470 480 490 500 510 510 520 530 540 orfling-1.pep SVMFFFFPAGLVLYWWNNLLTIAQQWHINRSIEKQRAQGEWSX : : I:: I II I II II II I II M I I M I : I II p25754 TFFFLWFPAGLVLYWWNNCLSISQQWYITRRIEAATKKAAA 520 530 540 550 560 Based on this analysis, including the homology to an inner-membrane protein from P. putida and the predicted transmembrane domains (seen in both the meningococcal and gonoccal proteins), it is predicted that the proteins from N. meningitidis .and N.gonorrhoeae, .and their epitopes, could be useful antigens for vaccines or diagnostics, or for raising antibodies. Example 8 The following partial DNA sequence was identified in N. meningitidis <SEQ ID 59>: 1 .. GCCGTCTTAA TCATCGAATT ATTGACGGGA ACGGTTTATC TTTTGGTTGT 51 NAGCGCGGCT TTGGCGGGTT CGGGCATTGC TTACGGGCTG ACCGGCAGTA 101 CGCCTGCCGC CGTCTTGACC GNCGCTCTGC TTTCCGCGCT GGGTATTTNG 151 TTCGTACACG CCAAAACCGC CGTTAGAAAA GTTGAAACGG ATTCATATCA 201 GGATTTGGAT GCCGGACAAT ATGTCGAAAT CCTCCGNCAC ACAGGCGGCA 251 ACCGTTACGA AGTT.TTTAT CGCGGTACG. ACTGGCAGGC TCAAAATACG 301 GGGCAAGAAG AGCTTGAACC AGGAACTCGC GCCCTCATTG TCCGCAAGGA 351 AGGCAACCTT CTTATTATCA CACACCCTTA A This corresponds to the amino acid sequence <SEQ ID 60; ORF13>: 1 ..AVLIIELLTG TVYLL SAA LAGSGIAYGL TGSTPAAVLT XALLSALGIX 51 FVHAKTAVRK VETDSYQDLD AGQYVEILRH TGGNRYEVXY RGTXWQAQNT 101 GQEELEPGTR ALIVRKEGNL LIITHP* Further sequence analysis elaborated the DNA sequence slightly <SEQ ID 61>: 1 .. GCCGTCTTAA TCATCGAATT ATTGACGGGA ACGGTTTATC TTTTGGTTGT 51 nAGCGCGGCT TTGGCGGGTT CGGGCATTGC TTACGGGCTG ACCGGCAGTA 101 CGCCTGCCGC CGTCTTGACC GnCGCTCTGC TTTCCGCGCT GGGTATTTnG 151 TTCGTACACG CCAAAACCGC CGTTAGAAAA GTTGAAACGG ATTCATATCA 201 GGATTTGGAT GCCGGACAAT ATGTCGAAAT CCTCCGACAC ACAGGCGGCA 251 ACCGTTACGA AGTTTTtTAT CGCGGTACGc ACTGGCAGGC TCAAAATACG 301 GGGCAAGAAG AGCTTGAACC AGGAACTCGC GCCCTCATTG TCCGCAAGGA 351 AGGCAACCTT CTTATTATCA CACACCCTTA A This corresponds to the amino acid sequence <SEQ ID 62; ORF13-l>: 1 ..AVLIIELLTG TVYLLWSAA LAGSGIAYGL TGSTPAAVLT XALLSALGIX 51 FVHAKTAVRK VETDSYQDLD AGQYVEILRH TGGNRYEV Y RGTHWQAQNT 101 GQEELEPGTR ALIVRKEGNL LIITHP* Computer analysis of this amino acid sequence gave the following results: Homology with a predicted ORF from N.meninsitidis (strain Al ORF13 shows 92.9% identity over a 126aa overlap with an ORF (ORF13a) from strain A of N. meningitidis: 10 20 30 40 50 orf13.pep AVLIIELLTGTVYLLWSAALAGSGIAYGLTGSTPAAVLTXALLSALGIXF I I II I I II II I I II I I I I II II I I II I I II I I I I I II I I I I II II I I I I orfl3a MTVWFVAAVAVLIIELLTGTVYLLWSAALAGSGIAYGLTGSTPAAVLTAALLSALGIWF 10 20 30 40 50 60 60 70 80 90 100 110 orf13.pep VHAKTAVRKVETDSYQDLDAGQYVEILRHTGGNRYEVXYRGTXWQAQNTGQEELEPGTRA II II I I I I I II I I II I I I I II I : I II I I : I II I I II I I I I I I I I I II I II II I I I I I orf13a VHAKTAVGKVETDSYQDLDAGQYAEILRHAGGNRYEVFYRGTHWQAQNTGQEELEPGTRA 70 80 90 100 110 120 120 orf13.pep LIVRKEGNLLIITHPX I I I I II I I I II I :: M orf 13a LIVRKEGNLLIIAKPX 130 The complete length ORF13a nucleotide sequence <SEQ ID 63> is: 1 ATGACTGTAT GGTTTGTTGC CGCTGTTGCC GTCTTAATCA TCGAATTATT 51 GACGGGAACG GTTTATCTTT TGGTTGTCAG CGCGGCTTTG GCGGGTTCGG 101 GCATTGCTTA CGGGCTGACC GGCAGCACGC CTGCCGCCGT CTTGACCGCC 151 GCTCTGCTTT CCGCGCTGGG TATTTGGTTC GTACACGCCA AAACCGCCGT 201 GGGAAAAGTT GAAACGGATT CATATCAGGA TTTGGATGCC GGGCAATATG 251 CCGAAATCCT CCGGCACGCA GGCGGCAACC GTTACGAAGT TTTTTATCGC 301 GGTACGCACT GGCAGGCTCA AAATACGGGG CAAGAAGAGC TTGAACCAGG 351 AACGCGCGCC CTAATCGTCC GCAAGGAAGG CAACCTTCTT ATCATCGCAA 401 AACCTTAA This encodes a protein having amino acid sequence <SEQ LD 64>: 1 MTVWFVAAVA VLIIELLTGT VYLLWSAAL AGSGIAYGLT GSTPAAVLTA 51 ALLSALGIWF VHAKTAVGKV ETDSYQDLDA GQYAEILRHA GGNRYEVFYR 101 GTHWQAQNTG QEELEPGTRA LIVRKEGNLL IIAKP* ORF13a and ORF13-1 show 94.4% identity in 126 aa overlap 10 20 30 40 50 60 orf13a. pep MTVWFVAAVAVLIIELLTGTVYLLWSAALAGSGIAYGLTGΞTPAAVLTAALLSALGIWF II II I I II II II I I II I I II II II II I I II II II II II I I I I I I II I I I orf13-1 AVLIIELLTGTVYLLWSAALAGSGIAYGLTGSTPAAVLTXALLSALGIXF 10 20 30 40 50 70 80 90 100 110 120 orf13a . pep VHAKTAVGKVETDSYQDLDAGQYAEILRHAGGNRYEVFYRGTHWQAQNTGQEELEPGTRA I II I I II II II I II I II II I I I : II II I : I I I I I II II II II II I I II II I I II II II I orf13-1 VHAKTAVRKVETDSYQDLDAGQYVEILRHTGGNRYEVFYRGTHWQAQNTGQEELEPGTRA 60 70 80 90 100 110 130 orf13a. pep LIVRKEGNLLIIAKPX I II II II I II IM: II orfl3-l LIVRKEGNLLIITHPX 120 Homology with a predicted ORF from Nsonorrhoeae ORF 13 shows 89.7% identity over a 126aa overlap with a predicted ORF (ORF13.ng) from N gonorrhoeae: orf l3 AVLIIELLTGTVYLLWSAALAGSGIAYGLTGSTPAAVLTXALLSALGIXF 51 I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I orfl3ng MTVWFVAAVAVLIIELLTGTVYLLWSAALAGSGIAYGLTGSTPAAVLTAALLSALGIWF 60 orfl3 VHAKTAVRKVETDSYQDLDAGQYVEILRHTGGΝRYEVXYRGTXWQAQΝTGQEELEPGTRA 111 I I I I I I I I I I I I I I I I I I : I : I : I I I M I I I I I I I I I I I I I I I I I I I I I M M M ! orfl3ng VHAKTAVGKVETDSYQDLDTGKYAEILRYTGGΝRYEVFYRGTHWQAQΝTGQEVFEPGTRA 120 orfl3 LIVRKEGΝLLIITHP 126 II I I I I I I II I M: I orf l3ng LLIIVVRRKKEEGGΝΝLLLLIIIIAAΝΝPP 135 The complete length ORF13ng nucleotide sequence <SEQ ID 65> is: 1 ATGACTGTAT GGTTTGTTGC CGCTGTTGCC GTCTTAATCA TCGAATTATT 51 GACGGGAACG GTTTATCTTT TGGTTGTCAG CGCGGCTTTG GCGGGTTCGG 101 GCATTGCCTA CGGGCTGACT GGCAGCACGC CTGCCGCCGT CTTGACCGCC 151 GCACTGCTTT CCGCGCTGGG CATTTGGTTC GTACATGCCA AAACCGCCGT 201 GGGAAAAGTT GAAACGGATT CATATCAGGA TTTGGATACC GGAAAATATG 251 CCGAAATCCT CCGATACACA GGCGGCAACC GTTACGAAGT TTTTTATCGC 301 GGTACGCACT GGCAGGCGCA AAATACGGGG CAGGAAGTGT TTGAACCGGG 351 AACGCGCGCC CTCATCGTCC GCAAAGAAGG TAACCTTCTT ATCATCGCAA 401 ACCCTTAA This encodes a protein having amino acid sequence <SEQ ID 66>: 1 MTVWFVAAVA VLIIELLTGT VYLLWSAAL AGSGIAYGLT GSTPAAVLTA 51 ALLSALGIWF VHAKTAVGKV ETDSYQDLDT GKYAEILRYT GGNRYEVFYR 101 GTHWQAQNTG QEVFEPGTRA LIVRKEGNLL IIANP* ORF13ng shows 91.3% identity in 126 aa overlap with ORF13-1 : 10 20 30 40 50 orf13-1. pep AVLIIELLTGTVYLLWSAALAGSGIAYGLTGSTPAAVLTXALLSALGIXF I I II II II I II I I I I I I I I I I II II II I II I I I II I I I II I I II I I I I I orfl3ng MTVWFVAAVAVLIIELLTGTVYLLWSAALAGSGIAYGLTGSTPAAVLTAALLSALGIWF 10 20 30 40 50 60 60 70 80 90 100 110 orf13-1. pep VHAKTAVRKVETDSYQDLDAGQYVEILRHTGGNRYEVFYRGTHWQAQNTGQEELEPGTRA I I II II I II II I I I I II I : I : I : I I I I : II II II I I I I I I II I II I I I II I : I I II I I orfl3ng VHAKTAVGKVETDSYQDLDTGKYAEILRYTGGNRYEVFYRGTHWQAQNTGQEVFEPGTRA 70 80 90 100 110 120 120 orf13-1. pep LIVRKEGNLLIITHPX II II I II I II I I :: II orfl3ng LIVRKEGNLLIIANPX 130 Based on this analysis, including the extensive leader sequence in this protein, it is predicted that ORF 13 and ORF 13ng are likely to be outer membrane proteins. It is thus predicted that the proteins from N. meningitidis .and N.gonorrhoeae, .and their epitopes, could be useful antigens for vaccines or diagnostics, or for raising antibodies. Example 9 The following DNA sequence was identified in N. meningitidis <SEQ ID 67>: 1 ATGTwTGATT TCGGTTTrGG CGArCTGGTT TTTGTCGGCA TTATCGCCCT 51 GATwGtCCTC GGCCCCGAAC GCsTGCCCGA GGCCGCCCGC AyCGCCGGAC 101 GGcTCATCGG CAGGCTGCAA CGCTTTGTCG GcAGCGTCAA ACAGGAATTT 151 GACACTCAAA TCGAACTGGA AGAACTGAGG AAGGCAAAGC AGGAATTTGA 201 AGCTGCCGcC GCTCAGGTTC GAGACAGCCT CAAAGAAACC GGTACGGATA 251 TGGAAGGCAA TCTGCACGAC ATTTCCGACG GTCTGAAGCC TTGGGAAAAA 301 CTGCCCGAAC AGCGGACACC TGCCGATTTC GGTGTCGATG AAAACGGCAA 351 TCCGCT.TCC CGATGCGGCA AACACCCTAT CAGACGGCAT TTCCGACGTT 401 ATGCCGTC. This corresponds to the amino acid sequence <SEQ ID 68; ORF2>: 1 MXDFGLGELV FVGIIALIVL GPERXPEAAR XAGRLIGRLQ RFVGSVKQEF 51 DTQIELEELR KAKQEFEAAA AQVRDSLKET GTDMEGNLHD ISDGLKPWEK 101 LPEQRTPADF GVDENGNPXS RCGKHPIRRH FRRYAV.. Further work revealed the complete nucleotide sequence <SEQ ID 69>: 1 ATGTTTGATT TCGGTTTGGG CGAGCTGGTT TTTGTCGGCA TTATCGCCCT 51 GATTGTCCTC GGCCCCGAAC GCCTGCCCGA GGCCGCCCGC ACCGCCGGAC 101 GGCTCATCGG CAGGCTGCAA CGCTTTGTCG GCAGCGTCAA ACAGGAATTT 151 GACACTCAAA TCGAACTGGA AGAACTGAGG AAGGCAAAGC AGGAATTTGA 201 AGCTGCCGCC GCTCAGGTTC GAGACAGCCT CAAAGAAACC GGTACGGATA 251 TGGAAGGCAA TCTGCACGAC ATTTCCGACG GTCTGAAGCC TTGGGAAAAA 301 CTGCCCGAAC AGCGGACACC TGCCGATTTC GGTGTCGATG AAAACGGCAA 351 TCCGCTTCCC GATGCGGCAA ACACCCTATC AGACGGCATT TCCGACGTTA 401 TGCCGTCCGA ACGTTCC AC GCTTCCGCCG AAACCCTTGG GGACAGCGGG 451 CAAACCGGCA GTACAGCCGA ACCCGCGGAA ACCGACCAAG ACCGCGCATG 501 GCGGGAATAC CTGACTGCTT CTGCCGCCGC ACCCGTCGTA CAGACCGTCG 551 AAGTCAGCTA TATCGATACT GCTGTTGAAA CGCCTGTTCC GCACACCACT 601 TCCCTGCGCA AACAGGCAAT AAGCCGCAAA CGCGATTTTC GTCCGAAACA 651 CCGCGCCAAA CCTAAATTGC GCGTCCGTAA ATCATAA This corresponds to the amino acid sequence <SEQ ID 70; ORF2-l>: 1 MFDFGLGELV FVGIIALIVL GPERLPEAAR TAGRLIGRLQ RFVGSVKQEF 51 DTQIELEELR KAKQEFEAAA AQVRDSLKET GTDMEGNLHD ISDGLKPWEK 101 LPEQRTPADF GVDENGNPLP DAANTLSDGI SDVMPSERSY ASAETLGDSG 151 QTGSTAEPAE TDQDRAWREY LTASAAAPW QTVEVSYIDT AVETPVPHTT 201 SLRKQAISRK RDFRPKHRAK PKLRVRKS* Further work identified the corresponding gene in strain A of N. meningitidis <SEQ LD 71 >: 1 ATGTTTGATT TCGGTTTGGG CGAGCTGGTT TTTGTCGGCA TTATCGCCCT 51 GATTGTCCTC GGCCCCGAAC GCCTGCCCGA GGCCGCCCGC ACCGCCGGAC 101 GGCTCATCGG CAGGCTGCAA CGCTTTGTCG GCAGCGTCAA ACAGGAATTT 151 GACACGCAAA TCGAACTGGA AGAACTAAGG AAGGCAAAGC AGGAATTTGA 201 AGCTGCCGCT GCTCAGGTTC GAGACAGCCT CAAAGAAACC GGTACGGATA 251 TGGAGGGTAA TCTGCACGAC ATTTCCGACG GTCTGAAGCC TTGGGAAAAA 301 CTGCCCGAAC AGCGCACGCC TGCTGATTTC GGTGTCGATG AAAACGGCAA 351 TCCCTTTCCC GATGCGGCAA ACACCCTATT AGACGGCATT TCCGACGTTA 401 TGCCGTCCGA ACGTTCCTAC GCTTCCGCCG AAACCCTTGG GGACAGCGGG 451 CAAACCGGCA GTACAGCCGA ACCCGCGGAA ACCGACCAAG ACCGTGCATG 501 GCGGGAATAC CTGACTGCTT CTGCCGCCGC ACCCGTCGTA CAGACCGTCG 551 AAGTCAGCTA TATCGATACC GCTGTTGAAA CCCCTGTTCC GCATACCACT 601 TCGCTGCGTA AACAGGCAAT AAGCCGCAAA CGCGATTTGC GTCCTAAATC 651 CCGCGCCAAA CCTAAATTGC GCGTCCGTAA ATCATAA This encodes a protein having amino acid sequence <SEQ ID 72; ORF2a>: 1 MFDFGLGELV FVGIIALIVL GPERLPEAAR TAGRLIGRLQ RFVGSVKQEF 51 DTQIELEELR KAKQEFEAAA AQVRDSLKET GTDMEGNLHD ISDGLKPWEK 101 LPEQRTPADF GVDENGNPFP DAANTLLDGI SDVMPSERSY ASAETLGDSG 151 QTGSTAEPAE TDQDRAWREY LTASAAAPW QTVEVSYIDT AVETPVPHTT 201 SLRKQAISRK RDLRPKSRAK PKLRVRKS* The originally-identified partial strain B sequence (ORF2) shows 97.5% identity over a 118aa overlap with ORF2a: 10 20 30 40 50 60 orf2. pep MXDFGLGELVFVGIIALIVLGPERXPEAARXAGRLIGRLQRFVGSVKQEFDTQIELEELR I II II I I II II II I II I I I I II I I II I I : II I II I I I I I I I II I I II I II I II I II II orf2a MFDFGLGELVFVGIIALIVLGPERLPEAARTAGRLIGRLQRFVGSVKQEFDTQIELEELR 10 20 30 40 50 60 70 80 90 100 110 120 orf2. pep KAKQEFEAAAAQVRDSLKETGTDMEGNLHDISDGLKPWEKLPEQRTPADFGVDENGNPXS I I I II II II I I I I II I I I I II I I II I I I I I II II I I I I I II II I I II I I I I I I I I I I I orf2a KAKQEFEAAAAQVRDSLKETGTDMEGNLHDISDGLKPWEKLPEQRTPADFGVDENGNPFP 70 80 90 100 110 120 130 orf2. pep RCGKHPIRRHFRRYAV orf2a DAANTLLDGISDVMPSERSYASAETLGDSGQTGSTAEPAETDQDRAWREYLTASAAAPW 130 140 150 160 170 180 The complete strain B sequence (ORF2-1) and ORF2a show 98.2% identity in 228 aa overlap: orf2a. pep MFDFGLGELVFVGIIALIVLGPERLPEAARTAGRLIGRLQRFVGSVKQEFDTQIELEELR 60 I I I II I II I II I I I I II I I II I I I II II I II I I II I I I I I I I I I II II I I I I I I I I I II I orf2-l MFDFGLGELVFVGIIALIVLGPERLPEAARTAGRLIGRLQRFVGSVKQEFDTQIELEELR 60 orf2a. pep KAKQEFEAAAAQVRDSLKETGTDMEGNLHDISDGLKPWEKLPEQRTPADFGVDENGNPFP 120 I II I I I I I II II I II I II I I I II I I I I I II II I I II I I I II I II I I I I I II I II I II I: I orf2-l KAKQEFEAAAAQVRDSLKETGTDMEGNLHDISDGLKPWEKLPEQRTPADFGVDENGNPLP 120 orf2a. pep DAANTLLDGISDVMPSERSYASAETLGDSGQTGSTAEPAETDQDRAWREYLTASAAAPW If II II II I I II I II I II I II II II I I I I I II I II I I I I I I I I I I I I I II I I II I I I I M I orf2-l DAANTLSDGISDVMPSERSYASAETLGDSGQTGSTAEPAETDQDRAWREYLTASAAAPW 180 orf2a. pep QTVEVSYIDTAVETPVPHTTSLRKQAISRKRDLRPKSRAKPKLRVRKSX 229 II I I II I I I II II I II I I II I II II II I I II I : II I I I I I II II II II orf2-1 QTVEVSYIDTAVETPVPHTTSLRKQAISRKRDFRPKHRAKPKLRVRKSX 229 Further work identified a partial DNA sequence <SEQ ID 73> in N.gonorrhoeae encoding the following amino acid sequence <SEQ ID 74; ORF2ng>: 1 MFDFGLGELI FVGIIALIVL GPERLPEAAR TAGRLIGRLQ RFVGSVKQEL 51 DTQIELEELR KVKQAFEAAA AQVRDSLKET DTDMQNSLHD ISDGLKPWEK 101 LPEQRTPADF GVDEKGNSLS RYGKHRIRRH FRRYAV* Further work identified the complete gonococcal gene sequence <SEQ ID 75>: 1 ATGTTTGATT TCGGTTTGGG CGAGCTGATT TTTGTCGGCA TTATCGCCCT 51 GATTGTCCTT GGTCCAGAAC GCCTGCCCGA AGCCGCCCGC ACTGCCGGAC 101 GGCTTATCGG CAGGCTGCAA CGCTTTGTAG GAAGCGTCAA ACAAGAACTT 151 GACACTCAAA TCGAACTGGA AGAGCTGAGG AAGGTCAAGC AGGCATTCGA 201 AGCTGCCGCC GCTCAGGTTC GAGACAGCCT CAAAGAAACC GATACGGATA 251 TGCAGAACAG TCTGCACGAC ATTTCCGACG GTCTGAAGCC TTGGGAAAAA 301 CTGCCCGAAC AGCGCACGCc tgccgatttc gGTGTCGATg AAAacggcaa 351 tccccttccc gATACGGCAA ACACCGTATC AGACGGCATT TCCGACGTTA 401 TGCCGTCTGA ACGTTCCGAT ACTtccgcCG AAACCCTTGG GGACGACAGG 451 CAAACCGGCA GTACAGCCGA ACCTGCGGAA ACCGACAAAG ACCGCGCATG 501 GCGGGAATAC CTGactgctt ctgccgccgc acctgtcgta Cagagggccg 551 tcgaagtcag ctaTATCGAT ACTGCTGTTG AAacgcctgT tccgcaCacc 601 acttccctgc gcaAACAGGC AATAAACCGC AAACGCGATT TttgtccgaA 651 ACACCGCGCc aAACCGAAat tgcgcgtcCG TAAATCATAA This encodes a protein having the amino acid sequence <SEQ ID 76; ORF2ng-l>: 1 MFDFGLGELI FVGIIALIVL GPERLPEAAR TAGRLIGRLQ RFVGSVKQEL 51 DTQIELEELR KVKQAFEAAA AQVRDSLKET DTDMQNSLHD ISDGLKPWEK 101 LPEQRTPADF GVDENGNPLP DTANTVSDGI SDVMPSERSD TSAETLGDDR 151 QTGSTAEPAE TDKDRAWREY LTASAAAPW QRAVEVSYID TAVETPVPHT 201 TSLRKQAINR KRDFCPKHRA KPKLRVRKS* The originally-identified partial strain B sequence (ORF2) shows 87.5% identity over a 136aa overlap with ORF2ng: orf2. pep MXDFGLGELVFVGIIALIVLGPERXPEAARXAGRLIGRLQRFVGSVKQEFDTQIELEELR 60 I I I II I IM I I I I I I I I I II I II I I II I : I I II II II II I I I II II I: II II II I I I I orf2ng MFDFGLGELIFVGIIALIVLGPERLPEAARTAGRLIGRLQRFVGSVKQELDTQIELEELR 60 orf2.pep KAKQEFEAAAAQVRDSLKETGTDMEGNLHDISDGLKPWEKLPEQRTPADFGVDENGNPXS 120 MM II I II I II II II II I II I : : : I II I I II II II II II I II II I II I II I : I I orf2ng KVKQAFEAAAAQVRDSLKETDTDMQNSLHDISDGLKPWEKLPEQRTPADFGVDEKGNSLP 120 orf2. pep RCGKHPIRRHFRRYAV 136 I III II I II II I II orf2ng RYGKHRIRRHFRRYAV 136 The complete strain B and gonococcal sequences (ORF2-1 & ORF2ng-l) show 91.7% identity in 229 aa overlap: 10 20 30 40 50 60 orf2-1. pep MFDFGLGELVFVGIIALIVLGPERLPEAARTAGRLIGRLQRFVGSVKQEFDTQIELEELR I M II II II : I I II I I I II I I II I I I II I I II I I I I II II I I I I I I II I : I I I I I II II I orf2ng-l MFDFGLGELIFVGIIALIVLGPERLPEAARTAGRLIGRLQRFVGSVKQELDTQIELEELR 10 20 30 40 50 60 70 80 90 100 110 120 orf2-1.pep KAKQEFEAAAAQVRDSLKETGTDMEGNLHDISDGLKPWEKLPEQRTPADFGVDENGNPLP MM I II I II I I I I I I II I II I ::: I | | I I I I I I II I II I I I I II I II II I I I II II I orf2ng-l KVKQAFEAAAA Q VRDSLKETDTDMQNSLHDISDGLKPWEKLPEQRTPADFGVDENGNPLP 70 80 90 100 110 120 130 140 150 160 170 180 orf2-1. pep DAANTLSDGISDVMPSERSYASAETLGDSGQTGSTAEPAETDQDRAWREYLTASAAAPW I : I I I : I II I II II II I I I : I II I II I : I II I I II I II I I : I I I I I I II I I II I I II I orf2ng-l DTANTVSDGISDVMPSERSDTSAETLGDDRQTGSTAEPAETDKDRAWREYLTASAAAPW 130 140 150 160 170 180 190 200 210 220 229 orf2-1. pep Q-TVEVSYIDTAVETPVPHTTSLRKQAISRKRDFRPKHRAKPKLRVRKSX I : I II I II I I I I I II I I I I I I I II I I I : II I II I II I I I I II I II I I I orf2ng-l QRAVEVSYIDTAVETPVPHTTSLRKQAINRKRDFCPKHRAKPKLRVRKSX 190 200 210 220 230 Computer analysis of these amino acid sequences indicates a transmembrane region (underlined), .and also revealed homology (59% identity) between the gonococcal sequence and the TatB protein of E.coli: gnl|PID|el292181 (AJ005830) TatB protein [Escherichia coli] Length = 171 Score = 56.6 bits (134), Expect = le-07 Identities = 30/88 (34%), Positives = 52/88 (59%), Gaps = 1/88 (1%) Query: 1 MFDFGLGELIFVGIIALIVLGPERLPEAARTAGRLIGRLQRFVGSVKQELDTQIELEELR 60 MFD G EL+ V II L+VLGP+RLP A +T I L+ +V+ EL +++L+E + Sbjct: 1 MFDIGFSELLLVFIIGLWLGPQRLPVAVKTVAGWIRALRSLATTVQNELTQELKLQEFQ 60 Query: 61 -KVKQAFEAAAAQVRDSLKETDTDMQNS 87 +K+ +A+ + LK + +++ + Sbjct: 61 DSLKKVEKASLTNLTPELKASMDELRQA 88 Based on this analysis, it was predicted that ORF2, ORF2a and ORF2ng are likely to be membrane proteins and so the proteins from N. meningitidis and N.gonorrhoeae, and their epitopes, could be useful antigens for vaccines or diagnostics, or for raising antibodies. ORF2-1 (16kDa) was cloned in pET and pGex vectors and expressed in E.coli, as described above. The products of protein expression and purification were analyzed by SDS-PAGE. Figure 3A shows the results of affinity purification of the GST-fusion protein, .and Figure 3B shows the results of expression of the His-fusion in E.coli. Purified GST-fusion protein was used to immunise mice, whose sera were used for Western blots (Figure 3C), ELISA (positive result), and FACS analysis (Figure 3D). These experiments confirm that ORF37-1 is a surface-exposed protein, and that it is a useful immunogen. Example 10 The following partial DNA sequence was identified in N. meningitidis <SEQ ID 77>: 1 ATGCAAGCAC GGCTGCTGAT ACCTATTCTT TTTTCAGTTT TTATTTTATC 51 CGC.TGCGGG ACACTGACAG GTATTCCATC GCATGGCGgA GkTAAACgCT 101 TTgCGGTCGA ACAAGAACTT GTGGCCGCTT CTGCCAGAGC TGCCGTTAAA 151 GACATGGATT TACAGGCATT ACACGGACGA AAAGTTGCAT TGTACATTGC 201 CACTATGGGC GACCAAGGTT CAGGcAGTTT GACAGGGGGG TCGCTACTCC 251 ATTGATGCAC kGrT CsTGG CGAATACATA AACAGCCCTG CCGTCCGTAC 301 CGATTACACC TATCCACGTT ACGAAACCAC CGCTGAAACA ACATCAGGCG 351 GTTTGACAGG TTTAACCACT TCTTTATCTA CACTTAATGC CCCTGCACTC 401 TCTCGCACCC AATCAGACGG TAGCGGAAGT AAAAGCAGTC TGGGCTTAAA 451 TATTGGCGGG ATGGGGGATT ATCGAAATGA AACCTTGACG ACTAACCCGC 501 GCGACACTGC CTTTCTTTCC CACTTGGTAC AGACCGTATT TTTCCTGCGC 551 GGCATAGACG TTGTTTCTCC TGCCAATGCC GATACAGATG TGTTTATTAA 601 CATCGACGTA TTCGGAACGA TACGCAACAG AACCGAAATG.. This corresponds to the amino acid sequence <SEQ ID 78; ORF15>: 1 MQARLLIPIL FSVFILSACG TLTGIPSHGG XKRFAVEQEL VAASARAAVK 51 DMDLQALHGR KVALYIATMG DQGSGSLTGG RYSIDAXXXG EYINSPAVRT 101 DYTYPRYETT AETTSGGLTG LTTSLSTLNA PALSRTQSDG SGSKSSLGLN 151 IGGMGDYRNE TLTTNPRDTA FLSHLVQTVF FLRGIDWSP ANADTDVFIN 201 IDVFGTIRNR TEM.. Further work revealed the complete nucleotide sequence <SEQ ID 79>: 1 ATGCAAGCAC GGCTGCTGAT ACCTATTCTT TTTTCAGTTT TTATTTTATC 51 CGCCTGCGGG ACACTGACAG GTATTCCATC GCATGGCGGA GGTAAACGCT 101 TTGCGGTCGA ACAAGAACTT GTGGCCGCTT CTGCCAGAGC TGCCGTTAAA 151 GACATGGATT TACAGGCATT ACACGGACGA AAAGTTGCAT TGTACATTGC 201 CACTATGGGC GACCAAGGTT CAGGCAGTTT GACAGGGGGT CGCTACTCCA 251 TTGATGCACT GATTCGTGGC GAATACATAA ACAGCCCTGC CGTCCGTACC 301 GATTACACCT ATCCACGTTA CGAAACCACC GCTGAAACAA CATCAGGCGG 351 TTTGACAGGT TTAACCACTT CTTTATCTAC ACTTAATGCC CCTGCACTCT 401 CTCGCACCCA ATCAGACGGT AGCGGAAGTA AAAGCAGTCT GGGCTTAAAT 451 ATTGGCGGGA TGGGGGATTA TCGAAATGAA ACCTTGACGA CTAACCCGCG 501 CGACACTGCC TTTCTTTCCC ACTTGGTACA GACCGTATTT TTCCTGCGCG 551 GCATAGACGT TGTTTCTCCT GCCAATGCCG ATACAGATGT GTTTATTAAC 601 ATCGACGTAT TCGGAACGAT ACGCAACAGA ACCGAAATGC ACCTATACAA 651 TGCCGAAACA CTGAAAGCCC AAACAAAACT GGAATATTTC GCAGTAGACA 701 GAACCAATAA AAAATTGCTC ATCAAACCAA AAACCAATGC GTTTGAAGCT 751 GCCTATAAAG AAAATTACGC ATTGTGGATG GGGCCGTATA AAGTAAGCAA 801 AGGAATTAAA CCGACGGAAG GATTAATGGT CGATTTCTCC GATATCCGAC 851 CATACGGCAA TCATACGGGT AACTCCGCCC CATCCGTAGA GGCTGATAAC 901 AGTCATGAGG GGTATGGATA CAGCGATGAA GTAGTGCGAC AACATAGACA 951 AGGACAACCT TGA This corresponds to the amino acid sequence <SEQ ID 80; ORF15-l>: 1 MQARLLIPIL FSVFILSACG TLTGIPSHGG GKRFAVEQEL VAASARAAVK 51 DMDLQALHGR KVALYIATMG DQGSGSLTGG RYSIDALIRG EYINSPAVRT 101 DYTYPRYETT AETTSGGLTG LTTSLSTLNA PALSRTQSDG SGSKSSLGLN 151 IGGMGDYRNE TLTTNPRDTA FLSHLVQTVF FLRGIDWSP ANADTDVFIN 201 IDVFGTIRNR TEMHLYNAET LKAQTKLEYF AVDRTNKKLL IKPKTNAFEA 251 AYKENYALWM GPYKVSKGIK PTEGLMVDFS DIRPYGNHTG NSAPSVEADN 301 SHEGYGYSDE WRQHRQGQP * Further work identified the coπesponding gene in strain A of N. meningitidis <SEQ LD 81>: 1 ATGCAAGCAC GGCTGCTGAT ACCTATTCTT TTTTCAGTTT TTATTTTATC 51 CGCCTGCGGG ACACTGACAG GTATTCCATC GCATGGCGGA GGTAAACGCT 101 TTGCGGTCGA ACAAGAACTT GTGGCCGCTT CTGCCAGAGC TGCCGTTAAA 151 GACATGGATT TACAGGCATT ACACGGACGA AAAGTTGCAT TGTACATTGC 201 AACTATGGGC GACCAAGGTT CAGGCAGTTT GACAGGGGGT CGCTACTCCA 251 TTGATGCACT GATTCGTGGC GAATACATAA ACAGCCCTGC CGTCCGTACC 301 GATTACACCT ATCCACGTTA CGAAACCACC GCTGAAACAA CATCAGGCGG 351 TTTGACAGGT TTAACCACTT CTTTATCTAC ACTTAATGCC CCTGCACTCT 401 CGCGCACCCA ATCAGACGGT AGCGGAAGTA AAAGCAGTCT GGGCTTAAAT 451 ATTGGCGGGA TGGGGGATTA TCGAAATGAA ACCTTGACGA CTAACCCGCG 501 CGACACTGCC TTTCTTTCCC ACTTGGTACA GACCGTATTT TTCCTGCGCG 551 GCATAGACGT TGTTTCTCCT GCCAATGCCG ATACGGATGT GTTTATTAAC 601 ATCGACGTAT TCGGAACGAT ACGCAACAGA ACCGAAATGC ACCTATACAA 651 TGCCGAAACA CTGAAAGCCC AAACAAAACT GGAATATTTC GCAGTAGACA 701 GAACCAATAA AAAATTGCTC ATCAAACCAA AAACCAATGC GTTTGAAGCT 751 GCCTATAAAG AAAATTACGC ATTGTGGATG GGACCGTATA AAGTAAGCAA 801 AGGAATTAAA CCGACAGAAG GATTAATGGT CGATTTCTCC GATATCCAAC 851 CATACGGCAA TCATATGGGT AACTCTGCCC CATCCGTAGA GGCTGATAAC 901 AGTCATGAGG GGTATGGATA CAGCGATGAA GCAGTGCGAC GACATAGACA 951 AGGGCAACCT TGA This encodes a protein having amino acid sequence <SEQ LD 82; ORF15a>: 1 MQARLLIPIL FSVFILSACG TLTGIPSHGG GKRFAVEQEL VAASARAAVK 51 DMDLQALHGR KVALYIATMG DQGSGSLTGG RYSIDALIRG EYINSPAVRT 101 DYTYPRYETT AETTSGGLTG LTTSLSTLNA PALSRTQSDG SGSKSSLGLN 151 IGGMGDYRNE TLTTNPRDTA FLSHLVQTVF FLRGIDWSP ANADTDVFIN 201 IDVFGTIRNR TEMHLYNAET LKAQTKLEYF AVDRTNKKLL IKPKTNAFEA 251 AYKENYALWM GPYKVSKGIK PTEGLMVDFS DIQPYGNHMG NSAPSVEADN 301 SHEGYGYSDE AVRRHRQGQP * The originally-identified partial strain B sequence (ORF15) shows 98.1% identity over a 213aa overlap with ORF 15 a: 10 20 30 40 50 60 orf15. pep MQARLLIPILFSVFILSACGTLTGIPSHGGXKRFAVEQELVAASARAAVKDMDLQALHGR I II I II I II I II I I II II II I I II I I II I I II I I II I I I I II I II II I I I II II I I I I I orf15a MQARLLIPILFSVFILSACGTLTGIPSHGGGKRFAVEQELVAASARAAVKDMDLQALHGR 10 20 30 40 50 60 70 80 90 100 110 120 orf15. pep KVALYIATMGDQGSGSLTGGRYSIDAXXXGEYINSPAVRTDYTYPRYETTAETTSGGLTG I II I II I I II II II II II I II I I II I I II I I II II I I I I II I II I II II II I I I I I I orf15a KVALYIATMGDQGSGSLTGGRYSIDALIRGEYINSPAVRTDYTYPRYETTAETTSGGLTG 70 80 90 100 110 120 130 140 150 160 170 180 orf15. pep LTTSLSTLNAPALSRTQSDGSGSKSSLGLNIGGMGDYRNETLTTNPRDTAFLSHLVQTVF I II I II I I II II I I I II II I II I I II I II II I II I I I I II II I I II II I I II I I II I I I I orf15a LTTSLSTLNAPALSRTQSDGSGSKSSLGLNIGGMGDYRNETLTTNPRDTAFLSHLVQTVF 130 140 150 160 170 180 190 200 210 orf15. pep FLRGIDWSPANADTDVFINIDVFGTIRNRTEM I I II I I I I I II I I I II II II I I I I I I I I I I I I I orf15a FLRGIDWSPANADTDVFINIDVFGTIRNRTEMHLYNAETLKAQTKLEYFAVDRTNKKLL 190 200 210 220 230 240 The complete strain B sequence (ORF15-1) and ORF15a show 98.8% identity in 320 aa overlap: 10 20 30 40 50 60 orf15a . pep MQARLLIPILFSVFILSACGTLTGIPSHGGGKRFAVEQELVAASARAAVKDMDLQALHGR I I II I I II II I I I II I I I I II I I II I II II I II I I I I I I I II I I I I I I I I I II I I I I I II orf15-1 MQARLLIPILFSVFILSACGTLTGIPSHGGGKRFAVEQELVAASARAAVKDMDLQALHGR 10 20 30 40 50 60 70 80 90 100 110 120 orf15a. pep KVALYIATMGDQGSGSLTGGRYSIDALIRGEYINSPAVRTDYTYPRYETTAETTSGGLTG II I I I I I II I II I I I II I II I I I I I I I II II II I I II I I I I I I I I I I II I II I I I I I II I orf15-1 KVALYIATMGDQGSGSLTGGRYSIDALIRGEYINSPAVRTDYTYPRYETTAETTSGGLTG 70 80 90 100 110 120 130 140 150 160 170 180 orf15a. pep LTTSLSTLNAPALSRTQSDGSGSKSSLGLNIGGMGDYRNETLTTNPRDTAFLSHLVQTVF I I I II I I I I II II I II I II I I I II I II II II I I II II I II II I II I I I I I I I I I I I I I I I orf15-1 LTTSLSTLNAPALSRTQSDGSGSKSSLGLNIGGMGDYRNETLTTNPRDTAFLSHLVQTVF 130 140 150 160 170 180 190 200 210 220 230 240 orf15 . pep FLRGIDWSPANADTDVFINIDVFGTIRNRTEMHLYNAETLKAQTKLEYFAVDRTNKKLL I II I II I I II II I I II I I I I II I I I I I II I II II II I I II I I II II II I I II II II I I I I orf15-1 FLRGIDWSPANADTDVFINIDVFGTIRNRTEMHLYNAETLKAQTKLEYFAVDRTNKKLL 190 200 210 220 230 240 250 260 270 280 290 300 orf15a . pep IKPKTNAFEAAYKENYALWMGPYKVSKGIKPTEGLMVDFSDIQPYGNHMGNSAPSVEADN I I II I I II I II I I I I II I II II II II II II I I I I I I II I II M I I II I II I II I I I I II orf15-1 IKPKTNAFEAAYKENYALWMGPYKVSKGIKPTEGLMVDFSDIRPYGNHTGNSAPSVEADN 250 260 270 280 290 300 310 320 orf15a .pep SHEGYGYSDEAVRRHRQGQPX II I I I II II I : II : II I II I I orfl5-l SHEGYGYSDEWRQHRQGQPX 310 320 Further work identified the corresponding gene in N.gonorrhoeae <SEQ ID 83>: 1 ATGCGGGCAC GGCTGCTGAT ACCTATTCTT TTTTCAGTTT TTATTTTATC 51 CGCCTGCGGG ACACTGACAG GTATTCCATC GCATGGCGGA GGCAAACGCT 101 TCGCGGTCGA ACAAGAACTT GTGGCCGCTT CTGCCAGAGC TGCCGTTAAA 151 GACATGGATT TACAGGCATT ACACGGACGA AAAGTTGCAT TGTACATTGC 201 AACTATGGGC GACCAAGGTT CAGGCAGTTT GACAGGGGGT CGCTACTCCA 251 TTGATGCACT GATTCGCGGC GAATACATAA ACAGCCCTGC CGTCCGCACC 301 GATTACACCT ATCCGCGTTA CGAAACCACC GCTGAAACAA CATCAGGCGG 351 TTTGACGGGT TTAACCACTT CTTTATCTAC ACTTAATGCC CCTGCACTCT 401 CGCGCACCCA ATCAGACGGT AGCGGAAGTA GGAGCAGTCT GGGCTTAAAT 451 ATTGGCGGGA TGGGGGATTA TCGAAATGAA ACCTTGACGA CCAACCCGCG 501 CGACACTGCC TTTCTTTCCC ACTTGGTGCA GACCGTATTT TTCCTGCGCG 551 GCATAGACGT TGTTTCTCCT GCCAATGCCG ATACAGATGT GTTTATTAAC 601 ATCGACGTAT TCGGAACGAT ACGCAACAGA ACCGAAATGC ACCTATACAA 651 TGCCGAAACA CTGAAAGCCC AAACAAAACT GGAATATTTC GCAGTAGACA 701 GAACCAATAA AAAATTGCTC ATCAAACCCA AAACCAATGC GTTTGAAGCT 751 GCCTATAAAG AAAATTACGC ATTGTGGATG GGGCCGTATA AAGTAAGCAA 801 AGGAATCAAA CCGACGGAAG GATTGATGGT CGATTTCTCC GATATCCAAC 851 CATACGGCAA TCATACGGGT AACTCCGCCC CATCCGTAGA GGCTGATAAC 901 AGTCATGAGG GGTATGGATA CAGCGATGAA GCAGTGCGAC AACATAGACA 951 AGGGCAACCT TGA This encodes a protein having amino acid sequence <SEQ ID 84; ORF15ng>: 1 MRARLLIPIL FSVFILSACG TLTGIPSHGG GKRFAVEQEL VAASARAAVK 51 DMDLQALHGR KVALYIATMG DQGSGSLTGG RYSIDALIRG EYINSPAVRT 101 DYTYPRYETT AETTSGGLTG LTTSLSTLNA PALSRTQSDG SGSRSSLGLN 151 IGGMGDYRNE TLTTNPRDTA FLSHLVQTVF FLRGIDWSP ANADTDVFIN 201 IDVFGTIRNR TEMHLYNAET LKAQTKLEYF AVDRTNKKLL IKPKTNAFEA 251 AYKENYALWM GPYKVSKGIK PTEGLMVDFS DIQPYGNHTG NSAPSVEADN 301 SHEGYGYSDE AVRQHRQGQP * The originally-identified partial strain B sequence (ORF15) shows 97.2% identity over a 213aa overlap with ORF15ng: orf15. pep MQARLLIPILFSVFILSACGTLTGIPSHGGXKRFAVEQELVAASARAAVKDMDLQALHGR 60 I : I II II II I II I I II I I I I II I I I I I I II II I I I II I II II II I I II I I II II I II I I orfl5ng MRARLLIPILFSVFILSACGTLTGIPSHGGGKRFAVEQELVAASARAAVKDMDLQALHGR 60 orf15.pep KVALYIATMGDQGSGSLTGGRYSIDAXXXGEYINSPAVRTDYTYPRYETTAETTSGGLTG 120 I II I II I II I I I I I II II I II I I II I I II I II II I I I II II II II I I I I I I II I I I I orf15ng KVALYIATMGDQGSGSLTGGRYSIDALIRGEYINSPAVRTDYTYPRYETTAETTSGGLTG 120 orfl5.pep LTTSLSTLNAPALSRTQSDGSGSKSSLGLNIGGMGDYRNETLTTNPRDTAFLSHLVQTVF 180 II I II I I I II II II I II I I I I I I : I I II I I I I II II II I I II I II II II I II II II I I I I orflδng LTTSLSTLNAPALSRTQSDGSGSRSSLGLNIGGMGDYRNETLTTNPRDTAFLSHLVQTVF 180 orf15. pep FLRGIDWSPANADTDVFINIDVFGTIRNRTEM 213 II II II II I II II II I I I I I II I I I II I I II I I orfl5ng FLRGIDWSPANADTDVFINIDVFGTIRNRTEMHLYNAETLKAQTKLEYFAVDRTNKKLL 240 The complete strain B sequence (ORF15-1) and ORF15ng show 98.8% identity in 320 aa overlap: 10 20 30 40 50 60 orf15-1. pep MQARLLIPILFSVFILSACGTLTGIPSHGGGKRFAVEQELVAASARAAVKDMDLQALHGR I : II I I I I I II I II I I I I II I I I II I I II I II I I II I I II I I I II II I I I II I II I I II I orfl5ng MRARLLIPILFSVFILSACGTLTGIPSHGGGKRFAVEQELVAASARAAVKDMDLQALHGR 10 20 30 40 50 60 70 80 90 100 110 120 orf15-1. pep KVALYIATMGDQGSGSLTGGRYSIDALIRGEYINSPAVRTDYTYPRYETTAETTSGGLTG I II I I II II II II II I I I II II I I II I I I I II I II II I II II I I II I II I II I I II I II I orfl5ng KVALYIATMGDQGSGSLTGGRYSIDALIRGEYINSPAVRTDYTYPRYETTAETTSGGLTG 70 80 90 100 110 120 130 140 150 160 170 180 orf15-1. pep LTTSLSTLNAPALSRTQSDGΞGSKSSLGLNIGGMGDYRNETLTTNPRDTAFLSHLVQTVF I I I I I I I I II I II II II I I I I I I : I I I I II II II I II I I I I I I I II I I I II I I I I I I I II orf15ng LTTSLSTLNAPALSRTQSDGSGSRSSLGLNIGGMGDYRNETLTTNPRDTAFLSHLVQTVF 130 140 150 160 170 180 190 200 210 220 230 240 orf15-1. pep FLRGIDWSPANADTDVFINIDVFGTIRNRTEMHLYNAETLKAQTKLEYFAVDRTNKKLL I II II II I II II I II II II II I II I II I I I II I I I I I I I II I II I I II II II II I II II I orfl5ng FLRGIDVVSPANADTDVFINIDVFGTIRNRTEMHLYNAETLKAQTKLEYFAVDRTNKKLL 190 200 210 220 230 240 250 260 270 280 290 300 orf15-1. pep IKPKTNAFEAAYKENYALWMGPYKVSKGIKPTEGLMVDFSDIRPYGNHTGNSAPSVEADN I I II I I I I I I I II I II II I I I II I II II II I I II I II I I II I : I II II II II I I I I I II I orfl5ng IKPKTNAFEAAYKENYALWMGPYKVSKGIKPTEGLMVDFSDIQPYGNHTGNSAPSVEADN 250 260 270 280 290 300 310 320 orf15-1. pep SHEGYGYSDEWRQHRQGQPX I I I I I I I I I I : I I I I I I I I I I orfl5ng SHEGYGYSDEAVRQHRQGQPX 310 320 Computer analysis of these amino acid sequences reveals an ILSAC motif (putative membrane lipoprotein lipid attachment site, as predicted by the MOTIFS program). indicates a putative leader sequence, and it was predicted that the proteins from N. meningitidis and N.gonorrhoeae, and their epitopes, could be useful antigens for vaccines or diagnostics, or for raising antibodies. ORF 15-1 (31.7kDa) was cloned in pET and pGex vectors and expressed in E.coli, as described above. The products of protein expression and purification were analyzed by SDS-PAGE. Figure 4A shows the results of affinity purification of the GST-fusion protein, and Figure 4B shows the results of expression of the His-fusion in E.coli. Purified GST-fusion protein was used to immunise mice, whose sera were used for Western blot (Figure 4C) and ELISA (positive result). These experiments confirm that ORFX-1 is a surface-exposed protein, and that it is a useful immunogen. Example 11 The following partial DNA sequence was identified in N. meningitidis <SEQ ID 85>: 1 ..GG.CAGCACA AAAAACAGGC GGTTGAACGG AAAAACCGTA TTTACGATGA 51 TGCCGGGTAT GATATTCGGC GTATTCACGG GCGCATTCTC CGCAAAATAT 101 ATCCCCGCGT TCGGGCTTCA AATTTTCTTC ATCCTGTTTT TAACCGCCGT 151 CGCATTCAAA ACACTGCATA CCGACCCTCA GACGGCATCC CGCCCGCTGC 201 CCGGACTGCC CrGACTGACT GCGGTTTCCA CACTGTTCGG CACAATGTCG 251 AGCTGGGTCG GCATAGGCGG CGGTTCACTT TCCGTCCCCT TCTTAATCCA 301 CTGCGGCTTC CCCGCCCATA AAGCCATCGG CACATCATCC GGCCTTGCCT 351 GGCCGATTGC ACTCTCCGGC GCAATATCGT ATCTGCTCAA CGGCCTGAAT 401 ATTGCAGGAT TGCCCGAAGG GTCACTGGGC TTCCTTTACC TGCCCGCCGT 451 CGCCGTCCTC AGCGCGGCAA CCATTGCCTT TGCCCCGCTC GGTGTCAAAA 501 CCGCCCACAA ACTTTCTTCT GCCAAACTCA AAAAATC.TT CGGCATTATG 551 TTGCTTTTGA TTGCCGGAAA AATGCTGTAC AACCTGCTTT AA This corresponds to the amino acid sequence <SEQ ID 86; ORF17>: 1 ..GQHKKQAVNG KTVFTMMPGM IFGVFTGAFS AKYIPAFGLQ IFFILFLTAV 51 AFKTLHTDPQ TASRPLPGLP XLTAVSTLFG TMSSWVGIGG GSLSVPFLIH 101 CGFPAHKAIG TSSGLAWPIA LSGAISYLLN GLNIAGLPEG SLGFLYLPAV 151 AVLSAATIAF APLGVKTAHK LSSAKLKKSF GIMLLLIAGK MLYNLL* Further work revealed the complete nucleotide sequence <SEQ ID 87>: 1 ATGTGGCATT GGGACATTAT CTTAATCCTG CTTGCCGTAG GCAGTGCGGC 51 AGGTTTTATT GCCGGCCTGT TCGGCGTAGG CGGCGGCACG CTGATTGTCC 101 CTGTCGTTTT ATGGGTGCTT GATTTGCAGG GTTTGGCACA ACATCCTTAC 151 GCGCAACACC TCGCCGTCGG CACATCCTTC GCCGTCATGG TCTTCACCGC 201 CTTTTCCAGT ATGCTGGGGC AGCACAAAAA ACAGGCGGTC GACTGGAAAA 251 CCGTATTTAC GATGATGCCG GGTATGATAT TCGGCGTATT CACGGGCGCA 301 CTCTCCGCAA AATATATCCC CGCGTTCGGG CTTCAAATTT TCTTCATCCT 351 GTTTTTAACC GCCGTCGCAT TCAAAACACT GCATACCGAC CCTCAGACGG 401 CATCCCGCCC GCTGCCCGGA CTGCCCGGAC TGACTGCGGT TTCCACACTG 451 TTCGGCACAA TGTCGAGCTG GGTCGGCATA GGCGGCGGTT CACTTTCCGT 501 CCCCTTCTTA ATCCACTGCG GCTTCCCCGC CCATAAAGCC ATCGGCACAT 551 CATCCGGCCT TGCCTGGCCG ATTGCACTCT CCGGCGCAAT ATCGTATCTG 601 CTCAACGGCC TGAATATTGC AGGATTGCCC GAAGGGTCAC TGGGCTTCCT 651 TTACCTGCCC GCCGTCGCCG TCCTCAGCGC GGCAACCATT GCCTTTGCCC 701 CGCTCGGTGT CAAAACCGCC CACAAACTTT CTTCTGCCAA ACTCAAAAAA 751 Tc.TTCGGCA TTATGTTGCT TTTGATTGCC GGAAAAATGC TGTACAACCT 801 GCTTTAA This corresponds to the amino acid sequence <SEQ ID 88; ORF17-l>: 1 MWHWDIILIL LAVGSAAGFI AGLFGVGGGT LIVPWLWVL DLQGLAQHPY 51 AQHLAVGTSF AVMVFTAFSS MLGQHKKQAV DWKTVFTMMP GMIFGVFTGA 101 LSAKYIPAFG LQIFFILFLT AVAFKTLHTD PQTASRPLPG LPGLTAVSTL 151 FGTMSSWVGI GGGSLSVPFL IHCGFPAHKA IGTSSGLAWP IALSGAISYL 201 LNGLNIAGLP EGSLGFLYLP AVAVLSAATI AFAPLGVKTA HKLSSAKLKK 251 XFGIMLLLIA GKMLYNLL* Computer analysis of this amino acid sequence gave the following results: Homology with hypothetical H.influenzae transmembrane protein HI0902 (accession number P44070) ORF17 and HI0902 proteins show 28% aa identity in 192 aa overlap: ORF17 3 HKKQAVNGKTVFTMMPGMIFGVFT-GAFSAKYIPAFGLQIF--FILFLTAVAFKTLHTDP 59 HK + + V + P ++ VF G F + +IF +++L ++ D HI0902 72 HKLGNIVWQAVRILAPVIMLSVFICGLFIGRLDREISAKIFACLWYLATKMVLSIKKD- 130 ORF17 60 QTAΞRPLPGLPXLTAVSTLFGTMSSWVGIGGGSLSVPFLIHCGFPAHKAIGTSSGLAWPI 119 Q ++ L L + L G SS GIGGG VPFL G +AIG+S+ + HI0902 131 QVTTKSLTPLSSVIG-GILIGMASSAAGIGGGGFIVPFLTARGINIKQAIGSSAFCGMLL 189 ORF17 120 ALSGAISYLLNGLNIAGLPEGSLGFLYLPAVAVLSAATIAFAPLGVXXXXXXXXXXXXXX 179 +SG S++++G +PE SLG++YLPAV ++A + + LG HI0902 190 GISGMFSFIVSGWGNPLMPEYSLGYIYLPAVLGITATSFFTSKLGASATAKLPVSTLKKG 249 ORF17 180 FGIMLLLIAGKM 191 F + L+++A M HI0902 250 FALFLIWAINM 261 Homology with a predicted ORF from N. meningitidis (strain A) ORF17 shows 96.9% identity over a 196aa overlap with an ORF (ORF17a) from strain A of N meningitidis: 10 20 30 orf17.pep GQHKKQAVΝGKTVFTMMPGMIFGVFTGAFS I I I II I II : I I II II I II I : II I I : I I : I orf17a QGLAQHPYAQHLAVGTSFAVMVFTAFSSMLGQHKKQAVDWKTVFTMMPGMVFGVFAGALS 50 60 70 80 90 100 40 50 60 70 80 90 orf17. pep AKYIPAFGLQIFFILFLTAVAFKTLHTDPQTASRPLPGLPXLTAVSTLFGTMSSWVGIGG II I I II II II I I II I I I I I I I I I I I II I M II II I I I II I I II I II I I I II I II I I I I I orf17a AKYIPAFGLQIFFILFLTAVAFKTLHTDPQTASRPLPGLPGLTAVSTLFGTMSSWVGIGG 110 120 130 140 150 160 100 110 120 130 140 150 orf17. pep GSLSVPFLIHCGFPAHKAIGTSSGLAWPIALSGAISYLLNGLNIAGLPEGSLGFLYLPAV I II II I I I II II I I II II I II I II II I II II I II I II I I I I I II II II I I I II I I II I II orf17a GSLSVPFLIHCGFPAHKAIGTSSGLAWPIALSGAISYLLNGLNIAGLPEGSLGFLYLPAV 170 180 190 200 210 220 160 170 180 190 orf17. pep AVLSAATIAFAPLGVKTAHKLSSAKLKKSFGIMLLLIAGKMLYNLLX I I I I I I I I II II I I I II I II II I I I II I II II II I I II II I I I II I I orfl7a AVLSAATIAFAPLGVKTAHKLSSAKLKKSFGIMLLLIAGKMLYNLLX 230 240 250 260 The complete length ORF 17a nucleotide sequence <SEQ ID 89> is: 1 ATGTGGCATT GGGACATTAT CTTAATCCTG CTTGCCGTAG GCAGTGCGGC 51 AGGTTTTATT GCCGGCCTGT TCGGCGTAGG CGGCGGCACG CTGATTGTCC 101 CTGTCGTTTT ATGGGTGCTT GATTTGCAGG GTTTGGCACA ACATCCTTAC 151 GCGCAACACC TCGCCGTCGG CACATCCTTC GCCGTCATGG TCTTCACCGC 201 CTTTTCCAGT ATGCTGGGGC AGCACAAAAA ACAGGCGGTC GACTGGAAAA 251 CCGTATTTAC GATGATGCCG GGTATGGTAT TCGGCGTATT CGCTGGCGCA 301 CTCTCCGCAA AATATATCCC AGCGTTCGGG CTTCAAATTT TCTTCATCCT 351 GTTTTTAACC GCCGTCGCAT TCAAAACACT GCATACCGAC CCTCAGACGG 401 CATCCCGCCC GCTGCCCGGA CTGCCCGGAC TGACTGCGGT TTCCACACTG 451 TTCGGCACAA TGTCGAGCTG GGTCGGCATA GGCGGCGGTT CACTTTCCGT 501 CCCCTTCTTA ATCCACTGCG GCTTCCCCGC CCATAAAGCC ATCGGCACAT 551 CATCCGGCCT TGCCTGGCCG ATTGCACTCT CCGGCGCAAT ATCGTATCTG 601 CTCAACGGCC TGAATATTGC AGGATTGCCC GAAGGGTCAC TGGGCTTCCT 651 TTACCTGCCC GCCGTCGCCG TCCTCAGCGC GGCAACCATT GCCTTTGCCC 701 CGCTCGGTGT CAAAACCGCC CACAAACTTT CTTCTGCCAA ACTCAAAAAA 751 TCCTTCGGCA TTATGTTGCT TTTGATTGCC GGAAAAATGC TGTACAACCT 801 GCTTTAA This encodes a protein having amino acid sequence <SEQ LD 90>: 1 MWHWDIILIL LAVGSAAGFI AGLFGVGGGT LIVPWLWVL DLQGLAQHPY 51 AQHLAVGTSF AVMVFTAFSS MLGQHKKQAV DWKTVFTMMP GMVFGVFAGA 101 LSAKYIPAFG LQIFFILFLT AVAFKTLHTD PQTASRPLPG LPGLTAVSTL 151 FGTMSSWVGI GGGSLSVPFL IHCGFPAHKA IGTSSGLAWP IALSGAISYL 201 LNGLNIAGLP EGSLGFLYLP AVAVLSAATI AFAPLGVKTA HKLSSAKLKK 251 SFGIMLLLIA GKMLYNLL* ORF17a and ORF17-1 show 98.9% identity in 268 aa overlap: 10 20 30 40 50 60 orf17a. pep MWHWDIILILLAVGSAAGFIAGLFGVGGGTLIVPWLWVLDLQGLAQHPYAQHLAVGTSF I II I I I II II II I I I I II II I II I II I I I II I I I I I I I II II I I II II I I II II I I II I I orf17-1 MWHWDIILILLAVGSAAGFIAGLFGVGGGTLIVPWLWVLDLQGLAQHPYAQHLAVGTSF 10 20 30 40 50 60 70 80 90 100 110 120 orf17 . pep AVMVFTAFSSMLGQHKKQAVDWKTVFTMMPGMVFGVFAGALSAKYIPAFGLQIFFILFLT I I I I I I I I II I I I I I II II II I II I II I II II : II I M I II II I I II I II I I II I I I I II orf17-1 AVMVFTAFSSMLGQHKKQAVDWKTVFTMMPGMIFGVFTGALSAKYIPAFGLQIFFILFLT 70 80 90 100 110 120 130 140 150 160 170 180 orf17a. pep AVAFKTLHTDPQTASRPLPGLPGLTAVSTLFGTMSSWVGIGGGSLSVPFLIHCGFPAHKA II II I I I I I I II I II I I I I I I I I II I II I II II I I I I I II II II I I I II I II I II I II II orf17-1 AVAFKTLHTDPQTASRPLPGLPGLTAVSTLFGTMSSWVGIGGGSLSVPFLIHCGFPAHKA 130 140 150 160 170 180 190 200 210 220 230 240 orf17a.pep IGTSSGLAWPIALSGAISYLLNGLNIAGLPEGSLGFLYLPAVAVLSAATIAFAPLGVKTA I I II I I I I I I I I I II I I I I I I II I II I I I II I II I II I I I II I II II I I II II I I I I I I I orf17-1 IGTSSGLAWPIALSGAISYLLNGLNIAGLPEGSLGFLYLPAVAVLSAATIAFAPLGVKTA 190 200 210 220 230 240 250 260 269 orf17a. pep HKLSSAKLKKSFGIMLLLIAGKMLYNLLX I II I I I II II I II I I I I I II I I I I II I I orfl7-l HKLSSAKLKKXFGIMLLLIAGKMLYNLLX 250 260 Homology with a predicted ORF from N.sonorrhoeae ORF 17 shows 93.9% identity over a 196aa overlap with a predicted ORF (ORF17.ng) from N gonorrhoeae: orf17.pep GQHKKQAVΝGKTVFTMMPGMIFGVFTGAFS 30 I II II II I : I I : I : I I I II I I I IM I I : I orfl7ng QGLAQHPYAQHLAVGTSFAVMVFTAFSSMLGQHKKQAVDWKTIFAMMPGMIFGVFAGALS 102 orf17.pep AKYIPAFGLQIFFILFLTAVAFKTLHTDPQTASRPLPGLPXLTAVSTLFGTMSSWVGIGG 90 I II II I I I II I I I I I I I II I I I I I II I I II II I I I II I I I I II II I M II II I I I I I orf17ng AKYIPAFGLQIFFILFLTAVAFKTLHTGRQTASRPLPGLPGLTAVSTLFGAMSSWVGIGG 162 orf17.pep GSLSVPFLIHCGFPAHKAIGTSSGLAWPIALSGAISYLLΝGLΝIAGLPEGΞLGFLYLPAV 150 I I II II I I I I I I I I II II II II I II I I I I I II II I I I M I I I I II I II I I I I I I I I I I I I orf17ng GSLSVPFLIHCGFPAHKAIGTSSGLAWPIALSGAISYLVΝGLΝIAGLPEGSLGFLYLPAV 202 orf17.pep AVLSAATIAFAPLGVKTAHKLSSAKLKKSFGIMLLLIAGKMLYΝLL 196 II I II I I I I I I II II II I I I I I I II II : II I I I I II II II I I I I II orf17ng AVLSAATIAFAPLGVKTAHKLSSAKLKESFGIMLLLIAGKMLYΝLL 268 An ORF17ng nucleotide sequence <SEQ LD 91> is predicted to encode a protein having amino acid sequence <SEQ ID 92>: 1 MWHWDIILIL LAVGSAAGFI AGLFGVGGGT LIVPWLWVL DLQGLAQHPY 51 AQHLAVGTSF AVMVFTAFSS MLGQHKKQAV DWKTIFAMMP GMIFGVFAGA 101 LSAKYIPAFG LQIFFILFLT AVAFKTLHTG RQTASRPLPG LPGLTAVSTL 151 FGAMSSWVGI GGGSLSVPFL IHCGFPAHKA IGTSSGLAWP IALSGAISYL 201 VΝGLΝIAGLP EGSLGFLYLP AVAVLSAATI AFAPLGVKTA HKLSSAKLKE 251 SFGIMLLLIA GKMLYΝLL* Further work revealed the complete gonococcal DΝA sequence <SEQ ID 93>: 1 ATGTGGCATT GGGACATTAT CTTAATCCTG CTTGCcgtag gcAGTGCGGC 51 AGGTTTTATT GCCGGCCTGT Tcggtgtagg cggcgGTACG CTGATTGTCC 101 CTGTCGTTTT ATGGGTGCTT GATTTGCAGG GTTTGGCACA ACATCCTTAC 151 GCGCAACACC TCGCCGTCGG CAcaTccttc gcCGTCATGG TCTTCACCGC 201 CTTTTCCAGT ATGTTGGGGC AGCACAAAAA ACAGGCGGTC GACTGGAAAA 251 CCATATTTGC GATGATGCCG GGTATGATAT TCGGCGTATT CGCTGGCGCA 301 CTCTCCGCAA AATATATCCC CGCGTTCGGG CTTCAAATTT TCTTCATCCT 351 GTTTTTAACC GCCGTCGCAT TCAAAACACT GCATACCGGT CGTCAGACGG 401 CATCCCGCCC GCTGCCCGGG CTGCCCGGAC TGACTGCGGT TTCCACACTG 451 TTCGGCGCAA TGTCGAGCTG GGTCGGCATA GGCGGCGGTT CACTTTCCGT 501 CCCCTTCTTA ATCCACTGCG GCTTCCCCGC CCATAAAGCC ATCGGCACAT 551 CATCCGGCCT TGCCTGGCCG ATTGCACTCT CCGGCGCAAT ATCGTATCTG 601 GTCAACGGTC TGAATATTGC AGGATTGCCC GAAGGGTCGC TGGGCTTCCT 651 TTACCTGCCC GCCGTCGCCG TCCTCAGCGC GGCAACCATT GCCTTTGCCC 701 CGCTCGGTGT CAAAACCGCC CACAAACTTT CTTCTGCCAA ACTCAAAGAA 751 TCCTTCGGCA TTATGTTGCT TTTGATTGCC GGAAAAATGC TGTACAACCT 801 GCTTTAA This corresponds to the amino acid sequence <SEQ ID 94; ORF17ng-l>: 1 MWHWDIILIL LAVGSAAGFI AGLFGVGGGT LIVPWLWVL DLQGLAQHPY 51 AQHLAVGTSF AVMVFTAFSS MLGQHKKQAV DWKTIFAMMP GMIFGVFAGA 101 LSAKYIPAFG LQIFFILFLT AVAFKTLHTG RQTASRPLPG LPGLTAVSTL 151 FGAMSSWVGI GGGSLSVPFL IHCGFPAHKA IGTSSGLAWP IALSGAISYL 201 VΝGLΝIAGLP EGSLGFLYLP AVAVLSAATI AFAPLGVKTA HKLSSAKLKE 251 SFGIMLLLIA GKMLYΝLL* ORF17ng-l and ORF17-1 show 96.6% identity in 268 aa overlap: 10 20 30 40 50 60 orfl7-l.pep MWHWDIILILLAVGSAAGFIAGLFGVGGGTLIVPWLWVLDLQGLAQHPYAQHLAVGTSF II I I II I II II I I I I I I II I II II I II I II I I II II I I II I I I II II I I I I II II I I II I orf 17ng-l MWHWDIILILLAVGSAAGFIAGLFGVGGGTLIVPWLWVLDLQGLAQHPYAQHLAVGTSF 10 20 30 40 50 60 70 80 90 100 110 120 orf 17-1. pep AVMVFTAFSSMLGQHKKQAVDWKTVFTMMPGMIFGVFTGALSAKYIPAFGLQIFFILFLT I I II II I I II I II II I II II I II I : I: I I I I II I I I I : I II I II II I I I II I I I I I II I I orfl7ng-l AVMVFTAFSSMLGQHKKQAVDWKTIFAMMPGMIFGVFAGALSAKYIPAFGLQIFFILFLT 70 80 90 100 110 120 130 140 150 160 170 180 orf 17-1. pep AVAFKTLHTDPQTASRPLPGLPGLTAVSTLFGTMSSWVGIGGGSLSVPFLIHCGFPAHKA I II II I II I I I I I II II I II I II I II II II : II II II I II II II I I I I I I I I I I II I I orfl7ng-l AVAFKTLHTGRQTASRPLPGLPGLTAVSTLFGAMSSWVGIGGGSLSVPFLIHCGFPAHKA 130 140 150 160 170 180 190 200 210 220 230 240 orf 17-1 . pep IGTSSGLAWPIALSGAISYLLNGLNIAGLPEGSLGFLYLPAVAVLSAATIAFAPLGVKTA I I I I I I I II II I I II I I I IM II I II I I I II II I II II I I I I I II II I I I I I II II I I I I orfl7ng-l IGTSSGLAWPIALSGAISYLVNGLNIAGLPEGSLGFLYLPAVAVLSAATIAFAPLGVKTA 190 200 210 220 230 240 250 260 269 orf17-1.pep HKLSSAKLKKXFGIMLLLIAGKMLYNLLX I I I I II I II : II I I I I I I II I I I I I II I orf17ng-l HKLSSAKLKESFGIMLLLIAGKMLYNLLX 250 260 In addition, ORF17ng-l shows significant homology with a hypothetical H.influenzae protein: sp|P44070 |Y902_HAEIN HYPOTHETICAL PROTEIN HI0902 pir||G64015 hypothetical protein HI0902 - Haemophilus influenzae (strain Rd KW20) gi 11573922 (U32772) H. influenzae predicted coding region HI0902 [Haemophilus influenzae] Length = 264 Score = 74 (34.9 bits), Expect = 1.6e-23, Sum P(2) = 1.6e-23 Identities = 15/43 (34%), Positives = 23/43 (53%) Query: 55 AVGTSFAVMVFTAFSSMLGQHKKQAVDWKTIFAMMPGMIFGVF 97 A+GTSFA +V T S HK + W+ + + P ++ VF Sbjct: 52 ALGTSFATIVITGIGSAQRHHKLGNIVWQAVRILAPVIMLSVF 94 Score = 195 (91.9 bits), Expect = 1.6e-23, Sum P(2) = 1.6e-23 Identities = 44/114 (38%), Positives = 65/114 (57%) Query: 150 LFGAMSSWVGIGGGSLSVPFLIHCGFPAHKAIGTSSGLAWPIALSGAISYLVNGLNIAGL 209 L G SS GIGGG VPFL G +AIG+S+ + +SG S++V+G + Sbjct: 148 LIGMASSAAGIGGGGFIVPFLTARGINIKQAIGSSAFCGMLLGISGMFSFIVSGWGNPLM 207 Query: 210 PEGSLGFLYLPAVAVLSAATIAFAPLGVKTAHKLSSAKLKESFGIMLLLIAGKM 263 PE SLG++YLPAV ++A + + LG KL + LK+ F + L+++A M Sbjct: 208 PEYSLGYIYLPAVLGITATSFFTSKLGASATAKLPVSTLKKGFALFLIWAINM 261 This .analysis, including the homology with the hypothetical H.influenzae transmembrane protein, suggests that the proteins from N. meningitidis and N.gonorrhoeae, and their epitopes, could be useful antigens for vaccines or diagnostics, or for raising antibodies. Example 12 The following partial DNA sequence was identified in N. meningitidis <SEQ ID 95>: 1 .. GGAAACGGAT GGCAGGCAGA CCCCGAACAT CCGCTGCTCG GGCTTTTTGC 51 CGTCAGTAAT GTATCGATGA CGCTTGCTTT TGTCGGAATA TGTGCGTTGG 101 TGCATTATTG CTTTTCGGGA ACGGTTCAAG TGTTTGTGTT TGCGGCACTG 151 CTCAAACTTT ATGCGCTGAA GCCGGTTTAT TGGTTCGTGT TGCAGTTTGT 201 GCTGATGGCG GTTGCCTATG TCCACCGCTG CGGTATAGAC CGGCAGCCGC 251 CGTCAACGTT CGGCGGCTCG CAGCTGCGAC TCGGCGGGTT GACGGCAGCG 301 TTGATGCAGG TCTCGGTACT GGTGCTGCTG CTTTCAGAAA TTGGAAGATA 351 A This corresponds to the amino acid sequence <SEQ ID 96; ORF18>: 1 .. GNGWQADPEH PLLGLFAVSN VSMTLAFVGI CALVHYCFSG TVQVFVFAAL 51 LKLYALKPVY WFVLQFVLMA VAYVHRCGID RQPPSTFGGS QLRLGGLTAA 101 LMQVSVLVLL LSEIGR* Further work revealed the complete nucleotide sequence <SEQ ID 97>: 1 ATGATTTTGC TGCATTTGGA TTTTTTGTCT GCCTTACTGT ATGCGGCGGT 51 TTTTCTGTTT CTGATATTCC GCGCAGGAAT GTTGCAATGG TTTTGGGCGA 101 GTATTATGCT GTGGCTGGGC ATATCGGTTT TGGGGGCAAA GCTGATGCCC 151 GGCATATGGG GAATGACCCG CGCCGCGCCC TTGTTCATCC CCCATTTTTA 201 CCTGACTTTG GGCAGCATAT TTTTTTTCAT CGGGCATTGG AACCGGAAAA 251 CAGATGGAAA CGGATGGCAG GCAGACCCCG AACATCCGCT GCTCGGGCTT 301 TTTGCCGTCA GTAATGTATC GATGACGCTT GCTTTTGTCG GAATATGTGC 351 GTTGGTGCAT TATTGCTTTT CGGGAACGGT TCAAGTGTTT GTGTTTGCGG 401 CACTGCTCAA ACTTTATGCG CTGAAGCCGG TTTATTGGTT CGTGTTGCAG 451 TTTGTGCTGA TGGCGGTTGC CTATGTCCAC CGCTGCGGTA TAGACCGGCA 501 GCCGCCGTCA ACGTTCGGCG GCTCGCAGCT GCGACTCGGC GGGTTGACGG 551 CAGCGTTGAT GCAGGTCTCG GTACTGGTGC TGCTGCTTTC AGAAATTGGA 601 AGATAA This corresponds to the amino acid sequence <SEQ ID 98; ORF18-l>: 1 MILLHLDFLS ALLYAAVFLF LIFRAGMLQW FWASIMLWLG ISVLGAKLMP 51 GIWGMTRAAP LFIPHFYLTL GSIFFFIGHW NRKTDGNGWQ ADPEHPLLGL 101 FAVSNVSMTL AFVGICALVH YCFSGTVQVF VFAALLKLYA LKPVYWFVLQ 151 FVLMAVAYVH RCGIDRQPPS TFGGSQLRLG GLTAALMQVS VLVLLLSEIG 201 R* Computer analysis of this amino acid sequence gave the following results: Homology with a predicted ORF from N.meninsitidis (strain Al ORF18 shows 98.3% identity over a 116aa overlap with an ORF (ORF18a) from strain A of N meningitidis: 10 20 30 orf 18 . pep GΝGWQADPEHPLLGLFAVSΝVSMTLAFVGI I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I orflδa TRAAPLFIPHFYLTLGSIFFFIGHWΝRKTDGΝGWQADPEHPLLGLFAVSΝVSMTLAFVGI 60 70 80 90 100 110 40 50 60 70 80 90 orf 18 . pep CALVHYCFSGTVQVFVFAALLKLYALKPVYWFVLQFVLMAVAYVHRCGIDRQPPSTFGGS I I I II I II I II I I I I I I I I I I I I I I II II I II I I I I II I II I I II I I I I I I II I I I I I I orfl 8a CALVHYCFSXTVQVFVFAALLKLYALKPVYWFVLQFVLMAVAYVHRCGIDRQPPSTFGGS 120 130 140 150 160 170 100 110 orf 18 . pep QLRLGGLTAALMQVSVLVLLLSEIGRX II I II I II II II I I II I I I I I I II I I orflδa QLRLGGLTAALMQXSVLVLLLSEIGRX 180 190 200 The complete length ORF 18a nucleotide sequence <SEQ LD 99> is: 1 ATGATTTTGC TGCATTTGGA TTTTTTGTCT GCCTTACTGT ATGCGGCGGT 51 TTTTCTGTTT CTGATATTCC GCGCAGGAAT GTTGCAATGG TTTTGGGCGA 101 GTATTATGCT GTGGCTGGGC ATATCGGTTT TGGGGGCAAA GCTGATGCCC 151 GGCATATGGG GAATGACCCG CGCCGCGCCC TTGTTCATCC CCCATTTTTA 201 CCTGACTTTG GGCAGCATAT TTTTTTTCAT CGGGCATTGG AACCGGAAAA 251 CGGATGGAAA CGGATGGCAG GCAGACCCCG AACATCCTCT GCTCGGGCTG 301 TTTGCCGTCA GTAATGTATC GATGACGCTT GCTTTTGTCG GAATATGTGC 351 GTTGGTGCAT TATTGCTTTT CGΝGAACGGT TCAAGTGTTT GTGTTTGCGG 401 CACTGCTCAA ACTTTATGCG CTGAAGCCGG TTTATTGGTT CGTGTTGCAG 451 TTTGTGCTGA TGGCGGTTGC CTATGTCCAC CGCTGCGGTA TAGACCGGCA 501 GCCGCCGTCA ACGTTCGGCG GNTCGCAGCT GCGACTCGGC GGGTTGACGG 551 CAGCGTTGAT GCAGNTCTCG GTACTGGTGC TGCTGCTTTC AGAAATTGGA 601 AGATAA This encodes a protein having amino acid sequence <SEQ ID 100>: 1 MILLHLDFLS ALLYAAVFLF LIFRAGMLQW FWASIMLWLG ISVLGAKLMP 51 GIWGMTRAAP LFIPHFYLTL GSIFFFIGHW NRKTDGNGWQ ADPEHPLLGL 101 FAVSNVSMTL AFVGICALVH YCFSXTVQVF VFAALLKLYA LKPVYWFVLQ 151 FVLMAVAYVH RCGIDRQPPS TFGGSQLRLG GLTAALMQXS VLVLLLSEIG 201 R* ORFl 8a and ORFl 8-1 show 99.0% identity in 201 aa overlap: 10 20 30 40 50 60 orf18a. pep MILLHLDFLSALLYAAVFLFLIFRAGMLQWFWASIMLWLGISVLGAKLMPGIWGMTRAAP I I II I I II I I I II II II II I II I I I I I I II II I I II II I II I I I I II I I I II II II II I I orf18-1 MILLHLDFLSALLYAAVFLFLIFRAGMLQWFWASIMLWLGISVLGAKLMPGIWGMTRAAP 10 20 30 40 50 60 70 80 90 100 110 120 orf18a. pep LFIPHFYLTLGSIFFFIGHWNRKTDGNGWQADPEHPLLGLFAVSNVSMTLAFVGICALVH I I II I I II I II I II I I I II I I II II I I I I II II II II I II I II II I II I I II II I II I I I orf18-1 LFIPHFYLTLGSIFFFIGHWNRKTDGNGWQADPEHPLLGLFAVSNVSMTLAFVGICALVH 70 80 90 100 110 120 130 140 150 160 170 180 orf18a. pep YCFSXTVQVFVFAALLKLYALKPVYWFVLQFVLMAVAYVHRCGIDRQPPSTFGGSQLRLG II I I I I II II I I I II I I II I I I I II I II II I I II II II I I I I I I II I I I II II II II I I orf18-1 YCFSGTVQVFVFAALLKLYALKPVYWFVLQFVLMAVAYVHRCGIDRQPPSTFGGSQLRLG 130 140 150 160 170 180 190 200 orf18a. pep GLTAALMQXSVLVLLLSEIGRX I II II I I I I II I I I I II I I II orfl8-l GLTAALMQVSVLVLLLSEIGRX 190 200 Homology with a predicted ORF from N.gonorrhoeae ORF 18 shows 93.1% identity over a 116aa overlap with a predicted ORF (ORF18.ng) from N. gonorrhoeae: orf18. pep GNGWQADPEHPLLGLFAVSNVSMTLAFVGI 30 II II I I II II I I I I II II II I I I I I I I I I I orflδng TRAAPLFIPHFYLTLGSIFFFIGYWNRKTDGNGWQADPEHPLLGLFAVSNVSMTLAFVGI 115 orf18.pep CALVHYCFSGTVQVFVFAALLKLYALKPVYWFVLQFVLMAVAYVHRCGIDRQPPSTFGGS 90 II I II I II I I II I II I II I I II I I II I II I II II II II I I I II I II II II I I II I I I I I I orflδng CALVHYCFSGTVQVFVFAALLKLYALKPVYWFVLQFVLMAVAYVHRCGIDRQPPSTFGGS 175 orf18.pep QLRLGGLTAALMQVSVLVLLLSEIGR 116 I II I I Ml I II I : I : : II : I II I orf18ng QLRLGVLAAMLMQVAVTAMLLAEIGR 201 The complete length ORFlδng nucleotide sequence is <SEQ ID 101>: 1 ATGATTTTGC TGCATTTGGA TTTTTTGTCT GCCTTACTGt aTGCGGcggt 51 tttTctgTTT CTGATATTCC GCGCAGGAAT GTTGCAATGG TTTTGGGCGA 101 GTATTGCGTT GTGGCTCGGC ATCTCGGTTT TAGGGGTAAA GCTGATGCCG 151 GGGATGTGGG GAATGACCCG CGCCGCGCCT TTGTTCATCC CCCATTTTTA 201 CCTGACTTTG GGCAGCATAT TTTTTTTCAT CGGGTATTGG AACCGGAAAA 251 CAGATGGAAA CGGATGGCAG GCAGACCCCG AACATCCGCT GCTCGGGCTT 301 TTTGCCGTCA GTAATGTATC GATGACGCTT GCTTTTGTCG GAATATGTGC 351 GTTGGTGCAT TATTGCTTTT CGGGAACGGT TCAAGTGTTT GTGTTTGCGG 401 CATTGCTCAA ACTTTATGCG CTGAAGCCGG TTTATTGGTT CGTGTTGCAG 451 TTTGTATTGA TGGCGGttgC CTATGTCCAC CGCTGCGGTA TAGACCGGCA 501 GCCGCCGTCA ACGTTCGGCG GTTCGCAGCT GCGACTCGGC GTGTTGGCGG 551 CGATGTTGAT GCAGGTTGCG GTAACGGCGA TGCTGCTTGC CGAAATCGGC 601 AGATGA This encodes a protein having amino acid sequence <SEQ LD 102>: 1 MILLHLDFLS ALLYAAVFLF LIFRAGMLQW FWASIALWLG ISVLGVKLMP 51 GMWGMTRAAP LFIPHFYLTL GSIFFFIGYW NRKTDGNGWQ ADPEHPLLGL 101 FAVSNVSMTL AFVGICALVH YCFSGTVQVF VFAALLKLYA LKPVYWFVLQ 151 FVLMAVAYVH RCGIDRQPPS TFGGSQLRLG VLAAMLMQVA VTAMLLAEIG 201 R* This ORF18ng protein sequence shows 94.0% identity in 201 aa overlap with ORFl 8-1: 10 20 30 40 50 60 orf18-1. pep MILLHLDFLSALLYAAVFLFLIFRAGMLQWFWASIMLWLGISVLGAKLMPGIWGMTRAAP I I I II I I I I II I I I II II II I I II I II I II II I I I I I II I II I I : I I I I I : II I II I I I orflδng MILLHLDFLSALLYAAVFLFLIFRAGMLQWFWASIALWLGISVLGVKLMPGMWGMTRAAP 10 20 30 40 50 60 70 80 90 100 110 120 orf18-1. pep LFIPHFYLTLGSIFFFIGHWNRKTDGNGWQADPEHPLLGLFAVSNVSMTLAFVGICALVH I II II II II II I I II II I : I I II II I I II I I I I II II I I II I II II II I I II I I I II I II orflδng LFIPHFYLTLGSIFFFIGYWNRKTDGNGWQADPEHPLLGLFAVSNVSMTLAFVGICALVH 70 80 90 100 110 120 130 140 150 160 170 180 orf18-1. pep YCFSGTVQVFVFAALLKLYALKPVYWFVLQFVLMAVAYVHRCGIDRQPPSTFGGSQLRLG I II II II II II II I I II I II I I II I II I II II I I I I I I II II I I I II I II I II I I I I II I orfl8ng YCFSGTVQVFVFAALLKLYALKPVYWFVLQFVLMAVAYVHRCGIDRQPPSTFGGSQLRLG 130 140 150 160 170 180 190 200 orf18-1. pep GLTAALMQVSVLVLLLSEIGRX I : I I II I: I :: II : I I II I orflδng VLAAMLMQVAVTAMLLAEIGRX 190 200 Based on this analysis, including the presence of several putative transmembrane domains in the gonococcal protein, it is predicted that the proteins from N. meningitidis and N.gonorrhoeae, and their epitopes, could be useful antigens for vaccines or diagnostics, or for raising antibodies. Example 13 The following partial DNA sequence was identified in N. meningitidis <SEQ ID 103>: 1 ATGAAAACCC CACTCCTCAA GCCTCTGCTN ATTACCTCGC TTCCCGTTTT 51 CGCCAGTGTT TTTACCGCCG CCTCCATCGT CTGGCAGCTA GGCGAACCCA 101 AGCTCGCCAT GCCCTTCGTA CTCGGCATCA TCGCCGGCGG CCTTGTCGAT 151 TTGGACAACC NCNTGACCGG ACGGCTNAAA AACATCATCA CCACCGTCGC 201 CCTGTTCACC CTCTCCTCGC TCACGGCACA AAGCACCCTC GGCACAGGGC 251 TGCCCTTCAT CCTCGCCATG ACCCTGATGA CTT. CG. CTT CACCATTTTA 301 GGCGCGGNCG ... This corresponds to the amino acid sequence <SEQ ID 104; ORF19>: 1 MKTPLLKPLL ITSLPVFASV FTAASIVWQL GEPKLAMPFV LGIIAGGLVD 51 LDNXXTGRLK NIITTVALFT LSSLTAQSTL GTGLPFILAM TLMTXXFTIL 101 GAX... Further work revealed the complete nucleotide sequence <SEQ ID 105>: 1 ATGAAAACCC CACTCCTCAA GCCTCTGCTC ATTACCTCGC TTCCCGTTTT 51 CGCCAGTGTT TTTACCGCCG CCTCCATCGT CTGGCAGCTA GGCGAACCCA 101 AGCTCGCCAT GCCCTTCGTA CTCGGCATCA TCGCCGGCGG CCTTGTCGAT 151 TTGGACAACC GCCTGACCGG ACGGCTGAAA AACATCATCA CCACCGTCGC 201 CCTGTTCACC CTCTCCTCGC TCACGGCACA AAGCACCCTC GGCACAGGGC 251 TGCCCTTCAT CCTCGCCATG ACCCTGATGA CCTTCGGCTT CACCATTTTA 301 GGCGCGGTCG GGCTCAAATA CCGCACCTTC GCCTTCGGTG CACTCGCCGT 351 CGCCACCTAC ACCACACTTA CCTACACCCC CGAAACCTAC TGGCTGACCA 401 ACCCCTTCAT GATTTTATGC GGCACCGTAC TGTACAGCAC CGCCATCCTC 451 CTGTTCCAAA TCGTCCTGCC CCACCGCCCC GTCCAAGAAA GCGTCGCCAA 501 CGCCTACGAC GCACTCGGCG GCTACCTCGA AGCCAAAGCC GACTTCTTCG 551 ACCCCGATGA GGCAGCCTGG ATAGGCAACC GCCACATCGA CCTCGCCATG 601 AGCAACACCG GCGTCATCAC CGCCTTCAAC CAATGCCGTT CCGCCCTGTT 651 TTACCGCCTT CGCGGCAAAC ACCGCCACCC GCGCACCGCC AAAATGCTGC 701 GTTACTACTT TGCCGCCCAA GACATACACG AACGCATCAG CTCCGCCCAC 751 GTCGATTATC AGGAAATGTC CGAAAAATTC AAAAACACCG ACATCATCTT 801 CCGCATCCAC CGCCTGCTCG AAATGCAGGG ACAAGCCTGC CGCAACACCG 851 CCCAAGCCCT GCGCGCAAGC AAAGACTACG TTTACAGCAA ACGCCTCGGC 901 CGCGCCATCG AAGGCTGCCG CCAATCGCTG CGCCTCCTTT CAGACAGCAA 951 CGACAGTCCC GACATCCGCC ACCTGCGCCG CCTTCTCGAC AACCTCGGCA 1001 GCGTCGACCA GCAGTTCCGC CAACTCCAGC ACAACGGCCT GCAGGCAGAA 1051 AACGACCGCA TGGGCGACAC CCGCATCGCC GCCCTCGAAA CCAGCAGCCT 1101 CAAAAACACC TGGCAGGCAA TCCGTCCGCA GCTAAACCTC GAATCAGGCG 1151 TATTCCGCCA TGCCGTCCGC CTGTCCCTCG TCGTTGCCGC CGCCTGCACC 1201 ATCGTCGAAG CCCTCAACCT CAACCTCGGC TACTGGATAC TACTGACCGC 1251 CCTTTTCGTC TGCCAACCCA ACTACACCGC CACCAAAAGC CGCGTCCGCC 1301 AGCGCATCGC CGGCACCGTA CTCGGCGTAA TCGTCGGCTC GCTCGTCCCC 1351 TACTTCACCC CGTCTGTCGA AACCAAACTC TGGATTGTCA TCGCCAGTAC 1401 CACCCTCTTT TTCATGACCC GCACCTACAA ATACAGTTTC TCCACCTTCT 1451 TCATTACCAT TCAAGCCCTG ACCAGCCTCT CCCTCGCAGG TTTGGACGTA 1501 TACGCCGCCA TGCCCGTACG CATCATCGAC ACCATTATCG GCGCATCCCT 1551 TGCCTGGGCG GCAGTCAGCT ACCTGTGGCC AGACTGGAAA TACCTCACGC 1601 TCGAACGCAC CGCCGCCCTT GCCGTATGCA GCAACGGTGC CTATCTCGAA 1651 AAAATCACCG AACGCCTCAA AAGCGGCGAA ACCGGCGACG ACGTCGAATA 1701 CCGCGCCACC CGCCGCCGCG CCCACGAACA CACCGCCGCC CTCAGCAGCA 1751 CCCTTTCCGA CATGAGCAGC GAACCCGCAA AATTCGCCGA CAGCCTGCAA 1801 CCCGGCTTTA CCCTGCTCAA AACCGGCTAC GCCCTGACCG GCTACATCTC 1851 CGCCCTCGGC GCATACCGCA GCGAAATGCA CGAAGAATGC AGCCCCGACT 1901 TTACCGCACA GTTCCACCTC GCCGCCGAAC ACACCGCCCA CATCTTCCAA 1951 CACCTGCCCG AAACCGAACC CGACGACTTT CAGACAGCAC TGGATACACT 2001 GCGCGGCGAA CTCGACACCC TCCGCACCCA CAGCAGCGGA ACACAAAGCC 2051 ACATCCTCCT CCAACAGCTC CAACTCATCG CCCGACAGCT CGAACCCTAC 2101 TACCGCGCCT ACCGCCAAAT TCCGCACAGG CAGCCCCAAA ATGCAGCCTG 2151 A This corresponds to the amino acid sequence <SEQ ID 106; ORF19-l>: 1 MKTPLLKPLL ITSLPVFASV FTAASIVWQL GEPKLAMPFV LGIIAGGLVD 51 LDNRLTGRLK NIITTVALFT LSSLTAQSTL GTGLPFILAM TLMTFGFTIL 101 GAVGLKYRTF AFGALAVATY TTLTYTPETY WLTNPFMILC GTVLYSTAIL 151 LFQIVLPHRP VQESVANAYD ALGGYLEAKA DFFDPDEAAW IGNRHIDLAM 201 SNTGVITAFN QCRSALFYRL RGKHRHPRTA KMLRYYFAAQ DIHERISSAH 251 VDYQEMSEKF KNTDIIFRIH RLLEMQGQAC RNTAQALRAS KDYVYSKRLG 301 RAIEGCRQSL RLLSDSNDSP DIRHLRRLLD NLGSVDQQFR QLQHNGLQAE 351 NDRMGDTRIA ALETSSLKNT WQAIRPQLNL ESGVFRHAVR LSLWAAACT 401 IVEALNLNLG YWILLTALFV CQPNYTATKS RVRQRIAGTV LGVIVGSLVP 451 YFTPSVETKL WIVIASTTLF FMTRTYKYSF STFFITIQAL TSLSLAGLDV 501 YAAMPVRIID TIIGASLAWA AVSYLWPDWK YLTLERTAAL AVCSNGAYLE 551 KITERLKSGE TGDDVEYRAT RRRAHEHTAA LSSTLSDMSS EPAKFADSLQ 601 PGFTLLKTGY ALTGYISALG AYRSEMHEEC SPDFTAQFHL AAEHTAHIFQ 651 HLPETEPDDF QTALDTLRGE LDTLRTHSSG TQSHILLQQL QLIARQLEPY 701 YRAYRQIPHR QPQNAA* Computer analysis of this amino acid sequence gave the following results: Homology with predicted transmenbrane protein YHFK of H. influenzae (accession number P44289) ORF19 and YHFK proteins show 45% aa identity in 97 aa overlap: orfl9 6 LKPLLITSLPVFASVFTAASIVWQLGEPKLAMPFVLGIIAGGLVDLDNXXTGRLKNIITT 65 L +I+++PVF +V AA +W +MP +LGIIAGGLVDLDN TGRLKN+ T YHFK 5 LNAKVISTIPVFIAVNIAAVGIWFFDISSQSMPLILGIIAGGLVDLDNRLTGRLKNVFFT 64 orfl9 66 VALFTLSSLTAQSTLGTGLPFILAMTLMTXXFTILGA 102 + F++SS Q +G + +1+ MT++T FT++GA YHFK 65 LIAFSISSFIVQLHIGKPIQYIVLMTVLTFIFTMIGA 101 Homology with a predicted ORF from N.meninsitidis (strain A) ORF19 shows 92.2% identity over a 102aa overlap with an ORF (ORF19a) from strain A of N. meningitidis: 10 20 30 40 50 60 orf19. pep MKTPLLKPLLITSLPVFASVFTAASIVWQLGEPKLAMPFVLGIIAGGLVDLDΝXXTGRLK MM I II I I I I I I I I I I II I I I I I I I II I I II I I I I I I I II I I I II I I I II I II II I orfl9a MKTPPLKPLLITSLPVFASVFTAASIVWQLGEPKLAMPFVLGIIAGGLVDLDΝRLTGRLK 10 20 30 40 50 60 70 80 90 100 orf19. pep ΝIITTVALFTLSSLTAQSTLGTGLPFILAMTLMTXXFTILGAX I I I: I II I II I II I : I I II I I II I II I I I II I I I M M orf19a ΝIIATVALFTLSSLVAQSTLGTGLPFILAMTLMTFGFTIMGAVGLKYRTFAFGA AVATY 70 80 90 100 110 120 orf19a TTLTYTPETYWLTΝPFMILCGTVLYΞTAIILFQIILPHRPVQEΝVAΝAYEALGSYLEAKA 130 140 150 160 170 180 The complete length ORFl 9a nucleotide sequence <SEQ ID 107> is: 1 ATGAAAACCC CACCCCTCAA GCCTCTGCTC ATTACCTCGC TTCCCGTTTT 51 CGCCAGTGTC TTTACCGCCG CCTCCATCGT CTGGCAGCTG GGCGAACCCA 101 AGCTCGCCAT GCCCTTCGTA CTCGGCATCA TCGCTGGCGG CCTGGTCGAT 151 TTGGACAACC GCCTGACCGG ACGGCTGAAA AACATCATCG CCACCGTCGC 201 CCTGTTCACC CTCTCCTCAC TTGTCGCGCA AAGCACCCTC GGCACAGGTT 251 TGCCATTCAT CCTCGCCATG ACCCTGATGA CTTTCGGCTT TACCATCATG 301 GGCGCGGTCG GGCTGAAATA CCGCACCTTC GCCTTCGGCG CACTCGCCGT 351 CGCCACCTAC ACCACACTTA CCTACACCCC CGAAACCTAC TGGCTGACCA 401 ACCCCTTTAT GATTCTGTGC GGAACCGTAC TGTACAGCAC CGCCATCATC 451 CTGTTCCAAA TCATCCTGCC CCACCGCCCC GTTCAAGAAA ACGTCGCCAA 501 CGCCTACGAA GCACTCGGCA GCTACCTCGA AGCCAAAGCC GACTTTTTCG 551 ATCCCGACGA AGCCGAATGG ATAGGCAACC GCCACATCGA CCTCGCCATG 601 AGCAACACCG GCGTCATCAC CGCCTTCAAC CAATGCCGTT CCGCCCTGTT 651 TTACCGCCTT CGCGGCAAAC ACCGCCACCC GCGCACCGCC AAAATGCTGC 701 GCTACTACTT CGCCGCCCAA GACATACACG AACGCATCAG CTCCGCCCAC 751 GTCGACTACC AAGAGATGTC CGAAAAATTC AAAAACACCG ACATCATCTT 801 CCGCATCCAC CGCCTGCTCG AAATGCAGGG ACAAGCCTGC CGCAACACCG 851 CCCAAGCCCT GCGCGCAAGC AAAGACTACG TTTACAGCAA ACGCCTCGGC 901 CGCGCCATCG AAGGCTGCCG CCAATCGCTG CGCCTCCTTT CAGACAGCAA 951 CGACAATCCC GACATCCGCC ACCTGCGCCG CCTTCTCGAC AACCTCGGCA 1001 GCGTCGACCA GCAGTTCCGC CAACTCCAGC ACAACGGCCT GCAGGCAGAA 1051 AACGACCGCA TGGGCGACAC CCGCATCGCC GCCCTCGAAA CCGGCAGCCT 1101 CAAAAACACC TGGCAGGCAA TCCGTCCGCA GCTAAACCTC GAATCAGGCG 1151 TATTCCGCCA TGCCGTCCGC CTGTCCCTTG TCGTTGCCGC CGCCTGCACC 1201 ATCGTCGAAG CCCTCAACCT CAACCTCGGC TACTGGATAC TACTGACCGC 1251 CCTTTTCGTC TGCCAACCCA ACTACACCGC CACCAAAAGC CGCGTCCGCC 1301 AGCGCATCGC CGGCACCGTA CTCGGCGTAA TCGTCGGCTC GCTCGTCCCC 1351 TACTTTACCC CCTCCGTCGA AACCAAACTC TGGATCGTCA TCGCCAGTAC 1401 CACCCTCTTT TTCATGACCC GCACCTACAA ATACAGCTTC TCGACATTTT 1451 TCATCACCAT TCAAGCCCTG ACCAGCCTCT CCCTCGCAGG GTTGGACGTA 1501 TACGCCGCCA TGCCCGTACG CATCATCGAC ACCATTATCG GCGCATCCCT 1551 TGCCTGGGCG GCAGTCAGCT ACCTGTGGCC AGACTGGAAA TACCTCACGC 1601 TCGAACGCAC CGCCGCCCTT GCCGTATGCA GCAACGGCGC CTATCTCGAA 1651 AAAATCACCG AACGCCTCAA AAGCGGCGAA ACCGGCGACG ACGTCGAATA 1701 CCGCGCCACC CGCCGCCGCG CCCACGAACA CACCGCCGCC CTCAGCAGCA 1751 CCCTTTCCGA CATGAGCAGC GAACCCGCAA AATTCGCCGA CAGCCTGCAA 1801 CCCGGCTTTA CCCTGCTCAA AACCGGCTAC GCCCTGACCG GCTACATCTC 1851 CGCCCTCGGC GCATACCGCA GCGAAATGCA CGAAGAATGC AGCCCCGACT 1901 TTACCGCACA GTTCCACCTC GCCGCCGAAC ACACCGCCCA CATCTTCCAA 1951 CACCTGCCCG AAACCGAACC CGACGACTTT CAGACAGCAC TGGATACACT 2001 GCGCGGCGAA CTCGACACCC TCCGCACCCA CAGCAGCGGA ACACAAAGCC 2051 ACATCCTCCT CCAACAGCTC CAACTCATCG CCCGGCAGCT CGAACCCTAC 2101 TACCGCGCCT ACCGACAAAT TCCGCACAGG CAGCCCCAAA ACGCAGCCTG 2151 A This encodes a protein having amino acid sequence <SEQ LD 108>: 1 MKTPPLKPLL ITSLPVFASV FTAASIVWQL GEPKLAMPFV LGIIAGGLVD 51 LDNRLTGRLK NIIATVALFT LSSLVAQSTL GTGLPFILAM TLMTFGFTIM 101 GAVGLKYRTF AFGALAVATY TTLTYTPETY WLTNPFMILC GTVLYSTAII 151 LFQIILPHRP VQENVANAYE ALGSYLEAKA DFFDPDEAEW IGNRHIDLAM 201 SNTGVITAFN QCRSALFYRL RGKHRHPRTA KMLRYYFAAQ DIHERISSAH 251 VDYQEMSEKF KNTDIIFRIH RLLEMQGQAC RNTAQALRAS KDYVYSKRLG 301 RAIEGCRQSL RLLSDSNDNP DIRHLRRLLD NLGSVDQQFR QLQHNGLQAE 351 NDRMGDTRIA ALETGSLKNT WQAIRPQLNL ESGVFRHAVR LSLWAAACT 401 IVEALNLNLG YWILLTALFV CQPNYTATKS RVRQRIAGTV LGVIVGSLVP 451 YFTPSVETKL WIVIASTTLF FMTRTYKYSF STFFITIQAL TSLSLAGLDV 501 YAAMPVRIID TIIGASLAWA AVSYLWPDWK YLTLERTAAL AVCSNGAYLE 551 KITERLKSGE TGDDVEYRAT RRRAHEHTAA LSSTLSDMSS EPAKFADSLQ 601 PGFTLLKTGY ALTGYISALG AYRSEMHEEC SPDFTAQFHL AAEHTAHIFQ 651 HLPETEPDDF QTALDTLRGE LDTLRTHSSG TQSHILLQQL QLIARQLEPY 701 YRAYRQIPHR QPQNAA* ORF19a and ORF19-1 show 98.3% identity in 716 aa overlap: 10 20 30 40 50 60 orf19a. pep " MKTPPLKPLLITSLPVFASVFTAASIVWQLGEPKLAMPFVLGIIAGGLVDLDNRLTGRLK II II I II II II II I I I I I I I II I II II II II II I II I I I I I I I II I II II I II I I I I I I orf19-1 MKTPLLKPLLITSLPVFASVFTAASIVWQLGEPKLAMPFVLGIIAGGLVDLDNRLTGRLK 10 20 30 40 50 60 70 80 90 100 110 120 orf19a . pep NIIATVALFTLSSLVAQSTLGTGLPFILAMTLMTFGFTIMGAVGLKYRTFAFGALAVATY II I : I II I I I I I I I : I II I II II I I I I I I II II I I I I I I : II I II II I I II II I I I I I II orf19-1 NIITTVALFTLSSLTAQSTLGTGLPFILAMTLMTFGFTILGAVGLKYRTFAFGALAVATY 70 80 90 100 110 120 130 140 150 160 170 180 orf19a.pep TTLTYTPETYWLTNPFMILCGTVLYSTAIILFQIILPHRPVQENVANAYEALGSYLEAKA II I II II II II II II II II I II II I II I M II I M II II II I M II I II : II I: I I II I I orf19-1 TTLTYTPETYWLTNPFMILCGTVLYSTAILLFQIVLPHRPVQESVANAYDALGGYLEAKA 130 140 150 160 170 180 190 200 210 220 230 240 orf19a . pep DFFDPDEAEWIGNRHIDLAMSNTGVITAFNQCRSALFYRLRGKHRHPRTAKMLRYYFAAQ II I II II I I I II II II I I I I I II II II II II I II I II I II I II I I II I I I I I I II I I II orf19-1 DFFDPDEAAWIGNRHIDLAMSNTGVITAFNQCRSALFYRLRGKHRHPRTAKMLRYYFAAQ 190 200 210 220 230 240 250 260 270 280 290 300 orf19a.pep DIHERISSAHVDYQEMSEKFKNTDIIFRIHRLLEMQGQACRNTAQALRASKDYVYSKRLG II I II II II II II II II I I I I II I I I II II I I I I II II II I I II I I II I I I II I I I I I II orf19-1 DIHERISSAHVDYQEMSEKFKNTDIIFRIHRLLEMQGQACRNTAQALRASKDYVYSKRLG 250 260 270 280 290 300 310 320 330 340 350 360 orf19a. pep RAIEGCRQSLRLLSDSNDNPDIRHLRRLLDNLGSVDQQFRQLQHNGLQAENDRMGDTRIA I M I I I I I I II I I I II I M I I I II I I I I II II I I I I II I II II I I I I I I I I I I I II I II I orf19-1 RAIEGCRQSLRLLSDSNDSPDIRHLRRLLDNLGSVDQQFRQLQHNGLQAENDRMGDTRIA 310 320 330 340 350 360 370 380 390 400 410 420 orf19a. pep ALETGSLKNTWQAIRPQLNLESGVFRHAVRLSLWAAACTIVEALNLNLGYWILLTALFV II I I : II I I I I I II I I I II I II II I II I II II I I I I II I II I II I I II I II II I I I I I II orf19-1 ALETSSLKNTWQAIRPQLNLESGVFRHAVRLSLWAAACTIVEALNLNLGYWILLTALFV 370 380 390 400 410 420 430 440 450 460 470 480 orf19a .pep CQPNYTATKSRVRQRIAGTVLGVIVGSLVPYFTPSVETKLWIVIASTTLFFMTRTYKYSF I I I II I II II II I I I II I II I II I I II II II II I I I I I I I II I I I I I I II II I II I II I I orf19-1 CQPNYTATKSRVRQRIAGTVLGVIVGSLVPYFTPSVETKLWIVIASTTLFFMTRTYKYSF 430 440 450 460 470 480 490 500 510 520 530 540 orf19a. pep STFFITIQALTSLSLAGLDVYAAMPVRIIDTIIGASLAWAAVSYLWPDWKYLTLERTAAL I I II II I I I M I M M M I M I II I I I I II I I II I I II I I I M I II I II II II I I I I I I I orfl9-l STFFITIQALTSLSLAGLDVYAAMPVRIIDTIIGASLAWAAVSYLWPDWKYLTLERTAAL 490 500 510 520 530 540 550 560 570 580 590 600 orf19a. pep AVCSNGAYLEKITERLKSGETGDDVEYRATRRRAHEHTAALSSTLSDMSSEPAKFADSLQ I II I I I II II II I I I II II I II I I I I II II I II II I I I I I II I II I I I I II II I I I I I II orf19-1 AVCSNGAYLEKITERLKSGETGDDVEYRATRRRAHEHTAALSSTLSDMSSEPAKFADSLQ 550 560 570 580 590 600 610 620 630 640 650 660 orf19a. pep PGFTLLKTGYALTGYISALGAYRSEMHEECSPDFTAQFHLAAEHTAHIFQHLPETEPDDF I I I I II I I II II I II II I II I II II I II I I II II I I I I I I I I I I II I I I I II II I I II I I orf19-1 PGFTLLKTGYALTGYISALGAYRSEMHEECSPDFTAQFHLAAEHTAHIFQHLPETEPDDF 610 620 630 640 650 660 670 680 690 700 710 orf19a. pep QTALDTLRGELDTLRTHSSGTQSHILLQQLQLIARQLEPYYRAYRQIPHRQPQNAAX I II I I II I I I I I II II II I I II II I I I I I II I I I I I I II I II II I II II I I I II II I orf19-1 QTALDTLRGELDTLRTHSSGTQSHILLQQLQLIARQLEPYYRAYRQIPHRQPQNAAX 670 680 690 700 710 Homology with a predicted ORF from N.eonorrhoeae ORF 19 shows 95.1% identity over a 102aa overlap with a predicted ORF (ORF19.ng) from N gonorrhoeae: orf19. pep MKTPLLKPLLITSLPVFASVFTAASIVWQLGEPKLAMPFVLGIIAGGLVDLDΝXXTGRLK 60 II I I II I I I II I I II II I I I II I I II I II II I I II I I II II I II II I II I I I I II II I orfl9ng MKTPLLKPLLITSLPVFASVFTAASIVWQLGEPKLAMPFVLGIIAGGLVDLDΝRLTGRLK 60 orf19. pep ΝIITTVALFTLSSLTAQSTLGTGLPFILAMTLMTXXFTILGAX 103 II I : II I I I II II I I II II I I II I II I II II I I I I I I I II orfl9ng ΝIIATVALFTLSSLTAQSTLGTGLPFILAMTLMTFGFTILGAVGLKYRTFAFGALAVATY 120 An ORF19ng nucleotide sequence <SEQ ID 109> is predicted to encode a protein having amino acid sequence <SEQ ID 110>: 1 MKTPLLKPLL ITSLPVFASV FTAASIVWQL GEPKLAMPFV LGIIAGGLVD 51 LDΝRLTGRLK ΝIIATVALFT LSSLTAQSTL GTGLPFILAM TLMTFGFTIL 101 GAVGLKYRTF AFGALAVATY TTLTYTPETY WLTΝPFMILC GTVLYSTAII 151 LFQIILPHRP VQESVAΝAYE ALGGYLEAKA DFFDPDEAAW IGΝRHIDLAM 201 SΝTGVITAFΝ QCRSALFYRL RGKHRHPRTA KMLRYYFAAQ DIHERISSAH 251 VDYQEMSEKF KΝTDIIFRIR RLLEMQGQAC RΝTAQAIRSG KDYVYSKRLG 301 RAIEGCRQSL RLLSDGΝDSP DIRHLSRLLD ΝLGSVDQQFR QLRHSDSPAE 351 ΝDRMGDTRIA ALETGSFKΝT * Further work revealed the complete nucleotide sequence <SEQ ID 111>: 1 ATGAAAACCC CACTCCTCAA GCCTCTGCTC ATTACCTCGC TTCCCGTTTT 51 CGCCAGTGTC TTTACCGCCG CCTCCATCGT CTGGCAGCTA GGCGAACCCA 101 AGCTCGCCAT GCCCTTCGTA CTCGGCATCA TCGCCGGCGG CCTGGTCGAT 151 TTGGACAACC GCCTGACCGG ACGGCTGAAA AACATCATCG CCACCGTCGC 201 CCTGTTTACC CTCTCCTCGC TCACGGCGCA AAGCACCCTC GGCACAGGGC 251 TGCCCTTCAT CCTCGCCATG ACCCTGATGA CCTTCGGCTT TACCATTTTA 301 GGCGCGGTCG GGCTGAAATA CCGCACCTTC GCCTTCGGCG CACTCGCCGT 351 CGCCACCTAC ACCACGCTTA CCTACACCCC CGAAACCTAC TGGCTGACCA 401 ACCCCTTCAT GATTTTATGC GGCACCGTAC TGTACAGCAC CGCCATCATC 451 CTGTTCCAAA TCATCCTGCC CCACCGCCCC GTCCAAGAAA GCGTCGCCAA 501 TGCCTACGAA GCACTCGGCG GCTACCTCGA AGCCAAAGCC GACTTCTTCG 551 ACCCCGATGA GGCAGCCTGG ATAGGCAACC GCCACATCGA CCTCGCCATG 601 AGCAACACCG GCGTCATCAC CGCCTTCAAC CAATGCCGTT CCGCCCTGTT 651 TTACCGTTTG CGCGGCAAAC ACCGCCACCC GCGCACCGCC AAAATGCTGC 701 GCTACTACTT CGCCGCCCAA GACATCCACG AACGCATCAG CTCCGCCCAC 751 GTCGACTACC AAGAGATGTC CGAAAAATTC AAAAACACCG ACATCATCTT 801 CCGCATCCGC CGCCTGCTCG AAATGCAGGG GCAGGCGTGC CGCAACACCG 851 CCCAAGCCAT CCGGTCGGGC AAAGACTAcg tTTACAGCAA ACGCCTCGGA 901 CGCGCCATcg aaggctgCCG CCAGTCGCtg cgcctCCTTt cagacggcaA 951 CGACAGTCCC GACATCCGCC ACCTGAGccg CCTTCTCGAC AACCTCGgca 1001 GCGTcgacca gcagtTCcgc caactCCGAC ACAgcgactC CCCCGCcgaa 1051 Aacgaccgca tgggcgacaC CCGCATCGCC GCCCtcgaaa ccggcagctT 1101 caaaaaCAcc tggcaggCAA TCCGTCCGCa gctgaaCCTC GAATCatgCG 1151 TATTCCGCCA TGCCGTCCGC CTGTCCCTCG TCGTTGCCGC CGCCTGCACC 1201 ATCGTCgaag cCCTCAACCT CAACCTCGGC TACTGGATAC TGCTGACCGC 1251 CCTTTTCGTC TGCCAACCCA ACTACACCGC CACCAAAAGC CGCGTGTACC 1301 AACGCATCGC CGGCACCGTA CTCGGCGTAA TCGTCGGCTC GCTCGTCCCC 1351 TACTTCACCC CCTCCGTCGA AACCAAACTC TGGATTGTCA TCGCCGGTAC 1401 CACCCTGTTC TTCATGACCC GCACCTACAA ATACAGTTTC TCCACCTTCT 1451 TCATCACCAT TCAGGCACTG ACCAGCCTCT CCCTCGCAGG TTTGGACGTA 1501 TACGCCGCCA TGCCCGTGCG CATCATcgaC ACCATTATCG GCGCATCCCT 1551 TGCCTGGGCG GCGGTCAGCT ACCTGTGGCC AGACTGGAAA TACCTCACGC 1601 TCGAACGCAC CGCCGCCCTT GCCGTATGCA GCAGCGGCAC ATACCTCCAA 1651 AAAATTGCCG AACGCCTCAA AACCGGCGAA ACCGGCGACG ACATAGAATA 1701 CCGCATCACC CGCCGCCGCG CCCACGAACA CACCGCCGCC CTCAGCAGCA 1751 CCCTTTCCGA CATGAGCAGC GAACCCGCAA AATTCGCCGA CAGCCTGCAA 1801 CCCGGCTTTA CCCTGCTCAA AACCGGCTAC GCCCTGACCG GCTACATCTC 1851 CGCCCTCGGC GCATACCGCA GCGAAATGCA CGAAGAATGC AGCCCCGACT 1901 TTACCGCACA GTTCCACCTT GCCGCCGAAC ACACCGCCCA CATCTTCCAA 1951 CACCTGCCCG ACATGGGACC CGACGACTTT CAGACGGCAT TGGATACACT 2001 GCGCGGCGAA CTCGGCACCC TCCGCACCCG CAGCAGCGGA ACACAAAGCC 2051 ACATCCTCCT CCAACAGCTC CAACTCATCG CccgGCAACT CGAACCCTAC 2101 TACCGCGCCT ACCGACAAAT TCCGCACAGG CAGCCCCAAA ACGCAGCCTG 2151 A This corresponds to the amino acid sequence <SEQ ID 112; ORF19ng-l>: 1 MKTPLLKPLL ITSLPVFASV FTAASIVWQL GEPKLAMPFV LGIIAGGLVD 51 LDNRLTGRLK NIIATVALFT LSSLTAQSTL GTGLPFILAM TLMTFGFTIL 101 GAVGLKYRTF AFGALAVATY TTLTYTPETY WLTNPFMILC GTVLYSTAII 151 LFQIILPHRP VQESVANAYE ALGGYLEAKA DFFDPDEAAW IGNRHIDLAM 201 SNTGVITAFN QCRSALFYRL RGKHRHPRTA KMLRYYFAAQ DIHERISSAH 251 VDYQEMSEKF KNTDIIFRIR RLLEMQGQAC RNTAQAIRSG KDYVYSKRLG 301 RAIEGCRQSL RLLSDGNDSP DIRHLSRLLD NLGSVDQQFR QLRHSDSPAE 351 NDRMGDTRIA ALETGSFKNT WQAIRPQLNL ESCVFRHAVR LSLWAAACT 401 IVEALNLNLG YWILLTALFV CQPNYTATKS RVYQRIAGTV LGVIVGSLVP 451 YFTPSVETKL WIVIAGTTLF FMTRTYKYSF STFFITIQAL TSLSLAGLDV 501 YAAMPVRIID TIIGASLAWA AVSYLWPDWK YLTLERTAAL AVCSSGTYLQ 551 KIAERLKTGE TGDDIEYRIT RRRAHEHTAA LSSTLSDMSS EPAKFADSLQ 601 PGFTLLKTGY ALTGYISALG AYRSEMHEEC SPDFTAQFHL AAEHTAHIFQ 651 HLPDMGPDDF QTALDTLRGE LGTLRTRSΞG TQSHILLQQL QLIARQLEPY 701 YRAYRQIPHR QPQNAA* ORF19ng-l and ORF19-1 show 95.5% identity in 716 aa overlap: 10 20 30 40 50 60 orf19-1. pep MKTPLLKPLLITSLPVFASVFTAASIVWQLGEPKLAMPFVLGIIAGGLVDLDNRLTGRLK II II I I II I II II I I I II I II II II I I I I I I I I I I II II I II II II I I I I II I I I I II I I orfl9ng-l MKTPLLKPLLITSLPVFASVFTAASIVWQLGEPKLAMPFVLGIIAGGLVDLDNRLTGRLK 10 20 30 40 50 60 70 80 90 100 110 120 orf19-1. pep NIITTVALFTLSSLTAQSTLGTGLPFILAMTLMTFGFTILGAVGLKYRTFAFGALAVATY M I : II I I I I I II I II II I I I II I I II II I II I I II II I I I II II I I I II I I I I II I I II orfl9ng-l NIIATVALFTLSSLTAQSTLGTGLPFILAMTLMTFGFTILGAVGLKYRTFAFGALAVATY 70 80 90 100 110 120 130 140 150 160 170 180 orf19-1.pep TTLTYTPETYWLTNPFMILCGTVLYSTAILLFQIVLPHRPVQESVANAYDALGGYLEAKA I II I I II II I II I II II I II II I II I II I : I I I I : II II II I II I II I I : I I II II I II I orfl9ng-l TTLTYTPETYWLTNPFMILCGTVLYSTAIILFQIILPHRPVQESVANAYEALGGYLEAKA 130 140 150 160 170 180 190 200 210 220 230 240 orf19-1. pep DFFDPDEAAWIGNRHIDLAMSNTGVITAFNQCRSALFYRLRGKHRHPRTAKMLRYYFAAQ I II I I II II I I II II I II I I II I I II I II I II I II I II I II I I II I I I II I I I I II II II orfl9ng-l DFFDPDEAAWIGNRHIDLAMSNTGVITAFNQCRSALFYRLRGKHRHPRTAKMLRYYFAAQ 190 200 210 220 230 240 250 260 270 280 290 300 orf19-1. pep DIHERISSAHVDYQEMSEKFKNTDIIFRIHRLLEMQGQACRNTAQALRASKDYVYSKRLG II I II II I II I II II II II II II I I I II I : I I II II I II II I II II: I :: I II I II II I I orfl9ng-l DIHERISSAHVDYQEMSEKFKNTDIIFRIRRLLEMQGQACRNTAQAIRSGKDYVYSKRLG 250 260 270 280 290 300 310 320 330 340 350 360 orf19-1. pep RAIEGCRQSLRLLSDSNDSPDIRHLRRLLDNLGSVDQQFRQLQHNGLQAENDRMGDTRIA I I II I I I I I II II II : I II I I II II I I I I II II II II I I I I : I : I I II I I I I I I II orfl9ng-l RAIEGCRQSLRLLSDGNDSPDIRHLSRLLDNLGSVDQQFRQLRHSDSPAENDRMGDTRIA 310 320 330 340 350 360 370 380 390 400 410 420 orf19-1. pep ALETSSLKNTWQAIRPQLNLESGVFRHAVRLSLWAAACTIVEALNLNLGYWILLTALFV I II I : I: I I II I I I II I II I I I II I I I I I II II II II II I I I I I II I I II I I I I I I I I I orfl9ng-l ALETGSFKNTWQAIRPQLNLESCVFRHAVRLSLWAAACTIVEALNLNLGYWILLTALFV 370 380 390 400 410 420 430 440 450 460 470 480 orf19-1. pep CQPNYTATKSRVRQRIAGTVLGVIVGSLVPYFTPSVETKLWIVIASTTLFFMTRTYKYSF I I II I I I II I I I II I I I II I I II I I II II I I I I II I II I I I I II : I I I I II I I I I II I I orfl9ng-l CQPNYTATKSRVYQRIAGTVLGVIVGSLVPYFTPSVETKLWIVIAGTTLFFMTRTYKYSF 430 440 450 460 470 480 490 500 510 520 530 540 orf19-1. pep STFFITIQALTSLSLAGLDVYAAMPVRIIDTIIGASLAWAAVSYLWPDWKYLTLERTAAL I II I II I II I I II I II II II II I II I I II I II II II II II II II I II II I I I II I I I I II orfl9ng-l STFFITIQALTSLSLAGLDVYAAMPVRIIDTIIGASLAWAAVSYLWPDWKYLTLERTAAL 490 500 510 520 530 540 550 560 570 580 590 600 orf19-1.pep AVCSNGAYLEKITERLKSGETGDDVEYRATRRRAHEHTAALSSTLSDMSSEPAKFADSLQ II I I : I : I I : I I : I I II : I I II I M I II I I I II II II I I I I I I I II I I II II II I I II I orfl9ng-l AVCSSGTYLQKIAERLKTGETGDDIEYRITRRRAHEHTAALSSTLSDMSSEPAKFADSLQ 550 560 570 580 590 600 610 620 630 640 650 660 orf19-1. pep PGFTLLKTGYALTGYISALGAYRSEMHEECSPDFTAQFHLAAEHTAHIFQHLPETEPDDF I I II II I I II I I I I I II I I I I I II I I II II II II II II II I I I II II I II I I I : I I I I orfl9ng-l PGFTLLKTGYALTGYISALGAYRSEMHEECSPDFTAQFHLAAEHTAHIFQHLPDMGPDDF 610 620 630 640 650 660 670 680 690 700 710 orf19-1. pep QTALDTLRGELDTLRTHSSGTQSHILLQQLQLIARQLEPYYRAYRQIPHRQPQNAAX II I II II I I II I II I : I II I II I II I I I II II II II I I II I II II I II I II I I I I I orfl9ng-l QTALDTLRGELGTLRTRSSGTQSHILLQQLQLIARQLEPYYRAYRQIPHRQPQNAAX 670 680 690 700 710 In addition, ORF19ng-l shows significant homology to a hypothetical gonococcal protein previously entered in the databases: sp|033369|YOR2_NEIGO HYPOTHETICAL 45.5 KD PROTEIN (ORF2) gnl | PID | ell54438 (AJ002423) hypothetical protein [Neisseria gonorrh] Length = 417 Score = 1512 (705.6 bits), Expect = 5.3e-203, P = 5.3e-203 Identities = 301/326 (92%), Positives = 306/326 (93%) Query: 307 RQSLRLLSDGNDSPDIRHLSRLLDNLGSVDQQFRQLRHSDSPAENDRMGDTRIAALETGS 366 RQSLRLLSDGNDS DIRHLSRLLDNLGSVDQQFRQLRHSDSPAENDRMGDTRIAALETGΞ Sbjct: 1 RQSLRLLSDGNDSXDIRHLSRLLDNLGSVDQQFRQLRHSDSPAENDRMGDTRIAALETGS 60 Query: 367 FKNTWQAIRPQLNLESCVFRHAVRLSLWAAACTIVEALNLNLGYWILLTALFVCQPNYT 426 FKNTWQAIRPQLNLES VFRHAVRLSLWAAACTIVEALNLNLGYWILLT LFVCQPNYT Sbjct: 61 FKNTWQAIRPQLNLESGVFRHAVRLSLWAAACTIVEALNLNLGYWILLTRLFVCQPNYT 120 Query: 427 ATKSRVYQRIAGTVLGVIVGSLVPYFTPSVETKLWIVIAGTTLFFMTRTYKYSFSTFFIT 486 ATKSRVYQRIAGTVLGVIVGSLVPYFTPSVETKLWIVIAGTTLFFMTRTYKYSFSTFFIT Sbjct: 121 ATKSRVYQRIAGTVLGVIVGSLVPYFTPSVETKLWIVIAGTTLFFMTRTYKYSFSTFFIT 180 Query: 487 IQALTSLSLAGLDVYAAMPVRIIDTIIGASLAWAAVSYLWPDWKYLTLERTAALAVCSSG 546 IQALTSLSLAGLDVYAAMPVRIIDTIIGASLAWAAVSYLWPDWKYLTLERTAALAVCSSG Sbjct: 181 IQALTSLSLAGLDVYAAMPVRIIDTIIGASLAWAAVSYLWPDWKYLTLERTAALAVCSSG 240 Query: 547 TYLQKIAERLKTGETGDDIEYRITRRRAHEHTAALSSTLSDMSSEPAKFADSLQPGFTLL 606 TYLQKIAERLKTGETGDDIEYRITRRRAHEHTAALSSTLΞDMSSEPAKFAD+ P Sbjct: 241 TYLQKIAERLKTGETGDDIEYRITRRRAHEHTAALSSTLSDMSSEPAKFADTCNPALPCS 300 Query: 607 KTGYALTGYISALGAYRSEMHEECSP 632 K ALTGYISALG ++ + +P Sbjct: 301 KPATALTGYISALGHTAAKCTKNAAP 326 Based on this analysis, including the presence of several putative transmembrane domains in the gonococcal protein (the first of which is also seen in the meningococcal protein), and on homology with the YHFK protein, it is predicted that the proteins from N. meningitidis and N.gonorrhoeae, and their epitopes, could be useful antigens for vaccines or diagnostics, or for raising antibodies. Example 14 The following DNA sequence, believed to be complete, was identified in N. meningitidis <SEQ D 113>: 1 ATGAATATGC TGGGAGCTTT GGCAAAAGTC GGCAGCCTGA CGATGGTGTC 51 GCGCGTTTTG GGATTTGTGC GCGATACGGT CATTGCGCGG GCATTCGGCG 101 CGGGTATGGC GACGGATGCG TTTTTTGTCG CGTTCAAACT GCCCAACCTG 151 CTTCGCCGCG TGTTTGCGGA GGGGGCGTTT GCCCAAGCGT TTGTGCCGAT 201 TTTGGCGGAA TACAAGGAAA CGCGTTCAAA AGAGGCGG . C GAAGCCTTTA 251 TCCGCCATGT GGCGGGGATG CTGTCGTTTG TACTGGTTAT CGTTACCGCG 301 CTGGGCATAC TTGCCGCGCC TTGGGTGATT TATGTTTCCG CACCCGAGTT 351 TTGCCCAAGA TGCCGACAAA TTTCAGCTCT CCATCGATTT GCTGCGGATT 401 ACGTTTCCTT ATATATTATT GATTTCCCTG TCTTCATTTG TCGGCTCGGT 451 ACTCAATTCT TATCATAAGT TCGGCATTCC GGCGTTTACG CCAC . GTTTC 501 TGAACGTGTC GTTTATCGTA TTCGCGCTGT TTTTCGTGCC GTATTTCGAT 551 CCGCCCGTTA CCGCGCyGGC GTGGGCGGTC TTTGTCGGCG GCATTTTGCA 601 ACTCGrmTTC CAACTGCCCT GGCTGGCGAA ACTGGGCTTT TTGAAACTGC 651 CCAAACtGAG TTTCAAAGAT GCGGCGGTCA ACCGCGTGAT GAAACAGATG 701 GCGCCTGCgA TTTTgGGCGT GAgCGTGGCG CAGGTTTCTT TGGTGATCAA 751 CACGATTTTc GCGTCTTATC TGCAATCGGG CAGCGTTTCA TGGATGTATT 801 ACGCCGACCG CATGATGGAG CTGCCCAGCG GCGTGCTGGG GGCGGCACTC 851 GGTACGATTT TGCTGCCGAC TTTGTCCAAA CACTCGGCAA ACCaAGATAC 901 GGaACAGTTT TCCGCCCTGC TCGACTGGGG TTTGCGCCTG TGCATGCtgc 951 TGACGCTGCC GGCGgcGGTC GGACTGGCGG TGTTGTCGTT cCCgCtGGTG 1001 GCGACGCTGT TTATGTACCG CGwATTTACG CTGTTTGACG CGCAGATGAC 1051 GCAACACGCG CTGATTGCCT ATTCTTTCGG TTTAATCGGC TTAATCATGA 1101 TTAAAGTGTT GGCACCCGGC TTCTATGCGC GGCAAAACAT CAA AmGCCC 1151 GTCAAAATCG CCATCTTCAC GCTCATCTGC mCGCAGTTGA TGAACCTTGs 1201 CTTTAyCGGC CCACTrrAAC rCasTCGGAC TTTCGCTTGC CATCGGTCTG 1251 GGCGCGTGTA TCAATGCCGG ATTGTTGTTT TACCTGTTGC GCAGACACGG 1301 TATTTACCAA CCTGG . CAAG GGTTGGGCAG CGTTCTT . AG CAAAAATGCT 1351 GcTCTCGCTC GCCGTGA This corresponds to the amino acid sequence <SEQ ID 114; ORF20>: 1 MNMLGALAKV GSLTMVSRVL GFVRDTVIAR AFGAGMATDA FFVAFKLPNL 51 LRRVFAEGAF AQAFVPILAE YKETRSKEAX EAFIRHVAGM LSFVLVIVTA 101 LGILAAPWVI YVSAPSFAQD ADKFQLSIDL LRITFPYILL ISLSSFVGSV 151 LNSYHKFGIP AFTPXFLNVS FIVFALFFVP YFDPPVTAXA WAVFVGGILQ 201 LXFQLPWLAK LGFLKLPKLS FKDAAVNRVM KQMAPAILGV SVAQVSLVIN 251 TIFASYLQSG SVSWMYYADR MMELPSGVLG AALGTILLPT LSKHSANQDT 301 EQFSALLDWG LRLCMLLTLP AAVGLAVLSF PLVATLFMYR XFTLFDAQMT 351 QHALIAYSFG LIGLIMIKVL APGFYARQNI XXPVKIAIFT LICXQLMNLX 401 FXGPLXXIGL SLAIGLGACI NAGLLFYLLR RHGIYQPXQG LGSVLXQKCC 451 SRSP* These sequences were elaborated, and the complete DNA sequence <SEQ ID 115> is: 1 ATGAATATGC TGGGAGCTTT GGCAAAAGTC GGCAGCCTGA CGATGGTGTC 51 GCGCGTTTTG GGATTTGTGC GCGATACGGT CATTGCGCGG GCATTCGGCG 101 CGGGTATGGC GACGGATGCG TTTTTTGTCG CGTTCAAACT GCCCAACCTG 151 CTTCGCCGCG TGTTTGCGGA GGGGGCGTTT GCCCAAGCGT TTGTGCCGAT 201 TTTGGCGGAA TACAAGGAAA CGCGTTCAAA AGAGGCGGCG GAGGCTTTTA 251 TCCGCCATGT GGCGGGGATG CTGTCGTTTG TACTGGTTAT CGTTACCGCG 301 CTGGGCATAC TTGCCGCGCC TTGGGTGATT TATGTTTCCG CACCCGGTTT 351 TGCCCAAGAT GCCGACAAAT TTCAGCTCTC CATCGATTTG CTGCGGATTA 401 CGTTTCCTTA TATATTATTG ATTTCCCTGT CTTCATTTGT CGGCTCGGTA 451 CTCAATTCTT ATCATAAGTT CGGCATTCCG GCGTTTACGC CCACGTTTCT 501 GAACGTGTCG TTTATCGTAT TCGCGCTGTT TTTCGTGCCG TATTTCGATC 551 CGCCCGTTAC CGCGCTGGCG TGGGCGGTCT TTGTCGGCGG CATTTTGCAA 601 CTCGGCTTCC AACTGCCCTG GCTGGCGAAA CTGGGCTTTT TGAAACTGCC 651 CAAACTGAGT TTCAAAGATG CGGCGGTCAA CCGCGTGATG AAACAGATGG 701 CGCCTGCGAT TTTGGGCGTG AGCGTGGCGC AGGTTTCTTT GGTGATCAAC 751 ACGATTTTCG CGTCTTATCT GCAATCGGGC AGCGTTTCAT GGATGTATTA 801 CGCCGACCGC ATGATGGAGC TGCCCAGCGG CGTGCTGGGG GCGGCACTCG 851 GTACGATTTT GCTGCCGACT TTGTCCAAAC ACTCGGCAAA CCAAGATACG 901 GAACAGTTTT CCGCCCTGCT CGACTGGGGT TTGCGCCTGT GCATGCTGCT 951 GACGCTGCCG GCGGCGGTCG GACTGGCGGT GTTGTCGTTC CCGCTGGTGG 1001 CGACGCTGTT TATGTACCGC GAATTTACGC TGTTTGACGC GCAGATGACG 1051 CAACACGCGC TGATTGCCTA TTCTTTCGGT TTAATCGGCT TAATCATGAT 1101 TAAAGTGTTG GCACCCGGCT TCTATGCGCG GCAAAACATC AAAACGCCCG 1151 TCAAAATCGC CATCTTCACG CTCATCTGCA CGCAGTTGAT GAACCTTGCC 1201 TTTATCGGCC CACTGAAACA CGTCGGACTT TCGCTTGCCA TCGGTCTGGG 1251 CGCGTGTATC AATGCCGGAT TGTTGTTTTA CCTGTTGCGC AGACACGGTA 1301 TTTACCAACC TGGCAAGGGT TGGGCAGCGT TCTTAGCAAA AATGCTGCTC 1351 TCGCTCGCCG TGATGTGCGG CGGACTGTGG GCAGCGCAGG CTTACCTGCC 1401 GTTTGAATGG GCGCACGCCG GCGGAATGCG GAAAGCGGGG CAGCTCTGCA 1451 TCCTGATTGC CGTCGGCGGC GGACTGTATT TCGCATCACT GGCGGCTTTG 1501 GGCTTCCGTC CGCGCCATTT CAAACGCGTG GAAAACTGA This coπesponds to the amino acid sequence <SEQ ID 116; ORF20-1>: 1 MNMLGALAKV GSLTMVSRVL GFVRDTVIAR AFGAGMATDA FFVAFKLPNL 51 LRRVFAEGAF AQAFVPILAE YKETRSKEAA EAFIRHVAGM LSFVLVIVTA 101 LGILAAPWVI YVSAPGFAQD ADKFQLSIDL LRITFPYILL ISLSSFVGSV 151 LNSYHKFGIP AFTPTFLNVS FIVFALFFVP YFDPPVTALA WAVFVGGILQ 201 LGFQLPWLAK LGFLKLPKLS FKDAAVNRVM KQMAPAILGV SVAQVSLVIN 251 TIFASYLQSG SVSWMYYADR MMELPSGVLG AALGTILLPT LSKHSANQDT 301 EQFSALLDWG LRLCMLLTLP AAVGLAVLSF PLVATLFMYR EFTLFDAQMT 351 QHALIAYSFG LIGLIMIKVL APGFYARQNI KTPVKIAIFT LICTQLMNLA 401 FIGPLKHVGL SLAIGLGACI NAGLLFYLLR RHGIYQPGKG WAAFLAKMLL 451 SLAVMCGGLW AAQAYLPFEW AHAGGMRKAG QLCILIAVGG GLYFASLAAL 501 GFRPRHFKRV EN* Computer analysis of this amino acid sequence gave the following results: Homology with the MviN virulence factor of S. typhimurium (accession number P37169) ORF20 and MviN proteins show 63% aa identity in 440aa overlap: Orf20 1 MNMLGALAKVGSLTMVSRVLGFVRDTVIARAFGAGMATDAFFVAFKLPNLLRRVFAEGAF 60 MN+L +LA V S+TM SRVLGF RD ++AR FGAGMATDAFFVAFKLPNLLRR+FAEGAF MviN 14 MNLLKSLAAVSSMTMFSRVLGFARDAIVARIFGAGMATDAFFVAFKLPNLLRRIFAEGAF 73 Orf20 61 AQAFVPILAEYKETRSKEAXEAFIRHVAGMLSFVLVIVTALGILAAPWVIYVSAPSFAQD 120 +QAFVPILAEYK + +EA F+ +V+G+L+ L +VT G+LAAPWVI V+AP FA MviN 74 SQAFVPILAEYKSKQGEEATRIFVAYVSGLLTLALAWTVAGMLAAPWVIMVTAPGFADT 133 Orf20 121 ADKFQLSIDLLRITFPYILLISLSSFVGSVLNSYHKFGIPAFTPXFLNVSFIVFALFFVP 180 ADKF L+ LLRITFPYILLISL+S VG++LN++++F IPAF P FLN+S I FALF P MviN 134 ADKFALTTQLLRITFPYILLISLASLVGAILNTWNRFSIPAFAPTFLNISMIGFALFAAP 193 Orf20 181 YFDPPVTAXAWAVFVGGILQLXFQLPWLAKLGFLKLPKLSFKDAAVNRVMKQMAPAILGV 240 YF+PPV A AWAV VGG+LQL +QLP+L K+G L LP+++F+D RV+KQM PAILGV MviN 194 YFNPPVLALAWAVTVGGVLQLVYQLPYLKKIGMLVLPRINFRDTGAMRWKQMGPAILGV 253 Orf20 241 SVAQVSLVINTIFASYLQSGSVSWMYYADRMMELPSGVLGAALGTILLPTLSKHSANQDT 300 SV+Q+SL+INTIFAS+L SGSVSWMYYADR+ME PSGVLG ALGTILLP+LSK A+ + MviN 254 SVSQISLIINTIFASFLASGSVSWMYYADRLMEFPSGVLGVALGTILLPSLSKSFASGNH 313 Orf20 301 EQFSALLDWGLRLCMLLTLPAAVGLAVLSFPLVATLFMYRXFTLFDAQMTQHALIAYSFG 360 +++ L+DWGLRLC LL LP+AV L +L+ PL +LF Y FT FDA MTQ ALIAYS G MviN 314 DEYCRLMDWGLRLCFLLALPSAVALGILAKPLTVSLFQYGKFTAFDAAMTQRALIAYSVG 373 Orf20 361 LIGLIMIKVLAPGFYARQNIXXPVKIAIFTLICXQLMNLXFXXXXXXXXXXXXXXXXXCI 420 LIGLI++KVLAPGFY+RQ+I PVKIAI TLI QLMNL F C+ MviN 374 LIGLIWKVLAPGFYSRQDIKTPVKIAIVTLIMTQLMNLAFIGPLKHAGLSLSIGLAACL 433 Orf20 421 NAGLLFYLLRRHGIYQPXQG 440 NA LL++ LR+ 1+ P G MviN 434 NASLLYWQLRKQNIFTPQPG 453 Homology with a predicted ORF from N.meninsitidis (strain Al ORF20 shows 93.5% identity over a 447aa overlap with an ORF (ORF20a) from strain A of N meningitidis: 10 20 30 40 50 60 orf20. pep MΝMLGALAKVGSLTMVSRVLGFVRDTVIARAFGAGMATDAFFVAFKLPΝLLRRVFAEGAF I I I II I I : I II I II I I I II I II I I I I I II II II II II II II I I I I II I II I I I II II II I orf20a MΝMLGALVKVGSLTMVSRVLGFVRDTVIARAFGAGMATDAFFVAFKLPΝLLRRVFAEGAF 10 20 30 40 50 60 70 80 90 100 110 120 orf20. pep AQAFVPILAEYKETRSKEAXEAFIRHVAGMLSFVLVIVTALGILAAPWVIYVSAPSFAQD I I II I II I I I I II II I I IM II I I II II I II II I I I I I I II II I II II I II II II: I I: I orf20a AQAFVPILAEYKETRSKEATEAFIRHVAGMLSFVLVIVTALGILAAPWVIYVSAPGFAKD 70 80 90 100 110 120 130 140 150 160 170 180 orf20.pep ADKFQLSIDLLRITFPYILLISLSSFVGSVLΝSYHKFGIPAFTPXFLΝVSFIVFALFFVP I II I I I II I I I II II I I I I I II I I I I I I I I II I I II M II I I I I : I II II II I II I I I I I orf20a ADKFQLSIDLLRITFPYILLISLSSFVGSVLΝSYHKFSIPAFTPTFLΝVSFIVFALFFVP 130 140 150 160 170 180 190 200 210 220 230 240 orf20. pep YFDPPVTAXAWAVFVGGILQLXFQLPWLAKLGFLKLPKLSFKDAAVΝRVMKQMAPAILGV I II I I II I I I I II I I II II I II I II I II I I II I I II II II I II II II I I I I II II I II orf20a YFDPPVTALAWAVFVGGILQLGFQLPWLAKLGFLKLPKLSFKDAAVΝRVMKQMAPAILGV 190 200 210 220 230 240 250 260 270 280 290 300 orf20. pep SVAQVSLVIΝTIFASYLQSGSVSWMYYADRMMELPSGVLGAALGTILLPTLSKHSAΝQDT I I I I : I II I I I I I I I II I I II I I I I I I II II II I I : I I I I I II I I I II I I II II I I I I I I orf20a SVAQISLVIΝTIFASYLQSGSVSWMYYADRMMELPGGVLGAALGTILLPTLSKHSAΝQDT 250 260 270 280 290 300 310 320 330 340 350 360 orf20. pep EQFSALLDWGLRLCMLLTLPAAVGLAVLSFPLVATLFMYRXFTLFDAQMTQHALIAYSFG II I I I II I II I I II I I I II I I I I : II I II I II II II II I I II I II II I I I II I I I I I I orf20a EQFSALLDWGLRXCMLLTLPAAVGMAVLSFPLVATLFMYREFTLFDAQMTQHALIAYSFG 310 320 330 340 350 360 370 380 390 400 410 420 orf20.pep LIGLIMIKVLAPGFYARQΝIXXPVKIAIFTLICXQLMΝLXFXGPLXXIGLSLAIGLGACI I I I II I II I II I II I II II I : I I II I II I II I : I I I I I I III : II I I I II II I II orf20a LIGLIMIKVLAPGFYARQΝIKTPVKIAIFTLICTQLMΝLAFIGPLKHVGLSLAIGLGACI 370 380 390 400 410 420 430 440 450 orf20.pep ΝAGLLFYLLRRHGIYQPXQGLGSVLXQKCCSRSPX I I I II I II I I II I I I I I : I :: I : orf20a ΝAGLLFYLLRRHGIYQPGKGWAAFLAKMLLSLAVMGGGLYAAQIWLPFDWAHAGGMQKAA 430 440 450 460 470 480 The complete length ORF20a nucleotide sequence <SEQ LD 117> is: 1 ATGAATATGC TGGGAGCTTT GGTAAAAGTC GGCAGCCTGA CGATGGTGTC 51 GCGCGTTTTG GGATTTGTGC GCGATACGGT CATTGCGCGC GCATTCGGCG 101 CAGGCATGGC GACGGATGCG TTCTTTGTCG CGTTCAAACT GCCCAACCTG 151 CTTCGCCGCG TGTTTGCGGA GGGGGCGTTT GCCCAAGCGT TTGTGCCGAT 201 TTTGGCGGAA TATAAGGAAA CGCGTTCTAA AGAGGCGACG GAGGCTTTTA 251 TCCGCCATGT GGCGGGGATG CTGTCGTTTG TACTGGTCAT CGTTACCGCG 301 CTGGGCATAC TTGCCGCGCC TTGGGTGATT TATGTTTCCG CACCCGGTTT 351 TGCCAAAGAT GCCGACAAAT TTCAGCTCTC TATCGATTTG CTGCGGATTA 401 CGTTTCCTTA TATCTTATTG ATTTCACTTT CCTCTTTTGT CGGCTCGGTA 451 CTCAATTCCT ATCATAAATT CAGCATTCCT GCGTTTACGC CCACGTTCCT 501 GAACGTGTCG TTTATCGTAT TCGCGCTGTT TTTCGTGCCG TATTTCGATC 551 CTCCCGTTAC CGCGCTGGCT TGGGCGGTTT TTGTCGGCGG CATTTTGCAA 601 CTCGGCTTCC AACTGCCCTG GCTGGCGAAA CTGGGTTTTT TGAAACTGCC 651 CAAACTGAGT TTCAAAGATG CGGCGGTCAA CCGCGTGATG AAACAGATGG 701 CGCCTGCGAT TTTGGGCGTG AGCGTGGCGC AGATTTCTTT GGTGATCAAC 751 ACGATTTTCG CGTCTTATCT GCAATCGGGC AGCGTTTCAT GGATGTATTA 801 CGCCGACCGC ATGATGGAAC TGCCCGGCGG CGTGCTGGGG GCGGCACTCG 851 GTACGATTTT GCTGCCGACT TTGTCCAAAC ACTCGGCAAA CCAAGATACG 901 GAACAGTTTT CCGCCCTGCT CGACTGGGGT TTGCGCNTGT GCATGCTGCT 951 GACGCTGCCG GCGGCGGTCG GAATGGCGGT GTTGTCGTTC CCGCTGGTGG 1001 CAACCTTGTT TATGTACCGA GAATTCACGC TGTTTGACGC GCAGATGACG 1051 CAACACGCGC TGATTGCCTA TTCTTTCGGT TTAATCGGTT TAATCATGAT 1101 TAAAGTGTTG GCGCCCGGCT TTTATGCGCG GCAAAACATC AAAACGCCCG 1151 TCAAAATCGC CATCTTCACG CTCATTTGCA CGCAGTTGAT GAACCTTGCC 1201 TTTATCGGCC CACTGAAACA CGTCGGACTT TCGCTTGCCA TCGGTCTGGG 1251 CGCGTGTATC AATGCCGGAT TGTTGTTTTA CCTGTTGCGC AGACACGGTA 1301 TTTACCAACC TGGCAAGGGT TGGGCAGCGT TCTTGGCAAA AATGCTGCTC 1351 TCGCTCGCCG TGATGGGAGG CGGCCTGTAT GCCGCCCAAA TCTGGCTGCC 1401 GTTCGACTGG GCACACGCCG GCGGAATGCA AAAGGCCGCC CGGCTCTTCA 1451 TCCTGATTGC CGTCGGCGGC GGACTGTATT TCGCATCACT GGCGGCTTTG 1501 GGCTTCCGTC CGCGCCATTT CAAACGCGTG GAAAGCTGA This encodes a protein having amino acid sequence <SEQ ID 118>: 1 MNMLGALVKV GΞLTMVSRVL GFVRDTVIAR AFGAGMATDA FFVAFKLPNL 51 LRRVFAEGAF AQAFVPILAE YKETRSKEAT EAFIRHVAGM LSFVLVIVTA 101 LGILAAPWVI YVSAPGFAKD ADKFQLSIDL LRITFPYILL ISLSSFVGSV 151 LNSYHKFSIP AFTPTFLNVS FIVFALFFVP YFDPPVTALA WAVFVGGILQ 201 LGFQLPWLAK LGFLKLPKLS FKDAAVNRVM KQMAPAILGV SVAQISLVIN 251 TIFASYLQSG SVSWMYYADR MMELPGGVLG AALGTILLPT LSKHSANQDT 301 EQFSALLDWG LRXCMLLTLP AAVGMAVLSF PLVATLFMYR EFTLFDAQMT 351 QHALIAYSFG LIGLIMIKVL APGFYARQNI KTPVKIAIFT LICTQLMNLA 401 FIGPLKHVGL SLAIGLGACI NAGLLFYLLR RHGIYQPGKG WAAFLAKMLL 451 SLAVMGGGLY AAQIWLPFDW AHAGGMQKAA RLFILIAVGG GLYFASLAAL 501 GFRPRHFKRV ES* ORF20a and ORF20-1 show 96.5% identity in 512 aa overlap: 10 20 30 40 50 60 orf20a . pep MNMLGALVKVGSLTMVSRVLGFVRDTVIARAFGAGMATDAFFVAFKLPNLLRRVFAEGAF II I I I I I : II II I II I I I I II I II II II II I I I II II I I I I I I I II I II I II I I I I I I II orf20-1 MNMLGALAKVGSLTMVSRVLGFVRDTVIARAFGAGMATDAFFVAFKLPNLLRRVFAEGAF 10 20 30 40 50 60 70 80 90 100 110 120 orf20a . pep AQAFVPILAEYKETRSKEATEAFIRHVAGMLSFVLVIVTALGILAAPWVIYVSAPGFAKD II I I I I I I II II I II I II M I II I I I I I I II I I I I I I I I I II II I I I I I I I I I II I I M I orf20-1 AQAFVPILAEYKETRSKEAAEAFIRHVAGMLSFVLVIVTALGILAAPWVIYVSAPGFAQD 70 80 90 100 110 120 130 140 150 160 170 180 orf20a. pep ADKFQLSIDLLRITFPYILLISLSSFVGSVLNSYHKFSIPAFTPTFLNVSFIVFALFFVP I I I II II II I II I II I I II I I II I I II II II I I I I I I : I I I II II I I I II I I I I I II I II orf20-1 ADKFQLSIDLLRITFPYILLISLSSFVGSVLNSYHKFGIPAFTPTFLNVSFIVFALFFVP 130 140 150 160 170 180 190 200 210 220 230 240 orf20a . pep YFDPPVTALAWAVFVGGILQLGFQLPWLAKLGFLKLPKLSFKDAAVNRVMKQMAPAILGV I I II II I II I II I II I II I I I II I I II II II I I I II II I II II II I II II I II I I I II I I orf20-1 YFDPPVTALAWAVFVGGILQLGFQLPWLAKLGFLKLPKLSFKDAAVNRVMKQMAPAILGV 190 200 210 220 230 240 250 260 270 280 290 300 orf20a. pep SVAQISLVINTIFASYLQSGSVSWMYYADRMMELPGGVLGAALGTILLPTLSKHSANQDT II I I : II II I II I I I I I I II II I I I II II I I II II : I II I I I I I I I I I I I I II I I I I II I orf20-1 SVAQVSLVINTIFASYLQSGSVSWMYYADRMMELPSGVLGAALGTILLPTLSKHSANQDT 250 260 270 280 290 300 310 320 330 340 350 360 orf20a . pep EQFSALLDWGLRXCMLLTLPAAVGMAVLSFPLVATLFMYREFTLFDAQMTQHALIAYSFG I II I I I I II I II I II I I II I I I I : II I I II I II II II I I I II I II I I I I II I I II I I II orf20-1 EQFSALLDWGLRLCMLLTLPAAVGLAVLSFPLVATLFMYREFTLFDAQMTQHALIAYSFG 310 320 330 340 350 360 370 380 390 400 410 420 orf20a. pep LIGLIMIKVLAPGFYARQNIKTPVKIAIFTLICTQLMNLAFIGPLKHVGLSLAIGLGACI I I I II I II I I II II I II I II I I I II I I I I II I II II I I II I II II I II I II II I I I I I I I orf20-1 LIGLIMIKVLAPGFYARQNIKTPVKIAIFTLICTQLMNLAFIGPLKHVGLSLAIGLGACI 370 380 390 400 410 420 430 440 450 460 470 480 orf20a . pep NAGLLFYLLRRHGIYQPGKGWAAFLAKMLLSLAVMGGGLYAAQIWLPFDWAHAGGMQKAA II I II I I I I I II II II I I I II II I I I II I I II II I I I I : II I : II M I I I II I M II : orf20-1 NAGLLFYLLRRHGIYQPGKGWAAFLAKMLLSLAVMCGGLWAAQAYLPFEWAHAGGMRKAG 430 440 450 460 470 480 490 500 510 orf20a . pep RLFILIAVGGGLYFASLAALGFRPRHFKRVESX : I I I II II I I I I II II I II I II I I I I I I I I : I orf20-1 QLCILIAVGGGLYFASLAALGFRPRHFKRVENX 490 500 510 Homology with a predicted ORF from N.gonorrhoeae ORF20 shows 92.1% identity over a 454aa overlap with a predicted ORF (ORF20ng) from N. gonorrhoeae: orf20.pep MNMLGALAKVGSLTMVSRVLGFVRDTVIARAFGAGMATDAFFVAFKLPNLLRRVFAEGAF 60 II II I II I I I II I II II II II I II I I II I I I I II I I I II I I I I I II I I I II II II I I I I I orf20ng MNMLGALAKVGSLTMVSRVLGFVRDTVIARAFGAGMATDAFFVAFKLPNLLRRVFAEGAF 60 orf20.pep AQAFVPILAEYKETRSKEAXEAFIRHVAGMLSFVLVIVTALGILAAPWVIYVSAPSFAQD 120 I I II I II I II II I II I I I I : I I II II I I I I I I II I :: I I I I I II II I I I II II II : I :: I orf20ng AQAFVPILAEYKETRSKEATEAFIRHVAGMLSFVLIWTALGILAAPWVIYVSAPGFTKD 120 orf20.pep ADKFQLSIDLLRITFPYILLISLSSFVGSVLNSYHKFGIPAFTPXFLNVSFIVFALFFVP 180 II I II I I I : II I I I II II I I I II I II I I I: I II I II II II I II I : II I : I II I I I II I I I orf20ng ADKFQLSISLLRITFPYILLISLSSFVGSILNSYHKFGIPAFTPTFLNISFIVFALFFVP 180 orf20.pep YFDPPVTAXAWAVFVGGILQLXFQLPWLAKLGFLKLPKLSFKDAAVNRVMKQMAPAILGV 240 I I I I I II I I II I II II II I I I II I I I I II I I II II I M I I I I I I II II II I I II I I II orf20ng YFDPPVTALAWAVFVGGILQLGFQLPWLAKLGFLKLPKLNFKDAAVNRVMKQMAPAILGV 240 orf20.pep SVAQVSLVINTIFASYLQSGSVSWMYYADRMMELPSGVLGAALGTILLPTLSKHSANQDT 300 I II I : I I I I I I I II I I I I I II I II I II I II II II I : II I I I I II II II II I II II I I I I I orf20ng SVAQISLVINTIFASYLQSGSVSWMYYADRMMELPGGVLGAALGTILLPTLSKHSANQDT 300 orf20.pep EQFSALLDWGLRLCMLLTLPAAVGLAVLSFPLVATLFMYRXFTLFDAQMTQHALIAYSFG 360 I I II II II II I II I II I I II I M I II I II I I II I II II II II II II I I II II II I I I I I orf20ng EQFSALLDWGLRLCMLLTLPAAAGLAVLSFPLVATLFMYREFTLFDAQMTQHALIAYSFG 360 orf20.pep LIGLIMIKVLAPGFYARQNIXXPVKIAIFTLICXQLMNLXFXGPLXXIGLSLAIGLGACI 420 II I II I I II I I II I I I I I I : I II II I II II I: I II II I III II II II I II II I orf20ng LIGLIMIKVLASGFYARQNIKTPVKIAIFTLICTQLMNLAFIGPLKHAGLSLAIGLGACI 420 orf20.pep NAGLLFYLLRRHGIYQPXQGLGSVLXQKCCSRS 454 II I I II : I : I : I II I : I IMM : II II II orf20ng NAGLLFFLFRKHGIYRPGQGLGQPSWRKCCSRSP 445544 An ORF20ng nucleotide sequence <SEQ LD 119> was predicted to encode a protein having amino acid sequence <SEQ ID 120> 1 MNMLGALAKV GSLTMVSRVL GFVRDTVIAR AFGAGMATDA FFVAFKLPNL 51 LRRVFAEGAF AQAFVPILAE YKETRSKEAT EAFIRHVAGM LSFVLIWTA 101 LGILAAPWVI YVSAPGFTKD ADKFQLSISL LRITFPYILL ISLSSFVGSI 151 LNSYHKFGIP AFTPTFLNIS FIVFALFFVP YFDPPVTALA WAVFVGGILQ 201 LGFQLPWLAK LGFLKLPKLN FKDAAVNRVM KQMAPAILGV SVAQISLVIN 251 TIFASYLQSG SVSWMYYADR MMELPGGVLG AALGTILLPT LSKHSANQDT 301 EQFSALLDWG LRLCMLLTLP AAAGLAVLSF PLVATLFMYR EFTLFDAQMT 351 QHALIAYSFG LIGLIMIKVL ASGFYARQNI KTPVKIAIFT LICTQLM LA 401 FIGPLKHAGL SLAIGLGACI NAGLLFFLFR KHGIYRPGQG LGQPSWRKCC 451 SRSP* Further DNA sequence analysis revealed the following DNA sequence <SEQ LD 121>: 1 ATGAATATGC TTGGAGCTTT GGCAAAAGTC GGCAGCCTGA CGATGGTGTC 51 GCGCGTTTTG GGATTTGTGC GCGATACGGT CATTGCGCGG GCATTCGGCG 101 CGGGTATGGC GACGGATGCG TTTTTTGTCG CGTTCAAACT GCCCAACCTG 151 CTTCGCCGCG TGTTTGCGGA GGGGGCGTTT GCCCAAGCGT TTGTGCCGAT 201 TTTGGCGGAA TATAAGGAAA CGCGTTCTAA AGAGGCGAcg gAGGCTTTTA 251 TCCGCCACGt tgcgggAatg CTGTCGTTTG TGCTGATcgt cGttacCGCG 301 CTGGGCATAC TTGCCGCgcc tTGGGTGATT TATGTTtccg CgcccGGCTT 351 TACCAAAGAC GCGGACAAGT TCCAACTTTC CATCAGCCTG CTGCGGATTA 401 CGTTTCCTTA TATATTATTG ATTTCTTTGT CTTCTTTTGT CGGCTCGATA 451 CTCAATTCCT ACCATAAGTT CGGCATTCCC GCGTTTACGC CCACGTTTTT 501 AAACATCTCT TTTATCGTAT TCGCACTGTT TTTCGTGCCG TATTTCGATC 551 CGCCCGTTAC CGCGCTGGCG TGGGCGGTTT TTGTCGGCGG TATTTTGCAG 601 CTCGGTTTCC AACTGCCGTG GCTGGCGAAA CTGGGCTTTT TGAAACTGCC 651 CAAACTGAAT TTCAAAGATG CGGCGGTCAA CCGCGTCATG AAACAGATGG 701 CGCCTGCGAT TTTGGGCGTG agcgTGGCGC AAATTTCTTT GgttATCAAC 751 ACGATTTTCG CGTCTTATCT GCAATCGGGC AGCGTTTCAT GGATGTatta 801 cgCCGACCGC ATGATGGAGc tgcgccGGGG CGTGCTGGGG GCTGCACTCG 851 GTACAATTTT GCTGCCGACT TTGTCCAAAC ACTCGGCAAA CCAAGATACG 901 GAACAGTTTT CCGCCCTGCT CGACTGGGGT TTGCGCCTGT GCATGCTGCT 951 GACGCTGCCG GCGGCGGccg GACTGGCGGT ATTGTCGTTC CCGCTGGTGG 1001 CGACGCTGTT TATGTACCGA GAATTCACGC TGTTTGACGC ACAAATGACG 1051 CAACACGCGC TGATTGCCTA TTCTTTCGGT TTAATCGGTT TAATTATGAT 1101 TAAAGTGTTG GCATCCGGCT TTTATGCGCG GCAAAACATC AAAACGCCCG 1151 TCAAAATCGC CATCTTCACG CTCATCTGCA CGCAGTTGAT GAACCTCGCC 1201 TTTATCGGTC CGTTGAAACA CGCCGGGCTT TCGCTCGCCA TCGGCCTGGG 1251 CGCGTGCATC AACGCCGGAT TGTTGTTCTT CCTGTTGCGC AAACACGGTA 1301 TTTACCGGCC cggcaggggt tgggcggcgt TCTTGGCGAA AATGCTGCTC 1351 GCGCTCGCCG TGATGTGCGG CGGACTGTGG GCGGCGCAGG CTTGCCTGCC 1401 GTTCGAATGG GCGCACGCCG GCGGAATGCG GAAAGCGGGG CAGCTCTGCA 1451 TCCTGATTGC CGTCGGCGGC GGACTGTATT TCGCATCTCT GGCGGCTTTG 1501 GGCTTCCGTC CGCGCCATTT CAAACGCGTG GAAAGCTGA This encodes the following .amino acid sequence <SEQ ID 122; ORF20ng-l>: 1 MNMLGALAKV GSLTMVSRVL GFVRDTVIAR AFGAGMATDA FFVAFKLPNL 51 LRRVFAEGAF AQAFVPILAE YKETRSKEAT EAFIRHVAGM LSFVLIWTA 101 LGILAAPWVI YVSAPGFTKD ADKFQLSISL LRITFPYILL ISLSSFVGSI 151 LNSYHKFGIP AFTPTFLNIS FIVFALFFVP YFDPPVTALA WAVFVGGILQ 201 LGFQLPWLAK LGFLKLPKLN FKDAAVNRVM KQMAPAILGV SVAQISLVIN 251 TIFASYLQSG SVSWMYYADR MMELRRGVLG AALGTILLPT LSKHSANQDT 301 EQFSALLDWG LRLCMLLTLP AAAGLAVLSF PLVATLFMYR EFTLFDAQMT 351 QHALIAYSFG LIGLIMIKVL ASGFYARQNI KTPVKIAIFT LICTQLMNLA 401 FJEGPLKHAGL SLAIGLGACI NAGLLFFLLR KHGIYRPGRG WAAFLAKMLL 451 ALAVMCGGLW AAQACLPFEW AHAGGMRKAG QLCILIAVGG GLYFASLAAL 501 GFRPRHFKRV ES* ORF20ng-l and ORF20-1 show 95.7% identity in 512 aa overlap: 10 20 30 40 50 60 orf20-1. pep MNMLGALAKVGSLTMVSRVLGFVRDTVIARAFGAGMATDAFFVAFKLPNLLRRVFAEGAF II II I I II I I II I I II I I II I I I I I I I I I I II II II I I II I II II II I I II II II I I I II orf20ng-l MNMLGALAKVGSLTMVSRVLGFVRDTVIARAFGAGMATDAFFVAFKLPNLLRRVFAEGAF 10 20 30 40 50 60 70 80 90 100 110 120 orf20-1. pep AQAFVPILAEYKETRSKEAAEAFIRHVAGMLSFVLVIVTALGILAAPWVIYVSAPGFAQD I I I I II I I I M I M M I I M II II II II II II I IM: I II II I I I I I II I I I I I II I :: I orf20ng-l AQAFVPILAEYKETRSKEATEAFIRHVAGMLSFVLIWTALGILAAPWVIYVSAPGFTKD 70 80 90 100 110 120 130 140 150 160 170 180 orf20-1. pep ADKFQLSIDLLRITFPYILLISLSSFVGSVLNSYHKFGIPAFTPTFLNVSFIVFALFFVP I I II II II : I I II II II II I II II I II II : II I I I II II II II I II II : II I II I I I I II orf20ng-l ADKFQLSISLLRITFPYILLISLSSFVGSILNSYHKFGIPAFTPTFLNISFIVFALFFVP 130 140 150 160 170 180 190 200 210 220 230 240 orf20-l . pep YFDPPVTALAWAVFVGGILQLGFQLPWLAKLGFLKLPKLSFKDAAVNRVMKQMAPAILGV I II II I II II I I I I II II I II I II I II II I I I I II II I I : I II II II I I I II I II II II I orf20ng-l YFDPPVTALAWAVFVGGILQLGFQLPWLAKLGFLKLPKLNFKDAAVNRVMKQMAPAILGV 190 200 210 220 230 240 250 260 270 280 290 300 orf20-1. pep SVAQVSLVINTIFASYLQSGSVSWMYYADRMMELPSGVLGAALGTILLPTLSKHSANQDT II II : II II I I I II I I II I I I I I I II I II II I II I II II II II II II I I I I I II II I I orf20ng-l SVAQISLVINTIFASYLQSGSVSWMYYADRMMELRRGVLGAALGTILLPTLSKHSANQDT 250 260 270 280 290 300 310 320 330 340 350 360 orf20-1. pep EQFSALLDWGLRLCMLLTLPAAVGLAVLSFPLVATLFMYREFTLFDAQMTQHALIAYSFG II II II I I I I II II II II I I II : I I II I I I II I II I II II I II I I I I I I II II I I I I I I I orf20ng-l EQFSALLDWGLRLCMLLTLPAAAGLAVLSFPLVATLFMYREFTLFDAQMTQHALIAYSFG 310 320 330 340 350 360 370 380 390 400 410 420 orf20-1. pep LIGLIMIKVLAPGFYARQNIKTPVKIAIFTLICTQLMNLAFIGPLKHVGLSLAIGLGACI II I II II I I II I II I I I II I II I II I I I II I II I II II II II I I I I : I I I II I I I I I I I orf20ng-l LIGLIMIKVLASGFYARQNIKTPVKIAIFTLICTQLMNLAFIGPLKHAGLSLAIGLGACI 370 380 390 400 410 420 430 440 450 460 470 480 orf20-1. pep NAGLLFYLLRRHGIYQPGKGWAAFLAKMLLSLAVMCGGLWAAQAYLPFEWAHAGGMRKAG II II II : II I : I I I I : I I I I II I II II I M I I I I II II II II I II I I II II I I I I I I I orf20ng-l NAGLLFFLLRKHGIYRPGRGWAAFLAKMLLALAVMCGGLWAAQACLPFEWAHAGGMRKAG 430 440 450 460 470 480 490 500 510 orf20-l.pep QLCILIAVGGGLYFASLAALGFRPRHFKRVENX I II II I I I II II II I I II I I II I II I II I I I : I orf20ng-l QLCILIAVGGGLYFASLAALGFRPRHFKRVESX 490 500 510 ition, ORF20ng-l shows significant homology with a virulence factor of S. typhimurium: sp|P37169|MVIN_SALTY VIRULENCE FACTOR MVIN pιr||S40271 mviN protein - Salmonella typhimurium gi 1438252 (Z26133) mviB gene product [Salmonella typhimurium] gnl|PID|dl005521 (D25292) ORF2 [Salmonella typhimurium] Length = 524 Score = 1573 (750.1 bits), Expect = l.le-220, Sum P(2) = l.le-220 Identities = 309/467 (66%), Positives = 368/467 (78%) Query: 1 MNMLGALAKVGSLTMVSRVLGFVRDTVIARAFGAGMATDAFFVAFKLPNLLRRVFAEGAF 60 MN+L +LA V S+TM SRVLGF RD ++AR FGAGMATDAFFVAFKLPNLLRR+FAEGAF Sbjct: 14 MNLLKSLAAVSSMTMFSRVLGFARDAIVARIFGAGMATDAFFVAFKLPNLLRRIFAEGAF 73 Query: 61 AQAFVPILAEYKETRSKEATEAFIRHVAGMLSFVLIWTALGILAAPWVIYVSAPGFTKD 120 +QAFVPILAEYK + +EAT F+ +V+G+L+ L WT G+LAAPWVI V+APGF Sbjct: 74 SQAFVPILAEYKSKQGEEATRIFVAYVSGLLTLALAWTVAGMLAAPWVIMVTAPGFADT 133 Query: 121 ADKFQLSISLLRITFPYILLISLSSFVGSILNSYHKFGIPAFTPTFLNISFIVFALFFVP 180 ADKF L+ LLRITFPYILLISL+S VG+ILN++++F IPAF PTFLNIΞ I FALF P Sbjct: 134 ADKFALTTQLLRITFPYILLISLASLVGAILNTWNRFSIPAFAPTFLNISMIGFALFAAP 193 Query: 181 YFDPPVTALAWAVFVGGILQLGFQLPWLAKLGFLKLPKLNFKDAAVNRVMKQMAPAILGV 240 YF+PPV ALAWAV VGG+LQL +QLP+L K+G L LP++NF+D RV+KQM PAILGV Sbjct: 194 YFNPPVLALAWAVTVGGVLQLVYQLPYLKKIGMLVLPRINFRDTGAMRWKQMGPAILGV 253 Query: 241 SVAQISLVINTIFASYLQSGSVSWMYYADRMMELRRGVLGAALGTILLPTLSKHSANQDT 300 SV+QISL+INTIFAS+L SGSVSWMYYADR+ME GVLG ALGTILLP+LSK A+ + Sbjct: 254 SVSQISLIINTIFASFLASGSVSWMYYADRLMEFPSGVLGVALGTILLPSLSKSFASGNH 313 Query: 301 EQFSALLDWGLRLCMLLTLPAAAGLAVLSFPLVATLFMYREFTLFDAQMTQHALIAYSFG 360 +++ L+DWGLRLC LL LP+A L +L+ PL +LF Y +FT FDA MTQ ALIAYS G Sbjct: 314 DEYCRLMDWGLRLCFLLALPSAVALGILAKPLTVSLFQYGKFTAFDAAMTQRALIAYSVG 373 Query: 361 LIGLIMIKVLASGFYARQNIKTPVKIAIFTLICTQLMNLAFIGPLKHAGLSLAIGLGACI 420 LIGLI++KVLA GFY+RQ+IKTPVKIAI TLI TQLMNLAFIGPLKHAGLSL+IGL AC+ Sbjct: 374 LIGLIWKVLAPGFYSRQDIKTPVKIAIVTLIMTQLMNLAFIGPLKHAGLSLSIGLAACL 433 Query: 421 NAGLLFFLLRKHGIYRPGRGWXXXXXXXXXXXXVMCGGLWAAQACLP 467 NA LL++ LRK 1+ P GW VM L+ +P Sbjct: 434 NASLLYWQLRKQNIFTPQPGWMWFLMRLIISVLVMAAVLFGVLHIMP 480 Score = 70 (33.4 bits), Expect = l.le-220, Sum P(2) = l.le-220 Identities = 14/41 (34%), Positives = 23/41 (56%) Query: 469 EWAHAGGMRKAGQLCILIAVGGGLYFASLAALGFRPRHFKR 509 EW+ + + +L ++ G YFA+LA LGF+ + F R Sbjct: 481 EWSQGSMLWRLLRLMAWIAGIAAYFAALAVLGFKVKEFVR 521 Based on this analysis, including the homology with a virulence factor from S.typhimurium, it is predicted that these proteins from N. meningitidis and N.gonorrhoeae, and their epitopes, could be useful antigens for vaccines or diagnostics, or for raising antibodies. Example 15 The following partial DNA sequence was identified in N. meningitidis <SEQ ID 123>: 1 atGATTAAAA TCAAAAAAGG TCTAAACCTG CCCATCGCGG GCAGACCGGA 51 GCAAGCCGTT tACGACGGCC CGGCCaTTAC CGAAGtCGCG TTGCTTGGCG 101 AAGAATATGC CGGTATGCGC CCCTCGATGA AAGTCAAGGA AGGCGATGCC 151 GTcAAAAAAG GCCAAGTGCT GTTTGAAGAC AAAAAGAATC CGGGCGTGGT 201 GTTTACTGCG CCGGCTTCAG GcAAAATCGC CGCGATTCAC CGTGGCGAAA 251 AGCGCGTACT TCAGTCAGTC GTGATTGCCG TTGAArGCAA CGACGAAATC 301 GAGTTTGAAC GCTACGCACC TGAAGCGCTG GCAAACTTAA GCGGCGAAGA 351 AGTGCGCCGC AACCTGATCC AATCCGGTTT GTGGACTGCG CTGCGCACCC 401 GTCCGTTCAG CAAAATTCCT GCCGTCGATG CCGAGCCGTT CGCCATCTTC 451 GTCAATGCGA tGGACACCAA TCCG.. This corresponds to the amino acid sequence <SEQ ID 124; ORF22>: 1 MIKIKKGLNL PIAGRPEQAV YDGPAITEVA LLGEEYAGMR PSMKVKEGDA 51 VKKGQVLFED KKNPGWFTA PASGKIAAIH RGEKRVLQSV VIAVEXNDEI 101 EFERYAPEAL ANLSGEEVRR NLIQSGLWTA LRTRPFSKIP AVDAEPFAIF 151 VNAMDTNP.. Further work revealed the complete nucleotide sequence <SEQ ID 125>: 1 ATGATTAAAA TCAAAAAAGG TCTAAACCTG CCCATCGCGG GCAGACCGGA 51 GCAAGCCGTT TACGACGGCC CGGCCATTAC CGAAGTCGCG TTGCTTGGCG 101 AAGAATATGC CGGTATGCGC CCCTCGATGA AAGTCAAGGA AGGCGATGCC 151 GTCAAAAAAG GCCAAGTGCT GTTTGAAGAC AAAAAGAATC CGGGCGTGGT 201 GTTTACTGCG CCGGCTTCAG GCAAAATCGC CGCGATTCAC CGTGGCGAAA 251 AGCGCGTACT TCAGTCAGTC GTGATTGCCG TTGAAGGCAA CGACGAAATC 301 GAGTTTGAAC GCTACGCACC TGAAGCGCTG GCAAACTTAA GCGGCGAAGA 351 AGTGCGCCGC AACCTGATCC AATCCGGTTT GTGGACTGCG CTGCGCACCC 401 GTCCGTTCAG CAAAATTCCT GCCGTCGATG CCGAGCCGTT CGCCATCTTC 451 GTCAATGCGA TGGACACCAA TCCGCTGGCT GCCGACCCTA CGGTCATTAT 501 CAAAGAAGCC GCCGAGGATT TCAAACGCGG CCTGTTGGTA TTGAGCCGTT 551 TGACCGAACG CAAAATCCAT GTTTGTAAGG CAGCTGGCGC AGACGTGCCG 601 TCTGAAAATG CTGCCAACAT CGAAACACAT GAATTCGGCG GCCCGCATCC 651 TGCCGGTTTG AGTGGCACGC ACATTCATTT CATCGAGCCG GTCGGCGCGA 701 ATAAAACCGT GTGGACCATC AATTATCAAG ATGTAATTAC CATTGGCCGT 751 TTGTTTGCAA CAGGCCGTCT GAACACCGAG CGCGTGATTG CCCTAGGTGG 801 TTCTCAAGTC AACAAACCGC GCCTCTTGCG TACCGTTTTG GGTGCGAAAG 851 TATCGCAAAT TACTGCGGGC GAATTGGTTG ACACAGACAA CCGCGTGATT 901 TCCGGTTCGG TATTGAACGG CGCGATTACA CAAGGCGCGC ACGATTATTT 951 GGGACGCTAC CACAATCAGA TTTCCGTTAT CGAAGAAGGC CGCAGCAAAG 1001 AGCTGTTCGG CTGGGTTGCG CCGCAGCCGG ACAAATACTC CATCACGCGT 1051 ACAACCCTCG GCCATTTCCT GAAAAACAAA CTCTTCAAGT TCAACACAGC 1101 CGTCAACGGC GGCGACCGCG CCATGGTGCC GATTGGTACT TACGAGCGCG 1151 TGATGCCCTT GGATATCCTG CCCACCCTGC TTTTGCGCGA TTTAATCGTC 1201 GGCGATACCG ACAGCGCGCA GGCATTGGGT TGCTTGGAAT TGGACGAAGA 1251 AGACCTCGCT TTGTGCAGCT TCGTCTGCCC GGGCAAATAC GAATACGGCC 1301 CGCTGTTGCG CAAAGTGCTG GAAACCATTG AGAAGGAAGG CTGA This corresponds to the amino acid sequence <SEQ ID 126; ORF22-l>: 1 MIKIKKGLNL PIAGRPEQAV YDGPAITEVA LLGEEYAGMR PSMKVKEGDA 51 VKKGQVLFED KKNPGWFTA PASGKIAAIH RGEKRVLQSV VIAVEGNDEI 101 EFERYAPEAL ANLSGEEVRR NLIQSGLWTA LRTRPFSKIP AVDAEPFAIF 151 VNAMDTNPLA ADPTVIIKEA AEDFKRGLLV LSRLTERKIH VCKAAGADVP 201 SENAANIETH EFGGPHPAGL SGTHIHFIEP VGANKTVWTI NYQDVITIGR 251 LFATGRLNTE RVIALGGSQV NKPRLLRTVL GAKVSQITAG ELVDTDNRVI 301 SGSVLNGAIT QGAHDYLGRY HNQISVIEEG RSKELFGWVA PQPDKYSITR 351 TTLGHFLKNK LFKFNTAVNG GDRAMVPIGT YERVMPLDIL PTLLLRDLIV 401 GDTDSAQALG CLELDEEDLA LCSFVCPGKY EYGPLLRKVL ETIEKEG* Further work identified the corresponding gene in strain A of N. meningitidis <SEQ D 127>: 1 ATGATTAAAA TCAAAAAAGG TCTAAACCTG CCCATCGCGG GCAGACCGGA 51 GCAAGTCATT TATGACGGGC CCGTCATTAC CGAAGTCGCG TTGCTTGGCG 101 AAGAATATGC CGGTATGCGC CCCTNGATGA AAGTCAAGGA AGGCGATGCC 151 GTCAAAAAAG GCCAAGTGCT GTTTGAAGAC AAAAAGNATC CGGGCGTGGT 201 GTTTACCGCG CCNGTTTCAG GCAAAATCGC CGCCATCCAT CGCGGCGAAA 251 AGCGCGTACT TCAGTCGGTC GTGATTGCCG TTGAAGGCAA CGACGAAATC 301 GAGTTCGAAC GCTACGCGCC CGAAGCGTTG GCAAACTTAA GCGGCGANGA 351 ANTNNGNNGC AATCTGATCC AATCCGGTTT GTGGACTGCG CTGCGTANCC 401 GTCCGTTCAG CAAAATCCCT GCCGTCGATG CCGAGCCGTT CGCCATCTTC 451 GTCAATGCGA TGGACACCAA TCCGCTNGCG GCAGACCCTG TGGTTGTGAT 501 CAAAGAAGCC GNCGANGATT TCAGACGANG TNTGCTGGTA TTGAGCCGTT 551 TGACCGAGCG TAAAATCCAT GTGTGTAAGG CAGCTGGCGC AGACGTGCCG 601 TCTGAAAATG CTGCCAACAT CGAAACACAT GAATTCGGCG GCCCGCATCC 651 GGCCGGTTTG AGTGGCACGC ACATTCATTT CATTGAGCCG GTCGGTGCAA 701 ACAAAACCGT TTGGACCATC AATTATCAAG ATGTAATTGC CATCGGACGT 751 TTGTTTGCAA CAGGCCGTCT GAACACCGAG CGCGTGATTG CTTTGGGTGG 801 TTCTCAAGTC AACAAACCAC GCCTCTTGCG TACCGTTTTG GGTGCGAAAG 851 TATCGCAAAT TACTGCGGGC GAATTGGTTG ACGCAGACAA CCGCGTGATT 901 TCCGGTTCGG TATTGAACGG CGCGATTACA CAAGGCGCGC ACGATTATTT 951 GGGACGCTAC CACAATCAGA TTTCCGTTAT CGAAGAAGGC CGCAGCAAAG 1001 AGCTGTTCGG CTGGGTTGCG CCGCAGCCGG ACAAATACTC CATCACGCGT 1051 ACGACCCTCG GCCATTTCCT GAAAAACAAA CTCTTCAAGT TCACGACAGC 1101 CGTCAACGGT GGCGACCGCG CCATGGTGCC GATTGGTACT TACGAGCGCG 1151 TAATGCCGCT AGACATCCTG CCTACCCTGC TTTTGCGCGA TTTAATCGTC 1201 GGCGATACCG ACAGCGCGCA AGCATTGGGT TGCTTGGAAT TGGACGAAGA 1251 AGACCTCGCT TTGTGCAGCT TCGTCTGCCC GGGCAAATAC GAATANGGCC 1301 CGCTGTTGCG TAAGGTGCTG GAAACCNTTG AGAAGGAAGG CTGA This encodes a protein having amino acid sequence <SEQ LD 128; ORF22a>: 1 MIKIKKGLNL PIAGRPEQVI YDGPVITEVA LLGEEYAGMR PXMKVKEGDA 51 VKKGQVLFED KKXPGWFTA PVSGKIAAIH RGEKRVLQSV VIAVEGNDEI 101 EFERYAPEAL ANLSGXEXXX NLIQSGLWTA LRXRPFSKIP AVDAEPFAIF 151 VNAMDTNPLA ADPWVIKEA XXDFRRXXLV LSRLTERKIH VCKAAGADVP 201 SENAANIETH EFGGPHPAGL SGTHIHFIEP VGANKTVWTI NYQDVIAIGR 251 LFATGRLNTE RVIALGGSQV NKPRLLRTVL GAKVSQITAG ELVDADNRVI 301 SGSVLNGAIT QGAHDYLGRY HNQISVIEEG RSKELFGWVA PQPDKYSITR 351 TTLGHFLKNK LFKFTTAVNG GDRAMVPIGT YERVMPLDIL PTLLLRDLIV 401 GDTDSAQALG CLELDEEDLA LCSFVCPGKY EXGPLLRKVL ETXEKEG* The originally-identified partial strain B sequence (ORF22) shows 94.2% identity over a 158aa overlap with ORF22a: 10 20 30 40 50 60 orf22. pep MIKIKKGLNLPIAGRPEQAVYDGPAITEVALLGEEYAGMRPSMKVKEGDAVKKGQVLFED I II I I II II I I I I II I I I :: I I I I: I II I II II II I I I I I I I I II II II I II II II I II orf22a MIKIKKGLNLPIAGRPEQVIYDGPVITEVALLGEEYAGMRPXMKVKEGDAVKKGQVLFED 10 20 30 40 50 60 70 80 90 100 110 120 orf22. pep KKNPGWFTAPASGKIAAIHRGEKRVLQSWIAVEXNDEIEFERYAPEALANLSGEEVRR II II II I I I M II I I II II I I I I I I I I I I I I I I I I II I II II II I I I I II I I I I orf 22a KKXPGWFTAPVSGKIAAIHRGEKRVLQSWIAVEGNDEIEFERYAPEALANLSGXEXXX 70 80 90 100 110 120 130 140 150 orf22 . pep NLIQSGLWTALRTRPFSKIPAVDAEPFAIFVNAMDTNP I I I I I I I I I I I I : I I I I I I I I I I I I I I I I I I I I I I I I I orf 22a NLIQSGLWTALRXRPFSKIPAVDAEPFAIFVNAMDTNPLAADPWVIKEAXXDFRRXXLV 130 140 150 160 170 180 The complete strain B sequence (ORF22-1) and ORF22a show 94.9% identity in 447 aa overlap: 10 20 30 40 50 60 orf22a. pep MIKIKKGLNLPIAGRPEQVIYDGPVITEVALLGEEYAGMRPXMKVKEGDAVKKGQVLFED I II II I II I I I II II II I :: I I II : II II II I I II II II I I II II I II I I I I I I I II I I o 22-1 MIKIKKGLNLPIAGRPEQAVYDGPAITEVALLGEEYAGMRPSMKVKEGDAVKKGQVLFED 10 20 30 40 50 60 70 80 90 100 110 120 orf 22a . pep KKXPGWFTAPVSGKIAAIHRGEKRVLQSWIAVEGNDEIEFERYAPEALANLSGXEXXX I I I I I I I I I M I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I orf 22-1 KKNPGWFTAPASGKIAAIHRGEKRVLQSWIAVEGNDEIEFERYAPEALANLSGEEVRR 70 80 90 100 110 120 130 140 150 160 170 180 orf 22a . pep NLIQSGLWTALRXRPFSKIPAVDAEPFAIFVNAMDTNPLAADPVWIKEAXXDFRRXXLV I II II I II II II: I II II II I I II I II II II I II I II I II II M I : II I I IM I II orf22-1 NLIQSGLWTALRTRPFSKIPAVDAEPFAIFVNAMDTNPLAADPTVIIKEAAEDFKRGLLV 130 140 150 160 170 180 190 200 210 220 230 240 orf22a. pep LSRLTERKIHVCKAAGADVPSENAANIETHEFGGPHPAGLSGTHIHFIEPVGANKTVWTI II I II II I II I I II II II II I II I I II II I I I II I II I II II I I II II I I I II II I I I I I or 22-1 LSRLTERKIHVCKAAGADVPSENAANIETHEFGGPHPAGLSGTHIHFIEPVGANKTVWTI 190 200 210 220 230 240 250 260 270 280 290 300 orf22a . pep NYQDVIAIGRLFATGRLNTERVIALGGSQVNKPRLLRTVLGAKVSQITAGELVDADNRVI I II II I : I II I I I I I I I II I II I II I II II II I II I II I II II II I I I II II I I: I II I I orf22-1 NYQDVITIGRLFATGRLNTERVIALGGSQVNKPRLLRTVLGAKVSQITAGELVDTDNRVI 250 260 270 280 290 300 310 320 330 340 350 360 orf22a . pep SGSVLNGAITQGAHDYLGRYHNQISVIEEGRSKELFGWVAPQPDKYSITRTTLGHFLKNK I I I I I II I I II II II I II I I I II I I II II II I II I II I II I I I I II I I I II II I I I I I I I orf22-1 SGSVLNGAITQGAHDYLGRYHNQISVIEEGRSKELFGWVAPQPDKYSITRTTLGHFLKNK 310 320 330 340 350 360 370 380 390 400 410 420 orf22a . pep LFKFTTAVNGGDRAMVPIGTYERVMPLDILPTLLLRDLIVGDTDSAQALGCLELDEEDLA I I II: II II II II I II II I I II II I II I I I II I I I I I II II I I II II I I I II I I I I II I I orf22-1 LFKFNTAVNGGDRAMVPIGTYERVMPLDILPTLLLRDLIVGDTDSAQALGCLELDEEDLA 370 380 390 400 410 420 430 440 orf22a . pep LCSFVCPGKYEXGPLLRKVLETXEKEGX I I II I I I II I I II I I I II I II II I II orf22-l LCSFVCPGKYEYGPLLRKVLETIEKEGX 430 440 Further work identified a partial gene sequence <SEQ LD 129> from N.gonorrhoeae, which encodes the following amino acid sequence <SEQ ID 130; ORF22ng>: 1 MIKIKKGLNL PIAGRPEQVI YDGPAITEVA LLGEEYVGMR PSMKIKEGEA 51 VKKGQVLFED KKNPGWFTA PASGKIAAIH RGEKRVLQSV VIAVEGNDEI 101 EFERYVPEAL AKLSSEKVRR NLIQSGLWTA LRTRPFSKIP AVDAEPFAIF 151 VNAMDTNPLA ADPTVIIKEA AEDFKRGLLV LSRLTERKIH VCKAAGADVP 201 SENAANIETH EFGGPHPAGL SGTHIHFIEP VGANKTVWTI NYQDVIAIGR 251 LFVTGRLNTE RWALGGLQV NKPRLLRTVL GAKVSQLTAG ELVDADNRVI 301 SGSVLNGAIA QGAHDYLGRY HN* Further work identified complete gonococcal gene <SEQ ID 131 > • 1 ATGATTAAAA TCAAAAAAGG TCTAAATCTG CCCATCGCGG GCAGACCGGA 51 GCAAGTCATT TATGACGGCC CGGCCATTAC CGAAGTCGCG TTGCTTGGCG 101 AAGAATATGT CGGCATGCGC CCCTCGATGA AAATCAAGGA AGGTGAAGCC 151 GTCAAAAAAG GCCAAGTGCT GTTTGAAGAC AAAAAGAATC CGGGCGTAGT 201 ATTTACTGCG CCGGCTTCAG GCAAAATCGC CGCTATTCAC CGTGGCGAAA 251 AGCGCGTACT TCAGTCAGTC GTGATTGCCG TTGAAGGCAA CGACGAAATC 301 GAGTTCGAAC GCTACGTACC TGAAGCGCTG GCAAAATTGA GCAGCGAAAA 351 AGTGCGCCGC AACCTGATTC AATCAGGCTT ATGGACTGCG CTTCGCACCC 401 GTCCGTTCAG CAAAATCCCT GCCGTAGATG CCGAGCCGTT CGCCATCTTC 451 GTCAATGCGA TGGACACCAA TCCGCTGGCT GCCGACCCTA CGGTCATCAT 501 CAAAGAAGCC GCCGAAGACT TCAAACGCGG CCTGTTGGTA TTGAGCCGCC 551 TGACCGAACG TAAAATCCAT GTGTGTAAAG CAGCAGGCGC AGACGTGCCG 601 TCTGAAAATG CTGCCAATAT CGAAACACAT GAATTTGGCG GCCCGCATCC 651 TGCCGGCTTG AGTGGCACGC ACATTCATTT CATCGAGCCA GTCGGCGCGA 701 ATAAAACCGT GTGGACCATC AATTATCAAG ACGTGATTGC TATCGGACGT 751 TTGTTCGTAA CAGGCCGTCT GAATACCGAG CGCGTGGTTG CCTTGGGCGG 801 CCTGCAAGTC AACAAACCGC GCCTCTTGCG TACCGTTTTG GGTGCGAAGG 851 TGTCTCAACT TACCGCCGGC GAATTGGTTG ACGCGGACAA CCGCGTGATT 901 TCCGGTTCGG TATTGAACGG TGCGATTGCA CAAGGCGCGC ATGATTATTT 951 GGGACGCTAC CACAATCAGA TTTCCGTTAT CGAAGAAGGC CGCAGCAAAG 1001 AGCTGTTCGG CTGGGTTGCG CCGCAGCCGG ACAAATACTC CATCACGCGC 1051 ACCACTCTCG GCCATTTCCT AAAAAACAAA CTCTTCAAGT TCACGACAGC 1101 CGTCAACGGC GGCGACCGCG CCATGGTACC GATCGGCACT TATGAGCGCG 1151 TAATGCCGTT GGACATCCTG CCTACCTTGC TTTTGCGCGA TTTAATCGTC 1201 GGCGATACCG ACAGCGCGCA GGCTTTGGGT TGCTTGGAAT TGGACGAAGA 1251 AGACCTCGCT TTGTGCAGCT TCGTCTGCCC GGGCAAATAC GAATACGGCC 1301 CGCTGTTGCG CAAAGTGCTG GAAACCATTG AGAAGGAAGG CTGA This encodes a protein having amino acid sequence <SEQ ID 132; ORF22ng-l>: 1 MIKIKKGLNL PIAGRPEQVI YDGPAITEVA LLGEEYVGMR PSMKIKEGEA 51 VKKGQVLFED KKNPGWFTA PASGKIAAIH RGEKRVLQSV VIAVEGNDEI 101 EFERYVPEAL AKLSSEKVRR NLIQSGLWTA LRTRPFSKIP AVDAEPFAIF 151 VNAMDTNPLA ADPTVIIKEA AEDFKRGLLV LSRLTERKIH VCKAAGADVP 201 SENAANIETH EFGGPHPAGL SGTHIHFIEP VGANKTVWTI NYQDVIAIGR 251 LFVTGRLNTE RWALGGLQV NKPRLLRTVL GAKVSQLTAG ELVDADNRVI 301 SGSVLNGAIA QGAHDYLGRY HNQISVIEEG RSKELFGWVA PQPDKYSITR 351 TTLGHFLKNK LFKFTTAVNG GDRAMVPIGT YERVMPLDIL PTLLLRDLIV 401 GDTDSAQALG CLELDEEDLA LCSFVCPGKY EYGPLLRKVL ETIEKEG* The originally-identified partial strain B sequence (ORF22) shows 93.7% identity over a 158aa overlap with ORF22ng: orf22. pep MIKIKKGLNLPIAGRPEQAVYDGPAITEVALLGEEYAGMRPSMKVKEGDAVKKGQVLFED 60 II I I II I I II I I II I I I M: I I I I II I I I I II I I IM I II I I I MM M II I II I I I II I orf22ng MIKIKKGLNLPIAGRPEQVIYDGPAITEVALLGEEYVGMRPSMKIKEGEAVKKGQVLFED 60 orf22.pep KKNPGWFTAPASGKIAAIHRGEKRVLQSWIAVEXNDEIEFERYAPEALANLSGEEVRR 120 I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I : I I I I I : I M I : I I I orf22ng KKNPGVVFTAPASGKIAAIHRGEKRVLQSWIAVEGNDEIEFERYVPEALAKLSSEKVRR 120 orf22.pep NLIQSGLWTALRTRPFSKIPAVDAEPFAIFVNAMDTNP 158 I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I orf22ng NLIQSGLWTALRTRPFSKIPAVDAEPFAIFVNAMDTNPLAADPTVIIKEAAEDFKRGLLV 180 The complete sequences from strain B (ORF22-1) and gonococcus (ORF22ng) show 96.2% identity in 447 aa overlap: 10 20 30 40 50 60 orf22-1. pep MIKIKKGLNLPIAGRPEQAVYDGPAITEVALLGEEYAGMRPSMKVKEGDAVKKGQVLFED I II II II II I I I II I I I I :: I I I II I II II I I II I I : II I II I I : II I : II I I I I I I I I I orf22ng-l MIKIKKGLNLPIAGRPEQVIYDGPAITEVALLGEEYVGMRPSMKIKEGEAVKKGQVLFED 10 20 30 40 50 60 70 80 90 100 110 120 orf22-1. pep KKNPGWFTAPASGKIAAIHRGEKRVLQSWIAVEGNDEIEFERYAPEALANLSGEEVRR I II II II I I I II I II II I I I I II II I II I II I II I II II II I II I : I II IM IM M I II orf22ng-l KKNPGWFTAPASGKIAAIHRGEKRVLQSWIAVEGNDEIEFERYVPEALAKLSSEKVRR 70 80 90 100 110 120 130 140 150 160 170 180 orf22-1. pep NLIQSGLWTALRTRPFSKIPAVDAEPFAIFVNAMDTNPLAADPTVIIKEAAEDFKRGLLV I I I I II II II II I II I I II I I I I I II II I I I I I II II II II I II II I II II I II II II II orf22ng-l NLIQSGLWTALRTRPFSKIPAVDAEPFAIFVNAMDTNPLAADPTVIIKEAAEDFKRGLLV 130 140 150 160 170 180 190 200 210 220 230 240 orf22-1. pep LSRLTERKIHVCKAAGADVPSENAANIETHEFGGPHPAGLSGTHIHFIEPVGANKTVWTI II II I I I I II I II I I II I I I I II I II II I II I I II I I I II I II II II II I II I I I I I II I orf22ng-l LSRLTERKIHVCKAAGADVPSENAANIETHEFGGPHPAGLSGTHIHFIEPVGANKTVWTI 190 200 210 220 230 240 250 260 270 280 290 300 orf22-1. pep NYQDVITIGRLFATGRLNTERVIALGGSQVNKPRLLRTVLGAKVSQITAGELVDTDNRVI I II II I : II I II : I II I II I I I : II I I I I I I I II II I II I I I I I M I I I II I I : I I I II orf22ng-l NYQDVIAIGRLFVTGRLNTERWALGGLQVNKPRLLRTVLGAKVSQLTAGELVDADNRVI 250 260 270 280 290 300 310 320 330 340 350 360 orf22-1. pep SGSVLNGAITQGAHDYLGRYHNQISVIEEGRSKELFGWVAPQPDKYSITRTTLGHFLKNK I I II I I II M II II I I I I I I II II II I I II I I I II II II II I II I I II I I II I I I I I I II orf22ng-l SGSVLNGAIAQGAHDYLGRYHNQISVIEEGRSKELFGWVAPQPDKYSITRTTLGHFLKNK 310 320 330 340 350 360 370 380 390 400 410 420 orf22-1. pep LFKFNTAVNGGDRAMVPIGTYERVMPLDILPTLLLRDLIVGDTDSAQALGCLELDEEDLA I I I I : II II I I II I I I I I II I I I I I II I I I I II I I I I II II I II II II II II I I I I I II I orf22ng-l LFKFTTAVNGGDRAMVPIGTYERVMPLDILPTLLLRDLIVGDTDSAQALGCLELDEEDLA 370 380 390 400 410 420 430 440 orf22-1. pep LCSFVCPGKYEYGPLLRKVLETIEKEGX II II I I II II II II II I II I I I I I I I I I orf22ng-l LCSFVCPGKYEYGPLLRKVLETIEKEGX 430 440 Computer analysis of these sequences gave the following results: Homology with 48kDa outer membrane protein of Actinobacillus pleuropneumoniae (accession number U24492). ORF22 and this 48kDa protein show 72% aa identity in 158aa overlap: Orf22 1 MIKIKKGLNLPIAGRPEQAVYDGPAITEVALLGEEYAGMRPSMKVKEGDAVKKGQVLFED 60 MI IKKGL+LPIAG P Q +++G + EVA+LGEEY GMRPSMKV+EGD VKKGQVLFED 48kDa 1 MITIKKGLDLPIAGTPAQVIHNGNTVNEVAMLGEEYVGMRPSMKVREGDWKKGQVLFED 60 orf22 61 KKNPGWFTAPASGKIAAIHRGEKRVLQSWIAVEXNDEIEFERYAPEALANLSGEEVRR 120 KKNPGWFTAPASG + I+RGEKRVLQSWI VE +++I F RY LA+LS E+V++ 48kDa 61 KKNPGWFTAPASGTWTINRGEKRVLQSWIKVEGDEQITFTRYEAAQLASLSAEQVKQ 120 orf22 121 NLIQSGLWTALRTRPFSKIPAVDAEPFAIFVNAMDTNP 158 NLI+SGLWTA RTRPFSK+PA+DA P +IFVNAMDTNP 48kDa 121 NLIESGLWTAFRTRPFSKVPALDAIPSSIFVNAMDTNP 158 ORF22a also shows homology to the 48kDa Actinobacillus pleuropneumoniae protein: gi 11185395 (U24492) 48 kDa outer membrane protein [Actinobacillus pleuropneumoniae] Length = 449 Score = 530 bits (1351), Expect = e-150 Identities = 274/450 (60%), Positives = 323/450 (70%), Gaps = 4/450 (0%) Query: 1 MIKIKKGLNLPIAGRPEQVIYDGPVITEVALLGEEYAGMRPXMKVKEGDAVKKGQVLFED 60 MI IKKGL+LPIAG P QVI++G + EVA+LGEEY GMRP MKV+EGD VKKGQVLFED Sbjct: 1 MITIKKGLDLPIAGTPAQVIHNGNTVNEVAMLGEEYVGMRPSMKVREGDWKKGQVLFED 60 Query: 61 KKXPGWFTAPVSGKIAAIHRGEKRVLQSWIAVEGNDEIEFERYAPEALANLSGXEXXX 120 KK PGWFTAP SG + I+RGEKRVLQSWI VEG+++I F RY LA+LS + Sbjct: 61 KKNPGWFTAPASGTWTINRGEKRVLQSWIKVEGDEQITFTRYEAAQLASLSAEQVKQ 120 Query: 121 NLIQSGLWTALRXRPFSKIPAVDAEPFAIFVNAMDTNPLAADPVWIKEAXXDFRRXXLV 180 NLI+SGLWTA R RPFSK+PA+DA P +IFVNAMDTNPLAADP W+KE DF+ V Sbjct: 121 NLIESGLWTAFRTRPFSKVPALDAIPSSIFVNAMDTNPLAADPEWLKEYETDFKDGLTV 180 Query: 181 LSRL—TERKIHVCKAAGADVP-SENAANIETHEFGGPHPAGLSGTHIHFIEPVGANKTV 237 L+RL ++ +++CK A +++P S I F G HPAGL GTHIHF++PVGA K V Sbjct: 181 LTRLFNGQKPVYLCKDADSNIPLSPAIEGITIKSFSGVHPAGLVGTHIHFVDPVGATKQV 240 Query: 238 WTINYQDVIAIGRLFATGRLNTERVIALGGSQVNKPRLLRTVLGAKVSQITAGELVDADN 297 W +NYQDVIAIG+LF TG L T+R+I+L G QV PRL+RT LGA +SQ+TA EL +N Sbjct: 241 WHLNYQDVIAIGKLFTTGELFTDRIISLAGPQVKNPRLVRTRLGANLSQLTANELNAGEN 300 Query: 298 RVISGSVLNGAITQGAHDYLGRYHNQISVIEEGRSKELFGWVAPQPDKYSITRTTLGHFL 357 RVISGSVL+GA G DYLGRY Q+SV+ EGR KELFGW+ P DK+SITRT LGHF Sbjct: 301 RVISGSVLSGATAAGPVDYLGRYALQVSVLAEGREKELFGWIMPGSDKFSITRTVLGHFG 360 Query: 358 KNKLFKFTTAVNGGDRAMVPIGTYERVMXXXXXXXXXXXXXXVGDTDSAQXXXXXXXXXX 417 K KLF FTTAV+GG+RAMVPIG YERVM GDTDSAQ Sbjct: 361 K-KLFNFTTAVHGGERAMVPIGAYERVMPLDIIPTLLLRDLAAGDTDSAQNLGCLELDEE 419 Query: 418 XXXXXSFVCPGKYEXGPLLRKVLETXEKEG 447 ++VCPGK GP+LR LE EKEG ORF22ng-l also shows homology with the OMP from A.pleuropneumoniae: gi 11185395 (U24492) 48 kDa outer membrane protein [Actinobacillus pleuropneumoniae] Length = 449 Score = 555 bits (1414), Expect = e-157 Identities = 284/450 (63%), Positives = 337/450 (74%), Gaps = 4/450 (0%) Query: 27 MIKIKKGLNLPIAGRPEQVIYDGPAITEVALLGEEYVGMRPSMKIKEGEAVKKGQVLFED 86 MI IKKGL+LPIAG P QVI++G + EVA+LGEEYVGMRPSMK++EG+ VKKGQVLFED Sbjct: 1 MITIKKGLDLPIAGTPAQVIHNGNTVNEVAMLGEEYVGMRPSMKVREGDWKKGQVLFED 60 Query: 87 KKNPGWFTAPASGKIAAIHRGEKRVLQSWIAVEGNDEIEFERYVPEALAKLSSEKVRR 146 KKNPGWFTAPASG + I+RGEKRVLQSWI VEG+++I F RY LA LS+E+V++ Sbjct: 61 KKNPGWFTAPASGTWTINRGEKRVLQSWIKVEGDEQITFTRYEAAQLASLSAEQVKQ 120 Query: 147 NLIQSGLWTALRTRPFSKIPAVDAEPFAIFVNAMDTNPLAADPTVIIKEAAEDFKRGLLV 206 NLI+SGLWTA RTRPFSK+PA+DA P +IFVNAMDTNPLAADP V++KE DFK GL V Sbjct: 121 NLIESGLWTAFRTRPFSKVPALDAIPSSIFVNAMDTNPLAADPEWLKEYETDFKDGLTV 180 Query: 207 LSRL—TERKIHVCKAAGADVP-SENAANIETHEFGGPHPAGLSGTHIHFIEPVGANKTV 263 L+RL ++ +++CK A +++P S I F G HPAGL GTHIHF++PVGA K V Sbjct: 181 LTRLFNGQKPVYLCKDADSNIPLSPAIEGITIKSFSGVHPAGLVGTHIHFVDPVGATKQV 240 Query: 264 WTINYQDVIAIGRLFVTGRLNTERWALGGLQVNKPRLLRTVLGAKVSQLTAGELVDADN 323 W +NYQDVIAIG+LF TG L T+R+++L G QV PRL+RT LGA +SQLTA EL +N Sbjct: 241 WHLNYQDVIAIGKLFTTGELFTDRIISLAGPQVKNPRLVRTRLGANLSQLTANELNAGEN 300 Query: 324 RVISGSVLNGAIAQGAHDYLGRYHNQISVIEEGRSKELFGWVAPQPDKYSITRTTLGHFL 383 RVISGSVL+GA A G DYLGRY Q+SV+ EGR KELFGW+ P DK+SITRT LGHF Sbjct: 301 RVISGSVLSGATAAGPVDYLGRYALQVSVLAEGREKELFGWIMPGSDKFSITRTVLGHFG 360 Query: 384 KNKLFKFTTAVNGGDRAMVPIGTYERVMXXXXXXXXXXXXXXVGDTDSAQXXXXXXXXXX 443 K KLF FTTAV+GG+RAMVPIG YERVM GDTDSAQ Sbjct: 361 K-KLFNFTTAVHGGERAMVPIGAYERVMPLDIIPTLLLRDLAAGDTDSAQNLGCLELDEE 419 Query: 444 XXXXXSFVCPGKYEYGPLLRKVLETIEKEG 473 ++VCPGK YGP+LR LE IEKEG Sbjct: 420 DLALCTYVCPGKNNYGPMLRAALEKIEKEG 449 Based on this analysis, including the homology with the outer membrane protein of Actinobacillus pleuropneumoniae, it was predicted that these proteins from N. meningitidis and N.gonorrhoeae, and their epitopes, could be useful antigens for vaccines or diagnostics, or for raising antibodies. ORF22-1 (35.4kDa) was cloned in pET and pGex vectors and expressed in E.coli, as described above. The products of protein expression and purification were analyzed by SDS-PAGE. Figure 5 A shows the results of affinity purification of the GST- fusion protein, and Figure 5B shows the results of expression of the His-fusion in E.coli. Purified GST-fusion protein was used to immunise mice, whose sera were used for ELISA (positive result) and FACS analysis (Figure 5C). These experiments confirm that ORF22-1 is a surface-exposed protein, and that it is a useful immunogen. Example 16 The following partial DNA sequence was identified in N. meningitidis <SEQ ID 133>: 1 .. GCGnCGnAAA TCATCCATCC CC .. nACGTC GTAGGCCCTG AAGCCAACTG 51 GTTTTTTATG GTAGCCAGTA CGTTTGTGAT TGCTTTGATT GGTTATTTTG 101 TTACTGAAAA AATCGTCGAA CCGCAATTGG GCCCTTATCA ATCAGATTTG 151 TCACAAGAAG AAAAAGACAT TCGGCATTCC AATGAAATCA CGCCTTTGGA 201 ATATAAAGGA TTAATTTGGG CTGGCGTGGT GTTTGTTGCC TTATCCGCCC 251 TATTGGCTTG GAGCATCGTC CCTGCCGACG GTATTTTGCG TCATCCTGAA 301 ACAGGATTGG TTTCCGGTTC GCCGTTTTTA AAATCGATTG TTGTTTTTAT 351 TTTCTTGTTG TTTGCACTGC CGGGCATTGT TTATGGCCGG GTAACCCGAA 401 GTTTGCGCGG CGAACAGGAA GTCGTTAATG CGmyGGCCGA ATCGATGAGT 451 ACTCTGGsGC TTTmTTTGsw CAkcATCTTT TTTGCCGCAC AGTTTGTCGC 501 ATTTTTTAAT TGGACGAATA TTGGGCAATA TATTGCCGTT AAAGGGGCGA 551 CGTTCTTAAA AGAAGTCGGC TTGGGCGGCA GCGTGTTGTT TATCGGTTTT 601 ATTTTAATTT GTGCTTTTAT CAATCTGATG ATAGGCTCCG CCTCCGCGCA 651 ATGGGCGGTA ACTGCGCCGA TTTTCGTCCC TATGCTGATG TTGGCCGGCT 701 ACGCGCCCGA AGTCATTCAA GCCGCTTACC GCATCGGTGA TTCCGTTACC 751 AATATTATTA CGCCGATGAT GAGTTATTTC GGGCTGATTA TGGCGACGGT 801 GrkCmmmTAC AAAAAAGATG CGGGCGTGGG TaCGcTGATT wCTATGATGT 851 TGCCGTATTC CGCTTTCTTC TTGATTGCgT GGATTGCCTT ATTCTGCATT 901 TGGGTATTTg TTTTGGGCCT GCCCGTCGGT CCCGGCGCGC CCACATTCTA 951 TCCCGCACCT TAA This corresponds to the .amino acid sequence <SEQ ID 134; ORF12>: 1 ..AXXIIHPXXV VGPEANWFFM VASTFVIALI GYFVTEKIVE PQLGPYQSDL 51 SQEEKDIRHS NEITPLEYKG LIWAGWFVA LSALLAWSIV PADGILRHPE 101 TGLVSGSPFL KSI FIFLL FALPGIVYGR VTRSLRGEQE WNAXAESMS 151 TLXLXLXXIF FAAQFVAFFN WTNIGQYIAV KGATFLKEVG LGGSVLFIGF 201 ILICAFINLM IGSASAQWAV TAPIFVPMLM LAGYAPEVIQ AAYRIGDSVT 251 NIITPMMSYF GLIMATVXXY KKDAGVGTLI XMMLPYSAFF LIAWIALFCI 301 WVFVLGLPVG PGAPTFYPAP * Further sequence analysis revealed the complete DNA sequence <SEQ LD 135> to be: 1 ATGAGTCAAA CCGATACGCA ACGGGACGGA CGATTTTTAC GCACAGTCGA 51 ATGGCTGGGC AATATGTTGC CGCATCCGGT TACGCTTTTT ATTATTTTCA 101 TTGTGTTATT GCTGATTGCC TCTGCCGTCG GTGCGTATTT CGGACTATCC 151 GTCCCCGATC CGCGCCCTGT TGGTGCGAAA GGACGTGCCG ATGACGGTTT 201 GATTTACATT GTCAGCCTGC TCAATGCCGA CGGTTTTATC AAAATCCTGA 251 CGCATACCGT TAAAAATTTC ACCGGTTTCG CGCCGTTGGG AACGGTGTTG 301 GTTTCTTTAT TGGGCGTGGG GATTGCGGAA AAATCGGGCT TGATTTCCGC 351 ATTAATGCGC TTATTGCTCA CAAAATCGCC ACGCAAACTC ACTACTTTTA 401 TGGTTGTTTT TACAGGGATT TTATCTAATA CCGCTTCTGA ATTGGGCTAT 451 GTCGTCCTAA TCCCTTTGTC CGCCATCATC TTTCATTCCC TCGGCCGCCA 501 TCCGCTTGCC GGTCTGGCTG CGGCTTTCGC CGGCGTTTCG GGCGGTTATT 551 CGGCCAATCT GTTCTTAGGC ACAATCGATC CGCTCTTGGC AGGCATCACC 601 CAACAGGCGG CGCAAATCAT CCATCCCGAC TACGTCGTAG GCCCTGAAGC 651 CAACTGGTTT TTTATGGTAG CCAGTACGTT TGTGATTGCT TTGATTGGTT 701 ATTTTGTTAC TGAAAAAATC GTCGAACCGC AATTGGGCCC TTATCAATCA 751 GATTTGTCAC AAGAAGAAAA AGACATTCGG CATTCCAATG AAATCACGCC 801 TTTGGAATAT AAAGGATTAA TTTGGGCTGG CGTGGTGTTT GTTGCCTTAT 851 CCGCCCTATT GGCTTGGAGC ATCGTCCCTG CCGACGGTAT TTTGCGTCAT 901 CCTGAAACAG GATTGGTTTC CGGTTCGCCG TTTTTAAAAT CGATTGTTGT 951 TTTTATTTTC TTGTTGTTTG CACTGCCGGG CATTGTTTAT GGCCGGGTAA 1001 CCCGAAGTTT GCGCGGCGAA CAGGAAGTCG TTAATGCGAT GGCCGAATCG 1051 ATGAGTACTC TGGGGCTTTA TTTGGTCATC ATCTTTTTTG CCGCACAGTT 1101 TGTCGCATTT TTTAATTGGA CGAATATTGG GCAATATATT GCCGTTAAAG 1151 GGGCGACGTT CTTAAAAGAA GTCGGCTTGG GCGGCAGCGT GTTGTTTATC 1201 GGTTTTATTT TAATTTGTGC TTTTATCAAT CTGATGATAG GCTCCGCCTC 1251 CGCGCAATGG GCGGTAACTG CGCCGATTTT CGTCCCTATG CTGATGTTGG 1301 CCGGCTACGC GCCCGAAGTC ATTCAAGCCG CTTACCGCAT CGGTGATTCC 1351 GTTACCAATA TTATTACGCC GATGATGAGT TATTTCGGGC TGATTATGGC 1401 GACGGTGATC AAATACAAAA AAGATGCGGG CGTGGGTACG CTGATTTCTA 1451 TGATGTTGCC GTATTCCGCT TTCTTCTTGA TTGCGTGGAT TGCCTTATTC 1501 TGCATTTGGG TATTTGTTTT GGGCCTGCCC GTCGGTCCCG GCGCGCCCAC 1551 ATTCTATCCC GCACCTTAA This corresponds to the amino acid sequence <SEQ ID 136; ORF12-l>: 1 MSQTDTQRDG RFLRTVEWLG NMLPHPVTLF IIFIVLLLIA SAVGAYFGLS 51 VPDPRPVGAK GRADDGLIYI VSLLNADGFI KILTHTVKNF TGFAPLGTVL 101 VSLLGVGIAE KSGLISALMR LLLTKSPRKL TTFMWFTGI LSNTASELGY 151 WLIPLSAII FHSLGRHPLA GLAAAFAGVS GGYSANLFLG TIDPLLAGIT 201 QQAAQIIHPD YWGPEANWF_ FMVASTFVIA LIGYFVTEKI VEPQLGPYQS 251 DLSQEEKDIR HSNEITPLEY KGLIWAGWF VALSALLAWS IVPADGILRH 301 PETGLVSGSP FLKSIWFIF LLFALPGIVY GRVTRSLRGE QEWNAMAES 351 MSTLGLYLVI IFFAAQFVAF FNWTNIGQYI AVKGATFLKE VGLGGSVLFI 401 GFILICAFIN LMIGSASAQW AVTAPIFVPM LMLAGYAPEV IQAAYRIGDS 451 VTNIITPMMS YFGLIMATVI KYKKDAGVGT LISMMLPYSA FFLIAWIALF 501 CIWVFVLGLP VGPGAPTFYP AP* Computer analysis of this amino acid sequence gave the following results: Homology with a predicted ORF from N.meninsitidis (strain A) ORF12 shows 96.3% identity over a 320aa overlap with an ORF (ORF12a) from strain A of N meningitidis: 10 20 30 orf 12 . pep AXXI IHPXXWGPEAΝWFFMVASTFVIALI I I I I I I I I I I I I I I I I I I I I I I I I I I orf 12a AAAFAGVSGGYSAΝLFLGTIDPLLAGITQQAAQIIHPDYWGPEAΝWFFMVASTFVIALI 180 190 200 210 220 230 40 50 60 70 80 90 orf12. pep GYFVTEKIVEPQLGPYQSDLSQEEKDIRHSΝEITPLEYKGLIWAGWFVALSALLAWSIV I I I I I II I I I I I II I II II I I I I II I II II I I I I I I II I I I I I I II I II I II II II II M orf12a GYFVTEKIVEPQLGPYQSDLSQEEKDIRHSΝEITPLEYKGLIWAGWFVALSALLAWSIV 240 250 260 270 280 290 100 110 120 130 140 150 orf12. pep PADGILRHPETGLVΞGSPFLKSIWFIFLLFALPGIVYGRVTRSLRGEQEWΝAXAESMS I I I I I II I II I I II I II I II II I I I I II I I I II I II I I I II I I II II I I II I I I II II I orf12a PADGILRHPETGLVSGSPFLKSIWFIFLLFALPGIVYGRVTRSLRGEQEWΝAMAESMS 300 310 320 330 340 350 160 170 180 190 200 210 orf12. pep TLXLXLXXIFFAAQFVAFFΝWTΝIGQYIAVKGATFLKEVGLGGSVLFIGFILICAFIΝLM II I I I I II I I I II II I II I I I I II II I II I I I II I I I I I I II I I I I I I II I II II I orf12a TLGLYLVIIFFAAQFVAFFΝWTΝIGQYIAVKGATFLKEVGLGGSVLFIGFILICAFIΝLM 360 370 380 390 400 410 220 230 240 250 260 270 orf12. pep IGSASAQWAVTAPIFVPMLMLAGYAPEVIQAAYRIGDSVTΝIITPMMSYFGLIMATVXXY I II I I I I II II I I I II I II I II I I I I II II I I I I I I II I I I I I I II I I I II I II I M I orfl2a IGSASAQWAVTAPIFVPMLMLAGYAPEVIQAAYRIGDSVTNIITPMMSYFGLIMATVIKY 420 430 440 450 460 470 280 290 300 310 320 orf12. pep KKDAGVGTLIXMMLPYSAFFLIAWIALFCIWVFVLGLPVGPGAPTFYPAPX I II I I II I II II I II II II I II I I II I I I II I I I I I I II I I I II I I II I I orf12a KKDAGVGTLISMMLPYSAFFLIAWIALFCIWVFVLGLPVGPGAPTFYPAPX 480 490 500 510 520 The complete length ORF12a nucleotide sequence <SEQ ID 137> is: 1 ATGAGTCAAA CCGATACGCA ACGGGACGGA CGATTTTTAC GCACAGTCGA 51 ATGGCTGGGC AATATGTTGC CGCACCCGGT TACGCTTTTT ATTATTTTCA 101 TTGTGTTATT GCTGATTGCC TCTGCCGCCG GTGCGTATTT CGGACTATCC 151 GTCCCCGATC CGCGCCCTGT TGGTGCGAAA GGACGTGCCG ATGACGGTTT 201 GATTCACGTT GTCAGCCTGC TCGATGCTGA CGGTTTGATC AAAATCCTGA 251 CGCATACCGT TAAAAATTTC ACCGGTTTCG CGCCGTTGGG AACGGTGTTG 301 GTTTCTTTAT TGGGCGTGGG GATTGCGGAA AAATCGGGCT TGATTTCCGC 351 ATTAATGCGC TTATTGCTCA CAAAATCTCC ACGCAAACTC ACTACTTTTA 401 TGGTTGTTTT TACAGGGATT TTATCTAATA CCGCTTCTGA ATTGGGCTAT 451 GTCGTCCTAA TCCCTTTGTC CGCCATCATC TTTCATTCCC TCGGCCGCCA 501 TCCGCTTGCC GGTCTGGCTG CGGCTTTCGC CGGCGTTTCG GGCGGTTATT 551 CGGCCAATCT GTTCTTAGGC ACAATCGATC CGCTCTTGGC AGGCATCACC 601 CAACAGGCGG CGCAAATCAT CCATCCCGAC TACGTCGTAG GCCCTGAAGC 651 CAACTGGTTT TTTATGGTAG CCAGTACGTT TGTGATTGCT TTGATTGGTT 701 ATTTTGTTAC TGAAAAAATC GTCGAACCGC AATTGGGCCC TTATCAATCA 751 GATTTGTCAC AAGAAGAAAA AGACATTCGA CATTCCAATG AAATCACGCC 801 TTTGGAATAT AAAGGATTAA TTTGGGCTGG CGTGGTGTTT GTTGCCTTAT 851 CCGCCCTATT GGCTTGGAGC ATCGTCCCTG CCGACGGTAT TTTGCGTCAT 901 CCTGAAACAG GATTGGTTTC CGGTTCGCCG TTTTTAAAAT CAATTGTTGT 951 TTTTATTTTC TTGTTGTTTG CACTGCCGGG CATTGTTTAT GGCCGGGTAA 1001 CCCGAAGTTT GCGCGGCGAA CAGGAAGTCG TTAATGCGAT GGCCGAATCG 1051 ATGAGTACTC TGGGGCTTTA TTTGGTCATC ATCTTTTTTG CCGCACAGTT 1101 TGTCGCATTT TTTAATTGGA CGAATATTGG GCAATATATT GCCGTTAAAG 1151 GGGCGACGTT CTTAAAAGAA GTCGGCTTGG GCGGCAGCGT GTTGTTTATC 1201 GGTTTTATTT TAATTTGTGC TTTTATCAAT CTGATGATAG GCTCCGCCTC 1251 CGCGCAATGG GCGGTAACTG CGCCGATTTT CGTCCCTATG CTGATGTTGG 1301 CCGGCTACGC GCCCGAAGTC ATTCAAGCCG CTTACCGCAT CGGTGATTCC 1351 GTTACCAATA TTATTACGCC GATGATGAGT TATTTCGGGC TGATTATGGC 1401 GACGGTGATC AAATACAAAA AAGATGCGGG CGTGGGTACG CTGATTTCTA 1451 TGATGTTGCC GTATTCCGCT TTCTTCTTGA TTGCGTGGAT TGCCTTATTC 1501 TGCATTTGGG TATTTGTTTT GGGCCTGCCC GTCGGTCCCG GCGCGCCCAC 1551 ATTCTATCCC GCACCTTAA This encodes a protein having amino acid sequence <SEQ ID 138>: 1 MSQTDTQRDG RFLRTVEWLG NMLPHPVTLF IIFIVLLLIA SAAGAYFGLS 51 VPDPRPVGAK GRADDGLIHV VSLLDADGLI KILTHTVKNF TGFAPLGTVL 101 VSLLGVGIAE KSGLISALMR LLLTKSPRKL TTFMWFTGI LSNTASELGY 151 WLIPLSAII FHSLGRHPLA GLAAAFAGVS GGYSANLFLG TIDPLLAGIT 201 QQAAQIIHPD YWGPEANWF FMVASTFVIA LIGYFVTEKI VEPQLGPYQS 251 DLSQEEKDIR HSNEITPLEY KGLIWAGWF VALSALLAWS IVPADGILRH 301 PETGLVSGSP FLKSIWFIF LLFALPGIVY GRVTRSLRGE QEWNAMAES 351 MSTLGLYLVI IFFAAQFVAF FNWTNIGQYI AVKGATFLKE VGLGGSVLFI 401 GFILICAFIN LMIGSASAQW AVTAPIFVPM LMLAGYAPEV IQAAYRIGDS 451 VTNIITPMMS YFGLIMATVI KYKKDAGVGT LISMMLPYSA FFLIAWIALF 501 CIWVFVLGLP VGPGAPTFYP AP* ORF12a and ORF12-1 show 99.0% identity in 522 aa overlap: 10 20 30 40 50 60 orf12a . pep MSQTDTQRDGRFLRTVEWLGNMLPHPVTLFIIFIVLLLIASAAGAYFGLSVPDPRPVGAK I I I II I I II I II I II I II I I II I I I I II I II I II II II I I I I : I I I I I I I I I I I I II II I orf12-1 MSQTDTQRDGRFLRTVEWLGNMLPHPVTLFIIFIVLLLIASAVGAYFGLSVPDPRPVGAK 10 20 30 40 50 60 70 80 90 100 110 120 orf 12a . pep GRADDGLIHWSLLDADGLIKILTHTVKNFTGFAPLGTVLVSLLGVGIAEKSGLISALMR I I I I I I I M M ! I M l I M I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I orf 12-1 GRADDGLIYIVSLLNADGFIKILTHTVKNFTGFAPLGTVLVSLLGVGIAEKSGLISALMR 70 80 90 100 110 120 130 140 150 160 170 180 orf12a. pep LLLTKSPRKLTTFMWFTGILSNTASELGYWLIPLSAIIFHSLGRHPLAGLAAAFAGVS II II II II II II II II I II II I I I I II I II II I II I II I I I I II I II I II I II II I I I I I orf12-1 LLLTKSPRKLTTFMWFTGILSNTASELGYWLIPLSAIIFHSLGRHPLAGLAAAFAGVS 130 140 150 160 170 180 190 200 210 220 230 240 orf12a . pep GGYSANLFLGTIDPLLAGITQQAAQIIHPDYWGPEANWFFMVASTFVIALIGYFVTEKI II II II I I I II I I II I II II I I II II I I I I I I II I I II I I II I I I II I II I II II I I I I I orf12-1 GGYSANLFLGTIDPLLAGITQQAAQIIHPDYWGPEANWFFMVASTFVIALIGYFVTEKI 190 200 210 220 230 240 250 260 270 280 290 300 orf12a . pep VEPQLGPYQSDLSQEEKDIRHSNEITPLEYKGLIWAGWFVALSALLAWSIVPADGILRH I I I II I I II I I II I I I II II I I II II I I II I I II I I II II I I II I I I II I II I I II I I II orf12-1 VEPQLGPYQSDLSQEEKDIRHSNEITPLEYKGLIWAGWFVALSALLAWSIVPADGILRH 250 260 270 280 290 300 310 320 330 340 350 360 orf12a.pep PETGLVSGSPFLKSIWFIFLLFALPGIVYGRVTRSLRGEQEWNAMAESMSTLGLYLVI II I II I I II II I II I I I I I I II I II I I II I I I II I I I I II II I I I II I II II II II I II I orf12-1 PETGLVSGSPFLKSIWFIFLLFALPGIVYGRVTRSLRGEQEWNAMAESMSTLGLYLVI 310 320 330 340 350 360 370 380 390 400 410 420 orf12a. pep IFFAAQFVAFFNWTNIGQYIAVKGATFLKEVGLGGSVLFIGFILICAFINLMIGSASAQW I II I II II II II I I I I I I II I I II I II I I I I II I II I II II II I II II II I II II I I I I I orf12-1 IFFAAQFVAFFNWTNIGQYIAVKGATFLKEVGLGGSVLFIGFILICAFINLMIGSASAQW 370 380 390 400 410 420 430 440 450 460 470 480 orf12a. pep AVTAPIFVPMLMLAGYAPEVIQAAYRIGDSVTNIITPMMSYFGLIMATVIKYKKDAGVGT I I I II I I II II I I I I I I I I I I I II I II I I I II I II I I II I I I II I I I II II I I I I I I I I I orf12-1 AVTAPIFVPMLMLAGYAPEVIQAAYRIGDSVTNIITPMMSYFGLIMATVIKYKKDAGVGT 430 440 450 460 470 480 490 500 510 520 orf12a. pep LISMMLPYSAFFLIAWIALFCIWVFVLGLPVGPGAPTFYPAPX I II I II II I I II I I I II II I I I I II I I II II I II I II I II II I orf12-1 LISMMLPYSAFFLIAWIALFCIWVFVLGLPVGPGAPTFYPAPX 490 500 510 520 Homology with a predicted ORF from N.sonorrhoeae ORF12 shows 92.5% identity over a 320aa overlap with a predicted ORF (ORF12.ng) from N. gonorrhoeae: orf12. pep AXXIIHPXXWGPEANWFFMVASTFVIALI 30 I II I I I II II I I II I I : II II I I I I I orfl2ng AAAFAGVSGGYSANLFLGTIDPLLAGITQQAAQIIHPDYVVGPEANWFFMAASTFVIALI 232 orf12.pep GYFVTEKIVEPQLGPYQSDLSQEEKDIRHSNEITPLEYKGLIWAGWFVALSALLAWΞIV 90 II II II II II I II II II I I I I I I I I I I I I II II I II I I I II II II I II II II I I II I II I orfl2ng GYFVTEKIVEPQLGPYQSDLSQEEKDIRHSNEITPLEYKGLIWAGWFVALSALLAWSIV 292 orf12. pep PADGILRHPETGLVSGSPFLKSIWFIFLLFALPGIVYGRVTRSLRGEQEWNAXAEΞMS 150 I I II I I II I I I I I I: I I I I II I I I I I I II II II I I II I IM I I II II I : I II II I II I I orfl2ng PADGILRHPETGLVAGSPFLKSIWFIFLLFALPGIVYGRITRSLRGEREWNAMAESMS 352 orf12. pep TLXLXLXXIFFAAQFVAFFNWTNIGQYIAVKGATFLKEVGLGGSVLFIGFILICAFINLM 210 II I I II I I I I I I II I II I I II I I II I I I M I I I : II I I I I I II II II II I I I I I orfl2ng TLGLYLVIIFFAAQFVAFFNWTNIGQYIAVKGAVFLKKFRLGGSVLFIGFILICAFINLM 412 orf12.pep IGSASAQWAVTAPIFVPMLMLAGYAPEVIQAAYRIGDSVTNIITPMMSYFGLIMATVXXY 270 M II I I II I II I II I I M I II I I I I : II I I I I II I I I I I I I II I I I II II II II I I I orfl2ng IGSASAQWAVTAPIFVPMLMLAGNAPQVIQAAYRIGDSVTNIITPMMSYFGLIMATVIKY 472 orfl2.pep KKDAGVGTLIXMMLPYSAFFLIAWIALFCIWVFVLGLPVGPGAPTFYPAP 320 II I II I II II I I I I I II II I I I I I II I I I II I II I I I I II M II I I I : I orfl2ng KKDAGVGTLISMMLPYSAFFLIAWIALFCIWVFVLGLPVGPGTPTFYPVP 522 The complete length ORF12ng nucleotide sequence <SEQ ID 139> is: 1 ATGAGTCAAA CCGACGCGCG TCGTAGCGGA CGATTTTTAC GCACAGTCGA 51 ATGGCTGGGC AATATGTTGC CGCACCCGGT TACGCTTTTT ATTATTTTCA 101 TTGTGTTATT GCTGATTGcc tctgCCGTCG GTGCGTATTT CGGACTATCC 151 GTCCCCGATC CGCGTCCTGT TGGGGCGAAA GGACGTGCCG ATGACGGTTT 201 GATTCACGTT GTCAGCCTGC TCGATGCCGA CGGTTTGATC AAAATCCTGA 251 CGCATACCGT TAAAAATTTC ACCGGTTTCG CGCCGTTGGG AACGGTGTTG 301 GTTTCTTTAT TGGGCGTGGG GATTGCGGAA AAATCGGGCT TGATTTCCGC 351 ATTAATGCGC TTATTGCTCA CAAAATCCCC ACGCAAACTC ACTACTTTTA 401 TGGTTGTTTT TACAGGGATT TTATCCAATA CGGCTTCTGA ATTGGGCTAT 451 GTCGTCCTAA TCCCTTTGTC CGCCGTCATC TTTCATTCGC TCGGCCGCCA 501 TCCGCTTGCC GGTTTGGCTG CGGCTTTCGC CGGCGTTTCG GGCGGTTATT 551 CGGCCAATCT GTTCTTAGGC ACAATCGATC CGCTCTTGGC AGGCATCACC 601 CAACAGGCGG CGCAAATCAT CCATCCCGAC TACGTCGTAG GCCCTGAAGC 651 CAACTGGTTT TTTATGGCAG CCAGTACGTT TGTGATTGCT TTGATTGGTT 701 ATTTTGTTAC TGAAAAAATC GTCGAACCGC AATTGGGCCC TTATCAATCA 751 GATTTGTCAC AAGAAGAAAA AGACATTCGG CATTCCAATG AAATCACGCC 801 TTTGGAATAT AAAGGATTAA TTTGGGCAGG CGTGGTGTTT GTTGCCTTAT 851 CCGCCCTATT GGCTTGGAGC ATCGTCCCTG CCGACGGTAT TTTGCGTCAT 901 CCTGAAACAG GATTGGTTGC CGGTTCGCCG TTTTTAAAAT CGATTGTTGT 951 TTTTATTTTC TTGTTGTTTG CGCTGCCGGG CATTGTTTAT GGCCGGATAA 1001 CCCGAAGTTT GCGCGGCGAA CGGGAAGTCG TTAATGCGAT GGCCGAATCG 1051 ATGAGTACTT TGGGACTTTA TTTGGTCATC ATCTTTTTTG CCGCACAGTT 1101 TGTCGCATTT TTTAATTGGA CGAATATTGG GCAATATATT GCCGTTAAAG 1151 GGGCGGTGTT CTTAAAAGAA GTCGGCTTGG GCGGCAGTGT GTTGTTTATC 1201 GGTTTTATTT TAATTTGTGC TTTTATCAAT CTGATGATAG GCTCCGCCTC 1251 CGCGCAATGG GCGGTAACTG CGCCGATTTT CGTCCCTATG CTGATGTTGG 1301 CCGGCTACGC GCCCGAAGTC ATTCAAGCCG CTTACCGCAT CGGTGATTCC 1351 GTTACCAATA TTATTACGCC GATGATGAGT TATTTCGGGC TGATTATGGC 1401 GACGGTAATC AAATACAAAA AAGATGCGGG CGTAGGCACG CTGATTTCTA 1451 TGATGTTGCC GTATTCCGCT TTCTTCTTAA TTGCATGGAT CGCCTTATTC 1501 TGCATTTGGG TATTTGTTTT GGGTCTGCCC GTCGGTCCCG GCACACCCAC 1551 ATTCTATCCG GTGCCTTAA This encodes a protein having amino acid sequence <SEQ ID 140>: 1 MSQTDARRSG RFLRTVEWLG NMLPHPVTLF IIFIVLLLIA SAVGAYFGLS 51 VPDPRPVGAK GRADDGLIHV VSLLDADGLI KILTHTVKNF TGFAPLGTVL 101 VSLLGVGIAE KSGLISALMR LLLTKSPRKL TTFMWFTGI LSNTASELGY 151 VVLIPLSAVI FHSLGRHPLA GLAAAFAGVS GGYSANLFLG TIDPLLAGIT 201 QQAAQIIHPD YVVGPEANWF FMAASTFVIA LIGYFVTEKI VEPQLGPYQS 251 DLSQEEKDIR HSNEITPLEY KGLIWAGWF VALSALLAWS IVPADGILRH 301 PETGLVAGSP FLKSI FIF LLFALPGIVY GRITRSLRGE REWNAMAES 351 MSTLGLYLVI IFFAAQ VAF FNWTNIGQYI AVKGAVFLKK FRLGGSVLFI 401 GFILICAFIN LMIGSASAQW AVTAPIFVPM LMLAGNAPQV IQAAYRIGDS 451 VTNIITPMMS YFGLIMATVI KYKKDAGVGT LISMMLPYSA FFLIAWIALF 501 CIWVFVLGLP VGPGTPTFYP VP* ORF12ng shows 97.1% identity in 522 aa overlap with ORF 12-1: 10 20 30 40 50 60 orf12-1. pep MSQTDTQRDGRFLRTVEWLGNMLPHPVTLFIIFIVLLLIASAVGAYFGLSVPDPRPVGAK orfl2ng MSQTDARRSGRFLRTVEWLGNMLPHPVTLFIIFIVLLLIASAVGAYFGLSVPDPRPVGAK 10 20 30 40 50 60 70 80 90 100 110 120 orf 12-1 . pep GRADDGLIYIVSLLNADGFIKILTHTVKNFTGFAPLGTVLVSLLGVGIAEKSGLISALMR I I I I I I I I : : I I I M I I M I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I orfl2ng GRADDGLIHWSLLDADGLIKILTHTVKNFTGFAPLGTVLVSLLGVGIAEKSGLISALMR 70 80 90 100 110 120 130 140 150 160 170 180 orf 12-1 . pep LLLTKSPRKLTTFMWFTGILSNTASELGYWLIPLSAIIFHSLGRHPLAGLAAAFAGVS I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I : I I I I I I I I I I I I I I I I I I I I I orf l2ng LLLTKSPRKLTTFMWFTGILSNTASELGYWLIPLSAVIFHSLGRHPLAGLAAAFAGVS 130 140 150 160 170 180 190 200 210 220 230 240 orf12-1. pep GGYSANLFLGTIDPLLAGITQQAAQIIHPDYWGPEANWFFMVASTFVIALIGYFVTEKI II II I I II II II II I I II I I II II I II I I I I I II I I I I II I I : II I I I I I I II II I II I I orfl2ng GGYSANLFLGTIDPLLAGITQQAAQIIHPDYWGPEANWFFMAASTFVIALIGYFVTEKI 190 200 210 220 230 240 250 260 270 280 290 300 orf12-1. pep VEPQLGPYQSDLSQEEKDIRHSNEITPLEYKGLIWAGVVFVALSALLAWSIVPADGILRH I II II II I I I I II II I II II II II I II I II II II I II I II II II I I I I I II I I II I I I II orfl2ng VEPQLGPYQSDLSQEEKDIRHSNEITPLEYKGLIWAGWFVALSALLAWSIVPADGILRH 250 260 270 280 290 300 310 320 330 340 350 360 orf12-1. pep PETGLVSGSPFLKSIWFIFLLFALPGIVYGRVTRSLRGEQEWNAMAESMSTLGLYLVI I I II IM I I I I I I I I I I II I I II I I I I II II I: I II II IMI I II I I I I II I I II I I I II orfl2ng PETGLVAGSPFLKSIWFIFLLFALPGIVYGRITRSLRGEREWNAMAESMSTLGLYLVI 310 320 330 340 350 360 370 380 390 400 410 420 orf12-1. pep IFFAAQFVAFFNWTNIGQYIAVKGATFLKEVGLGGSVLFIGFILICAFINLMIGSASAQW II I I I I II I I I II II I II I I II I II : I I II II I II I II I II I I II I I I I II I I II I I I II orfl2ng IFFAAQFVAFFNWTNIGQYIAVKGAVFLKEVGLGGSVLFIGFILICAFINLMIGSASAQW 370 380 390 400 410 420 430 440 450 460 470 480 orf12-1. pep AVTAPIFVPMLMLAGYAPEVIQAAYRIGDSVTNIITPMMSYFGLIMATVIKYKKDAGVGT I II II II I I I I I I I I I I II I I II I I I I II I II I I I II II I II II II II I I I II II I I II I orfl2ng AVTAPIFVPMLMLAGYAPEVIQAAYRIGDSVTNIITPMMSYFGLIMATVIKYKKDAGVGT 430 440 450 460 470 480 490 500 510 520 orf12-1. pep LISMMLPYSAFFLIAWIALFCIWVFVLGLPVGPGAPTFYPAPX II II I I II I II II I II II I I II I I II I II II I II : I I I I I : I I orfl2ng LISMMLPYSAFFLIAWIALFCIWVFVLGLPVGPGTPTFYPVPX 490 500 510 520 ition, ORF12ng shows significant homology with a hypotehtical protein from E.coli: sp | P46133 | YDAH_EC0LI HYPOTHETICAL 55 . 1 KD PROTEIN IN OGT-DBPA INTERGENIC REGION >gi 1 1787597 (AE000231 ) hypothetical protein in ogt 5 ' region [Escherichia coli ] Length = 510 Score = 329 bits ( 835 ) , Expect = 2e-89 Identities = 178 /507 ( 35% ) , Positives = 281 /507 ( 55% ) , Gaps = 15/507 ( 2 % ) Query : 8 RSGRFLRTVEWLGNMLPHPVTXXXXXXXXXXXASAVGAYFGLSVPDPRPVGAKGRADDGL 67 +SG+ VE +GN +PHP +A+ + FG+S +P D Sbj ct : 13 QSGKLYGWVERIGNKVPHPFLLFIYLIIVLMVTTAILSAFGVSAKNP TDGTP 64 Query : 68 IHWSLLDADGLIKILTHTVKNFTGFAPXXXXXXXXXXXXIAEKSGLISALMRLLLTKSP 127 + V +LL +GL L + +KNF+GFAP +AE+ GL+ ALM + + Sbj ct : 65 WVKNLLSVEGLHWFLPNVIKNFSGFAPLGAILALVLGAGLAERVGLLPALMVKMASHVN 124 Query : 128 RKLTTFMWFTGILSNTASELGYWLIPLSAVIFHSLGRHPLAGLAAAFAGVSGGYSANL 187 + ++MV+F S+ +S+ V++ P+ A+IF ++GRHP+AGL AA AGV G++ANL Sbj ct : 125 ARYASYMVLFIAFFSHISSDAALVIMPPMGALIFLAVGRHPVAGLLAAIAGVGCGFTANL 184 Query : 188 FLGTIDPLLAGITQQAAQIIHPDYWGPEANWFFMAASTFVIALIGYFVTEKIVEPQLGP 247 + T D LL+GI+ +AA +P V NW+FMA+S V+ ++G +T+KI+EP+LG Sbj ct : 185 LIVTTDVLLSGISTEAAAAFNPQMHVSVIDNWYFMASSVWLTIVGGLITDKIIEPRLGQ 244 Query : 248 YQSDLSQEEKDIRHSNEITPLEYKGLIWAGWFVALSALLAWSIVPADGILRHPETGLVA 307 +Q + ++ + + S GL AGVV + A +A ++P +GILR P V Sbj ct : 245 WQGNSDEKLQTLTESQRF GLRIAGWSLLFIAAIALMVIPQNGILRDPINHTVM 298 Query : 308 GSPFLKSIWFIFLLFALPGIVYGRITRSLRGEREWNAMAESMSTLGLYLXXXXXXXXX 367 SPF+K IV I L F + + YG TR++R + ++ + M E M + ++ Sbj ct : 299 PSPFIKGIVPLIILFFFWSLAYGIATRTIRRQADLPHLMIEPMKEMAGFIVMVFPLAQF 358 Query : 368 XXXXNWTNIGQYIAVKGAVFLKEVGLGGSVLFIGFILICAFINLMIGSASAQWAVTAPIF 427 NW+N+G++IAV L+ GL G F+G L+ +F+ + I S SA W++ APIF Sbjct: 359 VAMFNWSNMGKFIAVGLTDILESSGLSGIPAFVGLALLSSFLCMFIASGSAIWSILAPIF 418 Query: 428 VPMLMLAGYAPEVIQAAYRIGDSVTNIITPMMSYFGLIMATVIKYKKDAGVGTLISMMLP 487 VPM ML G+ P Q +RI DS + P+ + L + + +YK DA +GT S++LP Sbjct: 419 VPMFMLLGFHPAFAQILFRIADSSVLPLAPVSPFVPLFLGFLQRYKPDAKLGTYYSLVLP 478 Query: 488 YSAFFLIAWIALFCIWVFVLGLPVGPG 514 Y FL+ W+ + W +++GLP+GPG Sbjct: 479 YPLIFLWWLLMLLAW-YLVGLPIGPG 504 Based on this analysis, including the presence of several putative transmembrane domains and the predicted actinin-type actin-binding domain signature (shown in bold) in the gonococcal protein, it is predicted that the proteins from N. meningitidis .and N.gonorrhoeae, and their epitopes, could be useful antigens for vaccines or diagnostics, or for raising antibodies. Example 17 The following partial DNA sequence was identified in N. meningitidis <SEQ ID 141>: 1 ..ACAGCCGGCG CAGCAGGTTn CnCGGTCTTC GTTTTCGTAA CGGACAGTCA 51 GGTGGAGGTG TTCGGGAACA TCCAGACCGC AGTGGAAACA GGTTTTTTTC 101 ATGGCATTTC GGTTTCGTCT GTGTTTGGTG CGGCGGCACA AGACTCGGCA 151 ATgGCTTCGC GCAGTGCGTC TATACCGGTA TTTTCAGCAA CGGAAATGCG 201 GACGGcGgCA ATTTTTCCCG CAGCGTCGCG CCATATGCCC GTGTTTTgTT 251 CTTCAGACGG CAGCAGGTCG GTTTTGTTGT ACACCTTgAT GCACGGAaTA 301 TCGCCGGCAT GGATTTCTTG CAGTACGTTT TCCACGTCTT CAATCTGCTG 351 TCCGCTGTTC GGAGCGGCGG CATCGACGAC GTGCAGCAGC ACATCgGcTT 401 gCGCGGTTTC TTCCAGCGTG GCgGAAAAGG CGGAAATCAG TTTgTGCGGC 451 agATyGCTnA CGAATCCGAC GGTATCGGTC AGGATAATGC TGCATTCGGG 501 ACT.. This corresponds to the .amino acid sequence <SEQ ID 142; ORF14>: 1 ..TAGAAGXXVF VFVTDSQVEV FGNIQTAVET GFFHGISVSS VFGAAAQDSA 51 MASRSASIPV FSATEMRTAA IFPAASRHMP VFCSSDGSRS VLLYTLMHGI 101 SPAWISCSTF STSSICCPLF GAAASTTCSS TSACAVSSSV AEKAEIΞLCG 151 RXLTNPTVSV RIMLHSG .. Computer analysis of this amino acid sequence gave the following results: Homology with a predicted ORF from N.meninsitidis (strain A) ORF14 shows 94.0% identity over a 167aa overlap with an ORF (ORF14a) from strain A of N. meningitidis: 10 20 30 orf14.pep TAGAAGXXVFVFVTDSQVEVFGNIQTAVET MMM I I II I II: M: II I I : I I II I orf14a GRQLGFLRVGGALFVITAQARVNNALCDCLTTGAAGFAVFVFVTDGQMQVFGNVQPAVET 150 160 170 180 190 200 40 50 60 70 80 90 orf14. pep GFFHGISVSSVFGAAAQDSAMASRSASIPVFSATEMRTAAIFPAASRHMPVFCSSDGSRS I I I I II II II I II I II I I I I I II I II I I I I I I I I I II I I I I II II I I II II II II II I I orf14a GFFHGISVSSVFGAAAQYSAMASRSASIPVFSATEMRTAAIFPAASRHMPVFCSSDGSRS 210 220 230 240 250 260 100 110 120 130 140 150 orf14.pep VLLYTLMHGISPAWISCSTFSTSSICCPLFGAAASTTCSSTSACAVSSSVAEKAEISLCG I I II II I I II I II I I II I II I II I I II I II I I II I II I II I II II I II II I II II I III I orf14a VLLYTLMHGISPAWISCSTFSTSSICCPLFGAAASTTCSSTSACAVSSSVAEKAEISLCG 270 280 290 300 310 320 160 orf14. pep RXLTNPTVSVRIMLHSG I I II I II I I I I I I II I orf14a RSLTNPTVSVRIMLHSGLMYSRRAVVSSVAKSWSFAYMPDLVSRLNRLDLPTLVX 330 340 350 360 370 380 The complete length ORF 14a nucleotide sequence <SEQ ID 143> is: 1 ATGGAGGATT TGCAGGAAAT CGGGTTCGAT GTCGCCGCCG TAAAGGTAGG 51 TCGGCAGCGC GAACATCATC GTCTGCATCA TCCCCAGCCC GGCAACGGCG 101 AGGCGGACGA TGTATTGTTT GCGTTCTTTT TGGTTGGCGG CTTCGATTTT 151 TTGCGCGTCA TAGGGTGCGG CGGTGTAGCC TATCTGCCTG ATTTTCAACA 201 GAATGTCGGA AAGGCGGATT TTGCCGTCGT CCCAGACGAC GCGGCAGCGG 251 TGCGTGCTGT AATTGAGGTC GATGCGGACG ATGCCGTCTG TACGCAAAAG 301 CTGCTGTTCG ATCAGCCAGA CGCAGGCGGC GCAGGTGATG CCGCCGAGCA 351 TTAAAACCGC CTCGCGCGTG CCGCCGTGGG TTTCCACAAA GTCGGACTGG 401 ACTTCGGGCA GGTCGTACAG GCGGATTTGG TCGAGGATTT CTTGGGGCGG 451 CAGCTCGGTT TTTTGCGCGT CGGCGGTGCG TTGTTTGTAA TAACTGCCCA 501 AGCCCGCGTC AATAATGCTT TGTGCGACTG CCTGACAACC GGCGCAGCAG 551 GTTTCGCGGT CTTCGTTTTC GTAACGGACG GTCAGATGCA GGTTTTCGGG 601 AACGTCCAGC CCGCAGTGGA AACAGGTTTT TTTCATGGCA TTTCGGTTTC 651 GTCTGTGTTT GGTGCGGCGG CACAATACTC GGCAATGGCT TCGCGCAGTG 701 CGTCTATACC GGTATTTTCA GCAACGGAAA TGCGGACGGC GGCAATTTTT 751 CCCGCAGCGT CGCGCCATAT GCCCGTGTTT TGTTCTTCAG ACGGCAGCAG 801 GTCGGTTTTG TTGTACACCT TGATGCACGG AATATCGCCG GCATGGATTT 851 CTTGCAGTAC GTTTTCCACG TCTTCAATCT GCTGTCCGCT GTTCGGAGCG 901 GCGGCATCGA CGACGTGCAG CAGCACATCG GCTTGCGCGG TTTCTTCCAG 951 CGTGGCGGAA AAGGCGGAAA TCAGTTTGTG CGGCAGATCG CTGACGAATC 001 CGACGGTATC GGTCAGGATA ATGCTGCATT CGGGACTGAT GTACAGCCGC 051 CGCGCCGTCG TGTCGAGTGT GGCGAAAAGC TGGTCTTTCG CATATATGCC 101 CGACTTGGTC AGCCGGTTGA ACAGACTGGA TTTGCCGACA TTGGTATAG This encodes a protein having amino acid sequence <SEQ ID 144>: 1 MEDLQEIGFD VAAVKVGRQR EHHRLHHPQP GNGEADDVLF AFFLVGGFDF 51 LRVIGCGGVA YLPDFQQNVG KADFAWPDD AAAVRAVIEV DADDAVCTQK 101 LLFDQPDAGG AGDAAEH*NR LARAAVGFHK VGLDFGQWQ ADLVEDFLGR 151 QLGFLRVGGA LFVITAQARV NNALCDCLTT GAAGFAVFVF VTDGQMQVFG 201 NVQPAVETGF FHGISVSSVF GAAAQYSAMA SRSASIPVFS ATEMRTAAIF 251 PAASRHMPVF CSSDGSRSVL LYTLMHGISP AWISCSTFST SSICCPLFGA 301 AASTTCSSTS ACAVSSSVAE KAEISLCGRS LTNPTVSVRI MLHSGLMYSR 351 RAWSSVAKS WSFAYMPDLV SRLNRLDLPT LV* It should be noted that this sequence includes a stop codon at position 118. Homology with a predicted ORF from N.εonorrhoeae ORF14 shows 89.8% identity over a 167aa overlap with a predicted ORF (ORF14.ng) from N gonorrhoeae: orf14.pep TAGAAGXXVFVFVTDSQVEVFGΝIQTAVET 30 II III II: I I: I: I :: M M : I II I I orfl4ng GRQFGFFRVGGASFVITAQAGIDDALCDCLTADAAGFAVFAFVADGQMQVFGΝVQPAVET 20E orf14.pep GFFHGISVSSVFGAAAQDSAMASRSASIPVFSATEMRTAAIFPAASRHMPVFCSSDGSRS 90 II I I I II I II I II II II II I I II II I I I II I II I II I I I II I I I I I I I I II I II I I I I I orfl4ng GFFHGISVSSVFGAAAQYSAMASRSASIPVFSATEMRTAAIFPAASRHMPVFCSSDGSRS 268 orf14.pep VLLYTLMHGISPAWISCSTFSTSSICCPLFGAAASTTCSSTSACAVSSSVAEKAEISLCG 150 I II II I II I II II II I II I II II II I I II I II II II II I I I I : II I : I I II I I I I I I I orfl4ng VLLYTLMHGISWAWISCSTFSTSSICCPLFRAAASTTCSSTSACTVSSKVAEKAEISLCG 328 orf14.pep RXLTNPTVSVRIMLHSG 167 I I I I II I II I II IM I orfl4ng RSLTNPTVSVRIMLHAGLMYSRRAWSRVAKSWSFAYMPDLVSRLNRLDLPTLV 382 The complete length ORF14ng nucleotide sequence <SEQ LD 145> is predicted to encode a protein having amino acid sequence <SEQ LD 146>: 1 MEDLQEIGFD VAAVKVGRQR EHHRLHHTQS GNGKADDVLF AFFLVGGFDF 51 LRVIGCGGVA CLPDFQQNVG EADFAWPDD AAAVRAVIEV DADDAVCAQK 101 LLFDQPDAGG AGNAAEHQHC FVRAIMGFHK VGLDFGQWQ ADLVEDFLGR 151 QFGFFRVGGA SFVITAQAGI DDALCDCLTA DAAGFAVFAF VADGQMQVFG 201 NVQPAVETGF FHGISVSSVF GAAAQYSAMA SRSASIPVFS ATEMRTAAIF 251 PAASRHMPVF CSSDGSRSVL LYTLMHGISW AWISCSTFST SSICCPLFRA 301 AASTTCSSTS ACTVSSKVAE KAEISLCGRS LTNPTVSVRI MLHAGLMYSR 351 RAWSRVAKS WSFAYMPDLV SRLNRLDLPT LV* Based on the putative transmembrane domain in the gonococcal protein, it is predicted that the proteins from N. meningitidis and N.gonorrhoeae, and their epitopes, could be useful antigens for vaccines or diagnostics, or for raising antibodies. Example 18 The following partial DNA sequence was identified in N. meningitidis <SEQ ID 147>: 1 .. GGCCATTACT CCGACCGCAC TTGGAAGCCG CGTTTGGNCG GCCGCCGTCT 51 GCCGTATCTG CTTTATGGCA CGCTGATTGC GGTTATTGTG ATGATTTTGA 101 TGCCGAACTC GGGCAGCTTC GGTTTCGGCT ATGCGTCGCT GGCGGCTTTG 151 TCGTTCGGCG CGCTGATGAT TGCGCTGTTA GACGTGTCGT CAAATATGGC 201 GATGCAGCCG TTTAAGATGA TGGTCGGCGA CATGGTCAAC GAGGAGCAGA 251 AAA.NTACGC CTACGGGATT CAAAGTTTCT TAGCAAATAC GGGCGCGGTC 301 GTGGCGGCGA TTCTGCCGTT TGTGTTTGCG TATATCGGTT TGGCGAACAC 351 CGCCGANAAA GGCGTTGTGC CGCAGACCGT GGTCGTGGCG TTTTATGTGG 401 GTGCGGCGTT GCTGGTGATT ACCAGCGCGT TCACGATTTT CAAAGTGAAG 451 GAATACGANC CGGAAACCTA CGCCCGTTAC CACGGCATCG ATGTCGCCGC 501 GAATCAGGAA AAAGCCAACT GGATCGCACT CTTAAAA.CC GCGC .. This corresponds to the amino acid sequence <SEQ ID 148; ORF16>: 1 ..GHYSDRTWKP RLXGRRLPYL LYGTLIAVIV MILMPNSGSF GFGYASLAAL 51 SFGALMIALL DVSSNMAMQP FKMMVGDMVN EEQKXYAYGI QSFLANTGAV 101 VAAILPFVFA YIGLANTAXK GWPQTVWA FYVGAALLVI TSAFTIFKVK 151 EYXPETYARY HGIDVAANQE KANWIALLKX A.. Further work revealed the complete nucleotide sequence <SEQ ID 149>: 1 ATGTCGGAAT ATACGCCTCA AACAGCAAAA CAAGGTTTGC CCGCGCTGGC 51 AAAAAGCACG ATTTGGATGC TCAGTTTCGG CTTTCTCGGC GTTCAGACGG 101 CCTTTACCCT GCAAAGCTCG CAAATGAGCC GCATTTTTCA AACGCTAGGC 151 GCAGACCCGC ACAATTTGGG CTGGTTTTTC ATCCTGCCGC CGCTGGCGGG 201 GATGCTGGTG CAGCCGATTG TCGGCCATTA CTCCGACCGC ACTTGGAAGC 251 CGCGTTTGGG CGGCCGCCGT CTGCCGTATC TGCTTTATGG CACGCTGATT 301 GCGGTTATTG TGATGATTTT GATGCCGAAC TCGGGCAGCT TCGGTTTCGG 351 CTATGCGTCG CTGGCGGCTT TGTCGTTCGG CGCGCTGATG ATTGCGCTGT 401 TAGACGTGTC GTCAAATATG GCGATGCAGC CGTTTAAGAT GATGGTCGGC 451 GACATGGTCA ACGAGGAGCA GAAAGGCTAC GCCTACGGGA TTCAAAGTTT 501 CTTAGCAAAT ACGGGCGCGG TCGTGGCGGC GATTCTGCCG TTTGTGTTTG 551 CGTATATCGG TTTGGCGAAC ACCGCCGAGA AAGGCGTTGT GCCGCAGACC 601 GTGGTCGTGG CGTTTTATGT GGGTGCGGCG TTGCTGGTGA TTACCAGCGC 651 GTTCACGATT TTCAAAGTGA AGGAATACGA TCCGGAAACC TACGCCCGTT 701 ACCACGGCAT CGATGTCGCC GCGAATCAGG AAAAAGCCAA CTGGATCGAA 751 CTCTTGAAAA CCGCGCCTAA GGCGTTTTGG ACGGTTACTT TGGTGCAATT 801 CTTCTGCTGG TTCGCCTTCC AATATATGTG GACTTACTCG GCAGGCGCGA 851 TTGCGGAAAA CGTCTGGCAC ACCACCGATG CGTCTTCCGT AGGTTATCAG 901 GAGGCGGGTA ACTGGTACGG CGTTTTGGCG GCGGTGCAGT CGGTTGCGGC 951 GGTGATTTGT TCGTTTGTAT TGGCGAAAGT GCCGAATAAA TACCATAAGG 1001 CGGGTTATTT CGGCTGTTTG GCTTTGGGCG CGCTCGGCTT TTTCTCCGTT 1051 TTCTTCATCG GCAACCAATA CGCGCTGGTG TTGTCTTATA CCTTAATCGG 1101 CATCGCTTGG GCGGGCATTA TCACTTATCC GCTGACGATT GTGACCAACG 1151 CCTTGTCGGG CAAGCATATG GGCACTTACT TGGGCTTGTT TAACGGCTCT 1201 ATCTGTATGC CTCAAATCGT CGCTTCGCTG TTGAGTTTCG TGCTTTTCCC 1251 TATGCTGGGC GGCTTGCAGG CCACTATGTT CTTGGTAGGG GGCGTCGTCC 1301 TGCTGCTGGG CGCGTTTTCC GTGTTCCTGA TTAAAGAAAC ACACGGCGGG 1351 GTTTGA This corresponds to the amino acid sequence <SEQ ID 150; ORF16-l>: 1 MSEYTPQTAK QGLPALAKST IWMLSFGFLG VQTAFTLQSS QMSRIFQTLG 51 ADPHNLGWFF ILPPLAGMLV QPIVGHYSDR TWKPRLGGRR LPYLLYGTLI 101 AVIVMILMPN SGSFGFGYAS LAALSFGALM IALLDVSSNM AMQPFKMMVG 151 DMVNEEQKGY AYGIQSFLAN TGAWAAILP FVFAYIGLAN TAEKGWPQT 201 WVAFYVGAA LLVITSAFTI FKVKEYDPET YARYHGIDVA ANQEKANWIE 251 LLKTAPKAFW TVTLVQFFCW FAFQYMWTYS AGAIAENVWH TTDASSVGYQ 301 EAGNWYGVLA AVQSVAAVIC SFVLAKVPNK YHKAGYFGCL ALGALGFFSV 351 FFIGNQYALV LSYTLIGIAW AGIITYPLTI VTNALSGKHM GTYLGLFNGS 401 ICMPQIVASL LSFVLFPMLG GLQATMFLVG GWLLLGAFS VFLIKETHGG 451 V* Computer analysis of this amino acid sequence gave the following results: Homology with a predicted ORF from N. meningitidis (strain A) ORF 16 shows 96.7% identity over a 181aa overlap with an ORF (ORF 16a) from strain A of N meningitidis: 10 20 30 orf16. pep GHYSDRTWKPRLXGRRLPYLLYGTLIAVIV I I I I II II II I I I I I II I I I II II I I II I orf16a IFQTLGADPHSLGWFFILPPLAGMLVQPIVGHYSDRTWKPRLGGRRLPYLLYGTLIAVIV 50 60 70 80 90 100 40 50 60 70 80 90 orf16. pep MILMPΝSGSFGFGYASLAALSFGALMIALLDVSSΝMAMQPFKMMVGDMVΝEEQKXYAYGI I I I I I I I I II II I I I I II II I II I II I I I I I I II II II II I II I II II I I II I I II I II orf16a MILMPΝSGSFGFGYASLAALSFGALMIALLDVSSΝMAMQPFKMMVGDMVΝEEQKGYAYGI 110 120 130 140 150 160 100 110 120 130 140 150 orf16. pep QSFLAΝTGAWAAILPFVFAYIGLAΝTAXKGWPQTVWAFYVGAALLVITSAFTIFKVK I I II I II II I II I I II II I I I II I II I I I I I I II I II II I II I I I II I I II I I II II II orflδa QSFLAΝTGAWAAILPFVFAYIGLAΝTAEKGWPQTVWAFYVGAALLVITSAFTIFKVK 170 180 190 200 210 220 160 170 180 orf16. pep EYXPETYARYHGIDVAAΝQEKAΝWIALLKXA II I II II I I I II II I I I I I II II I I IM I orf16a EYΝPETYARYHGIDVAAΝQEKAΝWIELLKTAPKAFWTVTLVQFFCWFAFQYMWTYSAGAI 230 240 250 260 270 280 orf16a AEΝVWHTTDASSVGYQEAGΝWYGVLAAVQSVAAVICSFVLAKVPΝKYHKAGYFGCLALGA 290 300 310 320 330 340 The complete length ORFl 6a nucleotide sequence <SEQ ID 151> is: 1 ATGTCGGAAT ATACGCCTCA AACAGCAAAA CAAGGTTTGC CCGCGCTGGC 51 AAAAAGCACG ATTTGGATGC TCAGTTTCGG CTTTCTCGGC GTTCAGACGG 101 CCTTTACCCT GCAAAGCTCG CAGATGAGCC GCATCTTCCA GACGCTCGGT 151 GCCGATCCGC ACAGCCTCGG CTGGTTCTTT ATCCTGCCGC CGCTGGCGGG 201 GATGCTGGTG CAGCCGATTG TCGGCCATTA CTCCGACCGC ACTTGGAAGC 251 CGCGTTTGGG CGGCCGCCGT CTGCCGTATC TGCTTTATGG CACGCTGATT 301 GCGGTTATTG TGATGATTTT GATGCCGAAC TCGGGCAGCT TCGGTTTCGG 351 CTATGCGTCG CTGGCGGCTT TGTCGTTCGG CGCGCTGATG ATTGCGCTGT 401 TAGACGTGTC GTCAAATATG GCGATGCAGC CGTTTAAGAT GATGGTCGGC 451 GACATGGTCA ACGAGGAGCA GAAAGGCTAC GCCTACGGGA TTCAAAGTTT 501 CTTAGCGAAT ACGGGCGCGG TCGTGGCGGC GATTCTGCCG TTTGTGTTTG 551 CGTATATCGG TTTGGCGAAC ACCGCCGAGA AAGGCGTTGT GCCGCAGACC 601 GTGGTCGTGG CGTTTTATGT GGGTGCGGCG TTGCTGGTGA TTACCAGCGC 651 GTTCACGATT TTCAAAGTGA AGGAATACAA TCCGGAAACC TACGCCCGTT 701 ACCACGGCAT CGATGTCGCC GCGAATCAGG AAAAAGCCAA CTGGATCGAA 751 CTCTTGAAAA CCGCGCCTAA GGCGTTTTGG ACGGTTACTT TGGTGCAATT 801 CTTCTGCTGG TTCGCCTTCC AATATATGTG GACTTACTCG GCAGGCGCGA 851 TTGCGGAAAA CGTCTGGCAC ACCACCGATG CGTCTTCCGT AGGTTATCAG 901 GAGGCGGGTA ACTGGTACGG CGTTTTGGCG GCGGTGCAGT CGGTTGCGGC 951 GGTGATTTGT TCGTTTGTAT TGGCGAAAGT GCCGAATAAA TACCATAAGG 1001 CGGGTTATTT CGGCTGTTTG GCTTTGGGCG CGCTCGGCTT TTTCTCCGTT 1051 TTCTTCATCG GCAACCAATA CGCGCTGGTG TTGTCTTATA CCTTAATCGG 1101 CATCGCTTGG GCGGGCATTA TCACTTATCC GCTGACGATT GTGACCAACG 1151 CCTTGTCGGG CAAGCATATG GGCACTTACT TGGGCCTGTT TAACGGCTCT 1201 ATCTGTATGC CGCAAATCGT CGCTTCGCTG TTGAGTTTCG TGCTTTTCCC 1251 TATGCTGGGC GGCTTGCAGG CCACTATGTT CTTGGTAGGG GGCGTCGTCC 1301 TGCTGCTGGG CGCGTTTTCC GTGTTCCTGA TTAAAGAAAC ACACGGCGGG 1351 GTTTGA This encodes a protein having amino acid sequence <SEQ ID 152>: 1 MSEYTPQTAK QGLPALAKST IWMLSFGFLG VQTAFTLQSS QMSRIFQTLG 51 ADPHSLGWFF ILPPLAGMLV QPIVGHYSDR TWKPRLGGRR LPYLLYGTLI 101 AVIVMILMPN SGSFGFGYAS LAALSFGALM IALLDVSSNM AMQPFKMMVG 151 DMVNEEQKGY AYGIQSFLAN TGAWAAILP FVFAYIGLAN TAEKGWPQT 201 VWAFYVGAA LLVITSAFTI FKVKEYNPET YARYHGIDVA ANQEKANWIE 251 LLKTAPKAFW TVTLVQFFCW FAFQYMWTYS AGAIAENVWH TTDASSVGYQ 301 EAGNWYGVLA AVQSVAAVIC SFVLAKVPNK YHKAGYFGCL ALGALGFFSV 351 FFIGNQYA-LV LSYTLIGIAW AGIITYPLTI VTNALSGKHM GTYLGLFNGS 401 ICMPQIVASL LSFVLFPMLG GLQATMFLVG GVVLLLGAFS VFLIKETHGG 451 V* ORF16a and ORF16-1 show 99.6% identity in 451 aa overlap: 10 20 30 40 50 60 orf16a. pep MSEYTPQTAKQGLPALAKSTIWMLSFGFLGVQTAFTLQSSQMSRIFQTLGADPHSLGWFF I II II I I II I II I II I II II I I I II I I II II II I II II I I I I I II I I I II II I M II I II orf16-1 MSEYTPQTAKQGLPALAKSTIWMLSFGFLGVQTAFTLQSSQMSRIFQTLGADPHNLGWFF 10 20 30 40 50 60 70 80 90 100 110 120 orf16a. pep ILPPLAGMLVQPIVGHYSDRTWKPRLGGRRLPYLLYGTLIAVIVMILMPNSGSFGFGYAS I I II II I I I I II I I II I I I II I I I II II II II II I II II I II II I I II II I II I I I II I I orf16-1 ILPPLAGMLVQPIVGHYSDRTWKPRLGGRRLPYLLYGTLIAVIVMILMPNSGSFGFGYAS 70 80 90 100 110 120 130 140 150 160 170 180 orf16a . pep LAALSFGALMIALLDVSSNMAMQPFKMMVGDMVNEEQKGYAYGIQSFLANTGAWAAILP I I II II I I I II II I II I II I II I I I II I I II I I I I I II II I I I I II I II I II I I II II I I orf16-1 LAALSFGALMIALLDVSSNMAMQPFKMMVGDMVNEEQKGYAYGIQSFLANTGAWAAILP 130 140 150 160 170 180 190 200 210 220 230 240 orf16a . pep FVFAYIGLANTAEKGWPQTVWAFYVGAALLVITSAFTIFKVKEYNPETYARYHGIDVA I II II I I I II I II I II I II I II I I I I I I I II I I I II II II I I I II I: II I II II I II I I I orf16-1 FVFAYIGLANTAEKGVVPQTVWAFYVGAALLVITSAFTIFKVKEYDPETYARYHGIDVA 190 200 210 220 230 240 250 260 270 280 290 300 orf16a . pep ANQEKANWIELLKTAPKAFWTVTLVQFFCWFAFQYMWTYSAGAIAENVWHTTDASSVGYQ II I I I I I I II II I I II I II I II I I I II I I II II I II II II I I I II II I II I II I I I II I I orf16-1 ANQEKANWIELLKTAPKAFWTVTLVQFFCWFAFQYMWTYSAGAIAENVWHTTDASSVGYQ 250 260 270 280 290 300 310 320 330 340 350 360 orf16a . pep EAGNWYGVLAAVQSVAAVICSFVLAKVPNKYHKAGYFGCLALGALGFFSVFFIGNQYALV I I I I I II I I I I II II I I I I II I I I II I I I I I II I I I II I II I I II I I II I II I I I II I I I orf16-1 EAGNWYGVLAAVQSVAAVICSFVLAKVPNKYHKAGYFGCLALGALGFFSVFFIGNQYALV 310 320 330 340 350 360 370 380 390 400 410 420 orf16a. pep LSYTLIGIAWAGIITYPLTIVTNALSGKHMGTYLGLFNGSICMPQIVASLLSFVLFPMLG I II I II I I I I I I II I I II I II I I I I I II II I II II I I II II I I II I I M I II I I I I II I I orf16-1 LSYTLIGIAWAGIITYPLTIVTNALSGKHMGTYLGLFNGSICMPQIVASLLSFVLFPMLG 370 380 390 400 410 420 430 440 450 orf16a . pep GLQATMFLVGGWLLLGAFSVFLIKETHGGVX I I II I I I I I M I II I I I I I I II I I M I I I II I orf16-1 GLQATMFLVGGWLLLGAFSVFLIKETHGGVX 430 440 450 Homology with a predicted ORF from Nsonorrhoeae ORF 16 shows 93.9% identity over a 181aa overlap with a predicted ORF (ORFlό.ng) from N. gonorrhoeae: orf16. pep GHYSDRTWKPRLXGRRLPYLLYGTLIAVIV 30 I : II II II I I II I I I II I I II II I I I I I I orf16ng HFSΝARRRPAQFGLVFHPAAAGGDAGSADSGYYSDRTWKPRLGGRRLPYLLYGTLIAVIV 131 orf16. pep MILMPΝSGSFGFGYASLAALSFGALMIALLDVSSΝMAMQPFKMMVGDMVΝEEQKXYAYGI 90 II II II II II II II I II II I I I I I II I I I I I II I I II I I I I II I I I I II I I II I I I I I I orf16ng MILMPΝSGSFGFGYASLAALSFGALMIALLDVSSΝMAMQPFKMMVGDMVΝEEQKSYAYGI 191 orf16. pep QSFLAΝTGAWAAILPFVFAYIGLAΝTAXKGWPQTWVAFYVGAALLVITSAFTIFKVK 150 II II I II I II I I I I I II II II I II I I I I II I I I I II I II II II I I I : I I II II I III orf16ng QSFLAΝTDAWAAILPFVFAYIGLAΝTAEKGWPQTVWAFYVGAALLIITSAFTISKVK 251 orf16.pep EYXPETYARYHGIDVAAΝQEKAΝWIALLKXA 181 II II I I I II II II II I I I II I I M II I : I orf16ng EYDPETYARYHGIDVAAΝQEKAΝWFELLKTAPKVFWTVTPVQFFCWFAFRYMWTYSAGAI 311 The complete length ORF16ng nucleotide sequence <SEQ ID 153> is: 1 ATGATAGGGG ATCGCCGCGC CGGCAACCAT TTCGGATTTT CCAAAGCAAA 51 TACTTTTCAA ATCAAAAAAA AGGATTTACT TTATGTCGGA ATATACGCCT 101 CAAACAGCAA AACAAGGTTT GCCCGCGCCG GCAAAAAGCA CGATTTGGAT 151 GTTGAGCTTC GGCTATCTCG GCGTTCAGAC GGCCTTTACC CTGCAAAGCT 201 CGCAGATGAG CCGCATTTTT CAAACGCTAG GCGCAGACCC GCACAATTTG 251 GGCTGGTTTT TCATCCTGCC GCCGCTGGCG GGGATGCTGG TTCAGCCGAT 301 AGTGGCTACT ACTCAGACCG CACTTGGAAG CCGCGCTTGG GCGGCCGCCG 351 CCTGCCGTAT CTGCTTTACG GCACGCTGAT TGCGGTCATC GTGATGATTT 401 TGATGCCGAA CTCGGGCAGC TTCGGTTTCG GCTATGCGTC GCTGGCGGCC 451 TTGTCGTTCG GCGCGCTGAT GATTGCGCTG TTGGACGTGT CGTCGAATAT 501 GGCGATGCAG CCGTTTAAGA TGATGGTCGG CGATATGGTC AACGAGGAGC 551 AGAAAAGCTA CGCCTACGGG ATTCAAAGTT TCTTAGCGAA TACGGACGCG 601 GTTGTGGCAG CGATTCTGCC GTTTGTGTTC GCGTATATCG GTTTGGCGAA 651 CACTGCCGAG AAAGGCGTTG TGCCACAAAC CGTGGTCGTA GCATTCTATG 701 TGGGTGCGGC GTTACTGATT ATTACCAGTG CGTTCACAAT CTCCAAAGTC 751 AAAGAATACG ACCCGGAAAC CTACGCCCGT TACCACGGCA TCGATGTCGC 801 CGCGAATCAG GAAAAAGCCA ACTGGTTCGA ACTCTTAAAA ACCGCGCCTA 851 AAGTGTTTTG GACGGTTACT CCGGTACAGT TTTTCTGCTG GTTCGCCTTC 901 CGGTATATGT GGACTTACTC GGCAGGCGCG ATTGCAGAAA ACGTCTGGCA 951 CACTACCGAT GCGTCTTCCG TAGGCCATCA GGAGGCGGGC AACCGGTACG 1001 GCGTTTTGGC GGCGGTGTAG This encodes a protein having amino acid sequence <SEQ LD 154>: 1 MIGDRRAGΝH FGFSKAΝTFQ IKKKDLLYVG IYASΝSKTRF ARAGKKHDLD 51 VELRLSRRSD GLYPAKLADE PHFSΝARRRP AQFGLVFHPA AAGGDAGSAD 101 SGYYSDRTWK PRLGGRRLPY LLYGTLIAVI VMILMPΝSGS FGFGYASLAA 151 LSFGALMIAL LDVSSΝMAMQ PFKMMVGDMV ΝEEQKSYAYG IQSFLAΝTDA 201 WAAILPFVF AYIGLAΝTAE KGWPQTWV AFYVGAALLI ITSAFTISKV 251 KEYDPETYAR YHGIDVAAΝQ EKAΝWFELLK TAPKVFWTVT PVQFFCWFAF 301 RYMWTYSAGA IAEΝVWHTTD ASSVGHQEAG ΝRYGVLAAV* ORFlόng and ORF16-1 show 89.3% identity in 261 aa overlap: 30 40 50 60 70 80 orf16-1. pep MLSFGFLGVQTAFTLQSSQMSRIFQTLGADPHΝLGWFFILPPLAGMLVQPI-VGHYSDRT I :: I I I II : M I I I II orfl6ng DVELRLSRRSDGLYPAKLADEPHFSΝARRRPAQFGLVF-HPAAAGGDAGSADSGYYSDRT 50 60 70 80 90 100 90 100 110 120 130 140 orf16-1. pep WKPRLGGRRLPYLLYGTLIAVIVMILMPΝSGSFGFGYASLAALSFGALMIALLDVSSΝMA I I II I I II I II II I I I I I I I I I II II I I I I II II II I I I I I I II I II I I II II II II I I I orflδng WKPRLGGRRLPYLLYGTLIAVIVMILMPΝSGSFGFGYASLAALSFGALMIALLDVSSΝMA 110 120 130 140 150 160 150 160 170 180 190 200 orf16-1.pep MQPFKMMVGDMVNEEQKGYAYGIQSFLANTGAWAAILPFVFAYIGLANTAEKGWPQTV I II I I I II II II I I II I : I II I I I II II II I I II I I II I I I II I I I I II II I I I I I I II orflδng MQPFKMMVGDMVNEEQKSYAYGIQSFLANTDAWAAILPFVFAYIGLANTAEKGWPQTV 170 180 190 200 210 220 210 220 230 240 250 260 orf16-1. pep WAFYVGAALLVITSAFTIFKVKEYDPETYARYHGIDVAANQEKANWIELLKTAPKAFWT I II II I II II I : I I II I I I II II I I I I I I I II I I II I II I I I II I I : II I II I I I : I I I orflδng WAFYVGAALLIITSAFTISKVKEYDPETYARYHGIDVAANQEKANWFELLKTAPKVFWT 230 240 250 260 270 280 270 280 290 300 310 320 orf16-1.pep VTLVQFFCWFAFQYMWTYSAGAIAENVWHTTDASSVGYQEAGNWYGVLAAVQSVAAVICS II I II II I I II : I I II II I I II II I I I I II I II II I : I I I I I I I I I I I I orf16ng VTPVQFFCWFAFRYMWTYSAGAIAENVWHTTDASSVGHQEAGNRYGVLAAVX 290 300 310 320 330 340 Based on this analysis, including the presence of several putative transmembrane domains in the gonococcal protein, it is predicted that the proteins from N. meningitidis and N.gonorrhoeae, and their epitopes, could be useful antigens for vaccines or diagnostics, or for raising antibodies. Example 19 The following partial DNA sequence was identified in N meningitidis <SEQ ID 155>: 1 ATGTTGTTCC GTAAAACGAC CGCCGCCGTT TTGGCGCATA CCTTGATGCT 51 GAACGGCTGT ACGTTGATGT TGTGGGGAAT GAACAACCCG GTCAGCGAAA 101 CAATCACCCG ΝAAACACGTT GΝCAAAGACC AAATCCGΝGΝ CTTCGGTGTG 151 GTTGCCGAAG ACAATGCCCA ATTGGAAAAG GGCAGCCTGG TGATGATGGG 201 CGGAAAATAC TGGTTCGTCG TCAATCCCGA AGATTCGGCG AA.ΝTGACGG 251 GΝATTTTGAΝ GGCAGGGCTG GACAAACCCT TCCAAATAGT TΝAGGATACC 301 CCGAGCTATG C.TGCCACCA AGCCCTGCCG GTCAAACTCG GATCGΝCTGG 351 CAGCCAGAAT ... This corresponds to the amino acid sequence <SEQ ID 156; ORF28>: 1 MLFRKTTAAV LAHTLMLΝGC TLMLWGMΝΝP VSETITRKHV XKDQIRXFGV 51 VAEDΝAQLEK GSLVMMGGKY WFWΝPEDSA XXTGILXAGL DKPFQIVXDT 101 PSYXCHQALP VKLGSXGSQ ... Further work revealed the complete nucleotide sequence <SEQ LD 157>: 1 ATGTTGTTCC GTAAAACGAC CGCCGCCGTT TTGGCGGCAA CCTTGATGCT 51 GAACGGCTGT ACGTTGATGT TGTGGGGAAT GAACAACCCG GTCAGCGAAA 101 CAATCACCCG CAAACACGTT GACAAAGACC AAATCCGCGC CTTCGGTGTG 151 GTTGCCGAAG ACAATGCCCA ATTGGAAAAG GGCAGCCTGG TGATGATGGG 201 CGGAAAATAC TGGTTCGTCG TCAATCCCGA AGATTCGGCG AAGCTGACGG 251 GCATTTTGAA GGCAGGGCTG GACAAACCCT TCCAAATAGT TGAGGATACC 301 CCGAGCTATG CTCGCCACCA AGCCCTGCCG GTCAAACTCG AATCGCCTGG 351 CAGCCAGAAT TTCAGTACCG AAGGCCTTTG CCTGCGCTAC GATACCGACA 401 AGCCTGCCGA CATCGCCAAG CTGAAACAGC TCGGGTTTGA AGCGGTCAAA 451 CTCGACAATC GGACCATTTA CACGCGCTGC GTATCCGCCA AAGGCAAATA 501 CTACGCCACA CCGCAAAAAC TGAACGCCGA TTACCATTTT GAGCAAAGTG 551 TGCCTGCCGA TATTTATTAC ACGGTTACTG AAGAACATAC CGACAAATCC 601 AAGCTGTTTG CAAATATCTT ATATACGCCC CCCTTTTTGA TACTGGATGC 651 GGCGGGCGCG GTACTGGCCT TGCCTGCGGC GGCTCTGGGT GCGGTCGTGG 701 ATGCCGCCCG CAAATGA This corresponds to the amino acid sequence <SEQ ID 158; ORF28-l>: 1 MLFRKTTAAV LAATLMLΝGC TLMLWGMΝΝP VSETITRKHV DKDQIRAFGV 51 VAEDΝAQLEK GSLVMMGGKY WFWΝPEDSA KLTGILKAGL DKPFQIVEDT 101 PSYARHQALP VKLESPGSQΝ FSTEGLCLRY DTDKPADIAK LKQLGFEAVK 151 LDΝRTIYTRC VSAKGKYYAT PQKLΝADYHF EQSVPADIYY TVTEEHTDKS 201 KLFANILYTP PFLILDAAGA VLALPAAALG AWDAARK* Computer analysis of this amino acid sequence gave the following results: Homology with a predicted ORF from N. meningitidis (strain A) ORF28 shows 79.2% identity over a 120aa overlap with an ORF (ORF28a) from strain A of N. meningitidis: 10 20 30 40 50 60 orf28.pep MLFRKTTAAVLAHTLMLNGCTLMLWGMNNPVSETITRKHVXKDQIRXFGWAEDNAQLEK I II I II II I II I II I I II I I : I : I II I: I III Mill II II I I I I II I II I I I I I orf28a MLFRKTTAAVLAATLMLNGCTVMMWGMNSPFSETTARKHVDKDQIRAFGWAEDNAQLEK 10 20 30 40 50 60 70 80 90 100 110 120 orf28.pep GSLVMMGGKYWFWNPEDSAXXTGILXAGLDKPFQIVXDTPSYXCHQALPVKLGSXGSQN I I I II I I I II II I I I II II I I I II I II II I Ml :| : : II II II I I Ml I orf28a GSLVMMGGKYWFWNPEDSAKLTGILKAGLDKQFQMVEPNPRFA-YQALPVKLESPASQN 70 80 90 100 110 orf28a FSTEGLCLRYDTDRPADIAKLKQLEFEAVELDNRTIYTRCVSAKGKYYATPQKLNADYHF 120 130 140 150 160 170 The complete length ORF28a nucleotide sequence <SEQ ID 159> is: 1 ATGTTGTTCC GTAAAACGAC CGCCGCCGTT TTGGCGGCAA CCTTGATGTT 51 GAACGGCTGT ACGGTAATGA TGTGGGGTAT GAACAGCCCG TTCAGCGAAA 101 CGACCGCCCG CAAACACGTT GACAAGGACC AAATCCGCGC CTTCGGTGTG 151 GTTGCCGAAG ACAATGCCCA ATTGGAAAAG GGCAGCCTGG TGATGATGGG 201 CGGGAAATAC TGGTTCGTCG TCAATCCTGA AGATTCGGCG AAGCTGACGG 251 GCATTTTGAA GGCCGGGTTG GACAAGCAGT TTCAAATGGT TGAGCCCAAC 301 CCGCGCTTTG CCTACCAAGC CCTGCCGGTC AAACTCGAAT CGCCCGCCAG 351 CCAGAATTTC AGTACCGAAG GCCTTTGCCT GCGCTACGAT ACCGACAGAC 401 CTGCCGACAT CGCCAAGCTG AAACAGCTTG AGTTTGAAGC GGTCGAACTC 451 GACAATCGGA CCATTTACAC GCGCTGCGTC TCCGCCAAAG GCAAATACTA 501 CGCCACACCG CAAAAACTGA ACGCCGATTA TCATTTTGAG CAAAGTGTGC 551 CTGCCGATAT TTATTACACG GTTACGAAAA AACATACCGA CAAATCCAAG 601 TTGTTTGAAA ATATTGCATA TACGCCCACC ACGTTGATAC TGGATGCGGT 651 GGGCGCGGTG CTGGCCTTGC CTGTCGCGGC GTTGATTGCA GCCACGAATT 701 CCTCAGACAA ATGA This encodes a protein having amino acid sequence <SEQ ID 160>: 1 MLFRKTTAAV LAATLMLNGC TVMMWGMNSP FSETTARKHV DKDQIRAFGV 51 VAEDNAQLEK GSLVMMGGKY WFWNPEDSA KLTGILKAGL DKQFQMVEPN 101 PRFAYQALPV KLESPASQNF STEGLCLRYD TDRPADIAKL KQLEFEAVEL 151 DNRTIYTRCV SAKGKYYATP QKLNADYHFE QSVPADIYYT VTKKHTDKSK 201 LFENIAYTPT TLILDAVGAV LALPVAALIA ATNSSDK* ORF28a and ORF28-1 show 86.1% identity in 238 aa overlap: 10 20 30 40 50 60 orf28a.pep MLFRKTTAAVLAATLMLNGCTVMMWGMNSPFSETTARKHVDKDQIRAFGWAEDNAQLEK I M I I I M I II I I II II I II I : I : II I I : I III M II I II II II I M I II I I I I I II I orf28-1 MLFRKTTAAVLAATLMLNGCTLMLWGMNNPVSETITRKHVDKDQIRAFGWAEDNAQLEK 10 20 30 40 50 60 70 80 90 100 110 119 orf28a.pep GSLVMMGGKYWFWNPEDSAKLTGILKAGLDKQFQMVEPNPRFA-YQALPVKLESPASQN I I I I I I II II I I I I I I I I II II I I II I II II I I M : I : I : II II II I I IM II I orf28-1 GSLVMMGGKYWFWNPEDSAKLTGILKAGLDKPFQIVEDTPSYARHQALPVKLESPGSQN 70 80 90 100 110 120 120 130 140 150 160 170 179 orf28 .pep FSTEGLCLRYDTDRPADIAKLKQLEFEAVELDNRTIYTRCVSAKGKYYATPQKLNADYHF II II I II II I II I : I I I II I I I I I I II I: I II II I I I II I II I II I I II II I I II I II I orf28-1 FSTEGLCLRYDTDKPADIAKLKQLGFEAVKLDNRTIYTRCVSAKGKYYATPQKLNADYHF 130 140 150 160 170 180 180 190 200 210 220 230 orf28a . pep EQSVPADIYYTVTKKHTDKSKLFENIAYTPTTLILDAVGAVLALPVAALIAATNSSDKX II II II II II I II :: II I I II I I II III II I II : II II II I : II I I ::::: II orf28-l EQSVPADIYYTVTEEHTDKSKLFANILYTPPFLILDAAGAVLALPAAALGAWDAARKX 190 200 210 220 230 Homology with a predicted ORF from N.sonorrhoeae ORF28 shows 84.2% identity over a 120aa overlap with a predicted ORF (ORF28.ng) from N. gonorrhoeae: orf28.pep MLFRKTTAAVLAHTLMLNGCTLMLWGMNNPVSETITRKHVXKDQIRXFGWAEDNAQLEK 60 II I II I I I II II II : I II I I : II I I I I II I : II II II I II II I I I I II I I II I I II orf28ng MLFRKTTAAVLAATLILNGCTMMLRGMNNPVSQTITRKHVDKDQIRAFGWAEDNAQLEK 60 orf28.pep GSLVMMGGKYWFWNPEDSAXXTGILXAGLDKPFQIVXDTPSYXCHQALPVKLGSXGSQN 120 II I II I I I II II : II II II I MM II II II I I I I II II I I II II II : : I I II orf28ng GSLVMMGGKYWFAVNPEDSAKLTGLLKAGLDKPFQIVEDTPSYARHQALPVKFEAPGSQN 120 The complete length ORF28ng nucleotide sequence <SEQ ID 161> is 1 ATGTTGTTCC GTAAAACGAC CGCCGCCGTT TTGGCGGCAA CCTTGATACT 51 GAACGGCTGT ACGATGATGT TGCGGGGGAT GAACAACCCG GTCAGCCAAA 101 CAATCACCCG CAAACACGTT GACAAAGACC AAATCCGCGC CTTCGGTGTG 151 GTTGCCGAAG ACAATGCCCA ATTGGAAAAG GGCAGCCTGG TGATGATGGG 201 CGGGAAATAC TGGTTCGCCG TCAATCCCGA AGATTCGGCG AAGCTGACGG 251 GCCTTTTGAA GGCCGGGTTG GACAAGCCCT TCCAAATAGT TGAGGATACC 301 CCGAGCTATG CCCGCCACCA AGCCCTGCCG GTCAAATTCG AAGCGCCCGG 351 CAGCCAGAAT TTCAGTACCG GAGGTCTTTG CCTGCGCTAT GATACCGGCA 401 GACCTGACGA CATCGCCAAG CTGAAACAGC TTGAGTTTAA AGCGGTCAAA 451 CTCGACAATC GGACCATTTA CACGCGCTGC GTATCCGCCA AAGGCAAATA 501 CTACGCCACG CCGCAAAAAC TGAACGCCGA TTATCATTTT GAGCAAAGTG 551 TGCCCGCCGA TATTTATTAT ACGGTTACTG AAAAACATAC CGACAAATCC 601 AAGCTGTTTG GAAATATCTT ATATACGCCC CCCTTGTTGA TATTGGATGC 651 GGCGGCCGCG GTGCTGGTCT TGCCTATGGC TCTGATTGCA GCCGCGAATT 701 CCTCAGACAA ATGA This encodes a protein having amino acid sequence <SEQ ID 162>: 1 MLFRKTTAAV LAATLILNGC TMMLRGMNNP VSQTITRKHV DKDQIRAFGV 51 VAEDNAQLEK GSLVMMGGKY WFAVNPEDSA KLTGLLKAGL DKPFQIVEDT 101 PSYARHQALP VKFEAPGSQN FSTGGLCLRY DTGRPDDIAK LKQLEFKAVK 151 LDNRTIYTRC VSAKGKYYAT PQKLNADYHF EQSVPADIYY TVTEKHTDKS 201 KLFGNILYTP PLLILDAAAA VLVLPMALIA AANSSDK* ORF28ng and ORF28-1 share 90.0% identity in 231 aa overlap: 10 20 30 40 50 60 orf28-1. pep MLFRKTTAAVLAATLMLNGCTLMLWGMNNPVSETITRKHVDKDQIRAFGWAEDNAQLEK I I I II I I II II I I I I: I I II I: I I I II II II : II II I I II II I II I II I II I I I I I I II orf28ng MLFRKTTAAVLAATLILNGCTMMLRGMNNPVSQTITRKHVDKDQIRAFGWAEDNAQLEK 10 20 30 40 50 60 70 80 90 100 110 120 orf28-1. pep GSLVMMGGKYWFWNPEDSAKLTGILKAGLDKPFQIVEDTPSYARHQALPVKLESPGSQN I I I II I I II II II I I I I M I II: I I I I II II I II II II II I I I II I I II I: I I II I orf28ng GSLVMMGGKYWFAVNPEDSAKLTGLLKAGLDKPFQIVEDTPSYARHQALPVKFEAPGSQN 70 80 90 100 110 120 130 140 150 160 170 180 orf28-1. pep FSTEGLCLRYDTDKPADIAKLKQLGFEAVKLDNRTIYTRCVSAKGKYYATPQKLNADYHF II I I I I II I I I : I II I I II II I : I I II I I I II II II II II I II II II I I I I I I I II orf28ng FSTGGLCLRYDTGRPDDIAKLKQLEFKAVKLDNRTIYTRCVSAKGKYYATPQKLNADYHF 130 140 150 160 170 180 190 200 210 220 230 239 orf28-1. pep EQSVPADIYYTVTEEHTDKSKLFANILYTPPFLILDAAGAVLALPAAALGAWDAARKX II I II I II I I I II M I I II I II MM I I I IM I I II I I: I IMM I :: I: orf28ng EQSVPADIYYTVTEKHTDKSKLFGNILYTPPLLILDAAAAVLVLPMALIAAANSSDKX 190 200 210 220 230 Based on this analysis, including the presence of a putative transmembrane domain in the gonococcal protein, it was predicted that the proteins from N. meningitidis .and N.gonorrhoeae, and their epitopes, could be useful antigens for vaccines or diagnostics, or for raising antibodies. ORF28-1 (24kDa) was cloned in pET and pGex vectors and expressed in E.coli, as described above. The products of protein expression and purification were analyzed by SDS-PAGE. Figure 6 A shows the results of affinity purification of the GST-fusion protein, and Figure 6B shows the results of expression of the His-fusion in E.coli. Purified GST-fusion protein was used to immunise mice, whose sera were used for ELISA, which gave a positive result. These experiments confirm that ORF28-1 is a surface-exposed protein, and that it may be a useful immunogen. Example 20 The following partial DNA sequence was identified in N. meningitidis <SEQ ID 163>: 1 .. GTCAGTCCTG TACTGCCTAT TACACACGAA CGGACAGGGT TTGAAGGTGT 51 TATCGGTTAT GAAACCCATT TTTCAGGGCA CGGACATGAA GTACACAGTC 101 CGTTCGATCA TCATGATTCA AAAAGCACTT CTGATTTCAG CGGCGGTGTA 151 GACGGCGGTT TTACTGTTTA CCAACTTCAT CGAACATGGT CGGAAATCCA 201 TCCGGAGGAT GAATATGACG GGCCGCAAGC AGCG.ATTAT CCGCCCCCCG 251 GAGGAGCAAG GGATATATAC AGCTATTATG TCAAAGGAAC TTCAACAAAA 301 ACAAAGACTA GTATTGTCCC TCAAGCCCCA TTTTCAGACC GTTGGCTAGA 351 AGAAAATGCC GGTGCCGCCT CTGGT.. This corresponds to the amino acid sequence <SEQ ID 164; ORF29>: 1 ..VSPVLPITHE RTGFEGVIGY ETHFSGHGHE VHSPFDHHDS KSTSDFSGGV 51 DGGFTVYQLH RTWSEIHPED EYDGPQAAXY PPPGGARDIY SYYVKGTSTK 101 TKTSIVPQAP FSDRWLEENA GAASG.. Further work revealed the complete nucleotide sequence <SEQ LD 165>: 1 ATGAATTTGC CTATTCAAAA ATTCATGATG CTGTTTGCAG CAGCAATATC 51 GTTGCTGCAA ATCCCCATTA GTCATGCGAA CGGTTTGGAT GCCCGTTTGC 101 GCGATGATAT GCAGGCAAAA CACTACGAAC CGGGTGGTAA ATACCATCTG 151 TTTGGTAATG CTCGCGGCAG TGTTAAAAAG CGGGTTTACG CCGTCCAGAC 201 ATTTGATGCA ACTGCGGTCA GTCCTGTACT GCCTATTACA CACGAACGGA 251 CAGGGTTTGA AGGTGTTATC GGTTATGAAA CCCATTTTTC AGGGCACGGA 301 CATGAAGTAC ACAGTCCGTT CGATCATCAT GATTCAAAAA GCACTTCTGA 351 TTTCAGCGGC GGTGTAGACG GCGGTTTTAC TGTTTACCAA CTTCATCGAA 401 CAGGGTCGGA AATCCATCCG GAGGATGGAT ATGACGGGCC GCAAGGCAGC 451 GATTATCCGC CCCCCGGAGG AGCAAGGGAT ATATACAGCT ATTATGTCAA 501 AGGAACTTCA ACAAAAACAA AGACTAATAT TGTCCCTCAA GCCCCATTTT 551 CAGACCGTTG GCTAAAAGAA AATGCCGGTG CCGCCTCTGG TTTTTTCAGC 601 CGTGCGGATG AAGCAGGAAA ACTGATATGG GAAAGCGACC CCAATAAAAA 651 TTGGTGGGCT AACCGTATGG ATGATGTTCG CGGCATCGTC CAAGGTGCGG 701 TTAATCCTTT TTTAATGGGT TTTCAAGGAG TAGGGATTGG GGCAATTACA 751 GACAGTGCAG TAAGCCCGGT CACAGATACA GCCGCGCAGC AGACTCTACA 801 AGGTATTAAT GATTTAGGAA AATTAAGTCC GGAAGCACAA CTTGCTGCCG 851 CGAGCCTATT ACAGGACAGT GCTTTTGCGG TAAAAGACGG TATCAACTCT 901 GCCAAACAAT GGGCTGATGC CCATCCAAAT ATAACAGCTA CTGCCCAAAC 951 TGCCCTTTCC GCAGCAGAGG CCGCAGGTAC GGTTTGGAGA GGTAAAAAAG 1001 TAGAACTTAA CCCGACTAAA TGGGATTGGG TTAAAAATAC CGGTTATAAA 1051 AAACCTGCTG CCCGCCATAT GCAGACTTTA GATGGGGAGA TGGCAGGTGG 1101 GAATAAACCT ATTAAATCTT TACCAAACAG TGCCGCTGAA AAAAGAAAAC 1151 AAAATTTTGA GAAGTTTAAT AGTAACTGGA GTTCAGCAAG TTTTGATTCA 1201 GTGCACAAAA CACTAACTCC CAATGCACCT GGTATTTTAA GTCCTGATAA 1251 AGTTAAAACT CGATACACTA GTTTAGATGG AAAAATTACA ATTATAAAAG 1301 ATAACGAAAA CAACTATTTT AGAATCCATG ATAATTCACG AAAACAGTAT 1351 CTTGATTCAA ATGGTAATGC TGTGAAAACC GGTAATTTAC AAGGTAAGCA 1401 AGCAAAAGAT TATTTACAAC AACAAACTCA TATCAGGAAC TTAGACAAAT 1451 GA This corresponds to the amino acid sequence <SEQ ID 166; ORF29-l>: 1 MNLPIQKFMM LFAAAISLLQ IPISHANGLD ARLRDDMQAK HYEPGGKYHL 51 FGNARGSVKK RVYAVQTFDA TAVSPVLPIT HERTGFEGVI GYETHFSGHG 101 HEVHSPFDHH DSKSTSDFSG GVDGGFTVYQ LHRTGSEIHP EDGYDGPQGS 151 DYPPPGGARD IYSYYVKGTS TKTKTNIVPQ APFSDRWLKE NAGAASGFFS 201 RADEAGKLIW ESDPNKNWWA NRMDDVRGIV QGAVNPFLMG FQGVGIGAIT 251 DSAVSPVTDT AAQQTLQGIN DLGKLSPEAQ LAAASLLQDS AFAVKDGINS 301 AKQWADAHPN ITATAQTALS AAEAAGTVWR GKKVELNPTK WDWVKNTGYK 351 KPAARHMQTL DGEMAGGNKP IKSLPNSAAE KRKQNFEKFN SNWSSASFDS 401 VHKTLTPNAP GILSPDKVKT RYTSLDGKIT IIKDNENNYF RIHDNSRKQY 451 LDSNGNAVKT GNLQGKQAKD YLQQQTHIRN LDK* Computer analysis of this amino acid sequence gave the following results: Homology with a predicted ORF from N.meninsitidis (strain Al ORF29 shows 88.0% identity over a 125aa overlap with an ORF (ORF29a) from strain A of N. meningitidis: 10 20 30 orf29. pep VSPVLPITHERTGFEGVIGYETHFSGHGHE I : I: II I II II II II I : II I I I I II II II I orf29a EPGGKYHLFGΝARGSVKΝRVYAVQTFDATAVGPILPITHERTGFEGIIGYETHFSGHGHE 50 60 70 80 90 100 40 50 60 70 80 90 orf29. pep VHSPFDHHDSKSTSDFSGGVDGGFTVYQLHRTWSEIHPEDEYDGPQAAXYPPPGGARDIY M II I I : II I II II II II II II II II I I II I I I II I II I Mill:: II II I II II II orf29a VHSPFDΝHDSKSTSDFSGGVDGGFTVYQLHRTGSEIHPEDGYDGPQGSDYPPPGGARDIY 110 120 130 140 150 160 100 110 120 orf29.pep SYYVKGTSTKTKTSIVPQAPFSDRWLEEΝAGAASG I I I II II II I :: II M I I II II II : I II I II I I orf29a XXYVKGTSTKTKSΝIVPRAPFSDRWLKEΝAGAASGFFSRADEAGKLIWESDPΝKΝWWAΝR 170 180 190 200 210 220 orf29a MDDIRGIVQGAVΝPFLMGFQGVGIGAITDSAVSPVTDTAAQQTLQGXΝHLGXLSPEAQLA 230 240 250 260 270 280 The complete length ORF29a nucleotide sequence <SEQ ID 167> is: 1 ATGAATTΝGC CTATTCAAAA ATTCATGATG CTGTTTGCAG CAGCAATATC 51 GTΝGCTGCAA ATCCCΝATTA GTCATGCGAA CGGTTTGGAT GCCCGTTTGC 101 GCGATGATAT GCAGGCAAAA CACTACGAAC CGGGTGGTAA ATACCATCTG 151 TTTGGTAATG CTCGCGGCAG TGTTAAAAAT CGGGTTTACG CCGTCCAAAC 201 ATTTGATGCA ACTGCGGTCG GCCCCATACT GCCTATTACA CACGAACGGA 251 CAGGATTTGA AGGCATTATC GGTTATGAAA CCCATTTTTC AGGACATGGA 301 CATGAAGTAC ACAGTCCGTT CGATAATCAT GATTCAAAAA GCACTTCTGA 351 TTTCAGCGGC GGCGTAGACG GTGGTTTTAC CGTTTACCAA CTTCATCGGA 401 CAGGGTCGGA AATCCATCCG GAGGATGGAT ATGACGGGCC GCAAGGCAGC 451 GATTATCCGC CCCCCGGAGG AGCAAGGGAT ATATACAΝΝT AΝTATGTCAA 501 AGGAACTTCA ACAAAAACAA AGAGTAATAT TGTTCCCCGA GCCCCATTTT 551 CAGACCGCTG GCTAAAAGAA AATGCCGGTG CCGCCTCTGG TTTTTTCAGC 601 CGTGCTGATG AAGCAGGAAA ACTGATATGG GAAAGCGACC CCAATAAAAA 651 TTGGTGGGCT AACCGTATGG ATGATATTCG CGGCATCGTC CAAGGTGCGG 701 TTAATCCTTT TTTAATGGGT TTTCAAGGAG TAGGGATTGG GGCAATTACA 751 GACAGTGCAG TAAGCCCGGT CACAGATACA GCCGCGCAGC AGACTCTACA 801 AGGTATΝAAT CATTTAGGAA AΝTTAAGTCC CGAAGCACAA CTTGCGGCTG 851 CAACCGCATT ACAAGACAGT GCTTTTGCGG TAAAAGACGG TATCAATTCC 901 GCCAGACAAT GGGCTGATGC CCATCCGAAT ATAACTGCAA CAGCCCAAAC 951 TGCCCTTGCC GTAGCAGANG CCGCAACTAC GGTTTGGGGC GGTAAAAAAG 1001 TAGAACTTAA CCCGACCAAA TGGGATTGGG TTAAAAATAC NGGCTATAAN 1051 ACACCTGCTG TTCGCACCAT GCATACTTTG GATGGGGAAA TGGCCGGTGG 1101 GAATAGACCG CCTAAATCTA TAACGTCCAA CAGCAAAGCA GATGCTTCCA 1151 CACAACCGTC TTTACAAGCG CAACTAATTG GAGAACAAAT TANNNNNGGG 1201 CATGCTTATA ACAAGCATGT CATAAGACAA CAAGAATTTA CGGATTTAAA 1251 TATCAATTCA CCAGCAGATT TTGCTCGGCA TATTGAAAAT ATTGTTAGCC 1301 ATCCANCAAA TATGAAAGAG TTACCTCGCG GTAGAACTGC GTATTGGGAT 1351 NATAAAACAG GGACNATAGT TATCCGAGAT AAAAATTCTG ACGATGGAGG 11440011 TTAACCAAGGCCAATTTTTT AAGGAACCCCAAAACCAATT CAGGTAAAAA ATATTATGAT GATTTATAG This encodes a protein having amino acid sequence <SEQ ID 168>: 1 MNXPIQKFMM LFAAAISXLQ IPISHANGLD ARLRDDMQAK HYEPGGKYHL 51 FGNARGSVKN RVYAVQTFDA TAVGPILPIT HERTGFEGII GYETHFSGHG 101 HEVHSPFDNH DSKSTSDFSG GVDGGFTVYQ LHRTGSEIHP EDGYDGPQGS 151 DYPPPGGARD IYXXYVKGTS TKTKSNIVPR APFSDRWLKE NAGAASGFFS 201 RADEAGKLIW ESDPNKNWWA NRMDDIRGIV QGAVNPFLMG FQGVGIGAIT 251 DSAVSPVTDT AAQQTLQGXN HLGXLSPEAQ LAAATALQDS AFAVKDGINS 301 ARQWADAHPN ITATAQTALA VAXAATTVWG GKKVELNPTK WDWVKNTGYX 351 TPAVRTMHTL DGEMAGGNRP PKSITSNSKA DASTQPSLQA QLIGEQIXXG 401 HAYNKHVIRQ QEFTDLNINS PADFARHIEN IVSHPXNMKE LPRGRTAYWD 451 XKTGTIVIRD KNSDDGGTAF RPTSGKKYYD DL* ORF29a and ORF29-1 show 90.1% identity in 385 aa overlap: 10 20 30 40 50 60 orf29a. pep MNXPIQKFMMLFAAAISXLQIPISHANGLDARLRDDMQAKHYEPGGKYHLFGNARGSVKN II II I II I II II I II I I II I II I II II I I I II II I II I I II II II II II I II I I I I I : orf29-1 MNLPIQKFMMLFAAAISLLQIPISHANGLDARLRDDMQAKHYEPGGKYHLFGNARGSVKK 10 20 30 40 50 60 70 80 90 100 110 120 orf29a. pep RVYAVQTFDATAVGPILPITHERTGFEGIIGYETHFSGHGHEVHSPFDNHDSKSTSDFSG II I II II I II II I : I : II I II I II I I II : I I I II I I I II I I II II II I : I I II II I I I I I orf29-1 RVYAVQTFDATAVSPVLPITHERTGFEGVIGYETHFSGHGHEVHSPFDHHDSKSTSDFSG 70 80 90 100 110 120 130 140 150 160 170 180 orf29a. pep GVDGGFTVYQLHRTGSEIHPEDGYDGPQGSDYPPPGGARDIYXXYVKGTSTKTKSNIVPR I II I I I II I II I II I I I II I I II I II II II II I II I I I II I I II II II II I I: I I I I : orf29-1 GVDGGFTVYQLHRTGSEIHPEDGYDGPQGSDYPPPGGARDIYSYYVKGTSTKTKTNIVPQ 130 140 150 160 170 180 190 200 210 220 230 240 orf29a . pep APFSDRWLKENAGAASGFFSRADEAGKLIWESDPNKNWWANRMDDIRGIVQGAVNPFLMG I I II II I I II I I I II II I II II II II II II I II II II II I I II IM II II I I II II I I I I orf29-1 APFSDRWLKENAGAASGFFSRADEAGKLIWESDPNKNWWANRMDDVRGIVQGAVNPFLMG 190 200 210 220 230 240 250 260 270 280 290 300 orf29a. pep FQGVGIGAITDSAVSPVTDTAAQQTLQGXNHLGXLSPEAQLAAATALQDSAFAVKDGINS II II I II I II I II II I I II I I II II II I I II II I II II I I I: II I I II I I I I I I I I orf29-1 FQGVGIGAITDSAVSPVTDTAAQQTLQGINDLGKLSPEAQLAAASLLQDSAFAVKDGINS 250 260 270 280 290 300 310 320 330 340 350 360 orf29a . pep ARQWADAHPNITATAQTALAVAXAATTVWGGKKVELNPTKWDWVKNTGYXTPAVRTMHTL I : M II I I II I II I II I II : : I I I I I I I II II I I I II II II I I II I II : I I : I I orf29-1 AKQWADAHPNITATAQTALSAAEAAGTVWRGKKVELNPTKWDWVKNTGYKKPAARHMQTL 310 320 330 340 350 360 370 380 390 400 410 420 orf29a. pep DGEMAGGNRPPKSITSNSKADASTQPSLQAQLIGEQIXXGHAYNKHVIRQQEFTDLNINS I I I I I I IM I I I: M I orf29-1 DGEMAGGNKPIKSLP-NSAAEKRKQNFEKFNSNWSSASFDSVHKTLTPNAPGILSPDKVK 370 380 390 400 410 Homology with a predicted ORF from N.gonorrhoeae ORF29 shows 88.8% identity over a 125aa overlap with a predicted ORF (ORF29.ng) from N gonorrhoeae: orf29. pep VSPVLPITHERTGFEGVIGYETHFSGHGHE 30 I : I: I II II II II II II I II I I II I I I II I orf29ng EPGGKYHLFGΝARGSVKΝRVCAVQTFDATAVGPILPITHERTGFEGVIGYETHFSGHGHE 102 orf29. pep VHSPFDHHDSKSTSDFSGGVDGGFTVYQLHRTWSEIHPEDEYDGPQAAXYPPPGGARDIY 90 I II I II : I II II II II I II II I II II I II I I I I I II II I I I II M: I I II I I I I II I orf29ng VHSPFDΝHDSKSTSDFSGGVDGGFTVYQLHRTGSEIHPEDGYDGPQGGGYPPPGGARDIY 162 orf29. pep SYYVKGTSTKTKTSIVPQAPFSDRWLEEΝAGAASG 125 I M : I I I I I I I I : II I I II I II I I : II II II I I orf29ng SYHIKGTSTKTKIΝTVPQAPFSDRWLKEΝAGAASGFLSRADEAGKLIWEΝDPDKΝWRAΝR 222 The complete length ORF29ng nucleotide sequence <SEQ LD 169> is predicted to encode a protein having amino acid sequence <SEQ ID 170>: 1 MΝLPIQKFMM LFAAAISLLQ IPISHAΝGLD ARLRDDMQAK HYEPGGKYHL 51 FGΝARGSVKΝ RVCAVQTFDA TAVGPILPIT HERTGFEGVI GYETHFSGHG 101 HEVHSPFDΝH DSKSTSDFSG GVDGGFTVYQ LHRTGSEIHP EDGYDGPQGG 151 GYPPPGGARD IYSYHIKGTS TKTKIΝTVPQ APFSDRWLKE ΝAGAASGFLS 201 RADEAGKLIW EΝDPDKΝWRA ΝRMDDIRGIV QGAVΝPFLTG FQGLGVGAIT 251 DSAVSPVTYA AARKTLQGIH ΝLGΝLSPEAQ LAAATALQDS AFAVKDSIΝS 301 ARQWADAHPΝ ITATAQTALA VTEAATTVWG GKKVELΝPAK WDWVKΝTGYK 351 KPAARHMQTV DGEMAGGΝKP LESKΝTVTTΝ ΝFFEΝTGYTE KVLRQASΝGD 401 YHGFPQSVDA FSEΝGTVIQI VGGDΝIVRHK LYIPGSYKGK DGΝFEYIREA 451 DGKIΝHRLFV PΝQQLPEK* In a second experiment, the following DΝA sequence <SEQ ID 171> was identified: 1 atgAATTTGC CTATTCAAAA ATTCATGATG ctgttggcAg cggcaatatc 51 gatgctGCat ATCCCCATTA GTCATGCGAA CGGTTTGGAT GCCCGTTTGC 101 GCGATGATAT GCAGGCAAAA CACTACGAAC CGGGTGGCAA ATACCATCTG 151 TTTGGTAATG CTCGCGGCAG TGTTAAAAAT CGGGTTTGCG CCGTCCAAAC 201 ATTTGATGCA ACTGCGGTCG GCCCCATACT GCCTATTACA CACGAACGGA 251 CAGGATTTGA AGGTGTTATC GGCTATGAAA CCCATTTTTC AGGACACGGA 301 CACGAAGTAC ACAGTCCGTT CGATAATCAT GATTCAAAAA GCACTTCTGA 351 TTTCAGCGGC GGCGTAGACG GCGGTTTTAC CGTTTACCAA CTTCATCGGA 401 CAGGGTCGGA AATACATCCC GCAGACGGAT ATGACGGGCC TCAAGGCGGC 451 GGTTATCCGG AACCACAAGG GGCAAGGGAT ATATACAGCT ACCATATCAA 501 AGGAACTTCA ACCAAAACAA AGATAAACAC TGTTCCGCAA GCCCCTTTTT 551 CAGACCGCTG GCTAAAAGAA AATGCCGGTG CCGCTTCCGG TTTTCTCAGC 601 CGTGCGGATG AAGCAGGAAA ACTGATATGG GAAAACGACC CCGATAAAAA 651 TTGGCGGGCT AACCGTATGG ATGATATTCG CGGCATCGTC CAAGGTGCGG 701 TTAATCCTTT TTTAACGGGT TTTCAAGGGG TAGGGATTGG GGCAATTACA 751 GACAGTGCGG TAAGCCCGGT CACAGATACA GCCGCTCAGC AGACTCTACA 801 AGGTATTAAT GATTTAGGAA ATTTAAGTCC GGAAGCACAA CTTGCCGCCG 851 CGAGCCTATT ACAGGACAGT GCCTTTGCGG TAAAAGACGG CATCAATTCC 901 GCCAGACAAT GGGCTGATGC CCATCCGAAT ATAACAGCAA CAGCCCAAAC 951 TGCCCTTGCC GTAGCAGAGG CCGCAGGTAC GGTTTGGCGC GGTAAAAAAG 1001 TAGAACTTAA CCCGACCAAA TGGGATTGGG TTAAAAATAC CGGCTATAAA 1051 AAACCTGCTG CCCGCCATAT GCAGACTGTA GATGGGGAGA TGGCAGGGGG 1101 GAATAGACCG CCTAAATCTA TAACGTCGGA AGGAAAAGCT AATGCTGCAA 1151 CCTATCCTAA GTTGGTTAAT CAGCTAAATG AGCAAAACTT AAATAACATT 1201 GCGGCTCAAG ATCCAAGATT GAGTCTAGCT ATTCATGAGG GTAAAAAAAA 1251 TTTTCCAATA GGAACTGCAA CTTATGAAGA GGCAGATAGA CTAGGTAAAA 1301 TTTGGGTTGG TGAGGGTGCA AGACAAACTA GTGGAGGCGG ATGGTTAAGT 1351 AGAGATGGCA CTCGACAATA TCGGCCACCA ACAGAAAAAA AATCACAATT 1401 TGCAACTACA GGTATTCAAG CAAATTTTGA AACTTATACT ATTGATTCAA 1451 ATGAAAAAAG AAATAAAATT AAAAATGGAC ATTTAAATAT TAGGTAA This encodes a protein having amino acid sequence <SEQ ID 172; ORF29ng-l>: 1 MΝLPIQKFMM LLAAAISMLH IPISHAΝGLD ARLRDDMQAK HYEPGGKYHL 51 FGΝARGSVKΝ RVCAVQTFDA TAVGPILPIT HERTGFEGVI GYETHFSGHG 101 HEVHSPFDNH DSKSTSDFSG GVDGGFTVYQ LHRTGSEIHP ADGYDGPQGG 151 GYPEPQGARD IYSYHIKGTS TKTKINTVPQ APFSDRWLKE NAGAASGFLS 201 RADEAGKLIW ENDPDKNWRA NRMDDIRGIV QGAVNPFLTG FQGVGIGAIT 251 DSAVSPVTDT AAQQTLQGIN DLGNLSPEAQ LAAASLLQDS AFAVKDGINS 301 ARQWADAHPN ITATAQTALA VAEAAGTVWR GKKVELNPTK WDWVKNTGYK 351 KPAARHMQTV DGEMAGGNRP PKSITSEGKA NAATYPKLVN QLNEQNLNNI 401 AAQDPRLSLA IHEGKKNFPI GTATYEEADR LGKIWVGEGA RQTSGGGWLS 451 RDGTRQYRPP TEKKSQFATT GIQANFETYT IDSNEKRNKI KNGHLNIR* ORF29ng-l and ORF29-1 show 86.0% identity in 401 aa overlap: 10 20 30 40 50 60 orf29ng-l.pep MNLPIQKFMMLLAAAISMLHIPISHANGLDARLRDDMQAKHYEPGGKYHLFGNARGSVKN I I I II I II II M II I I I : I : II II I I II II I II II II II I I II II I II I II I II I II I I : orf29-1 MNLPIQKFMMLFAAAISLLQIPISHANGLDARLRDDMQAKHYEPGGKYHLFGNARGSVKK 10 20 30 40 50 60 70 80 90 100 110 120 orf29ng-l.pep RVCAVQTFDATAVGPILPITHERTGFEGVIGYETHFSGHGHEVHSPFDNHDSKSTSDFSG II II II II I II I : I: I I II II I I I I I I II I II II II I I II II II I I I: I II I I I I II I I orf29-1 RVYAVQTFDATAVSPVLPITHERTGFEGVIGYETHFSGHGHEVHSPFDHHDSKSTSDFSG 70 80 90 100 110 120 130 140 150 160 170 180 orf29ng-l.pep GVDGGFTVYQLHRTGSEIHPADGYDGPQGGGYPEPQGARDIYSYHIKGTSTKTKINTVPQ I II II I II I II II II II II I I II I II I I : II I I II I II II :: I II I II I I I III orf29-1 GVDGGFTVYQLHRTGSEIHPEDGYDGPQGSDYPPPGGARDIYSYYVKGTSTKTKTNIVPQ 130 140 150 160 170 180 190 200 210 220 230 240 orf29ng-l.pep APFSDRWLKENAGAASGFLSRADEAGKLIWENDPDKNWRANRMDDIRGIVQGAVNPFLTG II II I II II I II I I II I I : I I II I I I II II I : I I : II I II II II : I I II II I I I I II I orf29-1 APFSDRWLKENAGAASGFFSRADEAGKLIWESDPNKNWWANRMDDVRGIVQGAVNPFLMG 190 200 210 220 230 240 250 260 270 280 290 300 orf29ng-l .pep FQGVGIGAITDSAVSPVTDTAAQQTLQGINDLGNLSPEAQLAAASLLQDSAFAVKDGINS II II I II II I II I I I I II I I I II I II II I I II I : I I I I II I I I I I I I I II II I I I I I II I orf29-1 FQGVGIGAITDSAVSPVTDTAAQQTLQGINDLGKLSPEAQLAAASLLQDSAFAVKDGINS 250 260 270 280 290 300 310 320 330 340 350 360 orf29ng-l . pep ARQWADAHPNITATAQTALAVAEAAGTVWRGKKVELNPTKWDWVKNTGYKKPAARHMQTV I : I I I I I I I I M I I I I I I MM II I I I I I I I I II I I I I I I II II I I I I II I II II I I I I : orf29-1 AKQWADAHPNITATAQTALSAAEAAGTVWRGKKVELNPTKWDWVKNTGYKKPAARHMQTL 310 320 330 340 350 360 370 380 390 400 410 419 orf29ng-l.pep DGEMAGGNRPPKSI-TSEGKANAATYPKLVNQLNEQNLNNIAAQDPRLSLAIHEGKKNFP II II I I I I: I II: : I :: :: | : :: : ::::: orf29-1 DGEMAGGNKPIKSLPNSAAEKRKQNFEKFNSNWSSASFDSVHKTLTPNAPGILSPDKVKT 370 380 390 400 410 420 420 430 440 450 460 470 479 orf29ng-l.pep IGTATYEEADRLGKIWVGEGARQTSGGGWLSRDGTRQYRPPTEKKSQFATTGIQANFETY orf29-1 RYTSLDGKITIIKDNENNYFRIHDNSRKQYLDSNGNAVKTGNLQGKQAKDYLQQQTHIRN 430 440 450 460 470 480 Based on this analysis, including the presence of a putative leader sequence in the gonococcal protein, it is predicted that the proteins from N. meningitidis .and N.gonorrhoeae, .and their epitopes, could be useful antigens for vaccines or diagnostics, or for raising antibodies. Example 21 The following partial DNA sequence was identified in N. meningitidis <SEQ ID 173>: 1 ATGAAAAAAC AAATCACCGC AGCCGTAATG ATGCTGTCTA TGATTGCCCC 51 CGCAATGGCA AACGGCTTGG ACAATCAGGC ATTTGAAGAC CAAATGTTCC 101 ACACGCGGGC AGATGCACCG ATGCAG ... This corresponds to the amino acid sequence <SEQ ID 174; ORF30>: 1 MKKQITAAVM MLSMIAPAMA NGLDNQAFED QMFHTRADAP MQ.. Further work revealed the complete nucleotide sequence <SEQ ID 175>: 1 ATGAAAAAAC AAATCACCGC AGCCGTAATG ATGCTGTCTA TGATTGCCCC 51 CGCAATGGCA AACGGCTTGG ACAATCAGGC ATTTGAAGAC CAAGTGTTCC 101 ACACGCGGGC AGATGCACCG ATGCAGTTGG CGGAGCTTTC TCAAAAGGAG 151 ATGAAGGAGA CAGAGGGGGC GTTTCTTCCA TTGGCTATCT TGGGTGGTGC 201 TGCCATTGGT ATGTGGACAC AGCATGGTTT TAGTTATGCA ACGACAGGCA 251 GACCAGCTTC TGTTAGAGAT GTTGCTATTG CTGGCGGATT AGGCGCAATT 301 CCTGGTGGTG TAGGCGCCGC AGGAAAGGTT GTTTCCTTTG CTAAATATGG 351 ACGTGAGATT AAAATCGGCA ATAATATGCG GATAGCCCCT TTCGGTAATA 401 GAACAGGTCA TCCTATTGGA AAATTTCCCC ATTATCATCG TCGAGTTACG 451 GATAATACGG GCAAGACTTT GCCTGGACAG GGAATTGGTC GTCATCGCCC 501 TTGGGAATCA AAATCTACGG ACAGATCATG GAAAAACCGC TTCTAA This corresponds to the amino acid sequence <SEQ ID 176; ORF30-1>: 1 MKKQITAAVM MLSMIAPAMA NGLDNQAFED QVFHTRADAP MQiAELSQKE 51 MKETEGAFLP LAILGGAAIG MWTQHGFSYA TTGRPASVRD VAIAGGLGAI 101 PGGVGAAGKV VSFAKYGREI KIGNNMRIAP FGNRTGHPIG KFPHYHRRVT 151 DNTGKTLPGQ GIGRHRPWES KSTDRSWKNR F* Computer analysis of this amino acid sequence gave the following results: Homology with a predicted ORF from N.meninsitidis (strain Al ORF30 shows 97.6% identity over a 42aa overlap with an ORF (ORF30a) from strain A of N meningitidis: 10 20 30 40 orf30. pep MKKQITAAVMMLSMIAPAMAΝGLDΝQAFEDQMFHTRADAPMQ I I II II II I II II I II II I II I II II I II II : II I II II II I orf30a MKKQITAAVMMLSMIAPAMAΝGLDΝQAFEDQVFHTRADAPMQLAELSQKEMKXTXGAFLP 10 20 30 40 50 60 orf30a LXILGGAAIGMWTQHGFSYATTGRPASVRDVAIAGGLGAIPGXVGAAGKWSFAKYGREI 70 80 90 100 110 120 The complete length ORF30a nucleotide sequence <SEQ ID 177> is: 1 ATGAAAAAAC AAATCACCGC AGCCGTAATG ATGCTGTCTA TGATTGCCCC 51 CGCAATGGCA AACGGCTTGG ACAATCAGGC ATTTGAAGAC CAAGTGTTCC 101 ACACGCGGGC AGATGCACCG ATGCAGTTGG CGGAGCTTTC TCAAAAGGAG 151 ATGAAGGAΝA CAGΝGGGGGC GTTTCTTCCA TTGGΝTATCT TGGGTGGTGC 201 TGCCATTGGT ATGTGGACAC AGCATGGTTT TAGTTATGCA ACGACAGGCA 251 GACCAGCTTC TGTTAGAGAT GTTGCTATTG CTGGCGGATT AGGCGCAATT 301 CCTGGTGΝTG TAGGCGCCGC AGGAAAGGTT GTTTCCTTTG CTAAATATGG 351 ACGTGAGATT AAAATCGGCA ATAATATGCG GATAGCCCCT TTCGGTAATA 401 GAACAGGTCA TCCTATTGGΝ AAATTTCCCC ATTATCATCG TCGAGTTACG 451 GATAATACGG GCAAGACTTT GCCTGGACAG GGAATTGGTC GTCATCGCCC 501 TTGGGAATCA AAATCTACGG ACAGATCATG GAAAAACCGC TTCTAA This encodes a protein having amino acid sequence <SEQ LD 178>: 1 MKKQITAAVM MLSMIAPAMA ' NGLDNQAFED QVFHTRADAP MQLAELSQKE 51 MKXTXGAFLP LXILGGAAIG MWTQHGFSYA TTGRPASVRD VAIAGGLGAI 101 PGXVGAAGKV VSFAKYGREI KIGNNMRIAP FGNRTGHPIG KFPHYHRRVT 151 DNTGKTLPGQ GIGRHRPWES KSTDRSWKNR F* ORF30a and ORF30-1 show 97.8% identity in 181 aa overlap: orf 30a . pep MKKQITAAVMMLSMIAPAMANGLDNQAFEDQVFHTRADAPMQLAELSQKEMKXTXGAFLP 60 I II II I II II I II II I II II I II II I II I II I II I I I II I I II II II I I I II I I I I II orf30-1 MKKQITAAVMMLSMIAPAMANGLDNQAFEDQVFHTRADAPMQLAELSQKEMKETEGAFLP 60 orf30a .pep LXILGGAAIGMWTQHGFSYATTGRPASVRDVAIAGGLGAIPGXVGAAGKWSFAKYGREI 120 I I II I II II I II II I I I I I I II I II II II II I I II II II I I I I II I II I II I II II I I orf30-1 LAILGGAAIGMWTQHGFSYATTGRPASVRDVAIAGGLGAIPGGVGAAGKWSFAKYGREI 120 orf30a. pep KIGNNMRIAPFGNRTGHPIGKFPHYHRRVTDNTGKTLPGQGIGRHRPWESKSTDRSWKNR 180 I I I I II II II II II I II II II I II I II I I II II I I II II I II II II I II I II I I I I I I II orf30-l KIGNNMRIAPFGNRTGHPIGKFPHYHRRVTDNTGKTLPGQGIGRHRPWESKSTDRSWKNR 180 orf30a.pep FX I I orf30-l FX Homology with a predicted ORF from N.sonorrhoeae ORF30 shows 97.6% identity over a 42aa overlap with a predicted ORF (ORF30.ng) from N gonorrhoeae: orf30. pep MKKQITAAVMMLSMIAPAMAΝGLDΝQAFEDQMFHTRADAPMQ 42 I I II II I I II II II I I II II II II I I II I I I : I II I I II II I orf30ng MKKQITAAVMMLSMIAPAMAΝGLDΝQAFEDQVFHTRADAPMQLAELSQKEMKETEGAFLP 60 The complete length ORF30ng nucleotide sequence <SEQ ID 179> is 1 ATGAAAAAAC AAATCACCGC AGCCGTAATG ATGCTGTCTA TGATCGCCCC 51 CGCAATGGCA AACGGATTGG ACAATCAGGC ATTTGAAGAC CAAGTGTTCC 101 ACACGCGGGC AGATGCGCCG ATGCAGTTGG CGGAGCTTTC TCAGAAGGAG 151 ATGAAGGAGA CTGAAGGGGC TTTTCTTCCA TTGGCTATCT TGGGTGGTGC 201 TGCCATTGGT ATGTGGACAC AGCATGGTTT TAGTTATGCA ACGACAGGCA 251 GACCAGCTTC TGTTAGAGAT GTTGCTGGCG GATTAGGCGC AATTCCTGGT 301 GATGTAGGTG CTGCAGGAAA GGTTGTTTCC TTTGCTAAAT ATGGACGTGA 351 GATTAAAATC GGCAATAATA TGCGGATAGC CCCTTTCGGT AATAGAACAG 401 GTCATCCTAT TGGAAAATTT CCCCATTATC ATCGTCGAGT TACGGATAAT 451 ACGGGCAAGA CTTTGCCTGG ACAGGGAATT GGTCGTCATC GCCCTTGGGA 501 ATCAAAATCT ACGGACAGAT CATGGAAAAA CCGCTTCTAA This encodes a protein having amino acid sequence <SEQ ID 180>: 1 MKKQITAAVM MLSMIAPAMA NGLDNQAFED QVFHTRADAP MQLAELSQKE 51 MKETEGAFLP LAILGGAAIG MWTQHGFSYA TTGRPASVRD VAGGLGAIPG 101 DVGAAGKWS FAKYGREIKI GNNMRIAPFG NRTGHPIGKF PHYHRRVTDN 151 TGKTLPGQGI GRHRPWESKS TDRSWKNRF* ORF30ng and ORF30-1 show 98.3% identity in 181 aa overlap: 10 20 30 40 50 60 orf30ng . pep MKKQITAAVMMLSMIAPAMANGLDNQAFEDQVFHTRADAPMQLAELSQKEMKETEGAFLP II I II I I I I I I II II I II II I II I I II I I I I I II I II I I I I II II II II I II I II I I I I I orf30-1 MKKQITAAVMMLSMIAPAMANGLDNQAFEDQVFHTRADAPMQLAELSQKEMKETEGAFLP 10 20 30 40 50 60 70 80 90 100 110 orf30ng.pep LAILGGAAIGMWTQHGFSYATTGRPASVRDVA—GGLGAIPGDVGAAGKWSFAKYGREI I I I I I I I I II II I I I I I II I I I I I II II I II I I II I II II I II I I I II II I II I II I orf30-1 LAILGGAAIGMWTQHGFSYATTGRPASVRDVAIAGGLGAIPGGVGAAGKWSFAKYGREI 70 80 90 100 110 120 120 130 140 150 160 170 orf30ng.pep KIGNNMRIAPFGNRTGHPIGKFPHYHRRVTDNTGKTLPGQGIGRHRPWESKSTDRSWKNR II I II I II I II II II I I I I II I II I I II I I I I II I I II I I I I I II I I II I II I I I I II I I orf30-1 KIGNNMRIAPFGNRTGHPIGKFPHYHRRVTDNTGKTLPGQGIGRHRPWESKSTDRSWKNR 130 140 150 160 170 180 180 orf30ng.pep FX II orf30-l FX Based on this analysis, including the presence of a putative leader sequence in the gonococcal protein, it is predicted that the proteins from N. meningitidis .and N.gonorrhoeae, .and their epitopes, could be useful .antigens for vaccines or diagnostics, or for raising antibodies. Example 22 The following partial DNA sequence was identified in N. meningitidis <SEQ ID 181>: 1 ATGAATAAAA CTCTCTATCG TGTAATTTTC AACCGCAAAC GTGGGGCTGT 51 GrTAGCCGTT GCTGAAACTA CCAAGCGCGA AGGTAAAAGC TGTGCCGATA 101 GTGATTCAGG CAGCGCTCAT GTGAAATCTG TTCCTTTTGG TACTACTCAT 151 GCACCTGTTT GTg.CGTTaC AAATATCTTT TCTTTTTCTT TATTGGGCTT 201 TTCTTTATGT TTGGCTGTAG GtacGGyCAA TATTGCTTTT GCTGATGGCA 251 TT.. This corresponds to the amino acid sequence <SEQ ID 182; ORF31>: 1 MNKTLYRVIF NRKRGAVXAV AETTKREGKS CADSDSGSAH VKSVPFGTTH 51 APVCXVTNIF SFSLLGFSLC LAVGTXNIAF ADGI .. Further work revealed a further partial nucleotide sequence <SEQ ID 183>: 1 ATGAATAAAA CTCTCTATCG TGTAATTTTC AACCGCAAAC GTGGGGCTGT 51 GGTAGCCGTT GCTGAAACTA CCAAGCGCGA AGGTAAAAGC TGTGCCGATA 101 GTGATTCAGG CAGCGCTCAT GTGAAATCTG TTCCTTTTGG TACTACTCAT 151 GCACCTGTTT GTCGTTCAAA TATCTTTTCT TTTTCTTTAT TGGGCTTTTC 201 TTTATGTTTG GCTGTAGGTA CGGCCAATAT TGCTTTTGCT GATGGCATT.. This corresponds to the amino acid sequence <SEQ ID 184; ORF31-l>: 1 MNKTLYRVIF NRKRGAWAV AETTKREGKS CADSDSGSAH VKSVPFGTTH 51 APVCRSNIFS FSLLGFSLCL AVGTANIAFA DGI .. Computer analysis of this amino acid sequence gave the following results: Homology with a predicted ORF from N.gonorrhoeae ORF31 shows 76.2% identity over a 84aa overlap with a predicted ORF (ORF31.ng) from N. gonorrhoeae: orf31. pep MNKTLYRVIFNRKRGAVXAVAETTKREGKSCADSDSGSAHVKSVPFGTTHAPVCXVTNIF 60 II I I II I I I I I II I I II I I II I I II I II II II I II I :: II I I I II :: I orf31ng MNKTLYRVIFNRKRGAWAVAETTKREGKSCADSGSGSVYVKSVSFIPTH SKAF 54 orf31. pep SFSLLGFSLCLAVGTXNIAFADGI 84 II I I II II I l I I II II I I I orf31ng CFSALGFSLCLALGTVNIAFADGIITDKAAPKTQQATILQTGNGIPQVNIQTPTSAGVSV 114 The complete length ORF3 lng nucleotide sequence <SEQ YD 185> is: 1 ATGAACAAAA CCCTCTATCG TGTGATTTTC AACCGCAAAC GCGGTGCTGT 51 GGTAGCTGTT GCCGAAACCA CCAAGCGCGA AGGTAAAAGC TGTGCCGATA 101 GTGGTTCGGG CAGCGTTTAT GTGAAATCCG TTTCTTTCAT TCCTACTCAT 151 TCCAAAGCCT TTTGTTTTTC TGCATTAGGC TTTTCTTTAT GTTTGGCTTT 201 GGGTACGGTC AATATTGCTT TTGCTGACGG CATTATTACT GATAAAGCTG 251 CTCCTAAAAC CCAACAAGCC ACGATTCTGC AAACAGGTaa cGGCATACCG 301 CAAGTCAATA TTCAAACCCC TACTTCGGCA GGGGTTTCTG TTAATCAATA 351 TGCCCAGTTT GATGTGGGTA ATCGCGGGGC GATTTTAAAC AACAGTCGCA 401 GCAACACCCA AACACAGCTA GGCGGTTGGA TTCAAGGCAA TCCTTGGTTG 451 ACAAGGGGCG AAGCACGTGT GGTTGTAAAC CAAATCAACA GCAGCCATCC 501 TTCACAACTG AATGGCTATA TTGAAGTGGG TGGACGACGT GCAGAAGTCG 551 TTATTGCCAA TCCGGCAGGG ATTGCAGTCA ATGGTGGTGG TTTTATCAAT 601 GCTTCCCGTG CCACTTTGAC GACAGGCCAA CCGCAATATC AAGCAGGAGA 651 CTTTAGCGGC TTTAAGATAA GGCAAGGCAA TGCTGTAATC GCCGGACACG 701 GTTTGGATGC CCGTGATACC GATTTCACAC GTATTCTTGT ATGCCAACAA 751 AATCACCTTG ATCAGTACGG CCGAACAAGC AGGCATTCGT AA This encodes a protein having amino acid sequence <SEQ ID 186>: 1 MNKTLYRVIF NRKRGAWAV AETTKREGKS CADSGSGSVY VKSVSFIPTH 51 SKAFCFSALG FSLCLALGTV NIAFADGIIT DKAAPKTQQA TILQTGNGIP 101 QVNIQTPTSA GVSVNQYAQF DVGNRGAILN NSRSNTQTQL GGWIQGNPWL 151 TRGEARVWN QINSSHPSQL NGYIEVGGRR AEWIANPAG IAVNGGGFIN 201 ASRATLTTGQ PQYQAGDFSG FKIRQGNAVI AGHGLDARDT DFTRILVCQQ 251 NHLDQYGRTS RHS* This gonococcal protein shares 50% identity over a 149aa overlap with the pore-forming hemolysins-like HecA protein from Erwinia chrysanthemi (accession number L39897): orf31ng 96 GNGIPQVNIQTPTSAGVSVNQYAQFDVGNRGAILNNSRSN-TQTQLGGWIQGNPWLTRGE 154 GNG+P VNI TP ++G+S N+Y F+V NRG ILNN + T +QLGG IQ NP L HecA 45 GNGVPWNIATPDASGLSHNRYHDFNVDNRGLILNNGTARLTPSQLGGLIQNNPNLNGRA 104 Orf31ng 155 ARWVNQINSSHPSQLNGYIEVGGRRAEWIANPAGIAVNGGGFINASRATLTTGQPQYQ 214 A ++N++ S + S+L GY+EV G+ A VV+ANP GI +G GF+N R TLTTG PQ+ HecA 105 AAAILNEWSPNRSRLAGYLEVAGQAANVWANPYGITCSGCGFLNTPRLTLTTGTPQFD 164 Orf31ng 215 -AGDFSGFKIRQGNAVIAGHGLDARDTDF 242 AG SG +R G+ +1 G GLDA +D+ HecA 165 AAGGLSGLDVRGGDILIDGAGLDASRSDY 193 Furthermore, ORF31ng .and ORF31-1 show 79.5% identity in 83 aa overlap: 10 20 30 40 50 60 orf31-1.pep MNKTLYRVIFNRKRGAWAVAETTKREGKSCADSDSGSAHVKSVPFGTTHAPVCRSNIFS II I I II I I I II I I I I I II II II II II I II I I I II II M: II I I I II M I orf31ng MNKTLYRVIFNRKRGAWAVAETTKREGKSCADSGSGSVYVKSVSFIPTH SKAFC 10 20 30 40 50 70 80 orf31-1. pep FSLLGFSLCLAVGTANIAFADGI II I I II II IM I M II I I I I I I orf31ng FSALGFSLCLALGTVNIAFADGIITDKAAPKTQQATILQTGNGIPQVNIQTPTSAGVSVN 60 70 80 90 100 110 On this basis, including the homology with hemolysms, and also with adhesins, it is predicted that the proteins from N. meningitidis and N.gonorrhoeae, and their epitopes, could be useful antigens for vaccines or diagnostics, or for raising antibodies. Example 23 The following partial DNA sequence was identified in N. meningitidis <SEQ ID 187>: 1 ATGAATACTC CTCCTTTTGT CTGTTGGATT TTTTGCAAGG TCATCGACAA 51 TTTCGGCGAC ATCGGCGTTT CGTGGCGGCT CGCCCGTGTT TTGCACCGCG 101 AACTCGGTTG GCAGGTGCAT TTGTGGACGG ACGATGTGTC CGCCTTGCGT 151 GCGCTTTGCC CTGATTTGCC CGATGTTCCC TGCGTTCATC AGGATATTCA 201 TGTCCGCACT TGGCATTCCG ATGCGGCAGA TATTGATACC GCG.. This corresponds to the .amino acid sequence <SEQ ID 188; ORF32>: 1 MNTPPFVCWI FCKVIDNFGD IGVSWRLARV LHRELGWQVH LWTDDVSALR 51 ALCPDLPDVP CVHQDIHVRT WHSDAADIDT A.. Further work revealed the complete nucleotide sequence <SEQ ID 189>: 1 ATGAATACTC CTCCTTTTGT CTGTTGGATT TTTTGCAAGG TCATCGACAA 51 TTTCGGCGAC ATCGGCGTTT CGTGGCGGCT CGCCCGTGTT TTGCACCGCG 101 AACTCGGTTG GCAGGTGCAT TTGTGGACGG ACGATGTGTC CGCCTTGCGT 151 GCGCTTTGCC CTGATTTGCC CGATGTTCCC TGCGTTCATC AGGATATTCA 201 TGTCCGCACT TGGCATTCCG ATGCGGCAGA TATTGATACC GCGCCTGTTC 251 CCGATGTCGT CATCGAAACT TTTGCCTGCG ACCTGCCCGA AAATGTGCTG 301 CACATTATCC GCCGACACAA GCCGCTTTGG CTGAATTGGG AATATTTGAG 351 CGCGGAGGAA AGCAATGAAA GGCTGCATCT GATGCCTTCG CCGCAGGAGG 401 GTGTTCAAAA ATATTTTTGG TTTATGGGTT TCAGCGAAAA AAGCGGCGGG 451 TTGATACGCG AACGTGATTA CTGCGAAGCC GTCCGTTTCG ATACTGAAGC 501 CCTGCGAGAG CGGCTGATGC TGCCCGAAAA AAACGCCTCC GAATGGCTGC 551 TTTTCGGCTA TCGGAGCGAT GTTTGGGCAA AGTGGCTGGA AATGTGGCGA 660011 CAGGCAGGCA GCCCGATGAC ACTGTTGCTG GCGGGGACGC AAATCATCGA 651 CAGCCTCAAA CAAAGCGGCG TTATTCCGCA AGATGCCCTG CAAAACGACG 701 GCGATGTTTT TCAGACGGCA TCCGTCCGCC TCGTCAAAAT CCCTTTCGTG 751 CCGCAACAGG ACTTCGACCA ACTGCTGCAC CTTGCCGACT GCGCCGTCAT 801 CCGCGGCGAA GACAGTTTCG TGCGCGCCCA GCTTGCGGGC AAACCCTTCT 885511 TTTGGCACAT CTACCCGCAA GACGAGAATG TCCATCTCGA CAAACTCCAC 901 GCCTTTTGGG ATAAGGCACA CGGTTTCTAC ACGCCCGAAA CCGTGTCGGC 951 ACACCGCCGT CTTTCGGACG ACCTCAACGG CGGAGAGGCT TTATCCGCAA 1001 CACAACGCCT CGAATGTTGG CAAACCCTGC AACAACATCA AAACGGCTGG 1051 CGGCAAGGCG CGGAGGATTG GAGCCGTTAT CTTTTCGGGC AGCCGTCAGC 11110011 TCCTGAAAAA CTCGCTGCCT TTGTTTCAAA GCATCAAAAA ATACGCTAG This corresponds to the amino acid sequence <SEQ ID 190; ORF32-l>: 1 MNTPPFVCWI FCKVIDNFGD IGVSWRLARV LHRELGWQVH LWTDDVSALR 51 ALCPDLPDVP CVHQDIHVRT WHSDAADIDT APVPDWIET FACDLPENVL 101 HIIRRHKPLW LNWEYLSAEE SNERLHLMPS PQEGVQKYFW FMGFSEKSGG 151 LIRERDYCEA VRFDTEALRE RLMLPEKNAS EWLLFGYRSD VWAKWLEMWR 201 QAGSPMTLLL AGTQIIDSLK QSGVIPQDAL QNDGDVFQTA SVRLVKIPFV 251 PQQDFDQLLH LADCAVIRGE DSFVRAQLAG KPFFWHIYPQ DENVHLDKLH 301 AFWDKAHGFY TPETVSAHRR LSDDLNGGEA LSATQRLECW QTLQQHQNGW 351 RQGAEDWSRY LFGQPSAPEK LAAFVSKHQK IR* Computer analysis of this amino acid sequence gave the following results: Homology with a predicted ORF from N.meninsitidis (strain Al ORF32 shows 93.8% identity over a 81aa overlap with an ORF (ORF32a) from strain A of N. meningitidis: 10 20 30 40 50 60 orf32.pep MNTPPFVCWIFCKVIDNFGDIGVSWRLARVLHRELGWQVHLWTDDVSALRALCPDLPDVP I I I II I II I II I II II II I I II I II II II I I I II II I I II II II I I I II II II II orf32a MNTPPFSAGXFCKVIDNFGDIGVSWRLARVLHRELGWQVHLWTDDVSALRALCPDLPDVX 10 20 30 40 50 60 70 80 orf32. pep CVHQDIHVRTWHSDAADIDTA I I II I II II I II II I I I I II I orf32a CVHQDIHVRTWHSDAADIDTAPVXDWIETFACDLPENVLHIIRRHKPLWLXWEYLSAEX 70 80 90 100 110 120 The complete length ORF32a nucleotide sequence <SEQ ID 191> is: 1 ATGAATACTC CTCCTTTTTC TGCTGGANTT TTTTGCAAGG TCATCGACAA 51 TTTCGGCGAC ATCGGCGTTT CGTGGCGGCT TGCCCGTGTT TTGCACCGCG 101 AACTCGGTTG GCAGGTGCAT TTGTGGACGG ACGATGTGTC CGCCTTGCGT 151 GCGCTTTGCC CTGATTTGCC CGATGTTCNC TGCGTTCATC AGGATATTCA 201 TGTCCGCACT TGGCATTCCG ATGCGGCAGA TATTGATACC GCGCCTGTTC 251 NCGATGTCGT CATCGAAACT TTTGCCTGCG ACCTGCCCGA AAATGTGCTG 301 CACATCATCC GCCGACACAA GCCGCTTTGG CTGAANTGGG AATATTTGAG 351 CGCGGAGGAN AGCAATGAAA GGCTGCACNT GATGCCTTCG CCGCAGGAGA 401 GTGTTCNAAA ATANTTTTGG TTTATGGGTT TCAGCGAANN NAGCGGCGGA 451 CTGATACGCG AACGCGATTA CTGCGAAGCC GTCCGTTTCG ATAGCGGAGC 501 CTTGCGCAAG AGGCTGATGC TTCCCGAAAA AAACGNCCCC GAATGGCTGC 551 TTTTCGGCTA TCGGAGCGAT GTTTGGGCAA AGTGGCTGGA AATGTGGCGA 601 CAGGCAGGCA GTCCGTTGAC ACTTTTGCTG GCNGGGGCGC ANATTATCGA 651 CAGCCTCAAA CAAAACGGCG TTATTCCGCA AGATGCCCTG CAAAACGACG 701 GCGATGTTTT TCAGACGGCA TCCGTCCGCC TCGTCAAAAT CCCTTTCGTG 751 CCGCAACAGG ACTTCGACAA ACTGCTGCAC CTTGCCGACT GCGCCGTCAT 801 CCGCGGCGAA GACAGTTTCG TGCGCGCCCA GCTTGCGGGC AAACCCTTCT 851 TTTGGCACAT CTACCCGCAA GATGAGAATG TCCATCTCGA CAAACTCCAC 901 GCCTTTTGGG ATAAGGCACA CGGTTTCTAC ACGCCCGAAA CCGCATCGGC 951 ACACCGCCGC CTTTCAGACG ACCTCAACGG CGGAGAGGCT TTATCCGCAA 1001 CACAACGCCT CGAATGTTGG CAAATCCTGC AACAACATCA AAACGGCTGG 1051 CGGCAAGGCG CGGAGGATTG GAGCCGTTAT CTTTTTGGGC AGCCTTCCGC 1101 ATCCGAAAAA CTCGCCGCCT TTGTTTCAAA GCATCAAAAA ATACGCTAG This encodes a protein having amino acid sequence <SEQ ID 192>: 1 MNTPPFSAGX FCKVIDNFGD IGVSWRLARV LHRELGWQVH LWTDDVSALR 51 ALCPDLPDVX CVHQDIHVRT WHSDAADIDT APVXDWIET FACDLPENVL 101 HIIRRHKPLW LXWEYLSAEX SNERLHXMPS PQESVXKXFW FMGFSEXSGG 151 LIRERDYCEA VRFDSGALRK RLMLPEKNXP EWLLFGYRSD VWAKWLEMWR 201 QAGSPLTLLL AGAXIIDSLK QNGVIPQDAL QNDGDVFQTA SVRLVKIPFV 251 PQQDFDKLLH LADCAVIRGE DSFVRAQLAG KPFFWHIYPQ DENVHLDKLH 301 AFWDKAHGFY TPETASAHRR LSDDLNGGEA LSATQRLECW QILQQHQNGW 351 RQGAEDWSRY LFGQPSASEK LAAFVSKHQK IR* ORF32a and ORF32-1 show 93.2% identity in 382 aa overlap: 10 20 30 40 50 60 orf32-1. pep MNTPPFVCWIFCKVIDNFGDIGVSWRLARVLHRELGWQVHLWTDDVSALRALCPDLPDVP ' II II I I II II II I II I II II I II I II I I II I II II II I II II II I I I II II I I I I orf32a MNTPPFSAGXFCKVIDNFGDIGVSWRLARVLHRELGWQVHLWTDDVSALRALCPDLPDVX 10 20 30 40 50 60 70 80 90 100 110 120 orf32-1. pep CVHQDIHVRTWHSDAADIDTAPVPDWIETFACDLPENVLHIIRRHKPLWLNWEYLSAEE II I I II I I II I II II II I I II I I II I II II II I I I I I I I I I I I I I II I I I I II I I I I orf32a CVHQDIHVRTWHSDAADIDTAPVXDWIETFACDLPENVLHIIRRHKPLWLXWEYLSAEX 70 80 90 100 110 120 130 140 150 160 170 180 orf32-1. pep SNERLHLMPSPQEGVQKYFWFMGFSEKSGGLIRERDYCEAVRFDTEALRERLMLPEKNAS II I II I I II I I I : I I II I I I II I I I II II I I I I II I I I II : II I : I II I II I I orf32a SNERLHXMPSPQESVXKXFWFMGFSEXSGGLIRERDYCEAVRFDSGALRKRLMLPEKNXP 130 140 150 160 170 180 190 200 210 220 230 240 orf32-1. pep EWLLFGYRSDVWAKWLEMWRQAGSPMTLLLAGTQIIDSLKQSGVIPQDALQNDGDVFQTA I I II I I II I II I II II I II I I I II I: I II I I M II II I I I : I II II I I I II I II I I I II orf32a EWLLFGYRSDVWAKWLEMWRQAGSPLTLLLAGAXIIDSLKQNGVIPQDALQNDGDVFQTA 190 200 210 220 230 240 250 260 270 280 290 300 orf32-1. pep SVRLVKIPFVPQQDFDQLLHLADCAVIRGEDSFVRAQLAGKPFFWHIYPQDENVHLDKLH II I II I I I II I I I II I : I I II I I I II I I II I I II I II II II I II II I I I I I I II I II II I orf32a SVRLVKIPFVPQQDFDKLLHLADCAVIRGEDSFVRAQLAGKPFFWHIYPQDENVHLDKLH 250 260 270 280 290 300 310 320 330 340 350 360 orf32-1. pep AFWDKAHGFYTPETVSAHRRLSDDLNGGEALSATQRLECWQTLQQHQNGWRQGAEDWSRY M I II I I I II I II I : I I II I I I II I I II I I II I II II I II I I I I II I I I I II II I I I II orf 32a AFWDKAHGFYTPETASAHRRLSDDLNGGEALSATQRLECWQILQQHQNGWRQGAEDWSRY 310 320 330 340 350 360 370 380 orf 32-1 . pep LFGQPSAPEKLAAFVSKHQKIRX I I I I I I I I I I I I I I I I I I I I I I orf32a LFGQPSASEKLAAFVSKHQKIRX 370 380 Homology with a predicted ORF from N.gonorrhoeae ORF32 shows 95.1% identity over a 82aa overlap with a predicted ORF (ORF32.ng) from N. gonorrhoeae: orf 32 . pep MNTPPF-VCWIFCKVIDNFGDIGVSWRLARVLHRELGWQVHLWTDDVSALRALCPDLP 57 I I I I I I I I M I M I I M M I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I orf32ng MVMNTYAFPVCWIFCKVIDNFGDIGVSWRLARVLHRELGWQVHLWTDDVSALRALCPDLP 60 orf32.pep DVPCVHQDIHVRTWHSDAADIDTA 81 III II II I II II I II I I II I II I orf32ng DVPFVHQDIHVRTWHSDAADIDTAPVPDAVIETFACDLPENVLNIIRRHKPLWLNWEYLS 120 An ORF32ng nucleotide sequence <SEQ LD 193> was predicted to encode a protein having amino acid sequence <SEQ ID 194>: 1 MVMNTYAFPV CWIFCKVIDN FGDIGVSWRL ARVLHRELGW QVHLWTDDVS 51 ALRALCPDLP DVPFVHQDIH VRTWHSDAAD IDTAPVPDAV IETFACDLPE 101 NVLNIIRRHK PLWLNWEYLS AEESNERLHL MPSPQEGVQK YFWFMGFSEK 151 SGGLIRERDY REAVRFDTEA LRRRLVLPEK NAPEWLLFGY RGDVWAKWLD 201 MWQQAGSLMT LLLAGAQIID SLKQSGVIPQ NALQNEGGVF QTASVRLVKI 251 PFVPQQDFDK LLHLADCAVI RGEDSFVRTQ LAGKPFFWHI YPQDENVHLD 301 KLHAFWDKAY GFYTPETASV HRLLSDDLNG GEALSATQRL ECGVL* Further sequencing revealed the following DNA sequence <SEQ ID 195>: 1 ATGAATACAT ACGCTTTTCC TGTCTGTTGG ATTTTTTGCA AGGTCATCGA 51 CAATTTCGGC GACATCGGCG TTTCGTGGCG GCTCGCCCGT GTTTTGCACC 101 GCGAACTCGG TTGGCAGGTG CATTTGTGGA CGGACGACGT GTCCGCCTTG 151 CGCGCGCTTT GTCCCGATTT GCCCGATGTT CCCTTCGTTC ATCAGGATAT 201 TCATGTCCGC ACTTGGCATT CCGATGCGGC AGACATTGAT ACCGCGCCCG 251 TTCCCGATGC CGTTATCGAA ACTTTTGCCT GCGACCTGCC CGAAAATGTG 301 CTGAACATCA TCCGCCGACA CAAACCGCTT TGGCTGAATT GGGAATATTT 351 GAGCGCGGAG GAAAGCAATG AAAGGCTGCA CCTGATGCCT TCGCCGCAGG 401 AGGGCGTTCA AAAATATTTT TGGTTTATGG GTTTCAGCGA AAAAAGCGGC 451 GGGTTGATAC GCGAACGCGA TTACCGCGAA GCCGTCCGTT TCGATACCGA 501 AGCCCTGCGC CGGCGGCTGG TGCTGCCCGA AAAAAACGCC CCCGAATGGC 551 TGCTTTTCGG CTATCGGGGC GATGTTTGGG CAAAGTGGCT GGACATGTGG 601 CAACAGGCAG GCAGCCTGAT GACCCTACTG CTGGCGGGGG CGCAAATTAT 651 CGACAGCCTC AAACAAAGCG GCGTTATTCC GCAAAACGCC CTGCAAAAtg 701 aaggcgGTGT CTTTCagacG gcatccgTcC gccttGTCAA AAtcCCGTTC 751 GTGCcGCAAC AGGAcTTCGA CAAATTGCTG CAcctcgcCG ACTGCGCCGT 801 GATACGCGGC GAAGACAGTT TCGTGCGTAC CCAGCTTGCC GGAAAACCCT 851 TTTTTTGGCA CATCTACCCG CAAGACGAGA ATGTCCATCT CGACAAACTC 901 CACGCCTTTT GGGATAAGGC ATACGGCTTC TACACGCCCG AAACCGCATC 951 GGTGCACCGC CTCCTTTCGG ACGACCTCAA CGGCGGAGAG GCTTTATCCG 1001 CAACACAACG CCTCGAATGT TGGCAAACCC TGCAACAACA TCAAAACGGC 1051 TGGCGGCAAG GCGCGGAGGA TTGGAGCCGT TATCTTTTCG GGCAGCCTTC 1101 CGCATCCGAA AAACTCGCCG CCTTTGTTTC AAAGCATCAA AAAATACGCT 1151 AG This encodes a protein having amino acid sequence <SEQ ED 196; ORF32ng-l>: 1 MNTYAFPVCW IFCKVIDNFG DIGVSWRLAR VLHRELGWQV HLWTDDVSAL 51 RALCPDLPDV PFVHQDIHVR TWHSDAADID TAPVPDAVIE TFACDLPENV 101 LNIIRRHKPL WLNWEYLSAE ESNERLHLMP SPQEGVQKYF WFMGFSEKSG 151 GLIRERDYRE AVRFDTEALR RRLVLPEKNA PEWLLFGYRG DVWAKWLDMW 201 QQAGSLMTLL LAGAQIIDSL KQSGVIPQNA LQNEGGVFQT ASVRLVKIPF 251 VPQQDFDKLL HLADCAVIRG EDSFVRTQLA GKPFFWHIYP QDENVHLDKL 301 HAFWDKAYGF YTPETASVHR LLSDDLNGGE ALSATQRLEC WQTLQQHQNG 351 WRQGAEDWSR YLFGQPSASE KLAAFVSKHQ KIR* ORF32ng-l and ORF32-1 show 93.5% identity in 383 aa overlap: 10 20 30 40 50 59 orf 32-1 . pep MNTPPF-VCWIFCKVIDNFGDIGVSWRLARVLHRELGWQVHLWTDDVSALRALCPDLPDV III I II I II II I I I I II I I II II I I I I I II II II I I I I I I I II I I I I I II II II I I I orf 32ng-l MNTYAFPVCWIFCKVIDNFGDIGVSWRLARVLHRELGWQVHLWTDDVSALRALCPDLPDV 10 20 30 40 50 60 60 70 80 90 100 110 119 orf 32-1 . pep PCVHQDIHVRTWHSDAADIDTAPVPDWIETFACDLPENVLHIIRRHKPLWLNWEYLSAE I I I I I I II I I I I I I II I I II I II I I : I I I I I II I I II I I I: I II II II I II I I I I I II I orf32ng-l PFVHQDIHVRTWHSDAADIDTAPVPDAVIETFACDLPENVLNIIRRHKPLWLNWEYLSAE 70 80 90 100 110 120 120 130 140 150 160 170 179 orf32-1. pep ESNERLHLMPSPQEGVQKYFWFMGFSEKSGGLIRERDYCEAVRFDTEALRERLMLPEKNA II II II II I I I II II I I I I II I I II I II II II II II II I I II I I I II II : I I : II II I I orf32ng-l ESNERLHLMPSPQEGVQKYFWFMGFSEKSGGLIRERDYREAVRFDTEALRRRLVLPEKNA 130 140 150 160 170 180 180 190 200 210 220 230 239 orf32-1. pep SEWLLFGYRSDVWAKWLEMWRQAGSPMTLLLAGTQIIDSLKQSGVIPQDALQNDGDVFQT I II II II I : I II I I II : I I : II I I I I I I I I I : II II II I I II I I I I : I I I I : I I I II orf32ng-l PEWLLFGYRGDVWAKWLDMWQQAGSLMTLLLAGAQIIDSLKQSGVIPQNALQNEGGVFQT 190 200 210 220 230 240 240 250 260 270 280 290 299 orf32-1. pep ASVRLVKIPFVPQQDFDQLLHLADCAVIRGEDSFVRAQLAGKPFFWHIYPQDENVHLDKL II II II II I II II II II : I I I II I II I II I I II II I : II I I II I II I I I I II I I II II I I orf32ng-l ASVRLVKIPFVPQQDFDKLLHLADCAVIRGEDSFVRTQLAGKPFFWHIYPQDENVHLDKL 250 260 270 280 290 300 300 310 320 330 340 350 359 orf32-1. pep HAFWDKAHGFYTPETVSAHRRLSDDLNGGEALSATQRLECWQTLQQHQNGWRQGAEDWSR II II II I : I II II I I : I : II II I I I II II I I II II I I I I I I II I II I II II I II II I II orf32ng-l HAFWDKAYGFYTPETASVHRLLSDDLNGGEALSATQRLECWQTLQQHQNGWRQGAEDWSR 310 320 330 340 350 360 360 370 380 orf32-1. pep YLFGQPSAPEKLAAFVSKHQKIRX I I II II I I II II I I I II II I I II orf32ng-l YLFGQPSASEKLAAFVSKHQKIRX 370 380 On this basis, including the RGD sequence in the gonococcal protein, characteristic of adhesins, it is predicted that the proteins from N. meningitidis .and N.gonorrhoeae, and their epitopes, could be useful antigens for vaccines or diagnostics, or for raising antibodies. ORF32-1 (42kDa) was cloned in pET and pGex vectors and expressed in E.coli, as described above. The products of protein expression and purification were analyzed by SDS-PAGE. Figure 7 A shows the results of affinity purification of the His-fusion protein, and Figure 7B shows the results of expression of the GST-fusion in E.coli. Purified His-fusion protein was used to immunise mice, whose sera were used for ELISA, giving a positive result. These experiments confirm that ORF32-1 is a surface-exposed protein, and that it is a useful immunogen. Example 24 The following partial DNA sequence was identified in N. meningitidis <SEQ ID 197>: 1 .. TTGTTCCTGC GTGTNAAAGT GGGGCGTTTT TTCAGCAGTC CGGCGACGTG 51 GTTTCGGGNC AAAGACCCTG TAAATCAGGC GGTGTTGCGG CTGTATNCGG 101 ACGAGTGGCG GCA.ACTTCG GTACGTTGGA AAATAGNCGC AACGTCGCAC 151 AGCCTGTGGC TCTGCACGCT GCTCGGAATG CTGGTGTCGG TATTGTTGCT 201 GCTTTTGGTG CGGCAATATA CGTTCAACTG GGAAAGCACG CTGTTGAGCA 251 ATGCCGCTTC GGTACGCGCG GTGGAAATGT TGGCATGGCT GCCGTCGAAA 301 CTCGGTTTCC CTGTCCCCGA TGCGCGGTCG GTCATCGAAG GCCGTCTGAA 351 CGGCAATATT GCCGATGCGC GGGCTTGGTC GGGGCTGCTG GTCGNCAGTA 401 TCGCCTGCTA NGGCATCCTG CCGCGCCTG.. This corresponds to the -amino acid sequence <SEQ ID 198; ORF33>: 1 ..LFLRVKVGRF FSSPATWFRX KDPVNQAVLR LYXDEWRXTS VRWKIXATSH 51 SLWLCTLLGM LVSVLLLLLV RQYTFNWEST LLSNAASVRA VEMLAWLPSK 101 LGFPVPDARS VIEGRLNGNI ADARAWSGLL VXSIACXGIL PRL .. Further work revealed the complete nucleotide sequence <SEQ ID 199>: 1 ATGTTGAATC CATCCCGAAA ACTGGTTGAG CTGGTCCGTA TTTTGGACGA 51 AGGCGGTTTT ATTTTCAGCG GCGATCCCGT ACAGGCGACG GAGGCTTTGC 101 GCCGCGTGGA CGGCAGTACG GAGGAAAAAA TCATCCGTCG GGCGGAGATG 151 ATTGACAGGA ACCGTATGCT GCGGGAGACG TTGGAACGTG TGCGTGCGGG 201 GTCGTTCTGG TTGTGGGTGG TGGCGGCGAC GTTTGCATTT TTTACCGGTT 251 TTTCAGTCAC TTATCTTCTA ATGGACAATC AGGGTCTGAA TTTCTTTTTG 301 GTTTTGGCGG GCGTGTTGGG CATGAATACG CTGATGCTGG CAGTATGGTT 351 GGCAATGTTG TTCCTGCGTG TGAAAGTGGG GCGTTTTTTC AGCAGTCCGG 401 CGACGTGGTT TCGGGGCAAA GACCCTGTAA ATCAGGCGGT GTTGCGGCTG 451 TATGCGGACG AGTGGCGGCA ACCTTCGGTA CGTTGGAAAA TAGGCGCAAC 501 GTCGCACAGC CTGTGGCTCT GCACGCTGCT CGGAATGCTG GTGTCGGTAT 551 TGTTGCTGCT TTTGGTGCGG CAATATACGT TCAACTGGGA AAGCACGCTG 601 TTGAGCAATG CCGCTTCGGT ACGCGCGGTG GAAATGTTGG CATGGCTGCC 651 GTCGAAACTC GGTTTCCCTG TCCCCGATGC GCGGGCGGTC ATCGAAGGCC 701 GTCTGAACGG CAATATTGCC GATGCGCGGG CTTGGTCGGG GCTGCTGGTC 751 GGCAGTATCG CCTGCTACGG CATCCTGCCG CGCCTGCTGG CTTGGGTAGT 801 GTGTAAAATC CTTTTGAAAA CAAGCGAAAA CGGATTGGAT TTGGAAAAGC 851 CCTATTATCA GGCGGTCATC CGCCGCTGGC AGAACAAAAT CACCGATGCG 901 GATACGCGTC GGGAAACCGT GTCCGCCGTT TCACCGAAAA TCATCTTGAA 951 CGATGCGCCG AAATGGGCGG TCATGCTGGA GACCGAGTGG CAGGACGGCG 1001 AATGGTTCGA GGGCAGGCTG GCGCAGGAAT GGCTGGATAA GGGCGTTGCC 1051 ACCAATCGGG AACAGGTTGC CGCGCTGGAG ACAGAGCTGA AGCAGAAACC 1101 GGCGCAACTG CTTATCGGCG TGCGCGCCCA AACTGTGCCG GACCGCGGCG 1151 TGTTGCGGCA GATTGTCCGA CTCTCGGAAG CGGCGCAGGG CGGCGCGGTG 1201 GTGCAGCTTT TGGCGGAACA GGGGCTTTCA GACGACCTTT CGGAAAAGCT 1251 GGAACATTGG CGTAACGCGC TGGCCGAATG CGGCGCGGCG TGGCTTGAGC 1301 CTGACAGGGC GGCGCAGGAA GGGCGTTTGA AAGACCAATA A This corresponds to the amino acid sequence <SEQ LD 200; ORF33-l>: 1 MLNPSRKLVE LVRILDEGGF IFSGDPVQAT EALRRVDGST EEKIIRRAEM 51 IDRNRMLRET LERVRAGSFW LWWAATFAF FTGFSVTYLL MDNQGLNFFL 101 VLAGVLGMNT LMLAVWLAML FLRVKVGRFF SSPATWFRGK DPVNQAVLRL 151 YADEWRQPSV RWKIGATSHS LWLCTLLGML VSVLLLLLVR QYTFNWESTL 201 LSNAASVRAV EMLAWLPSKL GFPVPDARAV IEGRLNGNIA DARAWSGLLV 251 GSIACYGILP RLLAWWCKI LLKTSENGLD LEKPYYQAVI RRWQNKITDA 301 DTRRETVSAV SPKIILNDAP KWAVMLETEW QDGEWFEGRL AQEWLDKGVA 351 TNREQVAALE TELKQKPAQL LIGVRAQTVP DRGVLRQIVR LSEAAQGGAV 401 VQLLAEQGLS DDLSEKLEHW RNALAECGAA WLEPDRAAQE GRLKDQ* Computer analysis of this amino acid sequence gave the following results: Homology with a predicted ORF from N.meninsitidis (strain A) ORF33 shows 90.9% identity over a 143aa overlap with an ORF (ORF33a) from strain A of N meningitidis: 10 20 30 orf33 . pep LFLRVKVGRFFSSPATWFRXKDPVΝQAVLR I I I I I I I I I I I I I I I I I I I I I I I I I I I I I orf33a LMDΝQGLΝFFLVLAGVXGMΝTLMLAVWLAMLFLRVKVGRFFSSPATWFRGKDPVΝQAVLR 90 100 110 120 130 140 40 50 60 70 80 90 orf 33 . pep LYXDEWRXTSVRWKIXATSHSLWLCTLLGMLVSVLLLLLVRQYTFΝWESTLLSΝAASVRA II I I II I I II II I I II I I I I II I I II I I I II I II I II I I I II I II I II I:::: I I I orf 33a LYADEWRXPSVRWKIGATSHSLWLCTLLGMLVSVLLLLLVRQYTFΝWESTLLGDSSSVRL 150 160 170 180 190 200 100 110 120 130 140 orf 33. pep VEMLAWLPSKLGFPVPDARSVIEGRLΝGΝIADARAWSGLLVXSIACXGILPRL I I I I II I I : I II II I I I II : II I I I II I I I II I II I II I I I I I I I I I II II orf 33a VEMLAWLPAKLGFPVPDARAVIEGRLΝGΝIADARAWSGLLVGSIACYGILPRLLAWAVCK 210 220 230 240 250 260 orf33a ILXXTSEΝGLDLEKXXXXXXIRRWQΝKITDADTRRETVSAVSPKIVLΝDAPKWAVMLETE 270 280 290 300 310 320 The complete length ORF33a nucleotide sequence <SEQ ID 201> is: 1 ATGTTGAATC CATCCCGAAA ACTGGTTGAG CTGGTCCGTA TTTTGGAAGA 51 AGGCGGCTTT ATTTTCAGCG GCGATCCCGT GCAGGCGACG GAGGCTTTGC 101 GCCGCGTGGA CGGCAGTACG GAGGAAAAAA TCATCCGTCG GGCGAAGATG 151 ATCGACAGGA ACCGTATGCT GCGGGAGACG TTGGAACGTG TGCGTGCGGG 201 GTCGTTCTGG TTGTGGGTGG CGGCGGCGAC GTTTGCGTTT NTTACCGNTT 251 TTTCAGTTAC TTATCTTCTA ATGGACAATC AGGGTCTGAA TTTCTTTTTG 301 GTTTTGGCGG GCGTGNTGGG CATGAATACG CTGATGCTGG CAGTATGGTT 351 GGCAATGTTG TTCCTGCGCG TGAAAGTGGG GCGTTTTTTC AGCAGTCCGG 401 CGACGTGGTT TCGGGGCAAA GACCCTGTCA ATCAGGCGGT GTTGCGGCTG 451 TATGCGGACG AGTGGCGGCN ACCTTCGGTA CGTTGGAAAA TAGGCGCAAC 501 GTCGCACAGC CTGTGGCTCT GCACGCTGCT CGGAATGCTG GTGTCGGTAT 551 TGTTGCTGCT TTTGGTGCGG CAATATACGT TCAACTGGGA AAGCACGCTG 601 TTGGGCGATT CGTCTTCGGT ACGGCTGGTG GAAATGTTGG CATGGCTGCC 651 TGCGAAACTG GGTTTTCCCG TGCCTGATGC GCGGGCGGTC ATCGAAGGTC 701 GTCTGAACGG CAATATTGCC GATGCGCGGG CTTGGTCGGG GCTGCTGGTC 751 GGCAGTATCG CCTGCTACGG CATCCTGCCG CGCCTCTTGG CTTGGGCGGT 801 ATGCAAAATC CTTNTGNAAA CAAGCGAAAA CGGCTTGGAT TTGGAAAAGC 851 NCNNNNNTCN NNCGNTCATC CGCCGCTGGC AGAACAAAAT CACCGATGCG 901 GATACGCGTC GGGAAACCGT GTCCGCCGTT TCGCCGAAAA TCGTCTTGAA 951 CGATGCGCCG AAATGGGCGG TCATGCTGGA GACCGAATGG CAGGACGGCG 1001 AATGGTTCGA GGGCAGGCTG GCGCAGGAAT GGCTGGATAA GGGCGTTGCC 1051 GCCAATCGGG AACAGGTTGC CGCGCTGGAG ACAGAGCTGA AGCAGAAACC 1101 GGCGCAACTG CTTATCGGCG TGCGCGCCCA AACTGTGCCC GACCGCGGCG 1151 TGTTGCGGCA GATCGTCCGA CTTTCGGAAG CGGCGCAGGG CGGCGCGGTG 1201 GTGCANCTTT TGGCGGAACA GGGGCTTTCA GACGACCTTT CGGAAAAGCT 1251 GGAACATTGG CGTAACGCGC TGACCGAATG CGGCGCGGCG TGGCTGGAAC 1301 CCGACAGAGC GGCGCAGGAA GGCCGTCTGA AAACCAACGA CCGCACTTGA This encodes a protein having amino acid sequence <SEQ ID 202>: i MLNPSRKLVE LVRILEEGGF IFSGDPVQAT EALRRVDGST EEKIIRRAKM 51 IDRNRMLRET LERVRAGSFW LWVAAATFAF XTXFSVTYLL MDNQGLNFFL 101 VLAGVXGMNT LMLAVWLAML FLRVKVGRFF SSPATWFRGK DPVNQAVLRL 151 YADEWRXPSV RWKIGATSHS LWLCTLLGML VSVLLLLLVR QYTFNWESTL 201 LGDSSSVRLV EMLAWLPAKL GFPVPDARAV IEGRLNGNIA DARAWSGLLV 251 GSIACYGILP RLLAWAVCKI LXXTSENGLD LEKXXXXXXI RRWQNKITDA 301 DTRRETVSAV SPKIVLNDAP KWAVMLETEW QDGEWFEGRL AQEWLDKGVA 351 ANREQVAALE TELKQKPAQL LIGVRAQTVP DRGVLRQIVR LSEAAQGGAV 401 VXLLAEQGLS DDLSEKLEHW RNALTECGAA WLEPDRAAQE GRLKTNDRT* ORF33a and ORF33-1 show 94.1% identity in 444 aa overlap: 10 20 30 40 50 60 orf33a. pep MLNPSRKLVELVRILEEGGFIFSGDPVQATEALRRVDGSTEEKIIRRAKMIDRNRMLRET I I I I I I II I I II I I I : I I I I I I I II I II I II I I I II I II II I II II I I : II I I I I I I I II orf33-1 MLNPSRKLVELVRILDEGGFIFSGDPVQATEALRRVDGSTEEKIIRRAEMIDRNRMLRET 10 20 30 40 50 60 70 80 90 100 110 120 orf33a . pep LERVRAGSFWLWVAAATFAFXTXFSVTYLLMDNQGLNFFLVLAGVXGMNTLMLAVWLAML I II I I I I II I II M I I I I II I I I I I II I I II II I I I II I II II I I I I I I II I I II I I orf33-1 LERVRAGSFWLWWAATFAFFTGFSVTYLLMDNQGLNFFLVLAGVLGMNTLMLAVWLAML 70 80 90 100 110 120 130 140 150 160 170 180 orf33a . pep FLRVKVGRFFSSPATWFRGKDPVNQAVLRLYADEWRXPSVRWKIGATΞHSLWLCTLLGML I II II I I II I II I I I I I I II I II I I II II I I II II I I II I II II II I I II I I I I I I II I orf33-1 FLRVKVGRFFSSPATWFRGKDPVNQAVLRLYADEWRQPSVRWKIGATSHSLWLCTLLGML 130 140 150 160 170 180 190 200 210 220 230 240 orf33a . pep VSVLLLLLVRQYTFNWESTLLGDSSSVRLVEMLAWLPAKLGFPVPDARAVIEGRLNGNIA I II I I I I II II I II II II I I I :::: I II I II I I II I : I I I I II I I I I I I II I I I II I I I orf33-1 VSVLLLLLVRQYTFNWESTLLSNAASVRAVEMLAWLPSKLGFPVPDARAVIEGRLNGNIA 190 200 210 220 230 240 250 260 270 280 290 300 orf33a . pep DARAWSGLLVGSIACYGILPRLLAWAVCKILXXTSENGLDLEKXXXXXXIRRWQNKITDA I I II II I I II I I I I I II I I I I II I I: I II I I II I I I II I II II I I I I II I II orf33-l DARAWSGLLVGSIACYGILPRLLAWWCKILLKTSENGLDLEKPYYQAVIRRWQNKITDA 250 260 270 280 290 300 310 320 330 340 350 360 orf33a . pep DTRRETVSAVSPKIVLNDAPKWAVMLETEWQDGEWFEGRLAQEWLDKGVAANREQVAALE I II I I I II I II I I I : I I II II II I I I II II I II I II II II I I II II II I I : I I I II I I II orf33-1 DTRRETVSAVSPKIILNDAPKWAVMLETEWQDGEWFEGRLAQEWLDKGVATNREQVAALE 310 320 330 340 350 360 370 380 390 400 410 420 orf33a. pep TELKQKPAQLLIGVRAQTVPDRGVLRQIVRLSEAAQGGAWXLLAEQGLSDDLSEKLEHW II II I II I I I I I II I I II I I II I II II I I I II I II II II I I I I I I II II I I I II II I II orf33-1 TELKQKPAQLLIGVRAQTVPDRGVLRQIVRLSEAAQGGAWQLLAEQGLSDDLSEKLEHW 370 380 390 400 410 420 430 440 450 orf 33a . pep RNALTECGAAWLEPDRAAQEGRLKTNDRTX I I I I : I I I I I I I I I I I I I I I I I I I orf33-l RNALAECGAAWLEPDRAAQEGRLKDQX 430 440 Homology with a predicted ORF from N.sonorrhoeae ORF33 shows 91.6% identity over a 143aa overlap with a predicted ORF (ORF33.ng) from N. gonorrhoeae: orf33. pep LFLRVKVGRFFSSPATWFRXKDPVNQAVLR 30 I II I II I I II I I II I I I I I I II I I I II I orf33ng LMDNQGLNFFLVLAGVLGMNTLMLAVWLATLFLRVKVGRFFSSPATWFRGKGPVNQAVLR 100 orf33.pep LYXDEWRXTSVRWKIXATSHSLWLCTLLGMLVSVLLLLLVRQYTFNWESTLLSNAASVRA 90 M M M M M I I : I I I I II I I I II II I I II II I I I I II I II I II II II I I I II I orf33ng LYADQWRQPSVRWKIGATAHSLWLCTLLGMLVSVLLLLLVRQYTFNWESTLLSNAASVRA 160 or 33.pep VEMLAWLPSKLGFPVPDARSVIEGRLNGNIADARAWSGLLVXSIACXGILPRL 143 I II I II I II II I II II I IM I I II I I II I I II I II I I I II I MM I I I I II orf33ng VEMLAWLPSKLGFPVPDARAVIEGRLNGNIADARAWSGLLVGSIVCYGILPRLLAWWCK 220 An ORF33ng nucleotide sequence <SEQ ID 203> was predicted to encode a protein having amino acid sequence <SEQ ID 204>: 1 MIDRDRMLRD TLERVRAGSF WLWVWASMM FTAGFSGTYL LMDNQGLNFF 51 LVLAGVLGMN TLMLAVWLAT LFLRVKVGRF FSSPATWFRG KGPVNQAVLR 101 LYADQWRQPS VRWKIGATAH SLWLCTLLGM LVSVLLLLLV RQYTFNWEST 151 LLSNAASVRA VEMLAWLPSK LGFPVPDARA VIEGRLNGNI ADARAWSGLL 201 VGSIVCYGIL PRLLAWWCK ILLKTSENGL DLEKTYYQAV IRRWQNKITD 251 ADTRRETVSA VSPKIVLNDA PKWALMLETE WQDGQWFEGR LAQEWLDKGV 301 AANREQVAAL ETELKQKPAQ LLIGVRAQTV PDRGVLRQIV RLSEAAQGGA 351 WQLLAEQGL SDDLSEKLEH WRNALTECGA AWLEPDRVAQ EGRLKDQ* Further sequence analysis revealed the following DNA sequence <SEQ ID 205>: 1 ATGTTGaatC CATCCCgaAA ACTGgttgag ctGgTCCgtA Ttttgaataa 51 agggggtTTT attttcagcg gcgatcctgt gcaggcgacg gaggctttgc 101 gccgcgtgga cggcAGTACG GAggAaaaaa tcttccgtcg GGCGGAGAtg 151 atcgACAGGg accgtatgtt gcgggACaCg TtggaacGTG TGCGTGCggg 201 gtcgtTctgG TTATGGGTGG TggtggCAtC gATGATGTtt aCCGCCGGAT 251 TTTCAGgcac ttatCttCTG ATGGACaatC AGGGGCtGAA TtTCTTTTTA 301 GTTTTggcgG GAGTGTtggG CATGaatacG ctgATGCTGG CAGTATGGtt 351 gGCAACGTTG TTCCTGCGCG TGAAAGTGGG ACGGTTTTTC AGCAGTCCGG 401 CGACGTGGTT TCGGGGCAAA GGCCCTGTAA ATCAGGCGGT GTTGCGGCTG 451 TATGCGGACC AGTGGCGGCA ACCTTCGGTA CGATGGAAAA TAGGCGCAAC 501 GGCGCACAGC TTGTGGCTCT GCACGCTGCT CGGAATGCTG GTGTCGGTAT 551 TGCTGCTGCT TTTGGTGCGG CAATATACGT TCAACTGGGA AAGCACGCTG 601 TTGAGCAATG CCGCTTCGGT ACGCGCGGTG GAAATGTTGG CATGGCTGCC 651 GTCGAAACTC GGTTTCCCTG TCCCCGATGC GCGGGCGGTC ATCGAAGGTC 701 GTCTGAACGG CAATATTGCC GATGCGCGGG CTTGGTCGGG GCTGCTGGTC 751 GGCAGTATCG TCTGCTACGG CATCCTGCCG CGCCTCTTGG CTTGGGTAGT 801 GTGTAAAATC CTTTTGAAAA CAAGCGAAAA CGGattgGAT TTGGAAAAAA 851 CCTATTATCA GGCGGTCATC CGCCGCTGGC AGAACAAAAT CACCGATGCG 901 GATACGCGTC GGGAAACCGT GTCCGCCGTT TCGCcgaAAA TCGTCTTGAA 951 CGATGCGCCG AAATGGGCGC TCATGCTGGA GACCGAGTGG CAGGACGGCC 1001 AATGGTTCGA GGGCAGGCTG GCGCAGGAAT GGCTGGATAA GGGCGTTGCC 1051 GCCAATCGGG AACAGGTTGC CGCGCTGGAG ACAGAGCTGA AGCAGAAACC 1101 GGCGCAACTG CTTATCGGCG TACGCGCCCA AACTGTGCCG GACCGGGGCG 1151 TGCTGCGGCA GATTGTGCGG CTTTCGGAAG CGGCGCAGGG CGGCGCGGTG 1201 GTGCAGCTTT TGGCGGAACA GGGGCTTTCA GACGACCTTT CGGAAAAGCT 1251 GGAACATTGG CGTAACGCGC TGACCGAATG CGGCGCGGCG TGGCTTGAGC 1301 CTGACAGGGT GGCGCAGGAA GGCCGTTTGA AAGACCAATA A This encodes a protein having amino acid sequence <SEQ ID 206; ORF33ng-l>: 1 MLNPSRKLVE LVRILNKGGF IFSGDPVQAT EALRRVDGST EEKIFRRAEM 51 IDRDRMLRDT LERVRAGSFW LWVWASMMF TAGFSGTYLL MDNQGLNFFL 101 VLAGVLGMNT LMLAVWLATL FLRVKVGRFF SSPATWFRGK GPVNQAVLRL 151 YADQWRQPSV RWKIGATAHS LWLCTLLGML VSVLLLLLVR QYTFNWESTL 201 LSNAASVRAV EMLAWLPSKL GFPVPDARAV IEGRLNGNIA DARAWSGLLV 251 GSIVCYGILP RLLAWWCKI LLKTSENGLD LEKTYYQAVI RRWQNKITDA 301 DTRRETVSAV SPKIVLNDAP KWALMLETEW QDGQWFEGRL AQEWLDKGVA 351 ANREQVAALE TELKQKPAQL LIGVRAQTVP DRGVLRQIVR LSEAAQGGAV 401 VQLLAEQGLS DDLSEKLEHW RNALTECGAA WLEPDRVAQE GRLKDQ* ORF33ng-l and ORF33-1 show 94.6% identity in 446 aa overlap: 10 20 30 40 50 60 orf33-1.pep MLNPSRKLVELVRILDEGGFIFSGDPVQATEALRRVDGSTEEKIIRRAEMIDRNRMLRET I I I I II II II I I II I :: II I II II I I I I I II I II II II II I I II: I II II II I: II I I: I orf33ng-l MLNPSRKLVELVRILNKGGFIFSGDPVQATEALRRVDGSTEEKIFRRAEMIDRDRMLRDT 10 20 30 40 50 60 70 80 90 100 110 120 orf33-l .pep LERVRAGSFWLWWAATFAFFTGFSVTYLLMDNQGLNFFLVLAGVLGMNTLMLAVWLAML II I II I I I I II II I : I :: I M II I I I I I II I I I I II II II I I I I II I I II I I I I orf33ng-l LERVRAGSFWLWVWASMMFTAGFSGTYLLMDNQGLNFFLVLAGVLGMNTLMLAVWLATL 70 80 90 100 110 120 130 140 150 160 170 180 orf33-l .pep FLRVKVGRFFSSPATWFRGKDPVNQAVLRLYADEWRQPSVRWKIGATSHSLWLCTLLGML I II I II II I I I I II II I I II I I II I I II II I I : I II II I I I I I II I : I I I I II I I I II I orf33ng-l FLRVKVGRFFSSPATWFRGKGPVNQAVLRLYADQWRQPSVRWKIGATAHSLWLCTLLGML 130 140 150 160 170 180 190 200 210 220 230 240 orf33-l .pep VSVLLLLLVRQYTFNWESTLLSNAASVRAVEMLAWLPSKLGFPVPDARAVIEGRLNGNIA I I II II II II II II I I II I II II I II II II I II II I I I I I I II II I I I I I I I I II II I II orf33ng-l VSVLLLLLVRQYTFNWESTLLSNAASVRAVEMLAWLPSKLGFPVPDARAVIEGRLNGNIA 190 200 210 220 230 240 250 260 270 280 290 300 orf33-l .pep DARAWSGLLVGSIACYGILPRLLAWWCKILLKTSENGLDLEKPYYQAVIRRWQNKITDA II I I I II I II I II: I II II I II I II I I II II I II II II I I II I I I I II I I I II I I I II I orf33ng-l DARAWSGLLVGSIVCYGILPRLLAWWCKILLKTSENGLDLEKTYYQAVIRRWQNKITDA 250 260 270 280 290 300 310 320 330 340 350 360 orf33-1. pep DTRRETVSAVSPKIILNDAPKWAVMLETEWQDGEWFEGRLAQEWLDKGVATNREQVAALE II II I II II I I II I : I I II I I I I : I II II I II I : I II II I II I I I II I I I M II I I I I II orf33ng-l DTRRETVSAVSPKIVLNDAPKWALMLETEWQDGQWFEGRLAQEWLDKGVAANREQVAALE 310 320 330 340 350 360 370 380 390 400 410 420 orf33-1. pep TELKQKPAQLLIGVRAQTVPDRGVLRQIVRLSEAAQGGAWQLLAEQGLSDDLSEKLEHW I I II I II I II I II II II II I II I I II II I I I II II I I I I I I I I II I I I II II I I II II I I orf33ng-l TELKQKPAQLLIGVRAQTVPDRGVLRQIVRLSEAAQGGAWQLLAEQGLSDDLSEKLEHW 370 380 390 400 410 420 430 440 orf33-1. pep RNALAECGAAWLEPDRAAQEGRLKDQX I II I: I II II I II II I: I I II II I II I orf33ng-l RNALTECGAAWLEPDRVAQEGRLKDQX 430 440 Based on the presence of several putative transmembrane domains in the gonococcal protein, it is predicted that the proteins from N. meningitidis and N.gonorrhoeae, and their epitopes, could be useful antigens for vaccines or diagnostics, or for raising antibodies. Example 25 The following partial DNA sequence was identified in N. meningitidis <SEQ ID 207>: 1 .. CAGAAGAGTT TGTCGAGAAT TTCTTTATGG GGTTTGGGCG GCGTGTTTTT 51 CGGGGTGTCC GGTCTGGTAT GGTTTTCTTT GGGCGTTTCT TT.GAGTGCG 101 CCTGTTTTTC GGGTGTTTCT TTTCGGGGTT CGGGACGGGG GACGTTTGTG 151 GGCAGTACGG GGGTTTCTTT GAGTGTGTTT TCAGCTTGTG TTCC . GGCGT 201 CGTCCGGCTG CCTGTCGGTT TGAGCTGTGT CGGCAGGTTG CG..GTTTGA 251 CCCGGTTTTT CTTGGGTGCG GCAGGGGACG TCATTCTCCT GCCGCTTTCG 301 TCTGTGCCGT CCGGCTGTGC GGGTTCGGAT GAGGCGGCGT GGTGGTGTTC 351 GGGTTGGGCG GCATCTTGTI CCGACTACGC CGTTTGGCAG CCAGAATTCG 401 GTTTCGCGGG GGCTGTCGGT GTGTTGCGGT TCGGCTTGAA GGGTTTTGTC 451 GTCC. This corresponds to the .amino acid sequence <SEQ ID 208; ORF34>: 1 ..QKSLSRISLW GLGGVFFGVS GLVWFSLGVS XECACFSGVS FRGSGRGTFV 51 GSTGVSLSVF SACVXGWRL PVGLSCVGRL XXLTRFFLGA AGDVILLPLS 101 SVPSGCAGSD EAAWWCSGWA ASCPTTPFGS QNSVSRGLSV CCGSA*RVLS 151 S.. Further work revealed the complete nucleotide sequence <SEQ ID 209>: 1 ATGATGATGC CGTTCATAAT GCTTCCTTGG ATTGCkGGTG TGCCTGCCGT 51 GCCGGGTCAG AATAGGTTGT CCAGAATTTC TTTATGGGGT TTGGGCGGCG 101 TGTTTTTCGG GGTGTCCGGT TTGGTATGGT TTTCTTTGGG CGTTTCTTTG 151 GGCTGCGCCT GTTTTTCGGG TGTTTCTTTT CGGGGTTCGG GACGGGGGAC 201 GTTTGTGGGC AGTACGGGGG TTTCTTTGAG TGTGTTTTCA GCTTGTGTTC 251 CGGCGTCGTC CGGCTGCCTG TCGGTTTGAG CTGTGTCGGC AGGTTGCGGT 301 TTGACCCGGT TTTTCTTGGG TGCGGCAGGG GACGGCAGTC CGCTGCCGCT 351 TTCGTCTGTG CCGTCCGGCT GTGCGGGTTC GGATGAGGCG GCGTGGTGGT 401 GTTCGGGTTG GGCGGCATCT TGTCCGACTA CGCCGTTTGG CAGCCAGAAT 451 TCGGTTTCGC GGGGGCTGTC GGTGTGTTGC GGTTCGGCTT GAAGGGTTTT 501 GTCGCCGTTC GGGTTGAATG TGCTGACGAT GCCTATTGCC AATGCGCCGA 551 TGGCGGCGAT ACAGATGAGC AATACGGCGC GTATCAGGAG TTTGGGGGTC 601 AGCCTGAAGG GTTTGTTCGG TTTTTTTGCC ATTTTGATTG TGCTTTTGGG 651 GTGTCGGGCA ATGCCGTCTG AAGGCGGTTC AGACGGCATT GCCGAGTCAG 701 CGTTGGACGT AGTTTTGGTA GAGGGTGATG ACTTTTTGTA CGCCGACGGT 751 GGTGCTGACT TTTTGGGTAA TCTGCGCCTG TTCTTCGGGG GTGAGGATGC 801 CCATAACGTA GGTTACGTTG CCGTAGGTAA CGATTTTGAC GCGCGCCTGT 851 GTGGCGGGGC TGATGCCCAA CAGCGTGGCG CGGACTTTGG ATGTGTTCCA 901 AGTGTCGCCG GCGATGTCGC CGGCAGTGCG CGGCAGGGAG GCGACGGTAA 951 TATAGTTGTA CACGCCTTCG GCGGCCTGTT CGGAACGTGC AATCTGACCG 1001 ACGAACTGTT TTTCGCCTTC GGTGGCGACT TGTCCGAGCA GCAGCAGGTG 1051 GCGGTTGTAG CCGACGACGG AGATTTGGGG CGTGTAGCCT TTGGTTTGGT 1101 TGTTTTGGCG CAGATAGGAA CGGGCGGTGG TTTCGATACG CAACGCCATA 1151 ACGTTGTCGT CGGTTTGCGC GCCGGTGGTT CGGCGGTCGA CGGCGGATTT 1201 CGCGCCGACG GCGGCGCTTC CGATTACTGC GCTGACGCAG CCGCTAAGGG 1251 CAAGGCTGAA AATGGCGGCA ATCAGGGTGC GGACGGTGTG CGGTTTGGGT 1301 TTCATCGGGT GCTTCCTTTC TTGGGCGTTT CAGACGGCAT TGCTTTGCGC 1351 CATGCCGTCT GA This corresponds to the amino acid sequence <SEQ ID 210; ORF34-l>: 1 MMMPFIMLPW IAGVPAVPGQ NRLSRISLWG LGGVFFGVSG LVWFSLGVSL 51 GCACFSGVSF RGSGRGTFVG STGVSLSVFS ACVPASSGCL SV*AVSAGCG 101 LTRFFLGAAG DGSPLPLSSV PSGCAGSDEA AWWCSGWAAS CPTTPFGSQN 151 SVSRGLSVCC GSA*RVLSPF GLNVLTMPIA NAPMAAIQMS NTARIRSLGV 201 SLKGLFGFFA ILIVLLGCRA MPSEGGSDGI AESALDWLV EGDDFLYADG 251 GADFLGNLRL FFGGEDAHNV GYVAVGNDFD ARLCGGADAQ QRGADFGCVP 301 SVAGDVAGSA RQGGDGNIW HAFGGLFGTC NLTDELFFAF GGDLSEQQQV 351 AWADDGDLG RVAFGLWLA QIGTGGGFDT QRHNVWGLR AGGSAVDGGF 401 RADGGASDYC ADAAAKGKAE NGGNQGADGV RFGFHRVLPF LGVSDGIALR 451 HAV* Computer analysis of this amino acid sequence gave the following results: Homology with a predicted ORF from N.meninsitidis (strain Al ORF34 shows 73.3% identity over a 161aa overlap with an ORF (ORF34a) from strain A of N meningitidis: 10 20 30 orf34.pep QKSLSRISLWGLGGVFFGVSGLVWFSLGVSXE- -CAC orf34a MMXPXIMLPWIAGVPAVPGQKRLSRXSLWGLGGXFFGVSGLVWFSLGVSXSLGVSXGCAC 10 20 30 40 50 60 40 50 60 70 80 90 orf34.pep FSGVSFRGSGRGTFVGSTGVSLSVFSACVXGWRLPVGLSCVGRLXX- -LTRFFLGA orf34a FSGVSFRGSGRGTFVGSTGVSLSVFSACA- —PASSGCLSVXAVSAGCGLTRXFXGA 70 80 90 100 110 100 110 120 130 140 150 orf34.pep AGDVILLPLSSVPSGCAGSDEAAWWCSGWAASCPTTPFGSQΝSVSRGLSVCCGSAXRVLS III II I I I I I II II I : I I I I I I I II I II I II I I I I II II II I I I I I II : I I II orf34a AGDGSPLPLSSVPSGCAGADEEAXXCSGWAASCPTTPFGSQΝSVSRGLSVCCGSVWRVLS 120 130 140 150 160 170 orf34.pep orf34a PFGXΝVLTMPIAΝAPMAVIQMSΝTARIRSLGVSLKGLFXFFAILIVLLGCRAMPSEGGSD 180 190 200 210 220 230 The complete length ORF34a nucleotide sequence <SEQ ID 211> is: 1 ATGATGATΝC CGTTΝATAAT GCTTCCTTGG ATTGCGGGTG TGCCTGCCGT 51 GCCGGGTCAG AAGAGGTTGT CGAGAAΝTTC TTTATGGGGT TTAGGCGGCΝ 101 TGTTTTTCGG GGTGTCCGGT TTGGTATGGT TTTCTTTGGG CGTTTCTΝTT 151 TCTTTGGGTG TTTCTΝTGGG CTGTGCCTGT TTTTCGGGTG TTTCTTTTCG 201 GGGTTCGGGA CGGGGGACGT TTGTGGGCAG TACΝGGGGTT TCTTTGAGTG 251 TGTTTTCAGC TTGTGCTCCG GCGTCGTCCG GCTGCCTGTC GGTTTΝAGCT 301 GTGTCGGCAG GTTGCGGTTT GACCCGGΝTT TTCTTΝGGTG CGGCAGGGGA 351 CGGCAGTCCG CTGCCGCTTT CGTCTGTGCC GTCCGGCTGT GCGGGTGCGG 401 ATGAGGAGGC GTΝGTΝGTGT TCGGGTTGGG CGGCATCTTG TCCGACTACG 451 CCGTTTGGCA GCCAGAATTC GGTTTCGCGG GGGCTGTCGG TGTGTTGCGG 501 TTCGGTΝTGG AGGGTTTTGT CΝCCGTTCGG GTΝGAATGTG CTGACGATGC 551 CTATTGCCAA TGCGCCGATG GCGGTGATAC AGATGAGCAA TACGGCGCGT 601 ATCAGGAGTT TGGGGGTCAG CCTGAAGGGT TTGTTCΝGTT TTTTTGCCAT 651 TTTGATTGTG CTTTTGGGGT GTCGGGCAAT GCCGTCTGAA GGCGGTTCAG 701 ACGGCATTGC CGAGTCAGCG TTGGACGTAG TTTΝGGTAGA GGGTGATGAC 751 TTTTTGTACG CCGACGGTGG TGCTGACTTT TTGGGTAATC TGCGCCTGTT 801 CTTCGGGGGT GAGGATGCCC ATAACGTAGG TTACGTTGCC GTAGGTAACG 851 ATTTTGACGC GCGCCTGTGT GGCGGGGCTG ATGCCCAACA GCGTGGCGCG 901 GACTTTGGAT GTGTTCCAAG TGTCGCCGGC GATGTCGCCG GCAGTGCGCG 951 GCAGGGAGGC GACGGTAATG TAΝTTGTACA CGCCTTCGGC GGCCTGTTCG 1001 GAACGTGCAA TCTGACCGAC GAACTGTTTC TCGCCTTCGG TGGCGACTTG 1051 TCCGAGCAGC AGCAGGTGGC GGTTGTAGCC GACAACGGAG ATTTGGGGCG 1101 TGTAΝCCTTT GGTTTGGTTG TTTTGGCGCA GATAGGAGCG GGCGGTGGTT 1151 TCGATACGCA GCGCCATTAC GTTGTCGTCG GTTΝGCGCGC CGGTGGTTCG 1201 GCGGTCGACG GCGGATTTCG CGCCGACCGC CGCGCCGCCG ACGACTGCGC 1251 TGACGCAGCC GCCGAGGGCA AGGCTGAGGA CGGCGGCAGT CAGGGTGCGG 1301 ACGGTGTGCG GTTTGGGTTT CATCGGGTGC TTCCTTTCTT GGGCGTTTCA 1351 GACGGCATTG CTTTGCGCCA TGCCGTCTGA This encodes a protein having amino acid sequence <SEQ ID 212>: 1 MMXPXIMLPW IAGVPAVPGQ KRLSRXSLWG LGGXFFGVSG LVWFSLGVSX 51 SLGVSXGCAC FSGVSFRGSG RGTFVGSTGV SLSVFSACAP ASSGCLSVXA 101 VSAGCGLTRX FXGAAGDGSP LPLSSVPSGC AGADEEAXXC SGWAASCPTT 151 PFGSQNSVSR GLSVCCGSVW RVLSPFGXNV LTMPIANAPM AVIQMSNTAR 201 IRSLGVSLKG LFXFFAILIV LLGCRAMPSE GGSDGIAESA LDWXVEGDD 251 FLYADGGADF LGNLRLFFGG EDAHNVGYVA VGNDFDARLC GGADAQQRGA 301 DFGCVPSVAG DVAGSARQGG DGNVXVHAFG GLFGTCNLTD ELFLAFGGDL 351 SEQQQVAWA DNGDLGRVXF GLWLAQIGA GGGFDTQRHY WVGXRAGGS 401 AVDGGFRADR RAADDCADAA AEGKAEDGGS QGADGVRFGF HRVLPFLGVS 451 DGIALRHAV* ORF34a and ORF34-1 show 91.3% identity in 459 aa overlap: 10 20 30 40 50 60 orf34a.pep MMXPXIMLPWIAGVPAVPGQKRLSRXSLWGLGGXFFGVSGLVWFSLGVSXSLGVSXGCAC I I I I II II II II II I I I I : I II I I I I II I I II II I II II I I I I II I II I orf34-1 MMMPFIMLPWIAGVPAVPGQNRLSRISLWGLGGVFFGVSGLVWFSLGVSL GCAC 10 20 30 40 50 70 80 90 100 110 120 orf34a.pep FSGVSFRGSGRGTFVGSTGVSLSVFSACAPASSGCLSVXAVSAGCGLTRXFXGAAGDGSP II II I II I I I I I I II I I I I II II I II I I : I I II II I I II I I I II II I I I I I I I I II I I orf34-1 FSGVSFRGSGRGTFVGSTGVSLSVFSACVPASSGCLSVXAVSAGCGLTRFFLGAAGDGSP 60 70 80 90 100 110 130 140 150 160 170 180 orf34a.pep LPLSSVPSGCAGADEEAXXCSGWAASCPTTPFGSQNSVSRGLSVCCGSVWRVLSPFGXNV I I II I II I II I I : I I I II II I I II II II II II II I I I II I I II I I : I I I I I I I II orf34-1 LPLSSVPSGCAGSDEAAWWCSGWAASCPTTPFGSQNSVSRGLSVCCGSAXRVLSPFGLNV 120 130 140 150 160 170 190 200 210 220 230 240 orf 34a. pep LTMPIANAPMAVIQMSNTARIRSLGVSLKGLFXFFAILIVLLGCRAMPSEGGSDGIAESA I I I I I I I I I I I : I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I orf 34-1 LTMPIANAPMAAIQMSNTARIRSLGVSLKGLFGFFAILIVLLGCRAMPSEGGSDGIAESA 180 190 200 210 220 230 250 260 270 280 290 300 orf 34a . pep LDWXVEGDDFLYADGGADFLGNLRLFFGGEDAHNVGYVAVGNDFDARLCGGADAQQRGA I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I orf 34-1 LDWLVEGDDFLYADGGADFLGNLRLFFGGEDAHNVGYVAVGNDFDARLCGGADAQQRGA 240 250 260 270 280 290 310 320 330 340 350 360 orf 34a . pep DFGCVPSVAGDVAGSARQGGDGNVXVHAFGGLFGTCNLTDELFLAFGGDLSEQQQVAWA I I I I I I I I I I I I I I I I I I I I I I I : I I I I I I I I I I I I I I I I I I : I I I I I I I I I I I I I I I I orf 34-1 DFGCVPSVAGDVAGSARQGGDGNIWHAFGGLFGTCNLTDELFFAFGGDLSEQQQVAWA 300 310 320 330 340 350 370 380 390 400 410 420 orf 34a . pep DNGDLGRVXFGLWLAQIGAGGGFDTQRHYVWGXRAGGSAVDGGFRADRRAADDCADAA I: I I I I I I I I I I I I I I I I : I I I I I I I I I I I I I I I I I I I I I I I I I I I I: I I I I I I orf 34-1 DDGDLGRVAFGLWLAQIGTGGGFDTQRHNVWGLRAGGSAVDGGFRADGGASDYCADAA 360 370 380 390 400 410 430 440 450 460 orf 34a. pep AEGKAEDGGSQGADGVRFGFHRVLPFLGVSDGIALRHAVX I : I I I I : I I : I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I orf34-1 AKGKAENGGNQGADGVRFGFHRVLPFLGVSDGIALRHAVX 420 430 440 450 Homology with a predicted ORF from N.sonorrhoeae ORF34 shows 77.6% identity over a 161aa overlap with a predicted ORF (ORF34.ng) from N gonorrhoeae: orf 34. pep QKSLSRISLWGLGGVFFGVSGLVWFSLGVSXE CAC 35 II I I I I I I I II : I II II II II II I II I I I III orf34ng MMMPFIMLPWIAGVPAVPGQKRLSRISLWGLAGVFFGVSGLVWFSLGVSFSLGVSLGCAC 60 orf34.pep FSGVSFRGSGRGTFVGSTGVSLSVFSACVXGWRLPVGLSCV GRLXXLTRFFLGA 90 II I II I I I II M II II II II I I M II I I : I I : I : II II I I II I I orf34ng FSGVSFRGSGWGAFVGSTGVSLSVFSACVP VPVNESAARAASEGR—GLTRFFLGA 114 orf34.pep AGDVILLPLSSVPSGCAGSDEAAWWCSGWAASCPTTPFGSQNSVSRGLSVCCGSAXRVLS 150 III I II I II II I I II II I I I II II I II I II I I : II I I II I I I I I I I II I II : I II I orf34ng AGDGSPLPLSSVPSGCAGSDEAAWWCSGWAASCPTAPFGSQNSVSRGLSVCCGSVWRVLS 174 orf34.pep S 175 orf34ng PFGLNVLTMPTANAPMAVIQMSNTARIRSLGVSLKGLFGFFAILIVLLGCRAMPSEGGSD 234 The complete length ORF34ng nucleotide sequence <SEQ ID 213> is: 1 ATGATGATGC CGTTCATAAT GCTTCCTTGG ATTGCGGGTG TGCCTGCCGT 51 GCCGGGTCAA AAGAGGTTGT CGAGAATCTC TTTATGGGGT TTGGCCGGCG 101 TGTTTTTCGG GGTGTCCGGT TTGGTATGGT TTTCTTTGGG CGTTTCTTTT 151 TCTTTGGGTG TTTCTTTGGG CTGCGCCTGT TTTTCGGGTG TTTCTTTTCG 201 GGGTTCGGGA TGGGGGGCGT TTGTGGGCAG TACGGGGGTT TCTTTGAGTG 251 TGTTTTCAGC TTGTGTTCCG GTGCCGGTTA ACGAATCGGC TGCCCGGGCC 301 GCATCCGAAG GGCGCGGTTT gACCCGGTTT TTCTTGGGTG CGGCAGGGGA 351 CGGCAGTCCG CTGCCGCTTT CTTCTGTGCC GTCCGGCTGT GCGGGTTCGG 401 ATGAGGCGGC GTGGTGGTGT TCGGGTTGGG CGGCATCTTG TCCGACGGCG 451 CCGTTTGGCA GCCAGAATTC GGTTTCGCGG GGGCTGTCGG TGTGTTGCGG 501 TTCGGTTTGG AGGGTTTTGT CGCCGTTCGG GTTGAATGTG CTGACGATGC 551 CTACTGCCAA TGCGCCGATG GCGGTGATAC AGATGAGCAA TACGGCGCGT 601 ATCAGGAGTT TGGGGGTCAG CCTGAAGGGT TTGTTCGGTT TTTTTGCCAT 651 TTTGATTGTG CTTTTGGGGT GTCGGGCAAT GCCGTCTGAA GGCGGTTCAG 701 ACGGCATTGC CGAGTCAGCG TTGGACGTAG TTTTGGTAGA GGGTAATGAC 751 TTTTTGTACG CCGAcggTGG TGCTGACTTT TTGGGTAATC TGCGCCTGTT 801 CTTCGGGGGT GAGGATGCCC ATAACGTAGG TTACATTGCC GTAGGTAATG 851 ATTTTGACGC GCGCCTGTGT AGCGGGGCTG ATGCCCAGCA GcgtgGCGCG 901 GACTTTGGAC GTGTTCCAAG TGTCGCCGGC GATGTCGCCC GCAGTGCGCG 951 GCAGGGAGGC GACGGTAATG TAGTTGTATA CGCCTTCGGC GGCCTGTTCG 1001 GAACGTGCAA TCTGACCGAC GAACTGTTTT TCGCCTTCGG TGGCGACTTG 1051 TCCGAGCAGC AGCAGGTGGC GGTTGTAGCC GACGACGGAG ATTTGGGGCG 1101 TGTAGCCTTT GGTTTGGTTG TTTTGGCGCA GGTAGGAACG GGCGGTGGTT 1151 TCGATACGCA ACGCCATAAC GTtgtCATCG GTTtgcgcgc CGGTGGTTcg 1201 gCGGTCGATG ACGGATTTTG CGCCGACGGC GGCCCCGCCG ACGACTGCGC 1251 TGAAGCAGCC GCCGAGGGCA AGGCTGAGGA CGGCGGCAAT CAGGGTGCGG 1301 ACGGTGTGTG GTTTGGGTTT CATCGGGGAC TTCCTTTCTT GGGCGTTTCA 1351 GACGGCATTG CTTTGCGCCA TGCCGTCTGA This encodes a protein having amino acid sequence <SEQ ID 214>: 1 MMMPFIMLPW IAGVPAVPGQ KRLSRISLWG LAGVFFGVSG LVWFSLGVΞF 51 SLGVSLGCAC FSGVSFRGSG WGAFVGSTGV SLSVFSACVP VPVNESAARA 101 ASEGRGLTRF FLGAAGDGSP LPLSSVPSGC AGSDEAAWWC SGWAASCPTA 151 PFGSQNSVSR GLSVCCGSVW RVLSPFGLNV LTMPTANAPM AVIQMSNTAR 201 IRSLGVSLKG LFGFFAILIV LLGCRAMPSE GGSDGIAESA LDWLVEGND 251 FLYADGGADF LGNLRLFFGG EDAHNVGYIA VGNDFDARLC ΞGADAQQRGA 301 DFGRVPSVAG DVARSARQGG DGNVWYAFG GLFGTCNLTD ELFFAFGGDL 351 SEQQQVAWA DDGDLGRVAF GLWLAQVGT GGGFDTQRHN WIGLRAGGS 401 AVDDGFCADG GPADDCAEAA AEGKAEDGGN QGADGVWFGF HRGLPFLGVS 451 DGIALRHAV* ORF34ng and ORF34-1 show 90.0% identity in 459 aa overlap: 10 20 30 40 4 50 orf34-1. pep MMMPFIMLPWIAGVPAVPGQNRLSRISLWGLGGVFFGVSGLVWFSLGVS LGCAC II I II I II I II I I II II I I I : II II I II I II : II II I I II II II I I I II I II I I orf34ng MMMPFIMLPWIAGVPAVPGQKRLSRISLWGLAGVFFGVSGLVWFSLGVSFSLGVSLGCAC 10 20 30 40 50 60 60 70 80 90 100 110 orf34-1. pep FSGVSFRGSGRGTFVGSTGVSLSVFSACVPASSGCLSVXAVSAGCGLTRFFLGAAGDGSP I I I II I I II I M I II II I I I I I I I II II I: : :: Ml I I I I I II I I II II I II orf34ng FSGVSFRGSGWGAFVGSTGVSLSVFSACVPVPVNESAARAASEGRGLTRFFLGAAGDGSP 70 30 90 100 110 120 120 130 140 150 160 170 orf34-1. pep LPLSSVPSGCAGSDEAAWWCSGWAASCPTTPFGSQNSVSRGLSVCCGSAXRVLSPFGLNV I II II II I I I II I I I I I I II I I I I II II I : II I II I II II I II I I II I : I I I II I I II I orf34ng LPLSSVPSGCAGSDEAAWWCSGWAASCPTAPFGSQNSVSRGLSVCCGSVWRVLSPFGLNV 130 140 150 160 170 180 180 190 200 210 220 230 orf34-1. pep LTMPIANAPMAAIQMSNTARIRSLGVSLKGLFGFFAILIVLLGCRAMPSEGGSDGIAESA II II I I I I I I : I II I I II II I II I I II I I I II I II I I II I I II II II I I I II I I II I II orf34ng LTMPTANAPMAVIQMSNTARIRSLGVSLKGLFGFFAILIVLLGCRAMPSEGGSDGIAESA 190 200 210 220 230 240 240 250 260 270 280 290 orf34-1. pep LDWLVEGDDFLYADGGADFLGNLRLFFGGEDAHNVGYVAVGNDFDARLCGGADAQQRGA I I II II I I : I II I I I I I II II I II I I II I II II I II I I : I II II II I I I I : I II II II I I orf34ng LDWLVEGNDFLYADGGADFLGNLRLFFGGEDAHNVGYIAVGNDFDARLCSGADAQQRGA 250 260 270 280 290 300 300 310 320 330 340 350 orf34-1. pep DFGCVPSVAGDVAGSARQGGDGNIWHAFGGLFGTCNLTDELFFAFGGDLSEQQQVAWA III I I I II I I II I II I I II I I : I I: II I I I I I II I II I II I II II I I II I I I I I I I II orf34ng DFGRVPSVAGDVARSARQGGDGNVWYAFGGLFGTCNLTDELFFAFGGDLSEQQQVAWA 310 320 330 340 350 360 360 370 380 390 400 410 orf34-1. pep DDGDLGRVAFGLWLAQIGTGGGFDTQRHNVWGLRAGGSAVDGGFRADGGASDYCADAA I II II II II II II II II : I II I II I II II II I : II II II II I I II II I I : I IMM orf34ng DDGDLGRVAFGLVVLAQVGTGGGFDTQRHNWIGLRAGGSAVDDGFCADGGPADDCAEAA 370 380 390 400 410 420 420 430 440 450 orf34-1. pep AKGKAENGGNQGADGVRFGFHRVLPFLGVSDGIALRHAVX M II II : I I II I I II I I I II I I I I I II II I II I II I I I orf34ng AEGKAEDGGNQGADGVWFGFHRGLPFLGVSDGIALRHAVX 430 440 450 460 Based on this analysis, including the presence of a putative leader sequence (double-underlined) and several putative transmembrane domains (single-underlined) in the gonococcal protein, it is predicted that the proteins from N. meningitidis and N.gonorrhoeae, and their epitopes, could be useful antigens for vaccines or diagnostics, or for raising antibodies. Example 26 The following partial DNA sequence was identified in N. meningitidis <SEQ ID 215>: 1 ATGAAAACCT TCTTCAAAAC CCTTTCCGCC GCCGCACTCG CGCTCATCCT 51 CGCCGCCTGC GGATT . CAAA AAGACAGCGC GCCCGCCGCA TCCGCTTCTG 101 CCGCCGCCGA CAACGGCGCG GCGTAAAAAA GAAATCGTCT TCGGCACGAC 151 CGTCGGCGAC TTCGGCGATA TGGTCAAAGA ACAAATCCAA GCCGAGCTGG 201 AGAAAAAAGG CTACACCGTC AAACTGGTCG AGTTTACCGA CTATGTACGC 251 CCGAATCTGG CATTGGCTGA GGGCGAGTTG This corresponds to the amino acid sequence <SEQ ID 216; ORF4>: 1 MKTFFKTLSA AALALILAAC G.QKDSAPAA SASAAADNGA AKKEIVFGTT 51 VGDFGDMVKE QIQAELEKKG YTVKLVEFTD YVRPNLALAE GEL Further sequence analysis revealed the complete nucleotide sequence <SEQ ID 217>: 1 ATGAAAACCT TCTTCAAAAC CCTTTCCGCC GCCGCACTCG CGCTCATCCT 51 CGCCGCCTGC GGCGGTCAAA AAGACAGCGC GCCCGCCGCA TCCGCTTCTG 101 CCGCCGCCGA CAACGGCGCG GCGAAAAAAG AAATCGTCTT CGGCACGACC 151 GTCGGCGACT TCGGCGATAT GGTCAAAGAA CAAATCCAAG CCGAGCTGGA 201 GAAAAAAGGC TACACCGTCA AACTGGTCGA GTTTACCGAC TATGTACGCC 251 CGAATCTGGC ATTGGCTGAG GGCGAGTTGG ACATCAACGT CTTCCAACAC 301 AAACCCTATC TTGACGACTT CAAAAAAGAA CACAATCTGG ACATCACCGA 351 AGTCTTCCAA GTGCCGACCG CGCCTTTGGG ACTGTACCCG GGCAAGCTGA 401 AATCGCTGGA AGAAGTCAAA GACGGCAGCA CCGTATCCGC GCCCAACGAC 451 CCGTCCAACT TCGCCCGCGT CTTGGTGATG CTCGACGAAC TGGGTTGGAT 501 CAAACTCAAA GACGGCATCA ATCCGTTGAC CGCATCCAAA GCGGACATCG 551 CCGAGAACCT GAAAAACATC AAAATCGTCG AGCTTGAAGC CGCGCAACTG 601 CCGCGTAGCC GCGCCGACGT GGATTTTGCC GTCGTCAACG GCAACTACGC 651 CATAAGCAGC GGCATGAAGC TGACCGAAGC CCTGTTCCAA GAACCGAGCT 701 TTGCCTATGT CAACTGGTCT GCCGTCAAAA CCGCCGACAA AGACAGCCAA 751 TGGCTTAAAG ACGTAACCGA GGCCTATAAC TCCGACGCGT TCAAAGCCTA 801 CGCGCACAAA CGCTTCGAGG GCTACAAATC CCCTGCCGCA TGGAATGAAG 851 GCGCAGCCAA ATAA This corresponds to the amino acid sequence <SEQ ID 218; ORF4-l>: 1 MKTFFKTLSA AALALILAAC GGQKDSAPAA SASAAADNGA AKKEIVFGTT 51 VGDFGDMVKE QIQAELEKKG YTVKLVEFTD YVRPNLALAE GELDINVFQH 101 KPYLDDFKKE HNLDITEVFQ VPTAPLGLYP GKLKSLEEVK DGSTVSAPND 151 PSNFARVLVM LDELGWIKLK DGINPLTASK ADIAENLKNI KIVELEAAQL 201 PRSRADVDFA WNGNYAISS GMKLTEALFQ EPSFAYVNWS AVKTADKDSQ 251 WLKDVTEAYN SDAFKAYAHK RFEGYKSPAA WNEGAAK* Computer analysis of this amino acid sequence gave the following results: Homology with a predicted ORF from N.meninsitidis (strain Al ORF4 shows 93.5% identity over a 93aa overlap with an ORF (ORF4a) from strain A of N meningitidis: 10 20 30 40 50 59 orf .pep MKTFFKTLSAAALALILAACG-QKDSAPAASASAAADΝGAAKKEIVFGTTVGDFGDMVKE I I I II II I II I II I I II II I I I I II II I II II II II II II II II I I II II II II I I I I orf4a MKTFFKTLSAAALALILAACGGQKDSAPAASASAAADΝGAAXKEIVFGTTVGDFGDMVKE 10 20 30 40 50 60 60 70 80 90 orf4. pep QIQAELEKKGYTVKLVEFTDYVRPΝLALAEGEL II II I II II I II II I I I I I I I I II II II I orf4a XIQPELEKKGYTVKLVEXTDYVRXΝLALAEGELDIΝVXQHXXYLDDXKKXHΝLDITXVXQ 70 80 90 100 110 120 orf4a VPTAPLGLYPGKLKSLXXVKXGSTVSAPΝDPXXFXRVLVMLDELGXIKLKDXIXXXXXXX 130 140 150 160 170 180 The complete length ORF4a nucleotide sequence <SEQ ID 219> is: 1 ATGAAAACCT TCTTCAAAAC CCTTTCCGCC GCCGCACTCG CGCTCATCCT 51 CGCCGCCTGC GGCGGTCAAA AAGATAGCGC GCCCGCCGCA TCCGCTTCTG 101 CCGCCGCCGA CAACGGCGCG GCGAAΝAAAG AAATCGTCTT CGGCACGACC 151 GTCGGCGACT TCGGCGATAT GGTCAAAGAA CAΝATCCAAC CCGAGCTGGA 201 GAAAAAAGGC TACACCGTCA AACTGGTCGA GTΝTACCGAC TATGTGCGCΝ 251 CGAATCTGGC ATTGGCTGAG GGCGAGTTGG ACATCAACGT CTTΝCAACAC 301 AΝACΝCTATC TTGACGACTΝ CAAAAAAΝAA CACAATCTGG ACATCACCΝΝ 351 AGTCTTΝCAA GTGCCGACCG CGCCTTTGGG ACTGTACCCG GGCAAGCTGA 401 AATCGCTGGA ΝΝAAGTCAAA GAΝGGCAGCA CCGTATCCGC GCCCAACGAC 451 CCGTΝΝΝACT TCGΝCCGCGT CTTGGTGATG CTCGACGAAC TGGGTTΝGAT 501 CAAACTCAAA GACΝGCATCA ΝΝΝΝGΝΝGΝΝ ΝΝΝAΝCΝAΝA ΝΝΝGAΝAΝΝΝ 551 ΝΝΝΝAΝΝΝΝT ΝΝΝΝΝΝΝΝΝΝ ΝΝΝΝΝCΝΝCG ΝΝΝΝΝΝΝAΝΝ ΝΝΝΝΝΝΝΝΝΝ 601 ΝCGΝΝTΝΝΝΝ ΝΝGCΝΝΝΝΝT ΝΝAΝΝΝTΝΝΝ ΝΝCΝΝCΝΝΝΝ ΝΝΝΝΝTΝΝΝΝ 651 ΝAΝΝAΝΝAGC GGCATGAAGC TGACCGAAGC CCTGTTCCAA GAACCGAGCT 701 TTGCCTATGT CAACTGGTCT GCCGTCAAAA CCGCCGACAA AGACAGCCAA 751 TGGCTTAAAG ACGTAACCGA GGCCTATAAC TCCGACGCGT TCAAAGCCTA 801 CGCGCACAAA CGCTTCGAGG GCTACAAATC CCCTGCCGCA TGGAATGAAG 851 GCGCAGCCAA ATAA This is predicted to encode a protein having amino acid sequence <SEQ ID 220>: 1 MKTFFKTLSA AALALILAAC GGQKDSAPAA SASAAADNGA AXKEIVFGTT 51 VGDFGDMVKE XIQPELEKKG YTVKLVEXTD YVRXNLALAE GELDINVXQH 101 XXYLDDXKKX HNLDITXVXQ VPTAPLGLYP GKLKSLXXVK XGSTVSAPND 151 PXXFXRVLVM LDELGXIKLK DXIXXXXXXX XXXXXXXXXX XXXXXXXXXX 201 XXXXAXXXXX XXXXXXXXXS GMKLTEALFQ EPSFAYVNWS AVKTADKDSQ 251 WLKDVTEAYN SDAFKAYAHK RFEGYKSPAA WNEGAAK* A leader peptide is underlined. Further analysis of these strain A sequences revealed the complete DNA sequence <SEQ LD 221 >: 1 ATGAAAACCT TCTTCAAAAC CCTTTCCGCC GCCGCACTCG CGCTCATCCT 51 CGCCGCCTGC GGCGGTCAAA AAGATAGCGC GCCCGCCGCA TCCGCTTCTG 101 CCGCCGCCGA CAACGGCGCG GCGAAAAAAG AAATCGTCTT CGGCACGACC 151 GTCGGCGACT TCGGCGATAT GGTCAAAGAA CAAATCCAAC CCGAGCTGGA 201 GAAAAAAGGC TACACCGTCA AACTGGTCGA GTTTACCGAC TATGTGCGCC 251 CGAATCTGGC ATTGGCTGAG GGCGAGTTGG ACATCAACGT CTTCCAACAC 301 AAACCCTATC TTGACGACTT CAAAAAAGAA CACAATCTGG ACATCACCGA 351 AGTCTTCCAA GTGCCGACCG CGCCTTTGGG ACTGTACCCG GGCAAGCTGA 401 AATCGCTGGA AGAAGTCAAA GACGGCAGCA CCGTATCCGC GCCCAACGAC 451 CCGTCCAACT TCGCCCGCGT CTTGGTGATG CTCGACGAAC TGGGTTGGAT 501 CAAACTCAAA GACGGCATCA ATCCGCTGAC CGCATCCAAA GCGGACATTG 551 CCGAAAACCT GAAAAACATC AAAATCGTCG AGCTTGAAGC CGCGCAACTG 601 CCGCGTAGCC GCGCCGACGT GGATTTTGCC GTCGTCAACG GCAACTACGC 651 CATAAGCAGC GGCATGAAGC TGACCGAAGC CCTGTTCCAA GAACCGAGCT 701 TTGCCTATGT CAACTGGTCT GCCGTCAAAA CCGCCGACAA AGACAGCCAA 751 TGGCTTAAAG ACGTAACCGA GGCCTATAAC TCCGACGCGT TCAAAGCCTA 801 CGCGCACAAA CGCTTCGAGG GCTACAAATC CCCTGCCGCA TGGAATGAAG 851 GCGCAGCCAA ATAA This encodes a protein having amino acid sequence <SEQ ID 222; ORF4a-l>: 1 MKTFFKTLSA AALALILAAC GGQKDSAPAA SASAAADNGA AKKEIVFGTT 51 VGDFGDMVKE QIQPELEKKG YTVKLVEFTD YVRPNLALAE GELDINVFQH 101 KPYLDDFKKE HNLDITEVFQ VPTAPLGLYP GKLKSLEEVK DGSTVSAPND 151 PSNFARVLVM LDELGWIKLK DGINPLTASK ADIAENLKNI KIVELEAAQL 201 PRSRADVDFA WNGNYAISS GMKLTEALFQ EPSFAYVNWS AVKTADKDSQ 251 WLKDVTEAYN SDAFKAYAHK RFEGYKSPAA WNEGAAK* ORF4a-l and ORF4-1 show 99.7% identity in 287 aa overlap: 10 20 30 40 50 60 orf4a-l MKTFFKTLSAAALALILAACGGQKDSAPAASASAAADNGAAKKEIVFGTTVGDFGDMVKE I II I II II II I II II I II I I II I II I I II II I II II II I I I I I I I I II I I I I I I I I I II I orf4-1 MKTFFKTLSAAALALILAACGGQKDSAPAASASAAADNGAAKKEIVFGTTVGDFGDMVKE 10 20 30 40 50 60 70 80 90 100 110 120 orf4a-l QIQPELEKKGYTVKLVEFTDYVRPNLALAEGELDINVFQHKPYLDDFKKEHNLDITEVFQ III II II II I II II I II I I I II I I II II I II I II II II II II II I I I I I II I II II I I I orf4-1 QIQAELEKKGYTVKLVEFTDYVRPNLALAEGELDINVFQHKPYLDDFKKEHNLDITEVFQ 70 80 90 100 110 120 130 140 150 160 170 180 orf4a-l VPTAPLGLYPGKLKSLEEVKDGSTVSAPNDPSNFARVLVMLDELGWIKLKDGINPLTASK II I II II I I I I I I II I I I I I I II I I II II I II I II I I II II II II II I II II I II II I I I orf4-1 VPTAPLGLYPGKLKSLEEVKDGSTVSAPNDPSNFARVLVMLDELGWIKLKDGINPLTASK 130 140 150 160 170 180 190 200 210 220 230 240 orf4a-l ADIAENLKNIKIVELEAAQLPRSRADVDFAWNGNYAISSGMKLTEALFQEPSFAYVNWS I II I II I I I II I II II I I II II I II I II I II I II I II II I I II I I I I I I I II I II II I I I orf4-1 ADIAENLKNIKIVELEAAQLPRSRADVDFAWNGNYAISSGMKLTEALFQEPSFAYVNWS 190 200 210 220 230 240 250 260 270 280 orf4a-l AVKTADKDSQWLKDVTEAYNSDAFKAYAHKRFEGYKSPAAWNEGAAKX II I II II I II I II I M I M I I I I II I II I II I II I I II I I M M M I I orf4-1 AVKTADKDSQWLKDVTEAYNSDAFKAYAHKRFEGYKSPAAWNEGAAKX 250 260 270 280 Homology with an outer membrane protein of Pasteur ella haemolitica (accession q08869). ORF4 and this outer membrane protein show 33% aa identity in 91aa overlap: 10 20 lip2.pasha MNFKKLLGVALVSALALTACKDEKAQAP II I :: I I M M : |: I ORF4 VXTPNPDGRTPCPSFLFETATTSGENMKTFFKTLSAAAL—ALILAACGFKKTARPPHPL 110 120 130 140 150 30 40 50 60 70 80 lip2. pasha -ATTAKTENKAPLKVGVMTGPEAQMTEVAVKIAKEKYGLDVELVQFTEYTQPNAALHSKD : :: I : :: h: :: II I I : I I : I I : I : : II II : ORF4 LPPPTTARRKKEIVFGTTVGDFGDMVKEQIQAELEKKGYTVKLVEFTDYVRPNLALAEGE 160 170 180 190 200 210 90 100 110 120 130 140 lip2. pasha LDANAFQTVPYLEQEVKDRGYKLAIIGNTLVWPIAAYSKKIKNISELKDGATVAIPNNAS I ORF4 L Homology with a predicted ORF from N.sonorrhoeae ORF4 shows 93.6% identity over a 94aa overlap with a predicted ORF (ORF4.ng) from N gonorrhoeae: 10 20 30 orf nm.pep MKTFFKTLSAAALALILAACGXQKDSAPAA II II I I I IM I: I II I I II I I I I I I I I I I orf4ng RAΝAVXTPΝPDGRTPCLSFLFETATTSGEΝMKTFFKTLSTASLALILAACGGQKDSAPAA 200 210 220 230 240 250 40 50 60 70 80 89 orf4nm.pep SASA-AADΝGAAKKEIVFGTTVGDFGDMVKEQIQAELEKKGYTVKLVEFTDYVRPΝLALA IM I : II II II I I II II II II I II I II II II II I II II II II II I II I I II II I I I I I I orf4ng SAAAPSADΝGAAKKEIVFGTTVGDFGDMVKEQIQAELEKKGYTVKLVEFTDYVRPΝLALA 260 270 280 290 300 310 90 orf4nm.pep EGEL I I II orf4ng EGELDIΝVFQHKPYLDDFKKEHΝLDITEAFQVPTAPLGLYPGKLKSLEEVKDGSTVSAPΝ 320 330 340 350 360 370 The complete length ORF4ng nucleotide sequence <SEQ ID 223> was predicted to encode a protein having amino acid sequence <SEQ ID 224>: 1 MKTFFKTLST ASLALIL AC GGQKDSAPAA SAAAPSADΝG AAKKEIVFGT 51 TVGDFGDMVK EQIQAELEKK GYTVKLVEFT DYVRPΝLALA EGELDIΝVFQ 101 HKPYLDDFKK EHΝLDITEAF QVPTAPLGLY PGKLKSLEEV KDGSTVSAPΝ 151 DPSΝFARALV MLΝELGWIKL KDGIΝPLTAS KADIAEΝLKΝ IKIVELEAAQ 201 LPRSRADVDF AWΝGΝYAIS SGMKLTEALF QEPSFAYVΝW SAVKTADKDS 251 QWLKDVTEAY ΝSDAFKAYAH KRFEGYKYPA AWΝEGAAK* Further analysis revealed the complete length ORF4ng DΝA sequence <SEQ ID 225> to be: 1 atgAAAACCT TCTTCAAAAC cctttccgcc gccgcaCTCG CGCTCATCCT 51 CGCAGCCTGc ggCggtcaAA AAGACAGCGC GCCCgcagcc tctgcCGCCG 101 CCCCTTCTGC CGATAACGgc gCgGCGAAAA AAGAAAtcgt ctTCGGCACG 151 Accgtgggcg acttcggcgA TAtggTCAAA GAACAAATCC AagcCGAgct 201 gGAGAAAAAA GgctACACcg tcAAattggt cgaatttacc gactatgtGC 251 gCCCGAATCT GGCATTGGCG GAGGGCGAGT TGGACATCAA CGTCTTCCAA 301 CACAAACCCT ATCTTGACGA TTTCAAAAAA GAACACAACC TGGACATCAC 351 CGAAGCCTTC CAAGTGCCGA CCGCGCCTTT GGGACTGTAT CCGGGCAAAC 401 TGAAATCGCT GGAAGAAGTC AAAGACGGCA GCACCGTATC CGCGCCCAac 451 gACccgTCCA ACTTCGCACG CGCCTTGGTG ATGCTGAACG AACTGGGTTG 501 GATCAAACTC AAAGACGGCA TCAATCCGCT GACCGCATCC AAAGCCGACA 551 TCGCGGAAAA CCTGAAAAAC ATCAAAATCG TCGAGCTTGA AGCCGCACAA 601 CTGCCGCGCA GCCGCGCCGA CGTGGATTTT GCCGTCGTCA ACGGCAACTA 651 CGCCATAAGC AGCGGCATGA AGCTGACCGA AGCCCTGTTC CAAGAGCCGA 701 GCTTTGCCTA TGTCAACTGG TCTGCCgtcA AAACCGCCGA CAAAGACAGC 751 CAATGGCTTA AAGACGTAAC CGAGGCCTAT AACTCCGACG CGTTCAAAGC 801 CTACGCGCAC AAACGCTTCG AGGGCTACAA ATACCCTGCC GCATGGAATG 851 AAGGCGCAGC CAAATAA This encodes a protein having amino acid sequence <SEQ ID 226; ORF4ng-l>: 1 MKTFFKTLSA AALALILAAC GGQKDSAPAA SAAAPSADNG AAKKEIVFGT 51 TVGDFGDMVK EQIQAELEKK GYTVKLVEFT DYVRPNLALA EGELDINVFQ 101 HKPYLDDFKK EHNLDITEAF QVPTAPLGLY PGKLKSLEEV KDGSTVSAPN 151 DPSNFARALV MLNELGWIKL KDGINPLTAS KADIAENLKN IKIVELEAAQ 201 LPRSRADVDF AWNGNYAIS SGMKLTEALF QEPSFAYVNW SAVKTADKDS 251 QWLKDVTEAY NSDAFKAYAH KRFEGYKYPA AWNEGAAK* This shows 97.6% identity in 288 aa overlap with ORF4-1: 10 20 30 40 50 59 orf4-1. pep MKTFFKTLSAAALALILAACGGQKDSAPAASASA-AADNGAAKKEIVFGTTVGDFGDMVK II I II II I II I II I I I I I II I I II I II II II I : I M I II I II II I I I I II II I I I II I I orf4ng-l MKTFFKTLSAAALALILAACGGQKDSAPAASAAAPSADNGAAKKEIVFGTTVGDFGDMVK 10 20 30 40 50 60 60 70 80 90 100 110 119 orf4-1. pep EQIQAELEKKGYTVKLVEFTDYVRPNLALAEGELDINVFQHKPYLDDFKKEHNLDITEVF I I II I II I II II I I I I II II I I II I II II I II I II II II I II II I I I I II I I II I I I I : I orf4ng-l EQIQAELEKKGYTVKLVEFTDYVRPNLALAEGELDINVFQHKPYLDDFKKEHNLDITEAF 70 80 90 100 110 120 120 130 140 150 160 170 179 orf4-1. pep QVPTAPLGLYPGKLKSLEEVKDGSTVSAPNDPSNFARVLVMLDELGWIKLKDGINPLTAS II I II I II I I I II I II II I I II I II I II II I II I I I I : I I II : I II I I I I II I I II II I I orf4ng-l QVPTAPLGLYPGKLKSLEEVKDGSTVSAPNDPSNFARALVMLNELGWIKLKDGINPLTAS 130 140 150 160 170 180 180 190 200 210 220 230 239 orf4-1. pep KADIAENLKNIKIVELEAAQLPRSRADVDFAWNGNYAISSGMKLTEALFQEPSFAYVNW I I I II I I I I I I II I II II I II II I II I I I II II I I I I I I I I I II II I I I II II I I I II I I orf4ng-l KADIAENLKNIKIVELEAAQLPRSRADVDFAWNGNYAISSGMKLTEALFQEPSFAYVNW 190 200 210 220 230 240 240 250 260 270 280 orf4-1. pep SAVKTADKDSQWLKDVTEAYNSDAFKAYAHKRFEGYKSPAAWNEGAAKX I II I I I I I II II I I I I II II I I I II I II II II I I II I I II I II II I I I orf4ng-l SAVKTADKDSQWLKDVTEAYNSDAFKAYAHKRFEGYKYPAAWNEGAAKX 250 260 270 280 In addition, ORF4ng-l shows significant homology with an outer membrane protein from the database: ID LLIIPP22 PPAASSHHAA SSTTAANNDDAARRDD;; PRT; 276 AA. AC Q08869; DT 01-NOV-1995 (REL. 3322,, CCRREATED) DT 01-NOV-1995 (REL. 3322,, LLAAST SEQUENCE UPDATE) DT 01-NOV-1995 (REL. 3322,, LLAAST ANNOTATION UPDATE) DE 28.2 KD OUTER MEMBRANE PROTEIN PRECURSOR. . . . SCORES Initl: 279 Initn: 416 Opt: 494 Smith-Waterman score: 494; 36.0% identity in 275 aa overlap 10 20 30 40 50 orf4ng-l . pep MKTFFKTLSAAAL—ALILAACGGQKDSAPAASAAAPSADNGAAKKEIVFGTTVGDFGDM I I I :: I I M MM : I : I II :: I :::| I I M M :: I lip2_pasha MNFKKLLGVALVSALALTACKDEKAQAPATTA KTENKAPLK VGVMTGPEAQM 10 20 30 40 50 60 70 80 90 100 110 orf ng-l . pep VKEQIQAELEKKGYTVKLVEFTDYVRPNLALAEGELDINVFQHKPYLDDFKKEHNLDITE : : :: II I I : II : I I : I : : I I II : I I I : I I III:: I ::: :: lip2_pasha TEVAVKIAKEKYGLDVELVQFTEYTQPNAALHSKDLDANAFQTVPYLEQEVKDRGYKLAI 60 70 80 90 100 110 120 130 140 150 160 170 orf4ng-l .pep AFQVPTAPLGLYPGKLKSLEEVKDGSTVSAPNDPSNFARALVMLNELGWIKLKDGINPLT :: : I :: I I : I : : I : II I : II : II : II I II I : : I : I : II II I : lip2_pasha IGNTLVWPIAAYSKKIKNISELKDGATVAIPNNASNTARALLLLQAHGLLKLKDPKN-VF 120 130 140 150 160 170 180 190 200 210 220 230 orf4ng-l . ep ASKADIAENLKNIKIVELEAAQLPRSRADVDFAVVNGNYAISSGMKLTE—ALFQEPSFA lip2_pasha ATENDIIENPKNIKIVQADTSLLTRMLDDVELAVINNTYAGQAGLSPDKDGIIVESKDSP 180 190 200 210 220 230 240 250 260 270 280 289 orf4ng-l . pep YVNWSAVKTADKDSQWLKDVTEAYNSDAFKAYAHKRFEGYKYPAAWNEGAAKX III : : M M ::::::: I I M I lip2_pasha YVNLWSREDNKDDPRLQTFVKSFQTEEVFQEALKLFNGGVVKGW 240 250 260 270 Based on this analysis, including the homology with the outer membrane protein of Pasteurella haemolitica, and on the presence of a putative prokaryotic membrane lipoprotein lipid attachment site in the gonococcal protein, it was predicted that these proteins from N. meningitidis .and N.gonorrhoeae, and their epitopes, could be useful antigens for vaccines or diagnostics, or for raising antibodies. ORF4-1 (30kDa) was cloned in pET and pGex vectors and expressed in E.coli, as described above. The products of protein expression and purification were analyzed by SDS-PAGE. Figures 8 A and 8B show, repsectively, the results of affinity purification of the His-fusion and GST-fusion proteins. Purified His-fusion protein was used to immunise mice, whose sera were used for ELISA (positive result), Western blot (Figure 8C), FACS analysis (Figure 8D), and a bactericidal assay (Figure 8E). These experiments confirm that ORF4-1 is a surface-exposed protein, and that it is a useful immunogen. Figure 8F shows plots of hydrophilicity, antigenic index, and AMPHI regions for ORF4-1. Example 27 The following partial DNA sequence was identified in N. meningitidis <SEQ ID 227>: 1 CCTCGTCGTC CTCGGCATGC TCCAGTTTCA AGGGGCGATT TACTCCAAGG 51 CGGTGGAACG TATGCTCGGC ACGGTCATCG GGCTGGGCGC GGGTTTGGGC 101 GTTTTATGGC TGAACCAGCA TTATTTCCAC GGCAACCTCC TCTTCTACCT 151 CACCGTCGGC ACGGCAAGCG CACTGGCCGG CTGGGCGGCG GTCGGCAAAA 201 ACGGCTACGT CCCTmTGCTG GCAGGGCTGA CGATGTGTAT GCTCATCGGC 251 GACAACGGCA GCGAATGGCT CGACAGCGGA CTCATGCGCG CCATGAACGT 301 CCTCATCGGC GyGGCCATCG CCATCGCCGC CGCCAAACTG CTGCCGCTGA 351 AATCCACACT GATGTGGCGT TTCATGCTTG CCGACAACCT GGCCGACTGC 401 AGCAAAATGA TTGCCGAAAT CAGCAACGGC AGGCGCATGA CCCGCGAACG 451 CCTCGAGGAG AACATGGCGA AAATGCGCCA AATCAACGCA CGCATGGTCA 501 AAAGCCGCAG CCATCTCGCC GCCACATCGG GCGAAAGCTG CATCAGCCCC 551 GCCATGATGG AAGCCATGCA GCACGCCCAC CGTAAAATCG TCAACACCAC 601 CGAGCTGCTC CTGACCACCG CCGCCAAGCT GCAATCTCCC AAACTCAACG 651 GCAGCGAAAT CCGGCTGCTT GACCGCCACT TCACACTGCT CCAAAC ... 701 GC AGACACGCCC GCCGCATCCG 751 CATCGACACC GCCATCAACC CCGAACTGGA AGCCCTCGCC GAACACCTCC 801 ACTACCAATG GCAGGGCTTC CTCTGGCTCA GCACCGATAT GCGTCAGGAA 851 ATTTCCGCCC TCGTCATCCT GCTGCAACGC ACCCGCCGCA AATGGCTGGA 901 TGCCCACGAA CGCCAACACC TGCGCCAAAG CCTGCTTGA This corresponds to the .amino acid sequence <SEQ ID 228; ORF8>: 1 PRRP RHAPVSRGDL LQGGGTYARH GHRAGRGFGR FMAEPALFPR 51 QPPLLPHRRH GKRTGRLGGG RQKRLRPXAG RADDVYAHRR QRQRMARQRT 101 HARHERPHRR GHRHRRRQTA AAEIHTDVAF HACRQPGRLQ QNDCRNQQRQ 151 AHDPRTPRGE HGENAPNQRT HGQKPQPSRR HIGRKLHQPR HDGSHAARPP 201 XNRQHHRAAP DHRRQAAISQ TQRQRNPAAX PPLHTAPN Q 251 TRPPHPHRHR HQPRTGSPRR TPPLPMAGLP LAQHRYASGN FRPRHPAATH 301 PPQMAGCPRT PTPAPKPA* Computer analysis of this amino acid sequence gave the following results: Sequence motifs ORF8 is proline-rich and has a distribution of proline residues consistent with a surface localization. Furthermore the presence of an RGD motif may indicate a possible role in bacterial adhesion events. Homology with a predicted ORF from N.gonorrhoeae ORF8 shows 86.5% identity over a 312aa overlap with a predicted ORF (ORF8.ng) from N gonorrhoeae: orfθng 1 MDRDDRLRRPRHAPVPRRDLLQRGGTYARYGHRAGRGFGRFMAEPALFPR 50 II I II II I I I I II II I I II : II I I II I I II I I I II I I II I orf8. pep 1 PRRPRHAPVSRGDLLQGGGTYARHGHRAGRGFGRFMAEPALFPR 44 orf8ng 51 QPPLLPDHRHGKRTGRLGGGRQKRLRPYVGGADDVHAHRRQRQRMARQRP 100 II I II I II I II II I I I II I II I I I I I II II : I II II I II II I I I orf8. pep 45 QPPLLPHRRHGKRTGRLGGGRQKRLRPXAGRADDVYAHRRQRQRMARQRT 94 orfδng 101 DARDERPHRRRHRHCRRQTAAAEIHTDVAFHACRQPGRLQQΝDCRΝQQRQ 150 II I I II I I III I II I I II I II II I I I II I I II I I II II II I II II orf8. pep 95 HARHERPHRRGHRHRRRQTAAAEIHTDVAFHACRQPGRMQQΝDCRΝQQRQ 144 orfδng 151 AYDARTFGAEYGQΝAPΝQRTHGQKPQPPRRHIGRKPHQPLHDGSHAARPP 200 I : I II I : I : II II I II I II I II I II I II II III I I I I II I I I I orf8. pep 145 AHDPRTPRGEHGEΝAPΝQRTHGQKPQPSRRHIGRKLHQPRHDGSHAARPP 194 orf8ng 201 QΝRQHHRAAPDHRRQAAISQTQRQRΝPAARPPLHTAPΝRPATΝRRPHQRQ 250 II II I II I I I II I II II I I II I II I I I I II II II II I orf8. pep 195 XΝRQHHRAAPDHRRQAAISQTQRQRΝPAAXPPLHTAPΝ Q 244 orfδng 251 TRPPHPHRHRHQPRTGSPRRTPPLPMAGFPLAQHQYASGΝFRPRHPPATH 300 I II II I II I II I II I II II I I II I II II I II I I . II I I I II I I I I III orf8. pep 245 TRPPHPHRHRHQPRTGSPRRTPPLPMAGLPLAQHRYASGΝFRPRHPAATH 294 orfδng 301 PPQMAGCPRTPTPAPKPA* 319 I I I I II I II I II I II I II I orfθ.pep 295 PPQMAGCPRTPTPAPKPA* 313 The complete length ORF8ng nucleotide sequence <SEQ ID 229> is predicted to encode a protein having amino acid sequence <SEQ ID 230>: 1 MDRDDRLRRP RHAPVPRRDL LQRGGTYARY GHRAGRGFGR FMAEPALFPR 51 QPPLLPDHRH GKRTGRLGGG RQKRLRPYVG GADDVHAHRR QRQRMARQRP 101 DARDERPHRR RHRHCRRQTA AAEIHTDVAF HACRQPGRLQ QΝDCRΝQQRQ 151 AYDARTFGAE YGQΝAPΝQRT HGQKPQPPRR HIGRKPHQPL HDGSHAARPP 201 QNRQHHRAAP DHRRQAAISQ TQRQRNPAAR PPLHTAPNRP ATNRRPHQRQ 251 TRPPHPHRHR HQPRTGSPRR TPPLPMAGFP LAQHQYASGN FRPRHPPATH 301 PPQMAGCPRT PTPAPKPA* Based on the sequence motifs in these proteins, it is predicted that the proteins from N. meningitidis and N.gonorrhoeae, and their epitopes, could be useful antigens for vaccines or diagnostics, or for raising antibodies. Example 28 The following partial DNA sequence was identified in N. meningitidis <SEQ ID 231>: 1 .. GAAATCAGCC TGCGGTCCGA CNACAGGCCG GTTTCCGTGN CGAAGCGGCG 51 GGATTCGGAA CGTTTTCTGC TGTTGGACGG CGGCAACAGC CGGCTCAAGT 101 GGGCGTGGGT GGAAAACGGC ACGTTCGCAA CCGTCGGTAG CGCGCCGTAC 151 CGCGATTTGT CGCCTTTGGG CGCGGAGTGG GCGGAAAAGG CGGATGGAAA 201 TGTCCGCATC GTCGGTTGCG CTGTGTGCGG AGAATTCAAA AAGGCACAAG 251 TGCAGGAACA GCTCGCCCGA AAAATCGAGT GGCTGCCGTC TTCCGCACAG 301 GCTTT.GGCA TACGCAACCA CTACCGCCAC CCCGAAGAAC ACGGTTCCGA 351 CCGCTGGTTC AACGCCTTGG GCAGCCGCCG CTTCAGCCGC AACGCCTGCG 401 TCGTCGTCAG TTGCGGCACG GCGGTAACGG TTGACGCGCT CACCGATGAC 451 GGACATTATC TCGGAGA.GG AACCATCATG CCCGGTTTCC ACCTGATGAA 501 AGAATCGCTC GCCGTCCGAA CCGCCAACCT CAACCGGCAC GCCGGTAAGC 551 GTTATCCTTT CCCGACCGG.. This corresponds to the amino acid sequence <SEQ ID 232; ORF61>: 1 ..EISLRSDXRP VSVXKRRDSE RFLLLDGGNS RLKWAWVENG TFATVGSAPY 51 RDLSPLGAEW AEKADGNVRI VGCAVCGEFK KAQVQEQLAR KIEWLPSSAQ 101 AXGIRNHYRH PEEHGSDRWF NALGSRRFSR NACVWSCGT AVTVDALTDD 151 GHYLGXGTIM PGFHLMKESL AVRTANLNRH AGKRYPFPT.. Further work revealed the complete nucleotide sequence <SEQ ID 233>: 1 ATGACGGTTT TGAAGCTTTC GCACTGGCGG GTGTTGGCGG AGCTTGCCGA 51 CGGTTTGCCG CAACACGTCT CGCAACTGGC GCGTATGGCG GATATGAAGC 101 CGCAGCAGCT CAACGGTTTT TGGCAGCAGA TGCCGGCGCA CATACGCGGG 151 CTGTTGCGCC AACACGACGG CTATTGGCGG CTGGTGCGCC CATTGGCGGT 201 TTTCGATGCC GAAGGTTTGC GCGAGCTGGG GGAAAGGTCG GGTTTTCAGA 251 CGGCATTGAA GCACGAGTGC GCGTCCAGCA ACGACGAGAT ACTGGAATTG 301 GCGCGGATTG CGCCGGACAA GGCGCACAAA ACCATATGCG TGACCCACCT 351 GCAAAGTAAG GGCAGGGGGC GGCAGGGGCG GAAGTGGTCG CACCGTTTGG 401 GCGAGTGTCT GATGTTCAGT TTTGGCTGGG TGTTTGACCG GCCGCAGTAT 451 GAGTTGGGTT CGCTGTCGCC TGTTGCGGCA GTGGCGTGTC GGCGCGCCTT 501 GTCGCGTTTA GGTTTGGATG TGCAGATTAA GTGGCCCAAT GATTTGGTTG 551 TCGGACGCGA CAAATTGGGC GGCATTCTGA TTGAAACGGT CAGGACGGGC 601 GGCAAAACGG TTGCCGTGGT CGGTATCGGC ATCAATTTTG TCCTGCCCAA 651 GGAAGTAGAA AATGCCGCTT CCGTGCAATC GCTGTTTCAG ACGGCATCGC 701 GGCGGGGCAA TGCCGATGCC GCCGTGCTGC TGGAAACGCT GTTGGTGGAA 751 CTGGACGCGG TGTTGTTGCA ATATGCGCGG GACGGATTTG CGCCTTTTGT 801 GGCGGAATAT CAGGCTGCCA ACCGCGACCA CGGCAAGGCG GTATTGCTGT 851 TGCGCGACGG CGAAACCGTG TTCGAAGGCA CGGTTAAAGG CGTGGACGGA 901 CAAGGCGTTT TGCACTTGGA AACGGCAGAG GGCAAACAGA CGGTCGTCAG 951 CGGCGAAATC AGCCTGCGGT CCGACGACAG GCCGGTTTCC GTGCCGAAGC 1001 GGCGGGATTC GGAACGTTTT CTGCTGTTGG ACGGCGGCAA CAGCCGGCTC 1051 AAGTGGGCGT GGGTGGAAAA CGGCACGTTC GCAACCGTCG GTAGCGCGCC 1101 GTACCGCGAT TTGTCGCCTT TGGGCGCGGA GTGGGCGGAA AAGGCGGATG 1151 GAAATGTCCG CATCGTCGGT TGCGCTGTGT GCGGAGAATT CAAAAAGGCA 1201 CAAGTGCAGG AACAGCTCGC CCGAAAAATC GAGTGGCTGC CGTCTTCCGC 1251 ACAGGCTTTG GGCATACGCA ACCACTACCG CCACCCCGAA GAACACGGTT 1301 CCGACCGCTG GTTCAACGCC TTGGGCAGCC GCCGCTTCAG CCGCAACGCC 1351 TGCGTCGTCG TCAGTTGCGG CACGGCGGTA ACGGTTGACG CGCTCACCGA 1401 TGACGGACAT TATCTCGGGG GAACCATCAT GCCCGGTTTC CACCTGATGA 1451 AAGAATCGCT CGCCGTCCGA ACCGCCAACC TCAACCGGCA CGCCGGTAAG 1501 CGTTATCCTT TCCCGACCAC AACGGGCAAT GCCGTCGCCA GCGGCATGAT 1551 GGATGCGGTT TGCGGCTCGG TTATGATGAT GCACGGGCGT TTGAAAGAAA 1601 AAACCGGGGC GGGCAAGCCT GTCGATGTCA TCATTACCGG CGGCGGCGCG 1651 GCAAAAGTTG CCGAAGCCCT GCCGCCTGCA TTTTTGGCGG AAAATACCGT 1701 GCGCGTGGCG GACAACCTCG TCATTTACGG GTTGTTGAAC ATGATTGCCG 1751 CCGAAGGCAG GGAATATGAA CATATTTAA This corresponds to the amino acid sequence <SEQ ID 234; ORF61-l>: 1 MTVLKLSHWR VLAELADGLP QHVSQLARMA DMKPQQLNGF WQQMPAHIRG 51 LLRQHDGYWR LVRPLAVFDA EGLRELGERS GFQTALKHEC ASSNDEILEL 101 ARIAPDKAHK TICVTHLQSK GRGRQGRKWS HRLGECLMFS FGWVFDRPQY 151 ELGSLSPVAA VACRRALSRL GLDVQIKWPN DLWGRDKLG GILIETVRTG 201 GKTVAWGIG INFVLPKEVE NAASVQSLFQ TASRRGNADA AVLLETLLVE 251 LDAVLLQYAR DGFAPFVAEY QAANRDHGKA VLLLRDGETV FEGTVKGVDG 301 QGVLHLETAE GKQTWSGEI SLRSDDRPVS VPKRRDSERF LLLDGGNSRL 351 KWAWVENGTF ATVGSAPYRD LSPLGAEWAE KADGNVRIVG CAVCGEFKKA 401 QVQEQLARKI EWLPSSAQAL GIRNHYRHPE EHGSDRWFNA LGSRRFSRNA 451 CWVSCGTAV TVDALTDDGH YLGGTIMPGF HLMKESLAVR TANLNRHAGK 501 RYPFPTTTGN AVASGMMDAV CGSVMMMHGR LKEKTGAGKP VDVIITGGGA 551 AKVAEALPPA FLAENTVRVA DNLVIYGLLN MIAAEGREYE HI* Figure 9 shows plots of hydrophilicity, antigenic index, and AMPHI regions for ORF61-1. Further computer analysis of this amino acid sequence gave the following results: Homology with the baf protein of B. pertussis (accession number U120201. ORF61 and baf protein show 33% aa identity in 166aa overlap: orfδl 23 LLLDGGNSRLKWAWVE-NGTFATVGSAPYR DLSPLGAEWAEKADGNVRIVGCAVCG 77 +L+D GNSRLK W + + A AP DL LG A R +G V G baf 3 ILIDSGNSRLKVGWFDPDAPQAAREPAPVAFDNLDLDALGRWLATLPRRPQRALGVNVAG 62 oorrffδδll 7 788 EFKKAQVQEQLAR KIEWLPSSAQAXGIRNHYRHPEEHGSDRW FNALGSRRFSRN 131 + + L I WL + A G+RN YR+P++ G+DRW L + baf 63 LARGEAIAATLRAGGCDIRWLRAQPLAMGLRNGYRNPDQLGADRWACMVGVLARQPSVHP 122 orfδl 1 1332 ACVWSCGTAVTVDALTDDGHYLGXGTIMPGFHLMKESLAVRTANL 177 +V S GTA T+D + D + G G I+PG +M+ +LA TA+L baf 1 1223 PLLVASFGTATTLDTIGPDNVFPG-GLILPGPAMMRGALAYGTAHL 167 Homology with a predicted ORF from N.meninsitidis (strain A) ORF61 shows 97.4% identity over a 189aa overlap with an ORF (ORFόla) from strain A of N. meningitidis: 10 20 30 orf61.pep EISLRSDXRPVSVXKRRDSERF LLDGGΝS II I II II I I I II I II I II II II II II II orf61a TVFEGTVKGVDGQGVLHLETAEGKQTWSGEISLRSDDRPVSVPKRRDSERFLLLDGGΝS 290 300 310 320 330 340 40 50 60 70 80 90 orf61. pep RLKWAWVEΝGTFATVGSAPYRDLSPLGAEWAEKADGNVRIVGCAVCGEFKKAQVQEQLAR I I I II I I I II I II II II II I I I I II II I I I I I I: I I I II II I II I I II II II II II I I I I orf61a RLKWAWVENGTFATVGSAPYRDLSPLGAEWAEKVDGNVRIVGCAVCGEFKKAQVQEQLAR 350 360 370 380 390 400 100 110 120 130 140 150 orf61.pep KIEWLPSSAQAXGIRNHYRHPEEHGSDRWFNALGSRRFSRNACVWSCGTAVTVDALTDD I II I II I II II I I II II II I II I II I I II I II I II II II I II I II II I I I II II I I II I orf61a KIEWLPSSAQALGIRNHYRHPEEHGSDRWFNALGSRRFSRNACVWSCGTAVTVDALTDD 410 420 430 440 450 460 160 170 180 189 orf61. pep GHYLGXGTIMPGFHLMKESLAVRTANLNRHAGKRYPFPT I I I I I I II II I I I II I I I II I I I I II II II I II II II I orf61a GHYLG-GTIMPGFHLMKESLAVRTANLNRHAGKRYPFPTTTGNAVASGMMDAVCGSVMMM 470 480 490 500 510 520 orf61a HGRLKEKTGAGKPVDVIITGGGAAKVAEALPPAFLAENTVRVADNLVIHGLLNLIAAEGG 530 540 550 560 570 580 The complete length ORFόla nucleotide sequence <SEQ ID 235> is: 1 ATGACGGTTT TGAAGCCTTC GCACTGGCGG GTGTTGGCGG AGCTTGCCGA 51 CGGTTTGCCG CAACACGTCT CGCAACTGGC GCGTATGGCG GATATGAAGC 101 CGCAGCAGCT CAACGGTTTT TGGCAGCAGA TGCCGGCGCA CATACGCGGG 151 CTGTTGCGCC AACACGACGG CTATTGGCGG CTGGTGCGCC CATTGGCGGT 201 TTTCGATGCC GAAGGTTTGC GCGAGCTGGG GGAAAGGTCG GGTTTTCAGA 251 CGGCATTGAA GCACGAGTGC GCGTCCAGCA ACGACGAGAT ACTGGAATTG 301 GCGCGGATTG CGCCGGACAA GGCGCACAAA ACCATATGTG TGACCCACCT 351 GCAAAGTAAG GGCAGGGGGC GGCAGGGGCG GAAGTGGTCG CACCGTTTGG 401 GCGAGTGTCT GATGTTCAGT TTTGGCTGGG TGTTTGACCG GCCGCAGTAT 451 GAGTTGGGTT CGCTGTCGCC TGTTGCGGCA GTGGCGTGCC GGCGCGCCTT 501 GTCGCGTTTG GGTTTGAAAA CGCAAATCAA GTGGCCAAAC GATTTGGTCG 551 TCGGACGCGA CAAATTGGGC GGCATTCTGA TTGAAACGGT CAGGACGGGC 601 GGCAAAACGG TTGCCGTGGT CGGTATCGGC ATCAATTTCG TGCTGCCCAA 651 GGAAGTGGAA AACGCCGCTT CCGTGCAATC GCTGTTTCAG ACGGCATCGC 701 GGCGGGGAAA TGCCGATGCC GCCGTGTTGC TGGAAACGCT GTTGGCGGAA 751 CTTGATGCGG TGTTGTTGCA ATATGCGCGG GACGGATTTG CGCCTTTTGT 801 GGCGGAATAT CAGGCTGCCA ACCGCGACCA CGGCAAGGCG GTATTGCTGT 851 TGCGCGACGG CGAAACCGTG TTCGAAGGCA CGGTTAAAGG CGTGGACGGA 901 CAAGGCGTTC TGCACTTGGA AACGGCAGAG GGCAAACAGA CGGTCGTCAG 951 CGGCGAAATC AGCCTGCGGT CCGACGACAG GCCGGTTTCC GTGCCGAAGC 1001 GGCGGGATTC GGAACGTTTT CTGCTGTTGG ACGGCGGCAA CAGCCGGCTC 1051 AAGTGGGCGT GGGTGGAAAA CGGCACGTTC GCAACCGTCG GTAGCGCGCC 1101 GTACCGCGAT TTGTCGCCTT TGGGCGCGGA GTGGGCGGAA AAGGTGGATG 1151 GAAATGTCCG CATCGTCGGT TGCGCCGTGT GCGGAGAATT CAAAAAGGCA 1201 CAAGTGCAGG AACAGCTCGC CCGAAAAATC GAGTGGCTGC CGTCTTCCGC 1251 ACAGGCTTTG GGCATACGCA ACCACTACCG CCACCCCGAA GAACACGGTT 1301 CCGACCGCTG GTTCAACGCC TTGGGCAGCC GCCGCTTCAG CCGCAACGCC 1351 TGCGTCGTCG TCAGTTGCGG CACGGCGGTA ACGGTTGACG CGCTCACCGA 1401 TGACGGACAT TATCTCGGGG GAACCATCAT GCCCGGTTTC CACCTGATGA 1451 AAGAATCGCT CGCCGTCCGA ACCGCCAACC TCAACCGGCA CGCCGGTAAG 1501 CGTTATCCTT TCCCGACCAC AACGGGCAAT GCCGTCGCCA GCGGCATGAT 1551 GGATGCGGTT TGCGGCTCGG TTATGATGAT GCACGGGCGT TTGAAAGAAA 1601 AAACCGGGGC GGGCAAGCCT GTCGATGTCA TCATTACCGG CGGCGGCGCG 1651 GCAAAAGTTG CCGAAGCCCT GCCGCCTGCA TTTTTGGCGG AAAATACCGT 1701 GCGCGTGGCG GACAACCTCG TCATTCACGG GCTGCTGAAC CTGATTGCCG 1751 CCGAAGGCGG GGAATCGGAA CATACTTAA This encodes a protein having amino acid sequence <SEQ ID 236>: 1 MTVLKPSHWR VLAELADGLP QHVSQLARMA DMKPQQLNGF WQQMPAHIRG 51 LLRQHDGYWR LVRPLAVFDA EGLRELGERS GFQTALKHEC ASSNDEILEL 101 ARIAPDKAHK TICVTHLQSK GRGRQGRKWS HRLGECLMFS FGWVFDRPQY 151 ELGSLSPVAA VACRRALSRL GLKTQIKWPN DLWGRDKLG GILIETVRTG 201 GKTVAWGIG INFVLPKEVE NAASVQSLFQ TASRRGNADA AVLLETLLAE 251 LDAVLLQYAR DGFAPFVAEY QAANRDHGKA VLLLRDGETV FEGTVKGVDG 301 QGVLHLETAE GKQTWSGEI SLRSDDRPVS VPKRRDSERF LLLDGGNSRL 351 KWAWVENGTF ATVGΞAPYRD LSPLGAEWAE KVDGNVRIVG CAVCGEFKKA 401 QVQEQLARKI EWLPSSAQAL GIRNHYRHPE EHGSDRWFNA LGSRRFSRNA 451 CVWSCGTAV TVDALTDDGH YLGGTIMPGF HLMKESLAVR TANLNRHAGK 501 RYPFPTTTGN AVASGMMDAV CGSVMMMHGR LKEKTGAGKP VDVIITGGGA 551 AKVAEALPPA FLAENTVRVA DNLVIHGLLN LIAAEGGESE HT* ORFόla and ORF61-1 show 98.5% identity in 591 aa overlap: 10 20 30 40 50 60 orf61a. pep MTVLKPSHWRVLAELADGLPQHVSQLARMADMKPQQLNGFWQQMPAHIRGLLRQHDGYWR I I M I I I I II I II I II II I II II I I II II I II I II I I I II II II I I I I I I II II I I I I I orf61-1 MTVLKLSHWRVLAELADGLPQHVSQLARMADMKPQQLNGFWQQMPAHIRGLLRQHDGYWR 10 20 30 40 50 60 70 80 90 100 110 120 orf61a .pep LVRPLAVFDAEGLRELGERSGFQTALKHECASSNDEILELARIAPDKAHKTICVTHLQSK I II I II I II I I II I II I II I II I I I II I II I II I II II I I I I I II I I I I I I II II I II I I orf61-1 LVRPLAVFDAEGLRELGERSGFQTALKHECASSNDEILELARIAPDKAHKTICVTHLQSK 70 80 90 100 110 120 130 140 150 160 170 180 orfδla.pep GRGRQGRKWSHRLGECLMFSFGWVFDRPQYELGSLSPVAAVACRRALSRLGLKTQIKWPN II I I II II II II I II II I I I I II I II I II II I I I II I II II II I I I I II II I : II I I II orf61-1 GRGRQGRKWSHRLGECLMFSFGWVFDRPQYELGSLSPVAAVACRRALSRLGLDVQIKWPN 130 140 150 160 170 180 190 200 210 220 230 240 orf61a. pep DLVVGRDKLGGILIETVRTGGKTVAWGIGINFVLPKEVENAASVQSLFQTASRRGNADA II I II I II II II I I I II I I II I II II I II II II I I II I II I I I I I I I I I II II I I I I II I orf61-1 DLWGRDKLGGILIETVRTGGKTVAWGIGINFVLPKEVENAASVQSLFQTASRRGNADA 190 200 210 220 230 240 250 260 270 280 290 300 orf61a . pep AVLLETLLAELDAVLLQYARDGFAPFVAEYQAANRDHGKAVLLLRDGETVFEGTVKGVDG II II I II I: II I I I I I I I II I I II I II II I I I II I II I II II II I II I II II II I I I I I I orf61-1 AVLLETLLVELDAVLLQYARDGFAPFVAEYQAANRDHGKAVLLLRDGETVFEGTVKGVDG 250 260 270 280 290 300 310 320 330 340 350 360 orf61a . pep QGVLHLETAEGKQTWSGEISLRSDDRPVSVPKRRDSERFLLLDGGNSRLKWAWVENGTF I I II I II I j II II II I I I I II I II I II I II II I II I II I II II II I II I I II I I I I I I II orf61-1 QGVLHLETAEGKQTWSGEISLRSDDRPVSVPKRRDSERFLLLDGGNSRLKWAWVENGTF 310 320 330 340 350 360 370 380 390 400 410 420 orf61a. pep ATVGSAPYRDLSPLGAEWAEKVDGNVRIVGCAVCGEFKKAQVQEQLARKIEWLPSSAQAL I II I I I I II I I II II II I I II : II I I II II II I I II II I II II II I II I II I II II II I I orf61-1 ATVGSAPYRDLSPLGAEWAEKADGNVRIVGCAVCGEFKKAQVQEQLARKIEWLPSSAQAL 370 380 390 400 410 420 430 440 450 460 470 480 orf61a. pep GIRNHYRHPEEHGSDRWFNALGSRRFSRNACVWSCGTAVTVDALTDDGHYLGGTIMPGF I II II I II II I I II II I I II I I II I I II II II I I II I II II I II I II I II II II I I I I I I orf61-1 GIRNHYRHPEEHGSDRWFNALGSRRFSRNACVWSCGTAVTVDALTDDGHYLGGTIMPGF 430 440 450 460 470 480 490 500 510 520 530 540 orf61a . pep HLMKESLAVRTANLNRHAGKRYPFPTTTGNAVASGMMDAVCGSVMMMHGRLKEKTGAGKP II I II I II II II II I I II I I I I II I I II II II I I II I II II I II I II II I I I I II II II I orf61-1 HLMKESLAVRTANLNRHAGKRYPFPTTTGNAVASGMMDAVCGSVMMMHGRLKEKTGAGKP 490 500 510 520 530 540 550 560 570 580 590 orf61a . pep VDVIITGGGAAKVAEALPPAFLAENTVRVADNLVIHGLLNLIAAEGGESEHTX I I II I II I II II I I II II II I I II I I I I II II I I I : I II I : II II I I II orf61-l VDVIITGGGAAKVAEALPPAFLAENTVRVADNLVIYGLLNMIAAEGREYEHIX 550 560 570 580 590 Homology with a predicted ORF from N.sonorrhoeae ORFόl shows 94.2% identity over a 189aa overlap with a predicted ORF (ORFόl.ng) from N gonorrhoeae: orf61. ep EISLRSDXRPVSVXKRRDSERFLLLDGGΝS 30 II I II I I III II II I I II II: II II orfδlng TVCEGTVKGVDGRGVLHLETAEGEQTWSGEISLRPDΝRSVSVPKRPDSERFLLLEGGΝS 211 orf61. pep RLKWAWVEΝGTFATVGSAPYRDLSPLGAEWAEKADGΝVRIVGCAVCGEFKKAQVQEQLAR 90 II I I I II I I II II I II II II I I II I I II I II II I II I I II II I I I I II I II IM I I I I I orf61ng RLKWAWVEΝGTFATVGSAPYRDLSPLGAEWAEKADGΝVRIVGCAVCGESKKAQVKEQLAR 271 orf61.pep KIEWLPSSAQAXGIRΝHYRHPEEHGSDRWFΝALGSRRFSRΝACVWSCGTAVTVDALTDD 150 I I II I I II II I II I II II I II I I II I I II II I II I II I II II II I I II I I II I II I I II orfδlng KIEWLPSSAQALGIRΝHYRHPEEHGSDRWFΝALGSRRFSRΝACVWSCGTAVTVDALTDD 331 orf61. pep GHYLGXGTIMPGFHLMKESLAVRTAΝLΝRHAGKRYPFPT 189 I I I II I II II II II I II II I I II I I II I I I I I I II I I orfδlng GHYLG-GTIMPGFHLMKESLAVRTAΝLΝRPAGKRYPFPTTTGΝAVASGMMDAVCGSIMMM 390 An ORFόl ng nucleotide sequence <SEQ LD 237> was predicted to encode a protein having amino acid sequence <SEQ ID 238>: 1 ΛfFSFGWAFDR PQYELGSLSP VAALACRRAL GCLGLETQIK WPNDLWGRD 51 KLGGILIETV RAGGKTVAW GIGINFVLPK EVENAASVQS LFQTASRRGN 101 ADAAVLLETL LAELGAVLEQ YAEEGFAPFL NEYETANRDH GKAVLLLRDG 151 ETVCEGTVKG VDGRGVLHLE TAEGEQTWS GEISLRPDNR SVSVPKRPDS 201 ERFLLLEGGN SRLKWAWVEN GTFATVGSAP YRDLSPLGAE WAEKADGNVR 251 IVGCAVCGES KKAQVKEQLA RKIEWLPSSA QALGIRNHYR HPEEHGSDRW 301 FNALGSRRFS RNACV SCG TAVTVDALTD DGHYLGGTIM PGFHLMKESL 351 AVRTANLNRP AGKRYPFPTT TGNAVASGMM DAVCGSIMMM HGRLKEKNGA 401 GKPVDVIITG GGAAKVAEAL PPAFLAENTV RVADNLVIHG LLNLIAAEGG 451 ESEHA* Further analysis revealed the complete gonococcal DNA sequence <SEQ ID 239> to be: 1 ATGACGGTTT TGAAGCCTTC GCATTGGCGG GTGTTGGCGG AGCTTGCCGA 51 CGGTTTGCCG CAACACGTAT CGCAATTGGC GCGTGAGGCG GACATGAAGC 101 CGCAGCAGCT CAACGGTTTT TGGCAGCAGA TGCCGGCGCA TATACGCGGG 151 CTGTTGCGCC AACACGACGG CTATTGGCGG CTGGTGCGCC CCTTGGCGGT 201 TTTCGATGCC GAAGGTTTGC GCGATCTGGG GGAAAGGTCG GGTTTTCAGA 251 CGGCATTGAA GCACGAGTGC GCGTCCAGCA ACGACGAGAT ACTGGAATTG 301 GCGCGGATTG CGCCGGACAA GGCGCACAAA ACCATATGCG TGACCCACCT 351 GCAAAGTAAG GGCAGGGGGC GGCAGGGGCG GAAGTGGTCG CACCGTTTGG 401 GCGAGTGCCT GATGTTCAGT TTCGGCTGGG CGTTTGACCG GCCGCAGTAT 451 GAGTTGGGTT CGCTGTCGCC TGTTGCGGCA CTTGCGTGCC GGCGCGCTTT 501 GGGGTGTTTG GGTTTGGAAA CGCAAATCAA GTGGCCAAAC GATTTGGTCG 551 TCGGACGCGA CAAATTGGGC GGCATTCTGA TTGAAACAGT CAGGGCGGGC 601 GGTAAAACGG TTGCCGTGGT CGGTATCGGC ATCAATTTCG TGCTGCCCAA 651 GGAAGTGGAA AACGCCGCTT CCGTGCAGTC GCTGTTTCAG ACGGCATCGC 701 GGCGGGGCAA TGCCGATGCC GCCGTATTGC TGGAAACATT GCTTGCGGAA 751 CTGGGCGCGG TGTTGGAACA ATATGCGGAA GAAGGGTTCG CGCCATTTTT 801 AAATGAGTAT GAAACGGCCA ACCGCGACCA CGGCAAGGCG GTATTGCTGT 851 TGCGCGACGG CGAAACCGTG TGCGAAGGCA CGGTTAAAGG CGTGGACGGA 901 CGAGGCGTTC TGCACTTGGA AACGGCAgaa ggcgaACAGa cggtcgtcag 951 cggcgaaaTC AGcctGCggc ccgacaacaG GTCGGtttcc gtgccgaagc 1001 ggccggatTC GgaacgtTTT tTGCtgttgg aaggcgggaa cagccgGCTC 1051 AAGTGGGCGT GggtggAAAa cggcacgttc gcaaccgtgg gcagcgcgCc 1101 gtaCCGCGAT TTGTCGCCTT TGGGCGCGGA GTGGGCGGAA AAGGCGGATG 1151 GAAATGTCCG CATCGTCGGT TGCGCCGTGT GCGGAGAATC CAAAAAGGCA 1201 CAAGTGAAGG AACAGCTCGC CCGAAAAATC GAGTGGCTGC CGTCTTCCGC 1251 ACAGGCTTTG GGCATACGCA ACCACTACCG CCACCCCGAA GAACACGGTT 1301 CCGACCGTTG GTTCAACGCC TTGGGCAGCC GCCGCTTCAG CCGCAACGCC 1351 TGCGTCGTCG TCAGTTGCGG CACGGCGGTA ACGGTTGACG CGCTCACCGA 1401 TGACGGACAT TATCTCGGCG GAACCATCAT GCCCGGCTTC CACCTGATGA 1451 AAGAATCGCT CGCCGTCCGA ACCGCCAACC TCAACCGCCC CGCCGGCAAA 1501 CGTTACCCTT TCCCGACCAC AACGGGCAAC GCCGTCGCAA GCGGCATGAT 1551 GGACGCGGTT TGCGGCTCGA TAATGATGAT GCACGGCCGT TTGAAAGAAA 1601 AAAACGGCGC GGGCAAGCCT GTCGATGTCA TCATTACCGG CGGCGGCGCG 1651 GCGAAAGTCG CCGAAGCCCT GCCGCCTGCA TTTTTGGCGG AAAATACCGT 1701 GCGCGTGGCG GACAACCTCG TCATCCACGG GCTGCTGAAC CTGATTGCCG 1751 CCGAAGGCGG GGAATCGGAA CACGCTTAA This corresponds to the amino acid sequence <SEQ ID 240; ORF61ng-l>: 1 MTVLKPSHWR VLAELADGLP QHVSQLAREA DMKPQQLNGF WQQMPAHIRG 51 LLRQHDGYWR LVRPLAVFDA EGLRDLGERS GFQTALKHEC ASSNDEILEL 101 ARIAPDKAHK TICVTHLQSK GRGRQGRKWS HRLGECLMFS FGWAFDRPQY 151 ELGSLSPVAA LACRRALGCL GLETQIKWPN DLWGRDKLG GILIETVRAG 201 GKTVAWGIG INFVLPKEVE NAASVQSLFQ TASRRGNADA AVLLETLLAE 251 LGAVLEQYAE EGFAPFLNEY ETANRDHGKA VLLLRDGETV CEGTVKGVDG 301 RGVLHLETAE GEQT SGEI SLRPDNRSVS VPKRPDSERF LLLEGGNSRL 351 KWAWVENGTF ATVGSAPYRD LSPLGAEWAE KADGNVRIVG CAVCGESKKA 401 QVKEQLARKI EWLPSSAQAL GIRNHYRHPE EHGSDRWFNA LGSRRFSRNA 451 CVWSCGTAV TVDALTDDGH YLGGTIMPGF HLMKESLAVR TANLNRPAGK 501 RYPFPTTTGN AVASGMMDAV CGSIMMMHGR LKEKNGAGKP VDVIITGGGA 551 AKVAEALPPA FLAENTVRVA DNLVIHGLLN LIAAEGGESE HA* ORF61ng-l and ORF61-1 show 93.9% identity in 591 aa overlap: orf 61ng-l . pep MTVLKPSHWRVLAELADGLPQHVSQLAREADMKPQQLNGFWQQMPAHIRGLLRQHDGYWR 60 II I II II II II II II II I I II I II I II I I II I II I I II I I I I I I I I I I II I I I I II I I orf61-l MTVLKLSHWRVLAELADGLPQHVSQLARMADMKPQQLNGFWQQMPAHIRGLLRQHDGYWR 60 orf 61ng-l . pep LVRPLAVFDAEGLRDLGERSGFQTALKHECASSNDEILELARIAPDKAHKTICVTHLQSK 120 I I I II I II I I II II : I II I I I I I I II II II II II I II II I II II I I II II I I II II I I I I orf61-l LVRPLAVFDAEGLRELGERSGFQTALKHECASSNDEILELARIAPDKAHKTICVTHLQSK 120 orf 61ng-l . pep GRGRQGRKWSHRLGECLMFSFGWAFDRPQYELGSLSPVAALACRRALGCLGLETQIKWPN 180 I II II II II II II II II I II I I I : I II II II II II I I I II : II II I I : I II :: I I I II I orf61-l GRGRQGRKWSHRLGECLMFSFGWVFDRPQYELGSLSPVAAVACRRALSRLGLDVQIKWPN 180 orf 61ng-l . pep DLWGRDKLGGILIETVRAGGKTVAWGIGINFVLPKEVENAASVQSLFQTASRRGNADA 240 I II I I I I II II II II I I I : I II II I II I I I I I I I II II I I I II I I II I I II I II II I I II orf61-l DLVVGRDKLGGILIETVRTGGKTVAWGIGINFVLPKEVENAASVQSLFQTASRRGNADA 240 orf 61ng-l . pep AVLLETLLAELGAVLEQYAEEGFAPFLNEYETANRDHGKAVLLLRDGETVCEGTVKGVDG 300 I II I I I I I : II III I I I : : I I II I : II :: II I II II I I I II II I I II I I I I I II I I orf61-l AVLLETLLVELDAVLLQYARDGFAPFVAEYQAANRDHGKAVLLLRDGETVFEGTVKGVDG 300 orf 61ng-l . pep RGVLHLETAEGEQTWSGEISLRPDNRSVSVPKRPDSERFLLLEGGNSRLKWAWVENGTF 360 : II I I I I II I I: I I I I II I I I II M I II II I I I II II I II : I II I II I I II II I I II orf61-l QGVLHLETAEGKQTWSGEISLRSDDRPVSVPKRRDSERFLLLDGGNSRLKWAWVENGTF 360 orf 61ng-l . pep ATVGSAPYRDLSPLGAEWAEKADGNVRIVGCAVCGESKKAQVKEQLARKIEWLPSSAQAL 420 I I I I I I I II II II II II I I I I I I I II I II I I II II I I II I I : II II I I II I II II I I II orf61-l ATVGSAPYRDLSPLGAEWAEKADGNVRIVGCAVCGEFKKAQVQEQLARKIEWLPSSAQAL 420 orf 61ng-l . pep GIRNHYRHPEEHGSDRWFNALGΞRRFSRNACWVSCGTAVTVDALTDDGHYLGGTIMPGF 480 I II I I II II I I I II II II I I I II I I II I I II I I I I I I I I I II II II I I I I II I I I I II I I orf61-l GIRNHYRHPEEHGSDRWFNALGSRRFSRNACVWSCGTAVTVDALTDDGHYLGGTIMPGF 480 orf 61ng-l . pep HLMKESLAVRTANLNRPAGKRYPFPTTTGNAVASGMMDAVCGSIMMMHGRLKEKNGAGKP 540 I II I I II II I I I II II II I I I II I I I I I II I II I I I II I I I I: I I I I I I I I I I: I I I I I orf61-l HLMKESLAVRTANLNRHAGKRYPFPTTTGNAVASGMMDAVCGSVMMMHGRLKEKTGAGKP 540 orfδlng-l.pep VDVIITGGGAAKVAEALPPAFLAENTVRVADNLVIHGLLNLIAAEGGESEHAX 593 I II I I II I II II I I I II I II II I II I II II II I II : II II : II II I I II orf61-l VDVIITGGGAAKVAEALPPAFLAENTVRVADNLVIYGLLNMIAAEGREYEHIX 593 Based on this .analysis, including the homology with the baf protein of B.pertussis .and the presence of a putative prokaryotic membrane lipoprotein lipid attachment site, it is predicted that these proteins from N. meningitidis .and N.gonorrhoeae, and their epitopes, could be useful antigens for vaccines or diagnostics, or for raising antibodies. Example 29 The following partial DNA sequence was identified in N. meningitidis <SEQ ID 241>: 1 ATGTTTTACC AAATCCTTGC CCTGATTATC TGGAGCAGCT CGTTTATTGC 51 CGCCAAATAT GTCTATGGCG GCATCGATCC CGCATTGATG GTCGGCGTGC 101 GCCTGCTAAT TGCCGCGCTG CCTGCACTGC CCGCCTGCCG CCGTCATGTC 151 GGCAAGATTC CGCGTGAGGA ATGGAAGCCG TTGCTGATTG TGTCGTTCGT 201 CAACTATGTG CTGACCCTGC TGCTTCAGTT TGTCGGGTTG AAATACACTT 251 CCGCCGCCAG CGCATCGGTC ATTGTCGGAC TCGAGCCGCT GCTGATGGTG 301 TTTGTCGGAC ACTTTTTCTT CAACGACAAA GCGCGTGCCT ACCACTGGAT 351 ATGCGGCGCG GCGGCATTTG CCGGTGTCGC GCTGCTGATG GCGGGCGGTG 401 CGGaAGAGGG CGGCGaAGTC GGCTGGTTCG GCTGCCTGCT GGTGTTGTTG 451 GCGGGCGCGG GCTTTTGTGC CGCTATGCGT CCGACGCAAA GGCTGATTGC 501 ACGCATCGGC GCACCGGCAT TCACATCTGT TTCCATTGCC GCCGCATCGT 551 TGATGTGCCT GCCGTTTTCG CTTGCTTTGG CGCAAAGTTA TACCGTGGAC 601 TGGAGCGTCG GGATGGTATT GTCGCTGCTG TATTTGGGTT TGGGGTGC .. This corresponds to the amino acid sequence <SEQ ID 242; ORF62>: 1 MFYQILALII WSSSFIAAKY VYGGIDPALM VGVRLLIAAL PALPACRRHV 51 GKIPREEWKP LLIVSFVNYV LTLLLQFVGL KYTSAASASV IVGLEPLLMV 101 FVGHFFFNDK ARAYHWICGA AAFAGVALLM AGGAEEGGEV GWFGCLLVLL 151 AGAGFCAAMR PTQRLIARIG APAFTSVSIA AASLMCLPFS LALAQSYTVD 201 WSVGMVLSLL YLGLGC.. Further work revealed the complete nucleotide sequence <SEQ ID 243>: 1 ATGTTTTACC AAATCCTTGC CCTGATTATC TGGAGCAGCT CGTTTATTGC 51 CGCCAAATAT GTCTATGGCG GCATCGATCC CGCATTGATG GTCGGCGTGC 101 GCCTGCTAAT TGCCGCGCTG CCTGCACTGC CCGCCTGCCG CCGTCATGTC 151 GGCAAGATTC CGCGTGAGGA ATGGAAGCCG TTGCTGATTG TGTCGTTCGT 201 CAACTATGTG CTGACCCTGC TGCTTCAGTT TGTCGGGTTG AAATACACTT 251 CCGCCGCCAG CGCATCGGTC ATTGTCGGAC TCGAGCCGCT GCTGATGGTG 301 TTTGTCGGAC ACTTTTTCTT CAACGACAAA GCGCGTGCCT ACCACTGGAT 351 ATGCGGCGCG GCGGCATTTG CCGGTGTCGC GCTGCTGATG GCGGGCGGTG 401 CGGAAGAGGG CGGCGAAGTC GGCTGGTTCG GCTGCCTGCT GGTGTTGTTG 451 GCGGGCGCGG GCTTTTGTGC CGCTATGCGT CCGACGCAAA GGCTGATTGC 501 ACGCATCGGC GCACCGGCAT TCACATCTGT TTCCATTGCC GCCGCATCGT 551 TGATGTGCCT GCCGTTTTCG CTTGCTTTGG CGCAAAGTTA TACCGTGGAC 601 TGGAGCGTCG GGATGGTATT GTCGCTGCTG TATTTGGGTT TGGGGTGCGG 651 CTGGTACGCC TATTGGCTGT GGAACAAGGG GATGAGCCGT GTTCCTGCCA 701 ATGTTTCGGG ACTGTTGATT TCGCTCGAAC CCGTCGTCGG CGTGCTGCTG 751 GCGGTTTTGA TTTTGGGCGA ACACCTGTCG CCCGTGTCCG CCTTGGGCGT 801 GTTTGTCGTC ATCGCCGCCA CCTTGGTTGC CGGCCGGCTG TCGCATCAAA 851 AATAA This corresponds to the .amino acid sequence <SEQ ID 244; ORF62-l>: 1 MFYQILALII WSSSFIAAKY VYGGIDPALM VGVRLLIAAL PALPACRRHV 51 GKIPREEWKP LLIVSFVNYV LTLLLQFVGL KYTSAASASV IVGLEPLLMV 101 FVGHFFFNDK ARAYHWICGA AAFAGVALLM AGGAEEGGEV GWFGCLLVLL 151 AGAGFCAAMR PTQRLIARIG APAFTSVSIA AASLMCLPFS LALAQSYTVD 201 WSVGMVLSLL YLGLGCGWYA YWLWNKGMSR VPANVSGLLI SLEPWGVLL 251 AVLILGEHLS PVSALGVFW IAATLVAGRL SHQK* Computer analysis of this amino acid sequence gave the following results: Homology with hypothetical transmembrane protein HI0976 of H. influenzae (accession number 057147) ORF62 and HI0976 show 50% aa identity in 114aa overlap: Orf62 1 MFYQILALIIWSSSFIAAKYVYGGIDPALMVGVRXXXXXXXXXXXCRRHVGKIPREEWKP 60 M YQILAL+IWSSS I K Y +DP L+V VR R KI + K HI0976 1 MLYQILALLIWSSSLIVGKLTYSMMDPVLWQVRLIIAMIIVMPLFLRRWKKIDKPMRKQ 60 0rf62 61 LLIVSFVNYVLTLLLQFVGLKYTSAASASVIVGLEPLLMVFVGHFFFNDKARAY 114 L ++F NY LLQF+GLKYTSA+SA ++GLEPLL+VFVGHFFF K + HI0976 61 LWWLAFFNYTAVFLLQFIGLKYTSASSAVTMIGLEPLLWFVGHFFFKTKQNGF 114 Homology with a predicted ORF from N.meninsitidis (strain A) ORF62 shows 99.5% identity over a 216aa overlap with an ORF (ORF62a) from strain A of N meningitidis: 10 20 30 40 50 60 orf 62 . pep MFYQILALIIWSSSFIAAKYVYGGIDPALMVGVRLLIAALPALPACRRHVGKIPREEWKP I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I orf 62a MFYQILALIIWSSSFIAAKYVYGGIDPALMVGVRLLIAALPALPACRRHVGKIPREEWKP 10 20 30 40 50 60 70 80 90 100 110 120 orf 62 . pep LLIVSFVΝYVLTLLLQFVGLKYTSAASASVIVGLEPLLMVFVGHFFFΝDKARAYHWICGA II II I I I II I II I I II I II II I I II I II II II I II I II II I I II II I II I I II I I I I I II orf 62a LLIVSFVΝYVLTLLLQFVGLKYTSAASASVIVGLEPLLMVFVGHFFFΝDKARAYHWICGA 70 30 90 100 110 120 130 140 150 160 170 180 orf 62 . pep AAFAGVALLMAGGAEEGGEVGWFGCLLVLLAGAGFCAAMRPTQRLIARIGAPAFTSVSIA I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I orf62a AAFAGVALLMAGGAEEGGEVGWFGCLLVLLAGAGFCAAMRPTQRLIARIGAPAFTSVSIA 130 140 150 160 170 180 190 200 210 orf 62 . pep AASLMCLPFSLALAQSYTVDWSVGMVLSLLYLGLGC I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I : I I orf62a AASLMCLPFSLALAQSYTVDWSVGMVLSLLYLGVGCSWYAYWLWNKGMSRVPANVSGLLI 190 200 210 220 230 240 orf62a SLEPWGVLLAVLILGEHLSPVSVLGVFWIAATLVAGRLSHQKX 250 226600 270 280 The complete length ORF62a nucleotide sequence <SEQ ID 245> is 1 ATGTTTTACC AAATCCTTGC CCTGATTATC TGGAGCAGCT CGTTTATTGC 51 CGCCAAATAT GTCTATGGCG GCATCGATCC CGCATTGATG GTCGGCGTGC 101 GCCTGCTGAT TGCTGCGCTG CCTGCACTGC CCGCCTGCCG CCGTCATGTC 151 GGCAAGATTC CGCGTGAGGA ATGGAAGCCG TTGCTGATTG TGTCGTTCGT 201 CAACTATGTG CTGACCCTGC TACTTCAGTT TGTCGGGTTG AAATACACTT 251 CCGCCGCCAG CGCATCGGTC ATTGTCGGAC TCGAGCCACT GCTGATGGTG 301 TTTGTCGGAC ACTTTTTCTT CAACGACAAA GCGCGTGCCT ACCACTGGAT 351 ATGCGGCGCG GCGGCATTTG CCGGTGTCGC GCTGCTGATG GCGGGCGGTG 401 CGGAAGAGGG CGGCGAAGTC GGCTGGTTCG GCTGCCTGCT GGTGTTGTTG 451 GCGGGCGCGG GCTTTTGTGC CGCTATGCGT CCGACGCAAA GGCTGATTGC 501 ACGCATCGGC GCACCGGCAT TCACATCTGT TTCCATTGCC GCCGCATCGT 551 TGATGTGCCT GCCGTTTTCG CTTGCTTTGG CGCAAAGTTA TACCGTGGAC 601 TGGAGCGTCG GAATGGTATT GTCGCTGCTG TATTTGGGCG TGGGGTGCAG 651 CTGGTACGCC TATTGGCTGT GGAACAAGGG GATGAGCCGT GTTCCTGCCA 701 ACGTTTCGGG ACTGTTGATT TCGCTCGAAC CCGTCGTCGG CGTGCTGCTG 751 GCGGTTTTGA TTTTGGGCGA ACACCTGTCG CCCGTGTCCG TCTTGGGCGT 801 GTTTGTCGTC ATCGCCGCCA CCTTGGTTGC CGGCCGGCTG TCGCATCAAA 851 AATAA This encodes a protein having amino acid sequence <SEQ ID 246>: 1 MFYQILALII WSSSFIAAKY VYGGIDPALM VGVRLLIAAL PALPACRRHV 51 GKIPREEWKP LLIVSFVNYV LTLLLQFVGL KYTSAASASV IVGLEPLLMV 101 FVGHFFFNDK ARAYHWICGA AAFAGVALLM AGGAEEGGEV GWFGCLLVLL 151 AGAGFCAAMR PTQRLIARIG APAFTSVSIA AASLMCLPFS LALAQSYTVD 201 WSVGMVLSLL YLGVGCSWYA YWLWNKGMSR VPANVSGLLI SLEPWGVLL 251 AVLILGEHLS PVSVLGVF IAATLVAGRL SHQK* ORF62a and ORF62-1 show 98.9% identity in 284 aa overlap: orf 62a . pep MFYQILALIIWSSSFIAAKYVYGGIDPALMVGVRLLIAALPALPACRRHVGKIPREEWKP 60 I I II I I I I II I I II II I I II I I I I I II I II I I I II II I I II II I II I I I I I I II I I I I I I orf62-1 MFYQILALIIWSSSFIAAKYVYGGIDPALMVGVRLLIAALPALPACRRHVGKIPREEWKP 60 orf62a . pep LLIVSFVNYVLTLLLQFVGLKYTSAASASVIVGLEPLLMVFVGHFFFNDKARAYHWICGA 120 II I I II II I I II II I II I I I II II II I II I I I II I I I I II II II I II I I I II II I II I II orf62-l LLIVSFVNYVLTLLLQFVGLKYTSAASASVIVGLEPLLMVFVGHFFFNDKARAYHWICGA 120 orf62a . pep AAFAGVALLMAGGAEEGGEVGWFGCLLVLLAGAGFCAAMRPTQRLIARIGAPAFTSVSIA 180 II II I I I I II I I II I I II I II I I II I II I II II I II I I II I I I II II I I I I I I II I II I I orf62-1 AAFAGVALLMAGGAEEGGEVGWFGCLLVLLAGAGFCAAMRPTQRLIARIGAPAFTSVSIA 180 orf62a. pep AASLMCLPFSLALAQSYTVDWSVGMVLSLLYLGVGCSWYAYWLWNKGMSRVPANVSGLLI 240 I I I II I I I I I II I II II I I I II I I II I I II II I: I I : I I I II I I II I I I I I I II I I I I I I orf62-1 AASLMCLPFSLALAQSYTVDWSVGMVLSLLYLGLGCGWYAYWLWNKGMSRVPANVSGLLI 240 orf62a.pep SLEPWGVLLAVLILGEHLSPVSVLGVFWIAATLVAGRLSHQKX 285 I I I I II II I I I II II I II I II I I: I I II I II II I II II II I I I I I orf62-l SLEPWGVLLAVLILGEHLSPVSALGVFWIAATLVAGRLSHQKX 285 Homology with a predicted ORF from N.sonorrhoeae ORF62 shows 99.5% identity over a 216aa overlap with a predicted ORF (ORF62.ng) from N. gonorrhoeae: orf62.pep MFYQILALIIWSSSFIAAKYVYGGIDPALMVGVRLLIAALPALPACRRHVGKIPREEWKP 60 I I II I II II II : II II I II I II I I I I II I II II I II I II II I I I I I I I I II I I I I I I II I orf62ng MFYQILALIIWGSSFIAAKYVYGGIDPALMVGVRLLIAALPALPACRRHVGKIPREEWKP 60 orf62.pep LLIVSFVNYVLTLLLQFVGLKYTSAASASVIVGLEPLLMVFVGHFFFNDKARAYHWICGA 120 I I II II II II II II I II II I I II I I I II I II I I I II I II II I I I II I II I I I II I I I I II orf62ng LLIVSFVNYVLTLLLQFVGLKYTSAASASVIVGLEPLLMVFVGHFFFNDKARAYHWICGA 120 orf62.pep AAFAGVALLMAGGAEEGGEVGWFGCLLVLLAGAGFCAAMRPTQRLIARIGAPAFTSVSIA 180 II I II II I II II I I I I I I I II I I II I I I I II II I II I II I I I II II I II II I II I I I I I I orf62ng AAFAGVALLMAGGAEEGGEVGWFGCLLVLLAGAGFCAAMRPTQRLIARIGAPAFTSVSIA 180 orf62. pep AASLMCLPFSLALAQSYTVDWSVGMVLSLLYLGLGC 216 I I II I II II II I I II I I I I II I I II I II II I II I I I orf62ng AASLMCLPFSLALAQSYTVDWSVGMVLSLLYLGLGCGWYAYWLWNKGMSRVPANASGLLI 240 The complete length ORF62ng nucleotide sequence <SEQ LD 247> is: 1 ATGTTTTACC AAATCCTTGC CCTGATTATC TGGGGCAGCT CGTTTATTGC 51 CGCCAAATAT GTCTATGGCG GCATCGATCC CGCATTGATG GTCGGCGTGC 101 GCCTGCTGAT TGCCGCGCTG CCTGCACTGC CCGCCTGCCG CCGTCATGTC 151 GGCAAGATTC CGCGTGAGGA ATGGAAGCCG TTGCTGATTG TGTCGTTCGT 201 CAACTATGTG CTGACCCTGC TGCTTCAGTT TGTCGGGTTG AAATACACTT 251 CCGCCGCCAG CGCATCGGTC ATTGTCGGAC TCGAGCCGCT GCTGATGGTG 301 TTTGTCGGAC ACTTTTTCTT CAACGACAAA GCGCGTGCCT ACCACTGGAT 351 ATGCGGCGCG GCGGCATTTG CCGGTGTCGC GCTGCTGATG GCGGGCGGTG 401 CGGAAGAGGG CGGCGAAGTC GGCTGGTTCG GCTGCCTGCT GGTGTTGTTG 451 GCGGGCGCGG GCTTTTGTGC CGCTATGCGT CCGACGCAAA GGCTGATTGC 501 CCGCATCGGC GCACCGGCAT TCACATCTGT TTCCATTGCC GCCGCATCGT 551 TGATGTGCCT GCCGTTTTCG CTTGCTTTGG CGCAAAGTTA TACCGTGGAC 601 TGGAGCGTCG GGATGGTATT GTCGCTGTTG TATTTGGGTT TGGGGTGCGG 651 CTGGTACGCC TATTGGCTGT GGAACAAGGG GATGAGCCGT GTTCCTGCCA 701 ACGCGTCGGG ACTGTTGATT TCGCTCGAAC CCGTCGTCGG CGTGCTGTTG 751 GCGGTTTTGA TTTTGGGCGA ACATTTATCG CCCGTGTCCG CCTTGGGCGT 801 GTTTGTCGTC ATCGCCGCCA CTTTCGCCGC CGGCCGGCTG TCGCGCAGGG 851 ACGCGCAAAA CGGCAATGCC GTCTGA This encodes a protein having amino acid sequence <SEQ ID 248>: 1 MFYQILALII WGSSFIAAKY VYGGIDPALM VGVRLLIAAL PALPACRRHV 51 GKIPREEWKP LLIVSFVNYV LTLLLQFVGL KYTSAASASV IVGLEPLLMV 101 FVGHFFFNDK ARAYHWICGA AAFAGVALLM AGGAEEGGEV GWFGCLLVLL 151 AGAGFCAAMR PTQRLIARIG APAFTSVSIA AASLMCLPFS LALAQSYTVD 201 WSVGMVLSLL YLGLGCGWYA YWLWNKGMSR VPANASGLLI SLEPWGVLL 251 AVLILGEHLS PVSALGVF IAATFAAGRL SRRDAQNGNA V* ORF62ng and ORF62-1 show 97.9% identity in 283 aa overlap: 10 20 30 40 50 60 orf62ng.pep MFYQILALIIWGSSFIAAKYVYGGIDPALMVGVRLLIAALPALPACRRHVGKIPREEWKP I II I II II II I : II I II I I I II I I II I II II I I II II II II II I II II II II II I II I I I orf62-1 MFYQILALIIWSSSFIAAKYVYGGIDPALMVGVRLLIAALPALPACRRHVGKIPREEWKP 10 20 30 40 50 60 70 80 90 100 110 120 orf62ng . pep LLIVSFVNYVLTLLLQFVGLKYTSAASASVIVGLEPLLMVFVGHFFFNDKARAYHWICGA II I II I II II II II I II I I II I I II I I II I I I II I I I II II I II II II I I I II I I II I I I orf62-1 LLIVSFVNYVLTLLLQFVGLKYTSAASASVIVGLEPLLMVFVGHFFFNDKARAYHWICGA 70 80 90 100 110 120 130 140 150 160 170 180 orf62ng . pep AAFAGVALLMAGGAEEGGEVGWFGCLLVLLAGAGFCAAMRPTQRLIARIGAPAFTSVSIA II II I II I II I I II I I I II II I I II I II II I II I I II II I I II I I I II II I II I I II I I I orf62-1 AAFAGVALLMAGGAEEGGEVGWFGCLLVLLAGAGFCAAMRPTQRLIARIGAPAFTSVSIA 130 140 150 160 170 180 190 200 210 220 230 240 orf62ng . pep AASLMCLPFSLALAQSYTVDWSVGMVLSLLYLGLGCGWYAYWLWNKGMSRVPANASGLLI I I II I II II II II I II I II I I II II II I I I I II I I II II I I I I I II I II I I II I: II I I I orf62-1 AASLMCLPFSLALAQSYTVDWSVGMVLSLLYLGLGCGWYAYWLWNKGMSRVPANVSGLLI 190 200 210 220 230 240 250 260 270 280 290 orf62ng . pep SLEPWGVLLAVLILGEHLSPVSALGVFWIAATFAAGRLSRRDAQNGNAVX II I II I II II II I I I II I II M I I I II II II II I : I II I I :: orf62-l SLEPWGVLLAVLILGEHLSPVSALGVFWIAATLVAGRLSHQKX 250 260 270 280 Furthermore, ORF62ng shows significant homology to a hypothetical H.influenzae protein: sp|Q57147 |Y976_HAEIN HYPOTHETICAL PROTEIN HI0976 >gi | 1074589 Ipir MB64163 hypothetical protein HI0976 - Haemophilus influenzae (strain Rd KW20) >gi 11574004 (U32778) hypothetical [Haemophilus influenzae] Length = 128 Score = 106 bits (262), Expect = 2e-22 Identities = 56/114 (49%), Positives = 68/114 (59%) Query: 1 MFYQILALIIWGSSFIAAKYVYGGIDPALMVGVRXXXXXXXXXXXCRRHVGKIPREEWKP 60 M YQILAL+IW SS I K Y +DP L+V VR R KI + K Sbjct: 1 MLYQILALLIWSSSLIVGKLTYSMMDPVLWQVRLIIAMIIVMPLFLRRWKKIDKPMRKQ 60 Query: 61 LLIVSFVNYVLTLLLQFVGLKYTSAASASVIVGLEPLLMVFVGHFFFNDKARAY 114 L ++F NY LLQF+GLKYTSA+SA ++GLEPLL+VFVGHFFF K + Sbjct: 61 LWWLAFFNYTAVFLLQFIGLKYTSASSAVTMIGLEPLLWFVGHFFFKTKQNGF 114 Based on this analysis, including the homology with the transmembrane protein of H.influenzae and the putative leader sequecne and several transmembrane domains in the gonococcal protein, it is predicted that these proteins from N. meningitidis and N.gonorrhoeae, and their epitopes, could be useful antigens for vaccines or diagnostics, or for raising antibodies. Example 30 The following partial DNA sequence was identified in N. meningitidis <SEQ ID 249>: 1 ATGCGCCGTT TTCTACCGAT CGCAGCCATA TGCGCmGwms TCCTGkkGTA 51 sGGACTGACG GCGGCAACCG GCAGCACCAG TTCGCTGGCG GATTATTTCT 101 GGTGGATTGT TGCGTTCAGC GCAATGCTGC TGCTGGTGTT GTCCGCCGTT 151 TTGGCACGTT ATGTCATATT GCTGTTGAAA GACAGGCGCG ACGGCGTATT 201 CGGTTCGStA srTyGCCAAA gsGCCTgkks TGGG.ATGTT TACGCTGGTT 251 GCCGkACTGC CCGGCGTGTT TCTGTTCGGC TTTCCCGCAC AGTTCATCAA 301 CGGCACGATT AATTCGTGGT TCGGCAACGA TACCCACGAG GCGCTTGAAC 351 GCAGCCTCAA TTTGAGCAAG TCCGCATTGA ATTTGGCGGC AGACAACGCC 401 CTCGGCAACG CCGTCCCCGT GCAGATAGAC CTCATCGGCG CGGCTTCCCT 451 GCCCGGGGAT ATGGGCAGGG TGCTGGAACA TTACGCCGGC AGCGGTTTTG 501 CCCAGCTTGC CCTGTACAAy ksCGCAAGCG GCAAAATCGA AAAAAGCATC 551 AACCCGCACA AGCTCGATCA GCCGTTTCCA GGTAAGGCGC GTTGGGAaAa 601 AATCCaACGG GCGGGTTCGG TCAGGGATTT GGAAAGCATA GGCGGCGTAT 651 TGTaCGCGCA GGGCTGGCTG TCGGCGGGTA CGCACwACGG GCGCGATTAC 701 GCCTTGTTTT TCCGTCAGCC GGTTCCCAAA GGCGTGGCAG AGGATGCCGT 751 yTTAATCGAA AAGGCAAGGG CGAAATATGC TGAGTTGAGT TACAGCAAAA 801 AAGGTTTGCA GACCTTTTTC CTGGCAACCC TGCTGATTGC CTCGCTGCTG 851 TCGATTTTTC TTGCACTGGT CATGGCACTG TATTTCGCCC GCCGTTTCGT 901 CGAACCCGTC CTATCGCTTG CCGAGGGGGC GAAGGCGGTG GCGCAAGGCG 951 ATTTCAGCCA GACGCGCCCC GTGTTGCGCA ACGACGAGTT CGGACGCTTG 1001 ACCArGTTGT TCAACCACAT GACCGAGCAG CTTTCCATCG CCAAAGATGC 1051 AGACGAGCGC AACCGCCGGC GCGAGGAAGC CGCCAGGCAT TATCTTGAAT 1101 GCGTGTTGGA GGGGCTGACC ACGGGCGTGG TGGTGTTTGA CGAACAAGGC 1151 TGTCTGAAAA CCTTCAACAA AGCGGCGGGT ACC.. This corresponds to the amino acid sequence <SEQ ID 250; ORF64>: 1 MRRFLPIAAI CAXXLXXGLT AATGSTSSLA DYFWWIVAFS AMLLLVLSAV 51 LARYVILLLK DRRDGVFGSX XAKXPXXXMF TLVAXLPGVF LFGFPAQFIN 101 GTINSWFGND THEALERSLN LSKSALNLAA DNALGNAVPV QIDLIGAASL 151 PGDMGRVLEH YAGSGFAQLA LYNXASGKIE KSINPHKLDQ PFPGKARWEK 201 IQRAGSVRDL ESIGGVLYAQ GWLSAGTHXG RDYALFFRQP VPKGVAEDAV 251 LIEKARAKYA ELSYSKKGLQ TFFLATLLIA SLLSIFLALV MALYFARRFV 301 EPVLSLAEGA KAVAQGDFSQ TRPVLRNDEF GRLTXLFNHM TEQLSIAKDA 351 DERNRRREEA ARHYLECVLE GLTTGVWFD EQGCLKTFNK AAGT .. Further work revealed the complete nucleotide sequence <SEQ ID 251>: 1 ATGCGCCGTT TTCTACCGAT CGCAGCCATA TGCGCCGTCG TCCTGTTGTA 51 CGGACTGACG GCGGCAACCG GCAGCACCAG TTCGCTGGCG GATTATTTCT 101 GGTGGATTGT TGCGTTCAGC GCAATGCTGC TGCTGGTGTT GTCCGCCGTT 151 TTGGCACGTT ATGTCATATT GCTGTTGAAA GACAGGCGCG ACGGCGTATT 201 CGGTTCGCAG ATTGCCAAAC GCCTTTCTGG GATGTTTACG CTGGTTGCCG 251 TACTGCCCGG CGTGTTTCTG TTCGGCGTTT CCGCACAGTT CATCAACGGC 301 ACGATTAATT CGTGGTTCGG CAACGATACC CACGAGGCGC TTGAACGCAG 351 CCTCAATTTG AGCAAGTCCG CATTGAATTT GGCGGCAGAC AACGCCCTCG 401 GCAACGCCGT CCCCGTGCAG ATAGACCTCA TCGGCGCGGC TTCCCTGCCC 451 GGGGATATGG GCAGGGTGCT GGAACATTAC GCCGGCAGCG GTTTTGCCCA 501 GCTTGCCCTG TACAATGCCG CAAGCGGCAA AATCGAAAAA AGCATCAACC 551 CGCACAAGCT CGATCAGCCG TTTCCAGGTA AGGCGCGTTG GGAAAAAATC 601 CAACGGGCGG GTTCGGTCAG GGATTTGGAA AGCATAGGCG GCGTATTGTA 651 CGCGCAGGGC TGGCTGTCGG CGGGTACGCA CAACGGGCGC GATTACGCCT 701 TGTTTTTCCG TCAGCCGGTT CCCAAAGGCG TGGCAGAGGA TGCCGTCTTA 751 ATCGAAAAGG CAAGGGCGAA ATATGCTGAG TTGAGTTACA GCAAAAAAGG 801 TTTGCAGACC TTTTTCCTGG CAACCCTGCT GATTGCCTCG CTGCTGTCGA 851 TTTTTCTTGC ACTGGTCATG GCACTGTATT TCGCCCGCCG TTTCGTCGAA 901 CCCGTCCTAT CGCTTGCCGA GGGGGCGAAG GCGGTGGCGC AAGGCGATTT 951 CAGCCAGACG CGCCCCGTGT TGCGCAACGA CGAGTTCGGA CGCTTGACCA 1001 AGTTGTTCAA CCACATGACC GAGCAGCTTT CCATCGCCAA AGAAGCAGAC 1051 GAGCGCAACC GCCGGCGCGA GGAAGCCGCC AGGCATTATC TTGAATGCGT 1101 GTTGGAGGGG CTGACCACGG GCGTGGTGGT GTTTGACGAA CAAGGCTGTC 1151 TGAAAACCTT CAACAAAGCG GCGGAACAGA TTTTGGGGAT GCCGCTTACC 1201 CCCCTGTGGG GCAGCAGCCG GCACGGTTGG CACGGCGTTT CGGCGCAGCA 1251 GTCCCTGCTT GCCGAAGTGT TTGCCGCCAT CGGCGCGGCG GCAGGTACGG 1301 ACAAACCGGT CCATGTGAAA TATGCCGCGC CGGACGATGC CAAAATCCTG 1351 CTGGGCAAGG CAACCGTCCT GCCCGAAGAC AACGGCAACG GCGTGGTAAT 1401 GGTGATTGAC GACATCACCG TTTTGATACA CGCGCAAAAA GAAGCCGCGT 1451 GGGGCGAAGT GGCGAAGCGG CTGGCACACG AAATCCGCAA TCCGCTCACG 1501 CCCATCCAGC TTTCCGCCGA ACGGCTGGCG TGGAAATTGG GCGGGAAGCT 1551 GGATGAGCAG GATGCGCAAA TCCTGACGCG TTCGACCGAC ACCATCGTCA 1601 AACAGGTGGC GGCATTGAAG GAAATGGTCG AAGCATTCCG CAATTATGCG 1651 CGTTCCCCTT CGCTCAAATT GGAAAATCAG GATTTGAACG CCTTAATCGG 1701 CGATGTGTTG GCATTGTATG AAGCCGGTCC GTGCCGGTTT GCGGCGGAGC 1751 TTGCCGGCGA ACCGCTGACG GTGGCGGCGG ATACGACCGC CATGCGGCAG 1801 GTGCTGCACA ATATTTTCAA AAATGCCGCC GAAGCGGCGG AAGAAGCCGA 1851 TGTGCCCGAA GTCAGGGTAA AATCGGAAAC AGGGCAGGAC GGTCGGATTG 1901 TCCTGACGGT TTGCGACAAC GGCAAAGGGT TCGGCAGGGA AATGCTGCAC 1951 AACGCCTTCG AGCCGTATGT AACGGACAAA CCGGCGGGAA CGGGATTGGG 2001 TCTGCCTGTG GTGAAAAAAA TCATTGAAGA ACACGGCGGC CGCATCAGCC 2051 TGAGCAATCA GGATGCGGGT GGCGCGTGTG TCAGAATCAT CTTGCCAAAA 2101 ACGGTAAAAA CTTATGCGTA G This corresponds to the amino acid sequence <SEQ ID 252; ORF64-l>: 1 MRRFLPIAAI CAWLLYGLT AATGSTSSLA DYFWWIVAFS AMLLLVLSAV 51 LARYVILLLK DRRDGVFGSQ IAKRLSGMFT LVAVLPGVFL FGVSAQFING 101 TINSWFGNDT HEALERSLNL SKSALNLAAD NALGNAVPVQ IDLIGAASLP 151 GDMGRVLEHY AGSGFAQLAL YNAASGKIEK SINPHKLDQP FPGKARWEKI 201 QRAGSVRDLE SIGGVLYAQG WLSAGTHNGR DYALFFRQPV PKGVAEDAVL 251 IEKARAKYAE LSYSKKGLQT FFLATLLIAS LLSIFLALVM ALYFARRFVE 301 PVLSLAEGAK AVAQGDFSQT RPVLRNDEFG RLTKLFNHMT EQLSIAKEAD 351 ERNRRREEAA RHYLECVLEG LTTGVWFDE QGCLKTFNKA AEQILGMPLT 401 PLWGSSRHGW HGVSAQQSLL AEVFAAIGAA AGTDKPVHVK YAAPDDAKIL 451 LGKATVLPED NGNGWMVID DITVLIHAQK EAAWGEVAKR LAHEIRNPLT 501 PIQLSAERLA WKLGGKLDEQ DAQILTRSTD TIVKQVAALK EMVEAFRNYA 551 RSPSLKLENQ DLNALIGDVL ALYEAGPCRF AAELAGEPLT VAADTTAMRQ 601 VLHNIFKNAA EAAEEADVPE VRVKSETGQD GRIVLTVCDN GKGFGREMLH 651 NAFEPYVTDK PAGTGLGLPV VKKIIEEHGG RISLSNQDAG GACVRIILPK 701 TVKTYA* Computer analysis of this amino acid sequence gave the following results: Homology with a predicted ORF from N.meninsitidis (strain A) ORF64 shows 92.6% identity over a 392aa overlap with an ORF (ORF64a) from strain A of N meningitidis: 10 20 30 40 50 60 orf 64 . pep MRRFLPIAAICAXXLXXGLTAATGSTSSLADYFWWIVAFSAMLLLVLSAVLARYVILLLK I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I orf 64a MRRFLPIAAICAVVLLYGLTAATGSTSSLADYFWWIVAFSAMLLLVLSAVLARYVILLLK 10 20 30 40 50 60 70 80 90 100 110 120 orf 64 . pep DRRDGVFGSXXAKXPXXXMFTLVAXLPGVFLFGFPAQFIΝGTIΝSWFGΝDTHEALERSLΝ I I I I I I II I II I I I I I I I I II II II II II II II II II I II I I II II II I I orf 64a DRRDGVFGSQIAKR-LSGMFTLVAVLPGVFLFGVSAQFIΝGTIΝSWFGΝDTHEALERSLΝ 70 80 90 100 110 130 140 150 160 170 180 orf 64 . pep LSKSALΝLAADΝALGΝAVPVQIDLIGAASLPGDMGRVLEHYAGSGFAQLALYΝXASGKIE I I I I I II II II I I II I I : I I II I I I II II I I II I II I I I I II I II II I I I I I I I I I I orf 64a LSKSALΝLAADΝALGΝAIPVQIDXIGAASLPXDMGRVLEHYAGSGFAQLALYΝAASGKIE 120 130 140 150 160 170 190 200 210 220 230 240 orf 64 . pep KSIΝPHKLDQPFPGKARWEKIQRAGSVRDLESIGGVLYAQGWLSAGTHXGRDYALFFRQP I I I II II II I II I II I I II II I: I I II I I I I II I II I I II II I II II II I I I I I II orf 64a KSIΝPHKLDQPFPGKARWEKIQQAGSVRDXESIGGVLYAXGWLSAXTHΝGRDYALFFRQP 180 190 200 210 220 230 250 260 270 280 290 300 orf 64 . pep VPKGVAEDAVLIEKARAKYAELSYSKKGLQTFFLATLLIASLLSIFLALVMALYFARRFV II I II I I II I II I I II I I I II I II II II I II I II II II I I I II I I I I I I I I I I I I I orf 64a VPKGVAEDAVLIEKARAXXXXLSYSKKGLQTFFLATLLIASLLSIFLALVMALYFARRFV 240 250 260 270 280 290 310 320 330 340 350 360 orf 64 . pep EPVLSLAEGAKAVAQGDFSQTRPVLRΝDEFGRLTXLFΝHMTEQLSIAKDADERΝRRREEA II I II I II II II II I I II I I II I I I I II II I II I I I II II II II II I : I I II II I II II orf 64a EPVLSLAEGAKAVAQGDFSQTRPVLRΝDEFGRLTKLFΝHMTEQLSIAKEADERΝRRREEA 300 310 320 330 340 350 370 380 390 orf 64 . pep ARHYLECVLEGLTTGVWFDEQGCLKTFΝKAAGT I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I orf 64a ARHYLECVLEGLTTGVWFDEQGCLKTFΝKAAEQILGMPLTPLWGSSRHGWHGVSAQQSL 360 370 380 390 400 410 orf 64 a LAEVFAAIGAAAGTDKPVHVKYAAPDDAKILLGKATVLPEDΝXΝGWMVIDDITVLIHAQ 420 430 440 450 460 470 The complete length ORF64a nucleotide sequence <SEQ ID 253> is: 1 ATGCGCCGTT TTCTACCGAT CGCAGCCATA TGCGCCGTCG TCCTGTTGTA 51 CGGACTGACG GCGGCAACCG GCAGCACCAG TTCGCTGGCG GATTATTTCT 101 GGTGGATTGT TGCGTTCAGC GCAATGCTGC TGCTGGTGTT GTCCGCCGTT 151 TTGGCACGTT ATGTCATATT GCTGTTGAAA GACAGGCGCG ACGGCGTATT 201 CGGTTCGCAG ATTGCCAAAC GCCTTTCCGG GATGTTTACG CTGGTTGCCG 251 TACTGCCCGG CGTGTTTCTG TTCGGCGTTT CCGCACAGTT TATCAACGGC 301 ACGATTAATT CGTGGTTCGG CAACGATACC CACGAGGCGC TTGAACGCAG 351 CCTCAATTTG AGCAAGTCCG CATTGAATCT GGCGGCAGAC AACGCCCTTG 401 GCAACGCCAT CCCCGTGCAG ATAGACΝTCA TCGGCGCGGC TTCCCTGCCC 451 ΝGGGATATGG GCAGGGTGCT GGAACATTAC GCCGGCAGCG GTTTTGCCCA 501 GCTTGCCCTG TACAATGCCG CAAGCGGCAA AATCGAAAAA AGCATCAACC 551 CGCACAAGCT CGATCAGCCG TTTCCAGGTA AGGCGCGTTG GGAAAAAATC 601 CAACAGGCGG GTTCGGTCAG GGATΝΝGGAA AGCATAGGCG GCGTATTGTA 651 CGCGCAΝGGC TGGCTGTCGG CAGΝΝACGCA CAACGGGCGC GATTACGCCT 701 TGTTTTTCCG TCAGCCGGTT CCCAAAGGCG TGGCAGAGGA TGCCGTCTTA 751 ATCGAAAAGG CAAGGGCGΝA AΝAΝΝΝTΝAG TTGAGTTACA GCAAAAAAGG 801 TTTGCAGACC TTTTTCCTΝG CAACCCTGCT GATTGCCTCΝ CTGCTGTCGA 851 TTTTTCTTGC ACTGGTCATG GCACTGTATT TCGCCCGCCG TTTCGTCGAA 901 CCCGTCCTAT CGCTTGCCGA GGGGGCGAAG GCGGTGGCGC AAGGCGATTT 951 CAGCCAGACG CGCCCCGTGT TGCGCAACGA CGAGTTCGGA CGCTTGACCA 1001 AGTTGTTCAA CCACATGACC GAGCAGCTTT CCATCGCCAA AGAAGCAGAC 1051 GAGCGCAACC GCCGGCGCGA GGAAGCCGCC AGACATTATC TCGAATGCGT 1101 GTTGGAGGGG CTGACCACGG GCGTGGTGGT GTTTGACGAA CAAGGCTGTC 1151 TGAAAACCTT CAACAAAGCG GCGGAACAGA TTTTGGGGAT GCCGCTTACC 1201 CCCCTGTGGG GCAGCAGCCG GCACGGTTGG CACGGCGTTT CGGCGCAGCA 1251 GTCCCTGCTT GCCGAAGTGT TTGCCGCCAT CGGCGCGGCG GCAGGTACGG 1301 ACAAACCGGT CCATGTGAAA TATGCCGCGC CGGACGATGC CAAAATCCTG 1351 CTGGGCAAGG CAACCGTCCT GCCCGAAGAC AACNGCAACG GCGTGGTAAT 1401 GGTGATTGAC GACATCACCG TTTTGATACA CGCGCAAAAA GAAGCCGCGT 1451 GGGGCGAAGT GGCAAAACGG CTGGCACACG AAATCCGCAA TCCGCTCACG 1501 CCCATCCAGC TTTCTGCCGA ACGGCTGGCG TGGAAATTGG GCGGGAAGCT 1551 GGACGAGCAN GACGCGCAAA TCCTGACACG TTCGACCGAC ACCATCATCA 1601 AACAAGTGGC GGCATTAAAA GAAATGGTCG AGGCATTCCG CAATTACNCG 1651 CGTTCCCCTT CGNCTCAATT GGAAAATCAG GATTTGAACG CCTTAATCGG 1701 CGATGTGTTG GCATTGTACG AAGCTGGTCC GTGCCGGTTT GCGGCGGAAC 1751 TTGCCGGCGA ACCGCTGATG ATGGCGGCGG ATACGACCGC CATGCGGCAG 1801 GTGCTGCACA ATATTTTCAA AAATGCCGCC GAAGCGGCGG AAGAAGCCGA 1851 TGTGCCCGAA GTCAGGGTAA AATCGGAAGC GGGGCAGGAC GGACGGATTG 1901 TCCTGACAGT TTGCGACAAC GGCAAGGGGT TCGGCAGGGA AATGCTGCAC 1951 AATGCCTTCG AGCCGTATGT AACGGACAAA CCGGCTGGAA CGGGATTGNG 2001 ACTGCCCGTG GTGAAAAAAA TCATTGAAGA ACACGGCGGC CNCATCAGCC 2051 TGAGCAATCA GGATGCGGGC GGCGCGTNTG TCAGAATCAT CTTGCCAAAA 2101 ACGGTAGAAA CTTATGCGTA G This encodes a protein having amino acid sequence <SEQ ID 254>: 1 MRRFLPIAAI CAWLLYGLT AATGSTSSLA DYFWWIVAFS AMLLLVLSAV 51 LARYVILLLK DRRDGVFGSQ IAKRLSGMFT LVAVLPGVFL FGVSAQFING 101 TINSWFGNDT HEALERSLNL SKSALNLAAD NALGNAIPVQ IDXIGAASLP 151 XDMGRVLEHY AGSGFAQLAL YNAASGKIEK SINPHKLDQP FPGKARWEKI 201 QQAGSVRDXE SIGGVLYAXG WLSAXTHNGR DYALFFRQPV PKGVAEDAVL 251 IEKARAXXXX LSYSKKGLQT FFLATLLIAS LLSIFLALVM ALYFARRFVE 301 PVLSLAEGAK AVAQGDFSQT RPVLRNDEFG RLTKLFNHMT EQLSIAKEAD 351 ERNRRREEAA RHYLECVLEG LTTGVWFDE QGCLKTFNKA AEQILGMPLT 401 PLWGSSRHGW HGVSAQQSLL AEVFAAIGAA AGTDKPVHVK YAAPDDAKIL 451 LGKATVLPED NXNGWMVID DITVLIHAQK EAAWGEVAKR LAHEIRNPLT 501 PIQLSAERLA WKLGGKLDEX DAQILTRSTD TIIKQVAALK EMVEAFRNYX 551 RSPSXQLENQ DLNALIGDVL ALYEAGPCRF AAELAGEPLM MAADTTAMRQ 601 VLHNIFKNAA EAAEEADVPE VRVKSEAGQD GRIVLTVCDN GKGFGREMLH 651 NAFEPYVTDK PAGTGLXLPV VKKIIEEHGG XISLSNQDAG GAXVRIILPK 701 TVETYA* ORF64a and ORF64-1 show 96.6% identity in 706 aa overlap: 10 20 30 40 50 60 orf64a . pep MRRFLPIAAICAWLLYGLTAATGSTSSLADYFWWIVAFSAMLLLVLSAVLARYVILLLK II I I I I I I I II I II I I II II I II I I II I II II II II I II I II I II II II II I II I I I I I I orf64-1 MRRFLPIAAICAWLLYGLTAATGSTSSLADYFWWIVAFSAMLLLVLSAVLARYVILLLK 10 20 30 40 50 60 70 80 90 100 110 120 orf64a . pep DRRDGVFGSQIAKRLSGMFTLVAVLPGVFLFGVSAQFINGTINSWFGNDTHEALERSLNL I I I I I I I II I I I II I I II I I I I II II I I I I II I II II II I II I I I II I I I I I II II I II I orf64-1 DRRDGVFGSQIAKRLSGMFTLVAVLPGVFLFGVSAQFINGTINSWFGNDTHEALERSLNL 70 80 90 100 110 120 130 140 150 160 170 180 orf64a . pep SKSALNLAADNALGNAIPVQIDXIGAASLPXDMGRVLEHYAGSGFAQLALYNAASGKIEK I I I II I II I II I I I MM II I I I II I I II I I II II I II II I II I I I I I I I I MM II I orf64-1 SKSALNLAADNALGNAVPVQIDLIGAASLPGDMGRVLEHYAGSGFAQLALYNAASGKIEK 130 140 150 160 170 180 190 200 210 220 230 240 orf64a. pep SINPHKLDQPFPGKARWEKIQQAGSVRDXESIGGVLYAXGWLSAXTHNGRDYALFFRQPV I II I I II II II II M I I II I M I II I II II I II II I I II I II I I II I II I I I I I I orf64-1 SINPHKLDQPFPGKARWEKIQRAGSVRDLESIGGVLYAQGWLSAGTHNGRDYALFFRQPV 190 200 210 220 230 240 250 260 270 280 290 300 orf64a . pep PKGVAEDAVLIEKARAXXXXLSYSKKGLQTFFLATLLIASLLSIFLALVMALYFARRFVE I II II II II II II II I I I II I II I II II I II I II II I II I I I II I I I II II I I I I I orf64-1 PKGVAEDAVLIEKARAKYAELSYSKKGLQTFFLATLLIASLLSIFLALVMALYFARRFVE 250 260 270 280 290 300 310 320 330 340 350 360 orf64a . pep PVLSLAEGAKAVAQGDFSQTRPVLRNDEFGRLTKLFNHMTEQLSIAKEADERNRRREEAA II I I II II I II II I II I I I I I I II I I II II II I I II II I I II I II II I I II I II II I II I orf64-1 PVLSLAEGAKAVAQGDFSQTRPVLRNDEFGRLTKLFNHMTEQLSIAKEADERNRRREEAA 310 320 330 340 350 360 370 380 390 400 410 420 orf64a.pep RHYLECVLEGLTTGVWFDEQGCLKTFNKAAEQILGMPLTPLWGSSRHGWHGVSAQQSLL I I II I I II II I I II I I I I II I I I II I II I II II I I II I I II I II I I II I I II I II I I I I I orf64-1 RHYLECVLEGLTTGVWFDEQGCLKTFNKAAEQILGMPLTPLWGSSRHGWHGVSAQQSLL 370 380 390 400 410 420 430 440 450 460 470 480 orf64a. pep AEVFAAIGAAAGTDKPVHVKYAAPDDAKILLGKATVLPEDNXNGWMVIDDITVLIHAQK I II II II II I II II II I I I I I I I II I I II II I II I II I I II I I I I I I I II I II I II II I orf64-1 AEVFAAIGAAAGTDKPVHVKYAAPDDAKILLGKATVLPEDNGNGWMVIDDITVLIHAQK 430 440 450 460 470 480 490 500 510 520 530 540 orf64a. pep EAAWGEVAKRLAHEIRNPLTPIQLSAERLAWKLGGKLDEXDAQILTRSTDTIIKQVAALK II I II II I I I I I I I II II II I I I II I II I II I II I II I I I I I I I I II I II I: I I I II II orf64-1 EAAWGEVAKRLAHEIRNPLTPIQLSAERLAWKLGGKLDEQDAQILTRSTDTIVKQVAALK 490 500 510 520 530 540 550 560 570 580 590 600 orf64a .pep EMVEAFRNYXRSPSXQLENQDLNALIGDVLALYEAGPCRFAAELAGEPLMMAADTTAMRQ I I II I I I II I I I I : II I I I I I II I I II II I II I II I II I II I I I I I I : I II I I I I II orf64-1 EMVEAFRNYARSPSLKLENQDLNALIGDVLALYEAGPCRFAAELAGEPLTVAADTTAMRQ 550 560 570 580 590 600 610 620 630 640 650 660 orf64a . pep VLHNIFKNAAEAAEEADVPEVRVKSEAGQDGRIVLTVCDNGKGFGREMLHNAFEPYVTDK I II I II II II I II II II I II I I I II I : II II II I I II I I II II I I I II I II I II I I I I I I orf64-1 VLHNIFKNAAEAAEEADVPEVRVKSETGQDGRIVLTVCDNGKGFGREMLHNAFEPYVTDK 610 620 630 640 650 660 670 680 690 700 orf64a.pep PAGTGLXLPWKKIIEEHGGXISLSNQDAGGAXVRIILPKTVETYAX II I II I I I I II II I I I II I II I I I I I I I I I I I II II I I I : I I I I orf64-l PAGTGLGLPWKKIIEEHGGRISLSNQDAGGACVRIILPKTVKTYAX 670 680 690 700 Homology with a predicted ORF from Nsonorrhoeae ORF 64 shows 86.6% identity over a 387aa overlap with a predicted ORF (ORF64.ng) from N gonorrhoeae: orf64.pep MRRFLPIAAICAXXLXXGLTAATGSTSSLADYFWWIVAFSAMLLLVLSAVLARYVILLLK 60 I I II I I I I I I II I II I I I II I I II I II I I II II: I II I I I I I I I I I I II II I I I I I orf64ng MRRFLPIAAICAWLLYGLTAATGSTSSLADYFWWIVSFSAMLLLVLSAVLARYVILLLK 60 orf64.pep DRRDGVFGSXXAKXPXXXMFTLVAXLPGVFLFGFPAQFIΝGTIΝSWFGΝDTHEALERSLΝ 120 I II: II II I II I II II I II M I II M II I I I I I II I I II I II I II I I I I I I orf64ng DRRΝGVFGSQIAKR-LSGMFTLVAVLPGLFLFGISAQFIΝGTIΝSWFGΝDTHEALERSLΝ 119 orf64.pep LSKSALΝLAADΝALGΝAVPVQIDLIGAASLPGDMGRVLEHYAGSGFAQLALYΝXASGKIE 180 I I II II : II II II :: II I II I I I II I : II I MM I I II I II II II I II I II II I II I orf64ng LSKSALDLAADΝAVSΝAVPVQIDLIGTASLSGΝMGSVLEHYAGSGFAQLALYΝAASGKIE 179 orf64.pep KSIΝPHKLDQPFPGKARWEKIQRAGSVRDLESIGGVLYAQGWLSAGTHXGRDYALFFRQP 240 j I I I IM: I IM I I : I I : I I :: I II I : II II I I II I I II I I I I I I I I I II II I I II I orf64ng KSIΝPHQFDQPLPDKEHWEQIQQTGSVRSLESIGGVLYAQGWLSAGTHΝGRDYALFFRQP 239 orf 64 . pep VPKGVAEDAVLIEKARAKYAELSYSKKGLQTFFLATLLIASLLSIFLALVMALYFARRFV 300 M :: I I : II II I I II I II II II I I II I II I I II I : I II I II I II II I I I I II I I I I I I I I orf 64ng IPENVAQDAVLIEKARAKYAELSYSKKGLQTFFLVTLLIASLLSIFLALVMALYFARRFV 299 orf 64 . pep EPVLSLAEGAKAVAQGDFSQTRPVLRNDEFGRLTXLFNHMTEQLSIAKDADERNRRREEA 360 I I : I II I II I II II I II I I I I I II II II I II I II II II II I II II II : I II I I I I I I I I orf 64ng EPILSLAEGAKAVAQGDFSQTRPVLRNDEFGRLTKLFNHMTEQLSIAKEADERNRRREEA 359 orf64. pep ARHYLECVLEGLTTGVWFDEQGCLKTFNKAAGT 394 I I I I I II II : II I II II I : I : 1 orf64ng ARHYLECVLDGLTTGVWSYPLSCCRTAVFSTCHSSPLSYF 400 An ORF64ng nucleotide sequence <SEQ LD 255> was predicted to encode a protein having amino acid sequence <SEQ LD 256>: 1 MRRFLPIAAI CAWLLYGLT AATGSTSSLA DYFWWIVSFS AMLLLVLSAV 51 LARYVILLLK DRRNGVFGSQ IAKRLSGMFT LVAVLPGLFL FGISAQFING 101 TINSWFGNDT HEALERSLNL SKSALDLAAD NAVSNAVPVQ IDLIGTASLS 151 GNMGSVLEHY AGSGFAQLAL YNAASGKIEK SINPHQFDQP LPDKEHWEQI 201 QQTGSVRSLE SIGGVLYAQG WLSAGTHNGR DYALFFRQPI PENVAQDAVL 251 IEKARAKYAE LSYSKKGLQT FFLVTLLIAS LLSIFLALVM ALYFARRFVE 301 PILSLAEGAK AVAQGDFSQT RPVLRNDEFG RLTKLFNHMT EQLSIAKEAD 351 ERNRRREEAA RHYLECVLDG LTTGVWSYP LSCCRTAVFS TCHSSPLSYF* Further work revealed the complete gonococcal DNA sequence <SEQ LD 257>: 1 ATGCGCCGCT TCCTACCGAT CGCAGCCATA TGCGCCGTCG TCCTGCTGTA 51 CGGATTGACG GCGGCGACCG GCAGCACCAG TTCGCTGGCG GATTATTTCT 101 GGTGGATAGT CTCGTTCAGC GCAATGCTGC TGCTGGTGTT GTCCGCCGTT 151 TTGGCACGTT ATGTCATATT GCTGTTGAAA GACAGGCGCA ACGGCGTGTT 201 CGGTTCGCAG ATTGCCAAAC GCCTTTCCGG GATGTTCACG CTGGTCGCCG 251 TACTGCCCGG CTTGTTCCTG TTCGGCATTT CCGCGCAGTT TATCAACGGC 301 ACGATTAATT CGTGGTTCGG CAACGACACC CACGAAGCCC TCGAACGCAG 351 CCTTAATTTG AGCAAGTCCG CACTGGATTT GGCGGCAGAC AATGCCGTCA 401 GCAACGCCGT TCCCGTACAG ATAGACCTCA TCGGCACCGC CTCCCTGTCG 451 GGCAATATGG GCAGTGTGCT GGAACACTAC GCCGGCAGCG GTTTTGCCCA 501 GCTTGCCCTG TACAATGCCG CAAGCGGGAA AATCGAAAAA AGCATCAATC 551 CGCACCAATT CGACCAGCCG CTTCCCGACA AAGAACATTG GGAACAGATT 601 CAGCAGACCG GTTCGGTTCG GAGTTTGGAA AGCATAGGCG GCGTATTGTA 651 CGCGCAGGGA TGGTTGTCGG CAGGTACGCA CAACGGGCGC GATTACGCGC 701 TGTTCTTCCG CCAGCCGATT CCCGAAAATG TGGCACAGGA TGCCGTTCTG 751 ATTGAAAAGG CGCGGGCGAA ATATGCCGAA TTGAGTTACA GCAAAAAAGG 801 TTTGCAGACC TTTTTTCTGG TAACCCTGCT GATTGCCTCG CTGCTGTCGA 851 TTTTTCTTGC GCTGGTAATG GCACTGTATT TTGCCCGCCG TTTCGTCGAA 901 CCCATTCTGT CGCTTGCCGA GGGCGCAAAG GCGGTGGCGC AGGGTGATTT 951 CAGCCAGACG CGCCCCGTAT TGCGCAACGA CGAGTTCGGA CGTTTGACCA 1001 AGCTGTTCAA CCATATGACC GAGCAGCTTT CCATCGCCAA AGAAGCAGAC 1051 GAACGCAACC GCCGGCGCGA GGAAGCCGCC CGTCACTACC TCGAGTGCGT 1101 GTTGGATGGG TTGACTACCG GTGTGGTGGT GTTTGACGAA AAAGGCCGTT 1151 TGAAAACCTT CAACAAGGCG GCGGAACAGA TTTTGGGGAT GCCGCTCGCC 1201 CCCCTGTGGG GCAGCAGCCG GCACGGTTGG CACGGCGTTT CGGCGCAGCA 1251 GTCCCTGCTT GCCGAAGTGT TtgccgccAT CGGTGCGGCG GCAGGTACGG 1301 ACAAACCGGT CCAGGTGGAA TATGCCGCGC CGGACGATGC CAAAATCCTG 1351 CTGGGCAAGG CGACGGTATT GCCCGAAGAC AACGGCAACG GCGTGGTGAT 1401 GGTGATTGAC GACATCACCG TGCTGATACG CGCGCAAAAA GAAGCCGCGT 1451 GGGGTGAAGT GGCGAAGCGG CTGGCACACG AAATCCGCAA TCCGCTCACG 1501 CCCATCCAGC TTTCCGCCGA ACGGCTGGCG TGGAAATTGG GCGGGAAGCT 1551 GGACGATCAG GACGCGCAAA TCCTGACGCG TtcgACCGAC ACCATCATCA 1601 AACAGgtggc gGCGTTAAAA GAAATGGTCG AGGCATTCCG CAATTACGCG 1651 CGCGCCCCTT CGCTCAAACT GGAAAATCAG GATTTGAACG CCTTAATCGG 1701 CGATGTTTTG GCCCTGTACG AAGCCGGCCC GTGCCGGTTT GAGGCGGAAC 1751 TTGCCGGCGA ACCGCTGATG ATGGCGGCGG ATACGACCGC CATGCGGCAG 1801 GTGCTGCACA ATATTTTCAA AAATGCCGCC GAAGCGGCGG AAGAAGCCGA 1851 TATGCCCGAA GTCAGGGTAA AATCGGAAAC GGGGCAGGAC GGACGGATTG 1901 TCCTGACGGT TTGCGACAAC GGCAAGGGAT TCGGCAAGGA AATGCTGCAC 1951 AATGCTTTCG AGCCGTATGT GACGGATAAG CCGGCGGGAA CGGGACTGGG 2001 TCTGCCTGTA GTGAAAAAAA TCATTGGAGA ACACGGCGGC CGCATCAGCC 2051 TGAGCAATCA GGATGCGGGT GGGGCGTGTG TCAGAATCAT CTTGCCAAAA 2101 ACGGTAGAAA CTTATGCGTA G This corresponds to the amino acid sequence <SEQ ID 258; ORF64ng-l > • 1 MRRFLPIAAI CAWLLYGLT AATGSTSSLA DYFWWIVSFS AMLLLVLSAV 51 LARYVILLLK DRRNGVFGSQ IAKRLSGMFT LVAVLPGLFL FGISAQFING 101 TINSWFGNDT HEALERSLNL SKSALDLAAD NAVSNAVPVQ IDLIGTASLS 151 GNMGSVLEHY AGSGFAQLAL YNAASGKIEK SINPHQFDQP LPDKEHWEQI 201 QQTGSVRSLE SIGGVLYAQG WLSAGTHNGR DYALFFRQPI PENVAQDAVL 251 IEKARAKYAE LSYSKKGLQT FFLVTLLIAS LLSIFLALVM ALYFARRFVE 301 PILSLAEGAK AVAQGDFSQT RPVLRNDEFG RLTKLFNHMT EQLSIAKEAD 351 ERNRRREEAA RHYLECVLDG LTTGVWFDE KGRLKTFNKA AEQILGMPLA 401 PLWGSSRHGW HGVSAQQSLL AEVFAAIGAA AGTDKPVQVE YAAPDDAKIL 451 LGKATVLPED NGNGWMVID DITVLIRAQK EAAWGEVAKR LAHEIRNPLT 501 PIQLSAERLA WKLGGKLDDQ DAQILTRSTD TIIKQVAALK EMVEAFRNYA 551 RAPSLKLENQ DLNALIGDVL ALYEAGPCRF EAELAGEPLM MAADTTAMRQ 601 VLHNIFKNAA EAAEEADMPE VRVKSETGQD GRIVLTVCDN GKGFGKEMLH 651 NAFEPYVTDK PAGTGLGLPV VKKIIGEHGG RISLSNQDAG GACVRIILPK 701 TVETYA* ORF64ng-l and ORF64-1 show 93.8% identity in 706 aa overlap: 10 20 30 40 50 60 orf64ng-l.pep MRRFLPIAAICAWLLYGLTAATGSTSSLADYFWWIVSFSAMLLLVLSAVLARYVILLLK II I II II I II I II II II I I II I I I II I II I II I I I I I : II II I I I I I I I II II I I I I II I orf64-l MRRFLPIAAICAWLLYGLTAATGSTSSLADYFWWIVAFSAMLLLVLSAVLARYVILLLK 10 20 30 40 50 60 70 80 90 100 110 120 orf64ng-l.pep DRRNGVFGSQIAKRLSGMFTLVAVLPGLFLFGISAQFINGTINSWFGNDTHEALERSLNL I I I: II I II I II II I II I II I I I I II I: I I II: I I I I I II II I I I I I I I I I II I I I I II I orf64-l DRRDGVFGSQIAKRLSGMFTLVAVLPGVFLFGVSAQFINGTINSWFGNDTHEALERSLNL 70 80 90 100 110 120 130 140 150 160 170 180 orf64ng-l .pep SKSALDLAADNAVSNAVPVQIDLIGTASLSGNMGSVLEHYAGSGFAQLALYNAASGKIEK II I II: I I II I M: II II I I II I II: II I I: I I II II I I II I II I I I I II I I I I I II I orf64-l SKSALNLAADNALGNAVPVQIDLIGAASLPGDMGRVLEHYAGSGFAQLALYNAASGKIEK 130 140 150 160 170 180 190 200 210 220 230 240 orf64ng-l .pep SINPHQFDQPLPDKEHWEQIQQTGSVRSLESIGGVLYAQGWLSAGTHNGRDYALFFRQPI II I IMMI I : I I : I I: IM: I II I: I I I I I I II II I I I I II I II I I II I I I I II I I : orf64-l SINPHKLDQPFPGKARWEKIQRAGSVRDLESIGGVLYAQGWLSAGTHNGRDYALFFRQPV 190 200 210 220 230 240 250 260 270 280 290 300 orf64ng-l.pep PENVAQDAVLIEKARAKYAELΞYSKKGLQTFFLVTLLIASLLSIFLALVMALYFARRFVE M: IM II I I II II I II I I II I II I II I II I II M I I I I II I II II I I I I II I I I I I II I orf64-l PKGVAEDAVLIEKARAKYAELSYSKKGLQTFFLATLLIASLLSIFLALVMALYFARRFVE 250 260 270 280 290 300 310 320 330 340 350 360 orf64ng-l .pep PILSLAEGAKAVAQGDFSQTRPVLRNDEFGRLTKLFNHMTEQLSIAKEADERNRRREEAA I: II II I II I M I II I I II I II I II I II I II I I I II I II I I I I II I I M II I II II I I II orf64-l PVLSLAEGAKAVAQGDFSQTRPVLRNDEFGRLTKLFNHMTEQLSIAKEADERNRRREEAA 310 320 330 340 350 360 370 380 390 400 410 420 orf64ng-l .pep RHYLECVLDGLTTGVWFDEKGRLKTFNKAAEQILGMPLAPLWGSSRHGWHGVSAQQSLL II I I II II : I I I I II I I I I I: I I I II I II I II II II I I I I I II I I I I I II II I I I II orf64-l RHYLECVLEGLTTGWVFDEQGCLKTFNKAAEQILGMPLTPLWGSSRHGWHGVSAQQSLL 370 380 390 400 410 420 430 440 450 460 470 480 orf64ng-l .pep AEVFAAIGAAAGTDKPVQVEYAAPDDAKILLGKATVLPEDNGNGWMVIDDITVLIRAQK I I I I I I I I I II I I I I 11:1:11 II I I II I II I I I I I I I II II I II II I I I II I I I 1:11 I orf64-l AEVFAAIGAAAGTDKPVHVKYAAPDDAKILLGKATVLPEDNGNGWMVIDDITVLIHAQK 430 440 450 460 470 480 490 500 510 520 530 540 orf64ng-l.pep EAAWGEVAKRLAHEIRNPLTPIQLSAERLAWKLGGKLDDQDAQILTRSTDTIIKQVAALK III I M I II I II I I I II I II I I II I II I II I II I II 11:1 I I I II II II I ||:| I I I II I orf64-l EAAWGEVAKRLAHEIRNPLTPIQLSAERLAWKLGGKLDEQDAQILTRSTDTIVKQVAALK 490 500 510 520 530 540 550 560 570 580 590 600 orf 64ng-l . pep EMVEAFRNYARAPSLKLENQDLNALIGDVLALYEAGPCRFEAELAGEPLMMAADTTAMRQ II II I I I I I I I : I I I I I II I II II I II II II II I II II II I II II I I I : I I I I I I II I orf64-l EMVEAFRNYARSPSLKLENQDLNALIGDVLALYEAGPCRFAAELAGEPLTVAADTTAMRQ 550 560 570 580 590 600 610 620 630 640 650 660 orf64ng-l.pep VLHNIFKNAAEAAEEADMPEVRVKSETGQDGRIVLTVCDNGKGFGKEMLHNAFEPYVTDK II II II I I I II II I II I: II I II II I I I I II I II I II II II I I II: II II I I I I I I II I I orf64-l VLHNIFKNAAEAAEEADVPEVRVKSETGQDGRIVLTVCDNGKGFGREMLHNAFEPYVTDK 610 620 630 640 650 660 670 680 690 700 orf64ng-l .pep PAGTGLGLPWKKIIGEHGGRISLSNQDAGGACVRIILPKTVETYAX II II II II I II II II II II II II II II II II I I II I I II II: I II I orf64-l PAGTGLGLPWKKIIEEHGGRISLSNQDAGGACVRIILPKTVKTYAX 670 680 690 700 Furthermore, ORF64ng-l shows significant homology to a protein from A. caulinodans: sp|Q04850|NTRY_AZOCA NITROGEN REGULATION PROTEIN NTRY >gi | 77479 |pir || S18624 ntrY protein - Azorhizobium caulinodans >gi 138737 (X63841) NtrY gene product [Azorhizobium caulinodans] Length = 771 Score = 218 bits (550), Expect = 7e-56 Identities = 195/720 (27%), Positives •= 320/720 (44%), Gaps = 58/720 (8%) Query: 7 IAAICAWLLYGLTAATGSTSSLADYFWWIXXXXXXXXXXXXXXXXRYVILLLKDRRNGV 66 I+A+ ++L GLT + + + R + + K R G Sbjct: 35 ISALATFLILMGLTPWPTHQWIS VLLVNAAAVLILSAMVGREIWRIAKARARGR 90 Query: 67 FGSQIAKRLSGMFTLVAVLPGLFLFGISAQFINGTINSWFGNDTHEALERSLNLSKSALD 126 +++ R+ G+F +V+V+P + + +++ ++ ++ WF T E + S++++++ + Sbjct: 91 AAARLHIRIVGLFAWSWPAILVAWASLTLDRGLDRWFSMRTQEIVASSVSVAQTYVR 150 Query: 127 LAADNAVSNAVPVQIDLIGTASLSGNMGSVLEHYAG—SGFAQLALYNAASGKIEKSINP 184 A N + + + DL S+ Y G S F Q+ AA + ++ Sbjct: 151 EHALNIRGDILAMSADLTRLKSV YEGDRSRFNQILTAQAALRNLPGAMLI 200 Query: 185 HQFDQPLPDKEHWEQIQQTGSVRSLESIGGVLYAQGWLSAGTHNGRDYA 233 + D + ++ + 1 + V + +IG Q + N DY Sbjct: 201 RR-DLSWERAN-VNIGREFIVPANLAIGDATPDQPVIYLP—NDADYVAAWPLKDYDD 256 Query: 234 —LFFRQPIPENVAQDAVLIEKARAKYAELSYSKKGLQTFFLVTXXXXXXXXXXXXXVMA 291 L+ + I V ++ A Y L + G+Q F + + Sbjct: 257 LYLYVARLIDPRVIGYLKTTQETLADYRSLEERRFGVQVAFALMYAVITLIVLLSAVWLG 316 Query: 292 LYFARRFVEPILSLAEGAKAVAQGDFSQTRPVLRND-EFGRLTKLFNHMTEQLSIXXXXX 350 L F++ V PI L A VA+G+ P+ R + + L + FN MT +L Sbjct: 317 LNFSKWLVAPIRRLMSAADHVAEGNLDVRVPIYRAEGDLASLAETFNKMTHELRSQREAI 376 Query: 351 XXXXXXXXXXXHYLECVLDGLTTGVWFDEKGRLKTFNKAAEQILGMPLAPLWGSSRHGW 410 + E VL G+ GV+ D + R+ N++AE++LG L+ + RH Sbjct: 377 LTARDQIDSRRRFTEAVLSGVGAGVIGLDSQERITILNRSAERLLG—LSEVEALHRHLA 434 Query: 411 HGVSAQQSLLAEVFXXXXXXXXTDKPVQVEYAAPDDAKILLGKATVLPEDNG NGWM 467 V LL E + VQ D + + V E + +G V+ Sbjct: 435 EWPETAGLLEEA EHARQRSVQGNITLTRDGRERVFAVRVTTEQSPEAEHGWW 488 Query: 468 VIDDITVLIRAQKEAAWGEVAKRLAHEIRNPLTPIQLSAERLAWKLGGKLDDQDAQILTR 527 +DDIT LI AQ+ +AW +VA+R+AHEI+NPLTPIQLSAERL K G + QD +1 + Sbjct: 489 TLDDITELISAQRTSAWADVARRIAHEIKNPLTPIQLSAERLKRKFGRHV-TQDREIFDQ 547 Query: 528 STDTIIKQVAALKEMVEAFRNYARAPSLKLENQDLNALIGDVLALYEAGPCRFEAELAGE 587 TDTII+QV + MV+ F ++AR P +++QD++ +1 + L G + Sbjct: 548 CTDTIIRQVGDIGRMVDEFSSFARMPKPWDSQDMSEIIRQTVFLMRVGHPEWFDSEVP 607 Query: 588 PLMMAA-DTTAMRQVLHNIFKNXXXXXXXXDMPEVRVK SETGQDGRIVLTVCD 639 P M A D + Q L NI KN P+VR + + G+D +V+ + D Sbjct: 608 PAMPARFDRRLVSQALTNILKNAAEAIEAVP-PDVRGQGRIRVSANRVGED—LVIDIID 664 Query : 640 NGKGFGKEMLHNAFEPYVTDKPAGTGLGLPWKKIIGEHGGRISLSNQDAG-GACVRIIL 698 NG G +E + EPYVT + GTGLGL +V KI+ EHGG I L++ G GA +R+ L Sbj ct : 665 NGTGLPQESRNRLLEPYVTTREKGTGLGLAIVGKIMEEHGGGIELNDAPEGRGAWIRLTL 724 Based on this analysis, including the presence of a putative leader sequence (double-underlined) and several putative transmembrane domains (single-underlined) in the gonococcal protein, it is predicted that the proteins from N. meningitidis and N.gonorrhoeae, and their epitopes, could be useful antigens for vaccines or diagnostics, or for raising antibodies. Example 31 The following partial DNA sequence was identified in N meningitidis <SEQ ID 259>: 1 ATGTACGCAT TTACCGCCGC ACAGCAACAG AAGGCACTCT TCCGGCTGGT 51 GCTTTTTCAT ATCCTCATCA TCGCCGCCAG CAACTATCTG GTGCAGTTCC 101 CTTTCCAAAT TTTCGGCATC CACACCACTT GGGGCGCATT TTCCTTTCCC 151 TTCATCTTCC TTGCCACCGA CCTGACCGTC CGCATTTTCG GTTCTCACTT 201 GGCACGGCGG ATTATCTTTT GGGTGATGTT CCCCGCCCTT TTGCTTTCCT 251 ACGTCTTTTC CGTTTTGTTC CACAACGGCA GTTGGACAGG CTTGGGCGCG 301 CTGTCCGAAT TCAACACCTT TGTCGGACGC ATCGCCTTAG CCAGCTTTGC 351 CGCCTACGCG ATCGGACAAA TCCTTGATAT TTTTGTATTC AACAAATTAC 401 GCCGTCTGAA AGCGTGGTGG ATTGCACCGA ACGCATCAAC CGTCATCGGG 451 CACGCGTTGG ATACG... This corresponds to the amino acid sequence <SEQ ID 260; ORF66>: 1 MYAFTAAQQQ KALFRLVLFH ILIIAASΝYL VQFPFQIFGI HTTWGAFSFP 51 FIFLATDLTV RIFGSHLARR IIFWVMFPAL LLSYVFSVLF HΝGSWTGLGA 101 LSEFΝTFVGR IALASFAAYA IGQILDIFVF ΝKLRRLKAWW IAPΝAΞTVIG 151 HALDT... Further work revealed the complete nucleotide sequence <SEQ ID 261>: 1 ATGTACGCAT TTACCGCCGC ACAGCAACAG AAGGCACTCT TCCGGCTGGT 51 GCTTTTTCAT ATCCTCATCA TCGCCGCCAG CAACTATCTG GTGCAGTTCC 101 CTTTCCAAAT TTTCGGCATC CACACCACTT GGGGCGCATT TTCCTTTCCC 151 TTCATCTTCC TTGCCACCGA CCTGACCGTC CGCATTTTCG GTTCTCACTT 201 GGCACGGCGG ATTATCTTTT GGGTGATGTT CCCCGCCCTT TTGCTTTCCT 251 ACGTCTTTTC CGTTTTGTTC CACAACGGCA GTTGGACAGG CTTGGGCGCG 301 CTGTCCGAAT TCAACACCTT TGTCGGACGC ATCGCCTTAG CCAGCTTTGC 351 CGCCTACGCG ATCGGACAAA TCCTTGATAT TTTTGTATTC AACAAATTAC 401 GCCGTCTGAA AGCGTGGTGG ATTGCACCGA CCGCATCAAC CGTCATCGGC 451 AACGCCTTGG ATACGCTGGT ATTTTTCGCC GTTGCCTTCT ACGCAAGCAG 501 CGATGGATTT ATGGCGGCAA ACTGGCAGGG CATCGCTTTT GTCGATTACC 551 TGTTCAAACT TACCGTCTGC ACCCTCTTCT TCCTGCCCGC CTACGGCGTG 601 ATACTGAATC TGCTGACGAA AAAACTGACA ACCCTGCAAA CCAAACAGGC 651 GCAAGACCGC CCCGCGCCCT CGCTGCAAAA TCCGTAA This corresponds to the amino acid sequence <SEQ LD 262; ORF66-l>: 1 MYAFTAAQQQ KALFRLVLFH ILIIAASΝYL VQFPFQIFGI HTTWGAFSFP 51 FIFLATDLTV RIFGSHLARR IIFWVMFPAL LLSYVFSVLF HΝGSWTGLGA 101 LSEFΝTFVGR IALASFAAYA IGQILDIFVF ΝKLRRLKAWW IAPTASTVIG 151 ΝALDTLVFFA VAFYASSDGF MAAΝWQGIAF VDYLFKLTVC TLFFLPAYGV 201 ILΝLLTKKLT TLQTKQAQDR PAPSLQΝP* Computer analysis of this amino acid sequence gave the following results: Homology with the hypothetical protein o221 of E. coli (accession number P376191 ORF66 and o221 protein show 67% aa identity in 155aa overlap: orf66 1 MYAFTAAQQQKALFRLVLFHILIIAASNYLVQFPFQIFGIHTTWGAFSFPFIFLATDLTV 60 M F+ Q+ KALF L LFH+L+I +SNYLVQ P I G HTTWGAFSFPFIFLATDLTV o221 1 MNVFSQTQRYKALFWLSLFHLLVITSSNYLVQLPVSILGFHTTWGAFSFPFIFLATDLTV 60 orf66 61 RIFGSHLARRIIFWVMFPALLLSYVFSVLFHNGSWTGLGALSEFNTFVGRIALASFAAYA 120 RIFG+ LARRIIF VM PALL+SYV S LF+ GSW G GAL+ FN FV RIA ASF AYA o221 61 RIFGAPLARRIIFAVMIPALLISYVISSLFYMGSWQGFGALAHFNLFVARIATASFMAYA 120 orf66 121 IGQILDIFVFNKLRRLKAWWIAPNASTVIGHALDT 155 +GQILD+ VFN+LR+ + WW+AP AST+ G+ DT o221 121 LGQILDVHVFNRLRQSRRWWLAPTASTLFGNVSDT 155 Homology with a predicted ORF from N.meninsitidis (strain A) ORF66 shows 96.1% identity over a 155aa overlap with an ORF (ORF66a) from strain A of N meningitidis: 10 20 30 40 50 60 orf 66 . pep MYAFTAAQQQKALFRLVLFHILIIAASΝYLVQFPFQIFGIHTTWGAFSFPFIFLATDLTV I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I orf 66a MYAFTAAQQQKALFWLVLFHILIIAASΝYLVQFPFQISGIHTTWGAFSFPFIFLATDLTV 10 20 30 40 50 60 70 80 90 100 110 120 orf 66 . pep RIFGSHLARRIIFWVMFPALLLSYVFSVLFHΝGSWTGLGALSEFΝTFVGRIALASFAAYA I I II I II II II I II II I II I I I I II I I I I I II I I I I I II II I II I II I I I I I I II II I I I orf 66a RIFGSHLARRIIFWVMFPALLLSYVFSVLFHΝGSWTGLGALSEFΝTFVGRIALASFAAYA 70 30 90 100 110 120 130 140 150 orf 66 . pep IGQILDIFVFΝKLRRLKAWWIAPΝASTVIGHALDT : II I II II II I II II II I II : I I M I I II I: I II I orf 66a LGQILDIFVFΝKLRRLKAWWVAPTASTVIGΝALDTLVFFAVAFYASSDGFMAAΝWQGIAF 130 140 150 160 170 180 orf 66a VDYLFKLTVCGLFFLPAYGVILΝLLTKKLTTLQTKQAQDRPAPSLQΝPX 190 200 210 220 The complete length ORF66a nucleotide sequence <SEQ ID 263> is: 1 ATGTACGCAT TTACCGCCGC ACAGCAACAG AAGGCACTCT TCTGGCTGGT 51 GCTTTTTCAT ATCCTCATCA TCGCCGCCAG CAACTATCTG GTGCAGTTCC 101 CCTTCCAAAT TTCCGGCATC CACACCACTT GGGGCGCGTT TTCCTTTCCC 151 TTCATCTTCC TCGCCACCGA CCTGACCGTC CGCATTTTCG GTTCGCACTT 201 GGCACGGCGG ATTATCTTTT GGGTCATGTT CCCCGCCCTT TTGCTTTCCT 251 ACGTCTTTTC CGTTTTGTTC CACAACGGCA GTTGGACGGG CTTGGGCGCG 301 CTGTCCGAAT TCAACACCTT TGTCGGACGC ATCGCGCTGG CAAGTTTTGC 351 CGCCTACGCG CTCGGACAAA TCCTTGATAT TTTTGTGTTC AACAAATTAC 401 GCCGTCTGAA AGCGTGGTGG GTTGCCCCGA CTGCATCAAC CGTCATCGGC 451 AACGCCTTAG ATACGTTGGT ATTTTTCGCC GTTGCCTTCT ACGCAAGCAG 501 CGATGGATTT ATGGCGGCAA ACTGGCAGGG CATCGCTTTT GTCGATTACC 551 TGTTCAAACT CACCGTCTGC GGTCTGTTTT TCCTGCCCGC CTACGGCGTG 601 ATTCTGAATC TGCTGACGAA AAAACTGACG ACCCTGCAAA CCAAACAGGC 651 GCAAGACCGC CCCGCGCCCT CGCTGCAAAA TCCGTAA This encodes a protein having amino acid sequence <SEQ ID 264>: 1 MYAFTAAQQQ KALFWLVLFH ILIIAASΝYL VQFPFQISGI HTTWGAFSFP 51 FIFLATDLTV RIFGSHLARR IIFWVMFPAL LLSYVFSVLF HΝGSWTGLGA 101 LSEFΝTFVGR IALASFAAYA LGQILDIFVF ΝKLRRLKAWW VAPTASTVIG 151 ΝALDTLVFFA VAFYASSDGF MAAΝWQGIAF VDYLFKLTVC GLFFLPAYGV 201 ILΝLLTKKLT TLQTKQAQDR PAPSLQΝP* ORF66a and ORF66-1 show 97.8% identity in 228 aa overlap: 10 20 30 40 50 60 orf66a . pep MYAFTAAQQQKALFWLVLFHILIIAASΝYLVQFPFQISGIHTTWGAFSFPFIFLATDLTV II I II I II I II I II II I I I I I I I II II II I I I I II I I I I I II II I II I II II I I I I I I orf66-1 MYAFTAAQQQKALFRLVLFHILIIAASΝYLVQFPFQIFGIHTTWGAFSFPFIFLATDLTV 10 20 30 40 50 60 70 80 90 100 110 120 orf66a . pep RIFGSHLARRIIFWVMFPALLLSYVFSVLFHNGSWTGLGALSEFNTFVGRIALASFAAYA I I II I I II II I I II II I II II II I II II I I II II I II I I II I I I I I I I I I I I I I I I I I II orf66-1 RIFGSHLARRIIFWVMFPALLLSYVFSVLFHNGSWTGLGALSEFNTFVGRIALASFAAYA 70 80 90 100 110 120 130 140 150 160 170 180 orf66a. pep LGQILDIFVFNKLRRLKAWWVAPTASTVIGNALDTLVFFAVAFYASSDGFMAANWQGIAF : I I II II I II II I I I II I IM II I II II II II II I II I I II I I I I I II I I I II I I I I I I I orf66-1 IGQILDIFVFNKLRRLKAWWIAPTASTVIGNALDTLVFFAVAFYASSDGFMAANWQGIAF 130 140 150 160 170 180 190 200 210 220 229 orf66a . pep VDYLFKLTVCGLFFLPAYGVILNLLTKKLTTLQTKQAQDRPAPSLQNPX I I II I I I II I I I I I I I I I I I II I II II II I I II I II II II II II I II I orf66-1 VDYLFKLTVCTLFFLPAYGVILNLLTKKLTTLQTKQAQDRPAPSLQNPX 190 200 210 220 Homology with a predicted ORF from N.sonorrhoeae ORF66shows 94.2% identity over a 155aa overlap with a predicted ORF (ORF66.ng) from N gonorrhoeae: orf66. pep MYAFTAAQQQKALFRLVLFHILIIAASΝYLVQFPFQIFGIHTTWGAFSFPFIFLATDLTV 60 I II : I I I II II I I I I II I I II I II I II I I II II I I : I II II I II I I II I I II II I II II I orf66ng MYALTAAQQQKALFRLVLFHILIIAASΝYLVQFPFRIFGIHTTWGAFSFPFIFLATDLTV 60 orf66. pep RIFGSHLARRIIFWVMFPALLLSYVFSVLFHΝGSWTGLGALSEFΝTFVGRIALASFAAYA 120 II I I I I II I II I I I II I I II I II I II I I II II I II I M I I : I II II I I II I I I I I I I I orf66ng RIFGSHLARRIIFWVMFPALSLSYVFSVLFHΝGSWTGLGAPSQFΝTFVGRIALASFAAYA 120 orf66. pep IGQILDIFVFΝKLRRLKAWWIAPΝASTVIGHALDT 155 : I I II II I II: I II I I I I I I I II II II I M I I II orf66ng LGQILDIFVFDKLRRLKAWWIAPAASTVIGΝALDTLVFFAVAFYASSDEFMAAΝWQGIAF 180 The complete length ORF66ng nucleotide sequence <SEQ ID 265> is: 1 ATGTACGCAT TGACCGCCGC ACAGCAACAG AAGGCACTCT TCCGGCTGGT 51 GCTTTTCCAT ATCCTCATCA TCGCCGCCAG CAACTATCTG GTGCAGTTCC 101 CCTTCCGGAT TTTCGGCATC CACACCACTT GGGGCGCGTT TTCCTTTCCC 151 TTCATCTTCC TCGCCACCGA CCTGACCGTC CGCATTTTCG GTTCGCACTT 201 GGCGCGGCGG ATTATCTTTT GGGTGATGTT CCCCGCCCTT ttgCTTTcat 251 aCGTCTTTTC CGTTTTGTTC CACAACGGCA GTTGGACGGG CTTGGGCGCG 301 ctgTCCCAAT TCAACACCTT TGTCGGACGC ATCGCGCTGG CAAGTTTTGC 351 CGCCTACGCG CTCGGACAAA TCCTTGATAT TTTCGTATTC GACAAATTAC 401 GCCGTCTGAA AGCGTGGTGG ATTGCCCCGG CCGCATCAAC CGTCATCGGC 451 AATGCACTGG ACACGTTAGT ATTTTTTGCC GTTGCCTTTT ACGCAAGCAG 501 CGATGAATTT ATGGCGGCAA ACTGGCAGGG CATCGCTTTT GTCGATTACC 551 TGTTCAAACT TACCGTCTGC ACCCTCTTCT TCCTGCCCGC CTACGGCGTG 601 ATACTGAATC TGCTGACGAA AAAACTGACG GCCCTGCAAA CCAAACAGGC 651 GCAAGACCGC CCCGTGCCCT CGCTGCAAAA TCCGTAA This encodes a protein having amino acid sequence <SEQ LD 266>: 1 MYALTAAQQQ KALFRLVLFH ILIIAASΝYL VQFPFRIFGI HTTWGAFSFP 51 FIFLATDLTV RIFGSHLARR IIFWVMFPAL SLSYVFSVLF HΝGSWTGLGA 101 PSQFΝTFVGR IALASFAAYA LGQILDIFVF DKLRRLKAWW IAPAASTVIG 151 ΝALDTLVFFA VAFYASSDEF MAAΝWQGIAF VDYLFKLTVC TLFFLPAYGV 201 ILΝLLTKKLT ALQTKQAQDR PVPSLQΝP* An alternative annotated sequence is: 1 MYALTAAQQQ KALFRLVLFH ILIIAASΝYL VQFPFRIFGI HTTWGAFSFP 51 FIFLATDLTV RIFGSHLARR IIFWVMFPAL LLSYVFSVLF HΝGSWTGLGA 101 LSQFΝTFVGR IALASFAAYA LGQILDIFVF DKLRRLKAWW IAPAASTVIG 151 ΝALDTLVFFA VAFYASSDEF MAAΝWQGIAF VDYLFKLTVC TLFFLPAYGV 201 ILΝLLTKKLT ALQTKQAQDR PVPSLQΝP* ORF66ng and ORF66-1 show 96.1% identity in 228 aa overlap: orf66-1.pep MYAFTAAQQQKALFRLVLFHILIIAASNYLVQFPFQIFGIHTTWGAFSFPFIFLATDLTV 60 I I I: I II II I II I I II II II I II I I I II I II II II : II II I I I I II I I I II I II I I I I I I orf66ng MYALTAAQQQKALFRLVLFHILIIAASNYLVQFPFRIFGIHTTWGAFSFPFIFLATDLTV 60 orf66-1.pep RIFGSHLARRIIFWVMFPALLLSYVFSVLFHNGSWTGLGALSEFNTFVGRIALASFAAYA 120 II I II II I II II II I I II I II I I I II I I II II II II I II I II : II I I I I I I II II II II I orf66ng RIFGSHLARRIIFWVMFPALLLSYVFSVLFHNGSWTGLGALSQFNTFVGRIALASFAAYA 120 orf66-1. pep IGQILDIFVFNKLRRLKAWWIAPTASTVIGNALDTLVFFAVAFYASSDGFMAANWQGIAF 180 : I I II I I I II : I II I II II I II I : I II I II I I II I II I I I I I II II II II I II II I I I I orf66ng LGQILDIFVFDKLRRLKAWWIAPAASTVIGNALDTLVFFAVAFYASSDEFMAANWQGIAF 180 orf66-1.pep VDYLFKLTVCTLFFLPAYGVILNLLTKKLTTLQTKQAQDRPAPSLQNPX 229 II I II II I II II II I II II I II I II II I II : I I II II I II I : II II I II orf66ng VDYLFKLTVCTLFFLPAYGVILNLLTKKLTALQTKQAQDRPVPSLQNPX 229 Furthermore, ORF66ng shows significant homology with an E.coli ORF: sp|P37619|YHHQ_EC0LI HYPOTHETICAL 25.3 KD PROTEIN IN FTSY-NIKA INTERGENIC REGION (0221) >gi|1073495|pir||S47690 hypothetical protein o221 - Escherichia coli >gi|466607 (U00039) No definition line found [Escherichia coli] >gi 11789882 (AE000423) hypothetical 25.3 kD protein in ftsY-nikA intergenic region [Escherichia coli] Length = 221 Score = 273 bits (692), Expect = 5e-73 Identities = 132/203 (65%), Positives = 155/203 (76%) Query: 1 MYALTAAQQQKALFRLVLFHILIIAASNYLVQFPFRIFGIHTTWGAFSFPFIFLATDLTV 60 M + Q+ KALF L LFH+L+I +SNYLVQ P I G HTTWGAFSFPFIFLATDLTV Sbjct: 1 MNVFSQTQRYKALFWLSLFHLLVITSSNYLVQLPVSILGFHTTWGAFSFPFIFLATDLTV 60 Query: 61 RIFGSHLARRIIFWVMFPALLLSYVFSVLFHNGSWTGLGALSQFNTFVGRIALASFAAYA 120 RIFG+ LARRIIF VM PALL+SYV S LF+ GSW G GAL+ FN FV RIA ASF AYA Sbjct: 61 RIFGAPLARRIIFAVMIPALLISYVISSLFYMGSWQGFGALAHFNLFVARIATASFMAYA 120 Query: 121 LGQILDIFVFDKLRRLKAWWIAPAASTVIGNALDTLVFFAVAFYASSDEFMAANWQGIAF 180 LGQILD+ VF++LR+ + WW+AP AST+ GN DTL FF +AF+ S D FMA +W IA Sbjct: 121 LGQILDVHVFNRLRQSRRWWLAPTASTLFGNVSDTLAFFFIAFWRSPDAFMAEHWMEIAL 180 Query: 181 VDYLFKLTVCTLFFLPAYGVILN 203 VDY FK+ + +FFLP YGV+LN Sbjct: 181 VDYCFKVLISIVFFLPMYGVLLN 203 Based on this analysis, including the homology with the E.coli protein and the presence of several putative transmembrane domains in the gonococcal protein, it is predicted that these proteins from N. meningitidis .and N.gonorrhoeae, and their epitopes, could be useful .antigens for vaccines or diagnostics, or for raising antibodies. Example 32 The following partial DNA sequence was identified in N. meningitidis <SEQ ID 267>: 1 ATGGTCATAA AATATACAAA TTTGAATTTT GCGAAATTGT CGATAATTGC 51 AATTTTGATG ATGTATTCGT TTGAAGCGAA TGCAAAyGCA GT wrAATAT 101 CTGAAACTGT TTCAGTTGAT ACCGGACAAG GTGCGAAAAT TCATAAGTTT 151 GTACCTAAAA ATAGTAAAAC TTATTCATCT GATTTAATAA AAACGGTAGA 201 TTTAACACAC AyyCCTACGG GCGCAAAAGC CCGAATCAAC GCCAAAATAA 251 CCGCCAGCGT ATCCCGCGCC GGCGTATTGG CGGGGGTCGG CAAACTTGCC 301 CGCTTAGgCG CGAAATTCAG CACAAGGGCG GTtCCCTATG TCGGAACAGC 351 CcTTTTAGCC CACGACGTAT ACGAAAcTTT CAAAGAAGAC ATACAGGCAC 401 GAGGCTACCA ATACGACCCC GAAACCGACA AATTTGTAAA AGGCTACGAA 451 TATAGTAATT GCCTTTGGTA CGAAGACAAA AGACGTATTA ATAGAACCTA 501 TGGCTGCTAC GGCGTTGAT.. This corresponds to the amino acid sequence <SEQ ID 268; ORF72>: 1 MVIKYTNLNF AKLSIIAILM MYSFEANANA VXISETVSVD TGQGAKIHKF 51 VPKNSKTYSS DLIKTVDLTH XPTGAKARIN AKITASVSRA GVLAGVGKLA 101 RLGAKFSTRA VPYVGTALLA HDVYETFKED IQARGYQYDP ETDKFVKGYE 151 YSNCLWYEDK RRINRTYGCY GVD.. Further work revealed the complete nucleotide sequence <SEQ ID 269>: 1 ATGGTCATAA AATATACAAA TTTGAATTTT GCGAAATTGT CGATAATTGC 51 AATTTTGATG ATGTATTCGT TTGAAGCGAA TGCAAATGCA GTAAAAATAT 101 CTGAAACTGT TTCAGTTGAT ACCGGACAAG GTGCGAAAAT TCATAAGTTT 151 GTACCTAAAA ATAGTAAAAC TTATTCATCT GATTTAATAA AAACGGTAGA 201 TTTAACACAC ATCCCTACGG GCGCAAAAGC CCGAATCAAC GCCAAAATAA 251 CCGCCAGCGT ATCCCGCGCC GGCGTATTGG CGGGGGTCGG CAAACTTGCC 301 CGCTTAGGCG CGAAATTCAG CACAAGGGCG GTTCCCTATG TCGGAACAGC 351 CCTTTTAGCC CACGACGTAT ACGAAACTTT CAAAGAAGAC ATACAGGCAC 401 GAGGCTACCA ATACGACCCC GAAACCGACA AATTTGCAAA GGTCTCAGGC 451 TAA This corresponds to the amino acid sequence <SEQ ID 270; ORF72-l>: 1 MVIKYTNLNF AKLSIIAILM MYSFEANANA VKISETVSVD TGQGAKIHKF 51 VPKNSKTYSS DLIKTVDLTH IPTGAKARIN AKITASVSRA GVLAGVGKLA 101 RLGAKFSTRA VPYVGTALLA HDVYETFKED IQARGYQYDP ETDKFAKVSG 151 * Computer analysis of this amino acid sequence gave the following results: Homology with a predicted ORF from N.meninsitidis (strain A) ORF72 shows 98.0% identity over a 147aa overlap with an ORF (ORF72a) from strain A of N. meningitidis: 10 20 30 40 50 60 orf72. pep MVIKYTNLNFAKLSIIAILMMYSFEANANAVXISETVSVDTGQGAKIHKFVPKNSKTYSS I I II I II I I II I I II I I I II II I II I II I II II I I I II I I I I II II I I I I II II I I I I I orf72a MVIKYTNLNFAKLSIIAILMMYSFEANANAVKISETVSVDTGQGAKIHKFVPKNSKTYSS 10 20 30 40 50 60 70 80 90 100 110 120 orf72.pep DLIKTVDLTHXPTGAKARINAKITASVSRAGVLAGVGKLARLGAKFSTRAVPYVGTALLA I II I I II I II II I I I II I I II I II I II II I I II II I I I I I I I II I II II II II II II I I orf72a DLIKTVDLTHIPTGAKARINAKITASVSRAGVLAGVGKLARLGAKFSTRAVPYVGTALLA 70 80 90 100 110 120 130 140 150 160 170 orf72.pep HDVYETFKEDIQARGYQYDPETDKFVKGYEYSNCLWYEDKRRINRTYGCYGVD I I I II I II I II I I I I I I II II I II I: I orf72a HDVYETFKEDIQARGYQYDPETDKFAKVSGX 130 140 150 The complete length ORF72a nucleotide sequence <SEQ ID 271> is: 1 ATGGTCATAA AATATACAAA TTTGAATTTT GCGAAATTGT CGATAATTGC 51 AATTTTGATG ATGTATTCGT TTGAAGCGAA TGCAAATGCA GTAAAAATAT 101 CTGAAACTGT TTCAGTTGAT ACCGGACAAG GTGCGAAAAT TCATAAGTTT 151 GTACCTAAAA ATAGTAAAAC TTATTCATCT GATTTAATAA AAACGGTAGA 201 TTTAACACAC ATCCCTACGG GCGCAAAAGC CCGAATCAAC GCCAAAATAA 251 CCGCCAGCGT ATCCCGCGCC GGCGTATTGG CGGGGGTCGG CAAACTTGCC 301 CGCTTAGGCG CGAAATTCAG CACAAGGGCG GTTCCCTATG TCGGAACAGC 351 CCTTTTAGCC CACGACGTAT ACGAAACTTT CAAAGAAGAC ATACAGGCAC 401 GAGGCTACCA ATACGACCCC GAAACCGACA AATTTGCAAA GGTCTCAGGC 451 TAA This encodes a protein having amino acid sequence <SEQ ID 272>: 1 MVIKYTNLNF AKLSIIAILM MYSFEANANA VKISETVSVD TGQGAKIHKF 51 VPKNSKTYSS DLIKTVDLTH IPTGAKARIN AKITASVSRA GVLAGVGKLA 101 RLGAKFSTRA VPYVGTALLA HDVYETFKED IQARGYQYDP ETDKFAKVSG 151 * ORF72a and ORF72-1 show 100.0% identity in 150 aa overlap: 10 20 30 40 50 60 orf72a. pep MVIKYTNLNFAKLSIIAILMMYSFEANANAVKISETVSVDTGQGAKIHKFVPKNSKTYSS I I I I I II I I I II II II I I I MM II II I I I II I I II II II II I II M M I II II I I I II I orf72-1 MVIKYTNLNFAKLSIIAILMMYSFEANANAVKISETVSVDTGQGAKIHKFVPKNSKTYSS 10 20 30 40 50 60 70 80 90 100 110 120 orf72 . pep DLIKTVDLTHIPTGAKARINAKITASVSRAGVLAGVGKLARLGAKFSTRAVPYVGTALLA I II I I II II I I I II II I II I II I II I II II I II I II I II I II II II I I I I II I II II II I orf72-1 DLIKTVDLTHIPTGAKARINAKITASVSRAGVLAGVGKLARLGAKFSTRAVPYVGTALLA 70 80 90 100 110 120 130 140 150 orf72a . pep HDVYETFKEDIQARGYQYDPETDKFAKVSGX I II II I I I I I II II II II I I I II I I I I II II orf72-1 HDVYETFKEDIQARGYQYDPETDKFAKVSGX 130 140 150 Homology with a predicted ORF from N.sonorrhoeae ORF72 shows 89% identity over a 173aa overlap with a predicted ORF (ORF72.ng) from N. gonorrhoeae: orf 2.pep MVIKYTNLNFAKLSIIAILMMYSFEANANAVXISETVSVDTGQGAKIHKFVPKNSKTYSS 60 II I: I I II I II II II I I II I I II I I I II II I II I: I II I II I I I : II II II: M III orf72ng MVTKHTNLNFAKLSIIAILMMYSFEANANAVKISETLSVDTGQGAKVHKFVPKSSNIYSS 60 orf72. pep DLIKTVDLTHXPTGAKARINAKITASVSRAGVLAGVGKLARLGAKFSTRAVPYVGTALLA 120 II I: I I II I I II II I I II I I II I II I II II I: I I II IM I I I I: I I I I II I I I I II I orf72ng DLTKAVDLTHIPTGAKARINAKITASVSRAGVLSGVGKLVRQGAKFGTRAVPYVGTALLA 120 orf72.pep HDVYETFKEDIQARGYQYDPETDKFVKGYEYSNCLWYEDKRRINRTYGCYGVD 173 II II I I II I I II II I : I II I I II I II I II I : I II II II : II I I I II I I I I II orf72ng HDVYETFKEDIQARGCRYDPETDKFVKGYEYANCLWYEDERRINRTYGCYGVDSSIMRLM 180 An ORF72ng nucleotide sequence <SEQ ED 273> was predicted to encode a protein having amino acid sequence <SEQ ID 274>: 1 MVTKHTNLNF AKLSIIAILM MYSFEANANA VKISETLSVD TGQGAKVHKF 51 VPKSSNIYSS DLTKAVDLTH IPTGAKARIN AKITASVSRA GVLSGVGKLV 101 RQGAKFGTRA VPYVGTALLA HDVYETFKED IQARGCRYDP ETDKFVKGYE 151 YANCLWYEDE RRINRTYGCY GVDSSIMRLM PDRSRFPEVK QLMESQMYRL 201 ARPFWNWRKE ELNKLSSLDW NNFVLNRCTF DWNGGGCAVN KGDDFRAGAS 251 FSLGRNPKYK EEMDAKKPEE ILSLKVDADP DKYIEATGYP GYSEKVEVAP 301 GTKVNMGPVT DRNGNPVQVA ATFGRDAQGN TTADVQVIPR PDLTPASAEA 351 PHAQPLPEVS PAENPANNPD PDENPGTRPN PEPDPDLNPD ANPDTDGQPG 401 TSPDSPAVPD RPNGRHRKER KEGEDGGLSC DYFPEILACQ EMGKPSDRMF 451 HDISIPQVTD DKTWSSHNFL PSNGVCPQPK TFHVFGRQYR ASYEPLCVFA 501 EKIRFAVLLA FIIMSAFWF GSLGGE* After further analysis, the following gonococcal DNA sequence <SEQ ID 275> was identified: 1 ATGGTCACAA AACATACAAA TTTGAATTTT GCGAAATTGT CGATAATTGC 51 AATTTTGATG ATGTATTCGT TTGAAGCGAA TGCAAATGCA GTAAAAATAT 101 CTGAAACTCT TTCGGTTGAT ACCGGACAAG GCGCGAAAGT TCATAAGTTC 151 GTTCCTAAAT CAAGTAATAT TTATTCATCT GATTTAACAA AAGCGGTAGA 201 TTTAACGCAT ATCCCCACGG GCGCAAAAGC CCGAATCAAC GCCAAAATAA 251 CCGCCAGCGT ATCCCGCGCC GGCGTATTGT CGGGGGTCGG CAAACTTGTC 301 CGCCAAGGCG CGAAATTCGG CACAAGGGCG GTTCCCTATG TCGGAACAGC 351 CCTTTTAGCC CACGACGTAT ACGAAACTTT CAAAGAAGAC ATACAGGCAC 401 GAGGCTGCCG ATACGATCCC GAAACCGACA AATTT This corresponds to the .amino acid sequence <SEQ ID 276; ORF72ng-l>: 1 MVTKHTNLNF AKLSIIAILM MYSFEANANA VKISETLSVD TGQGAKVHKF 51 VPKSSNIYSS DLTKAVDLTH IPTGAKARIN AKITASVSRA GVLSGVGKLV 101 RQGAKFGTRA VPYVGTALLA HDVYETFKED IQARGCRYDP ETDKF ORF72ng-l and ORF721-1 show 89.7% identity in 145 aa overlap: 10 20 30 40 50 60 orf72ng-l.pe MVTKHTNLNFAKLSIIAILMMYSFEANANAVKISETLSVDTGQGAKVHKFVPKSSNIYSS II I : II II II I I II II I II II I II II I II II II II : II II I I II I : I I II II: I: III orf72-1 MVIKYTNLNFAKLSIIAILMMYSFEANANAVKISETVSVDTGQGAKIHKFVPKNSKTYSS 10 20 30 40 50 60 70 80 90 100 110 120 orf72ng-l.pe DLTKAVDLTHIPTGAKARINAKITASVSRAGVLSGVGKLVRQGAKFGTRAVPYVGTALLA II M I I I II II I I II I II II II I II II I II II: I I II I : I I I II : II I II I I I II I II orf72-1 DLIKTVDLTHIPTGAKARINAKITASVSRAGVLAGVGKLARLGAKFSTRAVPYVGTALLA 70 80 90 100 110 120 130 140 orf72ng-l.pe HDVYETFKEDIQARGCRYDPETDKF II II I II II II I I I I : II II II I I orf 2-1 HDVYETFKEDIQARGYQYDPETDKFAKVSGX 130 140 150 Based on this analysis, including the presence of a putative leader sequence and transmembrane domains in the gonococcal protein, it is predicted that the proteins from N. meningitidis and N.gonorrhoeae, and their epitopes, could be useful antigens for vaccines or diagnostics, or for raising antibodies. Example 33 The following partial DNA sequence was identified in N. meningitidis <SEQ ID 277>: 1 ATGAGATTTT TCGGTATCGG TTTTTTGGTG CTGCTGTTTT TGGAGATTAT 51 GTCGATTGTG TGGGTTGCCG ATTGGCTGGG CGGCGGCTGG ACGTTGTTTT 101 TGATGGCGGC AGGTTTTGCC GCCGGCGTGC TGATGCTCAG GCAAACCGGG 151 gCTGACCGGT CTTTTATTGG CGGGCGCGGC AATGAGAAGC GGCGGGAAGG 201 TATCCGTTTA TCAGATGTTG TGGCCTATC .. This corresponds to the amino acid sequence <SEQ ID 278; ORF73>: 1 MRFFGIGFLV LLFLEIMSIV WVADWLGGGW TLFLMAAGFA AGVLMLRQTG 51 LTGLLLAGAA MRSGGKVSVY QMLWPI .. Further work revealed the complete nucleotide sequence <SEQ ID 279>: 1 ATGAGATTTT TCGGTATCGG TTTTTTGGTG CTGCTGTTTT TGGAGATTAT 51 GTCGATTGTG TGGGTTGCCG ATTGGCTGGG CGGCGGCTGG ACGTTGTTTT 101 TGATGGCGGC AGGTTTTGCC GCCGGCGTGC TGATGCTCAG GCATACGGGG 151 CTGTCCGGTC TTTTATTGGC GGGCGCGGCA ATGAGAAGCG GCGGGAGGGT 201 ATCCGTTTAT CAGATGTTGT GGCCTATCCG TTATACGGTG GCGGCTGTGT 251 GTCTGATGAG TCCGGGATTC GTATCCTCGG TGTTGGCGGT ATTGCTGCTG 301 CTGCCGTTTA AGGGAGGGGC AGTGTTGCAG GCAGGAGGTG CGGAAAATTT 351 TTTCAACATG AACCAATCGG GCAGAAAAGA GGGCTTTTCC CGCGATGACG 401 ATATTATCGA GGGAGAATAT ACGGTTGAAG AGCCTTACGG CGGCAATCGT 451 TCCCGAAACG CCATCGAACA CAAAAAAGAC GAATAA This corresponds to the amino acid sequence <SEQ ID 280; ORF73-l>: 1 MRFFGIGFLV LLFLEIMSIV WVADWLGGGW TLFLMAAGFA AGVLMLRHTG 51 LSGLLLAGAA MRSGGRVSVY QMLWPIRYTV AAVCLMSPGF VSSVLAVLLL 101 LPFKGGAVLQ AGGAENFFNM NQSGRKEGFS RDDDIIEGEY TVEEPYGGNR 151 SRNAIEHKKD E* Computer analysis of this amino acid sequence gave the following results: Homology with a predicted ORF from N.meninsitidis (strain Al ORF73 shows 90.8% identity over a 76aa overlap with an ORF (ORF73a) from strain A of N. meningitidis: 10 20 30 40 50 60 orf73. pep MRFFGIGFLVLLFLEIMSIVWVADWLGGGWTLFLMAAGFAAGVLMLRQTGLTGLLLAGAA II I I II I II II I II I II II II I I II II I II II II I II I I I II : I I I : I I I : I II I I I I I orf73a MRFFGIGFLVLLFLEIMSIVWVADWLGGGWTLFLMAATFAAGWMLRHTGLSGLLLAGAA 10 20 30 40 50 60 70 orf73.pep MRSGGKVSVYQMLWPI I I II I : I I II III I orf73a MRSGGRVSVYXMLWXIRYTVAAVCXMSPGFVSSVXAVLLXLPFKGGAVLQAGGAENFFNM The complete length ORF73a nucleotide sequence <SEQ ID 281> is: 1 ATGAGATTTT TCGGTATCGG TTTTTTGGTG CTGCTGTTTT TGGAGATTAT 51 GTCGATTGTG TGGGTTGCCG ATTGGTTGGG CGGCGGTTGG ACGCTGTTTC 101 TAATGGCGGC AACCTTTGCC GCCGGCGTGG TGATGCTCAG GCATACGGGG 151 CTGTCCGGTC TTTTATTGGC GGGCGCGGCA ATGAGAAGCG GCGGGAGGGT 201 ATCCGTTTAT CANATGTTGT GGCNTATCCG TTATACGGTG GCGGCGGTGT 251 GTCNGATGAG TCCGGGATTC GTATCCTCGG TGTNGGCGGT ATTGCTGNTG 301 CTNCCGTTTA AGGGAGGTGC AGTGTTGCAG GCAGGAGGTG CGGAAAATTT 351 TTTCAACATG AACCANTCGG GCAGAAAAGA NGGCNTTTCC CGCGATGACG 401 ATATTATCGA GGGGGAATAT ACGGTTGAAG ANCCTTACGG CGGCANTCGT 451 TTCCGAAACG CCNTNGAACA CAAAAAAGAC GAATAA This encodes a protein having amino acid sequence <SEQ ID 282>: 1 MRFFGIGFLV LLFLEIMSIV WVADWLGGGW TLFLMAATFA AGWMLRHTG 51 LSGLLLAGAA MRSGGRVSVY XMLWXIRYTV AAVCXMSPGF VSSVXAVLLX 101 LPFKGGAVLQ AGGAENFFNM NXSGRKXGXS RDDDIIEGEY TVEXPYGGXR 151 FRNAXEHKKD E* ORF73a and ORF73-1 show 91.3% identity in 161 aa overlap 10 20 30 40 50 60 orf73a . pep MRFFGIGFLVLLFLEIMSIVWVADWLGGGWTLFLMAATFAAGWMLRHTGLSGLLLAGAA II I I II I II I II I I II I II I II I I II I I I II II I I I I II I II : I II I I I I I II I II II I orf73-1 MRFFGIGFLVLLFLEIMSIVWVADWLGGGWTLFLMAAGFAAGVLMLRHTGLSGLLLAGAA 10 20 30 40 50 60 70 80 90 100 110 120 orf73a . pep MRSGGRVSVYXMLWXIRYTVAAVCXMSPGFVSSVXAVLLXLPFKGGAVLQAGGAENFFNM II I I II I I I I III II II I I II I II II I II II II II II I II I I II II I II I I I I I I orf 3-1 MRSGGRVSVYQMLWPIRYTVAAVCLMSPGFVSSVLAVLLLLPFKGGAVLQAGGAENFFNM 70 80 90 100 110 120 130 140 150 160 orf73a. pep NXSGRKXGXSRDDDIIEGEYTVEXPYGGXRFRNAXEHKKDEX I I I II I I II II I I II I II I I I I II I III II I I I II orf73-1 NQSGRKEGFSRDDDIIEGEYTVEEPYGGNRSRNAIEHKKDEX 130 140 150 160 Homology with a predicted ORF from Nsonorrhoeae ORF73 shows 92.1% identity over a 76aa overlap with a predicted ORF (ORF73.ng) from N gonorrhoeae: orf 73 . pep MRFFGIGFLVLLFLEIMSIVWVADWLGGGWTLFLMAAGFAAGVLMLRQTGLTGLLLAGAA 60 I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I M M I I : I I I I I I I I orf73ng MRFFGIGFLVLLFLEIMSIVWVADWLGGGWTLFLMAATFAAGVLMLRHTGLSGLLLAGAA 60 orf73. pep MRSGGKVSVYQMLWPI 76 :: I : I I II II I I II II orf73ng VKSSGKVSVYQMLWPIRYTVAAVCLMSPGFVSSVLAVLLLLPFKGGAVLQAGGAENFFNM 120 The complete length ORF73ng nucleotide sequence <SEQ ID 283> is: 1 ATGAGATTTT TCGGTATCGG TTTTTTGGTG CTGCTGTTTT TGGAAATTAT 51 GTCGATTGTG TGGGTTGCCG ATTGGCTGGG CGGCGGTTGG AcgcTGTTTC 101 TAATGGCGGC AACCTTTGCC GCCGGTGTGC TGATGCTCAG GCATAcggGG 151 CTGTCCGGTC TTTTATTGGC TGGCGCGGCG GTAAAAagta gtgGGAAGGT 201 ATCTGTTTAT CagatgtTGT GGCCTATCCG TTATAcggtg gcggcggtgT 251 GTCTGatgag tCcggGATTC GTATCCTccg tgttggCGGT ATTGCTGCTG 301 CTGCcgttta aggGaggGgc agtgttgcag gcaggaggtg cggaaaATTT 351 TTTCAACATg aaCcaatcgg gcagaaAaga gggatttttc cacgatgacg 401 atattatcga gggagaatat acggttgaaa aacctgacgg cggcaatcgt 451 tcccgaAAcg ccatcgaaca cgaaaAagac gaataA This encodes a protein having amino acid sequence <SEQ ID 284>: 1 MRFFGIGFLV LLFLEIMSIV WVADWLGGGW TLFLMAATFA AGVLMLRHTG 51 LSGLLLAGAA VKSSGKVSVY QMLWPIRYTV AAVCLMSPGF VSSVLAVLLL 101 LPFKGGAVLQ AGGAENFFNM NQSGRKEGFF HDDDIIEGEY TVEKPDGGNR 151 SRNAIEHEKD E* ORF73ng and ORG73-1 show 93.8% identity in 161 aa overlap 10 20 30 40 50 60 orf73-1. pep MRFFGIGFLVLLFLEIMSIVWVADWLGGGWTLFLMAAGFAAGVLMLRHTGLSGLLLAGAA I II II II I I II II II II II I II I I II I I II I II I II I II II I II I I I II II I I I II I I I orf73ng MRFFGIGFLVLLFLEIMSIVWVADWLGGGWTLFLMAATFAAGVLMLRHTGLSGLLLAGAA 10 20 30 40 50 60 70 80 90 100 110 120 orf73-1.pep MRSGGRVSVYQMLWPIRYTVAAVCLMSPGFVSSVLAVLLLLPFKGGAVLQAGGAENFFNM :: I : I : II II I II II II I I I I II I I II I II I II I II II II II I I II II II I II I I I II I I orf73ng VKSSGKVSVYQMLWPIRYTVAAVCLMSPGFVSSVLAVLLLLPFKGGAVLQAGGAENFFNM 70 80 90 100 110 120 130 140 150 160 orf73-1.pep NQSGRKEGFSRDDDIIEGEYTVEEPYGGNRSRNAIEHKKDEX II I II II I I : I II II I II I II I : I I I II II I I I I I : II II orf73ng NQSGRKEGFFHDDDIIEGEYTVEKPDGGNRSRNAIEHEKDEX 130 140 150 160 Based on this analysis, including the presence of a putative leader sequence and putative transmembrane domain in the gonococcal protein, it is predicted that the proteins from N. meningitidis and N.gonorrhoeae, and their epitopes, could be useful antigens for vaccines or diagnostics, or for raising antibodies. Example 34 The following partial DNA sequence was identified in N. meningitidis <SEQ ID 285>: 1 ATGTTTGTTT TTCAGACGGC ATTCTT.ATG TTTCAGAAAC ATTTGCAGAA 51 AGCCTCCGAC AGCGTCGTCG GAGGGACATT ATACGTGGTT GCCACGCCCA 101 TCGGCAATTT GGCGGACATT ACCCTGCGCG CTTTGGCGGT ATTGCAAAAG 151 GCG GCCGA AGACACGCGC GTTACCGCAC AGCTTTTGAG 201 CGCGTACGGC ATTCAGGGCA AACTCGTCAG TGTGCGCGAA CACAACGAAC 251 GGCAGATGGC GGACAAGATT GTCGGCTATC TTTCAGACGG CATGGTTGTG 301 GCACAGGTTT CCGATGCGGG TACGCCGGCC GTGTGCGACC CGGGCGCGAA 351 ACTCGCCCGC CGCGTGCGTG AGGCCGGGTT TAAAGTCGTT CCCGTCGTGG 401 GCGCAAC.GC GGTGATGGCG GCTTTGAGCG TGGCCGGTGT GGAAGGATCC 451 GATTTTTATT TCAACGGTTT TGTACCGCCG AAATCGGGAG AACGCAGGAA 501 ACTGTTTGCC AAATGGGTGC GGGCGGCGTT TCCTATCGTC ATGTTTGAAA 551 CGCCGCACCG CATCGGTGCA GCGCTTGCCG ATATGGCGGA ACTGTTCCCC 601 GAACGCCGAT TAATGCTGGC GCGCGAAATT ACGAAAACGT TTGAAACGTT 651 CTTAAGCGGC ACGGTTGGGG AAATTCAGAC GGCATTGTCT GCCGACGGCG 701 ACCAATCGCG CGGCGAGATG GTGTTGGTGC TTTATCCGGC GCAGGATGAA 751 AAACACGAAG GCTTGTCCGA GTCCGCGCAA AACATCATGA AAATCCTCAC 801 AGCCGAGCTG CCGACCAAAC AGGCGGCGGA GCTTGCTGCC AAAATCACGG 851 GCGAGGGAAA GAAAGCTTTG TACGAT.. This corresponds to the amino acid sequence <SEQ ID 286; ORF75>: 1 MFVFQTAFXM FQKHLQKASD SWGGTLYW ATPIGNLADI TLRALAVLQK 51 A....AEDTR VTAQLLSAYG IQGKLVSVRE HNERQMADKI VGYLSDGMW 101 AQVSDAGTPA VCDPGAKLAR RVREAGFKW PWGAXAVMA ALSVAGVEGS 151 DFYFNGFVPP KSGERRKLFA KWVRAAFPIV MFETPHRIGA ALADMAELFP 201 ERRLMLAREI TKTFETFLSG TVGEIQTALS ADGDQSRGEM VLVLYPAQDE 251 KHEGLSESAQ NIMKILTAEL PTKQAAELAA KITGEGKKAL YD.. Further work revealed the complete nucleotide sequence <SEQ ID 287>: 1 ATGTTTCAGA AACATTTGCA GAAAGCCTCC GACAGCGTCG TCGGAGGGAC 51 ATTATACGTG GTTGCCACGC CCATCGGCAA TTTGGCGGAC ATTACCCTGC 101 GCGCTTTGGC GGTATTGCAA AAGGCGGACA TCATCTGTGC CGAAGACACG 151 CGCGTTACCG CACAGCTTTT GAGCGCGTAC GGCATTCAGG GCAAACTCGT 201 CAGTGTGCGC GAACACAACG AACGGCAGAT GGCGGACAAG ATTGTCGGCT 251 ATCTTTCAGA CGGCATGGTT GTGGCACAGG TTTCCGATGC GGGTACGCCG 301 GCCGTGTGCG ACCCGGGCGC GAAACTCGCC CGCCGCGTGC GTGAGGCCGG 351 GTTTAAAGTC GTTCCCGTCG TGGGCGCAAG CGCGGTGATG GCGGCTTTGA 401 GCGTGGCCGG TGTGGAAGGA TCCGATTTTT ATTTCAACGG TTTTGTACCG 451 CCGAAATCGG GAGAACGCAG GAAACTGTTT GCCAAATGGG TGCGGGCGGC 501 GTTTCCTATC GTCATGTTTG AAACGCCGCA CCGCATCGGT GCGACGCTTG 551 CCGATATGGC GGAACTGTTC CCCGAACGCC GATTAATGCT GGCGCGCGAA 601 ATTACGAAAA CGTTTGAAAC GTTCTTAAGC GGCACGGTTG GGGAAATTCA 651 GACGGCATTG TCTGCCGACG GCAACCAATC GCGCGGCGAG ATGGTGTTGG 701 TGCTTTATCC GGCGCAGGAT GAAAAACACG AAGGCTTGTC CGAGTCCGCG 751 CAAAACATCA TGAAAATCCT CACAGCCGAG CTGCCGACCA AACAGGCGGC 801 GGAGCTTGCT GCCAAAATCA CGGGCGAGGG AAAGAAAGCT TTGTACGATC 851 TGGCTCTGTC TTGGAAAAAC AAATAG This corresponds to the amino acid sequence <SEQ ID 288; ORF75-l>: 1 MFQKHLQKAS DSWGGTLYV VATPIGNLAD ITLRALAVLQ KADIICAEDT 51 RVTAQLLSAY GIQGKLVSVR EHNERQMADK IVGYLSDGMV VAQVSDAGTP 101 AVCDPGAKLA RRVREAGFKV VPWGASAVM AALSVAGVEG SDFYFNGFVP 151 PKSGERRKLF AKWVRAAFPI VMFETPHRIG ATLADMAELF PERRLMLARE 201 ITKTFETFLS GTVGEIQTAL SADGNQSRGE MVLVLYPAQD EKHEGLSESA 251 QNIMKILTAE LPTKQAAELA AKITGEGKKA LYDLALSWKN K* Computer analysis of this amino acid sequence gave the following results: Homology with a predicted ORF from N. meningitidis (strain A) ORF75 shows 95.8% identity over a 283aa overlap with an ORF (ORF75a) from strain A of N. meningitidis: 10 20 30 40 50 60 orf75.pep MFVFQTAFXMFQKHLQKASDSWGGTLYWATPIGNLADITLRALAVLQKAXXXXAEDTR I II II I I I I I II II I I I I I I I II I II I I I I I I II I II I I II I I I I I I orf75a MFQKHLQKASDSVVGGTLYWATPIGNLADITLRALAVLQKADIICAEDTR 10 20 30 40 50 70 80 90 100 110 120 orf75.pep VTAQLLSAYGIQGKLVSVREHNERQMADKIVGYLSDGMWAQVSDAGTPAVCDPGAKLAR II I I II I II I II II II I I I I II II I I I I I I II I I I II I I I I I II I II II I I II I I I I I I I orf75a VTAQLLSAYGIQGKLVSVREHNERQMADKIVGYLSDGMWAQVSDAGTPAVCDPGAKLAR 60 70 80 90 100 110 130 140 150 160 170 180 orf75.pep RVREAGFKWPWGAXAVMAALSVAGVEGSDFYFNGFVPPKSGERRKLFAKWVRAAFPIV II I I : I II I I I I II I I II I I I II II I I II II I II II II I II I I I I I I II I I I : I I I: I orf 75a RVREVGFKWP GASAVMAALSVAGVAGSDFYFNGFVPPKSGERRKLFAKWVRVAFPW 120 130 140 150 160 170 190 200 210 220 230 240 orf75.pep MFETPHRIGAALADMAELFPERRLMLAREITKTFETFLSGTVGEIQTALSADGDQSRGEM II II I II I II : I II II II I II I II II I II I I I I I II II I I I I II I I I II : I I I : II I II I orf75a MFETPHRIGATLADMAELFPERRLMLAREITKTFETFLSGTVGEIQTALAADGNQSRGEM 180 190 200 210 220 230 250 260 270 280 290 orf75. pep VLVLYPAQDEKHEGLSESAQNIMKILTAELPTKQAAELAAKITGEGKKALYD II I II I II I II II II II I II I II I II II I II II I II II II I II II I II I I I I orf75a VLVLYPAQDEKHEGLSESAQNIMKILTAELPTKQAAELAAKITGEGKKALYDLALSWKNK 240 250 260 270 280 290 orf75a X The complete length ORF75a nucleotide sequence <SEQ LD 289> is: 1 ATGTTTCAGA AACATTTGCA GAAAGCCTCC GACAGCGTCG TCGGAGGGAC 51 ATTATACGTG GTTGCCACGC CCATCGGCAA TTTGGCGGAC ATTACCCTGC 101 GCGCTTTGGC GGTATTGCAA AAGGCGGACA TCATCTGTGC CGAAGACACG 151 CGCGTTACCG CGCAGCTTTT GAGCGCGTAC GGCATTCAGG GCAAACTCGT 201 CAGCGTGCGC GAACACAACG AACGGCAGAT GGCGGACAAG ATTGTCGGCT 251 ATCTTTCAGA CGGCATGGTT GTGGCACAGG TTTCCGATGC GGGTACGCCG 301 GCCGTGTGCG ACCCGGGCGC GAAACTCGCC CGCCGCGTGC GTGAGGTCGG 351 GTTTAAAGTT GTCCCTGTTG TCGGCGCAAG CGCGGTGATG GCGGCTTTGA 401 GTGTGGCTGG TGTGGCGGGA TCCGATTTTT ATTTCAACGG TTTTGTACCG 451 CCGAAATCGG GCGAACGTAG GAAATTGTTT GCCAAATGGG TGCGGGTGGC 501 GTTTCCCGTC GTGATGTTTG AAACGCCGCA CCGCATCGGG GCGACGCTTG 551 CCGATATGGC GGAACTGTTC CCCGAACGCC GATTAATGCT GGCGCGCGAA 601 ATCACGAAAA CGTTTGAAAC GTTCTTAAGC GGCACGGTTG GGGAAATTCA 651 GACGGCATTG GCGGCGGACG GCAACCAATC GCGCGGCGAG ATGGTGTTGG 701 TGCTTTATCC GGCGCAGGAT GAAAAACACG AAGGCTTGTC CGAGTCCGCG 751 CAAAACATCA TGAAAATCCT CACAGCCGAG CTGCCGACCA AACAGGCGGC 801 GGAGCTTGCC GCCAAAATCA CGGGCGAGGG AAAAAAAGCT TTGTACGATC 851 TGGCACTGTC TTGGAAAAAC AAATGA This encodes a protein having amino acid sequence <SEQ ID 290>: 1 MFQKHLQKAS DSWGGTLYV VATPIGNLAD ITLRALAVLQ KADIICAEDT 51 RVTAQLLSAY GIQGKLVSVR EHNERQMADK IVGYLSDGMV VAQVSDAGTP 101 AVCDPGAKLA RRVREVGFKV_ VPWGASAVM AALSVAGVAG SDFYFNGFVP 151 PKSGERRKLF AKWVRVAFPV VMFETPHRIG ATLADMAELF PERRLMLARE 201 ITKTFETFLS GTVGEIQTAL AADGNQSRGE MVLVLYPAQD EKHEGLSESA 251 QNIMKILTAE LPTKQAAELA AKITGEGKKA LYDLALSWKN K* ORF75a and ORF75-1 show 98.3% identity in 291 aa overlap: 10 20 30 40 50 60 orf 75a . pep MFQKHLQKASDSWGGTLYWATPIGNLADITLRALAVLQKADIICAEDTRVTAQLLSAY II I I II I II II I II I I II I II I II I I II I II II I II II I I II I II I II II II II II I II I orf75-1 MFQKHLQKASDSWGGTLYWATPIGNLADITLRALAVLQKADIICAEDTRVTAQLLSAY 10 20 30 40 50 60 70 80 90 100 110 120 orf 5a. pep GIQGKLVSVREHNERQMADKIVGYLSDGMWAQVSDAGTPAVCDPGAKLARRVREVGFKV II I II I II I II I II I I II I II I I II I I II II II I II I II I I I II II I II I I II II: I I I I orf 75-1 GIQGKLVSVREHNERQMADKIVGYLSDGMWAQVSDAGTPAVCDPGAKLARRVREAGFKV 70 80 90 100 110 120 130 140 150 160 170 180 orf 75a . pep VPWGASAVMAALSVAGVAGSDFYFNGFVPPKSGERRKLFAKWVRVAFPWMFETPHRIG I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I : I I I : I I I I I I I I I I orf 75-1 VPWGASAVMAALSVAGVEGSDFYFNGFVPPKSGERRKLFAKWVRAAFPIVMFETPHRIG 130 140 150 160 170 180 190 200 210 220 230 240 orf 75a . pep m ATLADMAELFPERRLMLAREITKTFETFLSGTVGEIQTALAADGNQSRGEMVLVLYPAQD I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I : I I I I I I I I I I I I I I I I I I I orf75-1 ATLADMAELFPERRLMLAREITKTFETFLSGTVGEIQTALSADGNQSRGEMVLVLYPAQD 190 200 210 220 230 240 250 260 270 280 290 orf 5a . pep EKHEGLSESAQNIMKILTAELPTKQAAELAAKITGEGKKALYDLALSWKNKX II II I I I I I II I I II I II I I II I II I II II II I II II II II II II II I I I II orf75-1 EKHEGLSESAQNIMKILTAELPTKQAAELAAKITGEGKKALYDLALSWKNKX 250 260 270 280 290 Homology with a predicted ORF from N.sonorrhoeae ORF75 shows 93.2% identity over a 292aa overlap with a predicted ORF (ORF75.ng) from N. gonorrhoeae: orf 75.pep MFVFQTAFXMFQKHLQKASDSWGGTLYVVATPIGNLADITLRALAVLQKA AEDTR 56 I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I orf75ng MSVFQTAFFMFQKHLQKASDSWGGTLYWATPIGNLADITLRALAVLQKADIICAEDTR 60 orf75.pep VTAQLLSAYGIQGKLVSVREHNERQMADKIVGYLSDGMWAQVSDAGTPAVCDPGAKLAR 116 I I I I I I I I I I I I I : I I I I I I I I I I I I I I M: I : I I I I : I I I I I I I I I I I I I I I I I I I I I I orf 75ng VTAQLLSAYGIQGRLVSVREHNERQMADKVIGFLSDGLWAQVSDAGTPAVCDPGAKLAR 120 orf75.pep RVREAGFKWPWGAXAVMAALSVAGVEGSDFYFNGFVPPKSGERRKLFAKWVRAAFPIV 176 I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I: I orf75ng RVREAGFKVVPWGASAVMAALSVAGVAESDFYFNGFVPPKSGERRKLFAKWVRAAFPW 180 orf 75.pep MFETPHRIGAALADMAELFPERRLMLAREITKTFETFLSGTVGEIQTALSADGDQSRGEM 236 I I I I I I I I I I : I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I : I I I : I I I I I I orf75ng MFETPHRIGATLADMAELFPERRLMLAREITKTFETFLSGTVGEIQTALAADGNQSRGEM 240 orf 75.pep VLVLYPAQDEKHEGLSESAQNIMKILTAELPTKQAAELAAKITGEGKKALYD 288 I I I I I I I I I I I I I I I I I I I I I I I I I: I I I I I I I I I I I I I I I I I I I I I I I I I o urϋf7 /5 -jniigy 3 JU00U An ORF75ng nucleotide sequence <SEQ LD 291> was predicted to encode a protein having amino acid sequence <SEQ ID 292> 1 MSVFQTAFFM FQKHLQKASD SWGGTLYW ATPIGNLADI TLRALAVLQK 51 ADIICAEDTR VTAQLLSAYG IQGRLVSVRE HNERQMADKV IGFLSDGLW 101 AQVSDAGTPA VCDPGAKLAR RVREAGFKW PWGASAVMA ALSVAGVAES 151 DFYFNGFVPP KSGERRKLFA KWVRAAFPW MFETPHRIGA TLADMAELFP 201 ERRLMLAREI TKTFETFLSG TVGEIQTALA ADGNQSRGEM VLVLYPAQDE 251 KHEGLSESAQ NAMKILAAEL PTKQAAELAA KITGEGKKAL YDLALSWKNK 301 After further analysis, the following gonococcal DNA sequence <SEQ ID 293> was identified: 1 ATGTTTCAGA AACACTTGCA GAAAGCCTCC GACAGCGTCG TCGGAGGGAC 51 ATTATACGTG GTTGCCACGC CCATCGGCAA TTTGGCAGAC ATTACCCTGC 101 GCGCTTTGGC GGTATTGCAA AAGGCGGACA TCATTTGTGC CGAAGACACG 151 CGCGTTACTG CGCAGCTTTT GAGCGCGTAC GGCATTCAGG GCAGGTTGGT 201 CAGTGTGCGC GAACACAACG AGCGGCAGAT GGCGGACAAG GTAATCGGTT 251 TCCTTTCAGA CGGCCTGGTT GTGGCGCAGG TTTCCGATGC GGGTACGCCG 301 GCCGTGTGCG ACCCGGGCGC GAAACTCGCC CGCCGCGTGC GCGAAGCAGG 351 GTTCAAAGTC GTTCCCGTCG TGGGCGCAAG CGCGGTAATG GCGGCGTTGA 401 GTGTGGCCGG TGTGGCGGAA TCCGATTTTT ATTTCAACGG TTTTGTACCG 451 CCGAAATCGG GCGAACGTAG GAAATTGTTT GCCAAATGGG TGCGGGCGGC 501 ATTTCCTGTC GTCATGTTTG AAACGCCGCA CCGAATCGGG GCAACGCTTG 551 CCGATATGGC GGAATTGTTC CCCGAACGCC GTCTGATGCT GGCGCGCGAA 601 ATCACGAAAA CGTTTGAAAC GTTCTTAAGC GGCACGGTTG GGGAAATTCA 651 GACGGCATTG GCGGCGGACG GCAACCAATC GCGCGGCGAG ATGGTGTTGG 701 TGCTTTATCC GGCGCAGGAT GAAAAACACG AAGGCTTGTC CGAGTCTGCG 751 CAAAATGCGA TGAAAATCCT TGCGGCCGAG CTGCCGACCA AGCAGGCGGC 801 GGAGCTTGCC GCCAAGATTA CAGGTGAGGG CAAAAAGGCT TTGTACGATT 851 TGGCACTGTC GTGGAAAAAC AAATGA This corresponds to the amino acid sequence <SEQ ID 294; ORF75ng-l > • 1 MFQKHLQKAS DS GGTLYV VATPIGNLAD ITLRALAVLQ KADIICAEDT 51 RVTAQLLSAY GIQGRLVSVR EHNERQMADK VIGFLSDGLV VAQVSDAGTP 101 AVCDPGAKLA RRVREAGFKV VPWGASAVM AALSVAGVAE SDFYFNGFVP 151 PKSGERRKLF AKWVRAAFPV VMFETPHRIG ATLADMAELF PERRLMLARE 201 ITKTFETFLS GTVGEIQTAL AADGNQSRGE MVLVLYPAQD EKHEGLSESA 251 QNAMKILAAE LPTKQAAELA AKITGEGKKA LYDLALSWKN K* ORF75ng-l and ORF75-1 show 96.2% identity in 291 aa overlap: 10 20 30 40 50 60 orf 75-1 . pep MFQKHLQKASDSWGGTLYVVATPIGNLADITLRALAVLQKADIICAEDTRVTAQLLSAY I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I orf75ng-l MFQKHLQKASDSWGGTLYWATPIGNLADITLRALAVLQKADIICAEDTRVTAQLLSAY 10 20 30 40 50 60 70 80 90 100 110 120 orf75-1. pep GIQGKLVSVREHNERQMADKIVGYLSDGMWAQVSDAGTPAVCDPGAKLARRVREAGFKV I I IM II I I II I I II II II M: I: I II I M I I II I II II II I II I II I II I I II I I I I I I orf75ng-l GIQGRLVSVREHNERQMADKVIGFLSDGLWAQVSDAGTPAVCDPGAKLARRVREAGFKV 70 80 90 100 110 120 130 140 150 160 170 180 orf75-1. pep VPWGASAVMAALSVAGVEGSDFYFNGFVPPKSGERRKLFAKWVRAAFPIVMFETPHRIG II I II I II II II II II II I I II I I I I I I I I II II I II I I I I II II I I : II II I I I I I I orf75ng-l VPWGASAVMAALSVAGVAESDFYFNGFVPPKSGERRKLFAKWVRAAFPWMFETPHRIG 130 140 150 160 170 180 190 200 210 220 230 240 orf75-1.pep ATLADMAELFPERRLMLAREITKTFETFLSGTVGEIQTALSADGNQSRGEMVLVLYPAQD I I II II I I II II I I I I II II I I II II I I I II II I II II I I : I I I I I I I II II II I I I I II orf75ng-l ATLADMAELFPERRLMLAREITKTFETFLSGTVGEIQTALAADGNQSRGEMVLVLYPAQD 190 200 210 220 230 240 250 260 270 280 290 orf75-1.pep EKHEGLSESAQNIMKILTAELPTKQAAELAAKITGEGKKALYDLALSWKNKX II I I II I II II I II II : II I II I II I II II II I II II I I I I I II II I I II I orf75ng-l EKHEGLSESAQNAMKILAAELPTKQAAELAAKITGEGKKALYDLALSWKNKX 250 260 270 280 290 Furthermore, ORG75ng-l shows significant homology to a hypothetical E.coli protein: sp|P45528 |YRAL_ECOLI HYPOTHETICAL 31.3 KD PROTEIN IN AGAI-MTR INTERGENIC REGION (F286) >gi I 606086 (U18997) ORF_f286 [Escherichia coli] >gi 11789535 (AE000395) hypothetical 31.3 kD protein in agai-mtr intergenic region [Escherichia coli] Length = 286 Score = 218 bits (550), Expect = 3e-56 Identities = 128/284 (45%), Positives = 171/284 (60%), Gaps = 4/284 (1%) Query : 4 KHLQKASDSWGGTLYWATPIGNLADITLRALAVLQKADIICAEDTRVTAQLLSAYGIQ 63 K Q A +S G LY+V TPIGNLADIT RAL VLQ D+I AEDTR T LL +GI Sbj ct : 2 KQHQSADNSQ— GQLYIVPTPIGNLADITQRALEVLQAVDLIAAEDTRHTGLLLQHFGIN 59 Query : 64 GRLVSVREHNERQMADKVIGFLSDGLWAQVSDAGTPAVCDPGAKLARRVREAGFKWPV 123 RL ++ +HNE+Q A+ ++ L +G +A VSDAGTP + DPG L R REAG +WP+ Sbj ct : 60 ARLFALHDHNEQQKAETLLAKLQEGQNIALVSDAGTPLINDPGYHLVRTCREAGIRWPL 119 Query : 124 VGASAVMAALSVAGVAESDFYFNGFVPPKSGERRKLFAKWVRAAFPWMFETPHRIGATL 183 G A + ALS AG+ F + GF+P KS RR ++ +E+ HR+ +L Sbj ct : 120 PGPCAAITALSAAGLPSDRFCYEGFLPAKSKGRRDALKAIEAEPRTLIFYESTHRLLDSL 179 Query : 184 ADMAELFPERR-LMLAREITKTFETFLSGTVGEIQTALAADGNQSRGEMVLVLYPAQDEK 242 D+ + E R ++LARE+TKT+ET VGE+ + D N+ +GEMVL++ + Sbj ct : 180 EDIVAVLGESRYWLARELTKTWETIHGAPVGELLAWVKEDENRRKGEMVLIV-EGHKAQ 238 Query : 243 HEGLSESAQNAMKILAAELPTKQAAELAAKITGEGKKALYDLAL 286 E L A + +L AELP K+AA LAA+I G K ALY AL Sbj ct : 239 EEDLPADALRTLALLQAELPLKKAAALAAEIHGVKKNALYKYAL 282 Based on this analysis, including the presence of a putative transmembrane domain in the gonococcal protein, it is predicted that the proteins from N. meningitidis and N.gonorrhoeae, and their epitopes, could be useful antigens for vaccines or diagnostics, or for raising antibodies. Example 35 The following partial DNA sequence was identified in N. meningitidis <SEQ ID 295>: 1 ATGAAACAGA AAAAAACCGC TGCCGCAGTT ATTGCTGCAA TGTTGGCAGG 51 TTTTGCGGCA GC.AAAGCAC CCGAAATCGA CCCGGCTTTG // 651 GAGTTGG TCAGAAACCA GTTGGAGCAG GGTTTGAGAC 701 AGGAAAAAGC CCGCTTGAAA ATCGATGCCC TTTTGGAAGA AAACGGTGTC 751 AAACCGTAA This corresponds to the amino acid sequence <SEQ ID 296; ORF76>: 1 MKQKKTAAAV IAAMLAGFAA XKAPEIDPAL // 201 ELVRNQLEQG LRQEKARLKI DALLEENGVK 251 P* Further work revealed the complete nucleotide sequence <SEQ LD 297>: 1 ATGAAACAGA AAAAAACCGC TGCCGCAGTT ATTGCTGCAA TGTTGGCAGG 51 TTTTGCGGCA GCCAAAGCAC CCGAAATCGA CCCGGCTTTG GTGGATACGC 101 TGGTGGCGCA GATCATGCAG CAGGCAGACC GGCATGCGGA GCAGTCCCAA 151 AAACCGGACG GGCAGGCAAT CCGAAACGAT GCCGTCCGCC GGCTACAAAC 201 TTTGGAAGTT TTGAAAAACA GGGCATTGAA GGAAGGTTTG GATAAGGATA 251 AGGATGTCCA AAACCGCTTT AAAATCGCCG AAGCGTCTTT TTATGCCGAG 301 GAGTACGTCC GTTTTCTGGA ACGTTCGGAA ACGGTTTCCG AAGACGAGCT 351 GCACAAGTTT TACGAACAGC AAATCCGCAT GATCAAATTG CAGCAGGTCA 401 GCTTCGCAAC CGAAGAGGAG GCGCGTCAGG CGCAGCAGCT CCTGCTCAAA 451 GGGCTGTCTT TTGAAGGGCT GATGAAGCGT TATCCGAACG ACGAGCAGGC 501 TTTTGACGGT TTCATTATGG CGCAGCAGCT TCCCGAGCCG CTGGCTTCGC 551 AGTTTGCCGC GATGAATCGG GGCGACGTTA CCCGCGATCC GGTCAAATTG 601 GGCGAACGCT ATTATCTGTT CAAACTCAGC GAGGTCGGGA AAAACCCCGA 651 CGCGCAGCCT TTCGAGTTGG TCAGAAACCA GTTGGAGCAG GGTTTGAGAC 701 AGGAAAAAGC CCGCTTGAAA ATCGATGCCC TTTTGGAAGA AAACGGTGTC 751 AAACCGTAA This corresponds to the amino acid sequence <SEQ ID 298; ORF76-l>: 1 MKQKKTAAAV IAAMLAGFAA AKAPEIDPAL VDTLVAQIMQ QADRHAEQSQ 51 KPDGQAIRND AVRRLQTLEV LKNRALKEGL DKDKDVQNRF KIAEASFYAE 101 EYVRFLERSE TVSEDELHKF YEQQIRMIKL QQVSFATEEE ARQAQQLLLK 151 GLSFEGLMKR YPNDEQAFDG FIMAQQLPEP LASQFAAMNR GDVTRDPVKL 201 GERYYLFKLS EVGKNPDAQP FELVRNQLEQ GLRQEKARLK IDALLEENGV 251 KP* Computer analysis of this amino acid sequence gave the following results: Homology with a predicted ORF from N.meninsitidis (strain Al ORF76 shows 96.7% identity over a 30aa overlap and 96.8% identity over a 31aa overlap with an ORF (ORF76a) from strain A of N meningitidis: 10 20 30 orf76. pep MKQKKTAAAVIAAMLAGFAAXKAPEIDPAL I I I I I II II I I I I I II I I II I I I I I II II orf76a MKQKKTAAAVIAAMLAGFAAAKAPEIDPALVDTLVAQIMQQADRHAEQSQKPDGOAIRΝD 10 20 30 40 50 60 // 70 80 90 orf76. pep XELVRNQLEQGLRQEKARLKIDALLEENGVKPX II II II I II I I I II II I II I II : II I II II I I orf76a DVTRDPVKLGERYYLFKLSEVGKNPDAQPFELVRNQLEQGLRQEKARLKIDAILEENGVKPX 200 210 220 230 240 250 The complete length ORF76a nucleotide sequence <SEQ ID 299> is: 1 ATGAAACAGA AAAAAACCGC TGCCGCAGTT ATTGCTGCAA TGTTGGCAGG 51 TTTTGCGGCA GCCAAAGCAC CCGAAATCGA CCCGGCTTTG GTGGATACGC 101 TGGTGGCGCA GATCATGCAG CAGGCAGACC GGCATGCGGA GCAGTCCCAA 151 AAACCGGACG GGCAGGCAAT CCGAAACGAT GCCGTCCGTC GGCTGCAAAC 201 TTTGGAAGTT TTGAAAAACA GGGCATTGAA GGAAGGTTTG GATAAGGATA 251 AGGATGTCCA AAACCGCTTT AAAATCGCCG AAGCGTCTTT TTATGCCGAG 301 GAGTACGTCC GTTTTCTGGA ACGTTCGGAA ACGGTTTCCG AAAGCGCACT 351 GCGTCAGTTT TATGAGCGGC AAATCCGCAT GATCAAATTG CAGCAGGTCA 401 GCTTCGCAAC CGAAGAGGAG GCGCGTCAGG CGCAGCAGCT CCTGCTCAAA 451 GGGCTGTCTT TTGAAGGGCT GATGAAGCGT TATCCGAACG ACGAGCAGGC 501 TTTTGACGGT TTCATTATGG CGCAGCAGCT TCCCGAGCCG CTGGCTTCGC 551 AGTTTGCAGC GATGAATCGG GGCGACGTTA CCCGCGATCC GGTCAAATTG 601 GGCGAACGCT ATTATCTGTT CAAACTCAGC GAGGTCGGGA AAAACCCCGA 651 CGCGCAGCCT TTCGAGTTGG TCAGAAACCA GTTGGAACAA GGTTTGAGAC 701 AGGAAAAAGC CCGCTTGAAA ATCGATGCCA TTTTGGAAGA AAACGGTGTC 751 AAACCGTAA This encodes a protein having amino acid sequence <SEQ ID 300>: 1 MKQKKTAAAV IAAMLAGFAA AKAPEIDPAL VDTLVAQIMQ QADRHAEQSQ 51 KPDGQAIRND AVRRLQTLEV LKNRALKEGL DKDKDVQNRF KIAEASFYAE 101 EYVRFLERSE TVSESALRQF YERQIRMIKL QQVSFATEEE ARQAQQLLLK 151 GLSFEGLMKR YPNDEQAFDG FIMAQQLPEP LASQFAAMNR GDVTRDPVKL 201 GERYYLFKLS EVGKNPDAQP FELVRNQLEQ GLRQEKARLK IDAILEENGV 251 KP* ORF76a and ORF76-1 show 97.6% identity in 252 aa overlap: 10 20 30 40 50 60 orf76a. pep MKQKKTAAAVIAAMLAGFAAAKAPEIDPALVDTLVAQIMQQADRHAEQSQKPDGQAIRND II II I I I I I I I II II I I I II I II I II I I II II I II II II II II II II I II I II I II I I I I orf 6-1 MKQKKTAAAVIAAMLAGFAAAKAPEIDPALVDTLVAQIMQQADRHAEQSQKPDGQAIRND 10 20 30 40 50 60 70 80 90 100 110 120 orf76a . pep AVRRLQTLEVLKNRALKEGLDKDKDVQNRFKIAEASFYAEEYVRFLERSETVSESALRQF I I II I II I II II II I I I I II II II I I I I II II II I II II II II II II I II I II I : \ : : \ orf76-1 AVRRLQTLEVLKNRALKEGLDKDKDVQNRFKIAEASFYAEEYVRFLERSETVSEDELHKF 70 80 90 100 110 120 130 140 150 160 170 180 orf76a. pep YERQIRMIKLQQVSFATEEEARQAQQLLLKGLSFEGLMKRYPNDEQAFDGFIMAQQLPEP I I: I II II I II II I I I II I I II I I I I I I II II II I II II I II I II II I II I II II I I I I I orf76-1 YEQQIRMIKLQQVSFATEEEARQAQQLLLKGLSFEGLMKRYPNDEQAFDGFIMAQQLPEP 130 140 150 160 170 180 190 200 210 220 230 240 orf76a . pep LASQFAAMNRGDVTRDPVKLGERYYLFKLSEVGKNPDAQPFELVRNQLEQGLRQEKARLK I I II I I I I II I II II II II II I I II II II II I II II II II I I II I I I I II I II II II I I I orf76-1 LASQFAAMNRGDVTRDPVKLGERYYLFKLSEVGKNPDAQPFELVRNQLEQGLRQEKARLK 190 200 210 220 230 240 250 orf76a. pep IDAILEENGVKPX I I I: I I I I II I I I orf76-1 IDALLEENGVKPX 250 Homology with a predicted ORF from N.sonorrhoeae The aligned aa sequences of ORF76 and a predicted ORF (ORF76.ng) from N. gonorrhoeae of the Ν- and C-termini show 96.7 % and 100% identity in 30 and 31 overlap, respectively: orf76. pep MKQKKTAAAVIAAMLAGFAAXKAPEIDPAL 30 II I I I II I I I II II II II II II I I I I I II orf76ng MKQKKTAAAVIAAMLAGFAAAKAPEIDPALVDTLVAQIMQQADRHAEQSQRPDGQAIRND 60 // orf 6.pep ELVRNQLEQGLRQEKARLKIDALLEENGVKP 251 II II I II II II I II I I I I I I I I II II I I I II orf 6ng VTRNPVKLGERYYLFKLGAVGKNPDAQPFELVRNQLEQGLRQEKARLKIDALLEENGVKP 251 The complete length ORF76ng nucleotide sequence <SEQ ID 301 > is: 1 ATGAAACAGA AAAAGACCGC TGCCGCAGTT ATTGCTGCAA TGTTGGCAGG 51 TTTTGCGGCA GCCAAAGCAC CCGAAATCGA CCCGGCTTTG GTGGATACGC 101 TGGTGGCGCA GATCATGCAG CAGGCAGACC GGCATGCGGA GCAGTCCCAA 151 AGACCGGACG GGCAGGCAAT CCGAAACGAT GCCGTCCGCC GGCTGCAAAC 201 TTTGGAAGTT TTGAAAAACA GGGCATTGAA GGAAGGTTTG GATAAGGATA 251 AGGATGTCCA AAACCGCTTT AAAATCGCCG AAGCGTCTTT TTATGCCGAG 301 GAGTACGTCC GTTTTCTGGA ACGTTCGGAA ACGGTTTCCG AAAGCGCACT 351 GCGTCAGTTT TATGAGCGGC AAATCCGCAT GATCAAATTG CAGCAGGTCA 401 GCTTCGCAAC CGAAGAGGAG GCGCGTCAGG CGCAGCAGCT CCTGCTCAAA 451 GGGCTGTCTT TTGAAGGGCT GATGAAGCGT TATCCGAACG ACGAGCAGGC 501 GTTCGACGGT TTCATTATGG CGCAGCAGCT TCCCGAGCCG CTGGCTTcgc 551 agtttgCCGG TATGAACCGT GGCGACGTTA CCCGCAATCC GGTCAAATTG 601 GGCGAACGCT ATTACCTGTT CAAACTCGGC GCGGTCGGGA AAAACCCCGA 651 CGCGCAGCCT TTCGAGTTGG TCAGAAACCA GTTGGAACAA GGTTTGAGGC 701 AGGAAAAAGC CCGCTTGAAA ATCGATGCCC TTTTGGAaga Aaacggtgtc 751 AaacCGTAA This encodes a protein having amino acid sequence <SEQ ID 302>: 1 MKQKKTAAAV IAAMLAGFAA AKAPEIDPAL VDTLVAQIMQ QADRHAEQSQ 51 RPDGQAIRND AVRRLQTLEV LKNRALKEGL DKDKDVQNRF KIAEASFYAE 101 EYVRFLERSE TVSESALRQF YERQIRMIKL QQVSFATEEE ARQAQQLLLK 151 GLSFEGLMKR YPNDEQAFDG FIMAQQLPEP LASQFAGMNR GDVTRNPVKL 201 GERYYLFKLG AVGKNPDAQP FELVRNQLEQ GLRQEKARLK IDALLEENGV 251 KP* ORF76ng and ORF76-1 show 96.0% identity in 252 aa overlap 10 20 30 40 50 60 orf76-1. pep MKQKKTAAAVIAAMLAGFAAAKAPEIDPALVDTLVAQIMQQADRHAEQSQKPDGQAIRND II I I II II II II II I I I I II I I II I II II I II I II I I I I II II I II II I I: II I I I II II orf76ng MKQKKTAAAVIAAMLAGFAAAKAPEIDPALVDTLVAQIMQQADRHAEQSQRPDGQAIRND 10 20 30 40 50 60 70 80 90 100 110 120 orf76-1. pep AVRRLQTLEVLKNRALKEGLDKDKDVQNRFKIAEASFYAEEYVRFLERSETVSEDELHKF II I II I I I I II I I I I I I II I I I I I II I II I II I II II II II II I I I II I II II I : MM orf76ng AVRRLQTLEVLKNRALKEGLDKDKDVQNRFKIAEASFYAEEYVRFLERSETVSESALRQF 70 80 90 100 110 120 130 140 150 160 170 180 orf76-1. pep YEQQIRMIKLQQVSFATEEEARQAQQLLLKGLSFEGLMKRYPNDEQAFDGFIMAQQLPEP I I: I I I I I I I I I II I II II I I II I II I II I II II I II II II II I I I II I II II II I I I I I orf76ng YERQIRMIKLQQVSFATEEEARQAQQLLLKGLSFEGLMKRYPNDEQAFDGFIMAQQLPEP 130 140 150 160 170 180 190 200 210 220 230 240 orf76-1. pep LASQFAAMNRGDVTRDPVKLGERYYLFKLSEVGKNPDAQPFELVRNQLEQGLRQEKARLK orf76ng LASQFAGMNRGDVTRNPVKLGERYYLFKLGAVGKNPDAQPFELVRNQLEQGLRQEKARLK 190 200 210 220 230 240 250 orf76-1. pep IDALLEENGVKPX I I I I I I I I I I I I I orf76ng IDALLEENGVKPX 250 Furthermore, ORF76ng shows significant homology to a B.subtilis export protein precursor: sp|P24327 |PRSA_BACSU PROTEIN EXPORT PROTEIN PRSA PRECURSOR >gi | 98227 | pir || S15269 33K lipoprotein - Bacillus subtilis >gi 139782 (X57271) 33kDa lipoprotein [Bacillus subtilis] >gi|2226124 |gnl| PID|e325181 (Y14077) 33kDa lipoprotein [Bacillus subtilis] >gi I 26333311 gnl I PID|ell82997 (Z99109) molecular chaperonin [Bacillus subtilis] Length = 292 Score = 50.4 bits (118), Expect = le-05 Identities = 48/199 (24%), Positives = 82/199 (41%), Gaps = 32/199 (16%) Query: 70 VLKNRALKEGLDK DKDVQNRFKIAEASF YAEEYVRFLERSETVSE 114 VL ++ LDK DK++ N+ K + Y ++Y++ + E +++ Sbjct: 53 VLTQLVQEKVLDKKYKVSDKEIDNKLKEYKTQLGDQYTALEKQYGKDYLKEQVKYELLTQ 112 Query: 115 SA LRQFYERQIRMIKLQQVSFATEEEARQAQQLLLKGLSFEGLMKRYPN 163 A +++++E 1+ + A ++ A + ++ L KG FE L K Y Sbjct: 113 KAAKDNIKVTDADIKEYWEGLKGKIRASHILVADKKTAEEVEKKLKKGEKFEDLAKEYST 172 Query: 164 DEQAFDG FIMAQQLPEPLASQFAAMNRGDVTRDPVKLGERYYLFKLSEVGKNPDA 218 D A G F Q+ E + + G+V+ DPVK Y++ K +E D Sbjct: 173 DSSASKGGDLGWFAKEGQMDETFSKAAFKLKTGEVS-DPVKTQYGYHIIKKTEERGKYDD 231 Query: 219 QPFELVRNQLEQGLRQEKA 237 EL LEQ L A Sbjct: 232 MKKELKSEVLEQKLNDNAA 250 Based on this analysis, including the presence of a putative leader sequence and a RGD motif in the gonococcal protein, it was predicted that the proteins from N meningitidis and N.gonorrhoeae, and their epitopes, could be useful antigens for vaccines or diagnostics, or for raising antibodies. ORF76-1 (27.8kDa) was cloned in the pET vector and expressed in E.coli, as described above. The products of protein expression and purification were analyzed by SDS-PAGE. Figure 10A shows the results of affinity purification of the His-fusion protein, Purified His-fusion protein was used to immunise mice, whose sera were used for Western blot (Figure 10B), ELISA (positive result), and FACS analysis (Figure 10C). These experiments confirm that ORF76-1 is a surface-exposed protein, and that it is a useful immunogen. Example 36 The following partial DΝA sequence was identified in N. meningitidis <SEQ ID 303>: 1 ATGAAAAAAT CTTTCCTTAC GCTTGTTCTG TATTCGTCTT TACTTACCGC 51 CAGCGAAATT GCCΪTACCCC TTGGAATTGG GGATTGAAAC CTTACCGGCG 101 GCAAAAATTG CGGAAACGTT TGCGCTGACA TTTGTGATTG CTGCGCTGTA 151 TCTGTTTGCG CGTAATAAGG TGACGCGTTT GTTGATTGCG GTGTTTTTTG 201 CGTTCAGCAT TATTGCCAAC AATGTGCATT ACGCGGATTA TCAAAGCTGG 251 ATGACG // 1201 CAAACCGTAT TCGAGCAGCT GCAAAAGACT CCTGACGGCA 1251 ACTGGCTGTT TGCCTATACC TCCGATCATG GCCAGTATGT TCGCCAAGAT 1301 ATCTACAATC AAGGCACGGT GCAGCCCGAC AGCTATCTCG TGCCGCTAGT 1351 GTTGTACAGC CCGGATAAGG CCGTGCAACA GGCTGCCAAC CAGGCTTTTG 1401 CGCCTTGCGA GATTGCCTTC CATCAGCAGC TTTCAACGTT CCTGATTCAC 1451 ACGTTGGGCT ACGATATGCC GGTTTCAGGT TGTCGCGAAG GCTCGGTAAC 1501 GGGCAACCTG ATTACGGGTG ATGCAGGCAG CTTGAACATT CGCGACGGCA 1551 AGGCGGAATA TGTTTATCCG CAATGA This corresponds to the amino acid sequence <SEQ ID 304; ORF81>: 1 MKKΞFLTLVL YΞSLLTASEI AYPLELGIET LPAAKIAETF ALTFVIAALY 51 LFARNKVTRL LIAVFFAFSI IANNVHYADY QSWMT // 401 ...QTVFEQL QKTPDGNWLF AYTSDHGQYV RQDIYNQGTV QPDSYLVPLV 451 LYSPDKAVQQ AANQAFAPCE IAFHQQLSTF LIHTLGYDMP VSGCREGSVT 501 GNLITGDAGS LNIRDGKAEY VYPQ* Further work revealed the complete nucleotide sequence <SEQ ID 305>: 1 ATGAAAAAAT CTTTCCTTAC GCTTGTTCTG TATTCGTCTT TACTTACCGC 51 CAGCGAAATT GCCTATCGCT TTGTATTTGG GATTGAAACC TTACCGGCGG 101 CAAAAATTGC GGAAACGTTT GCGCTGACAT TTGTGATTGC TGCGCTGTAT 151 CTGTTTGCGC GTTATAAGGT GACGCGTTTG TTGATTGCGG TGTTTTTTGC 201 GTTCAGCATT ATTGCCAACA ATGTGCATTA CGCGGTTTAT CAAAGCTGGA 251 TGACGGGCAT CAATTATTGG CTGATGCTGA AAGAGGTTAC CGAAGTCGGC 301 AGCGCGGGTG CGTCGATGTT GGATAAGTTG TGGCTGCCTG TGTTGTGGGG 351 CGTGTTGGAA GTCATGTTGT TTTGCAGCCT TGCCAAGTTC CGCCGTAAGA 401 CGCATTTTTC TGCCGATATA CTGTTTGCCT TCCTAATGCT GATGATTTTC 451 GTGCGTTCGT TCGACACGAA ACAAGAGCAC GGTATTTCGC CCAAACCGAC 501 ATACAGCCGC ATCAAAGCCA ATTATTTCAG CTTCGGTTAT TTTGTCGGAC 551 GCGTGTTGCC GTATCAGTTG TTTGATTTAA GCAGGATTCC CGCCTTTAAG 601 CAGCCTGCTC CAAGCAAAAT CGGGCAGGGC AGTGTTCAAA ATATCGTCCT 651 GATTATGGGC GAAAGCGAAA GCGCGGCGCA TTTGAAGCTG TTTGGCTACG 701 GACGCGAAAC TTCGCCGTTT TTAACCCGGC TGTCGCAAGC CGATTTTAAG 751 CCGATTGTGA AACAAAGTTA TTCCGCAGGC TTTATGACTG CAGTGTCCCT 801 GCCCAGTTTT TTCAATGCGA TACCGCACGC CAACGGCTTG GAACAAATCA 851 GCGGCGGCGA TACCAATATG TTCCGCCTCG CCAAAGAGCA GGGCTATGAA 901 ACGTATTTTT ACAGCGCGCA GGCGGAAAAC GAGATGGCGA TTTTGAACTT 951 AATCGGTAAG AAATGGATAG ACCATCTGAT TCAGCCGACG CAACTTGGCT 1001 ACGGCAACGG CGACAATATG CCCGATGAGA AGCTGCTGCC GTTGTTCGAC 1051 AAAATCAATT TGCAGCAGGG CAAGCATTTT ATCGTGTTGC ACCAACGCGG 1101 TTCGCACGCC CCATACGGCG CATTGTTGCA GCCTCAAGAT AAAGTATTCG 1151 GCGAAGCCGA TATTGTGGAT AAGTACGACA ACACCATCCA CAAAACCGAC 1201 CAAATGATTC AAACCGTATT CGAGCAGCTG CAAAAGCAGC CTGACGGCAA 1251 CTGGCTGTTT GCCTATACCT CCGATCATGG CCAGTATGTT CGCCAAGATA 1301 TCTACAATCA AGGCACGGTG CAGCCCGACA GCTATCTCGT GCCGCTAGTG 1351 TTGTACAGCC CGGATAAGGC CGTGCAACAG GCTGCCAACC AGGCTTTTGC 1401 GCCTTGCGAG ATTGCCTTCC ATCAGCAGCT TTCAACGTTC CTGATTCACA 1451 CGTTGGGCTA CGATATGCCG GTTTCAGGTT GTCGCGAAGG CTCGGTAACG 1501 GGCAACCTGA TTACGGGTGA TGCAGGCAGC TTGAACATTC GCGACGGCAA 1551 GGCGGAATAT GTTTATCCGC AATGA This corresponds to the .amino acid sequence <SEQ ID 306; ORF81-l>: 1 MKKSFLTLVL YSSLLTASEI AYRFVFGIET LPAAKIAETF ALTFVIAALY 51 LFARYKVTRL LIAVFFAFSI IANNVHYAVY QSWMTGINYW LMLKEVTEVG 101 SAGASMLDKL WLPVLWGVLE VMLFCSLAKF RRKTHFSADI LFAFLMLMIF 151 VRSFDTKQEH GISPKPTYSR IKANYFSFGY FVGRVLPYQL FDLSRIPAFK 201 QPAPSKIGQG SVQNIVLIMG ESESAAHLKL FGYGRETSPF LTRLSQADFK 251 PIVKQSYSAG FMTAVSLPSF FNAIPHANGL EQISGGDTNM FRLAKEQGYE 301 TYFYSAQAEN EMAILNLIGK KWIDHLIQPT QLGYGNGDNM PDEKLLPLFD 351 KINLQQGKHF IVLHQRGSHA PYGALLQPQD KVFGEADIVD KYDNTIHKTD 401 QMIQTVFEQL QKQPDGNWLF AYTSDHGQYV RQDIYNQGTV QPDSYLVPLV 451 LYSPDKAVQQ AANQAFAPCE IAFHQQLSTF LIHTLGYDMP VSGCREGSVT ' 501 GNLITGDAGS LNIRDGKAEY VYPQ* Computer analysis of this amino acid sequence gave the following results: Homology with a predicted ORF from N. meningitidis (strain Al ORF81 shows 84.7% identity over a 85aa overlap and 99.2% identity over a 121aa overlap with an ORF (ORF81a) from strain A of N. meningitidis: 10 20 30 40 50 60 orf81.pep MKKSFLTLVLYSSLLTASEIAYPLELGIETLPAAKIAETFALTFVIAALYLFARNKVTRL I II I::: I I I II I I I I I I II I : M II I I I II I M II I I I I II II I II I II I Mill orf8la MKKSLFVLFLYSSLLTASEIAYRFVFGIETLPAAKMAETFALTFVIAALYLFARYKATRL 10 20 30 40 50 60 70 80 orf81. pep LIAVFFAFSIIANNVHYADYQSWMT I II I II II II I I II II I I MMM orf8la LIAVFFAFSIIANNVHYAVYQSWITGINYWLMLKEITEVGGAGASMLDKLWLPALWGVLE 70 80 90 100 110 120 // 120 130 140 orf81.pep QTVFEQLQKTPDGNWLFAYTSDHGQYVRQD II I II I II I I II I I I I I I I I II II I I II I orf81a IPHANGLEQISGGDIVDKYDNTIHKTDQMIQTVFEQLQKQPDGNWLFAYTSDHGQYVRQD 280 290 300 310 320 330 150 160 170 180 190 200 orf81. pep IYNQGTVQPDSYLVPLVLYSPDKAVQQAANQAFAPCEIAFHQQLSTFLIHTLGYDMPVSG I I II I I I II II II II I I I II I I II I M I II I II II I II II I II II II I I I I II I II I I I I orf8la IYNQGTVQPDSYLVPLVLYSPDKAVQQAANQAFAPCEIAFHQQLSTFLIHTLGYDMPVSG 340 350 360 370 380 390 210 220 230 orf81. pep CREGSVTGNLITGDAGSLNIRDGKAEYVYPQX II I II II II I I I II II I II I II I I I II I II II orf81a CREGSVTGNLITGDAGSLNIRDGKAEYVYPQX 400 410 420 The complete length ORF8 la nucleotide sequence <SEQ ID 307> is: 1 ATGAAAAAAT CCCTTTTCGT TCTCTTTCTG TATTCGTCCC TACTTACTGC 51 CAGCGAAATT GCTTATCGCT TTGTATTCGG AATTGAAACC TTACCGGCTG 101 CAAAAATGGC AGAAACGTTT GCGCTGACAT TTGTGATTGC TGCGCTGTAT 151 CTGTTTGCGC GTTATAAGGC AACGCGTTTG TTGATTGCGG TGTTTTTCGC 201 GTTCAGCATT ATTGCCAACA ATGTGCATTA CGCGGTTTAT CAAAGCTGGA 251 TAACGGGCAT TAATTATTGG CTGATGCTGA AAGAGATTAC CGAAGTTGGC 301 GGCGCAGGGG CGTCGATGTT GGATAAGTTG TGGCTGCCTG CGTTGTGGGG 351 CGTGTTGGAA GTCATGTTGT TTTGCAGCCT TGCCAAGTTC CGCCGTAAGA 401 CGCATTTTTC TGCCGATATA CTGTTTGCCT TCCTAATGCT GATGATTTTC 451 GTGCGTTCGT TCGACACGAA ACAAGAACAC GGTATTTCGC CCAAACCGAC 501 ATACAGCCGC ATCAAAGCCA ATTATTTCAG CTTCGGTTAT TTTGTCGGAC 551 GCGTGTTGCC GTATCAGTTG TTTGATTTAA GCAAGATTCC TGTGTTCAAA 601 CAGCCTGCTC CAAGCAGAAT CGGGCAAGGC AGTATTCAAA ATATCGTCCT 651 GATTATGGGC GAAAGCGAAA GCGCGGCGCA TTTGAAATTG TTTGGCTACG 701 GGCGCGAAAC TTCGCCGTTT TTGACCCAGC TTTCGCAAGC CGATTTTAAG 751 CCGATTGTGA AACAAAGTTA TTCCGCAGGC TTTATGACGG CAGTATCCCT 801 GCCCAGTTTC TTTAACGTCA TACCGCATGC CAACGGCTTG GAACAAATCA 851 GCGGCGGCGA TATTGTGGAT AAGTACGACA ACACCATCCA CAAAACCGAC 901 CAAATGATTC AAACCGTATT CGAGCAGCTG CAAAAGCAGC CTGACGGCAA 951 CTGGCTGTTT GCCTATACCT CCGATCATGG CCAGTATGTT CGCCAAGATA 1001 TCTACAATCA AGGCACGGTG CAGCCCGACA GCTATCTCGT GCCGCTGGTG 1051 TTGTACAGCC CGGATAAGGC CGTGCAACAG GCTGCCAACC AGGCTTTTGC 1101 GCCTTGCGAG ATTGCCTTCC ATCAGCAGCT TTCAACGTTC CTGATTCACA 1151 CGTTGGGCTA CGATATGCCG GTTTCAGGTT GTCGCGAAGG CTCGGTAACG 1201 GGCAACCTGA TTACGGGTGA TGCAGGCAGC TTGAACATTC GCGACGGCAA 1251 GGCGGAATAT GTTTATCCGC AATGA This encodes a protein having amino acid sequence <SEQ ID 308>: 1 MKKSLFVLFL YSSLLTASEI AYRFVFGIET LPAAKMAETF ALTFVIAALY 51 LFARYKATRL LIAVFFAFSI IANNVHYAVY QSWITGINYW LMLKEITEVG 101 GAGASMLDKL WLPALWGVLE VMLFCSLAKF RRKTHFSADI LFAFLMLMIF 151 VRSFDTKQEH GISPKPTYSR IKANYFSFGY FVGRVLPYQL FDLSKIPVFK 201 QPAPSRIGQG SIQNIVLIMG ESESAAHLKL FGYGRETSPF LTQLSQADFK 251 PIVKQSYSAG FMTAVSLPSF FNVIPHANGL EQISGGDIVD KYDNTIHKTD 301 QMIQTVFEQL QKQPDGNWLF AYTSDHGQYV RQDIYNQGTV QPDSYLVPLV 351 LYSPDKAVQQ AANQAFAPCE IAFHQQLSTF LIHTLGYDMP VSGCREGSVT 401 GNLITGDAGS LNIRDGKAEY VYPQ* ORFδla and ORF81-1 show 77.9% identity in 524 aa overlap: 10 20 30 40 50 60 orf8la. pep MKKSLFVLFLYSSLLTASEIAYRFVFGIETLPAAKMAETFALTFVIAALYLFARYKATRL I I II : : M M II I I II I II II I II II II I II I I M II II II I I II I I I II II I I I : I II orf81-1 MKKSFLTLVLYSSLLTASEIAYRFVFGIETLPAAKIAETFALTFVIAALYLFARYKVTRL 10 20 30 40 50 60 70 80 90 100 110 120 orf81a . pep LIAVFFAFSIIANNVHYAVYQSWITGINYWLMLKEITEVGGAGASMLDKLWLPALWGVLE II II I II I II II I I I II I II I I I : II II I I I I II I : I I I I : II I I II I I II I I : II II I I orf81-1 LIAVFFAFSIIANNVHYAVYQSWMTGINYWLMLKEVTEVGSAGASMLDKLWLPVLWGVLE 70 80 90 100 110 120 130 140 150 160 170 180 orf81a. pep VMLFCSLAKFRRKTHFSADILFAFLMLMIFVRSFDTKQEHGISPKPTYSRIKANYFSFGY II I II II I II II I I II I II I II I II II II II II I I I II I I I I II II II II II II I I I I II orf81-1 VMLFCSLAKFRRKTHFSADILFAFLMLMIFVRΞFDTKQEHGISPKPTYSRIKANYFSFGY 130 140 150 160 170 180 190 200 210 220 230 240 orf81a. pep FVGRVLPYQLFDLSKIPVFKQPAPSRIGQGSIQNIVLIMGESESAAHLKLFGYGRETSPF II I II II I I II I II : I I : I I I II II : II II I : II II I II II I I II I II I II I I II II II I orf81-1 FVGRVLPYQLFDLSRIPAFKQPAPSKIGQGSVQNIVLIMGESESAAHLKLFGYGRETSPF 190 200 210 220 230 240 250 260 270 280 orf81a. pep LTQLSQADFKPIVKQSYSAGFMTAVSLPSFFNVIPHANGLEQISGGD I I : II I II I II II II II I II I I II I II I I II I : II I II I I I I II I I I orf81-1 LTRLSQADFKPIVKQSYSAGFMTAVSLPSFFNAIPHANGLEQISGGDTNMFRLAKEQGYE 250 260 270 280 290 300 orf81a. pep orf81-1 TYFYSAQAENEMAILNLIGKKWIDHLIQPTQLGYGNGDNMPDEKLLPLFDKINLQQGKHF 310 320 330 340 350 360 290 300 310 320 orf81a. pep IVDKYDNTIHKTDQMIQTVFEQLQKQPDGNWLF I II I I I II II I II II I II I II I I II II I I I I I I orf81-1 IVLHQRGSHAPYGALLQPQDKVFGEADIVDKYDNTIHKTDQMIQTVFEQLQKQPDGNWLF 370 380 390 400 410 420 330 340 350 360 370 380 orf8la. pep AYTSDHGQYVRQDIYNQGTVQPDSYLVPLVLYSPDKAVQQAANQAFAPCEIAFHQQLSTF II I II II II M I II I II II I I II I II II I I I I I II II I II I I I II I II I I II II II I I I I orf81-1 AYTSDHGQYVRQDIYNQGTVQPDSYLVPLVLYSPDKAVQQAANQAFAPCEIAFHQQLSTF 430 440 450 460 470 480 390 400 410 420 orf81a.pep LIHTLGYDMPVSGCREGSVTGNLITGDAGSLNIRDGKAEYVYPQX I I I I I II I II II II I I I I II I II II I II II I I I II II I II I II II orf81-1 LIHTLGYDMPVSGCREGSVTGNLITGDAGSLNIRDGKAEYVYPQX 490 500 510 520 Homology with a predicted ORF from N.gonorrhoeae The aligned aa sequences of ORF81 and a predicted ORF (ORFδl.ng) from N gonorrhoeae of the Ν- and C-termini show 82.4 % and 97.5% identity in 85 and 121 overlap, respectively: orf81.pep MKKSFLTLVLYSSLLTASEIAYPLELGIETLPAAKIAETFALTFVIAALYLFARΝKVTRL 60 I II I::: I I II I II II II II I : : I I I II II II : II I I I I II : I I I II I II I I:: II orfβlng MKKSLFVLFLYΞSLLTASEIAYRFVFGIETLPAAKMAETFALTFMIAALYLFARYKASRL 60 orf81. pep LIAVFFAFSIIAΝΝVHYADYQSWMT 85 I I II I II I I: I II I II I I I II I II orfδlng LIAVFFAFSMIAΝΝVHYAVYQSWMTGIΝYWLMLKEVTEVGSAGASMLDKLWLPALWGVAE 120 // orf81. pep QTVFEQLQKTPDGΝWLFAYTSDHGQYVRQD 433 I II I II I II I I I I I II I I I I I II I I I I I I orfδlng ALLQPQDKVFGEADIVDKYDΝTIHKTDQMIQTVFEQLQKQPDGΝWLFAYTSDHGQYVRQD 433 orf81. pep IYΝQGTVQPDSYLVPLVLYSPDKAVQQAAΝQAFAPCEIAFHQQLSTFLIHTLGYDMPVSG 493 I I II I I I I I II l II I I II II I II I M M M I II II I I I I II II I II I II I I I I I I I I I orfδlng IYΝQGTVQPDSYIVPLVLYSPDKAVQQAAΝQAFAPCEIAFHQQLSTFLIHTLGYDMPVSG 493 orf 81 . pep CREGSVTGNLITGDAGSLNIRDGKAEYVYPQ 524 I I I I I I I I I I I I I I I I I I I M M I I I I I I I I orf 81ng CREGSVTGNLITGDAGSLNIRNGKAEYVYPQ 524 The complete length ORF81ng nucleotide sequence <SEQ ID 309> is: 1 ATGAAAAAAT CCCTTTTCGT TCTCTTTCTG TATTCATCCC TACTTACCGC 51 CAGCGAAATC GCCTATCGCT TTGTATTCGG AATTGAAACC TTACCGGCTG 101 CAAAAATGGC GGAAACGTTT GCGCTGACAT TTATGATTGC TGCGCTGTAT 151 CTGTTTGCGC GTTATAAGGC TTCGCGGCTG CTGATTGCGG TGTTTTTCGC 201 GTTCAGCATG ATTGCCAACA ATGTGCATTA CGCGGTTTAT CAAAGCTGGA 251 TGACGGGTAT TAACTATTGG CTGATGCTGA AAGAGGTTAC CGAAGTCGGC 301 AGCGCGGGCG CGTCGATGTT GGATAAGTTG TGGCTGCCTG CTTTGTGGGG 351 CGTGGCGGAA GTCATGTTGT TTTGCAGCCT TGCCAAGTTC CGCCGTAAGA 401 CGCATTTTTC TGCCGATATA CTGTTTGCCT TCCTAATGCT GATGATTTTC 451 GTGCGTTCGT TCGACACGAA ACAAGAGCAC GGTATTTCGC CCAAACCGAC 501 ATACAGCCGC ATCAAAGCCA ATTATTTCAG CTTCGGTTAT TTTGTCGGGC 551 GCGTGTTGCC GTATCAGTTG TTTGATTTAA GCAAGATCCC TGTGTTCAAA 601 CAGCCTGCTC CAAGCAAAAT CGGGCAAGGC AGTATTCAAA ATATCGTCCT 651 GATTATGGGC GAAAGCGAAA GCGCGGCGCA TTTGAAATTG TTTGGTTACG 701 GGCGCGAAAC TTCGCCGTTT TTAACCCGGC TGTCGCAAGC CGATTTTAAG 751 CCGATTGTGA AACAAAGTTA TTCCGCAGGC TTTATGACGG CAGTATCCCT 801 GCCCAGTTTC TTTAACGTCA TACCGCACGC CAACGGCTTG GAACAAATCA 851 GCGGCGGCGA TACCAATATG TTCCGCCTCG CCAAAGAGCA GGGCTATGAA 901 ACGTATTTTT ACAGTGCCCA GGCTGAAAAC CAAATGGCAA TTTTGAACTT 951 AATCGGTAAG AAATGGATAG ACCATCTGAT TCAGCCGACG CAACTTGGCT 1001 ACGGCAACGG CGACAATATG CCCGATGAGA AGCTGCTGCC GTTGTTCGAC 1051 AAAATCAATT TGCAGCAGGG CAGGCATTTT ATCGTGTTGC ACCAACGCGG 1101 TTCGCACGCC CCATACGGCG CATTGTTGCA GCCTCAAGAT AAAGTATTCG 1151 GCGAAGCCGA TATTGTGGAT AAGTACGACA ACACCATCCA CAAAACCGAC 1201 CAAATGATTC AAACCGTATT CGAGCAGCTG CAAAAGCAGC CTGACGGCAA 1251 CTGGCTGTTT GCCTATACCT CCGATCATGG CCAGTATGTG CGCCAAGATA 1301 TCTACAATCA AGGCACGGTG CAGCCCGACA GCTATATTGT GCCTCTGGTT 1351 TTGTACAGCC CGGATAAGGC CGTGCAACAG GCTGCCAACC AGGCTTTTGC 1401 GCCTTGCGAG ATTGCCTTCC ATCAGCAGCT TTCAACGTTC CTGATTCACA 1451 CGTTGGGCTA CGATATGCCG GTTTCAGGTT GTCGCGAAGG CTCGGTAACA 1501 GGCAACCTGA TTACGGGCGA TGCAGGCAGC TTGAACATTC GCAACGGCAA 1551 GGCGGAATAT GTTTATCCGC AATAA This encodes a protein having amino acid sequence <SEQ ID 310>: 1 MKKSLFVLFL YSSLLTASEI AYRFVFGIET LPAAKMAETF ALTFMIAALY 51 LFARYKASRL LIAVFFAFSM IANNVHYAVY QSWMTGINYW LMLKEVTEVG 101 SAGASMLDKL WLPALWGVAE VMLFCSLAKF RRKTHFSADI LFAFLMLMIF 151 VRSFDTKQEH GISPKPTYSR IKANYFSFGY FVGRVLPYQL FDLSKIPVFK 201 QPAPSKIGQG SIQNIVLIMG ESESAAHLKL FGYGRETSPF LTRLSQADFK 251 PIVKQSYSAG FMTAVSLPSF FNVIPHANGL EQISGGDTNM FRLAKEQGYE 301 TYFYSAQAEN QMAILNLIGK KWIDHLIQPT QLGYGNGDNM PDEKLLPLFD 351 KINLQQGRHF IVLHQRGSHA PYGALLQPQD KVFGEADIVD KYDNTIHKTD 401 QMIQTVFEQL QKQPDGNWLF AYTSDHGQYV RQDIYNQGTV QPDSYIVPLV 451 LYSPDKAVQQ AANQAFAPCE IAFHQQLSTF LIHTLGYDMP VSGCREGSVT 501 GNLITGDAGS LNIRNGKAEY VYPQ* ORF81ng and ORF81-1 show 96.4% identity in 524 aa overlap: 10 20 30 40 50 60 orf 81ng-l . pep MKKSLFVLFLYSSLLTASEIAYRFVFGIETLPAAKMAETFALTFMIAALYLFARYKASRL I I I I : : : I I I I I I I I I I I I I I I I I I I I I I I I I I I : I I I I I I I I : I I I I I I I I I I I : : I I orf 81-l MKKSFLTLVLYSSLLTASEIAYRFVFGIETLPAAKIAETFALTFVIAALYLFARYKVTRL 10 20 30 40 50 60 70 80 90 100 110 120 orf 81ng- l . pep LIAVFFAFSMIANNVHYAVYQSWMTGINYWLMLKEVTEVGSAGASMLDKLWLPALWGVAE II I I II I II : I I I I II II I I I I II I II II I II I II II I I I II I I I I I I I I II I : I II I I orf81-l LIAVFFAFSIIANNVHYAVYQSWMTGINYWLMLKEVTEVGSAGASMLDKLWLPVLWGVLE 70 80 90 100 110 120 130 140 150 160 170 180 orfδlng-l.pep VMLFCSLAKFRRKTHFSADILFAFLMLMIFVRSFDTKQEHGISPKPTYSRIKANYFSFGY II I I II I II I I II I II I II I I I I I I I II II II I I I I I II II I II II I II I I II II I I II I orf81-l VMLFCSLAKFRRKTHFSADILFAFLMLMIFVRSFDTKQEHGISPKPTYSRIKANYFSFGY 130 140 150 160 170 180 190 200 210 220 230 240 orf81ng-l . pep FVGRVLPYQLFDLSKIPVFKQPAPSKIGQGSIQNIVLIMGESESAAHLKLFGYGRETSPF I II I I II II II II I: I I : II I M I I M I I I I : I I II II I I I I II II II I II I II II II I I orf81-1 FVGRVLPYQLFDLSRIPAFKQPAPSKIGQGSVQNIVLIMGESESAAHLKLFGYGRETSPF 190 200 210 220 230 240 250 260 270 280 290 300 orf81ng-l .pep LTRLSQADFKPIVKQSYSAGFMTAVSLPSFFNVIPHANGLEQISGGDTNMFRLAKEQGYE I I II II II I II I I I I I II II I II I II I I I II I: II I II I I I I I II I II I I II I I I I I II I orf81-1 LTRLSQADFKPIVKQSYSAGFMTAVSLPSFFNAIPHANGLEQISGGDTNMFRLAKEQGYE 250 260 270 280 290 300 310 320 330 340 350 360 orf81ng-l . pep TYFYSAQAENQMAILNLIGKKWIDHLIQPTQLGYGNGDNMPDEKLLPLFDKINLQQGRHF I II I I I II IM I II I I I I I I I II I II I II II I I II I I II I II I II II I II II I I I I I: II orf81-1 TYFYSAQAENEMAILNLIGKKWIDHLIQPTQLGYGNGDNMPDEKLLPLFDKINLQQGKHF 310 320 330 340 350 360 370 380 390 400 410 420 orf81ng-l.pep IVLHQRGSHAPYGALLQPQDKVFGEADIVDKYDNTIHKTDQMIQTVFEQLQKQPDGNWLF II II I I I I I II I I I II II I I M M I I M M I I II I II II II I I II II I I I I II I I I I I II orf81-1 IVLHQRGSHAPYGALLQPQDKVFGEADIVDKYDNTIHKTDQMIQTVFEQLQKQPDGNWLF 370 380 390 400 410 420 430 440 450 460 470 480 orf81ng-l.pep AYTSDHGQYVRQDIYNQGTVQPDSYIVPLVLYSPDKAVQQAANQAFAPCEIAFHQQLSTF I I II I II II I I I I II II I I I I II I I : I I I I II I I I I I I II II I I II I I I I I I I I I I I II I orf81-1 AYTSDHGQYVRQDIYNQGTVQPDSYLVPLVLYSPDKAVQQAANQAFAPCEIAFHQQLSTF 430 440 450 460 470 480 490 500 510 520 orf81ng-l.pep LIHTLGYDMPVSGCREGSVTGNLITGDAGSLNIRNGKAEYVYPQX I II II I II II II I I II I II I I I I II I I II II I I I : I II II I I I II orf81-1 LIHTLGYDMPVSGCREGSVTGNLITGDAGSLNIRDGKAEYVYPQX 490 500 510 520 Furthermore, ORFδlng shows significant homology to an E.coli OMP: gi 11256380 (U50906) outer membrane adherence protein-associated protein [E. coli] Length = 547 Score = 87.4 bits (213), Expect = 2e-16 Identities = 122/468 (26%), Positives = 198/468 (42%), Gaps = 70/468 (14%) Query: 25 VFGIETLPAAKMAETFA-LTFMIAALYLFARYKAS—RLLIAVFFAFSMIANNVHYAVYQ 81 VFGI L A+ A L F + + + R + RLL+A F + A ++ ++Y Sbjct: 29 VFGITNLVASSGAHMVQRLLFFVLTILWKRISSLPLRLLVAAPFVL-LTAADMSISLY- 86 Query: 82 SWMT GINYWLMLKEVTEVGSAGASMLDKLWLPALWGVAEVMLFCSLAKFRRKT 134 SW T G ++ + EV A ML ++ P L A + L + Sbjct: 87 SWCTFGTTFNDGFAISVLQSDPDEV AKMLG-MYSPYLCAFAFLSLLFLAVIIKYDV 141 Query: 135 HFSADILFAFLMLMIFVRSF DTKQEHGISPKPTYSRIKAN—YFSFGYFVG 183 + L+L++ S D K ++ SP SR +F+ YF Sbjct: 142 SLPTKKVTGILLLIVISGSLFSACQFAYKDAKNKNAFSPYILASRFATYTPFFNLNYFAL 201 Query: 184 RVLPYQ—LFDLSKIPVFKQPAPSKIGQGSIQNIVLIMGESESAAHLKLFGYGRETSPFL 241 +Q L + +P F+ + I VLI+GES ++ L+GY R T+P + Sbjct: 202 AAKEHQRLLSIANTVPYFQL SVRDTGIDTYVLIVGESVRVDNMSLYGYTRSTTPQV 257 Query: 242 TRLSQADFKPIVKQSYSAGFMTAVSLP SFFNVIPHANGLEQISGGDTNMFRLAKEQG 298 +Q + Q+ S TA+S+P + +V+ H I N+ +A + G Sbjct: 258 E—AQRKQIKLFNQAISGAPYTALSVPLSLTADSVLSH DIHNYPDNIINMANQAG 310 Query: 299 YETYFYSAQA ENQMAILNLIGKKWIDHLIQPTQLGYGNGDNMPDEKLLPLFDKINLQ 355 ++T++ S+Q+ +N A+ ++ ++ + Y G DE LLP + Q Sbjct: 311 FQTFWLSSQSAFRQNGTAVTSI AMRAMETVYVRGF DELLLPHLSQALQQ 359 Query: 356 —QGRHFIVLHQRGSHAPYGALLQPQDKVFGEADIVDK-YDNTIHKTDQMIQTVFEQLQK 412 Q + IVLH GSH P + VF D D YDN+IH TD ++ VFE L+ Sbjct: 360 NTQQKKLIVLHLNGSHEPACSAYPQSSAVFQPQDDQDACYDNSIHYTDSLLGQVFELLK- 418 Query: 413 QPDGNWLFAYTSDHG QYVRQDIYNQG—TVQPDSYIVPL-VLYSP 454 D Y +DHG ++++Y G +Y VP+ + YSP Sbjct: 419 —DRRASVMYFADHGLERDPTKKNVYFHGGREASQQAYHVPMFIWYSP 464 Based on this analysis, including the presence of a putative leader sequence (double-underlined) and several putative transmembrane domains (single-underlined) in the gonococcal protein, it is predicted that the proteins from N. meningitidis and N.gonorrhoeae, and their epitopes, could be useful antigens for vaccines or diagnostics, or for raising antibodies. Example 37 The following partial DNA sequence was identified in N. meningitidis <SEQ LD 311>: 1 ...ACCCTGCTCC TCTTCATCCC CCTCGTCCTC ACAC.GTGCG GCACACTGAC 51 CGGCATACTC GCCCaCGGCG GCGGCAAACG CTTTGCCGTC GAACAAGAAC 101 TCGTCGCCGC ATCGTCCCGC GCCGCCGTCA AAGAAATGGA TTTGTCCGCC 151 yTAAAAGGAC GCAAAGCCGC CyTTTACGTC TCCGTTATGG GCGACCAAGG 201 TTCGGGCAAC ATAAGCGGCG GACGCTACTC TATCGACGCA CTGATACGCG 251 GCGGCTACCA CAACAACCCC GAAAGTGCCA CCCAATACAG CTACCCCGCC 301 TACGACACTA CCGCCACCAC CAAATCCGAC GCGCTCTCCA GCGTAACCAC 351 TTCCACATCG CTTTTGAACG CCCCCGCCGC CGyCyTGACG AAAAACAGCG 401 GACGCAAAGG CGAACGcTCC GCCGGACTGT CCGTCAACGG CACGGGCGAC 451 TACCGCAACG AAACCCTGCT CGCCAACCCC CGCGACGTTT CCTTCCTGAC 501 CAACCTCATC CAAACCGTCT TCTACCTGCG CGGCATCGAA GTCgTACCGC 551 CCGrATACGC CGACACCGAC GTATTCGTAA CCGTCGACGT A... This corresponds to the .amino acid sequence <SEQ ID 312; ORF83>: 1 ..TLLLFIPLVL TXCGTLTGIL AHGGGKRFAV EQELVAASSR AAVKEMDLSA 51 LKGRKAAXYV SVMGDQGSGN ISGGRYSIDA LIRGGYHNNP ESATQYSYPA 101 YDTTATTKSD ALSSVTTSTS LLNAPAAXLT KNSGRKGERS AGLSVNGTGD 151 YRNETLLANP RDVSFLTNLI QTVFYLRGIE WPPXYADTD VFVTVDV.. Further work revealed the complete nucleotide sequence <SEQ ID 313>: 1 ATGAAAACCC TGCTCCTCCT CATCCCCCTC GTCCTCACAG CCTGCGGCAC 51 ACTGACCGGC ATACCCGCCC ACGGCGGCGG CAAACGCTTT GCCGTCGAAC 101 AAGAACTCGT CGCCGCATCG TCCCGCGCCG CCGTCAAAGA AATGGATTTG 151 TCCGCCCTAA AAGGACGCAA AGCCGCCCTT TACGTCTCCG TTATGGGCGA 201 CCAAGGTTCG GGCAACATAA GCGGCGGACG CTACTCTATC GACGCACTGA 251 TACGCGGCGG CTACCACAAC AACCCCGAAA GTGCCACCCA ATACAGCTAC 301 CCCGCCTACG ACACTACCGC CACCACCAAA TCCGACGCGC TCTCCAGCGT 351 AACCACTTCC ACATCGCTTT TGAACGCCCC CGCCGCCGCC CTGACGAAAA 401 ACAGCGGACG CAAAGGCGAA CGCTCCGCCG GACTGTCCGT CAACGGCACG 451 GGCGACTACC GCAACGAAAC CCTGCTCGCC AACCCCCGCG ACGTTTCCTT 501 CCTGACCAAC CTCATCCAAA CCGTCTTCTA CCTGCGCGGC ATCGAAGTCG 551 TACCGCCCGA ATACGCCGAC ACCGACGTAT TCGTAACCGT CGACGTATTC 601 GGCACCGTCC GCAGCCGTAC CGAACTGCAC CTCTACAACG CCGAAACCCT 651 TAAAGCCCAA ACCAAGCTCG AATATTTCGC CGTTGACCGC GACAGCCGGA 701 AACTGCTGAT TACCCCTAAA ACCGCCGCCT ACGAATCCCA ATACCAAGAA 751 CAATACGCCC TTTGGACCGG CCCTTACAAA GTCAGCAAAA CCGTCAAAGC 801 CTCAGACCGC CTGATGGTCG ATTTCTCCGA CATTACCCCC TACGGCGACA 851 CAACCGCCCA AAACCGTCCC GACTTCAAAC AAAACAACGG TAAAAAACCC 901 GATGTCGGCA ACGAAGTCAT CCGCCGCCGC AAAGGAGGAT AA This corresponds to the amino acid sequence <SEQ ID 314; ORF83-l>: 1 MKTLLLLIPL VLTACGTLTG IPAHGGGKRF AVEQELVAAS SRAAVKEMDL 51 SALKGRKAAL YVSVMGDQGS GNISGGRYSI DALIRGGYHN NPESATQYSY 101 PAYDTTATTK SDALSSVTTS TSLLNAPAAA LTKNSGRKGE RSAGLSVNGT 151 GDYRNETLLA NPRDVSFLTN LIQTVFYLRG IEWPPEYAD TDVFVTVDVF 201 GTVRSRTELH LYNAETLKAQ TKLEYFAVDR DSRKLLITPK TAAYESQYQE 251 QYALWTGPYK VSKTVKASDR LMVDFSDITP YGDTTAQNRP DFKQNNGKKP 301 DVGNEVIRRR KGG* Computer analysis of this amino acid sequence gave the following results: Homology with a predicted ORF from N.meninsitidis (strain A) ORF83 shows 96.4% identity over a 197aa overlap with an ORF (ORF83a) from strain A of N. meningitidis: 10 20 30 40 50 orf83.pep TLLLFIPLVLTXCGTLTGILAHGGGKRFAVEQELVAASSRAAVKEMDLSALKGRKAAX III : II II I I I I II I II II I II I I I II I II II I I II II I I I II I I I I II I I I I I orf83a MKTLLXLIPLVLTACGTLTGIPAHGGGKRFAVEQELVAASSRAAVKEMDLSALKGRKAAL 10 20 30 40 50 60 60 70 80 90 100 110 orf83. pep YVSVMGDQGSGNISGGRYSIDALIRGGYHNNPESATQYSYPAYDTTATTKSDALSSVTTS I I II II I I II II II II I II I II I I II II I II I II I II II I I I II II I II I I I II I I I I II orf83a YVSVMGDQGSGNISGGRYSIDALIRGGYHNNPESATQYSYPAYDTTATTKSDALSSVTTS 70 80 90 100 110 120 120 130 140 150 160 170 orf83. pep TSLLNAPAAXLTKNSGRKGERSAGLSVNGTGDYRNETLLANPRDVSFLTNLIQTVFYLRG I II II I II I I II I I I II I II I I II I II II I II I II I I II I I II I II I II I I II I I I II I orf83a TSLLNAPAAALTKNSGRKGERSAGLSVNGTGDYRNETLLANPRDVSFLTNLIQTVFYLRG 130 140 150 160 170 180 180 190 orf83. pep IEWPPXYADTDVFVTVDV I I II II II I II II I II I I orf83a IEWPPEYADTDVFVTVDVFGTVRSRTELHLYNAETLKAQTKLEYFAVDRDSRKLLIAPK 190 200 210 220 230 240 The complete length ORF83a nucleotide sequence <SEQ ID 315> is: 1 ATGAAAACCC TGCTCNTCCT CATCCCCCTC GTCCTCACAG CCTGCGGCAC 51 ACTGACCGGC ATACCCGCCC ACGGCGGCGG CAAACGCTTT GCCGTCGAAC 101 AAGAACTCGT CGCCGCATCG TCCCGCGCCG CCGTCAAAGA AATGGACTTG 151 TCCGCCCTGA AAGGACGCAA AGCCGCCCTT TACGTCTCCG TTATGGGCGA 201 CCAAGGTTCG GGCAACATAA GCGGCGGACG CTACTCTATC GACGCACTGA 251 TACGCGGCGG CTACCACAAC AACCCCGAAA GTGCCACCCA ATACAGCTAC 301 CCCGCCTACG ACACTACCGC CACCACCAAA TCCGACGCGC TCTCCAGCGT 351 AACCACTTCC ACATCGCTTT TGAACGCCCC CGCCGCCGCC CTGACGAAAA 401 ACAGCGGACG CAAAGGCGAA CGCTCCGCCG GACTGTCCGT CAACGGCACG 451 GGCGACTACC GCAACGAAAC CCTGCTCGCC AACCCCCGCG ACGTTTCCTT 501 CCTGACCAAC CTCATCCAAA CCGTCTTCTA CCTGCGCGGC ATCGAAGTCG 551 TACCGCCCGA ATACGCCGAC ACCGACGTAT TCGTAACCGT CGACGTATTC 601 GGCACCGTCC GCAGCCGCAC CGAACTGCAC CTCTACAACG CCGAAACCCT 651 TAAAGCCCAA ACCAAGCTCG AATATTTCGC CGTTGACCGC GACAGCCGGA 701 AACTGCTGAT TGCCCCTAAA ACCGCCGCCT ACGAATCCCA ATACCAAGAA 751 CAATACGCCC TCTGGATGGG ACCTTACAGC GTCGGCAAAA CCGTCAAAGC 801 CTCAGACCGC CTGATGGTCG ATTTCTCCGA CATCACCCCC TACGGCGACA 851 CAACCGCCCA AAACCGTCCC GACTTCAAAC AAAACAACGG TAAAAAACCC 901 GATGTCGGCA ACGAAGTCAT CCGCCGCCGC AAAGGAGGAT AA This encodes a protein having amino acid sequence <SEQ LD 316>: 1 MKTLLXLIPL VLTACGTLTG IPAHGGGKRF AVEQELVAAS SRAAVKEMDL 51 SALKGRKAAL YVSVMGDQGS GNISGGRYSI DALIRGGYHN NPESATQYSY 101 PAYDTTATTK SDALSSVTTS TSLLNAPAAA LTKNSGRKGE RSAGLSVNGT 151 GDYRNETLLA NPRDVSFLTN LIQTVFYLRG IEWPPEYAD TDVFVTVDVF 201 GTVRSRTELH LYNAETLKAQ TKLEYFAVDR DSRKLLIAPK TAAYESQYQE 251 QYALWMGPYS VGKTVKASDR LMVDFSDITP YGDTTAQNRP DFKQNNGKKP 301 DVGNEVIRRR KGG* ORF83a and ORF83-1 show 98.4% identity in 313 aa overlap: 10 20 30 40 50 60 orf83a . pep MKTLLXLIPLVLTACGTLTGIPAHGGGKRFAVEQELVAASSRAAVKEMDLSALKGRKAAL II I II I II II I II II I II II II I II II II II II II I I I I I I I I II II I II I I II I I I I I orf83-1 MKTLLLLIPLVLTACGTLTGIPAHGGGKRFAVEQELVAASSRAAVKEMDLSALKGRKAAL 10 20 30 40 50 60 70 80 90 100 110 120 orf83a . pep YVSVMGDQGSGNISGGRYSIDALIRGGYHNNPESATQYSYPAYDTTATTKSDALSSVTTS I I II II II I II II II II I II II I I II I I I I II II II I I II II II I I II I II I I II I I I I I orf83-1 YVSVMGDQGSGNISGGRYSIDALIRGGYHNNPESATQYSYPAYDTTATTKSDALSSVTTS 70 80 90 100 110 120 130 140 150 160 170 180 orf83a . pep TSLLNAPAAALTKNSGRKGERSAGLSVNGTGDYRNETLLANPRDVSFLTNLIQTVFYLRG I I II II I I I II I I II II II I I I I I II II II II II II II I II II I II II I I I I I I II II II orf83-1 TSLLNAPAAALTKNSGRKGERSAGLSVNGTGDYRNETLLANPRDVSFLTNLIQTVFYLRG 130 140 150 160 170 180 190 200 210 220 230 240 orf83a . pep IEWPPEYADTDVFVTVDVFGTVRSRTELHLYNAETLKAQTKLEYFAVDRDSRKLLIAPK II I II I II II II II II II II II I II I II II II II II II I II I I I II I II I I II I I I I: I I orf83-1 IEWPPEYADTDVFVTVDVFGTVRSRTELHLYNAETLKAQTKLEYFAVDRDSRKLLITPK 190 200 210 220 230 240 250 260 270 280 290 300 orf83a . pep TAAYESQYQEQYALWMGPYSVGKTVKASDRLMVDFSDITPYGDTTAQNRPDFKQNNGKKP I I II I II II I II II I II I : I : I II I II II II I I II I I I I I I I II I I II II I I II I I I I I orf83-1 TAAYESQYQEQYALWTGPYKVSKTVKASDRLMVDFSDITPYGDTTAQNRPDFKQNNGKKP 250 260 270 280 290 300 310 orf83a. pep DVGNEVIRRRKGGX I II I II I I I II I I I orf83-l DVGNEVIRRRKGGX 310 Homology with a predicted ORF from N.sonorrhoeae ORF83 shows 94.9% identity over a 197aa overlap with a predicted ORF (ORF83.ng) from N. gonorrhoeae: orf83.pep TLLLFIPLVLTXCGTLTGILAHGGGKRFAVEQELVAASSRAAVKEMDLSALKGRKAAX 58 II I I : I I I I I I I II II II II II I II I I I I II I II II II I I I I II II II I II I I I I orf83ng MKTLLLLIPLVLTACGTLTGIPAHGGGKRFAVEQELVAASSRAAVKEMDLSALKGRKAAL 60 orf83.pep YVSVMGDQGSGNISGGRYSIDALIRGGYHNNPESATQYSYPAYDTTATTKSDALSSVTTS 118 I I I II II II I I II II I II II II I II II I I I II : I II: I I I I I II I II II I I I II I: II II orf83ng YVSVMGDQGSGNISGGRYSIDALIRGGYHNNPDSATRYSYPAYDTTATTKSDALSGVTTS 120 orf83. pep TSLLNAPAAXLTKNSGRKGERSAGLSVNGTGDYRNETLLANPRDVSFLTNLIQTVFYLRG 178 I I I II II II II I I : I II I I I I I II I II I I II I I I II I I I I I II I II I II I II I I I I I I I orf83ng TSLLNAPAAALTKNNGRKGERSAGLSVNGTGDYRNETLLANPRDVSFLTNLIQTVFYLRG 180 orf83. pep IEWPPXYADTDVFVTVDV 197 I I II I I II I II I II I II I orf83ng IEWPPEYADTDVFVTVDVFGTVRSRTELHLYNAETLKAQTKLEYFAVDRDSRKLLIAPK 240 The complete length ORF83ng nucleotide sequence <SEQ ID 317> is: 1 ATGAAAACCC TGCTCCTCCT CATCCCCCTC GTACTCACCG CCTGCGGCAC 51 ACTGACCGGC ATACCCGCCC ACGGCGGCGG CAAACGCTTT GCCGTCGAAC 101 AGGAACTCGT CGCCGCATCG TCCCGCGCCG CCGTCAAAGA AATGGACTTG 151 TCCGCCCTGA AAGGACGCAA AGCCGCCCTT TACGTCTCCG TTATGGGCGA 201 CCAAGGTTCG GGCAACATAA GCGGCGGACG CTACTCCATC GACGCACTGA 251 TACGCGGCGG CTACCACAAC AACCCCGACA GCGCCACCCG ATACAGCTAC 301 CCCGCCTATG ACACTACCGC CACCACCAAA TCCGACGCGC TCTCCGGCGT 351 AACCACTTCC ACATCGCTTT TGAACGCCCC CGCCGCCGCC CTGACGAAAA 401 ACAACGGACG CAAAGGCGAA CGCTCCGCCG GACTGTCCGT CAACGGCACG 451 GGCGACTACC GCAACGAAAC CCTGCTCGCC AACCCCCGCG ACGTTTCCTT 501 CCTGACCAAC CTCATCCAAA CCGTCTTCTA CCTGCGCGGC ATCGAAGTCG 551 TACCGCCCGA ATACGCCGAC ACCGACGTAT TCGTAACCGT CGACGTATTC 601 GGCACCGTCC GCAGCCGTAC CGAACTGCAC CTCTACAACG CCGAAACCCT 651 TAAAGCCCAA ACCAAGCTCG AATATTTCGC CGTCGACCGC GACAGCCGGA 701 AACTGCTGAT TGCCCCTAAA ACCGCCGCCT ACGAATCCCA ATACCAAGAA 751 CAATACGCCC TCTGGATGGG ACCTTACAGC GTCGGCAAAA CCGTCAAAGC 801 CTCAGACCGC CTGATGGTCG ATTTCTCCGA CATCACCCCC TACGGCGACA 851 CAACCGCCCA AAACCGTCCC GACTTCAAAC AAAACAACGG TAAAAACCCC 901 GATGTCGGCA ACGAAGTCAT CCGCCGCCGC AAAGGAGGAT AA This encodes a protein having amino acid sequence <SEQ ID 318>: 1 MKTLLLLIPL VLTACGTLTG IPAHGGGKRF AVEQELVAAS SRAAVKEMDL 51 SALKGRKAAL YVSVMGDQGS GNISGGRYSI DALIRGGYHN NPDSATRYSY 101 PAYDTTATTK SDALSGVTTS TSLLNAPAAA LTKNNGRKGE RSAGLSVNGT 151 GDYRNETLLA NPRDVSFLTN LIQTVFYLRG IEWPPEYAD TDVFVTVDVF 201 GTVRSRTELH LYNAETLKAQ TKLEYFAVDR DSRKLLIAPK TAAYESQYQE 251 OYALWMGPYS VGKTVKASDR LMVDFSDITP YGDTTAQNRP DFKQNNGKNP 301 DVGNEVIRRR KGG* ORF83ng and ORF83-1 show 97.1% identity in 313 aa overlap 10 20 30 40 50 60 orf83-1. pep MKTLLLLIPLVLTACGTLTGIPAHGGGKRFAVEQELVAASSRAAVKEMDLSALKGRKAAL I II I I II I II I II I I I II II I II I II I I II II I I I I I I I I II II I I I II II I I II I I II I orf83ng MKTLLLLIPLVLTACGTLTGIPAHGGGKRFAVEQELVAASSRAAVKEMDLSALKGRKAAL 10 20 30 40 50 60 70 80 90 100 110 120 orf83-1. pep YVSVMGDQGSGNISGGRYSIDALIRGGYHNNPESATQYSYPAYDTTATTKSDALSSVTTS II I I I I II I I II I I I I I I II I II I II I I II IM I I M I II II II I I I II I II M II orf83ng YVSVMGDQGSGNISGGRYSIDALIRGGYHNNPDSATRYSYPAYDTTATTKSDALSGVTTS 70 80 90 100 110 120 130 140 150 160 170 180 orf83-1. pep TSLLNAPAAALTKNSGRKGERSAGLSVNGTGDYRNETLLANPRDVSFLTNLIQTVFYLRG I I II I I I I I II I IM I I II I II I I I I I II I II I I II I II I I I II II I I I II I II II II II orf83ng TSLLNAPAAALTKNNGRKGERSAGLSVNGTGDYRNETLLANPRDVSFLTNLIQTVFYLRG 130 140 150 160 170 180 190 200 210 220 230 240 orf83-l. pep IEWPPEYADTDVFVTVDVFGTVRSRTELHLYNAETLKAQTKLEYFAVDRDSRKLLITPK II I I II II II I II I I I II I II I I I II I II II II I II I I I I I II I I II II I I I I II I I: I I orf83ng IEWPPEYADTDVFVTVDVFGTVRSRTELHLYNAETLKAQTKLEYFAVDRDSRKLLIAPK 190 200 210 220 230 240 250 260 270 280 290 300 orf83-1. pep TAAYESQYQEQYALWTGPYKVSKTVKASDRLMVDFSDITPYGDTTAQNRPDFKQNNGKKP orf83ng TAAYESQYQEQYALWMGPYSVGKTVKASDRLMVDFSDITPYGDTTAQNRPDFKQNNGKNP 250 260 270 280 290 300 310 orf83-1. pep DVGNEVIRRRKGGX II I I I II I I II I II orf83ng DVGNEVIRRRKGGX 310 Based on this analysis, including the presence of a putative ATP/GTP-binding site motif A (P-loop) in the gonococcal protein (double-underlined) and a putative prokaryotic membrane lipoprotein lipid attachment site (single-underlined), it is predicted that the proteins from N. meningitidis .and N.gonorrhoeae, .and their epitopes, could be useful antigens for vaccines or diagnostics, or for raising antibodies. Example 38 The following DNA sequence, believed to be complete, was identified in N. meningitidis <SEQ ID 319>: 1 ATGGCAGAGA TCTGTTTGAT AACCGGCACG CCCGGTTCAG GGAAAACATT 51 AAAAATGGTT TCCATGATGG CGAATGATGA AATGTTTAAG CCTGATGAAA 101 AAGCCATACG CCGTAAAGTA TTTACGAACA TAAAAGGCTT GAAAATACCG 151 CACACCTACA TAGAAACGGA CGCAAAAAAG CTGCCGAAAT CGACAGATGA 201 GCAGCTTTCG GCGCATGATA TGTACGAATG GATAAAGAAG CCCGAAAATA 251 TCGGGTCTAT TGTCATTGTA GATGAAGCTC AAGACGTATG GCCGGCACGC 301 TCGGCAGGTT CAAAAATCCC TGAAAATGTC CAATGGCTGA ATACGCACAG 351 ACATCAGGGC ATTGATATAT TTGTTTTGAC TCAAGGTCCT AAGCTTCTAG 401 ATCAAAATCT TAGAACGCTT GTACGGAAAC ATTACCACAT CGCTTCAAAC 451 AAGATGGGTA TGCGTACGCT TTTAGAATGG AAAATATGCG CGGACGATCC 501 CGTAAAAATG GCATCAAGCG CATTCTCCAG TATCTATACA CTGGATAAAA 551 AAGTTTATGA CTTGTAysrr TmmGCGGAAG TTCATACCGT AAATAAGGTC 601 AAGCGGTCAA AGTGGTTTTA CACTCTGCCa GTAATAGTAT TGCTGATTCC 651 CGTGTTTGTC GGCCTGTCCT ATAAAATGTT GagCaGTTAC GGAAAAAAAC 701 aGGAAGAACC CGCAGCACAA GAATCGGCGG CAACAGAACA GCAGGCAGTA 751 CTTCCGGATA AAACAGAAGG CGAGCCGGTA AATAACGGCA ACCTTACCGC 801 AGATATGTTT GTTCCGACAT TGTCCGAaAA ACCCGrAAGC AAGCcgaTTT 851 ATAACGGTGT AAGGCAGGTA AGAACCTTTG AATATATAGC AGGCTGTATA 901 GAAGGCGGAA GAACCGGATG CGCCTGCTAT TCGCaTCAAG GGACGGCATt 951 gaAAGAAGTG ACGGaGTTGA TGTGc≤aAgG aCTATGTaAA AAacGGCTTG 1001 CCGTTTAACC CaTACAAAGA AGAAAGCCAA GGGCAGGAAG TTCAGCAAAG 1051 CGCGCAgCAA CATTCGGACA GGGCGcCAAG TTGCCACATT GGGCGGAAAA 1101 CCGTAGCAGA ACCTAATGTA CGATAATTGG GAAGAACGCG GGAAACCGTT 1151 TGAAGGAATC GGaCGGGGGC GTGGTCGGAT CGGCAAACTG A This corresponds to the amino acid sequence <SEQ ID 320; ORF84>: 1 MAEICLITGT PGSGKTLKMV SMMANDEMFK PDEKAIRRKV FTNIKGLKIP 51 HTYIETDAKK LPKSTDEQLS AHDMYEWIKK PENIGSIVIV DEAQDVWPAR 101 SAGSKIPENV QWLNTHRHQG IDIFVLTQGP KLLDQNLRTL VRKHYHIASN 151 KMGMRTLLEW KICADDPVKM ASSAFSSIYT LDKKVYDLYX XAEVHTVNKV 201 KRSKWFYTLP VIVLLIPVFV GLSYKMLSSY GKKQEEPAAQ ESAATEQQAV 251 LPDKTEGEPV NNGNLTADMF VPTLSEKPXS KPIYNGVRQV RTFEYIAGCI 301 EGGRTGCACY SHQGTALKEV TELMCKDYVK NGLPFNPYKE ESQGQEVQQS 351 AQQHSDRAQV ATLGGKPXQN LMYDNWEERG KPFEGIGGGV VGSAN* Further work revealed the complete nucleotide sequence <SEQ LD 321>: 1 ATGGCAGAGA TCTGTTTGAT AACCGGCACG CCCGGTTCAG GGAAAACATT 51 AAAAATGGTT TCCATGATGG CGAATGATGA AATGTTTAAG CCTGATGAAA 101 ACGGCATACG CCGTAAAGTA TTTACGAACA TAAAAGGCTT GAAAATACCG 151 CACACCTACA TAGAAACGGA CGCAAAAAAG CTGCCGAAAT CGACAGATGA 201 GCAGCTTTCG GCGCATGATA TGTACGAATG GATAAAGAAG CCCGAAAATA 251 TCGGGTCTAT TGTCATTGTA GATGAAGCTC AAGACGTATG GCCGGCACGC 301 TCGGCAGGTT CAAAAATCCC TGAAAATGTC CAATGGCTGA ATACGCACAG 351 ACATCAGGGC ATTGATATAT TTGTTTTGAC TCAAGGTCCT AAGCTTCTAG 401 ATCAAAATCT TAGAACGCTT GTACGGAAAC ATTACCACAT CGCTTCAAAC 451 AAGATGGGTA TGCGTACGCT TTTAGAATGG AAAATATGCG CGGACGATCC 501 CGTAAAAATG GCATCAAGCG CATTCTCCAG TATCTATACA CTGGATAAAA 551 AAGTTTATGA CTTGTACGAA TCAGCGGAAG TTCATACCGT AAATAAGGTC 601 AAGCGGTCAA AGTGGTTTTA CACTCTGCCA GTAATAGTAT TGCTGATTCC 651 CGTGTTTGTC GGCCTGTCCT ATAAAATGTT GAGCAGTTAC GGAAAAAAAC 701 AGGAAGAACC CGCAGCACAA GAATCGGCGG CAACAGAACA GCAGGCAGTA 751 CTTCCGGATA AAACAGAAGG CGAGCCGGTA AATAACGGCA ACCTTACCGC 801 AGATATGTTT GTTCCGACAT TGTCCGAAAA ACCCGAAAGC AAGCCGATTT 851 ATAACGGTGT AAGGCAGGTA AGAACCTTTG AATATATAGC AGGCTGTATA 901 GAAGGCGGAA GAACCGGATG CGCCTGCTAT TCGCATCAAG GGACGGCATT 951 GAAAGAAGTG ACGGAGTTGA TGTGCAAGGA CTATGTAAAA AACGGCTTGC 1001 CGTTTAACCC ATACAAAGAA GAAAGCCAAG GGCAGGAAGT TCAGCAAAGC 1051 GCGCAGCAAC ATTCGGACAG GGCGCAAGTT GCCACATTGG GCGGAAAACC 1101 GTAGCAGAAC CTAATGTACG ATAATTGGGA AGAACGCGGG AAACCGTTTG 1151 AAGGAATCGG CGGGGGCGTG GTCGGATCGG CAAACTGA This corresponds to the amino acid sequence <SEQ LD 322; ORF84-1 >: 1 MAEICLITGT PGSGKTLKMV SMMANDEMFK PDENGIRRKV FTNIKGLKIP 51 HTYIETDAKK LPKSTDEQLS AHDMYEWIKK PENIGSIVIV DEAQDVWPAR 101 SAGSKIPENV QWLNTHRHQG IDIFVLTQGP KLLDQNLRTL VRKHYHIASN 151 KMGMRTLLEW KICADDPVKM ASSAFSSIYT LDKKVYDLYE SAEVHTVNKV 201 KRSKWFYTLP VIVLLIPVFV GLSYKMLSSY GKKQEEPAAQ ESAATEQQAV 251 LPDKTEGEPV NNGNLTADMF VPTLSEKPES KPIYNGVRQV RTFEYIAGCI 301 EGGRTGCACY SHQGTALKEV TELMCKDYVK NGLPFNPYKE ESQGQEVQQS 351 AQQHSDRAQV ATLGGKP*QN LMYDNWEERG KPFEGIGGGV VGSAN* Computer analysis of this amino acid sequence gave the following results: Homology with a predicted ORF from N.meninsitidis (strain A) ORF84 shows 93.9% identity over a 395aa overlap with an ORF (ORF84a) from strain A of N. meningitidis: 10 20 30 40 50 60 orf84.pep MAEICLITGTPGSGKTLKMVSMMANDEMFKPDEKAIRRKVFTNIKGLKIPHTYIETDAKK I II I I I II I II I I I II I II II I II II I I I II IM: II II II II I I II I II II II II II II orf84a MAEICLITGTPGSGKTLKMVSMMANDEMFKPDENGIRRKVFTNIKGLKIPHTYIETDAKK 10 20 30 40 50 60 70 80 90 100 110 120 orf84. pep LPKSTDEQLSAHDMYEWIKKPENIGSIVIVDEAQDVWPARSAGSKIPENVQWLNTHRHQG II I I II I II II I I I II I II II I II I I I I I II II II I II I II I I II II II I I I I I I I I I I I orf84a LPKSTDEQLSAHDMYEWIKKPENIGSIVIVDEAQDVWPARSAGSKIPENVQWLNTHRHQG 70 80 90 100 110 120 130 140 150 160 170 180 orf84. pep IDIFVLTQGPKLLDQNLRTLVRKHYHIASNKMGMRTLLEWKICADDPVKMASSAFSSIYT II I II I I I I I I I II II II II I II I I I I I II II I II II II I I I II II II I I I I I I I I I I I orf84a IDIFVLTQGSKLLDQNLRTLVRKHYHIASNKMGMRTLLEWKICADDPVKMASSAFSSIYT 130 140 150 160 170 180 190 200 210 220 230 240 orf84.pep LDKKVYDLYXXAEVHTVNKVKRSKWFYTLPVIVLLIPVFVGLSYKMLSSYGKKQEEPAAQ II I II I I I I I I I II I I I I I I I II I I II I II : I I I II I I II I II I II I I I II I I I I I I I orf84a LDKKVYDLYESAEVHTVNKVKRSKWFYTLPVIILLIPVFVGLSYKMLSSYGKKQEEPAAQ 190 200 210 220 230 240 250 260 270 280 290 300 orf84. pep ESAATEQQAVLPDKTEGEPVNNGNLTADMFVPTLSEKPXSKPIYNGVRQVRTFEYIAGCI I II I II : II I : II I I II II I II I II I I I II II II II I II II I I II II I I I I I I I I I I : orf84a ESAATEHQAVFQDKTEGEPVNNGNLTADMFVPTLSEKPESKPIYNGVRQVRTFEYIAGCV 250 260 270 280 290 300 310 320 330 340 350 360 orf84. pep EGGRTGCACYSHQGTALKEVTELMCKDYVKNGLPFNPYKEESQGQEVQQSAQQHSDRAQV I I I I I I I : I II I I I I I II I : I '■ I I I I I :: I I I II II II I I II I :: I II I MM II orf84a EGGRTGCTCYSHQGTALKEITKEMCKDYARNGLPFNPYKEESQGRDVQQSEQHHSDRPQV 310 320 330 340 350 360 370 380 390 orf84.pep ATLGGKPXQNLMYDNWEERGKPFEGIGGGWGSANX I I II I I I II I I I I I I: I II I I II II II I II I I I II orf84a ATLGGKPWQNLMYDNWQERGKPFEGIGGGWGSANX 370 380 390 The complete length ORF84a nucleotide sequence <SEQ ID 323> is: 1 ATGGCAGAGA TCTGTTTGAT AACCGGCACG CCCGGTTCAG GGAAAACATT 51 AAAAATGGTT TCCATGATGG CAAACGATGA AATGTTTAAG CCGGATGAAA 101 ACGGCATACG CCGTAAAGTA TTTACGAACA TCAAAGGCTT GAAGATACCG 151 CACACCTACA TAGAAACGGA CGCGAAAAAG CTGCCGAAAT CGACAGATGA 201 GCAGCTTTCG GCGCATGATA TGTACGAATG GATAAAGAAG CCCGAAAATA 251 TCGGGTCTAT TGTCATTGTA GATGAAGCTC AAGACGTATG GCCGGCACGC 301 TCGGCAGGTT CAAAAATCCC TGAAAATGTC CAATGGCTGA ATACGCACAG 351 ACATCAGGGC ATTGATATAT TTGTTTTGAC TCAAGGCTCT AAGCTTCTAG 401 ATCAAAATCT TAGAACGCTT GTACGGAAAC ATTACCACAT CGCTTCAAAC 451 AAGATGGGTA TGCGTACGCT TTTAGAATGG AAAATATGCG CGGACGATCC 501 CGTAAAAATG GCATCAAGCG CATTCTCCAG TATCTATACA CTGGATAAAA 551 AAGTTTATGA CTTGTACGAA TCAGCGGAAG TTCATACCGT AAATAAGGTC 601 AAGCGGTCAA AATGGTTTTA TACTCTGCCA GTAATAATAT TGCTGATTCC 651 CGTTTTTGTC GGCCTGTCCT ATAAAATGTT AAGTAGTTAT GGAAAAAAAC 701 AGGAAGAACC CGCAGCACAA GAATCGGCGG CAACAGAACA TCAGGCAGTA 751 TTTCAGGATA AAACAGAAGG CGAGCCGGTA AACAACGGTA ACCTTACCGC 801 AGATATGTTT GTTCCGACAT TGTCCGAAAA ACCCGAAAGC AAGCCGATTT 851 ATAACGGTGT AAGGCAGGTA AGAACCTTTG AATATATAGC AGGCTGTGTA 901 GAAGGCGGAA GAACCGGATG CACATGCTAT TCGCATCAAG GGACGGCATT 951 GAAAGAAATT ACAAAGGAAA TGTGCAAGGA TTACGCAAGA AACGGATTGC 1001 CGTTTAACCC ATATAAAGAA GAAAGCCAAG GGCGGGATGT CCAGCAAAGT 1051 GAGCAGCACC ATTCGGACAG ACCGCAAGTT GCCACGTTGG GCGGAAAGCC 1101 GTGGCAAAAT CTTATGTATG ATAATTGGCA GGAGCGCGGA AAACCGTTTG 1151 AAGGAATCGG CGGGGGCGTG GTCGGATCGG CAAACTGA This encodes a protein having amino acid sequence <SEQ ID 324>: 1 MAEICLITGT PGSGKTLKMV SMMANDEMFK PDENGIRRKV FTNIKGLKIP 51 HTYIETDAKK LPKΞTDEQLS AHDMYEWIKK PENIGSIVIV DEAQDVWPAR 101 SAGSKIPENV QWLNTHRHQG IDIFVLTQGS KLLDQNLRTL VRKHYHIASN 151 KMGMRTLLEW KICADDPVKM ASSAFSSIYT LDKKVYDLYE SAEVHTVNKV 201 KRSKWFYTLP VIILLIPVFV GLSYKMLSSY GKKQEEPAAQ ESAATEHQAV 251 FQDKTEGEPV NNGNLTADMF VPTLSEKPES KPIYNGVRQV RTFEYIAGCV 301 EGGRTGCTCY SHQGTALKEI TKEMCKDYAR NGLPFNPYKE ESQGRDVQQS 351 EQHHSDRPQV ATLGGKPWQN LMYDNWQERG KPFEGIGGGV VGSAN* ORF84a and ORF84-1 show 95.2% identity in 395 aa overlap: 10 20 30 40 50 60 orf84a . pep MAEICLITGTPGSGKTLKMVSMMANDEMFKPDENGIRRKVFTNIKGLKIPHTYIETDAKK I I I II I II I II I I I II II I II II I I II II I I II I II I II II I I II I II II I II I I I I II I orf84-1 MAEICLITGTPGSGKTLKMVSMMANDEMFKPDENGIRRKVFTNIKGLKIPHTYIETDAKK 10 20 30 40 50 60 70 80 90 100 110 120 orf84a. pep LPKSTDEQLSAHDMYEWIKKPENIGSIVIVDEAQDVWPARSAGSKIPENVQWLNTHRHQG I II I I I I I I I I I I I I II I I II II II I II I I II I I II II II II I II II I I I 1 II I II II I I orf84-1 LPKSTDEQLSAHDMYEWIKKPENIGSIVIVDEAQDVWPARSAGSKIPENVQWLNTHRHQG 70 80 90 100 110 120 130 140 150 160 170 180 orf84a.pep IDIFVLTQGSKLLDQNLRTLVRKHYHIASNKMGMRTLLEWKICADDPVKMASSAFSSIYT I I II I II I I I II I II I II I I I II I II II II I I II I II I I I II I I I II I I I II I II I I I I orf84-1 IDIFVLTQGPKLLDQNLRTLVRKHYHIASNKMGMRTLLEWKICADDPVKMASSAFSSIYT 130 140 150 160 170 180 190 200 210 220 230 240 orf84a. pep LDKKVYDLYESAEVHTVNKVKRSKWFYTLPVIILLIPVFVGLSYKMLSSYGKKQEEPAAQ M I I II I II I II I II II I I II II I II I I I I II : I I II II II I I I I II I I II I II I I I I II orf84-1 LDKKVYDLYESAEVHTVNKVKRSKWFYTLPVIVLLIPVFVGLSYKMLSSYGKKQEEPAAQ 190 200 210 220 230 240 250 260 270 280 290 300 orf84a. pep ESAATEHQAVFQDKTEGEPVNNGNLTADMFVPTLSEKPESKPIYNGVRQVRTFEYIAGCV I II I II: II I: I I I I II I II II I II I I I II I II I II I I I I II II II II I I II I II II I : orf84-1 ESAATEQQAVLPDKTEGEPVNNGNLTADMFVPTLSEKPESKPIYNGVRQVRTFEYIAGCI 250 260 270 280 290 300 310 320 330 340 350 360 orf84a . pep EGGRTGCTCYSHQGTALKEITKEMCKDYARNGLPFNPYKEESQGRDVQQSEQHHSDRPQV 1 I I I I I I : I I II I II I I I MM I I II I :: I I I II I II I I I II I :: I I I I MM II orf84-1 EGGRTGCACYSHQGTALKEVTELMCKDYVKNGLPFNPYKEESQGQEVQQSAQQHSDRAQV 310 320 330 340 350 360 370 380 390 orf84a.pep ATLGGKPWQNLMYDNWQERGKPFEGIGGGWGSANX I II II II I II I II I I: I II II I I I II I I II I II I I orf84-1 ATLGGKPXQNLMYDNWEERGKPFEGIGGGWGSANX 370 380 390 Homology with a predicted ORF from N.sonorrhoeae ORF84 shows 94.2% identity over a 395aa overlap with a predicted ORF (ORF84.ng) from N gonorrhoeae: orf 84 . pep MAEICLITGTPGSGKTLKMVSMMAΝDEMFKPDEKAIRRKVFTΝIKGLKIPHTYIETDAKK 60 I I I II I I I I I II II II I I I II I I I II II II I I I::: II I I I I II I II I I I II : I II II I I orf84ng MAEICLITGTPGSGKTLKMVSMMAΝDEMFKPDEΝGVRRKVFTΝIKGLKIPHTHIETDAKK 60 orf84.pep LPKSTDEQLSAHDMYEWIKKPEΝIGSIVIVDEAQDVWPARSAGSKIPEΝVQWLΝTHRHQG 120 I II I II II I II II II II II I I II : I : II II I I I I II II II I I II I I I I I I II I I I II II I orf84ng LPKSTDEQLSAHDMYEWIKKPEΝVGAIVIVDEAQDVWPARSAGSKIPEΝVQWLΝTHRHQG 120 orf84.pep IDIFVLTQGPKLLDQΝLRTLVRKHYHIASΝKMGMRTLLEWKICADDPVKMASSAFSSIYT 180 II I I I I I I I I I II II II I II I :: I I I II : I I I I : I I II I II : II I I I I I II I II I II II I orf84ng IDIFVLTQGPKLLDQΝLRTLVKRHYHIAAΝKMGLRTLLEWKVCADDPVKMASSAFSSIYT 180 orf84.pep LDKKVYDLYXXAEVHTVΝKVKRSKWFYTLPVIVLLIPVFVGLSYKMLSSYGKKQEEPAAQ 240 I I I II I I I I I I : I I I I I I I II I I I I : I I I I : I II I : I II I II I I I : II I I II I I I I II orf84ng LDKKVYDLYESAEIHTVΝKVKRSKWFYALPVIILLIPLFVGLSYKMLGSYGKKQEEPAAQ 240 orf84.pep ESAATEQQAVLPDKTEGEPVΝΝGΝLTADMFVPTLSEKPXSKPIYΝGVRQVRTFEYIAGCI 300 I I II I I I I I I II II I I II I II I I I II I I I I I I I III II I II I II I I II I II I I I II I orf84ng ESAATEQQAVLPDKTEGESVΝΝGΝLTADMFVPTLPEKPESKPIYΝGVRQVRTFEYIAGCI 300 orf84.pep EGGRTGCACYSHQGTALKEVTELMCKDYVKΝGLPFΝPYKEESQGQEVQQSAQQHSDRAQV 360 I I II I I I : I I I I I I I I II II II I I I I II II I I I I II II II I I II I I I I II I I II I I I I II orf84ng EGGRTGCTCYSHQGTALKEVTELMCKDYVKΝGLPFΝPYKEESQGQEVQQSAQQHSDRAQV 360 orf84.pep ATLGGKPXQΝLMYDΝWEERGKPFEGIGGGWGSAΝ 395 I II I I I I I I II II I I II II II I I I I II II I I I I I orf84ng ATLGGKPQQΝLMYDΝWEERGKPFEGIGGGWGSAΝ 395 The complete length ORF84ng nucleotide sequence <SEQ ID 325> is: 1 ATGGCAGAAA TCTGTTTGAT AACCGGCACG CCCGGTTCAG GGAAAACATT 51 AAAAATGGTT TCCATGATGG CAAACGATGA AATGTTTAAG CCAGATGAAA 101 ACGGCGTACG CCGTAAAGTA TTTACGAACA TCAAAGGTTT GAAGATACCG 151 CACACCCACA TAGAAACAGA CGCAAAGAAG CTGCCGAAAT CAACCGATGA 201 ACAGCTTTCG GCGCATGATA TGTATGAATG GATCAAGAAG CCTGAAAacg 251 tcggcgCAAT CGTTATTGTC GATGAGGCGC AAGACGTATG GCCCGCACGC 301 TccgCAGGTT CGAAAATCCC CGAAAACGTC CAATGGCTGA ACACACACAG 351 GCATCAGGGC ATAGATATAT TTGTATTGAC ACAAGGTCCT AAACTCTTAG 401 ATCAGAACTT GCGAACATTG GTTAAAAGAC ATTACCACAT TGCGGCCAAC 451 AAAATGGGTT TGCGTACCCT GCTTGAATGG AAAGTATGCG CGGATGACCC 501 CGTAAAAATG GCATCAAGTG CATTTTCCAG TATCTACACA CTGGATAAAA 551 AAGTTTATGA CTTGTACGAA TCCGCAGAAA TTCACACGGT AAACAAAGTC 601 AAGCGTTCAA AATGGTTTTA TGCATTGCCC GTCATCATAT TATTGATTCC 651 GCTATTTGTC GGTTTGTCTT ACAAAATGTT GGGCAGTTAC GGAAAAAAAC 701 AGGAAGAACC CGCAGCACAA GAATCGGCGG CAACAGAACA GCAGGCAGTA 751 CTTCCGGATA AAACAGAAGG AGAATCGGTG AATAACGGAA ACCTTACGGC 801 AGATATGTTT GTTCCGACAT TGCCCGAAAA ACCCGAAAGC AAGCCGATTT 851 ATAACGGTGT AAGGCAGGTA AGGACCTTTG AATATATAGC AGGCTGTATA 901 GAAGGCGGAA GAACCGGATG CACCTGCTAT TCGCATCAAG GGACGGCATT 951 GAAAGAAGTG ACGGAGTTGA TGTGCAAGGA CTATGTAAAA AACGGCTTGC 1001 CGTTTAACCC ATACAAAGAA GAAAGCCAAG GGCAGGAAGT TCAGCAAAGC 1051 GCGCAGCAAC ATTCGGACAG GGCGCAAGTT GCCACCTTGG GCGGAAAACC 1101 GCAGCAGAAC CTAATGTACG ACAATTGGGA AGAACGCGGG AAACCGTTTG 1151 AAGGAATCGG CGGGGGCGTG GTCGGATCGG CAAACTGA This encodes a protein having amino acid sequence <SEQ ID 326>: 1 MAEICLITGT PGSGKTLKMV SMMAΝDEMFK PDEΝGVRRKV FTΝIKGLKIP 51 HTHIETDAKK LPKSTDEQLS AHDMYEWIKK PEΝVGAIVIV DEAQDVWPAR 101 SAGSKIPEΝV QWLΝTHRHQG IDIFVLTQGP KLLDQΝLRTL VKRHYHIAAΝ 151 KMGLRTLLEW KVCADDPVKM ASSAFSSIYT LDKKVYDLYE SAEIHTVΝKV 201 KRSKWFYALP VIILLIPLFV GLSYKMLGSY GKKQEEPAAQ ESAATEQQAV 251 LPDKTEGESV ΝΝGΝLTADMF VPTLPEKPES KPIYΝGVRQV RTFEYIAGCI 301 EGGRTGCTCY SHQGTALKEV TELMCKDYVK ΝGLPFΝPYKE ESQGQEVQQS 351 AQQHSDRAQV ATLGGKPQQΝ LMYDΝWEERG KPFEGIGGGV VGSAΝ* ORF84ng and ORF84-1 show 95.4% identity in 395 aa overlap: 10 20 30 40 50 60 orf84-1. pep MAEICLITGTPGSGKTLKMVSMMANDEMFKPDENGIRRKVFTNIKGLKIPHTYIETDAKK II II I I II I II I II I I II II I I II II I II I I I II I : I II I I I II I II I I I I I : I I II I I I orf84ng MAEICLITGTPGSGKTLKMVSMMANDEMFKPDENGVRRKVFTNIKGLKIPHTHIETDAKK 10 20 30 40 50 60 70 80 90 100 110 120 orf84-1. pep LPKSTDEQLSAHDMYEWIKKPENIGSIVIVDEAQDVWPARSAGSKIPENVQWLNTHRHQG I I II II II II I I II II II II I II : I : I II I II II I II II I I I II I I I I I I I II II I II I I orf84ng LPKSTDEQLSAHDMYEWIKKPENVGAIVIVDEAQDVWPARSAGSKIPENVQWLNTHRHQG 70 80 90 100 110 120 130 140 150 160 170 180 orf84-1.pep IDIFVLTQGPKLLDQNLRTLVRKHYHIASNKMGMRTLLEWKICADDPVKMASSAFSSIYT I II II I II II I I II I I II II I :: I I II I : I II I : I II II II : I I II I I II II I I II I I II orf84ng IDIFVLTQGPKLLDQNLRTLVKRHYHIAANKMGLRTLLEWKVCADDPVKMASSAFSSIYT 130 140 150 160 170 180 190 200 210 220 230 240 orf84-1. pep LDKKVYDLYESAEVHTVNKVKRSKWFYTLPVIVLLIPVFVGLSYKMLSSYGKKQEEPAAQ II I I I II I I I I I I: II II II I I II I MM I II: II II: I II II II I I : I I II II II II I I orf84ng LDKKVYDLYESAEIHTVNKVKRSKWFYALPVIILLIPLFVGLSYKMLGSYGKKQEEPAAQ 190 200 210 220 230 240 250 260 270 280 290 300 orf84-1. pep ESAATEQQAVLPDKTEGEPVNNGNLTADMFVPTLSEKPESKPIYNGVRQVRTFEYIAGCI I I I I I I I I I I I I I I II II I I I I I I II I II II I I I I I I I I II II I I I I I I I II I I II I I orf84ng ESAATEQQAVLPDKTEGESVNNGNLTADMFVPTLPEKPESKPIYNGVRQVRTFEYIAGCI 250 260 270 280 290 300 310 320 330 340 350 360 orf84-1. pep EGGRTGCACYSHQGTALKEVTELMCKDYVKNGLPFNPYKEESQGQEVQQSAQQHSDRAQV I I II I I I: I II II II I I I II I I II I II I II I I I II I I I II I II II II I I I I I I II I I II I orf84ng EGGRTGCTCYSHQGTALKEVTELMCKDYVKNGLPFNPYKEESQGQEVQQSAQQHSDRAQV 310 320 330 340 350 360 370 380 390 orf84-1.pep ATLGGKPXQNLMYDNWEERGKPFEGIGGGWGSANX I I II II I I II II I II II II I I I II I II I II II I II orf84ng ATLGGKPQQNLMYDNWEERGKPFEGIGGGWGSANX 370 380 390 Based on this analysis, includng the presence of a putative transmembrane domain (single- underlined) in the gonococcal protein, and a putative ATP/GTP-binding site motif A (P-loop, double-underlined), it is predicted that the proteins from N. meningitidis and N.gonorrhoeae, and their epitopes, could be useful antigens for vaccines or diagnostics, or for raising antibodies. Example 39 The following partial DNA sequence was identified in N. meningitidis <SEQ ID 327>: 1 GTGGTTTTCC TGAATGCCGA CAACGGGATA TTGGTTCAGG ACTTGCCTTT 51 TGAAGTCAAA CTGAAAAAAT TCCATATCGA TTTTTACAAT ACGGGTATGC 101 CGCGTGATTT CGCCAGCGAT ATTGAAGTGA CGGACAAGGC AACCGGTGAG 151 AAACTCGAGC GCACCATCCG CGTGAACCAT CCTTTGACCT TGCACGGCAT 201 CACGATTTAT CAGGCGAGTT TTGCCGACGG CGGTTCGGAT TTGACATTCA 251 AGGCGTGGAA TTTGGGTGAT GCTTCGCGCG AGCCTGTCGT GTTGAAGGCA 301 ACATCCATAC ACCAGTTTCC GTTGGAAATT GGCAAACACA AATATCGTCT 351 TGAGTTCGAT CAGTTCACTT CTATGAATGT GGAGGACATG AGCGAGGGCG 401 CGGAACGGGA AAAAAGCCTG AAATCCACGC TGCCCGATGT CCGCGCCGTT 451 ACTCAGGAAG GTCACAAATA CACCAAT TACCG 501 TATCCGTGAT GCGCCAGGCC AGGCGGTCGA ATATAAAAAC TATATGCTGC 551 CGGTTTTGCA GGAACAGGAT TATTTTTGGA TTACCGGCAC GCGCAGCGC . 601 TTGCAGCAGC AATACCGCTG GCTGCGTATC CCCTTGGACA AGCAGTTGAA 651 AGCGGACACC TTTATGGCAT TGCGTGAGTT TTTGAAAGAT GGGGAAGGGC 701 GCAAACGTCT . GTTGCCGAC GCAACCAAAG GCGCACCTGC CGAAATCCGC 751 GAACAATTCA TGCTGGCTGC GGAAAACACG CTGAACATCT TTGCACAAAA 801 AGGCTATTTG GGATTGGACG AATTTATTAC GTCCAATATC CCGAAAGAGC 851 AGCAGGATAA GATGCAGGGC TATTTCTACG AAATGCTTTA CGGCGTGATG 901 AACGCTGCTT TGGATGAAAC CAT.ACCCGG TACGGCTTGC CCGAATGGCA 951 GCAGGATGAA GCGCGGAATC GTTTCCTGCT GCACAGTATG GATGCGTACA 1001 CGGGTTTGAC CGAATATCCC GCGCCTATGC TGCTGCAACT TGATGGGTTT 1051 TCCGAGGTGC GTTCGTCGGG TTTGCAGATG ACCCGTTCCC C.GGTCCGCT 1101 TTTGGTCTAT CTC... This corresponds to the amino acid sequence <SEQ LD 328; ORF88>: 1 MVFLNADNGI LVQDLPFEVK LKKFHIDFYN TGMPRDFASD IEVTDKATGE 51 KLERTIRVNH PLTLHGITIY QASFADGGSD LTFKAWNLGD ASREPWLKA 101 TSIHQFPLEI GKHKYRLEFD QFTSMNVEDM SEGAEREKSL KSTLPDVRAV 151 TQEGHKYTNX XXXXXYRIRD APGQAVEYKN YMLPVLQEQD YFWITGTRSX 201 LQQQYRWLRI PLDKQLKADT FMALREFLKD GEGRKRXVAD ATKGAPAEIR 251 EQFMLAAENT LNIFAQKGYL GLDEFITSNI PKEQQDKMQG YFYEMLYGVM 301 NAALDETXTR YGLPEWQQDE ARNRFLLHSM DAYTGLTEYP APMLLQLDGF 351 SEVRSSGLQM TRSXGPLLVY L... Further work revealed the complete nucleotide sequence <SEQ ID 329>: 1 ATGAGTAAAT CCCGTAGATC TCCCCCACTT CTTTCCCGTC CGTGGTTCGC 51 TTTTTTCAGC TCCATGCGCT TTGCAGTCGC TTTGCTCAGT CTGCTGGGTA 101 TTGCATCGGT TATCGGTACG GTGTTGCAGC AAAACCAGCC GCAGACGGAT 151 TATTTGGTCA AATTCGGATC GTTTTGGGCG CAGATTTTTG GTTTTCTGGG 201 ACTGTATGAC GTCTATGCTT CGGCATGGTT TGTCGTTATC ATGATGTTTT 251 TGGTGGTTTC TACCAGTTTG TGCCTGATTC GCAATGTGCC GCCGTTCTGG 301 CGCGAAATGA AGTCTTTTCG GGAAAAGGTT AAAGAAAAAT CTCTGGCGGC 351 GATGCGCCAT TCTTCGCTGT TGGATGTAAA AATTGCGCCC GAGGTTGCCA 401 AACGTTATCT GGAAGTACAA GGTTTTCAGG GAAAAACCAT TAACCGTGAA 451 GACGGGTCGG TTCTGATTGC CGCCAAAAAA GGCACAATGA ACAAATGGGG 501 CTATATCTTT GCCCATGTTG CTTTGATTGT CATTTGCCTG GGCGGGTTGA 551 TAGACAGTAA CCTGCTGTTG AAACTGGGTA TGCTGACCGG TCGGATTGTT 601 CCGGACAATC AGGCGGTTTA TGCCAAGGAT TTCAAGCCCG AAAGTATTTT 651 GGGTGCGTCC AATCTCTCAT TTAGGGGCAA CGTCAATATT TCCGAGGGGC 701 AGAGTGCGGA TGTGGTTTTC CTGAATGCCG ACAACGGGAT ATTGGTTCAG 751 GACTTGCCTT TTGAAGTCAA ACTGAAAAAA TTCCATATCG ATTTTTACAA 801 TACGGGTATG CCGCGTGATT TCGCCAGCGA TATTGAAGTG ACGGACAAGG 851 CAACCGGTGA GAAACTCGAG CGCACCATCC GCGTGAACCA TCCTTTGACC 901 TTGCACGGCA TCACGATTTA TCAGGCGAGT TTTGCCGACG GCGGTTCGGA 951 TTTGACATTC AAGGCGTGGA ATTTGGGTGA TGCTTCGCGC GAGCCTGTCG 1001 TGTTGAAGGC AACATCCATA CACCAGTTTC CGTTGGAAAT TGGCAAACAC 1051 AAATATCGTC TTGAGTTCGA TCAGTTCACT TCTATGAATG TGGAGGACAT 1101 GAGCGAGGGC GCGGAACGGG AAAAAAGCCT GAAATCCACG CTGAACGATG 1151 TCCGCGCCGT TACTCAGGAA GGTAAAAAAT ACACCAATAT CGGCCCTTCC 1201 ATTGTTTACC GTATCCGTGA TGCGGCAGGG CAGGCGGTCG AATATAAAAA 1251 CTATATGCTG CCGGTTTTGC AGGAACAGGA TTATTTTTGG ATTACCGGCA 1301 CGCGCAGCGG CTTGCAGCAG CAATACCGCT GGCTGCGTAT CCCCTTGGAC 1351 AAGCAGTTGA AAGCGGACAC CTTTATGGCA TTGCGTGAGT TTTTGAAAGA 1401 TGGGGAAGGG CGCAAACGTC TGGTTGCCGA CGCAACCAAA GGCGCACCTG 1451 CCGAAATCCG CGAACAATTC ATGCTGGCTG CGGAAAACAC GCTGAACATC 1501 TTTGCACAAA AAGGCTATTT GGGATTGGAC GAATTTATTA CGTCCAATAT 1551 CCCGAAAGAG CAGCAGGATA AGATGCAGGG CTATTTCTAC GAAATGCTTT 1601 ACGGCGTGAT GAACGCTGCT TTGGATGAAA CCATACGCCG GTACGGCTTG 1651 CCCGAATGGC AGCAGGATGA AGCGCGGAAT CGTTTCCTGC TGCACAGTAT 1701 GGATGCGTAC ACGGGTTTGA CCGAATATCC CGCGCCTATG CTGCTGCAAC 1751 TTGATGGGTT TTCCGAGGTG CGTTCGTCGG GTTTGCAGAT GACCCGTTCC 1801 CCGGGTGCGC TTTTGGTCTA TCTCGGCTCG GTGCTGTTGG TATTGGGTAC 1851 GGTATTGATG TTTTATGTGC GCGAAAAACG GGCGTGGGTA TTGTTTTCAG 1901 ACGGCAAAAT CCGTTTTGCC ATGTCTTCGG CCCGCAGCGA ACGGGATTTG 1951 CAGAAGGAAT TTCCAAAACA CGTCGAGAGT CTGCAACGGC TCGGCAAGGA 2001 CTTGAATCAT GACTGA This corresponds to the amino acid sequence <SEQ LD 330; ORF88-l>: 1 MSKSRRSPPL LSRPWFAFFS SMRFAVALLS LLGIASVIGT VLQQNQPQTD 51 YLVKFGSFWA QIFGFLGLYD VYASAWFWI MMFLWSTSL CLIRNVPPFW 101 REMKSFREKV KEKSLAAMRH SSLLDVKIAP EVAKRYLEVQ GFQGKTINRE 151 DGSVLIAAKK GTMNKWGYIF AHVALIVICL GGLIDSNLLL KLGMLTGRIV 201 PDNQAVYAKD FKPESILGAS NLSFRGNVNI SEGQSADWF LNADNGILVQ 251 DLPFEVKLKK FHIDFYNTGM PRDFASDIEV TDKATGEKLE RTIRVNHPLT 301 LHGITIYQAS FADGGSDLTF KAWNLGDASR EPWLKATSI HQFPLEIGKH 351 KYRLEFDQFT SMNVEDMSEG AEREKSLKST LNDVRAVTQE GKKYTNIGPS 401 IVYRIRDAAG QAVEYKNYML PVLQEQDYFW ITGTRSGLQQ QYRWLRIPLD 451 KQLKADTFMA LREFLKDGEG RKRLVADATK GAPAEIREQF MLAAENTLNI 501 FAQKGYLGLD EFITSNIPKE QQDKMQGYFY EMLYGVMNAA LDETIRRYGL 551 PEWQQDEARN RFLLHSMDAY TGLTEYPAPM LLQLDGFSEV RSSGLQMTRS 601 PGALLVYLGS VLLVLGTVLM FYVREKRAWV LFSDGKIRFA MSSARSERDL 651 QKEFPKHVES LQRLGKDLNH D* Computer analysis of this amino acid sequence gave the following results: Homology with a predicted ORF from N.meninsitidis (strain A) ORF88 shows 95.7% identity over a 371aa overlap with an ORF (ORF88a) from strain A of N. meningitidis: 10 20 30 orf88.pep MVFLΝADΝGILVQDLPFEVKLKKFHIDFYΝ : I I I I I I I I I II I II I I I I I I I II I I I II I orf88a AKDFKPESILGASΝLSFRGΝVΝISEGQSADWFLΝADΝGILVQDLPFEVKLKKFHIDFYΝ 210 220 230 240 250 260 40 50 60 70 80 90 orf88.pep TGMPRDFASDIEVTDKATGEKLERTIRVΝHPLTLHGITIYQASFADGGSDLTFKAWΝLGD I II I I I I I I I II I II I I I II I I I I I I I I I I I I I II II II I I I II I II I I I II I I I I I I I I orf88a TGMPRDFASDIEVTDKATGEKLERTIRVΝHPLTLHGITIYQASFADGGSDLTFKAWΝLGD 270 280 290 300 310 320 100 110 120 130 140 150 orf88.pep ASREPWLKATSIHQFPLEIGKHKYRLEFDQFTSMΝVEDMSEGAEREKSLKSTLPDVRAV II I I I I I II II I I II I I I II I I II I I I I I I I I I II I I II I I I I I I II I I I I I I I II I I I orf88a ASREPVVLKATSIHQFPLEIGKHKYRLEFDQFTSMΝVEDMSEGAEREKSLKSTLΝDVRAV 330 340 350 360 370 380 160 170 180 190 200 210 orf88.pep TQEGHKYTΝXXXXXXYRIRDAPGQAVEYKΝYMLPVLQEQDYFWITGTRSXLQQQYRWLRI I II I: I I I I I I II II I II I I II I I II I I I II I I II I I I I I I I I II II I I I II orf88a TQEGKKYTΝIGPSIVYRIRDAAGQAVEYKΝYMLPVLQEQDYFWITGTRSGLQQQYRWLRI 390 400 410 420 430 440 220 230 240 250 260 270 orf88.pep PLDKQLKADTFMALREFLKDGEGRKRXVADATKGAPAEIREQFMLAAEΝTLΝIFAQKGYL I I I I I I II I II I II I I I I II I I I I I I I II II I I II II II I II I I I I I I I I II II I I I II orf88a PLDKQLKADTFMALREFLKDGEGRKRLVADATKGAPAEIREQFMLAAEΝTLΝIFAQKGYL 450 460 470 480 490 500 280 290 300 310 320 330 orf88.pep GLDEFITSΝIPKEQQDKMQGYFYEMLYGVMΝAALDETXTRYGLPEWQQDEARΝRFLLHSM I I II I II I I I I II I I I I II I I I II I I I I I I II I I II I II I I I I I I I I I I I II I I I I II orf88a GLDEFITSΝIPKEQQDKMQGYFYEMLYGVMΝAALDETIRRYGLPEWQQDEARΝRFLLHSM 510 520 530 540 550 560 340 350 360 370 orf88.pep DAYTGLTEYPAPMLLQLDGFSEVRSSGLQMTRSXGPLLVYL II I I I I I II I I I I II II I I I II II I I I I I I I I I I I I I II orf88a DAYTGLTEYPAPMLLQLDGFSEVRSSGLQMTRSPGALLVYLGSVLLVLGTVLMFYVREKR 570 580 590 600 610 620 orf88a AWVLFSDGKIRFAMSSARSERDLQKEFPKHVESLQRLGKDLΝHDX 630 640 650 660 670 The complete length ORF88a nucleotide sequence <SEQ LD 331> is: 1 ATGAGTAAAT CCCGTAGATC TCCCCCACTT CTTTCCCGTC CGTGGTTCGC 51 TTTTTTCAGC TCCATGCGCT TTGCGGTCGC TTTGCTCAGT CTGCTGGGTA 101 TTGCATCGGT TATCGGTACG GTGTTGCAGC AAAACCAGCC GCAGACGGAT 151 TATTTGGTCA AATTCGGATC GTTTTGGGCG CAGATTTTTG GTTTTCTGGG 201 ACTGTATGAC GTCTATGCTT CGGCATGGTT TGTCGTTATC ATGATGTTTT 251 TGGTGGTTTC TACCAGTTTG TGCCTGATTC GCAATGTGCC GCCGTTCTGG 301 CGCGAAATGA AGTCTTTTCG GGAAAAGGTT AAAGAAAAAT CTCTGGCGGC 351 GATGCGCCAT TCTTCGCTGT TGGATGTAAA AATTGCGCCC GAGGTTGCCA 401 AACGTTATCT GGAAGTACAA GGTTTTCAGG GAAAAACCAT TAACCGTGAA 451 GACGGGTCGG TTCTGATTGC CGCCAAAAAA GGCACAATGA ACAAATGGGG 501 CTATATCTTT GCCCATGTTG CTTTGATTGT CATTTGCCTG GGCGGGTTGA 551 TAGACAGTAA CCTGCTGTTG AAACTGGGTA TGCTGACCGG TCGGATTGTT 601 CCGGACAATC AGGCGGTTTA TGCCAAGGAT TTCAAGCCCG AAAGTATTTT 651 GGGTGCGTCC AATCTCTCAT TTAGGGGCAA CGTCAATATT TCCGAGGGGC 701 AGAGTGCGGA TGTGGTTTTC CTGAATGCCG ACAACGGGAT ATTGGTTCAG 751 GACTTGCCTT TTGAAGTCAA ACTGAAAAAA TTCCATATCG ATTTTTACAA 801 TACGGGTATG CCGCGCGATT TTGCCAGTGA TATTGAAGTA ACGGATAAGG 851 CAACCGGTGA GAAACTCGAG CGCACCATCC GCGTGAACCA TCCTTTGACC 901 TTGCACGGCA TCACGATTTA TCAGGCGAGT TTTGCCGACG GCGGTTCGGA 951 TTTGACATTC AAGGCGTGGA ATTTGGGTGA TGCTTCGCGC GAGCCTGTCG 1001 TGTTGAAGGC AACATCCATA CACCAGTTTC CGTTGGAAAT TGGCAAACAC 1051 AAATATCGTC TTGAGTTCGA TCAGTTTACT TCTATGAATG TGGAGGACAT 1101 GAGCGAGGGC GCGGAACGGG AAAAAAGCCT GAAATCCACG CTGAACGATG 1151 TCCGCGCCGT TACTCAGGAA GGTAAAAAAT ACACCAATAT CGGCCCTTCC 1201 ATTGTTTACC GTATCCGTGA TGCGGCAGGG CAGGCGGTCG AATATAAAAA 1251 CTATATGCTG CCGGTTTTGC AGGAACAGGA TTATTTTTGG ATTACCGGCA 1301 CGCGCAGCGG CTTGCAGCAG CAATACCGCT GGCTGCGTAT CCCCTTGGAC 1351 AAGCAGTTGA AAGCGGACAC CTTTATGGCA TTGCGTGAGT TTTTGAAAGA 1401 TGGGGAAGGG CGCAAACGTC TGGTTGCCGA CGCAACCAAA GGCGCACCTG 1451 CCGAAATCCG CGAACAATTC ATGCTGGCTG CGGAAAACAC GCTGAACATC 1501 TTTGCACAAA AAGGCTATTT GGGATTGGAC GAATTTATTA CGTCCAATAT 1551 CCCGAAAGAG CAGCAGGATA AGATGCAGGG CTATTTCTAC GAAATGCTTT 1601 ACGGCGTGAT GAACGCTGCT TTGGATGAAA CCATACGCCG GTACGGCTTG 1651 CCCGAATGGC AGCAGGATGA AGCGCGGAAT CGTTTCCTGC TGCACAGTAT 1701 GGATGCGTAC ACGGGTTTGA CCGAATATCC CGCGCCTATG CTGCTGCAAC 1751 TTGATGGGTT TTCCGAGGTG CGTTCGTCGG GTTTGCAGAT GACCCGTTCC 1801 CCGGGTGCGC TTTTGGTCTA TCTCGGCTCG GTGCTGTTGG TATTGGGTAC 1851 GGTATTGATG TTTTATGTGC GCGAAAAACG GGCGTGGGTA TTGTTTTCAG 1901 ACGGCAAAAT CCGTTTTGCC ATGTCTTCGG CCCGCAGCGA ACGGGATTTG 1951 CAGAAGGAAT TTCCAAAACA CGTCGAGAGT CTGCAACGGC TCGGCAAGGA 2001 CTTGAATCAT GACTGA This encodes a protein having amino acid sequence <SEQ LD 332>: 1 MSKSRRSPPL LSRPWFAFFS SMRFAVALLS LLGIASVIGT VLQQNQPQTD 51 YLVKFGSFWA QIFGFLGLYD VYASAWFWI MMFLWSTSL CLIRNVPPFW 101 REMKSFREKV KEKSLAAMRH SSLLDVKIAP EVAKRYLEVQ GFQGKTINRE 151 DGSVLIAAKK GTMNKWGYIF AHVALIVICL GGLIDSNLLL KLGMLTGRIV 201 PDNQAVYAKD FKPESILGAS NLSFRGNVNI SEGQSADWF LNADNGILVQ 251 DLPFEVKLKK FHIDFYNTGM PRDFASDIEV TDKATGEKLE RTIRVNHPLT 301 LHGITIYQAS FADGGSDLTF KAWNLGDASR EPWLKATSI HQFPLEIGKH 351 KYRLEFDQFT SMNVEDMSEG AEREKSLKST LNDVRAVTQE GKKYTNIGPS 401 IVYRIRDAAG QAVEYKNYML PVLQEQDYFW ITGTRSGLQQ QYRWLRIPLD 451 KQLKADTFMA LREFLKDGEG RKRLVADATK GAPAEIREQF MLAAENTLNI 501 FAQKGYLGLD EFITSNIPKE QQDKMQGYFY EMLYGVMNAA LDETIRRYGL 551 PEWQQDEARN RFLLHSMDAY TGLTEYPAPM LLQLDGFSEV RSSGLQMTRS 601 PGALLVYLGS VLLVLGTVLM FYVREKRAWV LFSDGKIRFA MSSARSERDL 651 QKEFPKHVES LQRLGKDLNH D* ORF88a and ORF88-1 100.0% identity in 671 aa overlap: orf88a.pep MSKSRRSPPLLSRPWFAFFSSMRFAVALLSLLGIASVIGTVLQQNQPQTDYLVKFGSFWA 60 I I I II I II I I I I I I I I I I I II I I II I I II I II I I I I II II II II I I I I I I I II I II II II orf88-1 MSKSRRSPPLLSRPWFAFFSSMRFAVALLSLLGIASVIGTVLQQNQPQTDYLVKFGSFWA 60 orf 88a . pep QIFGFLGLYDVYASAWFWIMMFLWSTSLCLIRNVPPFWREMKSFREKVKEKSLAAMRH 120 I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I orf 88-1 QIFGFLGLYDVYASAWFVVIMMFLVVSTSLCLIRNVPPFWREMKSFREKVKEKSLAAMRH 120 orf88a. pep SSLLDVKIAPEVAKRYLEVQGFQGKTINREDGSVLIAAKKGTMNKWGYIFAHVALIVICL 180 I I I II I II I I I I I II I I II I I II I II I I II II I I I I I II II I I I II I I I I I I I I I I I I II orf88-1 SSLLDVKIAPEVAKRYLEVQGFQGKTINREDGSVLIAAKKGTMNKWGYIFAHVALIVICL 180 orf88a. pep GGLIDSNLLLKLGMLTGRIVPDNQAVYAKDFKPESILGASNLSFRGNVNISEGQSADWF 240 II I II II I II I I I I I M I M M I II II II I I II M I I M M I M I I I I I I II II I I I I I I orf88-1 GGLIDSNLLLKLGMLTGRIVPDNQAVYAKDFKPESILGASNLSFRGNVNISEGQSADVVF 240 orf88a. pep LNADNGILVQDLPFEVKLKKFHIDFYNTGMPRDFASDIEVTDKATGEKLERTIRVNHPLT 300 I II II I II I II I II II I II II II I I I I I II II II I I I I II I II II I II I I I II I I II II I orf88-1 LNADNGILVQDLPFEVKLKKFHIDFYNTGMPRDFASDIEVTDKATGEKLERTIRVNHPLT 300 orf88a. pep LHGITIYQASFADGGSDLTFKAWNLGDASREPWLKATSIHQFPLEIGKHKYRLEFDQFT 360 II I II I I I II II I I I I I I I II I II II I I I I I I I II I I I I I I I I I I I I I I I II II I I I I I I orf88-l LHGITIYQASFADGGSDLTFKAWNLGDASREPWLKATSIHQFPLEIGKHKYRLEFDQFT 360 orf88a . pep SMNVEDMSEGAEREKSLKSTLNDVRAVTQEGKKYTNIGPSIVYRIRDAAGQAVEYKNYML 420 II I II I I II I II II II I I I II I II II I I I II II II II II I I I II II I I II I I II I I II I I orf88-1 SMNVEDMSEGAEREKSLKSTLNDVRAVTQEGKKYTNIGPSIVYRIRDAAGQAVEYKNYML 420 orf88a. pep PVLQEQDYFWITGTRSGLQQQYRWLRIPLDKQLKADTFMALREFLKDGEGRKRLVADATK 480 I II II I I II II II II I II II II I I I II I I I II II II I I I I I I I II II I I I I II I I I I I I I orf88-1 PVLQEQDYFWITGTRSGLQQQYRWLRIPLDKQLKADTFMALREFLKDGEGRKRLVADATK 480 orf88a. pep GAPAEIREQFMLAAENTLNIFAQKGYLGLDEFITSNIPKEQQDKMQGYFYEMLYGVMNAA 540 II I II I I I I II II I I I II II II I I I II I I I II II I I I I I I II I II I I I I I II II I I I I I I orf88-1 GAPAEIREQFMLAAENTLNIFAQKGYLGLDEFITSNIPKEQQDKMQGYFYEMLYGVMNAA 540 orf88a. pep LDETIRRYGLPEWQQDEARNRFLLHSMDAYTGLTEYPAPMLLQLDGFSEVRSSGLQMTRS 600 I II I II II I II II II I I II I I II I I I I I II I I I II I I II I I I II II I II I II I II II II I orf88-1 LDETIRRYGLPEWQQDEARNRFLLHSMDAYTGLTEYPAPMLLQLDGFSEVRSSGLQMTRS 600 orf88a . pep PGALLVYLGSVLLVLGTVLMFYVREKRAWVLFSDGKIRFAMSSARSERDLQKEFPKHVES 660 I I II II I II I II I I II I I I II I I II II II II I I I II I I I I I II II II I I I I II I I I I I I I orf88-l PGALLVYLGSVLLVLGTVLMFYVREKRAWVLFSDGKIRFAMSSARSERDLQKEFPKHVES 660 orf88a. pep LQRLGKDLNHD 672 I I II I II I I I I orf88-l LQRLGKDLNHD 672 Homology with a predicted ORF from N.sonorrhoeae ORF88 shows 93.8% identity over a 371aa overlap with a predicted ORF (ORF88.ng) from N. gonorrhoeae: orf 88 . pep MVFLNADNGILVQDLPFEVKLKKFHIDFYNTGMPRDFASDIEVTDKATGEKLERTIRVNH 60 I I M M I I I : M I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I orf 88ng MVFLNADNGMLVQDLPFEVKLKKFHIDFYNTGMPRDFASDIEVTDKATGEKLERTIRVNH 60 orf 88 . pep PLTLHGITIYQASFADGGSDLTFKAWNLGDASREPWLKATΞIHQFPLEIGKHKYRLEFD 120 I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I orf88ng PLTLHGITIYQASFADGGSDLTFKAWNLRDASREPWLKATSIHQFPLEIGKHKYRLEFD 120 orf 88 . pep QFTSMNVEDMSEGAEREKSLKSTLPDVRAVTQEGHKYTNXXXXXXYRIRDAPGQAVEYKN 180 II I I I I I I I I I II I I I I I I I II I I I I I I I I I MM I I I I I I I I I I I I II II I orf88ng QFTSMNVEDMSEGAEREKSLKSTLNDVRAVTQEGKKYTNIGPSIVYRIRDAAGQAVEYKN 180 orf88.pep YMLPVLQEQDYFWITGTRSXLQQQYRWLRIPLDKQLKADTFMALREFLKDGEGRKRXVAD 240 I I II: I M: I II I : I I I II I I II II I II I I I II I I I I I I I I I II I I I II I I I II I III orf88ng YMLPILQDKDYFWLTGTRSGLQQQYRWLRIPLDKQLKADTFMALREFLKDGEGRKRLVAD 240 orf88. pep ATKGAPAEIREQFMLAAENTLNIFAQKGYLGLDEFITSNIPKEQQDKMQGYFYEMLYGVM 300 III I I I I I II I I I I I I I II II I I I I I I I II II I I I II II I I I II I I I I I I II I I I I I I orf88ng ATKDAPAEIREQFMLAAENTLNIFAQKGYLGLDEFITSNIPKGQQDKMQGYFYEMLYGVM 300 orf88.pep NAALDETXTRYGLPEWQQDEARNRFLLHSMDAYTGLTEYPAPMLLQLDGFSEVRSSGLQM 360 I I I I I II I I I I II I I I I I II I I I I I I II I I I I I I I I I II I II II II I I I II I I I I I I I orf88ng NAALDETIRRYGLPEWQQDEARNRFLLHSMDAYTGLTEYPAPMLLQLDGFSEVRSSGLQM 360 orf88. pep TRSXGPLLVYL 371 III I I I I II orf88ng TRSPGALLVYLGSVLLVLGTVFMFYVPKKRAWVLFSNXKIRFAMSSARSERDLQKEFPKH 420 An ORF88ng nucleotide sequence <SEQ ED 333> was predicted to encode a protein having amino acid sequence <SEQ LD 334>: 1 MVFLNADNGM LVQDLPFEVK LKKFHIDFYN TGMPRDFASD IEVTDKATGE 51 KLERTIRVNH PLTLHGITIY QASFADGGSD LTFKAWNLRD ASREPWLKA 101 TSIHQFPLEI GKHKYRLEFD QFTSMNVEDM SEGAEREKSL KSTLNDVRAV 151 TQEGKKYTNI GPSIVYRIRD AAGQAVEYKN YMLPILQDKD YFWLTGTRSG 201 LQQQYRWLRI PLDKQLKADT FMALREFLKD GEGRKRLVAD ATKDAPAEIR 251 EQFMLAAENT LNIFAQKGYL GLDEFITSNI PKGQQDKMQG YFYEMLYGVM 301 NAALDETIRR YGLPEWQQDE ARNRFLLHSM DAYTGLTEYP APMLLQLDGF 351 SEVRSSGLQM TRSPGALLVY LGSVLLVLGT VFMFYVPKKR AWVLFSNXKI 401 RFAMSSARSE RDLQKEFPKH VESLQRLGKD LNHD* Further work revealed the complete gonococcal DNA sequence <SEQ LD 335>: 1 ATGAGTAAAT CCCGTATATC TCCCACACTT CTTTCCCGTC CGTGGTTCGC 51 TTTTTTCAGC TCCATGCGCT TTGCGGTCGC TTTGCTCAGT CTGCTGGGTA 101 TTGCATCGGT TATCGGCACG GTGTTACAGC AAAACCAGCC GCAGACGGAT 151 TATTTGGTCA AATTCGGACC GTTTTGGACT CGGATTTTTG ATTTTTTGGG 201 TTTGTATGAT GTCTATGCTT CGGCATGGTT TGTCGTTATC ATGATGTTTC 251 TGGTGGTTTC TACCAGTTTG TGTTTAATCC GTAACGTTCC GCCGTTTTGG 301 CGCGAAATGA AGTCTTTCCG GGAAAAGGTT AAAGAAAAAT CTCTGGCGGC 351 GATGCGCCAT TCTTCGCTGT TGGATGTAAA AATTGCCCCC GAAGTTGCCA 401 AACGTTATCT GGAGGTGCGG GGTTTTCAGG GAAAAACCGT CAGCCGTGAG 451 GACGGGTCGG TTCTGATTGC CGCCAAAAAA GGCAcaatga acaaATGGGG 501 CTATATCTTT GCccaagtag ctTTGATTGT CATTTGCCTG GGCGGGTTGA 551 TAGACAGTAA CCTGCTGCTG AAGCTGGGTA TGCTGGCCGG TCGGATTGTT 601 CCGGACAATC AGGCGGTTTA TGCCAAGGAT TTCAAGCCCG AAAGTATTTT 651 GGGTGCGTCC AATCTCTCAT TTAGGGGCAA CGTCAATATT TCCGAGGGGC 701 AAAGTGCGGA TGTGGTTTTC CTGAATGCCG ACAACGGGAT GTTGGTTCAG 751 GACTTGCCTT TTGAAGTCAA ACTGAAAAAA TTCCATATCG ATTTTTACAA 801 TACGGGTATG CCGCGCGATT TTGCCAGCGA TATTGAAGTA ACGGACAAGG 851 CAACCGGTGA GAAACTCGAG CGCACCATCC GCGTGAACCA TCCTTTGACC 901 TTGCACGGCA TCACGATTTA TCAGGCGAGT TTTGCCGACG GCGGTTCGGA 951 TTTGACATTC AAGGCGTGGA ATTTGAGGGA TGCTTCGCGC GAACCTGTCG 1001 TGTTGAAGGC AACCTCCATA CACCAGTTTC CGTTGGAAAT CGGCAAACAC 1051 AAATATCGTC TTGAGTTCGA TCAGTTCACT TCTATGAATG TGGAGGACAT 1101 GAGCGAGGGT GCGGAACGGG AAAAAAGCCT GAAATCCACT CTGAACGATG 1151 TCCGCGCCGT TACTCAGGAA GGTAAAAAAT ACACCAATAT CGGCCCTTCC 1201 ATCGTGTACC GCATCCGTGA TGcggCAGGG CAGGCGGTCG AATATAAAAA 1251 CTATATGCTG CCGATTTTGC AGGACAAAGA TTATTTTTGG CTGACCGGCA 1301 CGCGCAGCGG CTTGCAGCAG CAATACCGCT GGCTGCGTAT CCCCTTGGAC 1351 AAGCAGTTGA AAGCGGACAC CTTTATGGCA TTGCGTGAGT TTTTGAAAGA 1401 TGGGGAAGGG CGCAAACGTC TGGTTGCCGA CGCAACCAAA GACGCACCTG 1451 CCGAAATCCG CGAACAATTC ATGCTGGCTG CGGAAAACAC GCTGAATATC 1501 TTTGCGCAAA AAGGCTATTT GGGATTGGAC GAATTTATTA CGTCCAATAT 1551 CCCGAAAGGG CAGCAGGATA AGATGCAGGG CTATTTCTAC GAAATGCTTT 1601 ACGGCGTGAT GAACGCTGCT TTGGATGAAA CCATACGCCG GTACGGCTTG 1651 CCCGAATGGC AGCAGGATGA AGCGCGGAAC CGTTTCCTGC TGCACAGTAT 1701 GGATGCCTAT ACGGGGCTGA CGGAATATCC CGCGCCTATG CTGCTCCAGC 1751 TTGACGGGTT TTCCGAGGTG CGTTCCTCAG GTTTGCAGAT GACCCGTTCG 1801 CCGGGTGCGC TTTTGGTCTA TCtcggctcg gtattgttgg TTTTGGgtac 1851 ggtaTttatg tTTTATGTGC GCGAAAAACG GGCGTGGgta tTGTTTTCag 1901 aCGGCAAAAT CCGTTTTGCT ATGtCTTcgg CCcgcagcga ACGGGATTTG 1951 cAGAaggaaT TTCCAAAACA CGtcgAGAGC CTGCAACggc tcggcaaggA 2001 CttgaaTCAT GACTga This corresponds to the amino acid sequence <SEQ ID 336; ORF88ng-l>: 1 MSKSRISPTL LSRPWFAFFS SMRFAVALLS LLGIASVIGT VLQQNQPQTD 51 YLVKFGPFWT RIFDFLGLYD VYASAWFWI MMFLWSTSL CLIRNVPPFW 101 REMKSFREKV KEKSLAAMRH SSLLDVKIAP EVAKRYLEVR GFQGKTVSRE 151 DGSVLIAAKK GTMNKWGYIF AQVALIVICL GGLIDSNLLL KLGMLAGRIV 201 PDNQAVYAKD FKPESILGAS NLSFRGNVNI SEGQSADVVF LNADNGMLVQ 251 DLPFEVKLKK FHIDFYNTGM PRDFASDIEV TDKATGEKLE RTIRVNHPLT 301 LHGITIYQAS FADGGSDLTF KAWNLRDASR EPWLKATSI HQFPLEIGKH 351 KYRLEFDQFT SMNVEDMSEG AEREKSLKST LNDVRAVTQE GKKYTNIGPS 401 IVYRIRDAAG QAVEYKNYML PILQDKDYFW LTGTRSGLQQ QYRWLRIPLD 451 KQLKADTFMA LREFLKDGEG RKRLVADATK DAPAEIREQF MLAAENTLNI 501 FAQKGYLGLD EFITSNIPKG QQDKMQGYFY EMLYGVMNAA LDETIRRYGL 551 PEWQQDEARN RFLLHSMDAY TGLTEYPAPM LLQLDGFSEV RSSGLQMTRS 601 PGALLVYLGS VLLVLGTVFM FYVREKRAWV LFSDGKIRFA MSSARSERDL 651 QKEFPKHVES LQRLGKDLNH D* ORF88ng-l and ORF88-1 show 97.0% identity in 671 aa overlap: orf88-l.pep MSKSRRS PPLLSRPWFAFFS SMRFAVALLS LLGIASVIGTVLQQNQPQT DYLVKFGS FWA 60 I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I M orf88ng-l MSKSRISPTLLSRPWFAFFSSMRFAVALLSLLGIASVIGTVLQQNQPQTDYLVKFGPFWT 60 orf88-1. pep QIFGFLGLYDVYASAWFWIMMFLWSTSLCLIRNVPPFWREMKSFREKVKEKSLAAMRH 120 Ml I I I II II II I I I I I I I II I I II I I I II II I II I I I I I I II II I I I I I II I II I I II orf88ng- l RIFDFLGLYDVYASAWFWIMMFLWSTSLCLIRNVPPFWREMKSFREKVKEKSLAAMRH 120 orf88-1.pep SSLLDVKIAPEVAKRYLEVQGFQGKTINREDGSVLIAAKKGTMNKWGYIFAHVALIVICL 180 I I II I I II I I II II I I I II: I I II I M: II I I I II I II II I I I I I I I I I I I : I II I I I I I orf88ng-l SSLLDVKIAPEVAKRYLEVRGFQGKTVSREDGSVLIAAKKGTMNKWGYIFAQVALIVICL 180 orf88-1.pep GGLIDSNLLLKLGMLTGRIVPDNQAVYAKDFKPESILGASNLSFRGNVNISEGQSADWF 240 II I I I I I II II II II : II I I I I I I II I I I I I II I I II II II II II II I II I I I I I I I I I I orf88ng-l GGLIDSNLLLKLGMLAGRIVPDNQAVYAKDFKPESILGASNLSFRGNVNISEGQSADWF 240 orf88-1. pep LNADNGILVQDLPFEVKLKKFHIDFYNTGMPRDFASDIEVTDKATGEKLERTIRVNHPLT 300 II I II I : I I I I I I I II I II II I II I I II II I I I II I II I I I I II II I I I I II II I II I II orf88ng-l LNADNGMLVQDLPFEVKLKKFHIDFYNTGMPRDFASDIEVTDKATGEKLERTIRVNHPLT 300 orf88-l.pep LHGITIYQASFADGGSDLTFKAWNLGDASREPWLKATSIHQFPLEIGKHKYRLEFDQFT 360 I I II I I II II I II I I I II II I II II II II II II I I I I II I I I I I I I I I I I II I II II II orf88ng-l LHGITIYQASFADGGSDLTFKAWNLRDASREPWLKATSIHQFPLEIGKHKYRLEFDQFT 360 orf88-1.pep SMNVEDMSEGAEREKSLKSTLNDVRAVTQEGKKYTNIGPSIVYRIRDAAGQAVEYKNYML 420 I I II I I I II II I I I I I I II II I II I I II I II II I II I I I I I I II I II I II I I I II I I I I I orf88ng-l SMNVEDMSEGAEREKSLKSTLNDVRAVTQEGKKYTNIGPSIVYRIRDAAGQAVEYKNYML 420 orf88-1.pep PVLQEQDYFWITGTRSGLQQQYRWLRIPLDKQLKADTFMALREFLKDGEGRKRLVADATK 480 I : I I :: I I I I : I II I I I I I I I I I II II I I I I I I II I I I II I I I I I I I I I I II II I II I I I orf88ng-l PILQDKDYFWLTGTRSGLQQQYRWLRIPLDKQLKADTFMALREFLKDGEGRKRLVADATK 480 orf88-1. pep GAPAEIREQFMLAAENTLNIFAQKGYLGLDEFITSNIPKEQQDKMQGYFYEMLYGVMNAA 540 II I I I II II I I II I I II II I II I I I I I I II II I I II II I I I II II I II I II II I I I II orf88ng-l DAPAEIREQFMLAAENTLNIFAQKGYLGLDEFITSNIPKGQQDKMQGYFYEMLYGVMNAA 540 orf88-1. pep LDETIRRYGLPEWQQDEARNRFLLHSMDAYTGLTEYPAPMLLQLDGFSEVRSSGLQMTRS 600 I I I I I II II II I I I II I I I I II I I I II I II II I II I I I II II II II I I I I I I I I I I I I I I orf88ng-l LDETIRRYGLPEWQQDEARNRFLLHSMDAYTGLTEYPAPMLLQLDGFSEVRSSGLQMTRS 600 orf88-1. pep PGALLVYLGSVLLVLGTVLMFYVREKRAWVLFSDGKIRFAMSSARSERDLQKEFPKHVES 660 I I I II I I I I I II II I I II : I I II I I I II I II II I I II I I I I II II II I II I II I II I II I orf88ng-l PGALLVYLGSVLLVLGTVFMFYVREKRAWVLFSDGKIRFAMSSARSERDLQKEFPKHVES 660 orf88-1. pep LQRLGKDLNHD 671 II I I I I II II I orf88ng-l LQRLGKDLNHD 671 Furthermore, ORG88ng-l shows homology with a hypothetical protein from Aquifex aeolicus: gi 12984296 (AE000771) hypothetical protein [Aquifex aeolicus] Length = 537 Score = 94.4 bits (231), Expect = 2e-18 Identities = 91/334 (27%), Positives = 159/334 (47%), Gaps = 59/334 (17%) Query: 16 FAFFSSMRFAVALLSLLGIASVIG-TVLQQNQPQTDYLVKFGPFWTRIFDFLGLYDVYAS 74 + F +S++ A+ ++ +LGI S++G T ++QNQ YL +FG L L DV+ S Sbjct: 80 YDFLASLKLAIFIMLVLGILSMLGSTYIKQNQSFEWYLDQFGYDVGIWIWKLWLNDVFHS 139 Query : 75 AWFWIMMFLWSTSLCLIRNVPPFWREMKSFREKVKEKSLAAMRHSSLLDVKIAPEVAK 134 ++++ ++ L V+ C 1+ +P W++ S +E++ + A +H + VKI P+ K Sbj ct : 140 WYYILFIVLLAVNLIFCSIKRLPRVWKQAFS-KERILKLDEHAEKHLKPITVKI-PDKDK 197 Query : 135 — RYLEVRGFQGKTVSREDGSVLIAAKKGTMNKWGYIFAQVALIVICLGGLIDSNLLLKL 192 ++L +GF+ V E + + A+KG ++ G +AL+VI G LID Sbj ct : 198 VLKFLLKKGFK-VFVEEEGNKLYVFAEKGRFSRLGVYITHIALLVIMAGALID 249 Query: 193 GMLAGRIVPDNQAVYAKDFKPESILGASNLSFRGNVNISEGQSADWFLNADNGMLVQDL 252 +I+G RG++ ++EG + DV+ + A+ L Sbjct: 250 AIVGV RGSLIVAEGDTNDVMLVGAE—QKPYKL 280 Query: 253 PFEVKLKKFHIDFY NTGMPRDFA SDIEVTDKATGEKLER—TIRVNHPLT 300 PF V L F I Y N + + FA SDIE+ + G K+E T++VN P Sbjct: 281 PFAVHLIDFRIKTYAEENPNVDKRFAQAVSSYESDIEIIN GGKVEAKGTVKVNEPFD 337 Query: 301 LHGITIYQASFA—DGGSDLTFKAWNLRDASREP 332 ++QA++ DG S + + + A +P Sbjct: 338 FGRYRLFQATYGILDGTSGMGVIWDRKKAHEDP 371 Based on this analysis, including the putative transmembrane domain in the gonococcal protein, it is predicted that the proteins from N. meningitidis and N.gonorrhoeae, and their epitopes, could be useful .antigens for vaccines or diagnostics, or for raising antibodies. Example 40 The following DNA sequence, believed to be complete, was identified in N. meningitidis <SEQ LD 337>: 1 ATGATGAGTA ATAmAATGGm ACAAAAAGGG TTTACATTGA TTGmGmTGAT 51 GATAGTCGTC GCGATACTCG GCATTATCAG CGTCATTGCC ATACCTTCTT 101 ATCmAAGTTA TATTGAAAAA GGCTATCAGT CCCAGCTTTA TACGGAGATG 151 GyCGGTATCA ACAATATTTC CAAACAGTTT ATTTTGAAAA ATCCCCTGGA 201 CGATAATCAG ACCATCGAGA ACAAACTGGA AATATTTGTC TCAGGCTATA 251 AGATGAATCC GAAAATTGCC AAAAAaTATA GTGTTTCGGT AAAGTTTGTC 301 GATAAGGAAA AATCAAGGGC ATACAGGTTG GTCGGCGTTC CGAAGGCGGG 351 GACGGGTTAT ACTTTGTCGG TATGGATGAA CAGCGTGGGC GACGGATACA 401 AATGCCGTGA TGCCGCTTCT GCCCAAGCCC ATTTGGAGAC CTTGTCCTCA 451 GATGTCGGCT GTGAAGCCTT CTCTAATCGT AAAAAATAA This corresponds to the amino acid sequence <SEQ ID 338; ORF89>: 1 MMSNXMXQKG FTLIXXMIW AILGIISVIA IPSYXSYIEK GYQSQLYTEM 51 XGINNISKQF ILKNPLDDNQ TIENKLEIFV SGYKMNPKIA KKYSVSVKFV 101 DKEKSRAYRL VGVPKAGTGY TLSVWMNSVG DGYKCRDAAS AQAHLETLSS 151 DVGCEAFSNR KK* Further work revealed the complete nucleotide sequence <SEQ ID 339>: 1 ATGATGAGTA ATAAAATGGA ACAAAAAGGG TTTACATTGA TTGAGATGAT 51 GATAGTCGTC GCGATACTCG GCATTATCAG CGTCATTGCC ATACCTTCTT 101 ATCAAAGTTA TATTGAAAAA GGCTATCAGT CCCAGCTTTA TACGGAGATG 151 GTCGGTATCA ACAATATTTC CAAACAGTTT ATTTTGAAAA ATCCCCTGGA 201 CGATAATCAG ACCATCGAGA ACAAACTGGA AATATTTGTC TCAGGCTATA 251 AGATGAATCC GAAAATTGCC AAAAAATATA GTGTTTCGGT AAAGTTTGTC 301 GATAAGGAAA AATCAAGGGC ATACAGGTTG GTCGGCGTTC CGAAGGCGGG 351 GACGGGTTAT ACTTTGTCGG TATGGATGAA CAGCGTGGGC GACGGATACA 401 AATGCCGTGA TGCCGCTTCT GCCCAAGCCC ATTTGGAGAC CTTGTCCTCA 451 GATGTCGGCT GTGAAGCCTT CTCTAATCGT AAAAAATAA This corresponds to the amino acid sequence <SEQ ID 340; ORF89-l>: 1 MMSNKMEQKG FTLIEMMIW AILGIISVIA IPSYQSYIEK GYQSQLYTEM 51 VGINNISKQF ILKNPLDDNQ TIENKLEIFV SGYKMNPKIA KKYSVSVKFV 101 DKEKSRAYRL VGVPKAGTGY TLSVWMNSVG DGYKCRDAAS AQAHLETLSS 151 DVGCEAFSNR KK* Computer analysis of this amino acid sequence gave the following results: Homology with PilE of N sonorrhoeae (accession number Z692601. ORF89 and PilE protein show 30% aa identity in 120a overlap: orf89 8 QKGFTLIXXMIWAILGIISVIAIPSYXSYIEKGYQSQLYTEMXGINNISKQFILKNPL- 66 QKGFTLI MIV+AI+GI++ +A+P+Y Y + S+ G + ++ L + + PilE 5 QKGFTLIELMIVIAIVGILAAVALPAYQDYTARAQVSEAILLAEGQKSAVTEYYLNHGIW 64 orf89 67 -DDNQTIENKLEIFVΞGYKMNPKIAKKYSVSVKFVDKEKSRAYRLVGVPKAGTGYTLSVW 125 DN + +G + KI KY SV + GV K G LS+W PilE 65 PKDNTS AGVASSDKIKGKYVQSVTVAKGWTAEMASTGVNKEIQGKKLSLW 115 Homology with a predicted ORF from N.meninsitidis (strain A) ORF89 shows 83.3% identity over a 162aa overlap with an ORF (ORF89a) from strain A of N. meningitidis: 10 20 30 40 50 60 orf 89 . pep MMSNXMXQKGFTLIXXMIWAILGIISVIAIPSYXSYIEKGYQSQLYTEMXGINNISKQF I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I orf 89a MMSNKMEQKGFTLIXXXXXXAIXXXXSVIXXXXYXSYIEKGYQSQLYTEMVGINNISKQX 10 20 30 40 50 60 70 80 90 100 110 120 orf89. pep ILKNPLDDNQTIENKLEIFVSGYKMNPKIAKKYSVSVKFVDKEKSRAYRLVGVPKAGTGY II I II II I I I I I :: II I I II I I I I I I II I I : I I : I II : I I :: II III II II I I : II I I orf89a ILKNPLDDNQTIKSKLEIFVSGYKMNPKIAEKYNVSVHFVNEEKPRAYSLVGVPKTGTGY 70 80 90 100 110 120 130 140 150 160 orf89.pep TLSVWMNSVGDGYKCRDAASAQAHLETLSSDVGCEAFSNRKKX I II I II I I II II I I I I I II II : I I II I I I I II I II I II I I II I orf89a TLSVWMNSVGDGYKCRDAASARAHLETLSSDVGCEAFSNRKKX 130 140 150 160 The complete length ORF89a nucleotide sequence <SEQ ID 341> is: 1 ATGATGAGTA ATAAAATGGA ACAAAAAGGG TTTACATTGA TTGNGANGNT 51 NATNGNCNTC GCGATACNCN GCNTTANCAG CGTCATTNCN ATNNNTNCNT 101 ATCNNAGTTA TATTGAAAAA GGCTATCAGT CCCAGCTTTA TACGGAGATG 151 GTCGGTATCA ACAATATTTC CAAACAGTNT ATTTTGAAAA ATCCCCTGGA 201 CGATAATCAG ACCATCAAGA GCAAACTGGA AATATTTGTC TCAGGCTATA 251 AGATGAATCC GAAAATTGCC GAAAAATATA ATGTTTCGGT GCATTTTGTC 301 AATGAGGAAA AACCNAGGGC ATACAGCTTG GTCGGCGTTC CAAAGACGGG 351 GACGGGTTAT ACTTTGTCGG TATGGATGAA CAGCGTGGGC GACGGATACA 401 AATGCCGTGA TGCCGCTTCT GCCCGAGCCC ATTTGGAGAC CTTGTCCTCA 451 GATGTCGGCT GTGAAGCCTT CTCTAATCGT AAAAAATAG This encodes a protein having amino acid sequence <SEQ LD 342>: 1 MMSNKMEQKG FTLIXXXXXX AIXXXXSVIX XXXYXSYIEK GYQSQLYTEM 51 VGINNISKQX ILKNPLDDNQ TIKSKLEIFV SGYKMNPKIA EKYNVSVHFV 101 NEEKPRAYSL VGVPKTGTGY TLSVWMNSVG DGYKCRDAAS ARAHLETLSS 151 DVGCEAFSNR KK* ORF89a and ORF89-1 show 83.3% identity in 162 aa overlap: 10 20 30 40 50 60 orf89a.pep MMSNKMEQKGFTLIXXXXXXAIXXXXSVIXXXXYXSYIEKGYQSQLYTEMVGINNISKQX I II I II II II II II II III I I II I I I I I II II I I I I II II II I I orf89-1 MMSNKMEQKGFTLIEMMIWAILGIISVIAIPSYQSYIEKGYQSQLYTEMVGINNISKQF 10 20 30 40 50 60 70 80 90 100 110 120 orf89a. pep ILKNPLDDNQTIKSKLEIFVSGYKMNPKIAEKYNVSVHFVNEEKPRAYSLVGVPKTGTGY I II I I I II I I II :: I I I I II I I I II I I II I : II : II I : I I :: II III I I II I I : I II I orf89-l ILKNPLDDNQTIENKLEIFVSGYKMNPKIAKKYSVSVKFVDKEKSRAYRLVGVPKAGTGY 70 80 90 100 110 120 130 140 150 160 orf89a. pep TLSVWMNSVGDGYKCRDAASARAHLETLSSDVGCEAFSNRKKX I I I I II II I I I I II I I II II I M II I I II I II I I II I II I I I I orf89-1 TLSVWMNSVGDGYKCRDAASAQAHLETLSSDVGCEAFSNRKKX 130 140 150 160 Homology with a predicted ORF from Nsonorrhoeae ORF89 shows 84.6% identity over a 162aa overlap with a predicted ORF (ORF89.ng) from N. gonorrhoeae: orf89 MMSNXMXQKGFTLIXXMIWAILGIISVIAIPSYXSYIEKGYQSQLYTEMXGINNISKQF 60 II I I I I I I I II I II I I : I II I I I I II II II I I I I I I I I I I I I II I IMM III orf89ng MMSNKMEQKGFTLIEMMIWTILGIISVIAIPSYQSYIEKGYQSQLYTEMVGINNVLKQF 60 orf89 ILKNPLDDNQTIENKLEIFVSGYKMNPKIAKKYSVSVKFVDKEKSRAYRLVGVPKAGTGY 120 I I I I I II I : I ::: II : I I I II I II II II I I II I II I : II I II I I I I I II II : I I I I I orf89ng ILKNPQDDNDTLKSKLKIFVSGYKMNPKIAKKYSVSVRFVDAEKPRAYRLVGVPNAGTGY 120 orf89 TLSVWMNSVGDGYKCRDAASAQAHLETLSSDVGCEAFSNRKK 162 II I I II I I I II I I II I I I: I II M : I II: I I I II II I I II orf89ng TLSVWMNSVGDGYKCRDATSAQAYSDTLSADSGCEAFSNRKK 162 The complete length ORF89ng nucleotide sequence <SEQ ID 343> is: 1 aTGATGAGCA ATAAAATGGA ACAAAAAGGG TTTACATTGA TTGAGATGAT 51 GATAGTTGTC ACGATACTCG GCATCATCAG CGTCATTGCC ATACCTTCTT 101 ATCAGAGTTA TATTGAAAAA GGCTATCAGT CCCAGCTTTA TACGGAGATG 151 GTCGGTATCA ACAATGTTCT CAAACAGTTT ATTTTGAAAA ATCCCCAGGA 201 CGATAATGAT ACCCTCAAGA GCAAACTGAA AATATTTGTC TCAGGCTATA 251 AGATGAATCC GAAAAttgCC AAAAAATATA GTGTTTCGGt aaggtttGTC 301 gatGCGGAAA AACCAAGGGC ATACAGGTTG GTCGGCGTTC CGAACGCGGG 351 GACGGGTTAT ACTTTGTCGG TATGGATGAA CAGCGTGGGC GACGGATACA 401 AATGCCGTGA TGCCACTTCT GCCCAGGCCT ATTCGGACAC CTTGTCCGCA 451 GATAGCGGCT GTGAAGCTTT CTCTAATCGT AAAAAATAG This encodes a protein having amino acid sequence <SEQ ID 344>: 1 MMSNKMEQKG FTLIEMMIW TILGIISVIA IPSYQSYIEK GYQSQLYTEM 51 VGINNVLKQF ILKNPQDDND TLKSKLKIFV SGYKMNPKIA KKYSVSVRFV 101 DAEKPRAYRL VGVPNAGTGY TLSVWMNSVG DGYKCRDATS AQAYSDTLSA 151 DSGCEAFSNR KK* This gonococcal protein has a putative leader peptide (underlined) and N-terminal methylation site (NMePhe or type-4 pili, double-underlined). In addition, ORF89ng and ORF89-1 show 88.3% identity in 162 aa overlap: 10 20 30 40 50 60 orf89-1. pep MMSNKMEQKGFTLIEMMIWAILGIISVIAIPSYQSYIEKGYQSQLYTEMVGINNISKQF I I I I I I I II I II I II II I I M II I II I II II I I I I I II II I I I I I I I I II I II I I: III orf89ng MMSNKMEQKGFTLIEMMIWTILGIISVIAIPSYQSYIEKGYQSQLYTEMVGINNVLKQF 10 20 30 40 50 60 70 80 90 100 110 120 orf89-1.pep ILKNPLDDNQTIENKLEIFVSGYKMNPKIAKKYSVSVKFVDKEKSRAYRLVGVPKAGTGY II II I II I: I ::: I M I I II I II II I II I I I II I I I : I I I II I II I II I I IM M II orf89ng ILKNPQDDNDTLKSKLKIFVSGYKMNPKIAKKYSVSVRFVDAEKPRAYRLVGVPNAGTGY 70 80 90 100 110 120 130 140 150 160 orf89-1.pep TLSVWMNSVGDGYKCRDAASAQAHLETLSSDVGCEAFSNRKKX I I I I I I I II I I I I I II II : I I II : : II I : I I I II I I II II I orf89ng TLSVWMNSVGDGYKCRDATSAQAYSDTLSADSGCEAFSNRKKX 130 140 150 160 Based on this analysis, including the gonococcal motifs and the homology with the known PilE protein, it was predicted that these proteins from N. meningitidis and N.gonorrhoeae, and their epitopes, could be useful .antigens for vaccines or diagnostics, or for raising antibodies. ORF89-1 (13.6kDa) was cloned in the pGex vector and expressed in E.coli, as described above. The products of protein expression and purification were analyzed by SDS-PAGE. Figure 11A shows the results of affinity purification of the GST- fusion protein. Purified GST-fusion protein was used to immunise mice, whose sera gave a positive result in the ELISA test., confirming that ORF89-1 is a surface-exposed protein, and that it is a useful immunogen. Example 41 The following partial DNA sequence was identified in N. meningitidis <SEQ ID 345>: 1 ATGAAAAAAT CCTCCCTCAT CAGCGCATTG GGCATCGGTA TTTTGAGCAT 51 CGGCATGGCA TTTGCCGCCC CTGCCGACGC GGTAAGCCAA ATCCGTCAAA 101 ACGCCACTCA AGTATTGAGC ATCTTAAAAA ACGGCGATGC CAACACCGCT 151 CGCCAAAAAG CCGAAGCCTA TGCGATTCCC TATTTCGATT TCCAACGTAT 201 GACCGCATTG GCGGTCGGCA ACCCTTGGsG CACCG.GTCC GACG.GCAAA 251 AACAAGCGTT GGCCn.AGAA TTTCAACCC... This corresponds to the amino acid sequence <SEQ ID 346; ORF91>: 1 MKKSSLISAL GIGILSIGMA FAAPADAVSQ IRQNATQVLS ILKNGDANTA 51 RQKAEAYAIP YFDFQRMTAL AVGNPWXTXS DXQKQALAXE FQP ... Further work revealed the complete nucleotide sequence <SEQ ID 347>: 1 ATGAAAAAAT CCTCCCTCAT CAGCGCATTG GGCATCGGTA TTTTGAGCAT 51 CGGCATGGCA TTTGCCGCCC CTGCCGACGC GGTAAGCCAA ATCCGTCAAA 101 ACGCCACTCA AGTATTGAGC ATCTTAAAAA ACGGCGATGC CAACACCGCT 151 CGCCAAAAAG CCGAAGCCTA TGCGATTCCC TATTTCGATT TCCAACGTAT 201 GACCGCATTG GCGGTCGGCA ACCCTTGGCG CACCGCGTCC GACGCGCAAA 251 AACAAGCGTT GGCCAAAGAA TTTCAAACCC TGCTGATCCG CACCTATTCC 301 GGCACGATGC TGAAATTAAA AAACGCCAAC GTCAACGTCA AAGACAATCC 351 CATCGTCAAT AAAGGCGGCA AAGAAATCAT CGTCCGCGCC GAAGTCGGCG 401 TACCCGGGCA AAAACCCGTC AACATGGACT TCACCACCTA CCAAAGCGGC 451 GGTAAATACC GTACCTACAA CGTCGCCATC GAAGGCGCGA GCCTGGTTAC 501 CGTGTACCGC AACCAATTCG GCGAAATTAT CAAAGCGAAA GGCGTGGACG 551 GACTGATTGC CGAGTTGAAA GCCAAAAACG GCGGCAAATA A This corresponds to the amino acid sequence <SEQ ID 348; ORF91-l>: 1 MKKSSLISAL GIGILSIGMA FAAPADAVSQ IRQNATQVLS ILKNGDANTA 51 RQKAEAYAIP YFDFQRMTAL AVGNPWRTAS DAQKQALAKE FQTLLIRTYS 101 GTMLKLKNAN VNVKDNPIVN KGGKEIIVRA EVGVPGQKPV NMDFTTYQSG 151 GKYRTYNVAI EGASLVTVYR NQFGEIIKAK GVDGLIAELK AKNGGK* Computer analysis of this amino acid sequence gave the following results: Homology with a predicted ORF from N.meninsitidis (strain Al ORF91 shows 92.4% identity over a 92aa overlap with an ORF (ORF91a) from strain A of N. meningitidis: 10 20 30 40 50 60 orf91.pep MKKSSLISALGIGILSIGMAFAAPADAVSQIRQNATQVLSILKNGDANTARQKAEAYAIP I I II I : I I I I I II I I I I II I II II I I I I : I II I I II I I I I I I I : I I I I I I I I I II I I II I orf91a MKKSSFISALGIGILSIGMAFAAPADAVNQIRQNATQVLSILKSGDANTARQKAEAYAIP 10 20 30 40 50 60 70 80 90 orf91.pep YFDFQRMTALAVGNPWXTXSDXQKQALAXEFQP II I I I I I II I II I II I I II II I II I III orf91a YFDFQRMTALAVGNPWRTASDAQKQALAKEFQTLLIRTYSGTMLKLKNANVNVKDNPIVN 70 80 90 100 110 120 orf91a KGGKEIIVRAEVGVPGQKPVNMDFTTYQSGGKYRTYNVAIEGASLVTVYRNQFGEIIKAK 130 140 150 160 170 180 The complete length ORF9 la nucleotide sequence <SEQ ID 349> is: 1 ATGAAAAAAT CCTCCTTCAT CAGCGCATTG GGCATCGGTA TTTTGAGCAT 51 CGGCATGGCA TTTGCCGCCC CTGCCGACGC GGTAAACCAA ATCCGTCAAA 101 ACGCCACTCA AGTATTGAGC ATCTTAAAAA GCGGTGATGC CAACACCGCC 151 CGCCAAAAAG CCGAAGCCTA TGCGATTCCC TATTTCGATT TCCAACGTAT 201 GACCGCATTG GCGGTCGGCA ACCCTTGGCG CACCGCGTCC GACGCGCAAA 251 AACAAGCGTT GGCCAAAGAA TTTCAAACCC TGCTGATCCG CACCTATTCC 301 GGCACGATGC TGAAATTAAA AAACGCCAAC GTCAACGTCA AAGACAATCC 351 CATCGTCAAT AAAGGCGGCA AAGAAATCAT CGTCCGCGCC GAAGTCGGCG 401 TACCCGGGCA AAAACCCGTC AACATGGACT TCACCACCTA CCAAAGCGGC 451 GGTAAATACC GTACCTACAA CGTCGCCATC GAAGGCGCGA GCCTGGTTAC 501 CGTGTACCGC AACCAATTCG GCGAAATTAT CAAAGCGAAA GGCGTGGACG 551 GACTGATTGC CGAGTTGAAG GCTAAAAACG GCAGCAAGTA A This encodes a protein having amino acid sequence <SEQ ID 350>: 1 MKKSSFISAL GIGILSIGMA FAAPADAVNQ IRQNATQVLS ILKSGDANTA 51 RQKAEAYAIP YFDFQRMTAL AVGNPWRTAS DAQKQALAKE FQTLLIRTYS 101 GTMLKLKNAN VNVKDNPIVN KGGKEIIVRA EVGVPGQKPV NMDFTTYQSG 151 GKYRTYNVAI EGASLVTVYR NQFGEIIKAK GVDGLIAELK AKNGSK* ORF91a and ORF91-1 show 98.0% identity in 196 aa overlap: 10 20 30 40 50 60 orf91a. pep MKKSSFISALGIGILSIGMAFAAPADAVNQIRQNATQVLSILKSGDANTARQKAEAYAIP I II I I : I I II II II I I I I I I I II I II I I : II I II I II I I I I I I : I I I I I I I II I II II I I orf91-1 MKKSSLISALGIGILSIGMAFAAPADAVSQIRQNATQVLSILKNGDANTARQKAEAYAIP 10 20 30 40 50 60 70 80 90 100 110 120 orf91a . pep YFDFQRMTALAVGNPWRTASDAQKQALAKEFQTLLIRTYSGTMLKLKNANVNVKDNPIVN I I II I I I I I II II I I II I I II I II I I I I I I II I I II II I II I I I I I II I II I I II I I I I I orf91-1 YFDFQRMTALAVGNPWRTASDAQKQALAKEFQTLLIRTYSGTMLKLKNANVNVKDNPIVN 70 80 90 100 110 120 130 140 150 160 170 180 orf91a . pep KGGKEIIVRAEVGVPGQKPVNMDFTTYQSGGKYRTYNVAIEGASLVTVYRNQFGEIIKAK orf91-1 KGGKEIIVRAEVGVPGQKPVNMDFTTYQSGGKYRTYNVAIEGASLVTVYRNQFGEIIKAK 130 140 150 160 170 180 190 o f91a. pep GVDGLIAELKAKNGSKX II I II II I I I I I I I: I I orf91-l GVDGLIAELKAKNGGKX 190 Homology with a predicted ORF from N.gonorrhoeae ORF91 shows 84.8% identity over a 92aa overlap with a predicted ORF (ORF91.ng) from N gonorrhoeae: orf91. pep MKKSSLISALGIGILSIGMAFAAPADAVSQIRQNATQVLSILKNGDANTARQKAEAYAIP 60 Ml I MM I I I I I II II II I MMM I M I II I I I I MM MM I Ml I II I I IM orfθlng VKKSSFISALGIGILSIGMAFASPADAVGQIRQNATQVLTILKSGDAASARPKAEAYAVP 60 orf91. pep YFDFQRMTALAVGNPWXTXSDXQKQALAXEFQP 93 II I I II II II I I I II I I I I I II I II II I orf91ng YFDFQRMTALAVGNPWRTASDAQKQALAKEFQTLLIRTYSGTMLKFKNATVNVKDNPIVN 120 The complete length ORF91ng nucleotide sequence <SEQ LD 351> is predicted to encode a protein having amino acid sequence <SEQ ID 352>: 1 VKKSSFISAL GIGILSIGMA FASPADAVGQ IRQNATQVLT ILKSGDAASA 51 RPKAEAYAVP YFDFQRMTAL AVGNPWRTAS DAQKQALAKE FQTLLIRTYS 101 GTMLKFKNAT VNVKDNPIVN KGGKEIWRA EVGIPGQKPV NMDFTTYQSG 151 GKYRTYNVAI EGTSLVTVYR NQFGEIIKAK GIDGLIAELK AKNGGK* Further work revealed the complete nucleotide sequence <SEQ ID 353>: 1 ATGAAAAAAT CCTCCTTCAT CAGCGCATTG GGCATCGGTA TTTTGAGCAT 51 CGGCATGGCA TTTGCCTCCC CGGCCGACGC AGTGGGACAA ATCCGCCAAA 101 ACGCCACACA GGTTTTGACC ATCCTCAAAA GCGGCGACGC GGCTTCTGCA 151 CGCCCAAAAG CCGAAGCCTA TGCGGTTCCC TATTTCGATT TCCAACGTAT 201 GACCGCATTG GCGGTCGGCA ACCCTTGGCG TACCGCGTCC GACGCGCAAA 251 AACAAGCGTT GGCCAAAGAA TTTCAAACCC TGCTGATCCG CACCTATTCC 301 GGCACGATGC TGAAATTCAA AAACGCGACC GTCAACGTCA AAGACAATCC 351 CATCGTCAAT AAGGGCGGCA AGGAAATCGT CGTCCGTGCC GAAGTCGGCA 401 TCCCCGGTCA GAAGCCCGTC AATATGGACT TTACCACCTA CCAAAGCGGC 451 GGCAAATACC GTACCTACAA CGTCGCCATC GAAGGCACGA GCCTGGTTAC 501 CGTGTACCGC AACCAATTCG GCGAAATCAT CAAAGCCAAA GGCATCGACG 551 GGCTGATTGC CGAGTTGAAA GCCAAAAACG GCGGCAAATA A This corresponds to the amino acid sequence <SEQ ID 354; ORF91ng-l>: 1 MKKSSFISAL GIGILSIGMA FASPADAVGQ IRQNATQVLT ILKSGDAASA 51 RPKAEAYAVP YFDFQRMTAL AVGNPWRTAS DAQKQALAKE FQTLLIRTYS 101 GTMLKFKNAT VNVKDNPIVN KGGKEIWRA EVGIPGQKPV NMDFTTYQSG 151 GKYRTYNVAI EGTSLVTVYR NQFGEIIKAK GIDGLIAELK AKNGGK* ORF91ng-l and ORF91-1 show 92.3% identity in 196 aa overlap: 10 20 30 40 50 60 orf91-1.pep MKKSSLISALGIGILSIGMAFAAPADAVSQIRQNATQVLSILKNGDANTARQKAEAYAIP I I II I: I I II II I I I II II II I: I I I I I: I II II I II II : I I I: I I I l II I I I I: I orf91ng-l MKKSSFISALGIGILSIGMAFASPADAVGQIRQNATQVLTILKSGDAASARPKAEAYAVP 10 20 30 40 50 60 70 80 90 100 110 120 orf91-1. pep YFDFQRMTALAVGNPWRTASDAQKQALAKEFQTLLIRTYSGTMLKLKNANVNVKDNPIVN II I II I II I II I I I II I I I I I I I I I I I II I I I I I II I I I I I I I IM I I I: I II I I II II I orf91ng-l YFDFQRMTALAVGNPWRTASDAQKQALAKEFQTLLIRTYSGTMLKFKNATVNVKDNPIVN 70 80 90 100 110 120 130 140 150 160 170 180 orf91-l.pep KGGKEIIVRAEVGVPGQKPVNMDFTTYQΞGGKYRTYNVAIEGASLVTVYRNQFGEIIKAK I I I I I I M II II I : I I I I I I II II I I I I I I I II I I I II II II : II II I I I II I I I I M M orf91ng-l KGGKEIVVRAEVGIPGQKPVNMDFTTYQSGGKYRTYNVAIEGTSLVTVYRNQFGEIIKAK 130 140 150 160 170 180 190 orf91-1. pep GVDGLIAELKAKNGGKX I: I I I I II I I I I I I I I I orf91ng-l GIDGLIAELKAKNGGKX 190 In addition, ORF91ng-l shows homology to a hypothetical E.coli protein: sp|P45390|YRBC_ECOLI HYPOTHETICAL 24.0 KD PROTEIN IN MURA-RPON INTERGENIC REGION PRECURSOR (F211) >gi I 606130 (U18997) ORF_f211 [Escherichia coli] >gi 11789583 (AE000399) hypothetical 24.0 kD protein in murZ-rpoN intergenic region [Escherichia coli] Length = 211 Score = 70.6 bits (170), Expect = 6e-12 Identities = 42/137 (30%), Positives = 76/137 (54%), Gaps = 6/137 (4%) Query: 59 VPYFDFQRMTALAVGNPWRTASDAQKQALAKEFQTLLIRTYSGTMLKFKNATVNVKDNPI 118 +PY + AL +G +++A+ AQ++A F+ L + Y + + T + P Sbjct: 65 LPYVQVKYAGALVLGQYYKSATPAQREAYFAAFREYLKQAYGQALAMYHGQTYQIA—PE 122 Query: 119 VNKGGKEIV-VRAEVGIP-GQKPVNMDFTTYQSG—GKYRTYNVAIEGTSLVTVYRNQFG 174 G K IV +R + P G+ PV +DF ++ G ++ Y++ EG S++T +N++G Sbjct: 123 QPLGDKTIVPIRVTIIDPNGRPPVRLDFQWRKNSQTGNWQAYDMIAEGVSMITTKQNEWG 182 Query: 175 EIIKAKGIDGLIAELKA 191 +++ KGIDGL A+LK+ Sbjct: 183 TLLRTKGIDGLTAQLKS 199 Based on this analysis, including the presence of a putative leader sequence in the gonococcal protein, it is predicted that the proteins from N. meningitidis .and N.gonorrhoeae, .and their epitopes, could be useful antigens for vaccines or diagnostics, or for raising antibodies. Example 42 The following DNA sequence was identified in N. meningitidis <SEQ ID 355>: 1 ATGAAACACA TACTCCCCCT GATTGCCGCA TCCGCACTCT GCATTTCAAC 51 CGCTTCGGCA CATCCTGCCA GCGAACCGTC CACTCAAAAC GAAACCGCTA 101 TGATCACGCA TACCCTCATC TCAAAATACA GTTTTGGnnn nnnnnnnnnn 151 nnnnnnnnnn nnGCCATAAA AAGCAAAGGG ATGGACATTT TTGCCGTCAT 201 CGACCATCAG GAAGCCGCAC GCCGAAACGG CTTAACGATG CAGCCGGCAA 251 AAGTCATCGT CTTCGGCACG CCCAAAGCCG GCACGCCGCT GATGGTCAAA 301 GACCCCGCCT TCGCCCTGCA ACTGCCCCTA CGCGTCCTCG TTACCGAAAC 351 GGACGGCAAA GTACGCGCCG CCTATACCGA TACGCGCGCC CTCATCGCCG 401 GCAGCCGCAT CGGTTTCGAC GAAGTGGCAA ACACTTTGGC AAACGCCGAA 451 AAACTGATAC AAAAAACCGT AGGCGAATAA This corresponds to the .amino acid sequence <SEQ ID 356; ORF97>: 1 MKHILPLIAA SALCISTASA HPASEPSTQN ETAMITHTLI SKYSFGXXXX 51 XXXXAIKSKG MDIFAVIDHQ EAARRNGLTM QPAKVIVFGT PKAGTPLMVK 101 DPAFALQLPL RVLVTETDGK VRAAYTDTRA LIAGSRIGFD EVANTLANAE 151 KLIQKTVGE* Further work revealed the complete nucleotide sequence <SEQ ID 357>: 1 ATGAAACACA TACTCCCCCT GATTGCCGCA TCCGCACTCT GCATTTCAAC 51 CGCTTCGGCA CATCCTGCCA GCGAACCGTC CACCCAAAAC GAAACCGCTA 101 TGACCACGCA TACCCTCACC TCAAAATACA GTTTTGACGA AACCGTCAGC 151 CGCCTTGAAA CCGCCATAAA AAGCAAAGGG ATGGACATTT TTGCCGTCAT 201 CGACCATCAG GAAGCCGCCC GCCGAAACGG CTTAACGATG CAGCCGGCAA 251 AAGTCATCGT CTTCGGCACG CCCAAAGCCG GCACGCCGCT GATGGTCAAA 301 GACCCCGCCT TCGCCCTGCA ACTGCCCCTA CGCGTCCTCG TTACCGAAAC 351 GGACGGCAAA GTACGCGCCG CCTATACCGA TACGCGCGCC CTCATCGCCG 401 GCAGCCGCAT CGGTTTCGAC GAAGTGGCAA ACACTTTGGC AAACGCCGAA 451 AAACTGATAC AAAAAACCGT AGGCGAATAA This corresponds to the amino acid sequence <SEQ ED 358; ORF97-l>: 1 MKHILPLIAA SALCISTASA HPASEPSTQN ETAMTTHTLT SKYSFDETVS 51 RLETAIKSKG MDIFAVIDHQ EAARRNGLTM QPAKVIVFGT PKAGTPLMVK 101 DPAFALQLPL RVLVTETDGK VRAAYTDTRA LIAGSRIGFD EVANTLANAE 151 KLIQKTVGE* Computer analysis of this amino acid sequence gave the following results: Homology with a predicted ORF from N.meninsitidis (strain Al ORF97 shows 88.7% identity over a 159aa overlap with an ORF (ORF97a) from strain A of N. meningitidis: 10 20 30 40 50 60 orf97.pep MKHILPLIAASALCISTASAHPASEPSTQNETAMITHTLISKYSFGXXXXXXXXAIKSKG I I I I I I I I I I I II I II I II I I I: I I I I I I I I I I I I II II : : I II I I I orf97a MXHILPLXXASALCISTASXHPASEPQTQNETAMTTHTLTSKYSFDETVSRLETAIKSKG 10 20 30 40 50 60 70 80 90 100 110 120 orf97. pep MDIFAVIDHQEAARRNGLTMQPAKVIVFGTPKAGTPLMVKDPAFALQLPLRVLVTETDGK II I I I I II II I I I I I I I II II II II I II I I I II I II I I I II I I I I I I I I I I I I I I I I I I orf97a MDIFAVIDHQEAARRNGLTMQPAKVIVFGTPKAGTPLMVKDPAFALQLPLRVXVTETDGK 70 80 90 100 110 120 130 140 150 160 orf97. pep VRAAYTDTRALIAGSRIGFDEVANTLANAEKLIQKTVGEX I I I II II I I I I I I I I I I II I II I II II I II I II I II : I I I orf97a VRAAYTDTRALIAGSRIGFDEVANTLANAEKLIQKTIGEX 130 140 150 160 The complete length ORF97a nucleotide sequence <SEQ ID 359> is: 1 ATGANACACA TACTCCCCCT GANTGNCGCA TCCGCACTCT GCATTTCAAC 51 CGCTTCGGNN CATCCTGCCA GCGAACCGCA AACCCAAAAC GAAACCGCTA 101 TGACCACGCA TACCCTCACC TCAAAATACA GTTTTGACGA AACCGTCAGC 151 CGCCTTGAAA CCGCCATAAA AAGCAAAGGG ATGGACATTT TTGCCGTCAT 201 CGACCATCAG GAAGCCGCCC GCCGAAACGG CTTAACGATG CAGCCGGCAA 251 AAGTCATCGT CTTCGGCACG CCCAAAGCCG GTACGCCGCT GATGGTCAAA 301 GACCCCGCCT TCGCCCTGCA ACTGCCCCTG CGCGTCNTCG TTACCGAAAC 351 GGACGGCAAA GTACGCGCCG CCTATACCGA TACGCGCGCC CTCATCGCCG 401 GCAGCCGCAT CGGTTTCGAC GAAGTGGCAA ACACTTTGGC AAACGCCGAA 451 AAACTGATAC AAAAAACCAT AGGCGAATAA This encodes a protein having amino acid sequence <SEQ ID 360>: 1 MXHILPLXXA SALCISTASX HPASEPQTQN ETAMTTHTLT SKYSFDETVS 51 RLETAIKSKG MDIFAVIDHQ EAARRNGLTM QPAKVIVFGT PKAGTPLMVK 101 DPAFALQLPL RVXVTETDGK VRAAYTDTRA LIAGSRIGFD EVANTLANAE 151 KLIQKTIGE* ORF97a and ORF97-1 show 95.6% identity in 159 aa overlap: 10 20 30 40 50 60 orf97a. pep MXHILPLXXASALCISTASXHPASEPQTQNETAMTTHTLTSKYSFDETVSRLETAIKSKG I II I II I I I II I I II I II I I I I: I II I I II II II I I I I II II I I I I II I I I I I I II orf97-1 MKHILPLIAASALCISTASAHPASEPSTQNETAMTTHTLTSKYSFDETVSRLETAIKSKG 10 20 30 40 50 60 70 80 90 100 110 120 orf97a . pep MDIFAVIDHQEAARRNGLTMQPAKVIVFGTPKAGTPLMVKDPAFALQLPLRVXVTETDGK II II I I I II II I I I II I I I I I I I I I II I I I I I II I I II II II II I I I I I I I I I I I I I I I orf97-1 MDIFAVIDHQEAARRNGLTMQPAKVIVFGTPKAGTPLMVKDPAFALQLPLRVLVTETDGK 70 80 90 100 110 120 130 140 150 160 orf97a. pep VRAAYTDTRALIAGSRIGFDEVANTLANAEKLIQKTIGEX I I II II II I I II I II II II II I I I II I II II II I II: I I I or 97-1 VRAAYTDTRALIAGSRIGFDEVANTLANAEKLIQKTVGEX 130 140 150 160 Homology with a predicted ORF from N.sonorrhoeae ORF97 shows 88.1% identity over a 159aa overlap with a predicted ORF (ORF97.ng) from N. gonorrhoeae: orf97.pep MKHILPLIAASALCISTASAHPASEPSTQNETAMITHTLISKYSFGXXXXXXXXAIKSKG 60 I I I I I I I I II I : I I I I I I I II I :: I I I II II I I I I I I I I II : M I II II orf97ng MKHILPPIAASAFCISTASAHPAGKPPTQNETAMTTHTLTSKYSFDETVSRLETAIKSKG 60 orf97.pep MDIFAVIDHQEAARRNGLTMQPAKVIVFGTPKAGTPLMVKDPAFALQLPLRVLVTETDGK 120 I I I I I I I II II I II I I I I I II I I I I II I I I I I II I II II I I I I I II II I I I I I II I II I I orf97ng MDIFAVIDHQEAARRNGLTMQPAKVIVFGTPKAGTPLMVKDPAFALQLPLRVLVTETDGK 120 orf97.pep VRAAYTDTRALIAGSRIGFDEVANTLANAEKLIQKTVGE 159 I IM I II II I I M IMM II I II I II II I I I I I I I I I orf97ng VRTAYTDTRALIVGSRISFDEVANTLANAEKLIQKTVGE 159 The complete length ORF97ng nucleotide sequence <SEQ ID 361> is predicted to encode a protein having amino acid sequence <SEQ ID 362>: 1 MKHILPPIAA SAFCISTASA HPAGKPPTQN ETAMTTHTLT SKYSFDETVS 51 RLETAIKSKG MDIFAVIDHQ EAARRNGLTM QPAKVIVFGT PKAGTPLMVK 101 DPAFALQLPL RVLVTETDGK VRTAYTDTRA LIVGSRISFD EVANTLANAE 151 KLIQKTVGE* Further work revealed the complete nucleotide sequence <SEQ LD 363>: 1 ATGAAACACA TACTCCCcct gatcgccgca TccgcactCT GCATTTCAAC 51 CGCTTCGGCA CACCCTGCCG GCAAACCGCC CACCCAAAAC GAAACCGCTA 101 TGACCACGCA CACCCTCACC TCGAAATACA GTTTTGACGA AACCGTCAGC 151 CGCCTTGAAA CCGCCATAAA AAGCAAAGGG ATGGACATTT TTGCCGTCAT 201 CGACCATCAG GAAGCGGCAC GCCGAAACGG CCTGACCATG CAGCCGGCAA 251 AAGTCATCGT CTTCGGCACG CCCAAGGCCG GTACGCCgct GATGGTCAAA 301 GACCCCGCCT TCGCCCTGCA ACTGCCCCTG CGCGTCCTCG TTACCGAAAC 351 GGACGGCAAA GTACGCACCG CCTATACCGA TACGCGCGCC CTCATCGTCG 401 GCAGCCGCAT CAGTTTCGAC GAAGTGGCAA ACACTTTGGC AAACGCCGAA 451 AAACTGATAC AAAAAACCGT AGGCGAATAA This corresponds to the .amino acid sequence <SEQ ID 364; ORF97ng-l>: 1 MKHILPLIAA SALCISTASA HPAGKPPTQN ETAMTTHTLT SKYSFDETVS 51 RLETAIKSKG MDIFAVIDHQ EAARRNGLTM QPAKVIVFGT PKAGTPLMVK 101 DPAFALQLPL RVLVTETDGK VRTAYTDTRA LIVGSRISFD EVANTLANAE 151 KLIQKTVGE* ORF97ng-l and ORF97-1 show 96.2% identity in 159 aa overlap: 10 20 30 40 50 60 orf97-1. pep MKHILPLIAASALCISTASAHPASEPSTQNETAMTTHTLTSKYSFDETVSRLETAIKSKG I I II I I I I I I I II II I I I II I I I :: I I II II II II II II I II I II I I I I I I I II I I II I orf97ng-l MKHILPLIAASALCISTASAHPAGKPPTQNETAMTTHTLTSKYSFDETVSRLETAIKSKG 10 20 30 40 50 60 70 80 90 100 110 120 orf97-1. pep MDIFAVIDHQEAARRNGLTMQPAKVIVFGTPKAGTPLMVKDPAFALQLPLRVLVTETDGK II I II I II I II I I I I II II I II I I II I I I I I I II I II II II I I I I I I II I I I II I II II I orf97ng-l MDIFAVIDHQEAARRNGLTMQPAKVIVFGTPKAGTPLMVKDPAFALQLPLRVLVTETDGK 70 80 90 100 110 120 130 140 150 160 orf97-1. pep VRAAYTDTRALIAGSRIGFDEVANTLANAEKLIQKTVGEX II: I I I I I II I I: II II: I II I I I I I I I II I II I I I I I I I orf97ng-l VRTAYTDTRALIVGSRISFDEVANTLANAEKLIQKTVGEX 130 140 150 160 Based on this analysis, including the presence of a putative leader sequence in the gonococcal protein, it was predicted that the proteins from N. meningitidis and N.gonorrhoeae, and their epitopes, could be useful antigens for vaccines or diagnostics, or for raising antibodies. ORF97-1 (15.3kDa) was cloned in pET and pGex vectors and expressed in E.coli, as described above. The products of protein expression and purification were analyzed by SDS-PAGE. Figures 12A & 12B show, repsectively, the results of affinity purification of the GST-fiision and His-fusion proteins. Purified GST-fusion protein was used to immunise mice, whose sera were used for Western Blot (Figure 12C), ELISA (positive result), and FACS analysis (Figure 12D). These experiments confirm that ORF97-1 is a surface-exposed protein, and that it is a useful immunogen. Figure 12E shows plots of hydrophilicity, antigenic index, and AMPHI regions for ORF97-1. Example 43 The following DNA, believed to be complete, sequence was identified in N. meningitidis <SEQ ID 365>: 1 ATGGCTTTTA TTACGCGCTT ATTCAAAAGC AGTAAATGGC TGATTGTGCC 51 GCTGATGCTC CCCGCCTTTC AGAATGTGGC GGCGGAGGGG ATAGATGTGA 101 GCCGTGCCGA AGCGAGGATA ACCGACGGCG GGCAGCTTTC CATCAGCAGC 151 CGCTTCCAAA CCGAGCTGCC CGACCAGCTC CAACAGGCGT TGCGCCGGGg 201 CGTGCCGCTC AACTTTACCT TAAGCTGGCA GCTTTCCGCC CCGATAATCG 251 CTTCTTATCG GTTTAAATTG GGGCAACTGA TTGGCGATGA CGACaATATT 301 GACTACAAAC TGAGTTTCCA TCCGCTGACc AaACGCTACC GCGTTACCgT 351 CGgCGCGTTT TCGACAGACT ACGACACCTT GGATGCGGCA TTGCGCGCGA 401 CCGGCGCGGT TGCCAACTGG AAAGTCCTGA ACAAAGGCGC GCTGTCCGGT 451 GCGGAAGCAG GGGAAACCAA GGCGGAAATC CGCCTGACGC TGTCCACTTC 501 AAAACTGCCC AAGCCTTTTC AAATCAATGC ATTGACTTCT CAAAACTGGC 551 ATTTGGATTC GGGTTGGAAA CCTCTAAACA TCATCGGGAA CAAATAA This corresponds to the amino acid sequence <SEQ LD 366; ORF106>: 1 MAFITRLFKS SKWLIVPLML PAFQNVAAEG IDVSRAEARI TDGGQLSISS 51 RFQTELPDQL QQALRRGVPL NFTLSWQLSA PIIASYRFKL GQLIGDDDNI 101 DYKLSFHPLT KRYRVTVGAF STDYDTLDAA LRATGAVANW KVLNKGALSG 151 AEAGETKAEI RLTLSTSKLP KPFQINALTS QNWHLDSGWK PLNIIGNK* Further work revealed the following DNA sequence <SEQ LD 367>: 1 ATGGCTTTTA TTACGCGCTT ATTCAAAAGC AGTAAATGGC TGATTGTGCC 51 GCTGATGCTC CCCGCCTTTC AGAATGTGGC GGCGGAGGGG ATAGATGTGA 101 GCCGTGCCGA AGCGAGGATA ACCGACGGCG GGCAGCTTTC CATCAGCAGC 151 CGCTTCCAAA CCGAGCTGCC CGACCAGCTC CAACAGGCGT TGCGCCGGGG 201 CGTGCCGCTC AACTTTACCT TAAGCTGGCA GCTTTCCGCC CCGATAATCG 251 CTTCTTATCG GTTTAAATTG GGGCAACTGA TTGGCGATGA CGACAATATT 301 GACTACAAAC TGAGTTTCCA TCCGCTGACC AACCGCTACC GCGTTACCGT 351 CGGCGCGTTT TCGACAGACT ACGACACCTT GGATGCGGCA TTGCGCGCGA 401 CCGGCGCGGT TGCCAACTGG AAAGTCCTGA ACAAAGGCGC GCTGTCCGGT 451 GCGGAAGCAG GGGAAACCAA GGCGGAAATC CGCCTGACGC TGTCCACTTC 501 AAAACTGCCC AAGCCTTTTC AAATCAATGC ATTGACTTCT CAAAACTGGC 551 ATTTGGATTC GGGTTGGAAA CCTCTAAACA TCATCGGGAA CAAATAA This corresponds to the amino acid sequence <SEQ LD 368; ORF106-1>: 1 MAFITRLFKS SKWLIVPLML PAFQNVAAEG IDVSRAEARI TDGGQLSISS 51 RFQTELPDQL QQALRRGVPL NFTLSWQLSA PIIASYRFKL GQLIGDDDNI 101 DYKLSFHPLT NRYRVTVGAF STDYDTLDAA LRATGAVANW KVLNKGALSG 151 AEAGETKAEI RLTLSTSKLP KPFQINALTS QNWHLDSGWK PLNIIGNK* Computer analysis of this amino acid sequence gave the following results: Homology with a predicted ORF from N.meninsitidis (strain A) ORF106 shows 87.4% identity over a 199aa overlap with an ORF (ORF106a) from strain A of N. meningitidis: 10 20 30 40 50 59 orf106. pep MAFITRLFKSSK-WLIVPLMLPAFQNVAAEGIDVSRAEARITDGGQLSISSRFQTELPDQ II I I II I II I I I I :: II : : :: I I II I II I II I I I I : I I II I I I II I I I I I I I orf106a MAFITRLFKSIKQWLVLLPMLSVLPDAAAEGIDVSRAEARIXDGGQLSXXSRFQTELPDQ 10 20 30 40 50 60 60 70 80 90 100 110 119 orf106. pep LQQALRRGVPLNFTLSWQLSAPIIASYRFKLGQLIGDDDNIDYKLSFHPLTKRYRVTVGA II I III II II I I II I I II I I I I I I I II I I I I I II II I I II I I I : II I I I I I I orfl06a LQXAXXRGVXLNXTLXWQLSAPIIASYRFXLGQLIGDDDXIDYKLSFHPLTNRYRVTVGA 70 80 90 100 110 120 120 130 140 150 160 170 179 orf106. pep FSTDYDTLDAALRATGAVANWKVLNKGALSGAEAGETKAEIRLTLSTSKLPKPFQINALT III II I I I I I I I I I II II I I I II I II II I I I I I I I I I I I I I I II II I I I I II I I II II I orf106a FSTXYDTLDAALRATGAVANWKVLNKGALSGAEAGETKAEIRLTLSTSKLPKPFQINALT 130 140 150 160 170 180 180 190 199 orfl06.pep SQNWHLDSGWKPLNIIGNKX II I II II II I I I I II II I I I orfl06a SQNWHLDSGWKPLNIIGNKX 190 200 Due to the K»N substitution at residue 111 , the homology between ORF 106a and ORF 106- 1 is 87.9% over the same 199 aa overlap. The complete length ORF 106a nucleotide sequence <SEQ LD 369> is: 1 ATGGCTTTTA TTACGCGCTT ATTCAAAAGC ATTAAACAAT GGCTTGTGCT 51 GCTGCCGATG CTTTCCGTTT TGCCGGACGC GGCGGCGGAG GGGATAGATG 101 TGAGCCGCGC CGAAGCGAGG ATAANCGACG GCGGGCAGCT TTCCATNAGN 151 AGCCGCTTCC AAACCGAGCT GCCCGACCAG CTCCAANNNG CGNNGNGCCG 201 GGGCGTGNCG CTCAACTNTA CCTTAAGNTG GCAGCTTTCC GCCCCGATAA 251 TCGCTTCTTA TCGGTTTNAA TTGGGGCAAC TGATTGGCGA TGACGACNAT 301 ATTGACTACA AACTGAGTTT CCATCCGCTG ACCAACCGCT ACCGCGTTAC 351 CGTCGGCGCG TTTTCGACAG ANTACGACAC CTTGGATGCG GCATTGCGCG 401 CGACCGGCGC GGTTGCCAAC TGGAAAGTCC TGAACAAAGG CGCGCTGTCC 451 GGTGCGGAAG CAGGGGAAAC CAAGGCGGAA ATCCGCCTGA CGCTGTCCAC 501 TTCAAAACTG CCCAAGCCTT TTCAAATCAA TGCATTGACT TCTCAAAACT 551 GGCATTTGGA TTCGGGTTGG AAACCTCTAA ACATCATCGG GAACAAATAA This encodes a protein having amino acid sequence <SEQ ID 370>: 1 MAFITRLFKS IKQWLVLLPM LSVLPDAAAE GIDVSRAEAR IXDGGQLSXX 51 SRFQTELPDQ LQXAXXRGVX LNXTLXWQLS APIIASYRFX LGQLIGDDDX 101 IDYKLSFHPL TNRYRVTVGA FSTXYDTLDA ALRATGAVAN WKVLNKGALS 151 GAEAGETKAE IRLTLSTSKL PKPFQINALT SQNWHLDSGW KPLNIIGNK* Homology with a predicted ORF from N.sonorrhoeae ORF 106 shows 90.5% identity over a 199aa overlap with a predicted ORF (ORFlOό.ng) from N. gonorrhoeae: orf106. pep MAFITRLFKSSK-WLIVPLMLPAFQNVAAEGIDVSRAEARITDGGQLSISΞRFQTELPDQ 59 I I II I I II I I I I I : : : I :: :: II II I :: I II I I I I I I I : I I I I I I I I I I I I II orfl06ng MAFITRLFKSIKQWLVLLPILSVLPDAAAEGIAATRAEARITDGGRLSISSRFQTELPDQ 60 orf106. pep LQQALRRGVPLNFTLSWQLSAPIIASYRFKLGQLIGDDDNIDYKLSFHPLTKRYRVTVGA 119 I I I I I I I II I I I I I I I I II I I I II I II II I I I II I I I I II I I I I I II I I I : I I I I I II I orfl06ng LQQALRRGVPLNFTLSWQLSAPTIASYRFKLGQLIGDDDNIDYKLSFHPLTNRYRVTVGA 120 orf106. pep FSTDYDTLDAALRATGAVANWKVLNKGALSGAEAGETKAEIRLTLSTSKLPKPFQINALT 179 I I I II I II II I I I II I II I I I II I I I I I I I I I II I I I II I I I II I I I I I I I I I II I I I I I orfl06ng FSTDYDTLDAALRATGAVANWKVLNKGALSGAEAGETKAEIRLTLSTSKLPKPFQINALT 180 orfl06.pep SQNWHLDSGWKPLNIIGNK 198 III I II II I II I I I I I I II orflOδng SQNWHLDSGWKPLNIIGNK 199 Due to the K»N substitution at residue 111, the homology between ORF106ng and ORF 106-1 is 91.0% over the same 199 aa overlap. The complete length ORFlOόng nucleotide sequence <SEQ ID 371> is: 1 ATGGCTTTTA TTACGCGCTT ATTCAAAAGC ATTAAACAAT GGCTTGTGCT 51 GTTGCCGATA CTCTCCGTTT TGCCGGACGC GGCGGCGGAG GGCATTGCCG 101 CGACCCGCGC CGAAGCGAGG ATAACCGACG GCGGGCGGCT TTCCATCAGC 151 AGCCGCTTCC AAACCGAGCT GCCCGACCAG CTCCAACAGG CGTTGCGCCG 201 GGGCGTACCG CTCAACTTTA CCTTAAGCTG GCAGCTTTCC GCCCCGACAA 251 TCGCTTCTTA TCGGTTTAAA TTGGGGCAAC TGATTGGCGA TGACGACAAT 301 ATTGACTACA AACTAAGTTT CCATCCGCTG ACCAACCGCT ACCGCGTTAC 351 CGTCGGCGCA TTTTCCACCG ATTACGACAC TTTGGATGCG GCATTGCGCG 401 CGACCGGCGC GGTTGCCAAC TGGAAAGTCC TGAACAAAGG CGCGTTGTCC 451 GGTGCGGAAG CAGGGGAAAC CAAGGCGGAA ATCCGCCTGA CGCTGTCCAC 501 TTCAAAACTG CCCAAGCCTT TCCAAATCAA CGCATTGACT TCTCAAAACT 551 GGCATTTGGA TTCGGGTTGG AAACCTCTAA ACATCATCGG GAACAAATAA This encodes a protein having amino acid sequence <SEQ ID 372>: 1 MAFITRLFKS IKQWLVLLPI LSVLPDAAAE GIAATRAEAR ITDGGRLSIS 51 SRFQTELPDQ LQQALRRGVP LNFTLSWQLS APTIASYRFK LGQLIGDDDN 101 IDYKLSFHPL TNRYRVTVGA FSTDYDTLDA ALRATGAVAN WKVLNKGALS 151 GAEAGETKAE IRLTLSTSKL PKPFQINALT SQNWHLDSGW KPLNIIGNK* Based on this analysis, including the presence of a putative leader sequence in the gonococcal protein, it was predicted that the proteins from N. meningitidis and N.gonorrhoeae, and their epitopes, could be useful antigens for vaccines or diagnostics, or for raising antibodies. ORF 106-1 (18kDa) was cloned in pET and pGex vectors and expressed in E.coli, as described above. The products of protein expression and purification were analyzed by SDS-PAGE. Figure 13A shows the results of affinity purification of the His-fusion protein, and Figure 13B shows the results of expression of the GST-fusion in E.coli. Purified His-fusion protein was used to immunise mice, whose sera were used for FACS analysis (Figure 13C) These experiments confirm that ORF 106-1 is a surface-exposed protein, and that it is a useful immunogen. Example 44 The following DNA sequence, believed to be complete, w.as identified in N. meningitidis <SEQ LD 373>: 1 ATGGACACAA AAGAAATCCT CGG.TACGCG GcAGGcTCGA TCGGCAGCGC 51 GGTTTTAGCC GTCATCATCc TGCCGCTGCT GTCGTGGTAT TTCCCCGCCG 101 ACGACATCGG GCGCATCGTG CTGATGCAGA CGGCGGCGGG GCTgACGGTG 151 TCGGTGTTGT GCCTCGGGCT GGATCAGGCA TACGTCCGCG AATACTATGC 201 CACCGCCGAC AAAGACAcCT TGTTCAAAAC CCTGTTCCTG CCGCCGCTGC 251 TGTCTGCCGC CGCGATAGCC GCCCTGCTGC TTTCCCGCCC GTCCCTGCCG 301 TCTGAAATCC TGTTTTCACT CGACGATGCC gCCGCCGGCa TCGGGCTGGT 351 GCTGTTTGAA CtGAGCTTCC TGCCCATCCG cTTTCTCTTA CTGGTTTTGC 401 GTATGGAAGG ACGCGCCcTT GCCTTTTCGT CCGCGCAACT CGTGCcCAAG 451 CTCGCCATCC TGCTGCTG.T GCCGCTGACG GTCGGGCTGC TGCACTTTCC 501 AGCGAACACC GCCGTCCTGA CCGCCGTTTA CGCGCTGGCA AACCTTGCCG 551 CCGCCGCCTT TTTGCTGTTT CAAAACCGAT GCCGTCTGAA GGCCGTCCGG 601 CACGCACCGT TTTCGCCCGC CGTCCTGCAC CGGGGG.TGC GCTACGGCAT 651 ACCGATCGCA CTGAGCAGCA TCGCCTATTG GGGGCTGGCA TCCGCCGACC 701 GTTTGTTCCT GAAAAAATAT GCCGGCCTGG AACAGCTCGG CGTTTATTCG 751 ATGGGTATTT CGTTCGGCGG GGCGGCATTA TTGTTCCAAA GCATCTTTTC 801 AACGGTCTGG ACACCGTATA TTTTCCGCGC AATCGAAGAA AACGCCCCGC 851 CCGCTCGCCT CTCGGCAACG GCAGAATCCG CCGCCGCCCT GCTTGCCTCC 901 GCCCTCTGC. TGACCGGCAT TTTCTCGCCC CTTGCCTCCC TCCTGCTGCC 951 GGAAAACTAC GCCGCCGTCC GGTTTATCGT CGTATCGTGT ATG.TGCCGC 1001 CGCTGTTTTG CACGCTGGCG GAAATCAGCG GCATCGGTTT GAACGTCGTT 1051 CGCAAAACGC GCCCGATCGC GCTCGCCACC TTGGGCGCGC TGGCGGCAAA 1101 CCTGCTGCTG CTGGGGCTTG ACCGTGCCGT ACCGGCGAGG CCGCC.GGCG 1151 CGGCGGTTGC CTGTGCCGCC TCATTCTGGC TGTTTTTTGC CTTCAAGACC 1201 GAAAGCTCyT GCCGCCTGTG GCAGCCGCTC AAACGCCTGC CGCTTTATCT 1251 GCACACATTG TTCTGCCTGA CCTCCTCGGC GGCCTACACC TGCTTCGGCA 1301 CGCCGGCAAA CTATCCCCTG TTTGCCGGCG TATGGGCGGC ATATCTGGCA 1351 GGCTGCATCC TGCGCCACCG GAAAGATTTG CACAAACTGT TTCATTATTT 1401 GAAAAAACAA GGTTTCCCAT TATGA This corresponds to the amino acid sequence <SEQ ID 374; ORF10>: 1 MDTKEILXYA AGSIGSAVLA VIILPLLSWY FPADDIGRIV LMQTAAGLTV 51 SVLCLGLDQA YVREYYATAD KDTLFKTLFL PPLLSAAAIA ALLLSRPSLP 101 SEILFSLDDA AAGIGLVLFE LSFLPIRFLL LVLRMEGRAL AFSSAQLVPK 151 LAILLLXPLT VGLLHFPANT AVLTAVYALA NLAAAAFLLF QNRCRLKAVR 201 HAPFSPAVLH RGXRYGIPIA LSSIAYWGLA SADRLFLKKY AGLEQLGVYS 251 MGISFGGAAL LFQSIFSTVW TPYIFRAIEE NAPPARLSAT AESAAALLAS 301 ALCXTGIFSP LASLLLPENY AAVRFIWSC MXPPLFCTLA EISGIGLNW 351 RKTRPIALAT LGALAANLLL LGLDRAVPAR PXGAAVACAA SFWLFFAFKT 401 ESSCRLWQPL KRLPLYLHTL FCLTSSAAYT CFGTPANYPL FAGVWAAYLA 451 GCILRHRKDL HKLFHYLKKQ GFPL* Further sequence analysis revealed the complete DNA sequence<SEQ ID 375> to be: 1 ATGGACACAA AAGAAATCCT CGGCTACGCG GCAGGCTCGA TCGGCAGCGC 51 GGTTTTAGCC GTCATCATCC TGCCGCTGCT GTCGTGGTAT TTCCCCGCCG 101 ACGACATCGG GCGCATCGTG CTGATGCAGA CGGCGGCGGG GCTGACGGTG 151 TCGGTGTTGT GCCTCGGGCT GGATCAGGCA TACGTCCGCG AATACTATGC 201 CACCGCCGAC AAAGACACCT TGTTCAAAAC CCTGTTCCTG CCGCCGCTGC 251 TGTCTGCCGC CGCGATAGCC GCCCTGCTGC TTTCCCGCCC GTCCCTGCCG 301 TCTGAAATCC TGTTTTCACT CGACGATGCC GCCGCCGGCA TCGGGCTGGT 351 GCTGTTTGAA CTGAGCTTCC TGCCCATCCG CTTTCTCTTA CTGGTTTTGC 401 GTATGGAAGG ACGCGCCCTT GCCTTTTCGT CCGCGCAACT CGTGCCCAAG 451 CTCGCCATCC TGCTGCTGCT GCCGCTGACG GTCGGGCTGC TGCACTTTCC 501 AGCGAACACC GCCGTCCTGA CCGCCGTTTA CGCGCTGGCA AACCTTGCCG 551 CCGCCGCCTT TTTGCTGTTT CAAAACCGAT GCCGTCTGAA GGCCGTCCGG 601 CACGCACCGT TTTCGCCCGC CGTCCTGCAC CGGGGGCTGC GCTACGGCAT 651 ACCGATCGCA CTGAGCAGCA TCGCCTATTG GGGGCTGGCA TCCGCCGACC 701 GTTTGTTCCT GAAAAAATAT GCCGGCCTGG AACAGCTCGG CGTTTATTCG 751 ATGGGTATTT CGTTCGGCGG GGCGGCATTA TTGTTCCAAA GCATCTTTTC 801 AACGGTCTGG ACACCGTATA TTTTCCGCGC AATCGAAGAA AACGCCCCGC 851 CCGCCCGCCT CTCGGCAACG GCAGAATCCG CCGCCGCCCT GCTTGCCTCC 901 GCCCTCTGCC TGACCGGCAT TTTCTCGCCC CTTGCCTCCC TCCTGCTGCC 951 GGAAAACTAC GCCGCCGTCC GGTTTATCGT CGTATCGTGT ATGCTGCCGC 1001 CGCTGTTTTG CACGCTGGCG GAAATCAGCG GCATCGGTTT GAACGTCGTC 1051 CGCAAAACGC GCCCGATCGC GCTCGCCACC TTGGGCGCGC TGGCGGCAAA 1101 CCTGCTGCTG CTGGGGCTTG CCGTGCCGTC CGGCGGCGCG CGCGGCGCGG 1151 CGGTTGCCTG TGCCGCCTCA TTCTGGCTGT TTTTTGCCTT CAAGACCGAA 1201 AGCTCCTGCC GCCTGTGGCA GCCGCTCAAA CGCCTGCCGC TTTATCTGCA 1251 CACATTGTTC TGCCTGACCT CCTCGGCGGC CTACACCTGC TTCGGCACGC 1301 CGGCAAACTA TCCCCTGTTT GCCGGCGTAT GGGCGGCATA TCTGGCAGGC 1351 TGCATCCTGC GCCACCGGAA AGATTTGCAC AAACTGTTTC ATTATTTGAA 1401 AAAACAAGGT TTCCCATTAT GA This corresponds to the amino acid sequence <SEQ ID 376; ORF10-1>: 1 MDTKEILGYA AGSIGSAVLA VIILPLLSWY FPADDIGRIV LMQTAAGLTV 51 SVLCLGLDQA YVREYYATAD KDTLFKTLFL PPLLSAAAIA ALLLSRPSLP 101 SEILFSLDDA AAGIGLVLFE LSFLPIRFLL LVLRMEGRAL AFSSAQLVPK 151 LAILLLLPLT VGLLHFPANT AVLTAVYALA NLAAAAFLLF QNRCRLKAVR 201 HAPFSPAVLH RGLRYGIPIA LSSIAYWGLA SADRLFLKKY AGLEQLGVYS 251 MGISFGGAAL LFQSIFSTVW TPYIFRAIEE NAPPARLSAT AESAAALLAS 301 ALCLTGIFSP LASLLLPENY AAVRFIWSC MLPPLFCTLA EISGIGLNW 351 RKTRPIALAT LGALAANLLL LGLAVPSGGA RGAAVACAAS FWLFFAFKTE 401 SSCRLWQPLK RLPLYLHTLF CLTSSAAYTC FGTPANYPLF AGVWAAYLAG 451 CILRHRKDLH KLFHYLKKQG FPL* Computer analysis of this amino acid sequence gave the following results: Prediction ORF 10-1 is predicted to be the precursor of .an integral membrane protein, since it comprises several (12-13) potential transmembrane segments, and a probable cleavable signal peptide Homology with EpsM from Streptococcus thermophilus (accession number U40830). ORFIO shows homology with the epsM gene of S. thermophilus, which encodes a protein of a size similar to ORFIO and is involved in expolysaccharide synthesis. Other homologies are with prokaryotic membrane proteins: Identitxes = (25%) Query: 213 LRYGIPLALSSLAYWGLASADRLFLKKYAGLEQLGVYSMGISFGGAALLLQSIFSTVW 270 L Y +PL SS+ +W L ++ R F+ + G G+ ++ + +IF+ W Sbjct: 210 LYYALPLIPSSILWWLLNASSRYFVLFFLGAGANGLLAVATKIPSIISIFNTIFTQAW 267 Identities = 15/57 (26%), Positives = 31/57 (54%) Query: 7 LGYAAGSIGSAVLAVIILPLLSWYFPADDIGRIVLMQTAAGLTVSVLCLGLDQAYVR 63 L + G++GS +L +++PL ++ + G L QT A L + ++ + + A +R Sbjct: 12 LVFTIGNLGSKLLVFLLVPLYTYAMTPQEYGMADLYQTTANLLLPLITMNVFDATLR 68 Identities = 16/96 (16%), Positives = 36/96 (37%) Query: 307 IFSPLASLLLPENYAAVRFTWSCMLPPLFYTLTEISGIGLNWRKTRPIXXXXXXXXXX 366 + P+ ++ +YA+ V ML LF + ++ G ++T+ + Sbjct: 305 VLKPIVEKWSSDYASSWQYVPFFMLSMLFSSFSDFFGTNYIAAKQTKGVFMTSIYGTIV 364 Homology with a predicted ORF from N.meninsitidis (strain Al ORFIO shows 95.4% identity over a 475aa overlap with an ORF (ORF 10a) from strain A of N. meningitidis: 10 20 30 40 50 60 orf10. pep MDTKEILXYAAGSIGSAVLAVIILPLLSWYFPADDIGRIVLMQTAAGLTVSVLCLGLDQA I II I I I I II II I I I I II I I I II I I II II II II I I I I II II I II II I II II I II I II II I orf10a MDTKEILGYAAGSIGSAVLAVIILPLLSWYFPADDIGRIVLMQTAAGLTVSVLCLGLDQA 10 20 30 40 50 60 70 80 90 100 110 120 orf10. pep YVREYYATADKDTLFKTLFLPPLLSAAAIAALLLSRPSLPSEILFSLDDAAAGIGLVLFE II I I II I : I I I I II I II I I II II I II I II I I II I II I II II I II I I I II I II I I I I II I I orf10a YVREYYAAADKDTLFKTLFLPPLLSAAAIAALLLSRPSLPSEILFSLDDAAAGIGLVLFE 70 80 90 100 110 120 130 140 150 160 170 180 orf10. pep LSFLPIRFLLLVLRMEGRALAFSSAQLVPKLAILLLXPLTVGLLHFPANTAVLTAVYALA I I II I II I I II I I I I I I I I I I I I II I I I I I I I I II II I II II I I I II II I II I I I II I orf10a LSFLPIRFLLLVLRMEGRALAFSSAQLVSKLAILLLLPLTVGLLHFPANTAVLTAVYALA 130 140 150 160 170 180 190 200 210 220 230 240 orf10. pep NLAAAAFLLFQNRCRLKAVRHAPFSPAVLHRGXRYGIPIALSSIAYWGLASADRLFLKKY I I II I I II I I I I I I I II II I : I I I I I II II I II I I II I I II I I I II I I I II II II II I orf10a NLAAAAFLLFQNRCRLKAVRRAPFSSAVLHRGLRYGIPIALSSIAYWGLASADRLFLKKY 190 200 210 220 230 240 250 260 270 280 290 300 orf10.pep AGLEQLGVYSMGISFGGAALLFQΞIFSTVWTPYIFRAIEENAPPARLSATAESAAALLAS I I I I I I I I I I II I I II I I II I I II I I II I I I II I II II I II I II II II II I II I I I I II orf10a AGLEQLGVYSMGISFGGAALLFQSIFSTVWTPYIFRAIEANAPPARLSATAESAAALLAS 250 260 270 280 290 300 310 320 330 340 350 360 orf 10 . pep ALCXTGIFSPLASLLLPENYAAVRFIWSCMXPPLFCTLAEISGIGLNWRKTRPIALAT I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I : I I I I I I I I I I I I I I I I I I I I orf 10a ALCLTGIFSPLASLLLPENYAAVRFIVVSCMLPPLFCTLVEISGIGLNWRKTRPIALAT 310 320 330 340 350 360 370 380 390 400 410 419 orf10. pep LGALAANLLLLGLDRAVPAR-PXGAAVACAASFWLFFAFKTESSCRLWQPLKRLPLYLHT I I II I I I I I II I I III: I II I I I II I II I II : I I I II I I I I I I II II I II I : II orf10a LGALAANLLLLGL--AVPSGGARGAAVACAASFWLFFVFKTESSCRLWQPLKRLPLYMHT 370 380 390 400 410 420 430 440 450 460 470 orf10.pep LFCLTSSAAYTCFGTPANYPLFAGVWAAYLAGCILRHRKDLHKLFHYLKKQGFPLX I I II : I I II I I II I I I I I II I I II I I I : I I II I I II II II I I I II I I I I I II II I I orf10a LFCLASSAAYTCFGTPANYPLFAGVWAVYLAGCILRHRKDLHKLFHYLKKQGFPLX 420 430 440 450 460 470 The complete length ORF 10a nucleotide sequence <SEQ LD 377> is: 1 ATGGACACAA AAGAAATCCT CGGCTACGCG GCAGGCTCGA TCGGCAGCGC 51 GGTTTTAGCC GTCATCATCC TGCCGCTGCT GTCGTGGTAT TTCCCTGCCG 101 ACGACATCGG ACGCATCGTG CTGATGCAGA CGGCGGCGGG GCTGACGGTG 151 TCGGTGTTGT GCCTCGGGCT GGATCAGGCA TACGTCCGCG AATACTATGC 201 CGCCGCCGAC AAAGACACTT TGTTCAAAAC CCTGTTCCTG CCGCCGCTGC 251 TGTCTGCCGC CGCGATAGCC GCCCTGCTGC TTTCCCGCCC ATCCCTGCCG 301 TCTGAAATCC TGTTTTCGCT CGACGATGCC GCCGCCGGCA TCGGGCTGGT 351 GCTGTTTGAA CTGAGCTTCC TGCCCATCCG CTTTCTCTTA CTGGTTTTGC 401 GTATGGAAGG ACGCGCCCTT GCCTTTTCGT CCGCGCAACT CGTGTCCAAG 451 CTCGCCATCC TGCTGCTGCT GCCGCTGACG GTCGGGCTGC TGCACTTTCC 501 GGCGAACACC GCCGTCCTGA CCGCCGTTTA CGCGCTGGCA AACCTTGCCG 551 CCGCCGCCTT TTTGCTGTTT CAAAACCGAT GCCGTCTGAA GGCCGTCCGG 601 CGCGCACCGT TTTCATCCGC CGTCCTGCAT CGCGGCCTGC GCTACGGCAT 651 ACCGATCGCA CTAAGCAGCA TCGCCTATTG GGGGCTGGCA TCCGCCGACC 701 GTTTGTTCCT GAAAAAATAT GCCGGCCTAG AACAGCTCGG CGTTTATTCG 751 ATGGGTATTT CGTTCGGCGG AGCGGCATTA TTGTTCCAAA GCATCTTTTC 801 AACGGTCTGG ACACCGTATA TTTTCCGCGC AATCGAAGCA AACGCCCCGC 851 CCGCCCGCCT CTCGGCAACG GCAGAATCCG CCGCCGCCCT GCTTGCCTCC 901 GCCCTCTGCC TGACCGGCAT TTTCTCGCCC CTCGCCTCCC TCCTGCTGCC 951 GGAAAACTAC GCCGCCGTCC GGTTTATCGT CGTATCGTGT ATGCTGCCTC 1001 CGCTGTTTTG CACGCTGGTA GAAATCAGCG GCATCGGTTT GAACGTCGTC 1051 CGAAAAACAC GCCCGATCGC GCTCGCCACC TTGGGCGCGC TGGCGGCAAA 1101 CCTGCTGCTG CTGGGGCTTG CCGTACCGTC CGGCGGCGCG CGCGGCGCGG 1151 CGGTTGCCTG TGCCGCCTCA TTTTGGCTGT TTTTTGTTTT CAAGACCGAA 1201 AGCTCCTGCC GCCTGTGGCA GCCGCTCAAA CGCCTGCCGC TTTATATGCA 1251 CACATTGTTC TGCCTGGCCT CCTCGGCGGC CTACACCTGC TTCGGCACTC 1301 CGGCAAACTA CCCCCTGTTT GCCGGCGTAT GGGCGGTATA TCTGGCAGGC 1351 TGCATCCTGC GCCACCGGAA AGATTTGCAC AAACTGTTTC ATTATTTGAA 1401 AAAACAAGGT TTCCCATTAT GA This encodes a protein having amino acid sequence <SEQ ID 378>: 1 MDTKEILGYA AGSIGSAVLA VIILPLLSWY FPADDIGRIV LMQTAAGLTV 51 SVLCLGLDQA YVREYYAAAD KDTLFKTLFL PPLLSAAAIA ALLLSRPSLP 101 SEILFSLDDA AAGIGLVLFE LSFLPIRFLL LVLRMEGRAL AFSSAQLVSK 151 LAILLLLPLT VGLLHFPANT AVLTAVYALA NLAAAAFLLF QNRCRLKAVR 201 RAPFSSAVLH RGLRYGIPIA LSSIAYWGLA SADRLFLKKY AGLEQLGVYS 251 MGISFGGAAL LFQSIFSTVW TPYIFRAIEA NAPPARLSAT AESAAALLAS 301 ALCLTGIFSP LASLLLPENY AAVRFIWSC MLPPLFCTLV EISGIGLNW 351 RKTRPIALAT LGALAANLLL LGLAVPSGGA RGAAVACAAS FWLFFVFKTE 401 SSCRLWQPLK RLPLYMHTLF CLASSAAYTC FGTPANYPLF AGVWAVYLAG 451 CILRHRKDLH KLFHYLKKQG FPL* ORF 10a and ORF 10-1 show 95.4% identity in 475 aa overlap: 10 20 30 40 50 60 orf10-1. pep MDTKEILXYAAGSIGSAVLAVIILPLLSWYFPADDIGRIVLMQTAAGLTVSVLCLGLDQA I I I I I II I I I I M I I I I I II I I I I I I I I II I I II I I I II I I I I I I II I II I I II II I I I orf10a MDTKEILGYAAGSIGSAVLAVIILPLLSWYFPADDIGRIVLMQTAAGLTVSVLCLGLDQA 10 20 30 40 50 60 70 80 90 100 110 120 orf10-1. pep YVREYYATADKDTLFKTLFLPPLLSAAAIAALLLSRPSLPSEILFSLDDAAAGIGLVLFE I II II I I : I II I I II II II II II I I I I II II I I I I I I II I II I I I I I I II II I I II I I I I orf10a YVREYYAAADKDTLFKTLFLPPLLSAAAIAALLLΞRPSLPSEILFSLDDAAAGIGLVLFE 70 80 90 100 110 120 130 140 150 160 170 180 orf10-1. pep LSFLPIRFLLLVLRMEGRALAFSSAQLVPKLAILLLXPLTVGLLHFPANTAVLTAVYALA II I I I I II I I I I I I I I II II I I I II II I I I I I I I I I I I II I I I I I I II II I I II I I II orf10a LSFLPIRFLLLVLRMEGRALAFSSAQLVSKLAILLLLPLTVGLLHFPANTAVLTAVYALA 130 140 150 160 170 180 190 200 210 220 230 240 orf10-1. pep NLAAAAFLLFQNRCRLKAVRHAPFSPAVLHRGXRYGIPIALSSIAYWGLASADRLFLKKY I I II II II I I II I I I I II II : I I I I I II II I I I I I II I I I II I I I I I I I I I I I I II I I orf10a NLAAAAFLLFQNRCRLKAVRRAPFSSAVLHRGLRYGIPIALSSIAYWGLASADRLFLKKY 190 200 210 220 230 240 250 260 270 280 290 300 orf10-1.pep AGLEQLGVYSMGISFGGAALLFQSIFSTVWTPYIFRAIEENAPPARLSATAESAAALLAS II I I I I I I I II I I I I I I I I I II I II II I II I I I I I II II II I I I I I I I I I I I I I I I II I orf10a AGLEQLGVYSMGISFGGAALLFQSIFSTVWTPYIFRAIEANAPPARLSATAESAAALLAS 250 260 270 280 290 300 310 320 330 340 350 360 orf10-1. pep ALCXTGIFSPLASLLLPENYAAVRFIWSCMXPPLFCTLAEISGIGLNWRKTRPIALAT III II II I I I II I I II I I I II II I I I I I II II I I I I I : I I II II I I I II I I I I I I I I I orf10a ALCLTGIFSPLASLLLPENYAAVRFIVVSCMLPPLFCTLVEISGIGLNWRKTRPIALAT 310 320 330 340 350 360 370 380 390 400 410 419 orf10-1. pep LGALAANLLLLGLDRAVPAR-PXGAAVACAASFWLFFAFKTESSCRLWQPLKRLPLYLHT II I I I II I I I I I I III: I II II II I II II I I : II II I I I II I I I I I I I I I I : I I orf10a LGALAANLLLLGL—AVPSGGARGAAVACAASFWLFFVFKTESSCRLWQPLKRLPLYMHT 370 380 390 400 410 420 430 440 450 460 470 orf10-1. pep LFCLTSSAAYTCFGTPANYPLFAGVWAAYLAGCILRHRKDLHKLFHYLKKQGFPLX I I II : I II I I I I I II I II II I I I I I I I : I I II I I II I I I I I II II I I II I I II I I I orf10a LFCLASSAAYTCFGTPANYPLFAGVWAVYLAGCILRHRKDLHKLFHYLKKQGFPLX 420 430 440 450 460 470 Homology with a predicted ORF from N.sonorrhoeae ORFIO shows 94.1% identity over a 475aa overlap with a predicted ORF (ORFlO.ng) from N gonorrhoeae: orflOng.pep MDTKEILGYAAGSIGSAVLAVIILPLLSWYFPADDIGRIVLMQTAAGLTVSVLCLGLDQA 60 I I II I I I I I I I I I I I I II I I I I I I I I II II I II I I II II II I II II II II I I I II I I I I orflOnm MDTKEILXYAAGSIGSAVLAVIILPLLSWYFPADDIGRIVLMQTAAGLTVSVLCLGLDQA 60 orflOng.pep YVREYYAAADKDTLFKTLFLPPLLFSAAIAALLLSRPSLPSEILFSLDDAAAGIGLVLFE 120 II II I I I: I I II I I I II I II I II I : II I I II I I I I I I I I II I I I I I I I I I I II I I II I I orflOnm YVREYYATADKDTLFKTLFLPPLLSAAAIAALLLSRPSLPSEILFSLDDAAAGIGLVLFE 120 orflOng . pep LSFLPIRFLLLVLRMEGRALAFSSAQLVPKLAILLLLPLTVGLLHFPAΝTSVLTAVYALA 180 II I I II II II I II I II I II II I II I II II II I II I I I II I I II I II II I : II I I I I I I I orflOnm LSFLPIRFLLLVLRMEGRALAFSSAQLVPKLAILLLXPLTVGLLHFPAΝTAVLTAVYALA 180 orflOng.pep ΝLAAAAFLLFQΝRCRLKAVRRAPFSPAVLHRGLRYGIPLALSSLAYWGLASADRLFLKKY 240 I II I I I II I II I II I II II I : II I I I II I I II I I I I I : I II I : I I I I I I I I II I I II II orflOnm ΝLAAAAFLLFQΝRCRLKAVRHAPFSPAVLHRGXRYGIPIALSSIAYWGLASADRLFLKKY 240 orflOng.pep AGLEQLGVYSMGISFGGAALLLQSIFSTVWTPYIFRAIEEΝATPARLSATAESAAALLAS 300 I I I I I I I II I II I II I II II I : I I II I II II I II I II I II I I II II II I I II I I I I I I I orflOnm AGLEQLGVYSMGISFGGAALLFQSIFSTVWTPYIFRAIEEΝAPPARLSATAESAAALLAS 300 orflOng.pep ALCLTGIFSPLASLLLPEΝYAAVRFTWSCMLPPLFYTLTEISGIGLΝVVRKTRPIALAT 360 III I II I II I I I I II I I I I I I II I I II II I I I I I I: II I I II I I I I II II I I II II orflOnm ALCXTGIFSPLASLLLPEΝYAAVRFIWSCMXPPLFCTLAEISGIGLΝWRKTRPIALAT 360 370 380 390 400 410 orflOng . pep LGALAANLLLLGL—AVPSGGTRGAAVACAASFWLFFVFKTESSCRLWQPLKRLPLYMHT I I II I II I I I I I I III: I I II I I I I I II I I I : I I I I II I I II I I I I I II I I : II orflOnm LGALAANLLLLGLDRAVPAR-PXGAAVACAASFWLFFAFKTESSCRLWQPLKRLPLYLHT 370 380 390 400 410 420 430 440 450 460 470 orflOng . pep LFCLASSAAYTCFGTPANYPLFAGVWAAYLAGCILRHRKNLHKLFHYLKKQGFPLX I I II : II I I I II II I I I I II I I I I II II II II I II II II: I II I I I I I I II II I I I orflOnm LFCLTSSAAYTCFGTPANYPLFAGVWAAYLAGCILRHRKDLHKLFHYLKKQGFPLX 420 430 440 450 460 470 The complete length ORFlOng nucleotide sequence <SEQ ID 379> is: 1 ATGGACACAA AAGAAATCCT CGGCTACGCG GCAGGCTCGA TCGGCAGCGC 51 GGTTTTAGCC GTCATCATCC TGCCGCTGCT GTCGTGGTAT TTCcccgCCG 101 ACGACATCGG GCGCATCGTG CTGATGCAGA CGGCGGCGGG ACTGACGGTG 151 TCGGTATTGT GCCTCGGGCT GGATCAGGCA TACGTCCGCG AATACTATGC 201 CGCCGCCGAC AAAGACACTT TGTTCAAAAC CCTGTTCCTG CCGCCGCTGC 251 TGTTTTCCGC CGCGATAGCC GCCCTGCTGC TTTCCCGCCC GTCCCTGCCG 301 TCTGAAATCC TGTTTTCGCT CGACGATGCC GCCGCCGGCA TCGGGCTGGT 351 GCTGTTTGAA CTGAGCTTCC TGCCCATCCG CTTTCTCTTA CTGGTTTTGC 401 GTATGGAAGG GCGCGCCCTT GCCTTTTCGT CCGCGCAACT CGTGCCCAAA 451 CTCGCCATTC TGCTGCTGTT GCCGCTGACG GTCGGGCTGC TGCACTTTCC 501 GGCGAACACC TCCGTCCTGA CCGCCGTTTA CGCGCTGGCA AACCTTGCCG 551 CCGCCGCCTT TTTGCTGTTT CAAAACCGAT GCCGTCTGAA GGCCGTCCGG 601 CGCGCGCCGT TTTCGCCCGC CGTCCTGCAC CGGGGGCTGC GCTACGGCAT 651 ACCGCTCGCA CTGAGCAGCC TTGCCTATTG GGGGCTGGCA TCCGCCGACC 701 GTTTGTTCCT GAAAAAATAT GCGGGCCTGG AACAGCTCGG CGTTTATTCG 751 ATGGGTATTT CGTTCGGCGG GGCGGCATTA TTGCTCCAAA GCATCTTTTC 801 AACGGTCTGG ACACCGTATA TTTTCCGTGC AATCGAAGAA AACGCCACGC 851 CCGCCCGCCT CTCGGCAACG GCAGAATCCG CCGCCGCCCT GCTTGCCTCC 901 GCCCTCTGCC TGACCGGAAT TTTCTCGCCC CTCGCCTCCC TCCTGCTGCC 951 GGAAAACTAC GCCGCCGTCC GGTTTACCGT CGTATCGTGT ATGCTGccgc 1001 cgctGTTTTA CACGCTGACC GAAATCAGCG GCATCGGTTT GAACGTCGTC 1051 CGCAAAACGC GTCCGATCGC GCTTGCCACC TTGGGCGCGC TGGCGGCAAA 1101 CCTGCTGCTG CTGGGGCTTG CCGTACCGTC CGGCGGCACG CGCGGCGCGG 1151 CGGTTGCCTG TGCCGCCTCA TTCTGGTTGT TTTTTGTTTT CAAGACAGAA 1201 AGCTCCTGCC GCCTGTGGCA GCCGCTCAAA CGCCTGCCGC TTTATATGCA 1251 CACATTGTTC TGCCTgGCCT CCTCGGCGGC CTACACCTGC TTCGGCACAC 1301 CGGCAAACTA CCCcctgttt gccggcgtAT GGGCGGCATA TCTGGCAGGC 1351 TGCATCCTGC GCCACCGGAA AAATTTGCAC AAACTGTTTC ATTATTTGAA 1401 AAAACAAGGT TTCCCATTAT GA This encodes a protein having amino acid sequence <SEQ ID 380>: 1 MDTKEILGYA AGSIGSAVLA VIILPLLSWY FPADDIGRIV LMQTAAGLTV 51 SVLCLGLDQA YVREYYAAAD KDTLFKTLFL PPLLFSAAIA ALLLSRPSLP 101 SEILFSLDDA AAGIGLVLFE LSFLPIRFLL LVLRMEGRAL AFSSAQLVPK 151 LAILLLLPLT VGLLHFPANT SVLTAVYALA NLAAAAFLLF QNRCRLKAVR 201 RAPFSPAVLH RGLRYGIPLA LSSIAYWGLA SADRLFLKKY AGLEQLGVYS 251 MGISFGGAAL LLQSIFSTVW TPYIFRAIEE NATPARLSAT AESAAALLAS 301 ALCLTGIFSP LASLLLPENY AAVRFTWSC MLPPLFYTLT EISGIGLNW 351 RKTRPIALAT LGALAANLLL LGLAVPSGGT RGAAVACAAS FWLFFVFKTE 401 SSCRLWQPLK RLPLYMHTLF CLASSAAYTC FGTPANYPLF AGVWAAYLAG 451 CILRHRKNLH KLFHYLKKQG FPL* ORFlOng and ORF 10-1 show 96.4% identity in 473 aa overlap: 10 20 30 40 50 60 orf10-1. pep MDTKEILGYAAGSIGSAVLAVIILPLLSWYFPADDIGRIVLMQTAAGLTVSVLCLGLDQA I II I I I I I I I I II I II I II I I I I I I I I I I II II I I I I II II II I I I I I I I II I I II II I I orflOng-1 MDTKEILGYAAGSIGSAVLAVIILPLLSWYFPADDIGRIVLMQTAAGLTVSVLCLGLDQA 10 20 30 40 50 60 70 80 90 100 110 120 orf10-1. pep YVREYYATADKDTLFKTLFLPPLLSAAAIAALLLSRPSLPSEILFSLDDAAAGIGLVLFE I II I I I I: I I II I I I II I I I I I I I : I II II I II I II I I I I II II I II I II I II I I I II I orflOng-1 YVREYYAAADKDTLFKTLFLPPLLFSAAIAALLLSRPSLPSEILFSLDDAAAGIGLVLFE 70 80 90 100 110 120 130 140 150 160 170 180 orf10-1. pep LSFLPIRFLLLVLRMEGRALAFSSAQLVPKLAILLLLPLTVGLLHFPANTAVLTAVYALA II II I II II I II I I I II II II II II II II II II I II I I I I II I I I I I I I I : II I I II I I I orflOng-1 LSFLPIRFLLLVLRMEGRALAFSSAQLVPKLAILLLLPLTVGLLHFPANTSVLTAVYALA 130 140 150 160 170 180 190 200 210 220 230 240 orf10-1. pep NLAAAAFLLFQNRCRLKAVRHAPFSPAVLHRGLRYGIPIALSSIAYWGLASADRLFLKKY I I II II I II II I I I I I I I I I : I II II II II II I II II I : I II I : I I II I I II I I II I I I I orflOng-l NLAAAAFLLFQNRCRLKAVRRAPFSPAVLHRGLRYGIPLALSSLAYWGLASADRLFLKKY 190 200 210 220 230 240 250 260 270 280 290 300 orf10-1.pep AGLEQLGVYSMGISFGGAALLFQSIFSTVWTPYIFRAIEENAPPARLSATAESAAALLAS II I II II II I I I II I I I I II I : I II II II II I II II I I I I I I I I I I I II II I I II I I II orflOng-l AGLEQLGVYSMGISFGGAALLLQSIFSTVWTPYIFRAIEENATPARLSATAESAAALLAS 250 260 270 280 290 300 310 320 330 340 350 360 orf10-1. pep ALCLTGIFSPLASLLLPENYAAVRFIWSCMLPPLFCTLAEISGIGLNVVRKTRPIALAT I I I I I I I II I I I II I I I I I I I II II I II I II II I I I I I I II I I I I II II I I II II I orflOng-l ALCLTGIFSPLASLLLPENYAAVRFTWSCMLPPLFYTLTEISGIGLNWRKTRPIALAT 310 320 330 340 350 360 370 380 390 400 410 420 orf10-1. pep LGALAANLLLLGLAVPSGGARGAAVACAASFWLFFAFKTESSCRLWQPLKRLPLYLHTLF II I II II I I I I I I I II I I I : I I II I I I I I I I II I I : II I II I I I I I II I I II I I M I I I I orflOng-l LGALAANLLLLGLAVPSGGTRGAAVACAASFWLFFVFKTESSCRLWQPLKRLPLYMHTLF 370 380 390 400 410 420 430 440 450 460 470 orf10-1. pep CLTSSAAYTCFGTPANYPLFAGVWAAYLAGCILRHRKDLHKLFHYLKKQGFPLX II: II I II II I I I I II I I I I I I I I I II II II I II I II: I I I I I II I I I I II II I orflOng-l CLASSAAYTCFGTPANYPLFAGVWAAYLAGCILRHRKNLHKLFHYLKKQGFPLX 430 440 450 460 470 Based on this analysis, including the presence of a putative leader peptide and several transmembrane segments and the presence of a leucine-zipper motif (4 Leu residues spaced by 6 aa, shown in bold), it is predicted that these proteins from N. meningitidis and N.gonorrhoeae, and their epitopes, could be useful antigens for vaccines or diagnostics, or for raising antibodies. Example 45 The following partial DNA sequence was identified in N. meningitidis <SEQ ID 381>: 1.. ATCCTGAAAC CGCATAACCA GCTTAAGGAA GACATCCAAC CTGATCCGGC 51 CGATCAAAAC GCCTTGTCCG AACCGGATGC TGCGACAGAG GCAGAGCAGT 101 CGGATGCGGA AAATGCTGCC GACAAGCAGC CCGTTGCCGA TAAAGCCGAC 151 GAGGTTGAAG AAAAGGCGGG CGAGCCGGAA CGGGAAGAGC CGGACGGACA 201 GGCAGTGCGT AAGAAAGCGC TGACGGAAGA GCGTGAACAA ACCGTCAGGG 251 AAAAAGCGCA GAAGAAAGAT GCCGAAACGG TTAAAATACA AGCGGTAAAA 301 CCGTCTAAAG AAACAGAGAA AAAAGCTTCA AAAGAAGAGA AAAAGGCGGC 351 GAAGGAAAAA GTTGCACCCA AACCAACCCC GGAACAAATC CTCAACAGCG 401 GCAgCATCGA AAAmGCGCGC AgTGCCGCCG CCAAAGAAGT GCAGAAAATG 451 AA.AACGTCC GACAAGGCGG AAGC.AACGC ATTATCTGCA AATGGGCGCG 501 TATGCCGACC GTCAGAGCGC GGAAGGGCAG CGTGCCAAAC TGGCAATCTT 551 GGGCATATCT TCCAAGGTGG TCGGTTATCA GGCGGGACAT AAAACGCTTT 601 ACCGGGTGCA AAGCGGCAAT ATGTCTGCCG ATGCGGTGA This coπesponds to the amino acid sequence <SEQ ID 382; ORF65>: 1.. ILKPHNQLKE DIQPDPADQN ALSEPDAATE AEQSDAENAA DKQPVADKAD 51 EVEEKAGEPE REEPDGQAVR KKALTEEREQ TVREKAQKKD AETVKIQAVK 101 PSKETEKKAS KEEKKAAKEK VAPKPTPEQI LNSGSIEXAR SAAAKEVQKM 151 XNVRQGGSXR IICKWARMPT VRARKGSVPN WQSWAYLPRW SVIRRDIKRF 201 TGCKAAICLP MR* Further work revealed the complete nucleotide sequence <SEQ ID 383>: 1 ATGTTTATGA ACAAATTTTC CCAATCCGGA AAAGGTCTGT CCGGTTTTTT 51 CTTCGGTTTG ATACTGGCGA CGGTCATTAT TGCCGGTATT TTGTTTTATC 101 TGAACCAGAG CGGTCAAAAT GCGTTCAAAA TCCCGGCTTC GTCGAAGCAG 151 CCTGCAGAAA CGGAAATCCT GAAACCGAAA AACCAGCCTA AGGAAGACAT 201 CCAACCTGAA CCGGCCGATC AAAACGCCTT GTCCGAACCG GATGCTGCGA 251 CAGAGGCAGA GCAGTCGGAT GCGGAAAAAG CTGCCGACAA GCAGCCCGTT 301 GCCGATAAAG CCGACGAGGT TGAAGAAAAG GCGGGCGAGC CGGAACGGGA 351 AGAGCCGGAC GGACAGGCAG TGCGTAAGAA AGCGCTGACG GAAGAGCGTG 401 AACAAACCGT CAGGGAAAAA GCGCAGAAGA AAGATGCCGA AACGGTTAAA 451 AAACAAGCGG TAAAACCGTC TAAAGAAACA GAGAAAAAAG CTTCAAAAGA 501 AGAGAAAAAG GCGGCGAAGG AAAAAGTTGC ACCCAAACCA ACCCCGGAAC 551 AAATCCTCAA CAGCGGCAGC ATCGAAAAAG CGCGCAGTGC CGCCGCCAAA 601 GAAGTGCAGA AAATGAAAAC GTCCGACAAG GCGGAAGCAA CGCATTATCT 651 GCAAATGGGC GCGTATGCCG ACCGTCAGAG CGCGGAAGGG CAGCGTGCCA 701 AACTGGCAAT CTTGGGCATA TCTTCCAAGG TGGTCGGTTA TCAGGCGGGA 751 CATAAAACGC TTTACCGGGT GCAAAGCGGC AATATGTCTG CCGATGCGGT 801 GAAAAAAATG CAGGACGAGT TGAAAAAACA TGAAGTCGCC AGCCTGATCC 851 GTTCTATCGA AAGCAAATAA This corresponds to the amino acid sequence <SEQ ID 384; ORF65-l>: 1 MFMNKFSQSG KGLSGFFFGL ILATVIIAGI LFYLNQSGQN AFKIPASSKQ 51 PAETEILKPK NQPKEDIQPE PADQNALSEP DAATEAEQSD AEKAADKQPV 101 ADKADEVEEK AGEPEREEPD GQAVRKKALT EEREQTVREK AQKKDAETVK 151 KQAVKPSKET EKKASKEEKK AAKEKVAPKP TPEQILNSGS IEKARSAAAK 201 EVQKMKTSDK AEATHYLQMG AYADRQSAEG QRAKLAILGI SSKWGYQAG 251 HKTLYRVQSG NMSADAVKKM QDELKKHEVA SLIRSIESK* Computer analysis of this amino acid sequence gave the following results: Homology with a predicted ORF from N.meninsitidis (strain A) ORF65 shows 92.0% identity over a 150aa overlap with an ORF (ORF65a) from sfrain A of N meningitidis: 10 20 30 orf65. pep ILKPHΝQLKEDIQPDPADQΝALSEPDAATE I I I I : I I I I II I I : I I I II I II II I II I orf65a IIAGILFYLΝQSGQΝAFKIPVPSKQPAETEILKPKΝQPKEDIQPEPADQΝALSEPDAAKE 30 40 50 60 70 80 40 50 60 70 80 90 orf65.pep AEQSDAEΝAADKQPVADKADEVEEKAGEPEREEPDGQAVRKKALTEEREQTVREKAQKKD I I I II I I: I I II I I I I I I I II I I I I I I I II I: I I I I I I II I I I I I I II II I I I I I I I orf65a AEQSDAEKAADKQPVADKADEVEEKADEPEREKSDGQAVRKKALTEEREQTVGEKAQKKD 90 100 110 120 130 140 100 110 120 130 140 150 orf65. pep AETVKIQAVKPSKETEKKASKEEKKAAKEKVAPKPTPEQILΝSGSIEXARSAAAKEVQKM II I I I II I I I II II II I II I I II I I I II I II I II I I II I II II II I I I I I I I I I I I I orf65a AETVKKQAVKPSKETEKKASKEEKKAEKEKVAPKPTPEQILΝSGSIEKARSAAAKEVQKM 150 160 170 180 190 200 160 170 180 190 200 210 orf65. pep XΝVRQGGSXRIICKWARMPTVRARKGSVPΝWQSWAYLPRWSVIRRDIKRFTGCKAAICLP orf65a KTPDKAEATHYLQMGAYADRRSAEGQRAKLAILGISSKWGYQAGHKTLYRVQSGΝMSAD 210 220 230 240 250 260 The complete length ORF65a nucleotide sequence <SEQ D 385> is: 1 ATGTTTATGA ACAAATTTTC CCAATCCGGA AAAGGTCTGT CCGGTTTTTT 51 CTTCGGTTTG ATACTGGCGA CGGTCATTAT TGCCGGTATT TTGTTTTATC 101 TGAACCAGAG CGGTCAAAAT GCGTTCAAAA TCCCGGTTCC GTCGAAGCAG 151 CCTGCAGAAA CGGAAATCCT GAAACCGAAA AACCAGCCTA AGGAAGACAT 201 CCAACCTGAA CCGGCCGATC AAAACGCCTT GTCCGAACCG GATGCTGCGA 251 AAGAGGCAGA GCAGTCGGAT GCGGAAAAAG CTGCCGACAA GCAGCCCGTT 301 GCCGACAAAG CCGACGAGGT TGAGGAAAAG GCGGACGAGC CGGAGCGGGA 351 AAAGTCGGAC GGACAGGCAG TGCGCAAGAA AGCACTGACG GAAGAGCGTG 401 AACAAACCGT CGGGGAAAAA GCGCAGAAGA AAGATGCCGA AACGGTTAAA 451 AAACAAGCGG TAAAACCATC TAAAGAAACA GAGAAAAAAG CTTCAAAAGA 501 AGAGAAAAAG GCGGAGAAGG AAAAAGTTGC ACCCAAACCG ACCCCGGAAC 551 AAATCCTCAA CAGCGGCAGC ATCGAAAAAG CGCGCAGTGC CGCTGCCAAA 601 GAAGTGCAGA AAATGAAAAC GCCCGACAAG GCGGAAGCAA CGCATTATCT 651 GCAAATGGGC GCGTATGCCG ACCGCCGGAG CGCGGAAGGG CAGCGTGCCA 701 AACTGGCAAT CTTGGGCATA TCTTCCAAGG TGGTCGGTTA TCAGGCGGGA 751 CATAAAACGC TTTACCGGGT GCAAAGCGGC AATATGTCTG CCGATGCGGT 801 GAAAAAAATG CAGGACGAGT TGAAAAAACA TGAAGTCGCC AGCCTGATCC 851 GTTCTATCGA AAGCAAATAA This encodes a protein having amino acid sequence <SEQ LD 386>: 1 MFMNKFSQSG KGLSGFFFGL ILATVIIAGI LFYLNQSGQN AFKIPVPSKQ 51 PAETEILKPK NQPKEDIQPE PADQNALSEP DAAKEAEQSD AEKAADKQPV 101 ADKADEVEEK ADEPEREKSD GQAVRKKALT EEREQTVGEK AQKKDAETVK 151 KQAVKPSKET EKKASKEEKK AEKEKVAPKP TPEQILNSGS IEKARSAAAK 201 EVQKMKTPDK AEATHYLQMG AYADRRSAEG QRAKLAILGI SSKWGYQAG 251 HKTLYRVQSG NMSADAVKKM QDELKKHEVA SLIRSIESK* ORF65a and ORF65-1 show 96.5% identity in 289 aa overlap: 10 20 30 40 50 60 orf65a . pep MFMNKFSQSGKGLSGFFFGLILATVIIAGILFYLNQSGQNAFKIPVPSKQPAETEILKPK I I I I I I I I I I I II I I I I I I II I I I II I I II I I I I II I I I II I II I : I I I I I I I II II I I orf65-1 MFMNKFSQSGKGLSGFFFGLILATVIIAGILFYLNQSGQNAFKIPASSKQPAETEILKPK 10 20 30 40 50 60 70 80 90 100 110 120 orf65a . pep NQPKEDIQPEPADQNALSEPDAAKEAEQSDAEKAADKQPVADKADEVEEKADEPEREKSD II I I I I II I I I I I I I I I I I I I I I I I I II I I II II I II II I I I II I II I II Mill: I orf65-1 NQPKEDIQPEPADQNALSEPDAATEAEQSDAEKAADKQPVADKADEVEEKAGEPEREEPD 70 80 90 100 110 120 130 140 150 160 170 180 orf65a . pep GQAVRKKALTEEREQTVGEKAQKKDAETVKKQAVKPSKETEKKASKEEKKAEKEKVAPKP I I I I I I I I I I I I I I I I I I I I I II I II II II I II I I I I I I I I I I I I I II I I I II II II I orf65-1 GQAVRKKALTEEREQTVREKAQKKDAETVKKQAVKPSKETEKKASKEEKKAAKEKVAPKP 130 140 150 160 170 180 190 200 210 220 230 240 orf65a . pep TPEQILNSGSIEKARSAAAKEVQKMKTPDKAEATHYLQMGAYADRRSAEGQRAKLAILGI I I I II I II I I I I I I II I I I II I I II I I II I I I II I I II II I I I I : I I II I I I I II II II orf 65-1 TPEQILNSGSIEKARSAAAKEVQKMKTSDKAEATHYLQMGAYADRQSAEGQRAKLAILGI 190 200 210 220 230 240 250 260 270 280 290 orf 65a . pep SSKWGYQAGHKTLYRVQSGNMSADAVKKMQDELKKHEVASLIRSIESKX I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I orf 65-1 SSKWGYQAGHKTLYRVQSGNMSADAVKKMQDELKKHEVASLIRSIESKX 250 260 270 280 290 Homology with a predicted ORF from N.sonorrhoeae ORF65 shows 89.6% identity over a 212aa overlap with a predicted ORF (ORF65.ng) from N. gonorrhoeae: 30 40 50 60 70 80 0RF65ng IIAGILLYLNQGGQNAFKIPAPSKQPAETEILKLKNQPKEDIQPEPADQNALSEPDVAKE III l I I I I I I: I I I II I II M l I ORF65 ILKPHNQLKEDIQPDPADQNALSEPDAATE 10 20 30 90 100 110 120 130 140 ORF65ng AEQSDAEKAADKQPVADKADEVEEKAGEPEREEPDGQAVRKKALTEEREQTVREKAQKKD II I I II I : I II II I I I I II I I I I I I II I I I I I I I I I I I I I I I I I II II I I I I II I II II I ORF65 AEQSDAENAADKQPVADKADEVEEKAGEPEREEPDGQAVRKKALTEEREQTVREKAQKKD 40 50 60 70 80 90 150 160 170 180 190 200 ORF65ng AETVKKKAVKPSKETEKKASKEEKKAAKEKVAPKPTPEQILNSRSIEKARSAAAKEVQKM I II I I : II II I II II II I II II I II II II I I I I I II I I I II I III I I I I II II I I I I ORF65 AETVKIQAVKPSKETEKKASKEEKKAAKEKVAPKPTPEQILNSGSIEXARSAAAKEVQKM 100 110 120 130 140 150 210 220 230 240 250 260 ORF65ng KNFGQGGSQRIICKWARMPNPGARKGSVPNWQSWAYLPKWSAIRRDIKRFTACKAAICPP I I II I I I I I I I I I I I : II I I I I I II II II I I I : I I : I I I I I I I II : I II I I I I ORF65 XNVRQGGSXRIICKWARMPTVRARKGSVPNWQSWAYLPRWSVIRRDIKRFTGCKAAICLP 160 170 180 190 200 210 ORF65ng MR I I O URKF-6b5ό MMRK An ORF65ng nucleotide sequence <SEQ LD 387> was predicted to encode a protein having amino acid sequence <SEQ LD 388>: 1 MFMNKFSQSG KGLSGFFFGL ILATVIIAGI LLYLNQGGQN AFKIPAPSKQ 51 PAETEILKLK NQPKEDIQPE PADQNALSEP ' DVAKEAEQSD AEKAADKQPV 101 ADKADEVEEK AGEPEREEPD GQAVRKKALT EEREQTVREK AQKKDAETVK 151 KKAVKPSKET EKKASKEEKK AAKEKVAPKP TPEQILNSRS IEKARSAAAK 201 EVQKMKNFGQ GGSQRIICKW ARMPNPGARK GSVPNWQSWA YLPKWSAIRR 251 DIKRFTACKA AICPPMR* After further analysis, the complete gonococcal DNA sequence <SEQ ID 389> was found to be: 1 ATGTTTATGA ACAAATTTTC CCAATCCGGA AAAGGTCTGT CCGGTTTCTT 51 CTTCGGTTTG ATACTGGCAA CGGTCATTAT TGCCGGTATT TTGCTTTATC 101 TGAACCAGGG CGGTCAAAAT GCGTTCAAAA TCCCGGCTCC GTCGAAGCAG 151 CCTGCAGAAA CGGAAATCCT GAAACTGAAA AACCAGCCTA AGGAAGACAT 201 CCAACCTGAA CCGGCCGATC AAAACGCCTT GTCCGAACCG GATGTTGCGA 251 AAGAGGCAGA GCAGTCGGAT GCGGAAAAAG CTGCCGACAA GCAGCCCGTT 301 GCCGACAAag ccgacgAGGT TGAAGAAAag GcGGgcgAgc cggaACGGga 351 aGAGCCGGAC ggACAGGCAG TGCGCAAGAA AGCACTGAcg gAAGAgcGTG 401 AACAAACcgt cagggAAAAA GCGCagaaga AAGATGCCGA AACGgTTAAA 451 AAacaaGCgg tAaaaccgtc tAAAGAAACa gagaaaaaag cTtcaaaaga 501 agagaaaaag gcggcgaaag aaaAAGttgc acccaaaccg accccggaaC 551 aaatcctcaa cagccgCagc atcgaaaaag cgcgtagtgc cgctgccaaa 601 gaAgtgcaGA AAatgaaaaa ctTtgggcaa ggcgGaagcc aacgcattaT 651 CTGcaaatgg gcgcgtatgc cgaccgtccg gagcgcggaA gggcagcgtg 701 ccaaACtggc aAtcttgGgc atatctTccg aagtggtcgG CTATCAGGCG 751 GGACATAAAA CGCTTTACCG CGTGCAAagc GGCAatatgt ccgccgatgc 801 gGTGAAAAAA ATGCAGGACG AGTTGAAAAA GCATGGGGtt gcCAGCCTGA 851 TCCGTGcgAT TGAAGGCAAA TAA This encodes the following .amino acid sequence <SEQ D 390>: 1 MFMNKFSQSG KGLSGFFFGL ILATVIIAGI LLYLNQGGQN AFKIPAPSKQ 51 PAETEILKLK NQPKEDIQPE PADQNALSEP DVAKEAEQSD AEKAADKQPV 101 ADKADEVEEK AGEPEREEPD GQAVRKKALT EEREQTVREK AQKKDAETVK 151 KQAVKPSKET EKKASKEEKK AAKEKVAPKP TPEQILNSRS IEKARSAAAK 201 EVQKMKNFGQ GGSQRIICKW ARMPTVRSAE GQRAKLAILG ISSEWGYQA 251 GHKTLYRVQS GNMSADAVKK MQDELKKHGV ASLIRAIEGK ORF65ng-l and ORF65-1 show 89.0% identity in 290 aa overlap: 10 20 30 40 50 60 orf65-1. pep MFMNKFSQSGKGLSGFFFGLILATVIIAGILFYLNQSGQNAFKIPASSKQPAETEILKPK II I I I I I II I I I I I I I II II I I II II I I I IM I II I: I II II II II I I I II II I I I I I orf65ng-l MFMNKFSQSGKGLSGFFFGLILATVIIAGILLYLNQGGQNAFKIPAPSKQPAETEILKLK 10 20 30 40 50 60 70 80 90 100 110 120 orf65-1. pep NQPKEDIQPEPADQNALSEPDAATEAEQSDAEKAADKQPVADKADEVEEKAGEPEREEPD I II I II I I I II II I I I I I I MM I I II II I I I II I II I I I II I I I I I I I I I I I I II I I I orf65ng-l NQPKEDIQPEPADQNALSEPDVAKEAEQSDAEKAADKQPVADKADEVEEKAGEPEREEPD 70 80 90 100 110 120 130 140 150 160 170 180 orf65-1. pep GQAVRKKALTEEREQTVREKAQKKDAETVKKQAVKPSKETEKKASKEEKKAAKEKVAPKP II I I I II I II II II II I I I I I I I I I I I I I I I I I II I I I I I I I II I I I I I I I II I I II I I I orf65ng-l GQAVRKKALTEEREQTVREKAQKKDAETVKKQAVKPSKETEKKASKEEKKAAKEKVAPKP 130 140 150 160 170 180 190 200 210 220 230 239 orf65-1. pep TPEQILNSGSIEKARSAAAKEVQKMKTSDKAEATHYL-QMGAYADRQSAEGQRAKLAILG I I I I I I I I II I I I I I II I I I I II I I : :: : : : : : : : I I I I II I I I II I I orf65ng-l TPEQILNSRSIEKARSAAAKEVQKMKNFGQGGSQRIICKWARMPTVRSAEGQRAKLAILG 190 200 210 220 230 240 240 250 260 270 280 290 orf65-1.pep ISSKWGYQAGHKTLYRVQSGNMSADAVKKMQDELKKHEVASLIRSIESKX II I : I II I I I I II II II I I I I I I I I II II II I II I I I I I I I II I : I I : II orf65ng-l ISSEWGYQAGHKTLYRVQSGNMSADAVKKMQDELKKHGVASLIRAIEGKX 250 260 270 280 290 On this basis, including the presence of a putative transmembrane domain in the gonococcal protein, it is predicted that the proteins from Nmeningitidis and N.gonorrhoeae, and their epitopes, could be useful antigens for vaccines or diagnostics, or for raising antibodies. Example 46 The following DNA sequence, believed to be complete, was identified in N. meningitidis <SEQ ED 391>: 1 ATGAACCACG ACATCACTTT CCTCACCCTG TTCCTACTCG GTkTCTTCGG 51 CGGAAcGCAC TGCATCGGTA TGTGCGGCGG ATTAAGCAGC GcGTTTGs . s 101 TCCAACTCCC CCCGCATATC AACCGCTTTT GGCTGATCCT GCTGCTTAAC 151 ACAGGACGGG TAAGCAGCTA TACGGCAAtC GGCCTGATAC TCGGATTAAT 201 CGGACAGGTC GGCGTTTCAC TCGAcCAaAC CCGCGTCCTG CAGAATATTT 251 TATACACGGC CGCCAACCTC CTGCTGCTCT TTTTAGGCTT ATACTTGAGC 301 GGTATTTCTT CCTTGGCGGC AAAAATCGAG AAaATCGGCA AACCGATATG 351 GCGGAACCTG AACCCGATAC TCAACCGGCT GTTACCCATA AAATCCATAC 401 CCGCCTGCCT tGCGgTCGGA ATATTATGGG GCTGGCTGCC GTGCGGACTG 451 GTTTACAGCG CGTCGCTTTA CGCGCTGGGA AgCGGTAGTG CGGCAACGGG 501 CGGGTTATAT ATGCTTGCCT TTGCACTGGG TACGCTGCCC AATCTTtTAG 551 CAATCGGCAT TTTtTCCCTG CAACTGAAwA AAATCATGCA AAACCGATAT 601 ATCCGCCTGT GTACGGGATT ATCCGTATCA TTATGGGCAT TATGGAAACT 651 TGCCGTCCTG TGGCTGTAA This coπesponds to the amino acid sequence <SEQ LD 392; ORF103>: 1 MNHDITFLTL FLLGXFGGTH CIGMCGGLSS AFXXQLPPHI NRFWLILLLN 51 TGRVSSYTAI GLILGLIGQV GVSLDQTRVL QNILYTAANL LLLFLGLYLS 101 GISSLAAKIE KIGKPIWRNL NPILNRLLPI KSIPACLAVG ILWGWLPCGL 151 VYSASLYALG SGSAATGGLY MLAFALGTLP NLLAIGIFSL QLXKIMQNRY 201 IRLCTGLSVS LWALWKLAVL WL* Further work elaborated the DNA sequence <SEQ LD 393> as: 1 ATGAACCACG ACATCACTTT CCTCACCCTG TTCCTACTCG GTTTCTTCGG 51 CGGAACGCAC TGCATCGGTA TGTGCGGCGG ATTAAGCAGC GCGTTTGCGC 101 TCCAACTCCC CCCGCATATC AACCGCTTTT GGCTGATCCT GCTGCTTAAC 151 ACAGGACGGG TAAGCAGCTA TACGGCAATC GGCCTGATAC TCGGATTAAT 201 CGGACAGGTC GGCGTTTCAC TCGACCAAAC CCGCGTCCTG CAGAATATTT 251 TATACACGGC CGCCAACCTC CTGCTGCTCT TTTTAGGCTT ATACTTGAGC 301 GGTATTTCTT CCTTGGCGGC AAAAATCGAG AAAATCGGCA AACCGATATG 351 GCGGAACCTG AACCCGATAC TCAACCGGCT GTTACCCATA AAATCCATAC 401 CCGCCTGCCT TGCGGTCGGA ATATTATGGG GCTGGCTGCC GTGCGGACTG 451 GTTTACAGCG CGTCGCTTTA CGCGCTGGGA AGCGGTAGTG CGGCAACGGG 501 CGGGTTATAT ATGCTTGCCT TTGCACTGGG TACGCTGCCC AATCTTTTAG 551 CAATCGGCAT TTTTTCCCTG CAACTGAAAA AAATCATGCA AAACCGATAT 601 ATCCGCCTGT GTACGGGATT ATCCGTATCA TTATGGGCAT TATGGAAACT 651 TGCCGTCCTG TGGCTGTAA This coπesponds to the .amino acid sequence <SEQ ID 394; ORF103-1>: 1 MNHDITFLTL FLLGFFGGTH CIGMCGGLSS AFALQLPPHI NRFWLILLLN 51 TGRVSSYTAI GLILGLIGQV GVSLDQTRVL QNILYTAANL LLLFLGLYLS 101 GISSLAAKIE KIGKPIWRNL NPILNRLLPI KSIPACLAVG ILWGWLPCGL 151 VYSASLYALG SGSAATGGLY MLAFALGTLP NLLAIGIFSL QLKKIMQNRY 201 IRLCTGLSVS LWALWKLAVL WL* Computer analysis of this amino acid sequence gave the following results: Homology with a predicted ORF from N.meninsitidis (sfrain Al ORF103 shows 93.8% identity over a 222aa overlap with an ORF (ORF103a) from strain A of N. meningitidis: 10 20 30 40 50 60 orf103.pep MNHDITFLTLFLLGXFGGTHCIGMCGGLSSAFXXQLPPHINRFWLILLLNTGRVSSYTAI II I I I I II II II I II II II II I I II I I I I I II I II II I I I I II I II I II I I I II I orf103a MNXDITFLTLFLLGFFGGTHCIGMCGGLSSAFALQLPPHINRXWLILLLNTGRVSSYTAI 10 20 30 40 50 60 70 80 90 100 110 120 orf103.pep GLILGLIGQVGVSLDQTRVLQNILYTAANLLLLFLGLYLSGISSLAAKIEKIGKPIWRNL I I II I I I I I I I I I I I II I I I I II I I II II II II I II I I I I I I I I I I I II I II I II I I I I orf103a GLILGLIGQVGVSLDQTRVXQNILYTAANLLLLFLGLYLSGISSLAAKIEKIGKPIWRNL 70 80 90 100 110 120 130 140 150 160 170 180 orf103.pep NPILNRLLPIKSIPACLAVGILWGWLPCGLVYSASLYALGSGSAATGGLYMLAFALGTLP I I II I I I II I I I I I I I II II II I II I II II I I I II I II II I I I II I I I I I II II I I II II orf103a NPILNRLLPIKSIPACLAVGILWGWLPCGLVYSASLYALGSGSAATGGLYMLAFALGTLP 130 140 150 160 170 180 190 200 210 220 orf103. pep NLLAIGIFSLQLXKIMQNRYIRLCTGLSVSLWALWKLAVLWLX II II I II II I I II I II I I II I II I I I I I I I I II I II I I I II I orf103a NLXAIGIFSLQLXKIMQNRYIRLCTGLSVSLWALWKLAVLWLX 190 200 210 220 The complete length ORF103a nucleotide sequence <SEQ ID 395> is: 1 ATGAACCANG ACATCACTTT CCTCACCCTG TTCCTACTCG GTTTCTTCGG 51 CGGAACGCAC TGCATCGGTA TGTGCGGCGG ATTAAGCAGC GCGTTTGCGC 101 TCCAACTCCC CCCGCATATC AACCGCTTNT GGCTGATCCT GCTGCTTAAC 151 ACAGGACGGG TAAGCAGCTA TACGGCAATC GGCCTGATAC TCGGATTAAT 201 CGGACAGGTC GGCGTTTCAC TCGACCAAAC CCGCGTCNTG CAGAATATTT 251 TATACACGGC CGCCAACCTC CTGCTGCTCT TTTTAGGCTT ATACTTGAGC 301 GGTATTTCTT CCTTGGCGGC AAAAATCGAG AAAATCGGCA AACCGATATG 351 GCGGAACCTG AACCCGATAC TCAACCGGCT GTTACCCATA AAATCCATAC 401 CCGCCTGCCT TGCGGTCGGA ATATTATGGG GCTGGCTGCC GTGCGGACTA 451 GTTTACAGCG CGTCGCTTTA CGCGCTGGGA AGCGGTAGTG CGGCAACGGG 501 CGGGTTATAT ATGCTTGCCT TTGCACTGGG TACGCTGCCC AATCTTTNGG 551 CAATCGGCAT TTTTTCCCTG CAACTGNAAA AAATCATGCA AAACCGATAT 601 ATCCGCCTGT GTACGGGATT ATCCGTATCA TTATGGGCAT TATGGAAACT 651 TGCCGTCCTG TGGCTGTAA This encodes a protein having amino acid sequence <SEQ LD 396>: 1 MNXDITFLTL FLLGFFGGTH CIGMCGGLSS AFALQLPPHI NRXWLILLLN 51 TGRVSSYTAI GLILGLIGQV GVSLDQTRVX QNILYTAANL LLLFLGLYLS 101 GISSLAAKIE KIGKPIWRNL NPILNRLLPI KSIPACLAVG ILWGWLPCGL 151 VYSASLYALG SGSAATGGLY MLAFALGTLP NLXAIGIFSL QLXKIMQNRY 201 IRLCTGLSVS LWALWKLAVL WL* ORF103a and ORF103-1 show 97.7% identity in 222 aa overlap: 10 20 30 40 50 60 orflC3a.pep MNXDITFLTLFLLGFFGGTHCIGMCGGLSSAFALQLPPHINRXWLILLLNTGRVSSYTAI II I II II I I II I I II I I II I I I I I I II I II I II I I II I I I I I I I I I II II I I I II I I I orf103-1 MNHDITFLTLFLLGFFGGTHCIGMCGGLSSAFALQLPPHINRFWLILLLNTGRVSSYTAI 10 20 30 40 50 60 70 80 90 100 110 120 orf103a. pep GLILGLIGQVGVSLDQTRVXQNILYTAANLLLLFLGLYLSGISSLAAKIEKIGKPIWRNL I I II II II I I II II I I I I I I I II II I II I I II I I I I I I I I I I I I I I I I II I I I I II I II orf103-1 GLILGLIGQVGVSLDQTRVLQNILYTAANLLLLFLGLYLSGISSLAAKIEKIGKPIWRNL 70 80 90 100 110 120 130 140 150 160 170 180 orf103a . pep NPILNRLLPIKSIPACLAVGILWGWLPCGLVYSASLYALGSGSAATGGLYMLAFALGTLP I I II II II I I II I I I I I I I I II II I II II II I I I II I I I I I I I I II I I I I I II II I I I I I orf103-1 NPILNRLLPIKSIPACLAVGILWGWLPCGLVYSASLYALGSGSAATGGLYMLAFALGTLP 130 140 150 160 170 180 190 200 210 220 orf103a . pep NLXAIGIFSLQLXKIMQNRYIRLCTGLSVSLWALWKLAVLWLX II II I I I I I II I I I I I I I II I II I I II I I I II II II I I I II orf103-1 NLLAIGIFSLQLKKIMQNRYIRLCTGLSVSLWALWKLAVLWLX 190 200 210 220 Homology with a predicted ORF from N.sonorrhoeae ORF 103 shows 95.5% identity over a 222aa overlap with a predicted ORF (ORF103.ng) from N. gonorrhoeae: orf103.pep MNHDITFLTLFLLGXFGGTHCIGMCGGLSSAFXXQLPPHINRFWLILLLNTGRVSSYTAI 60 I I I I II I I I I I II I II I II I I I I I I II I II I I I I I I I I II I I I II I I II I: II II I I orfl03ng MNHDITFLTLFLLGFFGGTHCIGMCGGLSSAFALQLPPHINRFWLILLLNTGRISSYTAI 60 orf103.pep GLILGLIGQVGVSLDQTRVLQNILYTAANLLLLFLGLYLSGISSLAAKIEKIGKPIWRNL 120 I I II I I I : I: I I I I II I I I II I II I: I II II I II II II II I II II I I I I II I I I I II I orfl03ng GLMLGLIGQLGISLDQTRVLQNILYTASNLLLLFLGLYLSGISSLAAKIEKIGKPIWRNL 120 orf103.pep NPILNRLLPIKSIPACLAVGILWGWLPCGLVYSASLYALGSGSAATGGLYMLAFALGTLP 180 I I II I I I II I II II II I II II I I II I II I I II I I I I I I I I I II I: I II I I I I I I II I I I I orfl03ng NPILNRLLPIKSIPACLAVGILWGWLPCGLVYSASLYALGSGSATTGGLYMLAFALGTLP 180 orf103. pep NLLAIGIFSLQLXKIMQNRYIRLCTGLSVΞLWALWKLAVLWL 222 I I I I I I I I I II I I I I I I I I I I I I I I I I I I II I II I I I I I I I orfl03ng NLLAIGIFSLQLKKIMQNRYIRLCTGLSVSLWALWKLAVLWL 222 The complete length ORF103ng nucleotide sequence <SEQ ID 397> is: 1 ATGAACCACG ACATCACTTT CCTCACCCTG TTCCTGCTCG GTTTCTTCGG 51 CGGAACTCAC TGCATCGGTA TGTGCGGCGG ATTAAGCAGC GCGTTTGCGC 101 TCCAACTCCC CCCGCATATC AACCGCTTTT GGCTGATTCT GCTGCTTAAC 151 ACAGGACGGA TAAGCAGCTA TACGGCAATC GGCCTGATGC TCGGATTAAT 201 CGGACAACTC GGCATTTCAC TCGACCAAAc ccgcgTCCTG CAAAATATTT 251 tatacacagc ctccaaCCTC CTGCTGCTCT TTTTAGGCTT ATACTTGAGC 301 GGTATTTCTT CCTTGGCGGC AAAAATCGAG AAAATCGGCA AACCGATATG 351 GCGCAACCTG AACCCGATAC TCAACCGGCT GCTGCCCATA AAATCCATAC 401 CCGCCTGCCT TGCTGTCGGA ATATTATGGG GCTGGCTGCC GTGCGGACTG 451 GTTTACAGCG CATCACTTTA CGCGCTGGGA AGCGGTAGTG CGACAACCGG 501 CGGACTGTAT ATGCTTGCCT TTGCACTGGG TACGCTGCCC AATCTTTTGG 551 CAATCGGCAT TTTTTCCCTG CAACTGAAAA AAATCATGCA AAACCGATAT 601 ATCCGCCTGT GTACAGGATT ATCCGTATCA TTATGGGCAT TATGGAAGCT 651 TGCCGTCCTG TGGCTGTAA This encodes a protein having amino acid sequence <SEQ LD 398>: 1 MNHDITFLTL FLLGFFGGTH CIGMCGGLSS AFALQLPPHI NRFWLILLLN 51 TGRISSYTAI GLMLGLIGQL GISLDQTRVL QNILYTASNL LLLFLGLYLS 101 GISSLAAKIE KIGKPIWRNL NPILNRLLPI KSIPACLAVG ILWGWLPCGL 151 VYSASLYALG SGSATTGGLY MLAFALGTLP NLLAIGIFSL QLKKIMQNRY 201 IRLCTGLSVS LWALWKLAVL WL* In addition, ORF103ng and ORF103-1 show 97.3% identity in 222 aa overlap: 10 20 30 40 50 60 orf103-1.pep MNHDITFLTLFLLGFFGGTHCIGMCGGLSSAFALQLPPHINRFWLILLLNTGRVSSYTAI I I I I I I II II I I II I II I I I I I II I I I II I II I I I I I I I I I I I I II I I I II I I : I I I I I I orf103ng MNHDITFLTLFLLGFFGGTHCIGMCGGLSSAFALQLPPHINRFWLILLLNTGRISSYTAI 10 20 30 40 50 60 70 80 90 100 110 120 orf103-1.pep GLILGLIGQVGVSLDQTRVLQNILYTAANLLLLFLGLYLSGISSLAAKIEKIGKPIWRNL I I : I II II M I: II II I I I I I I I II I I : II I II I II I I I II II II I I I II II I II II I I I orfl03ng GLMLGLIGQLGISLDQTRVLQNILYTASNLLLLFLGLYLSGISSLAAKIEKIGKPIWRNL 70 80 90 100 110 120 130 140 150 160 170 180 orf103-1.pep NPILNRLLPIKSIPACLAVGILWGWLPCGLVYSASLYALGSGSAATGGLYMLAFALGTLP I I I I I II I I I I I I II II I I I I I I II I II I I I II I II II II I II I : I II I II II I I I I I I I orf103ng NPILNRLLPIKSIPACLAVGILWGWLPCGLVYSASLYALGSGSATTGGLYMLAFALGTLP 130 140 150 160 170 180 190 200 210 220 orf103-1.pep NLLAIGIFSLQLKKIMQNRYIRLCTGLSVSLWALWKLAVLWLX II II I II I I I I I I II II I I I I I I I II I I I II I II I II II I I I I orfl03ng NLLAIGIFSLQLKKIMQNRYIRLCTGLSVSLWALWKLAVLWLX 190 200 210 220 Based on this analysis, including the presence of a putative leader sequence (double-underlined) and several putative fransmembrane domains (single-underlined) in the gonococcal protein, it is predicted that the proteins from N. meningitidis and N.gonorrhoeae, and their epitopes, could be useful antigens for vaccines or diagnostics, or for raising antibodies. Example 47 The following partial DNA sequence was identified in N. meningitidis <SEQ ID 399>: 1 ATGGAAAACC AAAGGCCGCT CCTAGGCTTT CGCTTGGCAC TTTTGGCGGC 51 GATGACGTGG GGAACGCTGC CGAT . TCCGT GCGGCAGGTA TTGAAGTTTG 101 TCGATGCGCC GACGCTGGTG TGGGTGCGTT TTACCGTGGC GGCGGCGGTA 151 TTGTTTGTTT TGCTGGCACT GGGCGGGCGG CTGCcGAAGC GGCGaGGATT 201 TTTCTTGGTG CTCATTCAGG CTGCTGCTGC TCGGCGTGGC GGGCATTTCG 251 GCAAACTTTG TGCTGATTGC CCAAGGGCTG CATTATATTT CGCCGACCAC 301 GACGCAGGTT TTGTGGCAGA TTTCGCCGTT TACGATGATT GT GTCGGTG 351 TGTTGGTGTT TAAAGACCGG ATGACTGCCG CTCAGAAAAT CGGCTTGGTT 401 TTGCTGCTTG CCGGTTTGCT TATGTATTTT AACGATAAAT TCGGCGAGTT 451 GTCGGGTTTG GGCGCGTATG C.AAGGGCGT GTTGCTGTGT GCGGCAGGCA 501 GTATGGCATG GGTGTGTAAT GCCGTGGCGC AAAAGCTGCT GTCGGCGCAA 551 TTCGGGCCGC AACAGATTCT GCTGTTGATT TATGCGGCAA GTGCCGCCGT 601 GTTCCTGCCG TTTGCCGAAC CGGCACACAT CGGAAGTATG GACGGTACGT 651 TGGCGTGGGT ATGTATTGCG TATTGCTGCT TGAATACGTT AATCGGTTAC 701 GGCTCGTTCG GCGAGGCGTT GAAACATTGG GAGGCTTCCA AAGTCAGCGC 751 GGTAACAACC TTGCTCCCCG TGTTTACCGT AATAAATACT TTGCTCGGGC 801 ATTATGTGAT GCCTGAAACT TTTGCCGCGC CGGA.. This coπesponds to the .amino acid sequence <SEQ ID 400; ORF104>: 1 MENQRPLLGF RLALLAAMTW GTLPXSVRQV LKFVDAPTLV WVRFTVAAAV 51 LFVLLALGGR LPKRRDFSWC SFRLLLLGVA GISANFVLIA QGLHYISPTT 101 TQVLWQISPF TMI GVLVF KDRMTAAQKI GLVLLLAGLL MYFNDKFGEL 151 SGLGAYXKGV LLCAAGSMAW VCNAVAQKLL SAQFGPQQIL LLIYAASAAV 201 FLPFAEPAHI GSMDGTLAWV CIAYCCLNTL IGYGSFGEAL KHWEASKVSA 251 VTTLLPVFTV INTLLGHYVM PETFAAP... Further work revealed further partial DNA sequence <SEQ ID 401>: 1 ATGGAAAACC AAAGGCCGCT CCTAGGCTTC GCGTTGGCAC TTTTGGCGGC 51 GATGACGTGG GGAACGCTGC CGATTGCCGT GCGGCAGGTA TTGAAGTTTG 101 TCGATGCGCC GACGCTGGTG TGGGTGCGTT TTACCGTGGC GGCGGCGGTA 151 TTGTTTGTTT TGCTGGCACT GGGCGGGCGG CTGCCGAAGC GGCGGGATTT 201 TTCTTGGTGC TCATTCAGGC TGCTGCTGCT CGGCGTGGCG GGCATTTCGG 251 CAAACTTTGT GCTGATTGCC CAAGGGCTGC ATTATATTTC GCCGACCACG 301 ACGCAGGTTT TGTGGCAGAT TTCGCCGTTT ACGATGATTG TTGTCGGTGT 351 GTTGGTGTTT AAAGACCGGA TGACTGCCGC TCAGAAAATC GGCTTGGTTT 401 TGCTGCTTGC CGGTTTGCTT ATGTTTTTTA ACGATAAATT CGGCGAGTTG 451 TCGGGTTTGG GCGCGTATGC GAAGGGCGTG TTGCTGTGTG CGGCAGGCAG 501 TATGGCATGG GTGTGTTATG CCGTGGCGCA AAAGCTGCTG TCGGCGCAAT 551 TCGGGCCGCA ACAGATTCTG CTGTTGATTT ATGCGGCAAG TGCCGCCGTG 601 TTCCTGCCGT TTGCCGAACC GGCACACATC GGAAGTTTGG ACGGTACGTT 651 GGCGTGGGTT TGTTTTGCGT ATTGCTGCTT GAATACGTTA ATCGGTTACG 701 GCTCGTTCGG CGAGGCGTTG AAACATTGGG AGGCTTCCAA AGTCAGCGCG 751 GTAACAACCT TGCTCCCCGT GTTTACCGTA ATA TwwCTT TGCTCGGGCA 801 TTATGTGATG CCTGAAACTT TTGCCGCGCC GGA... This coπesponds to the amino acid sequence <SEQ ID 402; ORF104-1>: 1 MENQRPLLGF ALALLAAMTW GTLPIAVRQV LKFVDAPTLV WVRFTVAAAV 51 LFVLLALGGR LPKRRDFSWC SFRLLLLGVA GISANFVLIA QGLHYISPTT 101 TQVLWQISPF TMIVVGVLVF KDRMTAAQKI GLVLLLAGLL MFFNDKFGEL 151 SGLGAYAKGV LLCAAGSMAW VCYAVAQKLL SAQFGPQQIL LLIYAASAAV 201 FLPFAEPAHI GSLDGTLAWV CFAYCCLNTL IGYGSFGEAL KHWEASKVSA 251 VTTLLPVFTV IXXLLGHYVM PETFAAP... Computer analysis of this amino acid sequence gave the following results: Homology with hypothetical HI0878 protein of H. influenzae (accession number U32769) ORF 104 and HI0878 show 40% aa identity in 277aa overlap: orfl04 4 QRPLLGFRLALLAAMTWGTLPXSVRQVLKFVDAPTLVWXXXXXXXXXXXXXXXXXXXXP- 62 Q+PLLGF AL+ AM WG+LP +++QVL ++A T+VW P HI0878 3 QQPLLGFTFALITAMAWGSLPIALKQVLSVMNAQTIVWYRFIIAAVSLLALLAYKKQLPE 62 orfl04 63 —KRRDFSWCSFRLLLLGVAGISANFVLIAQGLHYISPTTTQVLWQISPFTMIWGVLVF 120 K R ++W ++L+GV G+++NF+L + L+YI P+ Q+ +S F M++ GVL+F HI0878 63 LMKVRQYAW IMLIGVIGLTSNFLLFSSSLNYIEPSVAQIFIHLSSFGMLICGVLIF 118 orfl04 121 KDRMTAAQKIXXXXXXXXXXMYFNDKFGELSGLGAYXKGVLLCAAGSMAWVCNAVAQKLL 180 K+++ QKI ++FND+F +GL Y GV+L G++ WV +AQKL+ HI0878 119 KEKLGLHQKIGLFLLLIGLGLFFNDRFDAFAGLNQYSTGVILGVGGALIWVAYGMAQKLM 178 orfl04 181 SAQFGPQQILLLIYAASAAVFLPFAEPAHIGSMDGTLAWVCIAYCCLNTLIGYGSFGEAL 240 +F QQILL++Y A F+P A+ + + + LA +C YCCLNTLIGYGS+ EAL HI0878 179 LRKFNSQQILLMMYLGCAIAFMPMADFSQVQELT-PLALICFIYCCLNTLIGYGSYAEAL 237 orfl04 241 KHWEASKVSAVTTLLPVFTVINTLLGHYVMPETFAAP 277 W+ SKVS V TL+P+FT++ + + HY P FAAP HI0878 238 NRWDVSKVSWITLVPLFTILFSHIAHYFSPADFAAP 274 Homology with a predicted ORF from N.meninsitidis (strain Al ORF104 shows 95.3% identity over a 277aa overlap with an ORF (ORF104a) from strain A of N. meningitidis: 10 20 30 40 50 60 orf104.pep MEΝQRPLLGFRLALLAAMTWGTLPXSVRQVLKFVDAPTLVWVRFTVAAAVLFVLLALGGR I II I I I I II I I II I I I I I I I I I I : I I I I I I II I I I I I I I I I I I I II M II I I I II I M orf104a MEΝQRPLLGFALALLAAMTWGTLPIAVRQVLKFVDAPTLVWVRFTVAAAVLFVLLALGGR 10 20 30 40 50 60 70 80 90 100 110 120 orfl04.pep LPKRRDFSWCSFRLLLLGVAGISANFVLIAQGLHYISPTTTQVLWQISPFTMIWGVLVF III II II I I II I I I I II II II I I I II I I I I I I I I II I I I I I I I I I I I II I I II I I II I I orf104a LPKWRDFSWCSFRLLLLGVAGISANFVLIAQGLHYISPTTTQVLWQISPFTMIWGVLVF 70 80 90 100 110 120 130 140 150 160 170 180 orf104. pep KDRMTAAQKIGLVLLLAGLLMYFNDKFGELSGLGAYXKGVLLCAAGSMAWVCNAVAQKLL II I I I I I I II I II I I II II IM I II I II I I I I II I I I II I I II II I I II I I I I II I I I orf104a KDRMTAAQKIGLVLLLAGLLMFFNDKFGELSGLGAYAKGVLLCAAGSMAWVCYAVAQKLL 130 140 150 160 170 180 190 200 210 220 230 240 orf104. pep SAQFGPQQILLLIYAASAAVFLPFAEPAHIGSMDGTLAWVCIAYCCLNTLIGYGSFGEAL II I II II II I I I I I I I II II I II I I I I II I I : I II I I I I I : I II I I I I I II I I II I I I I orf104a SAQFGPQQILLLIYAASAAVFLPFAELAHIGSLDGTLAWVCFAYCCLNTLIGYGSFGEAL 190 200 210 220 230 240 250 260 270 orf104. pep KHWEASKVSAVTTLLPVFTVINTLLGHYVMPETFAAP I II I II II II II II I II I I I I : II I I I I I I : I II II orf104a KHWEASKVSAVTTLLPVFTVIFSLLGHYVMPDTFAAPDMNGLGYAGALVWGGAVTAAVG 250 260 270 280 290 300 The complete length ORF 104a nucleotide sequence <SEQ LD 403> is: 1 ATGGAAAACC AAAGGCCGCT CCTAGGCTTC GCGTTGGCAC TTTTGGCGGC 51 GATGACGTGG GGAACGCTGC CGATTGCCGT GCGGCAGGTA TTGAAGTTTG 101 TCGATGCGCC GACGCTGGTG TGGGTGCGTT TTACCGTGGC GGCGGCGGTA 151 TTGTTTGTTT TGCTGGCATT GGGCGGGCGG CTGCCGAAGT GGCGGGATTT 201 TTCTTGGTGC TCATTCAGGC TGCTGCTGCT CGGCGTGGCG GGCATTTCGG 251 CAAACTTTGT GCTGATTGCC CAAGGGCTGC ATTATATTTC GCCGACCACG 301 ACGCAGGTTT TGTGGCAGAT TTCGCCGTTT ACGATGATTG TTGTCGGTGT 351 GTTGGTGTTT AAAGACCGGA TGACTGCCGC TCAGAAAATC GGCTTGGTTT 401 TGCTGCTTGC CGGTTTGCTT ATGTTTTTTA ACGATAAATT CGGCGAGTTG 451 TCGGGTTTGG GCGCGTATGC GAAGGGCGTG TTGCTGTGTG CGGCAGGCAG 501 TATGGCATGG GTGTGTTATG CCGTGGCGCA AAAGCTGCTG TCGGCGCAAT 551 TCGGGCCGCA ACAGATTCTG CTGTTGATTT ATGCGGCAAG TGCCGCCGTG 601 TTCCTGCCGT TTGCCGAACT GGCACACATC GGAAGTTTGG ACGGTACGTT 651 GGCGTGGGTT TGTTTTGCGT ATTGCTGCTT GAATACGTTA ATCGGTTACG 701 GCTCGTTCGG CGAGGCGTTG AAACATTGGG AGGCTTCCAA AGTCAGCGCG 751 GTAACAACCT TGCTCCCCGT GTTTACCGTA ATATTTTCTT TGCTCGGGCA 801 TTATGTGATG CCTGATACTT TTGCCGCGCC GGATATGAAC GGTTTGGGTT 851 ATGCCGGCGC ACTGGTCGTG GTCGGGGGTG CGGTTACGGC GGCGGTGGGG 901 GACAGGCTGT TCAAACGCCG CTAG This encodes a protein having amino acid sequence <SEQ ID 404>: 1 MENQRPLLGF ALALLAAMTW GTLPIAVRQV LKFVDAPTLV WVRFTVAAAV 51 LFVLLALGGR LPKWRDFSWC SFRLLLLGVA GISANFVEIA QGLHYISPTT 101 TQVLWQISPF TMIWGVLVF KDRMTAAQKI GLVLLLAGLL MFFNDKFGEL 151 SGLGAYAKGV LLCAAGSMAW VCYAVAQKLL SAQFGPQQIL LLIYAASAAV 201 FLPFAELAHI GSLDGTLAWV CFAYCCLNTL IGYGSFGEAL KHWEASKVSA 251 VTTLLPVFTV IFSLLGHYVM PDTFAAPDMN GLGYAGAL VGGAVTAAVG 301 DRLFKRR* ORF 104a and ORF 104-1 show 98.2% identity in 277 aa overlap: 10 20 30 40 50 60 orf104a . pep MENQRPLLGFALALLAAMTWGTLPIAVRQVLKFVDAPTLVWVRFTVAAAVLFVLLALGGR I II II I I I I I I II I I I I II I I I I II I II I I II I I I I I II I I I I I I I I I I I II I I II II I I orfl04-l MENQRPLLGFALALLAAMTWGTLPIAVRQVLKFVDAPTLVWVRFTVAAAVLFVLLALGGR 10 20 30 40 50 60 70 80 90 100 110 120 orf104a. pep LPKWRDFSWCSFRLLLLGVAGISANFVLIAQGLHYISPTTTQVLWQISPFTMIWGVLVF III I I I II I I I I II I II I I I II I I I I I I II I II I I II I I I I I I II I II I I I I II I I I II orf104-1 LPKRRDFSWCSFRLLLLGVAGISANFVLIAQGLHYISPTTTQVLWQISPFTMIWGVLVF 70 80 90 100 110 120 130 140 150 160 170 180 orf104a . pep KDRMTAAQKIGLVLLLAGLLMFFNDKFGELSGLGAYAKGVLLCAAGSMAWVCYAVAQKLL I I I I I I I I I I I M M I M I I M M I I I I I I I I I M I I M M I I I I I I I I M M I I I I M I orfl04-l KDRMTAAQKIGLVLLLAGLLMFFNDKFGELSGLGAYAKGVLLCAAGSMAWVCYAVAQKLL 130 140 150 160 170 180 190 200 210 220 230 240 orf104a. pep SAQFGPQQILLLIYAASAAVFLPFAELAHIGSLDGTLAWVCFAYCCLNTLIGYGSFGEAL II I I I II I I I I II I I I I I I I II I I I I II I I I I I II II I I I I I I II II I I II I I I II II I orfl04-l SAQFGPQQILLLIYAASAAVFLPFAEPAHIGSLDGTLAWVCFAYCCLNTLIGYGSFGEAL 190 200 210 220 230 240 250 260 270 280 290 300 orf104a. pep KHWEASKVSAVTTLLPVFTVIFSLLGHYVMPDTFAAPDMNGLGYAGALVWGGAVTAAVG II I II I II II II I I II I I II I I I I I I I II : I I II I orfl04-l KHWEASKVSAVTTLLPVFTVIXXLLGHYVMPETFAAP 250 260 270 Homology with a predicted ORF from N.sonorrhoeae ORF104 shows 93.9% identity over a 277aa overlap with a predicted ORF (ORF104.ng) from N. gonorrhoeae: orf104.pep MENQRPLLGFRLALLAAMTWGTLPXSVRQVLKFVDAPTLVWVRFTVAAAVLFVLLALGGR 60 I II II I II I I I I I I II II I II I I : II I I I II II I II II I I I I I II I I I I II II I I I I I orfl04ng MENQRPLLGFALALLAAMTWGTLPIAVRQVLKFVDAPTLVWVRFTVAAAVLFVLLALGGR 60 orf104.pep LPKRRDFSWCSFRLLLLGVAGISANFVLIAQGLHYISPTTTQVLWQISPFTMIWGVLVF 120 II I I I I I II I I I I II II I : I I II I I I II I II II I I I II II I I I II I I II I I II I I I II I orfl04ng LPKRRDFSWHSFRLLLLGVTGISANFVLIAQGLHYISPTTTQVLWQISPFTMIWGVLVF 120 orf104.pep KDRMTAAQKIGLVLLLAGLLMYFNDKFGELSGLGAYXKGVLLCAAGSMAWVCNAVAQKLL 180 I II I I I I I I I II II I I : I I II : I II I II II I II II I I II I I I I I II I I I I I I II I I I I orf104ng KDRMTAAQKIGLVLLLVGLLMFF DKFGELSGLGAYAKGVLLCAAGSMAWVCYAVAQKLL 180 orf104.pep SAQFGPQQILLLIYAASAAVFLPFAEPAHIGSMDGTLAWVCIAYCCLNTLIGYGSFGEAL 240 II I I I I I I II II II I I I I I II I I I I I II I I : II I II II I :: I I I I I I I II I I I II I I I orf104ng SAQFGPQQILLLIYAASAAVFLLXAEPAHIGSLDGTLAWVCFVYCCLNTLIGYGSFGEAL 240 orf104.pep KHWEASKVSAVTTLLPVFTVINTLLGHYVMPETFAAP 277 I I II I I I I I II I I II II II I I : I II I II I I : II I II orfl04ng KHWEASKVSAVTTLLPVFTVIFSLLGHYVMPDTFAAPDMNGLGYVGALVWGGAVTAAVG 300 The complete length ORF104ng nucleotide sequence <SEQ ID 405> is predicted to encode a protein having amino acid sequence <SEQ ID 406>: 1 MENQRPLLGF ALALLAAMTW GTLPIAVRQV LKFVDAPTLV WVRFTVAAAV 51 LFVLLALGGR LPKRRDFSWH SFRLLLLGVT GISANFVLIA QGLHYISPTT 101 TQVLWQISPF TMIWGVLVF KDRMTAAQKI GLVLLLVGLL MFFNDKFGEL 151 SGLGAYAKGV LLCAAGSMAW VCYAVAQKLL SAQFGPQQIL LLIYAASAAV 201 FLLXAEPAHI GSLDGTLAWV CFVYCCLNTL IGYGSFGEAL KHWEASKVSA 251 VTTLLPVFTV IFSLLGHYVM PDTFAAPDMN GLGYVGALW VGGAVTAAVG 301 DRPFKRR* Further work revealed the complete gonococcal nucleotide sequence <SEQ ID 407>: 1 ATGGAAAACC AAAGGCCGCT CCTAGGCTTC GCGTTGGCAC TTTTGGCGGC 51 GATGACGTGG GGGACGCTGC CGATTGCCGT GCGGCAGGTA TTGAAGTTTG 101 TCGATGCGCC GACGCTGGTG TGGGTGCGTT TTACCGTGGC GGCGGCGGTA 151 TTGTTTGTTT TGCTGGCATT GGGCGGGCGG CTGCCGAAGC GGCGGGATTT 201 TTCTTGGCAT TCATTCAGGC TGCTGCTGCT CGGCGTGACG GGCATTTCGG 251 CAAACTTTGT GCTGATTGCC CAAGGGCTGC ATTATATTTC GCCGACCACG 301 ACGCAGGTTT TGTGGCAGAT TTCGCCGTTT ACGATGATTG TTGTCGGCGT 351 GTTGGTGTTT AAAGACCGGA tgaCTGCCGC GCAGAAAATC GGTTTGGTTT 401 TGCTGCttgT CGGTttgCTT ATGTTTTtta ACGACAAATT CGGCGAGTTG 451 TCGGGTTTGG GCGCGTATGC GAAGGGCGTG TTGCTGTGTG CGGCAGGCAG 501 TATGGCCTGG GTGTGTTATG CCGTGGCGCA AAAGCTGCTG TCGGCGCAAT 551 TCGGGCCGCA ACAGATTCTG CTGTTGATTT ATGCGGcaag tgccgccGTG 601 TTCCtgccgT TTGccgaaCC GGCACACATC GGAAGTTTgg aCGGTACGtt 651 GGCGTGGGTT TGTTTTGTGT ATTGCTGCTT GAATACGTTA ATCGGTTACG 701 GCTCGTTCGG CGAGGCGTTG AAACATTGGG AGGCTTCCAA AGTCAGCGCG 751 GTAACAACCT TGCTCCCCGT GTTTACCGTA ATATTTTCTT TGCTCGGGCA 801 TTATGTGATG CCTGATACTT TTGCCGCGCC GGATATGAAC GGTTTGGGTT 851 ATGTCGGCGC ACTGGTCGTG GTCGGGGGTG CGGTTACGGC GGCGGTGGGG 901 GACAGGCCGT TCAAACGCCG CTAG This coπesponds to the amino acid sequence <SEQ ED 408; ORF104ng-l > • 1 MENQRPLLGF ALALLAAMTW GTLPIAVRQV LKFVDAPTLV WVRFTVAAAV 51 LFVLLALGGR LPKRRDFSWH SFRLLLLGVT GISANFVLIA QGLHYISPTT 101 TQVLWQISPF TMIWGVLVF KDRMTAAQKI GLVLLLVGLL MFFNDKFGEL 151 SGLGAYAKGV LLCAAGSMAW VCYAVAQKLL SAQFGPQQIL LLIYAASAAV 201 FLPFAEPAHI GΞLDGTLAWV CFVYCCLNTL IGYGSFGEAL KHWEASKVSA 251 VTTLLPVFTV IFSLLGHYVM PDTFAAPDMN GLGYVGALW VGGAVTAAVG 301 DRPFKRR* ORF104ng-l and ORF104-1 show 97.5% identity in 277 aa overlap: 10 20 30 40 50 60 orf104-1. pep MENQRPLLGFALALLAAMTWGTLPIAVRQVLKFVDAPTLVWVRFTVAAAVLFVLLALGGR I I II I I I I I I II I I I II I I I II I I I II I I I II I I II I I II I I I II II I I I II I I I I II I I orfl04ng-l MENQRPLLGFALALLAAMTWGTLPIAVRQVLKFVDAPTLVWVRFTVAAAVLFVLLALGGR 10 20 30 40 50 60 70 80 90 100 110 120 orf104-1. pep LPKRRDFSWCSFRLLLLGVAGISANFVLIAQGLHYISPTTTQVLWQISPFTMIWGVLVF II I I II II I I II II I I I I: I I II II I I I II I II I II I I I II II II II II II II II I I II orfl04ng-l LPKRRDFSWHSFRLLLLGVTGISANFVLIAQGLHYISPTTTQVLWQISPFTMIWGVLVF 70 80 90 100 110 120 130 140 150 160 170 180 orf104-1. ep KDRMTAAQKIGLVLLLAGLLMFFNDKFGELSGLGAYAKGVLLCAAGSMAWVCYAVAQKLL I I II I I I I II I I I II I : I I I I I II I I I I I II I I II II II I I I II I I I I I I II I I II I I I I orfl04ng-l KDRMTAAQKIGLVLLLVGLLMFFNDKFGELSGLGAYAKGVLLCAAGSMAWVCYAVAQKLL 130 140 150 160 170 180 190 200 210 220 230 240 orf104-1. pep SAQFGPQQILLLIYAASAAVFLPFAEPAHIGSLDGTLAWVCFAYCCLNTLIGYGSFGEAL I II I II I I II II I II II II I I I II II I I I II I I II II II I I I : I I II I I I II I I I II II I orfl04ng-l SAQFGPQQILLLIYAASAAVFLPFAEPAHIGSLDGTLAWVCFVYCCLNTLIGYGSFGEAL 190 200 210 220 230 240 250 260 270 orfl04-l.pep KHWEASKVSAVTTLLPVFTVIXXLLGHYVMPETFAAP I I II I I II I I I II I II I I I II I I II I II I : II II I orfl04ng-l KHWEASKVSAVTTLLPVFTVIFSLLGHYVMPDTFAAPDMNGLGYVGALVWGGAVTAAVG 250 260 270 280 290 300 In addition, ORF104ng-l shows significant homology with a hypothetical H.influenzae protein: gi 11573895 (U32769) hypothetical [Haemophilus influenzae] Length = 306 Score = 237 bits (598), Expect = 8e-62 Identities = 114/280 (40%), Positives = 168/280 (59%), Gaps = 8/280 (2%) Query: 30 QRPXXXXXXXXXXXMTWGTLPIAVRQVLKFVDAPTLVWXXXXXXXXXXXXXXXXXXXXP- 88 Q+P M WG+LPIA++QVL ++A T+VW P Sbjct: 3 QQPLLGFTFALITAMAWGSLPIALKQVLSVMNAQTIVWYRFIIAAVSLLALLAYKKQLPE 62 Query: 89 —KRRDFSWHSFRLLLLGVTGISANFVLIAQGLHYISPTTTQVLWQISPFTMIWGVLVF 146 K R ++W ++L+GV G+++NF+L + L+YI P+ Q+ +S F M++ GVL+F Sbjct: 63 LMKVRQYAW IMLIGVIGLTSNFLLFSSSLNYIEPSVAQIFIHLSSFGMLICGVLIF 118 Query: 147 KDRMTAAQKIXXXXXXXXXXMFFNDKFGELSGLGAYAKGVLLCAAGSMAWVCYAVAQKLL 206 K+++ QKI +FFND+F +GL Y+ GV+L G++ WV Y +AQKL+ Sbjct: 119 KEKLGLHQKIGLFLLLIGLGLFFNDRFDAFAGLNQYSTGVILGVGGALIWVAYGMAQKLM 178 Query: 207 SAQFGPQQILLLIYAASAAVFLPFAEPAHIGSLDGTLAWVCFVYCCLNTLIGYGSFGEAL 266 +F QQILL++Y A F+P A+ + + L LA +CF+YCCLNTLIGYGS+ EAL Sbjct: 179 LRKFNSQQILLMMYLGCAIAFMPMADFSQVQELT-PLALICFIYCCLNTLIGYGSYAEAL 237 Query: 267 KHWEASKVSAVTTLLPVFTVIFSLLGHYVMPDTFAAPDMN 306 W+ SKVS V TL+P+FT++FS + HY P FAAP++N Sbjct: 238 NRWDVSKVSWITLVPLFTILFSHIAHYFSPADFAAPELN 277 Based on this analysis, including the presence of a putative leader sequence and several putative transmembrane domains in the gonococcal protein, it is predicted that the proteins from N. meningitidis and N.gonorrhoeae, and their epitopes, could be useful antigens for vaccines or diagnostics, or for raising antibodies. Example 48 The following partial DNA sequence was identified in N. meningitidis <SEQ ID 409>: 1 ATGGTAGCTC GTCGGGCTCA TAACCCGAAG GTCGTAGGTT CGAATCCTGT 51 .CCCGCAACC TAATTTCAAA CCCCTCGGTT CAATGCCGAG GG.GTTTTGT 101 T.TTGCCTGT TTCCTGTTTC CTGTTTCCTG CCGCCTCCGT TTTTTGCCGG 151 ATTTTCCTTC CGGCCGCAAT ATCGGAACGG CAGACCGCCG TCTGTTTGCG 201 GTTGCAAATT CAGGCAGTTT GGCTACAATC TTCCGCATTG TCTTCAAGAA 251 AGCCAACCAT GCCGACCGTC CGTTTTACCG AATCCGTCAG CAAACAAGAC 301 CTTGATGCTC TGTTCGAGTG GGCAAAAGCA AGTTACGGTG CAGAAAGTTG 351 CTGGAAAACG CTGTATCTGA ACGGTCysCC TTTGGGCAAC CTGTCGCCGG 401 AATGGGTGGA ACGCGTsmmA AAAGACTGGG AGGCAGGCTG CyCGGAGTCT 451 TCAGACGGCA TTTTTCTGAA TgCGGACGGc TGgCctGATA TGGgCGGAcg 501 cTTACAGCAC CTCGCCCTCG GTTGGCACTG TGCGGGGCTG TTGGACGgsT 551 GGCGCAACGA GTGTTTCGAC CTGACCGACG GCGGCGGCAA CCCCTTGTTC 601 ACGCTCGaAc GCGCCGyTTT mCGTCCTkTC GGACTGCTCA GCCGCGCCGT 651 CCATCTCAAC GGTCTGACCG AATCGGACGG CCGATGGCAT TTCTGGATAG 701 GCAGGCGCAG TCCGCACAAA GCAGTCGATC CCAACAAACT CGACAATACT 751 rCCGCCGGCG GTGTTTCCGG CGGCGAAATG CCGTCTGAAG CCGTGTGTCG 801 CGAAAGCAGC GAAGAAGCCG GTTTGGATAA AACGCTGcTT CCGCTCATCC 851 GCCCGGTATC GCAGCTGCAC AGCCTGCGCT CCGTCAGCCG GGGTGTACAC 901 AATGAAATCC TGTATGTATT CGATGCCGTC CTGCCG ... This coπesponds to the amino acid sequence <SEQ ID 410; ORF105>: 1 MVARRAHNPK VVGSNPXPAT XFQTPRFNAE XVLXLPVSCF LFPAASVFCR 51 IFLPAAISER QTAVCLRLQI QAVWLQSSAL SSRKPTMP2V RFTESVSKQD 101 LDALFEWAKA SYGAESCWKT LYLNGXPLGN LSPEWVERVX KDWEAGCXES 151 SDGIFLNADG WPDMGGRLQH LALGWHCAGL LDGWRNECFD LTDGGGNPLF 201 TLERAXXRPX GLLSRAVHLN GLTESDGRWH FWIGRRSPHK AVDPNKLDNT 251 XAGGVSGGEM PSEAVCRESS EEAGLDKTLL PLIRPVSQLH SLRSVSRGVH 301 NEILYVFDAV LP... Further work revealed the complete nucleotide sequence <SEQ ID 411>: 1 ATGCCGACCG TCCGTTTTAC CGAATCCGTC AGCAAACAAG ACCTTGATGC 51 TCTGTTCGAG TGGGCAAAAG CAAGTTACGG TGCAGAAAGT TGCTGGAAAA 101 CGCTGTATCT GAACGGTCTG CCTTTGGGCA ACCTGTCGCC GGAATGGGTG 151 GAACGCGTCA AAAAAGACTG GGAGGCAGGC TGCTCGGAGT CTTCAGACGG 201 CATTTTTCTG AATGCGGACG GCTGGCCTGA TATGGGCGGA CGCTTACAGC 251 ACCTCGCCCT CGGTTGGCAC TGTGCGGGGC TGTTGGACGG CTGGCGCAAC 301 GAGTGTTTCG ACCTGACCGA CGGCGGCGGC AACCCCTTGT TCACGCTCGA 351 ACGCGCCGCT TTCCGTCCTT TCGGACTGCT CAGCCGCGCC GTCCATCTCA 401 ACGGTCTGAC CGAATCGGAC GGCCGATGGC ATTTCTGGAT AGGCAGGCGC 451 AGTCCGCACA AAGCAGTCGA TCCCAACAAA CTCGACAATA CTGCCGCCGG 501 CGGTGTTTCC GGCGGCGAAA TGCCGTCTGA AGCCGTGTGT CGCGAAAGCA 551 GCGAAGAAGC CGGTTTGGAT AAAACGCTGC TTCCGCTCAT CCGCCCGGTA 601 TCGCAGCTGC ACAGCCTGCG CTCCGTCAGC CGGGGTGTAC ACAATGAAAT 651 CCTGTATGTA TTCGATGCCG TCCTGCCCGA AACCTTCCTG CCTGAAAATC 701 AGGATGGCGA AGTGGCGGGT TTTGAGAAAA TGGACATCGG CGGTCTGTTG 751 GATGCCATGT TGTCGGGAAA CATGATGCAC GACGCGCAAC TGGTTACGCT 801 GGACGCGTTT TGCCGTTACG GTCTGATTGA TGCCGCCCAT CCGCTGTCCG 851 AGTGGCTGGA CGGCATACGT TTATAG This coπesponds to the amino acid sequence <SEQ ID 412; ORF105-1>: 1 MPΓVRFTESV SKQDLDALFE WAKASYGAES CWKTLYLNGL PLGNLSPEWV 51 ERVKKDWEAG CSESSDGIFL NADGWPDMGG RLQHLALGWH CAGLLDGWRN 101 ECFDLTDGGG NPLFTLERAA FRPFGLLSRA VHLNGLTESD GRWHFWIGRR 151 SPHKAVDPNK LDNTAAGGVS GGEMPSEAVC RESSEEAGLD KTLLPLIRPV 201 SQLHSLRSVS RGVHNEILYV FDAVLPETFL PENQDGEVAG FEKMDIGGLL 251 DAMLSGNMMH DAQLVTLDAF CRYGLIDAAH PLSEWLDGIR L* Computer analysis of this amino acid sequence gave the following results: Homology with a predicted ORF from N.meninsitidis (strain Al ORF105 shows 89.4% identity over a 226aa overlap with an ORF (ORF105a) from strain A of N. meningitidis: 60 70 80 90 100 110 orf105. pep ISERQTAVCLRLQIQAVWLQSSALSSRKPTMPTVRFTESVSKQDLDALFEWAKASYGAES I I II I II II II I : I II I II I I I I II I II II orf105a MPTVRFTESVSKHDLDALFEWAKASYGAES 10 20 30 120 130 140 150 160 170 orf105. pep CWKTLYLNGXPLGNLSPEWVERVXKDWEAGCXESSDGIFLNADGWPDMGGRLQHLALGWH I II I I II II II I II I I I I: II I I I II I II II II I I I II II I II I II I II II I I : orf105a CWKTLYLNGLPLGNLSPEWAERVKKDWEAGCSESSDGIFLNADGWPDMGRRLQHLARIWK 40 50 60 70 80 90 180 190 200 210 220 230 orf105. pep CAGLLDGWRNECFDLTDGGGNPLFTLERAXXRPXGLLSRAVHLNGLTESDGRWHFWIGRR II I I I II : II I II I I II : II I I : II II II I II II II I I I II : I I I II I II I I I II orf105a EAGLLHGWRDECFDLTDGGSNPLFALERAAFRPFGLLSRAVHLNGLVESDGRWHFWIGRR 100 110 120 130 140 150 240 250 260 270 280 290 orf105.pep SPHKAVDPNKLDNTXAGGVSGGEMPSEAVCRESSEEAGLDKTLLPLIRPVSQLHΞLRSVS I II II I I I : I I II I I II II : II : I I I : II I II II I II I I I I II I I I II I I I II I II II orf105a SPHKAVDPDKLDNTAAGGVSSGELPSETVCRESSEEAGLDKTLLPLIRPVSQLHSLRPVS 160 170 180 190 200 210 300 310 orf105. pep RGVHNEILYVFDAVLP I II II I I I II I I I I II orf105a RGVHNEILYVFDAVLPETFLPENQDGEVAGFEKMDIGGLLAAMLSGNMMHDAQLVTLDAF 220 230 240 250 260 270 The complete length ORF 105 a nucleotide sequence <SEQ ID 413> is: 1 ATGCCGACCG TCCGTTTTAC CGAATCCGTC AGCAAACACG ACCTTGATGC 51 CCTATTCGAG TGGGCAAAGG CAAGTTACGG TGCGGAAAGT TGCTGGAAAA 101 CGCTGTATCT GAACGGTCTG CCTTTGGGCA ATCTGTCGCC GGAATGGGCG 151 GAGCGCGTCA AAAAAGACTG GGAGGCAGGC TGCTCGGAGT CTTCAGACGG 201 CATTTTCCTG AATGCGGACG GCTGGCCAGA TATGGGCAGA CGCTTGCAGC 251 ACCTCGCCCG AATATGGAAA GAAGCGGGAC TGCTTCACGG CTGGCGCGAC 301 GAGTGTTTCG ACCTGACCGA CGGCGGCAGC AATCCCTTGT TCGCGCTCGA 351 ACGCGCCGCT TTCCGTCCGT TCGGACTGCT CAGCCGCGCC GTCCATCTCA 401 ACGGTTTGGT CGAATCGGAC GGCCGATGGC ATTTCTGGAT AGGCAGGCGC 451 AGTCCGCACA AAGCAGTCGA TCCCGACAAA CTCGACAATA CTGCCGCCGG 501 CGGTGTTTCC AGCGGTGAAT TGCCGTCTGA AACCGTGTGT CGCGAAAGCA 551 GCGAAGAAGC CGGTTTGGAT AAAACGCTGC TTCCGCTCAT CCGCCCGGTA 601 TCGCAGCTGC ACAGCCTGCG CCCCGTCAGC CGGGGTGTGC ACAATGAAAT 651 CCTGTATGTA TTCGATGCCG TCCTGCCCGA AACCTTCCTG CCTGAAAATC 701 AGGATGGCGA AGTGGCGGGT TTTGAGAAAA TGGACATCGG CGGTCTGTTG 751 GCTGCCATGT TGTCGGGAAA CATGATGCAC GACGCGCAAC TGGTTACGCT 801 GGACGCGTTT TGCCGTTACG GTCTGATTGA TGCCGCCCAT CCGCTGTCCG 851 AGTGGCTGGA CGGCATACGT TTATAG This encodes a protein having amino acid sequence <SEQ ID 414>: 1 MPTVRFTESV SKHDLDALFE WAKASYGAES CWKTLYLNGL PLGNLSPEWA 51 ERVKKDWEAG CSESSDGIFL NADGWPDMGR RLQHLARIWK EAGLLHGWRD 101 ECFDLTDGGS NPLFALERAA FRPFGLLSRA VHLNGLVESD GRWHFWIGRR 151 SPHKAVDPDK LDNTAAGGVS SGELPSETVC RESSEEAGLD KTLLPLIRPV 201 SQLHSLRPVS RGVHNEILYV FDAVLPETFL PENQDGEVAG FEKMDIGGLL 251 AAMLSGNMMH DAQLVTLDAF CRYGLIDAAH PLSEWLDGIR L* ORF105a and ORF105-1 show 93.8% identity in 291 aa overlap: 10 20 30 40 50 60 orf105a . pep MPTVRFTESVSKHDLDALFEWAKASYGAESCWKTLYLNGLPLGNLSPEWAERVKKDWEAG I II II I II I I I I : II II I I II I II I II I I I I I I I I I I I I I I I I I I II I I : II I I I I II I I orf105-1 MPTVRFTESVSKQDLDALFEWAKASYGAESCWKTLYLNGLPLGNLSPEWVERVKKDWEAG 10 20 30 40 50 60 70 80 90 100 110 120 orf105a. pep CSESSDGIFLNADGWPDMGRRLQHLARIWKEAGLLHGWRDECFDLTDGGSNPLFALERAA I II I I I II I I II II I I I II I I I II I I : I II I I I I : II II I I I I I : I I II : II I II orf105-1 CSESSDGIFLNADGWPDMGGRLQHLALGWHCAGLLDGWRNECFDLTDGGGNPLFTLERAA 70 80 90 100 110 120 130 140 150 160 170 180 orf105a . pep FRPFGLLSRAVHLNGLVESDGRWHFWIGRRSPHKAVDPDKLDNTAAGGVSSGELPSETVC I I I I I I I I I I I I I I I I: I II I I I I I I I II I I I II I I I I : I I II I I II I I I: I I: I I I: I I orf105-1 FRPFGLLSRAVHLNGLTESDGRWHFWIGRRSPHKAVDPNKLDNTAAGGVSGGEMPSEAVC 130 140 150 160 170 180 190 200 210 220 230 240 orf105a . pep RESSEEAGLDKTLLPLIRPVSQLHSLRPVSRGVHNEILYVFDAVLPETFLPENQDGEVAG I I II II I I I I I I I I II II I II I II I II I I II I I II I I II I I II II I I I II II II II I I I orf105-1 RESSEEAGLDKTLLPLIRPVSQLHSLRSVSRGVHNEILYVFDAVLPETFLPENQDGEVAG 190 200 210 220 230 240 250 260 270 280 290 orf105a. pep FEKMDIGGLLAAMLSGNMMHDAQLVTLDAFCRYGLIDAAHPLSEWLDGIRLX II I I I II II I II I I II II I I II I I I I I I I I II I I I I II I I II I I I I I I I II orf105-1 FEKMDIGGLLDAMLSGNMMHDAQLVTLDAFCRYGLIDAAHPLSEWLDGIRLX 250 260 270 280 290 Homology with a predicted ORF from N.sonorrhoeae ORF105 shows 87.5% identity over a 312aa overlap with a predicted ORF (ORF105.ng) from N gonorrhoeae: orf105.pep MVARRAHNPKWGSNPXPATXFQTPRFNAEXVLXLPVSCFLFPAASVFCRIFLPAAISER 60 I II II I I I II I I I II I III : I I I I II II II II II I I I I II I I II II I II I I orf105ng MVARRAHNPKWGSNPAPATKYQTPRFNAEGVLF FLFPAASVFCRIFLPAAISER 55 orf105.pep QTAVCLRLQIQAVWLQSSALSSRKPTMPTVRFTESVSKQDLDALFEWAKASYGAESCWKT 120 I M II I I I II I I I II I I I I I I I IM II I I I I I II I I I I I I II II I II I I I I II I II I I orf105ng QAAVCLRLQIQAVWLQSSALCSRKPAMPTVRFTESVSKQDLDALFERAKASYGAESCWKT 115 orf105.pep LYLNGXPLGNLSPEWVERVXKDWEAGCXESSDGIFLNADGWPDMGGRLQHLALGWHCAGL 180 I I II I II I II II I : IM I II I II I II I : I I I I I II I I I II I II I I I I I M III orf105ng LYLNRLPLGNLSPEWAERIKKDWEAGCSESSNGIFLNADGWPDMGGRLQHLARTWNKAGL 175 orf105.pep LDGWRNECFDLTDGGGNPLFTLERAXXRPXGLLSRAVHLNGLTESDGRWHFWIGRRSPHK 240 I I II I I I I II I I I I I I I II II II I II III I I II I I I I: I IM I II I I II I I I I I I orfl05ng LHGWRNECFDLTDGGGNPLFTLERAAFRPFGLLIRAVHLNGLVESNGRWHFWIGRRSPHK 235 orf105. pep AVDPNKLDNTXAGGVSGGEMPSEAVCRESSEEAGLDKTLLPLIRPVSQLHSLRSVSRGVH 300 I II I : I I I I : I I I I I I II II I I I I I II I II II I I II I : I I II II I I I I I I I I II I orfl05ng AVDPGKLDNIAGGGVSGGEMPSEAVCRESSEEAGLDKTLFPLIRPVSRLHSLRPVSRGVH 295 orf105. pep NEILYVFDAVLP 312 I II I II I II II I orfl05ng NEILYVFDAVLPETFLPENQDGEVAGFEKMDIGGLLDAMLΞKNMMHDAQLVTLDAFYRYG 355 A complete length ORF105ng nucleotide sequence <SEQ LD 415> was predicted to encode a protein having amino acid sequence <SEQ ID 416>: 1 MVARRAHNPK WGSNPAPAT KYQTPRFNAE GVLFFLFPAA SVFCRIFLPA 51 AISERQAAVC LRLQIQAVWL QSSALCSRKP AMPTVRFTES VSKQDLDALF 101 ERAKASYGAE SCWKTLYLNR LPLGNLSPEW AERIKKDWEA GCSESSNGIF 151 LNADGWPDMG GRLQHLARTW NKAGLLHGWR NECFDLTDGG GNPLFTLERA 201 AFRPFGLLIR AVHLNGLVES NGRWHFWIGR RSPHKAVDPG KLDNIAGGGV 251 SGGEMPSEAV CRESSEEAGL DKTLFPLIRP VSRLHSLRPV SRGVHNEILY 301 VFDAVLPETF LPENQDGEVA GFEKMDIGGL LDAMLSKNMM HDAQLVTLDA 351 FYRYGLIDAA HPLSEWLDGI RL* Further work revealed the complete nucleotide sequence <SEQ ID 417>: 1 ATGCCGACCG TCCGTTTTAC CGAATCCGTC AGCAAACAAG ACCTTGATGC 51 CCTGTTCGAG CGGGCAAAAG CAAGTTACGG TGCCGAAAGT TGCTGGAAAA 101 CGCTGTATCT GAACCGTCTT CCTTTGGGCA ATCTGTCGCC GGAATGGGCT 151 GAGCGCATCA AAAAAGACTG GGAGGCAGGC TGCTCCGAGT CTTCAGACGG 201 CATTTTTCTG AATGCGGACG GCTGGCCGGA TATGGGCGGA CGCTTGCAGC 251 ACCTCGCCCG CACATGGAAC AAGGCGGGGC TGCTTCACGG ATGGCGCAAC 301 GAGTGTTTCG ACCTGACCGA CGGCGGCGGC AACCCCTTGT TCACGCTCGA 351 ACGCGCCGCT TTCCGTCCGT TCGGACTACT CAGCCGCGCC GTCCATCTCA 401 ACGGTTTGGT CGAATCGAAC GGCAGATGGC ATTTTTGGAT AGGCAGGCGC 451 AGTCCGCACA AAGCAGTCGa tcCCGGCAAG CTCGACAATA TTGCCGGCGG 501 CGGTGTTTCC GGCGGCGAAA TGCCGTCTGA AGCCGTGTGC CGCGAAAGCA 551 GCGAAGAAGC CGGTTTGGAT AAAACGCTGT TTCCGCTCAT CCGCCCAGTA 601 TCGCGGCTGC ACAGCCTTCG CCCCGTCAGC CGAGGTGTGC ACAATGAAAT 651 CCTGTATGTG TTCGATGCCG TCCTGCCCGA AACCTTCCTG CCTGAAAATC 701 AGGATGGCGA GGTAGCGGGT TTTGAAAAGA TGGACATTGG CGGCCTATTG 751 GATGCCATGT TGTCGAAAAA CATGATGCAC GACGCGCAAC TGGTTACGCT 801 GGACGCGTTT TACCGTTACG GTCTGATTGA TGCCGCCCAT CCGCTGTCCG 851 AGTGGCTGGA CGGCATACGT TTATAG This coπesponds to the amino acid sequence <SEQ ID 418; ORF105ng-l>: 1 MPTVRFTESV SKQDLDALFE RAKASYGAES CWKTLYLNRL PLGNLSPEWA 51 ERIKKDWEAG CSESSDGIFL NADGWPDMGG RLQHLARTWN KAGLLHGWRN 101 ECFDLTDGGG NPLFTLERAA FRPFGLLSRA VHLNGLVESN GRWHFWIGRR 151 SPHKAVDPGK LDNIAGGGVS GGEMPSEAVC RESSEEAGLD KTLFPLIRPV 201 SRLHSLRPVS RGVHNEI YV FDAVLPETFL PENQDGEVAG FEKMDIGGLL 251 DAMLSKNMMH DAQLVTLDAF YRYGLIDAAH PLSEWLDGIR L* ORG105ng-l and ORF105-1 show 93.5% identity in 291 aa overlap: 10 20 30 40 50 60 orf105-1.pep MPTVRFTESVSKQDLDALFEWAKASYGAESCWKTLYLNGLPLGNLSPEWVERVKKDWEAG I II I II I I I II II I I I II I I I I II I I II II I II II I I II I I I I I I I I : II: II II II I orfl05ng-l MPTVRFTESVSKQDLDALFERAKASYGAESCWKTLYLNRLPLGNLSPEWAERIKKDWEAG 10 20 30 40 50 60 70 80 90 100 110 120 orf105-1. pep CSESSDGIFLNADGWPDMGGRLQHLALGWHCAGLLDGWRNECFDLTDGGGNPLFTLERAA II I I I I II II I I I I I I II I I I I I II I I: I II I I I I I I I I II I I I I I II I I I II II I orfl05ng-l CSESSDGIFLNADGWPDMGGRLQHLARTWNKAGLLHGWRNECFDLTDGGGNPLFTLERAA 70 80 90 100 110 120 130 140 150 160 170 180 orf105-1. pep FRPFGLLSRAVHLNGLTESDGRWHFWIGRRSPHKAVDPNKLDNTAAGGVSGGEMPSEAVC I I II II I II I I I I I I I : I I M I I I I I I I I I I II I II I I : II I I I : I II I II I I II II II orfl05ng-l FRPFGLLSRAVHLNGLVESNGRWHFWIGRRSPHKAVDPGKLDNIAGGGVSGGEMPSEAVC 130 140 150 160 170 180 190 200 210 220 230 240 orf105-1. pep RESSEEAGLDKTLLPLIRPVSQLHSLRSVSRGVHNEILYVFDAVLPETFLPENQDGEVAG II I I I I II I II I I : I II I II I: I I I I I I I I II I I I I I I II I I I I I I II I II I I II II II orfl05ng-l RESSEEAGLDKTLFPLIRPVSRLHSLRPVSRGVHNEILYVFDAVLPETFLPENQDGEVAG 190 200 210 220 230 240 250 260 270 280 290 orf105-1.pep FEKMDIGGLLDAMLSGNMMHDAQLVTLDAFCRYGLIDAAHPLSEWLDGIRLX I I I I I I I I I I II I II II I I I II I II II I I I I I I I II I I II I I I I I I II I I orfl05ng-l FEKMDIGGLLDAMLSKNMMHDAQLVTLDAFYRYGLIDAAHPLSEWLDGIRLX 250 260 270 280 290 Furthermore, ORF105ng-l shows homology with a yeast enzyme: sp|P41888 |TNR3_SCHPO THIAMIN PYROPHOSPHOKINASE (TPK) (THIAMIN KINASE) >gi 11076928 Ipir M S52350 thiamin pyrophosphokinase (EC 2.7.6.2) - fission yeast (Schizosaccharomyces pombe) >gi 1666111 (X84417) thiamin pyrophosphokinase [Schizosaccharomyces pombe] >gi | 2330852 | gnl | PID | e334056 (Z98533) thiamin pyrophosphokinase [Schizosaccharomyces pombe] Length = 569 Score = 105 bits (259) , Expect = 4e-22 Identities = 64/192 (33%), Positives = 94/192 (48%), Gaps = 3/192 (1%) Query: 268 NKAGLLHGWRNECFDLTDGGGNPLFTLERAAFRPFGLLSRAVHLNGLVESNGRW—HFWI 441 N G+ WRNE + + P+ +ER F FG LS VH + + W+ Sbjct: 96 NTFGIADQWRNELYTVYGKSKKPVLAVERGGFWLFGFLSTGVHCTMYIPATKEHPLRIWV 155 Query: 442 GRRSPHKAVDPGKLDNIAGGGVSGGEMPSEAVCRESSEEAGLDKTLFPLIRPVΞRLHSLR 621 RRSP K P LDN GG++ G+ + +E SEEA LD + LI P + ++ Sbjct: 156 PRRSPTKQTWPNYLDNSVAGGIAHGDSVIGTMIKEFSEEANLDVSSMNLI-PCGTVSYIK 214 Query: 622 PVSRG-VHNEILYVFDAVLPETFLPENQDGEVAGFEKMDIGGLLDAMLSKNMMHDAQLVT 798 R + E+ YVFD + + +P DGEVAGF + + +L + K+ + LV Sbjct: 215 MEKRHWIQPELQYVFDLPVDDLVIPRINDGEVAGFSLLPLNQVLHELELKSFKPNCALVL 274 Query: 799 LDAFYRYGLIDAAHP 843 LD R+G+I HP Sbjct: 275 LDFLIRHGIITPQHP 289 Based on this analysis, including the presence of a putative transmembrane domain in the gonococcal protein, it is predicted that the proteins from N. meningitidis and N.gonorrhoeae, and their epitopes, could be useful antigens for vaccines or diagnostics, or for raising antibodies. Example 49 The following DNA sequence, believed to be complete, was identified in N. meningitidis <SEQ ED 419>: 1 ATGAATAGAC CCAAGCAACC CTTCTTCCGT CCCGAAGTCG CCGTTGCCCG 51 CCAAACCAGC CTGACGGGTA AAGTGATTCT GACACGACCG TTGTCATTTT 101 CCCTATGGAC GACATTTGCA TCGATATCTG CGTTATTGAT TATCCTGTTT 151 TTGATATTTG GTAACTATAC GCGAAAGACA ACAGTGGAGG GACAAATTTT 201 ACCTGCATCG GGCGTAATCA GGGTGTATGC ACCGgATACG rGkACAATTA 251 CAGCGAAATT CGTGGAAGAT GGmsAAAAGG TTAAGGCTGG CGACAAGCTA 301 TTTGCGCTTT CGACCTCACG TTTCGGCGCA GGAGGTAGCG TGCAGCAGCA 351 GTTGAAAACG GAGGCAGTTT TGAAGAAAAC GTTGGCAGAA CAGGAACTGG 401 GTCGTCTGAA GCTGATACAC GGGAATGAAA CGCGCAgCcT TAAAGCAACT 451 GTCGAACGTT TGGAAAACCA GGAACTCCAT ATTTCGCAAC AGATAGACGG 501 TCAGAAAAGG CGCATTAGAC TTGCGGAAGA AATGTTGCAG AAATATCGTT 551 TCCTATCCGC . CAATGA This coπesponds to the amino acid sequence <SEQ ID 420; ORF107>: 1 MNRPKQPFFR PEVAVARQTS LTGKVILTRP LSFSLWTTFA SISALLIILF 51 LIFGNYTRKT TVEGQILPAS GVIRVYAPDT XTITAKFVED GXKVKAGDKL 101 FALSTSRFGA GGSVQQQLKT EAVLKKTLAE QELGRLKLIH GNETRSLKAT 151 VERLENQELH ISQQIDGQKR RIRLAEEMLQ KYRFLSXQ* Computer analysis of this amino acid sequence gave the following results: Homology with a predicted ORF from N.meninsitidis (sfrain Al ORF107 shows 97.8% identity over a 186aa overlap with an ORF (ORF107a) from strain A of N. meningitidis: 10 20 30 40 50 60 orfl07.pep MNRPKQPFFRPEVAVARQTSLTGKVILTRPLSFSLWTTFASISALLIILFLIFGNYTRKT I I II I II II I II I I I I II II II I II II I I I II II I I I I I II I I I II II I I II I I II I I II orfl07a MNRPKQPFFRPEVAVARQTSLTGKVILTRPLSFSLWTTFASISALLIILFLIFGNYTRKT 10 20 30 40 50 60 70 80 90 100 110 120 orf107.pep TVEGQILPASGVIRVYAPDTXTITAKFVEDGXKVKAGDKLFALSTSRFGAGGSVQQQLKT II II I I I I II II I I I II I I I II I II I III I I II II I I II I I I I I I I I I I II I I I II orfl07a TVEGQILPASGVIRVYAPDTGTITAKFXEDGEKVKAGDKLFALSTSRFGAGDSVQQQLKT 70 80 90 100 110 120 130 140 150 160 170 180 orf107.pep EAVLKKTLAEQELGRLKLIHGNETRSLKATVERLENQELHISQQIDGQKRRIRLAEEMLQ I II I I II I I I II I I I II I I II II I II I I I I II II I II II I I I II II II I II I II I I I I I I orfl07a EAVLKKTLAEQELGRLKLIHGNETRSLKATVERLENQELHISQQIDGQKRRIRLAEEMLQ 130 140 150 160 170 180 189 orf107.pep KYRFLSXQX I II I I I orfl07a KYRFLSANDAVPKQEMMNVKAELLEQKAKLDAYRREEVGLLQEIRTQNLTLXSLPQAAX 190 200 210 220 230 The complete length ORF 107a nucleotide sequence <SEQ ID 421 > is: 1 ATGAATAGAC CCAAGCAACC NTTCTTCCGT CCCGAAGTCG CCGTTGCCCG 51 CCAAACCAGC CTGACGGGTA AAGTGATTCT GACACGACCG TTGTCATTTT 101 CCCTATGGAC GACATTTGCA TCGATATCTG CGTTATTGAT TATCCTGTTT 151 TTGATATTTG GTAACTATAC GCGAAAGACA ACAGTGGAGG GACAAATTTT 201 ACCTGCATCG GGCGTAATCA GGGTGTATGC ACCGGATACG GGGACAATTA 251 CNGCGAAATT CNTGGAAGAT GGAGAAAAGG TTAAGGCTGG CGACAAGCTA 301 TTTGCGCTTT CGACCTCACG TTTCGGCGCA GGAGATAGCG TGCAGCAGCA 351 GTTGAAAACG GAGGCAGTTT TGAAGAAAAC GTTGGCAGAA CAGGAACTGG 401 GTCGTCTGAA GCTGATACAC GGGAATGAAA CGCGCAGCCT TAAAGCAACT 451 GTCGAACGTT TGGAAAACCA GGAACTCCAT ATTTCGCAAC AGATAGACGG 501 TCAGAAAAGG CGCATTAGAC TTGCGGAAGA AATGTTGCAG AAATATCGTT 551 TCCTATCCGC CAATGATGCA GTGCCAAAAC AAGAAATGAT GAATGTCAAG 601 GCAGAGCTTT TAGAGCAGAA AGCCAAACTT GATGCCTACC GCCGAGAAGA 651 AGTCGGGCTG CTTCAGGAAA TCCGCACGCA GAATCTGACA TTGGNNAGCC 701 TCCCCCAAGC GGCATGA This encodes a protein having amino acid sequence <SEQ ED 422>: 1 MNRPKQPFFR PEVAVARQTS LTGKVILTRP LSFSLWTTFA SISALLIILF 51 LIFGNYTRKT TVEGQILPAS GVIRVYAPDT GTITAKFXED GEKVKAGDKL 101 FALSTSRFGA GDSVQQQLKT EAVLKKTLAE QELGRLKLIH GNETRSLKAT 151 VERLENQELH ISQQIDGQKR RIRLAEEMLQ KYRFLSANDA VPKQEMMNVK 201 AELLEQKAKL DAYRREEVGL LQEIRTQNLT LXSLPQAA* Homology with a predicted ORF from N.sonorrhoeae ORF107 shows 95.7% identity over a 188aa overlap with a predicted ORF (ORF107.ng) from N. gonorrhoeae: orf107.pep MNRPKQPFFRPEVAVARQTSLTGKVILTRPLSFSLWTTFASISALLIILFLIFGNYTRKT 60 I II I I I I I II I I I I : II I II II I I II I I I I II II I I I I II I I I II II I I I II II I I I I I I orfl07ng MNRPKQPFFRPEVAIARQTSLTGKVILTRPLSFSLWTTFASISALLIILFLIFGNYTRKT 60 orf107.pep TVEGQILPASGVIRVYAPDTXTITAKFVEDGXKVKAGDKLFALSTSRFGAGGSVQQQLKT 120 I : I I II II I I II II I I I I I I I I I I I II II I I I II I I I I I I II II II II I II II I II I I orfl07ng TMEGQILPASGVIRVYAPDTGTITAKFVEDGEKVKAGDKLFALSTSRFGAGGSVQQQLKT 120 orf107.pep EAVLKKTLAEQELGRLKLIHGNETRSLKATVERLENQELHISQQIDGQKRRIRLAEEMLQ 180 II I I I II I I I I I I II II I I I II I II I I I I I I I II I I : I I II I I I I I I I I I I I II I I II : orfl07ng EAVLKKTLAEQELGRLKLIHENETRSLKATVERLENQKLHISQQIDGQKRRIRLAEEMLR 180 orf107.pep KYRFLSXQ 188 II I I I I I orfl07ng KYRFLSAQ 1 The complete length ORF107ng nucleotide sequence <SEQ ID 423> is predicted to encode a protein having amino acid sequence <SEQ ED 424>: 1 MNRPKQPFFR PEVAIARQTS LTGKVILTRP LSFSLWTTFA SISALLIILF 51 LIFGNYTRKT TMEGQILPAS GVIRVYAPDT GTITAKFVED GEKVKAGDKL 101 FALSTSRFGA GGSVQQQLKT EAVLKKTLAE QELGRLKLIH ENETRSLKAT 151 VERLENQKLH ISQQIDGQKR RIRLAEEMLR KYRFLSAQ* Based on the presence of a putative ransmembrane domain in the gonococcal protein, it is predicted that the proteins from N. meningitidis .and N.gonorrhoeae, and their epitopes, could be useful antigens for vaccines or diagnostics, or for raising antibodies. Example 50 The following DNA sequence, believed to be complete, was identified in N. meningitidis <SEQ LD 425>: 1 ATGCTGAATA CTTTTTTTGC CGTATTGGGC GGCTGCCTGC TGCT . TTGCC 51 GTGCGGCAAA TCCGTAAATA CGGCGGTACA GCCGCAAAAC GCGGTACAAA 101 GCGCGCCGAA ACCGGTTTTC AAAGTCATAT ATATCGACAA TACGGCGATT 151 GCCGGTTTGG ATTTGGGACA AAGCAGCGAA GGCAAAACCA ACGACGGCAA 201 AAAACAAATC AGTTATCCGA TTAAAGGCTT GCCGGAACAA AATGTTATCC 251 GACTGATCGG CAAGCATCCC GGCGACTTGG AAGCCGTCAG CGGCAAATGT 301 ATGGAAACCG ATGATAAGGA CAGTCCGGCA GGTTGGGCAG AAAACGGCGT 351 GTGCCATACC TTGTTTGCCA AACTGGTGGG CAATATCGCC GAAGACGGCG 401 GCAAACTGAC GGATTACCTA GTTTCGCATG CCGCCCTGCA ACCCTATCAG 451 GCAGGCAAAA GCGGCTATGC CGCCGTGCAG AACGGACGCT ATGTGCTGGA 501 AATCGACAGC GAAGGGGCGT TTTATTTCCG CCGCCGCCAT TATTGA This coπesponds to the amino acid sequence <SEQ ID 426; ORF108>: 1 MLNTFFAVLG GCLLXLPCGK SVNTAVQPQN AVQSAPKPVF KVIYIDNTAI 51 AGLDLGQSSE GKTNDGKKQI SYPIKGLPEQ NVIRLIGKHP GDLEAVSGKC 101 METDDKDSPA GWAENGVCHT LFAKLVGNIA EDGGKLTDYL VSHAALQPYQ 151 AGKSGYAAVQ NGRYVLEIDS EGAFYFRRRH Y* Further work revealed the following DNA sequence <SEQ LD 427>: 1 ATGCTGAAAA CATCTTTTGC CGTATTGGGC GGCTGCCTGC TGCTTGCCGC 51 CTGCGGCAAA TCCGAAAATA CGGCGGAACA GCCGCAAAAC GCGGTACAAA 101 GCGCGCCGAA ACCGGTTTTC AAAGTCAAAT ATATCGACAA TACGGCGATT 151 GCCGGTTTGG ATTTGGGACA AAGCAGCGAA GGCAAAACCA ACGACGGCAA 201 AAAACAAATC AGTTATCCGA TTAAAGGCTT GCCGGAACAA AATGTTATCC 251 GACTGATCGG CAAGCATCCC GGCGACTTGG AAGCCGTCAG CGGCAAATGT 301 ATGGAAACCG ATGATAAGGA CAGTCCGGCA GGTTGGGCAG AAAACGGCGT 351 GTGCCATACC TTGTTTGCCA AACTGGTGGG CAATATCGCC GAAGACGGCG 401 GCAAACTGAC GGATTACCTA GTTTCGCATG CCGCCCTGCA ACCCTATCAG 451 GCAGGCAAAA GCGGCTATGC CGCCGTGCAG AACGGACGCT ATGTGCTGGA 501 AATCGACAGC GAAGGGGCGT TTTATTTCCG CCGCCGCCAT TATTGA This coπesponds to the amino acid sequence <SEQ ID 428; ORF108-1>: 1 MLKTSFAVLG GCLLLAACGK SENTAEQPQN AVQSAPKPVF KVKYIDNTAI 51 AGLDLGQSSE GKTNDGKKQI SYPIKGLPEQ NVIRLIGKHP GDLEAVSGKC 101 METDDKDSPA GWAENGVCHT LFAKLVGNIA EDGGKLTDYL VSHAALQPYQ 151 AGKSGYAAVQ NGRYVLEIDS EGAFYFRRRH Y* Computer analysis of this amino acid sequence gave the following results: Homology with a predicted ORF from N.sonorrhoeae ORF108 shows 88.4% identity over a 181aa overlap with a predicted ORF (ORF108.ng) from N gonorrhoeae: orf108. pep MLΝTFFAVLGGCLLXLPCGKSVΝTAVQPQΝAVQSAPKPVFKVIYIDΝTAIAGLDLGQSSE 60 II: I I I II I I II II I I I I I I II I I : II I II I II II I I I I II I I I I II I I I I orfl08ng MLKIPFAVLGGCLLLAACGKSEΝTAEQPQΝAAQSAPKPVFKVKYIDΝTAIAGLALGQSSE 60 orf108.pep GKTΝDGKKQISYPIKGLPEQΝVIRLIGKHPGDLEAVSGKCMETDDKDSPAGWAEΝGVCHT 120 I II I II I I II I I II I I I II IMM I I I II: I I II I I I II II I I I : I : I II I II II I I orfl08ng GKTΝDGKKQISYPIKGLPEQΝAVRLTGKHPΝDLEAWGKCMETDGKDAPSGWAEΝGVCHT 120 orf108. pep LFAKLVGΝIAEDGGKLTDYLVSHAALQPYQAGKSGYAAVQΝGRYVLEIDSEGAFYFRRRHY 181 I II I I I I I I I I I I I II I I I IM M I I I I II I II I I I I I I II I I II II I I II I II I I I I II I orfl08ng LFAKLVGΝIAEDGGKLTDYLISHSALQPYQAGKSGYAAVQΝGRYVLEIDSEGAFYFRRRHY 181 ORF108-1 shows 92.3% identity with ORF108ng over the same 181 aa overlap: orf108-1.pep MLKTSFAVLGGCLLLAACGKSEΝTAEQPQΝAVQSAPKPVFKVKYIDΝTAIAGLDLGQSSE 60 III I I II II I I I II II II I II I I II I I I I : I II I II II II I I I I I I I II I I II II I I orfl08ng-l MLKIPFAVLGGCLLLAACGKSEΝTAEQPQΝAAQSAPKPVFKVKYIDΝTAIAGLALGQSΞE 60 orf108-1.pep GKTNDGKKQISYPIKGLPEQΝVIRLIGKHPGDLEAVSGKCMETDDKDSPAGWAEΝGVCHT 120 II I I II I I II I II II II I I II :: II II I I : II I II II I I II I I I : I : II I I I I II II orfl08ng-l GKTNDGKKQISYPIKGLPEQNAVRLTGKHPNDLEAWGKCMETDGKDAPSGWAENGVCHT 120 orf108-1. pep LFAKLVGNIAEDGGKLTDYLVSHAALQPYQAGKSGYAAVQNGRYVLEIDSEGAFYFRRRHY 181 I I I II I I I I I I I I I I I I I I IM I: I I I II I II I II II I II II I I I II I I II I II I II I I I I orfl08ng-l LFAKLVGNIAEDGGKLTDYLISHSALQPYQAGKSGYAAVQNGRYVLEIDSEGAFYFRRRHY 181 The complete length ORF108ng nucleotide sequence <SEQ ID 429> is: 1 ATGCTGAAAa tacctTTTGC CGTGTtgggc ggCtgcctGC TGCTTGCCGC 51 CTGCGGCAAA TCCGAAAATa cggcggaACA GCCGCAAAAT gcggCACAAA 101 GCGCGCCGAA ACCGGTTTTC AAAGTCAAAT ACATCGACAA TACGGCGATT 151 GCCGGTTTGG CTTTGGGACA AAGTAGCGAA GGCAAAACCA acgacgGCAA 201 AAAACAAATC AGTTATccgA TTAAAGGCTT GCCGGAACAA Aacgccgtcc 251 gGCTGACCGG AAAGCATCCC AACGACTTGG AagccgtcgT CGGCAAATGT 301 ATGGAAACCG ACGGAAAGGA CGCGCCTTCG GGCTGGGCGG AAAACGGCGT 351 GTGCCATACC TTGTTTGCCA AACTGGTGGG CAATATCGCC GAAGACGGCG 401 GCAAACTGAC TGATTACCTG ATTTCGCATT CCGCCCTGCA ACCCTATCAG 451 GCAGGCAAAA GCGGCTATGC CGCCGTGCAG AACGGACGCT ATGTGCTGGA 501 AATCGACAGC GagggGGCGT TTTATttccg ccgccgccat tattgA This encodes a protein having amino acid sequence <SEQ ID 430>: 1 MLKIPFAVLG GCLLLAACGK SENTAEQPQN AAQSAPKPVF KVKYIDNTAI 51 AGLALGQSSE GKTNDGKKQI SYPIKGLPEQ NAVRLTGKHP NDLEAWGKC 101 METDGKDAPS GWAENGVCHT LFAKLVGNIA EDGGKLTDYL ISHSALQPYQ 151 AGKSGYAAVQ NGRYVLEIDS EGAFYFRRRH Y* Based on this analysis, including the presence of a predicted prokaryotic membrane lipoprotein lipid attachment site (underlined) and a putative ATP/GTP-binding site motif A (P-loop, double- underlined) in the gonococcal protein, it is predicted that the proteins from N. meningitidis and N.gonorrhoeae, and their epitopes, could be useful antigens for vaccines or diagnostics, or for raising antibodies. Example 51 The following DNA sequence was identified in N. meningitidis <SEQ LD 431>: 1 ATGGAAGATT TATATATAAT ACTCGCTTTG GGTTTGGTTG CGATGATTGC 51 CGgATTTATC GATgcgatTg cGggCGGGGG TGGTTTGATT ACGCTGCCCG 101 CACTCTTGTT GGCAGGTATT CCTCCCGTGT CGGCAATTGC CACCAACAAG 151 CTGCAAgCAG CCGCTGCTAC GTTTTCAGCT ACGGTTTCTT TTGCACGCAA 201 AGGTTTGATT GATTGGAAGA AAGGTCTCCC GATTGCCGCA GCATCGTTTG 251 TAGGCGGCGT GGcCGGTGCA TTATCGGTCA GCTTGGTTTC CAAAGATATT 301 CTgCTgGCGG TCGTGCCGGT TTTGTTGATA TTTGTCGCAC TGTATTTTGT 351 GTTTTCGCCC AAGCTCGACG GCAGTAAGGA AGGCAAAGCC AGAATGTCTT 401 TTTTTCTGTT cGGGCTGACG GTCGC.ACCG CTTTTGGGTT TTTACGACGG 451 TGTGTTCGGA CCGGGTGTCG GCTCGTTTTT TCTGATTGCC TTTATTGTTT 501 TGCTCGGCTG CAAgCTGTTG AACGCGATGT CTTACACCAA ATTGGCGAAC 551 GTTGCCTGCA ATCTTGGTTC GCTATCGGTA TTCCTGCTGC ACGGTTCGAT 601 TATTTTCCCG ATTGCGGCAA CGaTGGCGGT CGGTGCGTTT GTCGGtGCGA 651 ATTTAgGTGC GAGATTTGCC GTaCgctTCG GTTCGAAGCT GATTAA This coπesponds to the amino acid sequence <SEQ ID 432; ORF109>: 1 MEDLYIILAL GLVAMIAGFI DAIAGGGGLI TLPALLLAGI PPVSAIATNK 51 LQAAAATFSA TVSFARKGLI DWKKGLPIAA ASFVGGVAGA LSVSLVSKDI 101 LLAWPVLLI FVALYFVFSP KLDGSKEGKA RMSFFLFGLT VXTAFGFLRR 151 CVRTGCRLVF SDCLYCFARL QAVERDVLHQ IGERCLQSWF AIGIPAARFD 201 YFPDCGNDGG RCVCRCEFRC EICRTLRFEA D* Further work revealed the following DNA sequence <SEQ ID 433>: 1 ATGGAAGATT TATATATAAT ACTCGCTTTG GGTTTGGTTG CGATGATTGC 51 CGGATTTATC GATGCGATTG CGGGCGGGGG TGGTTTGATT ACGCTGCCCG 101 CACTCTTGTT GGCAGGTATT CCTCCCGTGT CGGCAATTGC CACCAACAAG 151 CTGCAAGCAG CCGCTGCTAC GTTTTCAGCT ACGGTTTCTT TTGCACGCAA 201 AGGTTTGATT GATTGGAAGA AAGGTCTCCC GATTGCCGCA GCATCGTTTG 251 TAGGCGGCGT GGCCGGTGCA TTATCGGTCA GCTTGGTTTC CAAAGATATT 301 CTGCTGGCGG TCGTGCCGGT TTTGTTGATA TTTGTCGCAC TGTATTTTGT 351 GTTTTCGCCC AAGCTCGACG GCAGTAAGGA AGGCAAAGCC AGAATGTCTT 401 TTTTTCTGTT CGGGCTGACG GTCGCACCGC TTTTGGGTTT TTACGACGGT 451 GTGTTCGGAC CGGGTGTCGG CTCGTTTTTT CTGATTGCCT TTATTGTTTT 501 GCTCGGCTGC AAGCTGTTGA ACGCGATGTC TTACACCAAA TTGGCGAACG 551 TTGCCTGCAA TCTTGGTTCG CTATCGGTAT TCCTGCTGCA CGGTTCGATT 601 ATTTTCCCGA TTGCGGCAAC GATGGCGGTC GGTGCGTTTG TCGGTGCGAA 651 TTTAGGTGCG AGATTTGCCG TCCGCTTCGG TTCGAAGCTG ATTAAGCCGC 701 TGCTGATTGT CATCAGCATT TCGATGGCTG TGAAATTGTT GATAGACGAG 751 AGAAATCCGC TGTATCAGAT GATTGTTTCG ATGTTTTAA This coπesponds to the amino acid sequence <SEQ ID 434; ORF109-1>: 1 MEDLYIILAL GLVAMIAGFI DAIAGGGGLI TLPALLLAGI PPVSAIATNK 51 LQAAAATFSA TVSFARKGLI DWKKGLPIAA ASFVGGVAGA LSVSLVSKDI 101 LLAWPVLLI FVALYFVFSP KLDGSKEGKA RMSFFLFGLT VAPLLGFYDG 151 VFGPGVGSFF LIAFIVLLGC KLLNAMSYTK LANVACNLGS LSVFLLHGSI_ 201 IFPIAATMAV GAFVGANLGA RFAVRFGSKL IKPLLIVISI SMAVKLLIDE 251 RNPLYQMIVS MF* Computer analysis of this amino acid sequence gave the following results: Homology with a predicted ORF from N. meningitidis (strain A) ORF 109 shows 95.9% identity over a 147aa overlap with an ORF (ORF 109a) from sfrain A of N. meningitidis: 10 20 30 40 50 60 orf109. pep MEDLYIILALGLVAMIAGFIDAIAGGGGLITLPALLLAGIPPVSAIATNKLQAAAATFSA I II II II I II I I I I I I I I I II I I I II II II II I I I II I I I I I I II II I II I I I I II I I II orf109a MEDLYIILALGLVAMIAGFIDAIAGGGGLITLPALLLAGIPPVSAIATNKLQAAAATFSA 10 20 30 40 50 60 70 80 90 100 110 120 orf 109 . pep TVSFARKGLIDWKKGLPIAAASFVGGVAGALSVSLVSKDILLAWPVLLIFVALYFVFSP I I I I I I I I I I I I I I I I I I I I I I I : I I M I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I orf 109a TVSFARKGLIDWKKGLPIAAASFAGGWGALSVSLVSKDILLAWPVLLIFVALYFVFSP 70 80 90 100 110 120 130 140 150 160 170 180 orf 109. pep KLDGSKEGKARMSFFLFGLTVXTAFGFLRRCVRTGCRLVFSDCLYCFARLQAVERDVLHQ I II I I I II I II II II II II II Ml orfl09a KLDGSKEGKARMSFFLFGLTVAPLLGFYDGVFGPGVGSFFLIAFIVLLGCKLLNAMSYTK 130 140 150 160 170 180 The complete length ORF 109a nucleotide sequence <SEQ ID 435> is: 1 ATGGAAGATT TATACATAAT ACTCGCTTTG GGTTTGGTTG CGATGATTGC 51 CGGATTTATC GATGCGATTG CGGGTGGGGG TGGTTTGATT ACGCTGCCTG 101 CACTCTTGTT GGCAGGTATT CCTCCCGTGT CGGCAATTGC CACCAACAAG 151 CTGCAAGCAG CCGCTGCTAC GTTTTCGGCT ACGGTTTCTT TTGCACGCAA 201 AGGTTTGATT GATTGGAAGA AAGGTCTCCC GATTGCGGCA GCATCGTTTG 251 CAGGCGGCGT GGTCGGTGCA TTATCGGTCA GCTTGGTTTC CAAAGATATT 301 CTGCTGGCGG TCGTGCCGGT TTTGTTGATA TTTGTCGCGC TGTATTTTGT 351 GTTTTCGCCC AAGCTCGACG GCAGTAAGGA AGGCAAAGCC AGAATGTCTT 401 TTTTTCTGTT CGGTCTGACG GTTGCACCAC TTTTGGGTTT TTACGACGGT 451 GTGTTCGGAC CGGGTGTCGG CTCGTTTTTT CTGATTGCCT TTATTGTTTT 501 GCTCGGCTGC AAGCTGTTGA ACGCGATGTC TTACACCAAA TTGGCGAACG 551 TTGCCTGCAA TCTTGGTTCG CTATCGGTAT TCCTGCTGCA CGGTTCGATT 601 ATTTTCCCGA TTGCGGCAAC GATGGCGGTC GGTGCGTTTG TCGGTGCGAA 651 TTTAGGTGCG AGATTTGCCG TCCGCTTCGG TTCGAAGCTG ATTAAGCCGC 701 TGCTGATTGT CATCAGCATT TCGATGGCTG TGAAATTGTT GATAGACGAG 751 AGAAATCCGC TGTATCAGAT GATTGTTTCG ATGTTTTAA This encodes a protein having amino acid sequence <SEQ ID 436>: 1 MEDLYIILAL GLVAMIAGFI DAIAGGGGLI TLPALLLAGI PPVSAIATNK 51 LQAAAATFSA TVSFARKGLI DWKKGLPIAA ASFAGGWGA LSVSLVSKDI 101 LLAWPVLLI FVALYFVFSP KLDGSKEGKA RMSFFLFGLT VAPLLGFYDG 151 VFGPGVGSFF LIAFIVLLGC KLLNAMSYTK LANVACNLGS LSVFLLHGSI_ 201 IFPIAATMAV GAFVGANLGA RFAVRFGSKL IKPLLIVISI SMAVKLLIDE 251 RNPLYQMIVS MF* ORF109a and ORF109-1 show 99.2% identity in 262 aa overlap: 10 20 30 40 50 60 orf109a. pep MEDLYIILALGLVAMIAGFIDAIAGGGGLITLPALLLAGIPPVSAIATNKLQAAAATFSA I II I II II I II I II II II I I I II I I II I II II I I I II II II II II I II I I I II II II I II orf109-1 MEDLYIILALGLVAMIAGFIDAIAGGGGLITLPALLLAGIPPVSAIATNKLQAAAATFSA 10 20 30 40 50 60 70 80 90 100 110 120 orf109a. pep TVSFARKGLIDWKKGLPIAAASFAGGWGALSVSLVSKDILLAWPVLLIFVALYFVFSP I II I II II I II I I I II II I I I MM IM I I I I I II II II II II II I II I I II II II I I II orf109-1 TVSFARKGLIDWKKGLPIAAASFVGGVAGALSVSLVSKDILLAWPVLLIFVALYFVFSP 70 80 90 100 110 120 130 140 150 160 170 180 orf109a. pep KLDGSKEGKARMSFFLFGLTVAPLLGFYDGVFGPGVGSFFLIAFIVLLGCKLLNAMSYTK I I I I I II I I I II I II I I I I I II I I I II II II I I II II II I I II II I I I I I I I I II II I I I orf109-1 KLDGSKEGKARMSFFLFGLTVAPLLGFYDGVFGPGVGSFFLIAFIVLLGCKLLNAMSYTK 130 140 150 160 170 180 190 200 210 220 230 240 orf109a. pep LANVACNLGSLSVFLLHGSIIFPIAATMAVGAFVGANLGARFAVRFGSKLIKPLLIVISI I II I II II I I I I I I I I I I II I I I I II II I II II I I I I I I I I II II I I I I I II II II I I II orf109-1 LANVACNLGSLSVFLLHGSIIFPIAATMAVGAFVGANLGARFAVRFGSKLIKPLLIVISI 190 200 210 220 230 240 250 260 orf109a. pep SMAVKLLIDERNPLYQMIVSMFX I I II II I II I II I II I I I I I II I orfl09-l SMAVKLLIDERNPLYQMIVSMFX 250 260 Homology with a predicted ORF from N.εonorrhoeae ORF109 shows 98.3% identity over a 231aa overlap with a predicted ORF (ORF109.ng) fromN gonorrhoeae: orf 109 . pep MEDLYI ILALGLVAMIAGFIDAIAGGGGLITLPALLLAGIPPVSAIATΝKLQAAAATFSA 60 I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I orf l 09ng MEDLYIILALGLVAMIAGFIDAIAGGGGLITLPALLLAGIPPVSAIATΝKLQAAAATFSA 60 orf 109 . pep TVSFARKGLIDWKKGLPIAAASFVGGVAGALSVSLVSKDILLAWPVLLIFVALYFVFSP 120 I I I I I I I I I I I I I I I I I I I I I I I : I I I : I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I orfl09ng TVSFARKGLIDWKKGLPIAAASFAGGWGALSVSLVSKDILLAWPVLLIFVALYFVFSP 120 orf109. pep KLDGSKEGKARMSFFLFGLTVXTAFGFLRRCVRTGCRLVFSDCLYCFARLQAVERDVLHQ 180 I II I II II II I I I I I I II I I I I II II II II I I II II I I II I I I I I I II I I II I I I II II orfl09ng KLDGSKEGKARMSFFLFGLTVATAFGFLRRCVRTGCRLVFSDCLYCFARLQAVERDVLHQ 180 orf109. pep IGERCLQSWFAIGIPAARFDYFPDCGΝDGGRCVCRCEFRCEICRTLRFEAD 231 II I I I I I I II I I I I I I I I I I II I II I I I I II I I II II I I II II I I II I I I orfl09ng IGERCLQSWFAIGIPAARFDYFPDCGΝDGGRCVCRCEFRCEICRPLRFEAD 231 An ORF109ng nucleotide sequence <SEQ ED 437> was predicted to encode a protein having amino acid sequence <SEQ LD 438>: 1 MEDLYIILAL GLVAMIAGFI DAIAGGGGLI TLPALLLAGI PPVSAIATNK 51 LQAAAATFSA TVSFARKGLI DWKKGLPIAA ASFAGGWGA LSVSLVSKDI 101 LLAWPVLLI FVALYFVFSP KLDGSKEGKA RMSFFLFGLT VATAFGFLRR 151 CVRTGCRLVF SDCLYCFARL QAVERDVLHQ IGERCLQSWF AIGIPAARFD 201 YFPDCGNDGG RCVCRCEFRC EICRPLRFEA D* Further work revealed the following gonococcal DNA sequence <SEQ ID 439>: 1 ATGGAAGATT TATACATAAT ACTCGCTTTG GGTTTGGTTG CGATGATCGC 51 CGGATTTATC GATGCGATTG CGGGCGGGGG TGGTTTGATT ACGCTGCCTG 101 CACTCTTGTT GGCAGGTATT CCTCCCGTGT CGGCAATTGC CACCAACAAG 151 CTGCAAGCAG CCGCTGCTAC GTTTTCGGCT ACGGTTTCTT TTGCACGCAA 201 AGGTTTGATT GATTGGAAGA AAGGTCTCCC GATTGCCGCA GCATCGTTTG 251 CAGGCGGCGT GGTCGGTGCA TTATCGGTCA GCTTGGTTTC CAAAGATATT 301 TTGCTGGCGG TCGTGCCGGT TTTGTTGATA TTTGTCGCGC TGTATTTTGT 351 GTTTTCGCCC AAGCTCGACG GCAGTAAGGA AGGCAAAGCC AGAATGTCTT 401 TTTTTCTATT CGGGCTGACG GTTGCACCGC TTTTGGGTTT TTACGACGGT 451 GTGTTCGGAC CGGGTGTCGG CTCGTTTTTT CTGATTGCCT TTATTGTTTT 501 GCTCGGCTGC AAGCTGTTGA ACGCGATGTC TTACACCAAA TTGGCGAACG 551 TTGCTTGCAA TCTTGGTTCG CTATCGGTAT TCCTGCTGCA CGGTTCGATT 601 ATTTTCCCGA TTGTGGCAAC GATGGCGGTC GGTGCGTTTG TCGGTGCGAA 651 TTTAGGTGCG AGATTTGCCG TCCGCTTCGG TTCGAAGCTG ATTAAGCCGC 701 TGCTGATTGT CATCAGCATT TCGATGGCTG TGAAATTGTT GATAGACGAG 751 AGAAATCCGC TGTATCAGAT GATTGTTTCG ATGTTTTAA This coπesponds to the amino acid sequence <SEQ ID 440; ORF109ng-l>: 1 MEDLYIILAL GLVAMIAGFI DAIAGGGGLI TLPALLLAGI PPVSAIATNK 51 LQAAAATFSA TVSFARKGLI DWKKGLPIAA ASFAGGWGA LSVSLVSKDI 101 LLAWPVLLI FVALYFVFSP KLDGSKEGKA RMSFFLFGLT VAPLLGFYDG 151 VFGPGVGSFF LIAFIVLLGC KLLNAMSYTK LANVACNLGS LSVFLLHGSI ^ 201 IFPIVATMAV GAFVGANLGA RFAVRFGSKL IKPLLIVISI SMAVKLLIDE 251 RNPLYQMIVS MF* ORF109ng-l and ORF109-1 show 98.9% identity in 262 aa overlap: 10 20 30 40 50 60 orfl09ng-l.pep MEDLYIILALGLVAMIAGFIDAIAGGGGLITLPALLLAGIPPVSAIATNKLQAAAATFSA I I I I II I II I II I I II I II I I I II I II I I II I II I I I I II I I I I I I I I I II I I I I I II I I orf109-1 MEDLYIILALGLVAMIAGFIDAIAGGGGLITLPALLLAGIPPVSAIATNKLQAAAATFSA 10 20 30 40 50 60 70 80 90 100 110 120 orfl09ng-l.pep TVSFARKGLIDWKKGLPIAAASFAGGVVGALSVSLVSKDILLAWPVLLIFVALYFVFSP I II I I I II I I II II I I I I I I I I I: I I I : I I I I II I I II I I II II II I I I I I I II I I I II I orfl09-l TVSFARKGLIDWKKGLPIAAASFVGGVAGALSVSLVSKDILLAWPVLLIFVALYFVFSP 70 80 90 100 110 120 130 140 150 160 170 180 orfl09ng-l.pep KLDGSKEGKARMSFFLFGLTVAPLLGFYDGVFGPGVGSFFLIAFIVLLGCKLLNAMSYTK II I I I II I II II I I I I I I I I I I I II I I I I I I I II I I II I I II I I II I I I I II I II I I I II orf109-1 KLDGSKEGKARMSFFLFGLTVAPLLGFYDGVFGPGVGSFFLIAFIVLLGCKLLNAMSYTK 130 140 150 160 170 180 190 200 210 220 230 240 orfl09ng-l.pep LANVACNLGSLSVFLLHGSIIFPIVATMAVGAFVGANLGARFAVRFGSKLIKPLLIVISI I I II II I I I I I II II II I I I I II I : I I I I I II I II I I I I II I II I I I I I II I I I II I I I I orf109-1 LANVACNLGSLSVFLLHGSIIFPIAATMAVGAFVGANLGARFAVRFGSKLIKPLLIVISI 190 200 210 220 230 240 250 260 orf109ng-l .pep SMAVKLLIDERNPLYQMIVSMFX I II II I II I II I I I I I I II I II I orf109-1 SMAVKLLIDERNPLYQMIVSMFX 250 260 In addition, ORF109ng-l shows homology to a hypothetical Pseudomonas protein: sp|P29942|YCB9_PSEDE HYPOTHETICAL 27.4 KD PROTEIN IN COBO 3'REGION (ORF9) >gi I 94984 | pir | 1138164 hypothetical protein 9 - Pseudomonas sp >gi|551929 (M62866) ORF9 [Pseudomonas denitrificans] Length = 261 Score = 175 bits (439), Expect = 3e-43 Identities = 83/214 (38%), Positives = 131/214 (60%), Gaps = 1/214 (0%) Query: 41 PPVSAIATNKLQXXXXXXXXXXXXXRKGLIDWKKGLPIXXXXXXXXXXXXXXXXXXXKDI 100 PP+ + TNKLQ R+G ++ K+ LP+ D+ Sbjct: 43 PPLQTLGTNKLQGLFGSGSATLSYARRGHVNLKEQLPMALMSAAGAVLGALLATIVPGDV 102 Query: 101 LLA PVLLIFVALYFVFSPKLDGSKEGKARMSFFLFGLTVAPLLGFYDGVFGPGVGSFF 160 L A++P LLI +ALYF P + G + +R++ F+F LT+ PL+GFYDGVFGPG GSFF Sbjct: 103 LKAILPFLLIAIALYFGLKPNM-GDVDQHSRVTPFVFTLTLVPLIGFYDGVFGPGTGSFF 161 Query: 161 LIAFIVLLGCKLLNAMSYTKLANVACNLGSLSVFLLHGSIIFPIVATMAVGAFVGANLGA 220 ++ F+ L G +L A ++TK N N+G+ VFL G++++ + M +G F+GA +G+ Sbjct: 162 MLGFVTLAGFGVLKATAHTKFLNFGSNVGAFGVFLFFGAVLWKVGLLMGLGQFLGAQVGS 221 Query: 221 RFAVRFGSKLIKPLLIVISISMAVKLLIDERNPL 254 R+A+ G+K+IKPLL+++SI++A++LL D +PL Sbjct: 222 RYAMAKGAKIIKPLLVIVSIALAIRLLADPTHPL 255 Based on this analysis, including the presence of a putative leader sequence (double-underlined) and several putative transmembrane domains (single-underlined) in the gonococcal protein, it is predicted that the proteins from N. meningitidis .and N.gonorrhoeae, and their epitopes, could be useful antigens for vaccines or diagnostics, or for raising antibodies. Example 52 The following partial DNA sequence was identified in N. meningitidis <SEQ ED 441>: 1 .. CTGCTAGGGT ATTGCATCGG TTATCGGTAC GGCTGTTGCA GCAAAACCAG 51 CCGCAGACGG ATTATTTGGT CAAATTCGGA TCGTTTTGGG CGAG.ATTTT 101 TGGTTTTCTG GGACTGTATG ACGTCTATGC TTCGGCATGG TTTGTCGTTA 151 TCATGATGTT TTTGGTGGTT TCTACCAGTT TGTGCCTGAT TCGCAATGTG 201 CCGCCGTTCT GGCGCGAAAT GAAGTCTTTT CGGGAAAAGG TTAAAGAAAA 251 ATCTCTGGCG GCGATGCGCC ATTCTTCGCT GTTGGATGTA AAAATTGCGC 301 CCGAGGTTGC CAAACGTTAT CTGGAAGTAC AAGGTTTTCA GGGGAAAACC 351 ATTAACCGTG AAGACGGGTC GGTTCTGATT GCCGCCAAAA AAGGCACAAT 401 GAACAAATGG GGCTATATCT TTGCCCATGT TGCTTTGATT GTCATTTGCC 451 TGGGCGGGTT GATAGACAGT AACCTGCTGT TGAAACTGGG TATGCTGACC 501 GGTCGGATTG TTCCGGACAA TCAGGCGGTT TATGCCAAGG ATTTC . AAGC 551 CCGAAAGTAT . TTTGGGTGC gTCCAATCTC TCATTTAGGG GCAACGTCAA 601 TATTTCCG.A GGGGCAGAgT GCGGATGTGG TTTTCCTGA This coπesponds to the amino acid sequence <SEQ ID 442; ORFl 10>: 1 ..LLGIASVIGT LLQQNQPQTD YLVKFGSFWA XIFGFLGLYD VYASAWFWI 51 MMFLWSTSL CLIRNVPPFW REMKSFREKV KEKSLAAMRH SSLLDVKIAP 101 EVAKRYLEVQ GFQGKTINRE DGSVLIAAKK GTMNKWGYIF AHVALIVICL 151 GGLIDSNLLL KLGMLTGRIF RTIRRFMPRI XKPESXFGCV QSLI*GQRQY 201 FXRGRVRMWF S* Computer analysis of this amino acid sequence gave the following results: Homology with ORF88a from N.meninsitidis (sfrain Al ORFl 10 shows 91.5% identity over a 188aa overlap with ORF88a from strain A of N meningitidis: 10 20 30 40 50 60 orf88a . pep MSKSRRSPPLLSRPWFAFFSSMRFAVALLSLLGIASVIGTVLQQΝQPQTDYLVKFGSFWA II II I II II I : I I I II I I I I I II II I II I I orfllO LLGIASVIGTLLQQΝQPQTDYLVKFGSFWA 10 20 30 70 80 90 100 110 120 orf88a.pep QIFGFLGLYDVYASAWFWIMMFLWSTSLCLIRΝVPPFWREMKSFREKVKEKSLAAMRH I I II I II II I I II I I II II II I II II I II II I II I I I II II I II I I I I I I I II II I II I orfll0 XIFGFLGLYDVYASAWFWIMMFLWSTSLCLIRΝVPPFWREMKSFREKVKEKSLAAMRH 40 50 60 70 80 90 130 140 150 160 170 180 orf88a . pep SSLLDVKIAPEVAKRYLEVQGFQGKTIΝREDGSVLIAAKKGTMΝKWGYIFAHVALIVICL I II I II I II I I I I I I I I I I I I I I I I I I I I I I I II I I I I I I II I II I II I I II I I II II I I orf110 SSLLDVKIAPEVAKRYLEVQGFQGKTIΝREDGSVLIAAKKGTMΝKWGYIFAHVALIVICL 100 110 120 130 140 150 190 200 210 220 230 240 orf88a. pep GGLIDSΝLLLKLGMLTGRIVPDΝQAVYAKDFKPESILGASΝLSFRGΝVΝISEGQSADWF II I II I II II I II II II I I : : : MM M orf110 GGLIDSΝLLLKLGMLTGRIFRTIRRFMPRIXKPESXFGCVQSLIXGQRQYFXRGRVRMWF 160 170 180 190 200 210 250 260 270 280 290 300 orf88a . pep LΝADΝGILVQDLPFEVKLKKFHIDFYΝTGMPRDFASDIEVTDKATGEKLERTIRVΝHPLT orfllO SX However, ORF88 and ORFl 10 do not align, because they represent two different fragments of the same protein. Homology with a predicted ORF from N.εonorrhoeae ORFl 10 shows 88.6% identity over a 21 laa overlap with a predicted ORF (ORFl lO.ng) from N gonorrhoeae: orf110. pep LLGIASVIGTLLQQΝQPQTDYLVKFGSFWA 30 II I I II I I I I : I I I I I I I I I II I I I I I I : orfllOng MSKSRISPTLLSRPWFAFFSSMRFAVALLSLLGIASVIGTVLQQΝQPQTDYLVKFGPFWT 60 orfllO . pep XIFGFLGLYDVYASAWFWIMMFLWSTSLCLIRΝVPPFWREMKSFREKVKEKSLAAMRH 90 M II I I I II I I I II II II I II I I I II II I II I I II II I II I I I I II II I I I II I I I I I orfllOng RIFDFLGLYDVYASAWFWIMMFLWSTSLCLIRΝVPPFWREMKSFREKVKEKSLAAMRH 120 orfllO . pep SSLLDVKIAPEVAKRYLEVQGFQGKTIΝREDGSVLIAAKKGTMΝKWGYIFAHVALIVICL 150 II I II II II II I II I I I I I: I II I I M: I I I I II I II I II I II II I I II I II II I I II I orfllOng SSLLDVKIAPEVAKRYLEVRGFQGKTVSREDGSVLIAAKKGTMΝKWGYIXAHVALIVICL 180 orfllO.pep GGLIDSNLLLKLGMLTGRIFRTIRRFMPRIXKPESXFGCVQSLIXGQRQYFXRGRVRMWF 210 I II: I I I I II II I : I MM II II II I II I M II I II I I I I II II : I I I I I orfllOng GRLINXNLLLKLGMLAGSIFRNNRRVMPRISKPESIWGGVQSLIKGQRQYFQRGKVRMWF 240 orfllO. pep S 211 I orfllOng S 241 The complete length ORF 11 Ong nucleotide sequence <SEQ LD 443> is predicted to encode a protein having amino acid sequence <SEQ ID 444>: 1 MSKSRISPTL LSRPWFAFFS SMRFAVALLS LLGIASVIGT VLQQNQPQTD 51 YLVKFGPFWT RIFDFLGLYD VYASAWFWI MMFLVVSTSL CLIRNVPPFW 101 REMKSFREKV KEKSLAAMRH SSLLDVKIAP EVAKRYLEVR GFQGKTVSRE 151 DGSVLIAAKK GTMNKWGYIX AHVALIVICL GRLINXNLLL KLGMLAGSIF 201 RNNRRVMPRI SKPESIWGGV QSLIKGQRQY FQRGKVRMWF S* Based on the putative transmembrane domains in the gonococcal protein, it is predicted that the proteins from N. meningitidis and N.gonorrhoeae, and their epitopes, could be useful antigens for vaccines or diagnostics, or for raising antibodies. Example 53 The following DNA sequence was identified in N. meningitidis <SEQ ID 445>: 1 ATGCCGTCTG AAACACGCCT GCCGAACTTT ATCCGCGTCT TGATATTTGC 51 CCTGGGTTTC ATCTTCCTGA ACGCCTGTTC GGAACAAACC GCGCAAACCG 101 TTACCCTGCA AGGCGAAACG ATGGGCACGA CCTATACCGT CAAATACCTT 151 TCAAATAATC GGGACAAACT CCCCTCACCT GCCGAAATAC AAAAACGCAT 201 CGATGACGCG CTTAAAGAAG TCAACCGGCA GATGTCCACC TATCAGCCCG 251 ACTCCGAAAT CAGCCGGTTC AACCAACACA CAGCCGGCAA GCCCCTCCGC 301 ATTTCAAGCG ACTTCGCACA CGTTACTGCC GAAGCCGTCC GCCTGAACCG 351 CCTGACACAC GGCGCGCTGG ACGTAACCGT CGGCCCCTTG GTCAACCTTT 401 GGGGATTCGG CCCCGACAAA TCCGTTACCC GTGAACCGTC GCCGGAACAA 451 ATCAAACAGG CGGCATCTTA TACGGGCATA GACAAAATCA TTTTGAAACA 501 AGGCAAAGAT TACGCTTCCT TGAGCAAAAC CCACCCCAAG GCCTATTTGG 551 ATTTATCTTC GATTGCCAAA GGCTTCGGCG TTGATAAAGT TGCGGGCGAA 601 CTGGAAAAAT ACGGCATTCA AAATTATCTG GTCGAAATCG GCGGCGAGTT 651 GCACGGCAAA GGCAAAAACG CGCGCGGCGA ACCGTGGCGC ATCGGTATCG 701 AGCAGCCCAA TATCGTCCAA GGCGGCAATA CGCAGATTAT CGTCCCGCTG 751 AACAACCGTT CGCTTGCCAC TTCCGGCGAT TACCGTATTT TCCACGTCGA 801 TAAAAACGGC AAACGCCTCT CCCATATCAT CAACCCGAAC AACAAACGAC 851 CCATCAGCCA CAACCTCGCC TCCATCAGCG TGGTCGCAGA CAGTGCGATG 901 ACGGCGGACG GCTTGTCCAC AGGATTATTC GTATTGGGCG AAACCGAAGC 951 CTTAAAGCTG GCAGAGCGCG AAAAACTCGC TGTTTTCCTG ATTGTCAGGG 1001 ATAAAGGCGG CTACCGCACC GCCATGTCTT CCGAATTTGA AAAACTGCTC 1051 CGCTAA This coπesponds to the amino acid sequence <SEQ ID 446; ORFl 11>: 1 MPSETRLPNF IRVLIFALGF IFLNACSEQT AQTVTLQGET MGTTYTVKYL 51 SNNRDKLPSP AEIQKRIDDA LKEVNRQMST YQPDSEISRF NQHTAGKPLR 101 ISSDFAHVTA EAVRLNRLTH GALDVTVGPL VNLWGFGPDK SVTREPSPEQ 151 IKQAASYTGI DKIILKQGKD YASLSKTHPK AYLDLSSIAK GFGVDKVAGE 201 LEKYGIQNYL VEIGGELHGK GKNARGEPWR IGIEQPNIVQ GGNTQIIVPL 251 NNRSLATSGD YRIFHVDKNG KRLSHIINPN NKRPISHNLA SISWADSAM 301 TADGLSTGLF VLGETEALKL AEREKLAVFL IVRDKGGYRT AMSSEFEKLL 351 R* Computer analysis of this amino acid sequence gave the following results: Homology with a predicted ORF from N.meninsitidis (sfrain Al ORFl 11 shows 96.9% identity over a 351aa overlap with an ORF (ORFl 1 la) from strain A of N meningitidis: 10 20 30 40 50 60 orfIlia . pep MPSETRLPΝFIRTLIFALSFIFLΝACSEQTAQTVTLQGETMGTTYTVKYLSΝΝRDXLPSP I II II I I I II I I : I II II : I I I I II I I I I I II I I I II I I I II I I I I I II I I II I I I I II orfill MPSETRLPΝFIRVLIFALGFIFLΝACSEQTAQTVTLQGETMGTTYTVKYLSΝΝRDKLPSP 10 20 30 40 50 60 70 80 90 100 110 120 orfIlia. pep AEIQXRIDDALKEVΝRQMSTYQPDSEISRFΝQHTAGKPLRISSDFAHVTAEAVHLΝRLTH I II I I I I I II II II II I II I I II I I I II I I I II II I I I I I I I I I I II II I I I : I I I I I I orf111 AEIQKRIDDALKEVΝRQMSTYQPDSEISRFΝQHTAGKPLRISSDFAHVTAEAVRLΝRLTH 70 80 90 100 110 120 130 140 150 160 170 180 orfIlia . pep GALDVTVGPLVΝLWGFGPDKSVTREPSPEQIKQAASYTGIDKIILKQGKDYASLSKTHPK II II I II I II I I I I I I II I I I II I I I I I I II I II I I I I I I I I I I I I II I II I II II I I I I orf111 GALDVTVGPLVΝLWGFGPDKSVTREPSPEQIKQAASYTGIDKIILKQGKDYASLSKTHPK 130 140 150 160 170 180 190 200 210 220 230 240 orfIlia. pep AYLDLSSIAKGFGVDXVAGELEKYGIQΝYLVEIGGELHGKXKΝARGEPWRIGIEQPΝIVQ I II I I I II I II II II I I I I I I I II I II II I II II I II II II I I I I I II I II I I I I I II orf111 AYLDLSSIAKGFGVDKVAGELEKYGIQΝYLVEIGGELHGKGKΝARGEPWRIGIEQPΝIVQ 190 200 210 220 230 240 250 260 270 280 290 300 orfIlia. pep GGΝTQIIVPLΝΝRSXATSGDYRIFHVDKSGKRLSHIIΝPΝΝKRPISHΝLASISVXADSAM II II I II I II I I I I II I II II I I I II IM I I I II I II II I II II I I I II I II I I I I I I orf111 GGΝTQIIVPLΝΝRSLATSGDYRIFHVDKΝGKRLSHIIΝPΝΝKRPISHΝLASISWADSAM 250 260 270 280 290 300 310 320 330 340 350 orfIlia . pep TADGXSTGLFVLGETEALKLAEREKLAVFLIVRDKGGYRTAMSSEFEKLLRX I II I I I II II II I II I II I I II I I I I II I I I I II II I I I II I I I I I I I II I orfill TADGLSTGLFVLGETEALKLAEREKLAVFLIVRDKGGYRTAMSSEFEKLLRX 310 320 330 340 350 The complete length ORFl 1 la nucleotide sequence <SEQ ID 447> is: 1 ATGCCGTCTG AAACACGCCT GCCGAACTTT ATCCGCACCT TGATATTTGC 51 CCTGAGTTTT ATCTTCCTGA ACGCCTGTTC GGAACAAACC GCGCAAACCG 101 TTACCCTGCA AGGTGAAACG ATGGGCACGA CCTATACCGT CAAATACCTT 151 TCAAATAATC GGGACΝAACT CCCΝTCACCT GCCGAAATAC AAAAΝCGCAT 201 CGATGACGCG CTTAAAGAAG TCAACCGGCA GATGTCCACC TATCAGCCCG 251 ACTCCGAAAT CAGCCGGTTC AACCAACACA CAGCCGGCAA GCCCCTCCGC 301 ATTTCAAGCG ACTTCGCACA CGTTACTGCC GAAGCCGTCC ACCTGAACCG 351 CCTGACACAC GGCGCGCTGG ACGTAACCGT CGGCCCCTTG GTCAACCTTT 401 GGGGATTCGG CCCCGACAAA TCCGTTACCC GTGAACCGTC GCCGGAACAA 451 ATCAAACAAG CAGCATCTTA TACGGGCATA GACAAAATCA TTTTGAAACA 501 AGGCAAAGAT TACGCTTCCT TGAGCAAAAC CCACCCCAAG GCCTATTTGG 551 ATTTATCTTC GATTGCCAAA GGCTTCGGCG TTGATΝAΝGT TGCGGGCGAA 601 CTGGAAAAAT ACGGCATTCA AAATTATCTG GTCGAAATCG GCGGΝGAGTT 651 GCACGGCAAA GΝCAAAAACG CGCGCGGCGA ACCTTGGCGC ATCGGCATCG 701 AACAGCCCAA CATCGTCCAA GGCGGCAATA CGCAGATTAT CGTCCCGCTG 751 AACAACCGTT CGΝTTGCCAC TTCCGGCGAT TACCGTATTT TCCACGTCGA 801 TAAAAGCGGC AAACGCCTCT CCCATATCAT TAATCCGAAC AACAAACGAC 851 CCATCAGCCA CAACCTCGCC TCCATCAGCG TGΝTCGCAGA CAGTGCGATG 901 ACGGCGGACG GCTTΝTCCAC AGGATTATTC GTATTGGGCG AAACCGAAGC 951 CTTAAAGCTG GCAGAGCGCG AAAAACTCGC TGTTTTCCTG ATTGTCAGGG 1001 ATAAAGGCGG CTACCGCACC GCCATGTCTT CCGAATTTGA AAAACTGCTC 1051 CGCTAA This encodes a protein having amino acid sequence <SEQ ID 448>: 1 MPSETRLPΝF IRTLIFALSF IFLΝACSEQT AQTVTLQGET MGTTYTVKYL 51 SΝΝRDXLPSP AEIQXRIDDA LKEVΝRQMST YQPDSEISRF ΝQHTAGKPLR 101 ISSDFAHVTA EAVHLNRLTH GALDVTVGPL VNLWGFGPDK SVTREPSPEQ 151 IKQAASYTGI DKIILKQGKD YASLSKTHPK AYLDLSSIAK GFGVDXVAGE 201 LEKYGIQNYL VEIGGELHGK XKNARGEPWR IGIEQPNIVQ GGNTQIIVPL 251 NNRSXATSGD YRIFHVDKSG KRLSHIINPN NKRPISHNLA SISVXADSAM 301 TADGXSTGLF VLGETEALKL AEREKLAVFL IVRDKGGYRT AMSSEFEKLL 351 R* Homology with a predicted ORF from Nsonorrhoeae ORFl 11 shows 96.6% identity over a 35 laa overlap with a predicted ORF (ORFl 1 l.ng) from N gonorrhoeae: 10 20 30 40 50 60 orflllng MPSETRLPΝLIRALIFALGFIFLΝACSEQTAQTVTLQGETMGTTYTVKYLSΝΝRDKLPSP I II II II IM I I : II II II II I II I I I II II II I II I I I I I II I II I I I II I I II II I I I orf111 MPSETRLPΝFIRVLIFALGFIFLΝACSEQTAQTVTLQGETMGTTYTVKYLSΝΝRDKLPSP 10 20 30 40 50 60 70 80 90 100 110 120 orfll1 AKIQKRIDDALKEVΝRQMSTYQTDSEISRFΝQHTAGKPLRISSDFAHVTAEAVRLΝRLTH I: I I I II II I I II I I I I I I I II II I I I I I II I II I II I II II I I II II I I II I I I I I II orf111 AEIQKRIDDALKEVΝRQMSTYQPDSEISRFΝQHTAGKPLRISSDFAHVTAEAVRLΝRLTH 70 80 90 100 110 120 130 140 150 160 170 180 orflllng GALDVTVGPLVΝLWGFGPDKSVTREPSPEQIKQAASYTGIDKIILQQGKDYASLSKTHPK II II I I I II I I I I I II II II I I I I I I I II II II I I II II I I I II I : I I II II I I I I I II I orf111 GALDVTVGPLVΝLWGFGPDKSVTREPSPEQIKQAASYTGIDKIILKQGKDYASLSKTHPK 130 140 150 160 170 180 190 200 210 220 230 240 orflllng AYLDLSSIAKGFGVDKVAGELEKYGIQΝYLVEIGGELHGKGKΝAHGEPWRIGIEQPΝIIQ I I II I I II I II I I II II II II I I II I I I I I II I II I II II I II I : II I I II I I II II I : I orfll1 AYLDLSSIAKGFGVDKVAGELEKYGIQΝYLVEIGGELHGKGKΝARGEPWRIGIEQPΝIVQ 190 200 210 220 230 240 250 260 270 280 290 300 orflllng GGΝTQIIVPLΝΝRSLATSGDYRIFHVDKΝGKRLSHIIΝPΝΝKRPISHΝLASISWSDSAM II II I I II II I I I II I I I I I II I II I II II II I II I I I I II I I I I I I I I II I I II M I I I orf111 GGΝTQIIVPLΝΝRSLATSGDYRIFHVDKΝGKRLSHIIΝPΝΝKRPISHΝLASISWADSAM 250 260 270 280 290 300 310 320 330 340 350 orflllng TADGLSTGLFVLGETEALRLAEQEKLAVFLIVRDKDGYRTAMSSEFAKLLRX I II I I I I I II I I I I I I I I : I II : II I II II II I II I I I I I I I I I I I II II orf111 TADGLSTGLFVLGETEALKLAEREKLAVFLIVRDKGGYRTAMSSEFEKLLRX 310 320 330 340 350 The complete length ORFl 1 lng nucleotide sequence <SEQ ID 449> is: 1 ATGCCGTCTG AAACACGCCT GCCGAACCTT ATCCGCGCCT TGATATTTGC 51 CCTGGGTTTC ATCTTCCTGA ACGCCTGTTC GGaacaaacC GCGCAaaccg 101 TTACCCTGCA AGGCGAAAcg aTGGGTACGA CCTATACCGT CAAATACCTT 151 TCAAATAATC GGGACAAACT CCCCTCCCCT GCCAAAATAC AAAAGCGCAT 201 TGATGATGCG CTTAAAGAAG TCAACCGGCA GATGTCCACC TACCAGACCG 251 ATTCCGAAAT CAGCCGGTTC AACCAACACA CAGCCGGCAA GCCCCTCCGC 301 ATTTCAAGCG ATTTCGCACA CGTTACCGCC GAAGCCGTCC GCCTGAACCG 351 CCTGACTCAC GGCGCACTGG ACGTAACCGT CGGCCCTTTG GTCAACCTTT 401 GGGGGTTCGG CCCCGACAAA TCCGTTACCC GTGAACCGTC GCCGGAACAA 451 ATCAAACAGG CGGCATCTTA TACGGGCATA GACAAAATCA TTTTGCAACA 501 AGGCAAAGAT TACGCTTCCT TGAGCAAAAC CCACCCCAAA GCCTATTTGG 551 ATTTATCTTC GATTGCCAAA GGCTTCGGCG TTGATAAAGT TGCGGGCGAA 601 CTGGAAAAAT ACGGCATTCA AAATTATCTG GTCGAAAtcg gcggcGAGTT 651 GCACGGCAAA GGCAAAAATG CGCACGGCGA ACCGTGGCGC ATCGGTATAG 701 AGCAACCCAA TATCATCCAA GgcgGCAata CGCAGATTAt cgtcccgctg 751 aaCaaccgtt cgctTGCCAC TTCCGGCGAT TAccgtaTTT tccacgtcgA 801 TAAAAAcggc aaacgccttt cccacaTCAT CAATCCCaAC aacAAACgac 851 ccATCAGcca caacctcgcc tccatcagcg tggtctcAGA CAGTGCAATG 901 ACGGCGGACG GTTtatCCAC AGGATTATTT GTTTTAGGCG AAACCGAAGC 951 CTTAAGGCTG GCAGAACAAG AAAAACTCGC TGTTTTCCTA ATTGTCCGGG 1001 ATAAGGACGG CTACCGCACC GCCATGTCTT CCGAATTTGC CAAGCTGCTC 1051 CGCTAA This encodes a protein having amino acid sequence <SEQ ID 450>: 1 MPSETRLPNL IRALIFALGF IFLNACSEQT AQTVTLQGET MGTTYTVKYL 51 SNNRDKLPSP AKIQKRIDDA LKEVNRQMST YQTDSEISRF NQHTAGKPLR 101 ISSDFAHVTA EAVRLNRLTH GALDVTVGPL VNLWGFGPDK SVTREPSPEQ 151 IKQAASYTGI DKIILQQGKD YASLSKTHPK AYLDLSSIAK GFGVDKVAGE 201 LEKYGIQNYL VEIGGELHGK GKNAHGEPWR IGIEQPNIIQ GGNTQIIVPL 251 NNRSLATSGD YRIFHVDKNG KRLSHIINPN NKRPISHNLA SISWSDSAM 301 TADGLSTGLF VLGETEALRL AEQEKLAVFL IVRDKDGYRT AMSSEFAKLL 351 R* This protein shosw homology with a hypothetical lipoprotein precursor from H.influenzae: sp|P44550|YOJL_HAEIN HYPOTHETICAL LIPOPROTEIN HI0172 PRECURSOR >gι | 1074292 | pir | 4 hypothetical protein HI0172 - Haemophilus influenzae (strain Rd KW20) >gι 11573128 (U32702) hypothetical [Haemophilus influenzae] Length = 346 Score = 353 bits (896), Expect = 9e-97 Identities = 181/344 (52%), Positives = 247/344 (71%), Gaps = 4/344 (1%) Query: 7 LPNLIRALIFALGFIFLNACSEQTAQTVTLQGETMGTTYTVKYLSNNRDKLPSPAKIQKR 66 + LI +1 + L AC ++T + ++L G+TMGTTY VKYL + S K + Sbjct: 1 MKKLISGIIAVAMALSLAACQKET-KVISLSGKTMGTTYHVKYLDDGSITATSE-KTHEE 58 Query: 67 IDDALKEVNRQMSTYQTDSEISRFNQHT-AGKPLRISSDFAHVTAEAVRLNRLTHGALDV 125 1+ LK+VN +MSTY+ DSE+SRFNQ+T P+ IS+DFA V AEA+RLN++T GALDV Sbjct: 59 IEAILKDVNAKMSTYKKDSELSRFNQNTQVNTPIEISADFAKVLAEAIRLNKVTEGALDV 118 Query: 126 TVGPLVNLWGFGPDKSVTREPSPEQIKQAASYTGIDKIILQQGKDYASLSKTHPKAYLDL 185 TVGP+VNLWGFGP+K ++P+PEQ+ + ++ GIDKI L K+ A+LSK P+ Y+DL Sbjct: 119 TVGPWNLWGFGPEKRPEKQPTPEQLAERQAWVGIDKITLDTNKEKATLSKALPQVYVDL 178 Query: 186 SSIAKGFGVDKVAGELEKYGIQNYLVEIGGELHGKGKNAHGEPWRIGIEQPNIIQGGNTQ 245 SSIAKGFGVD+VA +LE+ QNY+VEIGGE+ KGKN G+PW+I IE+P + Sbjct: 179 SSIAKGFGVDQVAEKLEQLNAQNYMVEIGGEIRAKGKNIEGKPWQIAIEKPTTTGERAVE 238 Query: 246 IIVPLNNRSLATSGDYRIFHVDKNGKRLSHIINPNNKRPISHNLASISVVSDSAMTADGL 305 ++ LNN +A+SGDYRI+ ++NGKR +H I+P PI H+LASI+V++ ++MTADGL Sbjct: 239 AVIGLNNMGMASSGDYRIY-FEENGKRFAHEIDPKTGYPIQHHLASITVLAPTSMTADGL 297 Query: 306 STGLFVLGETEALRLAEQEKLAVFLIVRDKDGYRTAMSSEFAKL 349 STGLFVLGE +AL +AE+ LAV+LI+R +G+ T SS F KL Sbjct: 298 STGLFVLGEDKALEVAEKNNLAVYLIIRTDNGFVTKSSSAFKKL 341 Based on this analysis, it is predicted that the proteins from Nmeningitidis .and N.gonorrhoeae, and their epitopes, could be useful antigens for vaccines or diagnostics, or for raising antibodies. Example 54 The following partial DNA sequence was identified in N. meningitidis <SEQ ID 451>: 1 .. CCGTGCCGCC GACAGGGCGA CGACGTGTAT GCGGCGCACG CGTCCCGTCA 51 AAAATTGTGG CTGCGCTTCA TCGGCGGCCG GTCGCATCAA AATATACGGG 101 GCGGCGCGGC TGCGGACGGG TGGCGCAAAG GCGTGCAAAT CGGCGGCGAG 151 GTGTTTGTAC GGCAAAATGA AGGCAGCCkA yTGGCAATCG GCGTGATGGG 201 CGGCAGGGCC GGCCAGCACG CwTCAGTCAA CGGCAAAGGC GGTGCGGCAG 251 gCAGTGATTT GTATGGTTAT GgCGGGGgTG TTTATGCTgC GTGGCATCAG 301 TTGCGCGATA AACAAACGGG TgCGTATTTG GACGGCTGGT TGCAATACCA 351 ACGTTTCAAA CACCGCATCA ATGATGAAAA CCGTGCGGAA CgCTACAAAA 401 CCAAAGGTTG GACGGCTTCT GTCGAAGGCG GCTACAACGC GCTTGTGGCG 451 GAAGGCATTG TCGGAAAAGG CAATAATGTG CGGTTTTACC TACAACCGCA 501 GgCGCAGTTT ACCTACTTGG GCGTAAACGG CGGCTTTACC GACAGCGAGG 551 GGACGGCGGT CGGACTGCTC GGCAGCGGTC AGTGGCAAAG CCGCGCCGGC 601 AtTCGGGCAA AAACCCGTTT TGCTTTGCGT AACGGTGTCA ATCTTCAGCC 651 TTTTGCCGCT TTTAATGTtt TGCACAGGTC AAAATCTTTC GGCGTGGAAA 701 TGGACGGCGA AAAACAGACG CTGGCAGGCA GGACGGCACT CGAAGGGCGG 751 TTCGGTATTG AAGCCGGTTG GAAAGGCCAT ATGTCCGCA.. This coπesponds to the amino acid sequence <SEQ ID 452; ORF35>: 1 .. PCRRQGDDVY AAHASRQKLW LRFIGGRSHQ NIRGGAAADG WRKGVQIGGE 51 VFVRQNEGSX LAIGVMGGRA GQHASVNGKG GAAGSDLYGY GGGVYAAWHQ 101 LRDKQTGAYL DGWLQYQRFK HRINDENRAE RYKTKGWTAS VEGGYNALVA 151 EGIVGKGNNV RFYLQPQAQF TYLGVNGGFT DSEGTAVGLL GSGQWQSRAG 201 IRAKTRFALR NGVNLQPFAA FNVLHRSKSF GVEMDGEKQT LAGRTALEGR 251 FGIEAGWKGH MSA.. Computer analysis of this amino acid sequence gave the following results: Homology with putative secreted VirG-homolgue of N meninsitidis (accession number A32247) ORF and virg-h protein show 51% aa identity in 26 laa overlap: Orf35 5 QGDDVYAAHASRQKLWLRFIGGRSHQΝIRGGAA-ADGWRKGVQIGGEVFVRQΝEGSXLAI 63 + D++ R+ LWLR I G S+Q ++G A +G+RKGVQ+GGEVF QΝE + L+I virg-h 396 KΝSDIFDRTLPRKGLWLRVIDGHSΝQWVQGKTAPVEGYRKGVQLGGEVFTWQΝESΝQLSI 455 0rf35 64 GVMGGRAGQHASVΝGKG—GAAGSDLYGYGGGVYAAWHQLRDKQTGAYLDGWLQYQRFKH 121 G+MGG+A Q ++ + ++ G+G GVYA WHQL+DKQTGAY D W+QYQRF+H virg-h 456 GLMGGQAEQRSTFHΝPDTDΝLTTGΝVKGFGAGVYATWHQLQDKQTGAYADSWMQYQRFRH 515 Orf35 122 RIΝDEΝRAERYKTKGWTASVEGGYΝALVAEGIVGKGΝΝVRFYLQPQAQFTYLGVΝGGFTD 181 RIΝ E+ ER+ +KG TAS+E GYΝAL+AE KGΝ++R YLQPQAQ TYLGVNG F+D virg-h 516 RINTEDGTERFTSKGITASIEAGYNALLAEHFTKKGNSLRVYLQPQAQLTYLGVNGKFSD 575 0rf35 182 SEGTAVGLLGSGQWQSRAGIRAKTRFALRNGVNLQPFAAFNVLHRSKSFGVEMDGEKQTL 241 SE V LLGS Q Q+R G++AK +F+L + ++PFAA N L+ +K FGVEMDGE++ + virg-h 576 SENAHVNLLGSRQLQTRVGVQAKAQFSLYKNIAIEPFAAVNALYHNKPFGVEMDGERRVI 635 Orf35 242 AGRTALEGRFGIEAGWKGHMS 262 +TA+E + G+ K H++ virg-h 636 NNKTAIESQLGVAVKIKSHLT 656 Homology with a predicted ORF from N.meninsitidis (sfrain A) ORF35 shows 96.9% identity over a 259aa overlap with an ORF (ORF35a) from strain A of N meningitidis: 10 20 30 orf35.pep PCRRQGDDVYAAHASRQKLWLRFIGGRSHQΝIRG : I I I II I I I I II I I I I I I I I II I I II I I orf35a QRLAIPEAEAVLYAQQAYAAΝTLFGLRAADRGDDVYAADPSRQKLWLRFIGGRSHQΝIRG 310 320 330 340 350 360 40 50 60 70 80 90 orf35. pep GAAADGWRKGVQIGGEVFVRQΝEGSXLAIGVMGGRAGQHASVΝGKGGAAGSDLYGYGGGV II I II I II I I I I I II I I I II I I I I I I II II I I II I II I I II I II II II I I : I I I I II orf35a GAAADGRRKGVQIGGEVFVRQΝEGSRLAIGVMGGRAGQHASVΝGKGGAAGSYLHGYGGGV 370 380 390 400 410 420 100 110 120 130 140 150 orf35. pep YAAWHQLRDKQTGAYLDGWLQYQRFKHRIΝDEΝRAERYKTKGWTASVEGGYΝALVAEGIV II I II II II I I I II I I I II I II I I I II I II I II I I I II II I I I I I I I I I I I II II II M I orf35a YAAWHQLRDKQTGAYLDGWLQYQRFKHRIΝDEΝRAERYKTKGWTASVEGGYΝALVAEGW 430 440 450 460 470 480 160 170 180 190 200 210 orf35. pep GKGΝΝVRFYLQPQAQFTYLGVΝGGFTDSEGTAVGLLGSGQWQSRAGIRAKTRFALRΝGVΝ I I I II I I II II I I I I I I II I I II I I I II I II I II I I II II I I I II II I I I II II II II I I orf35a GKGΝΝVRFYLQPQAQFTYLGVΝGGFTDSEGTAVGLLGSGQWQSRAGIRAKTRFALRΝGVΝ 490 500 510 520 530 540 220 230 240 250 260 orf35. pep LQPFAAFΝVLHRSKSFGVEMDGEKQTLAGRTALEGRFGIEAGWKGHMSA I I II I I I I II I I I I II I II I II I I I II I II I II II I II II I I I II II I I orf35a LQPFAAFNVLHRSKSFGVEMDGEKQTLAGRTALEGRFGIEAGWKGHMSARIGYGKRTDGD 550 560 570 580 590 600 orf35a KEAALSLKWLFX 610 620 The complete length ORF35a nucleotide sequence <SEQ ID 453> is: 1 ATGTTCAGAG CTCAGCTTGG TTCAAATACT CGTTCTACCA AAATCGGCGA 51 CGATGCCGAT TTTTCATTTT CAGACAAGCC GAAACCCGGC ACTTCCCATT 101 ATTTTTCCAG CGGTAAAACC GATCAAAATT CATCCGAATA TGGGTATGAC 151 GAAATCAATA TCCAAGGTAA AAACTACAAT AGCGGCATAC TCGCCGTCGA 201 TAATATGCCC GTTGTTAAGA AATATATTAC AGATACTTAC GGGGATAATT 251 TAAAGGATGC GGTTAAGAAG CAATTACAGG ATTTATACAA AACAAGACCC 301 GAAGCTTGGG AAGAAAATAA AAAACGGACT GAGGAGGCGT ATATAGAACA 351 GCTTGGACCA AAATTTAGTA TACTCAAACA GAAAAACCCC GATTTAATTA 401 ATAAATTGGT AGAAGATTCC GTACTCACTC CTCATAGTAA TACATCACAG 451 ACTAGTCTCA ACAACATCTT CAATAAAAAA TTACACGTCA AAATCGAAAA 501 CAAATCCCAC GTCGCCGGAC AGGTGTTGGA ACTGACCAAG ATGACGCTGA 551 AAGATTCCCT TTGGGAACCG CGCCGCCATT CCGACATCCA TATGCTGGAA 601 ACTTCCGATA ATGCCCGCAT CCGCCTGAAC ACGAAAGATG AAAAACTGAC 651 CGTCCATAAA GCGTATCAGG GCGGTGCGGA TTTCCTGTTC GGCTACGACG 701 TGCGGGAGTC GGACAAACCC GCCCTGACCT TTGAAGAAAA AGTCAGCGGA 751 CAATCCGGCG TGGTTTTGGA ACGCCGGCCG GAAAATCTGA AAACGCTCGA 801 CGGGCGCAAA CTGATTGCGG CGGAAAAGGC AGACTCTAAT TCGTTTGCGT 851 TTAAACAAAA TTACCGGCAG GGACTGTACG AATTATTGCT CAAGCAATGC 901 GAAGGCGGAT TTTGCTTGGG CGTGCAGCGT TTGGCTATCC CCGAGGCGGA 951 AGCGGTTTTA TATGCCCAAC AGGCTTATGC GGCAAATACT TTGTTCGGGC 1001 TGCGTGCCGC CGACAGGGGC GACGACGTGT ATGCCGCCGA TCCGTCCCGT 1051 CAAAAATTGT GGCTGCGCTT CATCGGCGGC CGGTCGCATC AAAATATACG 1101 GGGCGGCGCG GCTGCGGACG GGCGGCGCAA AGGCGTGCAA ATCGGCGGCG 1151 AGGTGTTTGT ACGGCAAAAT GAAGGCAGCC GGCTGGCAAT CGGCGTGATG 1201 GGCGGCAGGG CTGGCCAGCA CGCATCAGTC AACGGCAAAG GCGGTGCGGC 1251 AGGCAGTTAT TTGCATGGTT ATGGCGGGGG TGTTTATGCT GCGTGGCATC 1301 AGTTGCGCGA TAAACAAACG GGTGCGTATT TGGACGGCTG GTTGCAATAC 1351 CAACGTTTCA AACACCGCAT CAATGATGAA AACCGTGCGG AACGCTACAA 1401 AACCAAAGGT TGGACGGCTT CTGTCGAAGG CGGCTACAAC GCGCTTGTGG 1451 CGGAAGGCGT TGTCGGAAAA GGCAATAATG TGCGGTTTTA CCTGCAACCG 1501 CAGGCGCAGT TTACCTACTT GGGCGTAAAC GGCGGCTTTA CCGACAGCGA 1551 GGGGACGGCG GTCGGACTGC TCGGCAGCGG TCAGTGGCAA AGCCGCGCCG 1601 GCATTCGGGC AAAAACCCGT TTTGCTTTGC GTAACGGTGT CAATCTTCAG 1651 CCTTTTGCCG CTTTTAATGT TTTGCACAGG TCAAAATCTT TCGGCGTGGA 1701 AATGGACGGC GAAAAACAGA CGCTGGCAGG CAGGACGGCG CTCGAAGGGC 1751 GGTTCGGCAT TGAAGCCGGT TGGAAAGGCC ATATGTCCGC ACGCATCGGA 1801 TACGGCAAAA GGACGGACGG CGACAAAGAA GCCGCATTGT CGCTCAAATG 1851 GCTGTTTTGA This encodes a protem having amino acid sequence <SEQ ED 454>: 1 MFRAQLGSNT RSTKIGDDAD FSFSDKPKPG TΞHYFSSGKT DQNSSEYGYD 51 EINIQGKNYN SGILAVDNMP WKKYITDTY GDNLKDAVKK QLQDLYKTRP 101 EAWEENKKRT EEAYIEQLGP KFSILKQKNP DLINKLVEDS VLTPHSNTSQ 151 TSLNNIFNKK LHVKIENKSH VAGQVLELTK MTLKDSLWEP RRHSDIHMLE 201 TSDNARIRLN TKDEKLTVHK AYQGGADFLF GYDVRESDKP ALTFEEKVSG 251 QSGWLERRP ENLKTLDGRK LIAAEKADSN SFAFKQNYRQ GLYELLLKQC 301 EGGFCLGVQR LAIPEAEAVL YAQQAYAANT LFGLRAADRG DDVYAADPSR 351 QKLWLRFIGG RSHQNIRGGA AADGRRKGVQ IGGEVFVRQN EGSRLAIGVM 401 GGRAGQHASV NGKGGAAGSY LHGYGGGVYA AWHQLRDKQT GAYLDGWLQY 451 QRFKHRINDE NRAERYKTKG WTASVEGGYN ALVAEGWGK GNNVRFYLQP 501 QAQFTYLGVN GGFTDSEGTA VGLLGSGQWQ SRAGIRAKTR FALRNGVNLQ 551 PFAAFNVLHR SKSFGVEMDG EKQTLAGRTA LEGRFGIEAG WKGHMSARIG 601 YGKRTDGDKE AALSLKWLF* Homology with a predicted ORF from N.sonorrhoeae ORF35 shows 51.7% identity over a 261 aa overlap with a predicted ORF (ORF35ngh) from N gonorrhoeae: orf 35 . pep PCRRQGDDVYAAHASRQKLWLRFIGGRSHQΝIRG 34 : : : I : : I : I I I I I I M : : I orf35ngh FTKVQERDDIAIYAQQAQAAΝTLFALRLΝDKΝSDIFDRTLPRKGLWLRVIDGHSΝQWVQG 370 orf35. pep GAA-ADGWRKGVQIGGEVFVRQNEGSXLAIGVMGGRAGQHASVNGKG—GAAGSDLYGYG 91 : I :: I: I I I I I : I I I I I : III:: I : I I : I I I I::: : : : ::: M I orf35ngh KTAPVEGYRKGVQLGGEVFTWQNESNQLSIGLMGGQAEQRSTFRNPDTDNLTTGNVKGFG 430 orf35.pep GGVYAAWHQLRDKQTGAYLDGWLQYQRFKHRINDENRAERYKTKGWTASVEGGYNALVAE 151 : I I II : I I II : I II II II : I : I : I I II I : I II I I MM Ml I I I : I : I I II I : II orf35ngh AGVYATWHQLQDKQTGAYVDSWMQYQRFRHRINTEYATERFTSKGITASIEAGYNALLAE 490 orf35. pep GIVGKGNNVRFYLQPQAQFTYLGVNGGFTDSEGTAVGLLGSGQWQSRAGIRAKTRFALRN 211 :: MMM I II II II : II I I II I MMM: I : I II I I I II : | :: || :: II : I orf35ngh HFTKKGNSLRVYLQPQAQLTYLGVNGKFSDSENAQVNLLGSRQLQSRVGVQAKAQFAFTN 550 orf35. pep GVNLQPFAAFNVLHRSKSFGVEMDGEKQTLAGRTALEGRFGIEAGWKGHMSA 263 IM: II I: I I :::: I I I I I: I I::::: :: M: I :: I : I I : I :: orf35ngh GVTFQPFVAVNSIYQQKPFGVEIDGDRRVINNKTVIETQLGVAAKIKSHLTLQASFNRQT 610 A partial ORF35ngh nucleotide sequence <SEQ ID 455> is predicted to encode a protein having partial amino acid sequence <SEQ ID 456>: 1 ..KKLRDRNSEY WKEETYHIKS NGRTYPNIPA LFPKHPFDPF ENINNSKKIS 51 FYDKEYTEDY LVGFARGFGV EKRNGEEEKP LRQYFKDCVN TENSNNDNCK 101 ISSFGNYGPI LIKSDIFALA SQIKNSHINS EILSVGNYIE WLRPTLNKLT 151 GWQEHLYAGL DPFHYIEVTD NSHVIGQTID LGALELTNSL WKPRWNSNID 201 YLITKNAEIR FNTKNESLLV KEDYAGGARF RFAYDLKDKV PEIPVLTFEK 251 NITGTSDIIF EGKALDNLKH LDGHQIVKVN DTADKDAFRL SSKYRKGIYT 301 LSLQQRPEGF FTKVQERDDI AIYAQQAQAA NTLFALRLND KNSDIFDRTL 351 PRKGLWLRVI DGHSNQWVQG KTAPVEGYRK GVQLGGEVFT WQNESNQLSI 401 GLMGGQAEQR STFRNPDTDN LTTGNVKGFG AGVYATWHQL QDKQTGAYVD 451 SWMQYQRFRH RINTEYATER FTSKGITASI EAGYNALLAE HFTKKGNSLR 501 VYLQPQAQLT YLGVNGKFSD SENAQVNLLG SRQLQSRVGV QAKAQFAFTN 551 GVTFQPFVAV NSIYQQKPFG VEIDGDRRVI NNKTVIETQL GVAAKIKSHL 601 TLQASFNRQT SKHHHAKQGA LNLQWTF* Based on this prediction, these proteins from Nmeningitidis and N.gonorrhoeae, and their epitopes, could be useful .antigens for vaccines or diagnostics, or for raising antibodies. Example 55 The following partial DNA sequence was identified in N. meningitidis <SEQ ED 457>: 1 .. GCGGAATATG TTCAGTTCTC TATAGATTTG TTCAGTGTGG GTAAATCGGG 51 GGGCGGTATA CCTAAGGCTA AGCCTGTGTT TGATGCGAAA CCGAGATGGG 101 AGGTTGATAG GAAGCTTAAT AAATTGACAA CTCGTGAGCA GGTGGAGAAA 151 AATGTTCAGG AAACGAGAAG AAGGAGTCAG AGTAGTCAGT TTAAAGCCCA 201 TGCGCAACGA GAATGGGAAA ATAAAACAGG GTTAGATTTT AATCATTTTA 251 TAGGTGGTGA TATCAATAAA AAAGGCACAG TAACAGGAGG GCATAGTCTA 301 ACCCGTGGTG ATGTACGGGT GATACAACAA ACCTCGGCAC CTGATAAACA 351 TGGGGT.TTA TCAAGCGACA GTGGAAATTN A This coπesponds to the amino acid sequence <SEQ ID 458; ORF46>: 1 ..AEYVQFSIDL FSVGKSGGGI PKAKPVFDAK PRWEVDRKLN KLTTREQVEK 51 NVQETRRRSQ SSQFKAHAQR EWENKTGLDF NHFIGGDINK KGTVTGGHSL 101 TRGDVRVIQQ TSAPDKHGXL SSDSGNX Further work revealed further partial nucleotide sequence <SEQ ID 459>: 1 ..GCAGTGTGCC TnCCGATGCA TGCACACGCC TCAnATTTGG CAAACGATTC 51 TTTTATCCGG CAGGTTCTCG ACCGTCAGCA TTTCGAACCC GACGGGAAAT 101 ACCACCTATT CGGCAGCAGG GGGGAACTTG CCGAGCGCCA GTCTCATATC 151 GGATTGGGAA AAATACAAAG CCATCAGTTG GGCAACCTGA TGATTCAACA 201 GGCGGCCATT AAAGGAAATA TCGGCTACAT TGTCCGCTTT TCCGATCACG 251 GGCACGAAGT CCATTCCCCs TTCGACAACC ATGCCTCACA TTCCGATTCT 301 GATGAAGCCG GTAGTCCCGT TGACGGATTT AGCCTTTACC GCATCCATTG 351 GGACGGATAC GAACACCATC CCGCCGACGG CTATGACGGG CCACAGGGCG 401 GCGGCTATCC CGCTCCCAAA GGCGCGAGGG ATATATACAG TTACGACATA 451 AAAGGCGTTG CCCAAAATAT CCGCCTCAAC CTGACCGACA ACCGCAGCAC 501 CGGACAACGG CTTGCCGACC GTTTCCACAA TGCCGGTAGT ATGCTGACGC 551 AAGGAGTAGG CGACGGATTC AAACGCGCCA CCCGATACAG CCCCGAGCTG 601 GACAGATCGG GCAATGCCGC CGAAGCCTTC AACGGCACTG CAGATATCGT 651 TAAAAACATC ATCGGCGCTG CAGGAGAAAT TGT This coπesponds to the amino acid sequence <SEQ ID 460; ORF46-l>: 1 ..AVCLPMHAHA SXLANDSFIR QVLDRQHFEP DGKYHLFGSR GELAERQSHI 51 GLGKIQSHQL GNLMIQQAAI KGNIGYIVRF SDHGHEVHSP FDNHASHSDS 101 DEAGSPVDGF SLYRIHWDGY EHHPADGYDG PQGGGYPAPK GARDIYSYDI 151 KGVAQNIRLN LTDNRSTGQR LADRFHNAGS MLTQGVGDGF KRATRYSPEL 201 DRSGNAAEAF NGTADIVKNI IGAAGEI Computer analysis of this amino acid sequence gave the following results: Homology with a predicted ORF from Nsonorrhoeae ORF46 shows 98.2% identity over a 11 laa overlap with a predicted ORF (ORF46ng) from N gonorrhoeae: orf46. pep AEYVQFSIDLFSVGKSGGGIPKAKPVFDAKPRWEVDRKLΝKLTTR 45 II I II I II I I I I I I II II I I I I II I I I I I I orf46ng PKTGVPFDGKGFPΝFEKHVKYDTKLDIQELSGGGIPKAKPVFDAKPRWEVDRKLΝKLTTR 217 orf46. pep EQVEKΝVQETRRRSQSSQFKAHAQREWEΝKTGLDFΝHFIGGDIΝKKGTVTGGHSLTRGDV 105 I I II II II I I I I I II I I II I I I II I I II I I II II I I II I I I I II I I I : II I I II I II I I I orf46ng EQVEKΝVQETRRRSQSSQFKAHAQREWEΝKTGLDFΝHFIGGDIΝKKGAVTGGHSLTRGDV 277 orf46. pep RVIQQTSAPDKHGXLSSDSGΝ 126 II I I II I I II I I I I I II I I I orf46ng RVIQQTSAPDKHGVLSSDSGΝ 298 A partial ORF46ng nucleotide sequence <SEQ ID 461 > is predicted to encode a protein having partial amino acid sequence <SEQ ID 462>: 1 ..RRLKHCCHAR LGSAFHRKQD GAHQRFGRYG ATQRLCRSSH PRLGSPKPQC 51 RTRHRSRQQY LYGSHPHQRD WSCPGKIQLG RHHGTSCRAV ADXRDRICER 101 EIRRQRQXCR CRLGKIPSLS IPKYPLKLEQ RYGKEΝITSS TVPPSΝGKΝV 151 KLADQRHPKT GVPFDGKGFP ΝFEKHVKYDT KLDIQELSGG GIPKAKPVFD 201 AKPRWEVDRK LΝKLTTREQV EKΝVQETRRR SQSSQFKAHA QREWEΝKTGL 251 DFΝHFIGGDI ΝKKGAVTGGH SLTRGDVRVI QQTSAPDKHG VLSSDSGΝ* Further work revealed the complete gonococcal DΝA sequence <SEQ ID 463 >: 1 TTGGGCATTT CCCGCAAAAT ATCCCTTATT CTGTCCATAC TGGCAGTGTG 51 CCTGCCGATG CATGCACACG CCTCAGATTT GGcaAACGAT CCCTTTATCC 101 GgCaggttcT CGaccGTCAG CATTTCGaac ccgacggGAa ATACCaCCTA 151 TTcggCaGCA GGGGGGAGCT TgccnagcGC aacggccATa tcggattggG 201 aaacaTAcaa Agccatcagt tGggccacct gatgattcaa caggcggccg 251 ttgaaggaaA TAtcgGctac attgtccgct tttccgatca cgggcacaaa 301 ttccattcgc ccttcGAcaa ccaTGCCTCA CATTCCGATT CTGACGAAGC 351 CGGTAGTCCC GTTGACGGAT TCAGCCTTTA CCGCATCCAT TGGGACGGAT 401 ACGAACACCA TCCCGCCGAC GGCTATGACG GGCCACAGGG CGGCGGCTAT 451 CCCGCTCCCA AAGGCGCGAG GGATATATAC AGCTACGACA TAAAAGGCGT 501 TGCCCAAAAT ATCCGCCTCA ACCTGACCGA CAACCGCAGC ACCGGACAAC 551 GGCTTGCCGA CCGTTTCCAC AATGCCGGCG CTATGCTGAC GCAAGGAGTA 601 GGCGACGGAT TCAAACGCGC CACCCGATAC AGCCCCGAGC TGGACAGATC 651 GGGCAATGCc gccGAAGCCT TCAACGGCAC TGCAGATATC GTCAAAAACA 701 TCATCGGCGC GGCAGGAGAA ATTGTCGGCG CAGGCGATGC CGTGCagGGT 751 ATAAGCGAAG GCTCAAACAT TGCTGTCATG CACGGCTTGG GTCTGCTTTC 801 CACCGAAAAC AAGATGGCGC GCATCAACGA TTTGGCAGAT ATGGCGCAAC 851 TCAAAGACTA TGCCGCAGCA GCCATCCGCG ATTGGGCAGT CCAAAACCCC 901 AATGCCGCAC AAGGCATAGA AGCCGTCAGC AATATCTTTA TGGCAGCCAT 951 CCCCATCAAA GGGATTGGAG CTGTCCGGGG AAAATACGGC TTGGGCGGCA 1001 TCACGGCACA TCCTGTCAAG CGGTCGCAGA TGGGCGCGAT CGCATTGCCG 1051 AAAGGGAAAT CCGCCGTCAG CGACAATTTT GCCGATGCGG CATACGCCAA 1101 ATACCCGTCC CCTTACCATT CCCGAAATAT CCGTTCAAAC TTGGAGCAGC 1151 GTTACGGCAA AGAAAACATC ACCTCCTCAA CCGTGCCGCC GTCAAACGGC 1201 AAAAATGTCA AACTGGCAGA CCAACGCCAC CCGAAGACAG GCGTACCGTT 1251 TGACGGTAAA GGGTTTCCGA ATTTTGAGAA GCACGTGAAA TATGATACGA 1301 AGCTCGATAT TCAAGAATTA TCGGGGGGCG GTATACCTAA GGCTAAGCCT 1351 GTGTTTGATG CGAAACCGAG ATGGGAGGTT GATAGGAAGC TTAATAAATT 1401 GACAACTCGT GAGCAGGTGG AGAAAAATGT TCAGGAAACG AGAAGAAGGA 1451 GTCAGAGTAG TCAGTTTAAA GCCCATGCGC AACGAGAATG GGAAAATAAA 1501 ACAGGGTTAG ATTTTAATCA TTTTATAGGT GGTGATATCA ATAAGAAAGG 1551 CACAGTAACA GGAGGGCATA GTCTAACCCG TGGTGATGTA CGGGTGATAC 1601 AACAAACCTC GGCACCTGAT AAACATGGGG TTTATCAAGC GACAGTGGAA 1651 ATTAAAAAGC CTGATGGAAG TTGGGAGGTG AAAACGAAAA AAGGTGGGAA 1701 AGTGATGACC AAGCACACCA TGTTCCCAAA AGATTGGGAT GAGGCTAGAA 1751 TTAGGGCTGA AGTTACTTCG GCTTGGGAAA GTAGAATAAT GCTTAAGGAT 1801 AATAAATGGC AGGGTACAAG TAAATCGGGT ATTAAAATAG AAGGATTTAC 1851 CGAACCTAAT AGAACAGCAT ATCCCATTTA TGAATAG This coπesponds to the .amino acid sequence <SEQ ID 464; ORF46ng-l > • 1 LGISRKISLI LSILAVCLPM HAHASDLAND PFIRQVLDRQ HFEPDGKYHL 51 FGSRGELAXR NGHIGLGNIQ SHQLGHLMIQ QAAVEGNIGY IVRFSDHGHK 101 FHSPFDNHAS HSDSDEAGSP VDGFSLYRIH WDGYEHHPAD GYDGPQGGGY 151 PAPKGARDIY SYDIKGVAQN IRLNLTDNRS TGQRLADRFH NAGAMLTQGV 201 GDGFKRATRY SPELDRSGNA AEAFNGTADI VKNIIGAAGE IVGAGDAVQG 251 ISEGSNIAVM HGLGLLSTEN KMARINDLAD MAQLKDYAAA AIRDWAVQNP 301 NAAQGIEAVS NIFMAAIPIK GIGAVRGKYG LGGITAHPVK RSQMGAIALP 351 KGKSAVSDNF ADAAYAKYPS PYHSRNIRSN LEQRYGKENI TSSTVPPSNG 401 KNVKLADQRH PKTGVPFDGK GFPNFEKHVK YDTKLDIQEL SGGGIPKAKP 451 VFDAKPRWEV DRKLNKLTTR EQVEKNVQET RRRSQSSQFK AHAQREWENK 501 TGLDFNHFIG GDINKKGTVT GGHSLTRGDV RVIQQTSAPD KHGVYQATVE 551 IKKPDGSWEV KTKKGGKVMT KHTMFPKDWD EARIRAEVTS AWESRIMLKD 601 NKWQGTSKSG IKIEGFTEPN RTAYPIYE* ORF46ng-l and ORF46-1 show 94.7% identity in 227 aa overlap: 10 20 30 40 orf46-1. pep AVCLPMHAHASXLANDSFIRQVLDRQHFEPDGKYHLFGSRGELAER I II I II I I II I I II I II I I I I I II I I I I I I I I I II I I I I I II I orf46ng-l LGISRKISLILSILAVCLPMHAHASDLANDPFIRQVLDRQHFEPDGKYHLFGSRGELAXR 10 20 30 40 50 60 50 60 70 80 90 100 orf46-1. pep QSHIGLGKIQSHQLGNLMIQQAAIKGNIGYIVRFSDHGHEVHSPFDNHASHSDSDEAGSP :: II II I : I I I I I I I : I II I I II :: I I I I I I I I II II I I : I I I I II I I I I I I I II I I I I orf46ng-l NGHIGLGNIQSHQLGHLMIQQAAVEGNIGYIVRFSDHGHKFHSPFDNHASHSDSDEAGSP 70 80 90 100 110 120 110 120 130 140 150 160 orf46-1. pep VDGFSLYRIHWDGYEHHPADGYDGPQGGGYPAPKGARDIYSYDIKGVAQNIRLNLTDNRS I I I I I I I I II II I II I I I I II I I I I II I I II I I I I I I I I I I I I I I I I I II II I I I I I II I orf46ng-l VDGFSLYRIHWDGYEHHPADGYDGPQGGGYPAPKGARDIYSYDIKGVAQNIRLNLTDNRS 130 140 150 160 170 180 170 180 190 200 210 220 orf46-1. pep TGQRLADRFHNAGSMLTQGVGDGFKRATRYSPELDRSGNAAEAFNGTADIVKNIIGAAGE I I I II I II II I IM I I I I I I I I I I II II I I II I II II II I I I I I I II I I I I I I II II I I I orf46ng-l TGQRLADRFHNAGAMLTQGVGDGFKRATRYSPELDRSGNAAEAFNGTADIVKNIIGAAGE 190 200 210 220 230 240 orf46-l.pep I I orf46ng-l IVGAGDAVQGISEGSNIAVMHGLGLLSTENKMARINDLADMAQLKDYAAAAIRDWAVQNP 250 260 270 280 290 300 Homology with a predicted ORF from N. meningitidis (strain A) ORF46ng-l shows 87.4% identity over a 486aa overlap with an ORF (ORF46a) from strain A of N. meningitidis: 10 20 30 40 50 60 orf46a.pep LGISRKISLILSILAVCLPMHAHASDLANDSFIRQVLDRQHFEPDGKYHLFGSRGELAER I II I II I I I I II II I II I II I I I I I I I II I II I I II II I I II II I I I I II II I I II I I orf46ng-l LGISRKISLILSILAVCLPMHAHASDLANDPFIRQVLDRQHFEPDGKYHLFGSRGELAXR 10 20 30 40 50 60 70 80 90 100 110 120 orf46a . pep SGHIGLGNIQSHQLGNLFIQQAAIKGNIGYIVRFSDHGHEVHSPFDNHASHSDSDEAGSP : II I I II II I I I I II : I : II I I I :: I I I I I I I I II II II : II II I I I I I I II I I I II II orf46ng-l NGHIGLGNIQSHQLGHLMIQQAAVEGNIGYIVRFSDHGHKFHSPFDNHASHSDSDEAGSP 70 80 90 100 110 120 130 140 150 160 170 180 orf46a . pep VDGFSLYRIHWDGYEHHPADGYDGPQGGGYPAPKGARDIYSYDIKGVAQNIRLNLTDNRS I I I II II II II I I II II I II I I I I I II I I I I II I II I I I II I I I I I I I I I II I II II I I I orf46ng-l VDGFΞLYRIHWDGYEHHPADGYDGPQGGGYPAPKGARDIYSYDIKGVAQNIRLNLTDNRS 130 140 150 160 170 180 190 200 210 220 230 240 orf46a . pep TGQRLVDRFHNTGSMLTQGVGDGFKRATRYSPELDRSGNAAEAFNGTADIVKNIIGAAGE I I I I I : I I II I : I : I I I II I I II I I I I I I I I I I I I II II II I I I I I I II I II I I II II II orf46ng-l TGQRLADRFHNAGAMLTQGVGDGFKRATRYSPELDRSGNAAEAFNGTADIVKNIIGAAGE 190 200 210 220 230 240 250 260 270 280 290 300 orf46a . pep IVGAGDAVQGISEGSNIAVMHGLGLLSTENKMARINDLADMAQLKDYAAAAIRDWAVQNP I II I I I I I I I II II II I II I I II I II I I I I I II I II I I I II I I I I I I I I I I I I I I I II I I orf46ng-l IVGAGDAVQGISEGSNIAVMHGLGLLSTENKMARINDLADMAQLKDYAAAAIRDWAVQNP 250 260 270 280 290 300 310 320 330 340 350 360 orf46a. pep NAAQGIEAVSNIFTAVIPVKGIGAVRGKYGLGGITAHPVKRSQMGEIALPKGKSAVSDNF I I I I I II II II I I I : II : I I I I I II I I I I I I I I II I II II I I I I I I I I II I I I II I I I orf46ng-l NAAQGIEAVSNIFMAAIPIKGIGAVRGKYGLGGITAHPVKRSQMGAIALPKGKSAVSDNF 310 320 330 340 350 360 370 380 390 400 410 420 orf46a . pep ADAAYAKYPSPYHSRNIRSNLEQRYGKENITSSTVPPSNGKNVKLANKRHPKTKVPFDGK I I I II II II II I II II II I I I II I II I I I I I I II I I II II I I I I II :: I I I II I I II I I orf46ng-l ADAAYAKYPSPYHSRNIRSNLEQRYGKENITSSTVPPSNGKNVKLADQRHPKTGVPFDGK 370 380 390 400 410 420 430 440 450 460 470 orf46a . pep GFPNFEKDVKYDTRINTAVPQVN PIDEPVFN—PKGSVGSAHSWSITARIQYAKLP I I I II I I I I II I :: : : ::: I : I I I : I : I : : : I : I I I orf46ng-l GFPNFEKHVKYDTKLD--IQELSGGGIPKAKPVFDAKPRWEVDRKLN-KLTTREQVEKNV 430 440 450 460 470 480 490 500 510 520 530 orf46a . pep RQGRIRYIPPKNYSPSAPLPKGPNNGYLDKFGNEWTKGPSRTKGQEFEWDVQLSKTGREQ :: I I orf46ng-l QETRRRSQSSQFKAHAQREWENKTGLDFNHFIGGDINKKGTVTGGHSLTRGDVRVIQQTS 480 490 500 510 520 530 The complete length ORF46a DNA sequence <SEQ ID 465> is: 1 TTGGGCATTT CCCGCAAAAT ATCCCTTATT CTGTCCATAC TGGCAGTGTG 51 CCTGCCGATG CATGCACACG CCTCAGATTT GGCAAACGAT TCTTTTATCC 101 GGCAGGTTCT CGACCGTCAG CATTTCGAAC CCGACGGGAA ATACCACCTA 151 TTCGGCAGCA GGGGGGAACT TGCCGAGCGC AGCGGTCATA TCGGATTGGG 201 AAACATACAA AGCCATCAGT TGGGCAACCT GTTCATCCAG CAGGCGGCCA 251 TTAAAGGAAA TATCGGCTAC ATTGTCCGCT TTTCCGATCA CGGGCACGAA 301 GTCCATTCCC CCTTCGACAA CCATGCCTCA CATTCCGATT CTGATGAAGC 351 CGGTAGTCCC GTTGACGGAT TCAGCCTTTA CCGCATCCAT TGGGACGGAT 401 ACGAACACCA TCCCGCCGAC GGCTATGACG GGCCACAGGG CGGCGGCTAT 451 CCCGCTCCCA AAGGCGCGAG GGATATATAC AGCTACGACA TAAAAGGCGT 501 TGCCCAAAAT ATCCGCCTCA ACCTGACCGA CAACCGCAGC ACCGGACAAC 551 GGCTTGTCGA CCGTTTCCAC AATACCGGTA GTATGCTGAC GCAAGGAGTA 601 GGCGACGGAT TCAAACGCGC CACCCGATAC AGCCCCGAGC TGGACAGATC 651 GGGCAATGCC GCCGAAGCTT TCAACGGCAC TGCAGATATC GTCAAAAACA 701 TCATCGGCGC GGCAGGAGAA ATTGTCGGCG CAGGCGATGC CGTGCAGGGT 751 ATAAGCGAAG GCTCAAACAT TGCTGTTATG CACGGCTTGG GTCTGCTTTC 801 CACCGAAAAC AAGATGGCGC GCATCAACGA TTTGGCAGAT ATGGCGCAAC 851 TCAAAGACTA TGCCGCAGCA GCCATCCGCG ATTGGGCAGT CCAAAACCCC 901 AATGCCGCAC AAGGCATAGA AGCCGTCAGC AATATCTTTA CGGCAGTCAT 951 CCCCGTCAAA GGGATTGGAG CTGTTCGGGG AAAATACGGC TTGGGCGGCA 1001 TCACGGCACA TCCTGTCAAG CGGTCGCAGA TGGGCGAGAT CGCATTGCCG 1051 AAAGGGAAAT CCGCCGTCAG CGACAATTTT GCCGATGCGG CATACGCCAA 1101 ATACCCGTCC CCTTACCATT CCCGAAATAT CCGTTCAAAC TTGGAGCAGC 1151 GTTACGGCAA AGAAAACATC ACCTCCTCAA CCGTGCCGCC GTCAAACGGA 1201 AAGAATGTGA AACTGGCAAA CAAACGCCAC CCGAAGACCA AAGTGCCGTT 1251 TGACGGTAAA GGGTTTCCGA ATTTTGAAAA AGACGTAAAA TACGATACGA 1301 GAATTAATAC CGCTGTACCA CAAGTGAATC CTATAGATGA ACCCGTCTTT 1351 AATCCTAAAG GTTCTGTCGG ATCGGCTCAT TCTTGGTCTA TAACTGCCAG 1401 AATTCAATAC GCAAAATTAC CAAGGCAAGG TAGAATCAGA TATATCCCAC 1451 CTAAAAATTA CTCTCCTTCA GCACCGCTAC CAAAAGGACC TAATAATGGA 1501 TATTTGGATA AATTTGGTAA TGAATGGACT AAAGGTCCAT CAAGAACTAA 1551 AGGTCAAGAA TTTGAATGGG ATGTTCAATT GTCTAAAACA GGAAGAGAGC 1601 AACTTGGATG GGCTAGTAGG GATGGTAAGC ATTTAAATAT ATCAATTGAT 1651 GGAAAGATTA CACACAAATG A This coπesponds to the amino acid sequence <SEQ ID 466>: 1 LGISRKISLI LSILAVCLPM HAHASDLAND SFIRQVLDRQ HFEPDGKYHL 51 FGSRGELAER SGHIGLGNIQ SHQLGNLFIQ QAAIKGNIGY IVRFSDHGHE 101 VHSPFDNHAS HSDSDEAGSP VDGFSLYRIH WDGYEHHPAD GYDGPQGGGY 151 PAPKGARDIY SYDIKGVAQN IRLNLTDNRS TGQRLVDRFH NTGSMLTQGV 201 GDGFKRATRY SPELDRSGNA AEAFNGTADI VKNIIGAAGE IVGAGDAVQG 251 ISEGSNIAVM HGLGLLSTEN KMARINDLAD MAQLKDYAAA AIRDWAVQNP 301 NAAQGIEAVS NIFTAVIPVK GIGAVRGKYG LGGITAHPVK RSQMGEIALP 351 KGKSAVSDNF ADAAYAKYPS PYHSRNIRSN LEQRYGKENI TSSTVPPSNG 401 KNVKLANKRH PKTKVPFDGK GFPNFEKDVK YDTRINTAVP QVNPIDEPVF 451 NPKGSVGSAH SWSITARIQY AKLPRQGRIR YIPPKNYSPS APLPKGPNNG 501 YLDKFGNEWT KGPSRTKGQE FEWDVQLSKT GREQLGWASR DGKHLNISID 551 GKITHK* B.ased on this analysis, including the presence of a RGD sequence in the gonococcal protein, typical of adhesins, it is predicted that the proteins from N. meningitidis and N.gonorrhoeae, and their epitopes, could be useful antigens for vaccines or diagnostics, or for raising antibodies. Example 56 The following partial DNA sequence was identified in N. meningitidis <SEQ ID 467>: 1 ATGAATATTC ACACCCTGCT CTCCAAACAA TGGACGCTGC CGCCATTCCT 51 GCCGAAACGG CTGCTGCTGT CCCTGCTGAT ACTGCTTGCC CCCAATGCGG 101 TGTTTTGGGT TTTGGCACTG CTGACCGCCA CCGCCCGCCC GATTGTCAAT 151 TTGGACTATC TTCCCGCCGC GCTGCTGATC GCCCTGCCTT GGCGTTTCGT 201 CAAAATTGCC GGCGTATTGG CGTTTTGGCT GGCGGTTTTG TTTGACGGGC 251 TGATGATGGT GATCCAACTC TTCCCTTTTA TGGATCTCAT CGGCGCCATC 301 AACCTCGTCC CCTTCATCCT GACCGCCCCC GCCCCTTATC AGATAATGAC 351 CGGGCTG... This coπesponds to the amino acid sequence <SEQ ID 468; ORF48>: 1 MNIHTLLSKQ WTLPPFLPKR LLLSLLILLA PNAVFWVLAL LTATARPIVN 51 LDYLPAALLI ALPWRFVKIA GVLAFWLAVL FDGLMMVIQL FPFMDLIGAI 101 NLVPFILTAP APYQIMTGL... Further work revealed the complete nucleotide sequence <SEQ LD 469>: 1 ATGAATATTC ACACCCTGCT CTCCAAACAA TGGACGCTGC CGCCATTCCT 51 GCCGAAACGG CTGCTGCTGT CCCTGCTGAT ACTGCTTGCC CCCAATGCGG 101 TGTTTTGGGT TTTGGCACTG CTGACCGCCA CCGCCCGCCC GATTGTCAAT 151 TTGGACTATC TTCCCGCCGC GCTGCTGATC GCCCTGCCTT GGCGTTTCGT 201 CAAAATTGCC GGCGTATTGG CGTTTTGGCT GGCGGTTTTG TTTGACGGGC 251 TGATGATGGT GATCCAACTC TTCCCTTTTA TGGATCTCAT CGGCGCCATC 301 AACCTCGTCC CCTTCATCCT GACCGCCCCC GCCCCTTATC AGATAATGAC 351 CGGGCTGTTG CTGCTGTATA TGCTGGCGAT GCCGTTTGTG TTGCAGAAAG 401 CCGCCGCCAA AACCGACTTC CGGCACATTG CCGTCTGCGC CGCCGTTGTG 451 GCGGCAGCCG GCTATTTCAC CGGCCATTTG AGTTACTACG ACCGGGGTCG 501 GATGGCCAAT ATCTTCGGCG CAAACAACTT CTACTACGCC AAAAGTCAGG 551 CGATGCTCTA CACCGTCAGC CAGAATGCCG ACTTTATTAC CGCCGGCCTG 601 GTCGATCCCG TCTTCCTCCC CTTGGGCAAT CAACAGCGTG CCGCCACGCA 651 TCTGAACGAG CCGAAATCTC AAAAAATCCT CTTTATCGTC GCCGAATCTT 701 GGGGGCTGCC GGCCAATCCC GAACTTCAAA ACGCCACTTT TGCCAAACTG 751 CTGGCGCAAA AAGACCGTTT TTCGGTTTGG GAAAGCGGCA GTTTTCCCTT 801 CATCGGCGCG ACGGTCGAAG GCGAAATGCG CGAACTGTGT GCCTACGGCG 851 GTTTGCGCGG GTTCGCACTG CGCCGCGCGC CCGACGAAAA ATTTGCCCGC 901 TGCCTCCCCA ACCGTTTGAA ACAAGAAGGT TACGCCACCT TTGCGATGCA 951 CGGCGCGGGC AGTTCGCTTT ACGACCGCTT CAGCTGGTAT CCGAGGGCGG 1001 GCTTTCAAGA AATCAAAACC GCCGAAAACC TGATCGGTAA AAAAACCTGC 1051 GCCATTTTCG GCGGCGTGTG CGACAGCGAG CTGTTCGGCG AAGTGTCGGC 1101 ATTTTTCAAA AAACACGACA AGGGACTGTT TTACTGGATG ACGCTGACCA 1151 GCCACGCCGA CTATCCCGAA TCCGACATTT TCAACCACAG GCTCAAATGC 1201 ACCGAATATG GCCTGCCCGC CGAAACCGAC CTCTGCCGCA ATTTCAGCCT 1251 GCACACCCAA TTCTTCGACC AACTGGCGGA TTTGATCCAA CGCCCCGAAA 1301 TGAAAGGCAC GGAAGTCATC ATCGTCGGCG ACCATCCGCC GCCCGTCGGC 1351 AACCTCAATG AAACCTTCCG CTACCTCAAA CAGGGGCACG TCGCCTGGCT 1401 GAACTTCAAA ATCAAATAA This coπesponds to the amino acid sequence <SEQ ID 470; ORF48-l>: 1 MNIHTLLSKQ WTLPPFLPKR LLLSLLILLA PNAVFWVLAL LTATARPIVN 51 LDYLPAALLI ALPWRFVKIA GVLAFWLAVL FDGLMMVIQL FPFMDLIGAI 101 NLVPFILTAP APYQIMTGLL LLYMLAMPFV LQKAAAKTDF RHIAVCAAW 151 AAAGYFTGHL SYYDRGRMAN IFGANNFYYA KSQAMLYTVS QNADFITAGL 201 VDPVFLPLGN QQRAATHLNE PKSQKILFIV AESWGLPANP ELQNATFAKL 251 LAQKDRFSVW ESGSFPFIGA TVEGEMRELC AYGGLRGFAL RRAPDEKFAR 301 CLPNRLKQEG YATFAMHGAG SSLYDRFSWY PRAGFQEIKT AENLIGKKTC 351 AIFGGVCDSE LFGEVSAFFK KHDKGLFYWM TLTSHADYPE SDIFNHRLKC 401 TEYGLPAETD LCRNFSLHTQ FFDQLADLIQ RPEMKGTEVI IVGDHPPPVG 451 NLNETFRYLK QGHVAWLNFK IK* Computer analysis of this amino acid sequence gave the following results: Homology with a predicted ORF from N.meninsitidis (strain Al ORF48 shows 94.1% identity over a 119aa overlap with an ORF (ORF48a) from strain A of N. meningitidis: 10 20 30 40 50 60 orf48. pep MNIHTLLSKQWTLPPFLPKRLLLSLLILLAPNAVFWVLALLTATARPIVNLDYLPAALLI I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I II I I I I I I I I II I II I I I I I I I II I I I orf48a MNIHTLLSKQWTLPPFLPKRLLLSLLILLXPNAVFWVLALLTATARPIVNLXYLPAALLI 10 20 30 40 50 60 70 80 90 100 110 119 orf48. pep ALPWRFVKIAGVLAFWLAVLFDGLMMVIQLFPFMDLIGAINLVPFILTAPAPYQIMTGL I I I II III II II I I I II I II II I I I I I I II I I II II I I I I I II I I I I II II I I I orf48a ALPWRXVKIXGVLAXWLAVLFDGLMMVIQLFPFMDLIGAINLVPFIXTAPALYQIMTGLL 70 80 90 100 110 120 orf48a LLYMLAMPFVLQKAAAKTDFRHIAACAAVWAAGYFTGHLSXYDRGRMANIFGANNFYYA 130 140 150 160 170 180 The complete length ORF48a nucleotide sequence <SEQ D 471> is: 1 ATGAATATTC ACACCCTGCT CTCCAAACAA TGGACGCTGC CGCCATTCCT 51 GCCGAAACGG CTGCTGCTGT CCCTGCTGAT ACTGCTNNCC CCCAATGCGG 101 TGTTTTGGGT TTTGGCACTG CTGACCGCCA CCGCCCGCCC GATTGTCAAT 151 TTGGANTACC TTCCCGCCGC GCTGCTGATC GCCCTGCCTT GGCGTNTCGT 201 CAAAATTGNC GGCGTATTGG CGTNTTGGCT GGCGGTTTTG TTTGACGGGC 251 TGATGATGGT GATCCAACTC TTCCCTTTTA TGGATCTCAT CGGCGCCATC 301 AACCTCGTCC CCTTCATCNT GACCGCCCCC GCCCTTTATC AGATAATGAC 351 CGGGCTGTTA CTGCTGTATA TGCTGGCGAT GCCGTTTGTG TTGCAGAAAG 401 CCGCCGCCAA AACCGACTTC CGACACATTG CCGCCTGTGC CGCCGTTGTG 451 GTGGCAGCCG GCTATTTTAC CGGCCATTTG AGTTANTACG ACCGGGGGCG 501 GATGGCCAAT ATCTTCGGCG CAAACAACTT CTATTACGCC AAAAGTCAGG 551 CGATGCTCTA CACCGTCAGC CAGAATGCCG ACTTTATTAC CGCCGGCCTG 601 GTCGATCCCG TCTTCCTCCC CTTGGGCAAT CAACAGCGTG CCGCCACGCA 651 TCTGAACGAG CCGAAATCTC AAAAAATCCT CTTTATCGTC GCCGAATCTT 701 GGGGGCTGCC GGCCAATCCC GAACTTCAAA ACGCCACTTT TGCCAAACTG 751 CTGGCGCAAA AAGANCGTTT TTCGGTTTGG GAAAGCGGCA GTTTTCCCTT 801 CATCGGCGCG ACGATCGAAG GCGAAATGCG CGAACTGTGT GCCTACGGCG 851 GTTTGCGCGG GTTCGCACTG CGCCGCGCGC CCGACGAAAA ATTTGCCCGC 901 TGCCTCCCCA ACCGTTTGAA ACAAGAAGGT TACGCCACCT TTGCGATGCA 9 95511 CGGCGCGGGC AGTTCGCTTT ACGACCGCTT CAGCTGGTAT CCGAGGGCGG 1001 GCTTTCAAGA AATCAAAACC GCCGAAAACC TGATCGGTAA AAAAACCTGC 1051 GCCATTTTCG GCGGCGTGTG CGACAGCGAG CTGTTCGGCG AAGTGTCGGC 1101 ANTTTTCAAA AAACACGACA AGGGACTGTT TTACTGGATG ACGCTGACCA 1151 GCCACGCCGA CTATCCCGAA TCNGACATTT TCAACCACAG GCTCAAATGC 1 1220011 ACCGAATATG GCCTGCCCGC CGAAACCGAC NTCTGCCGCA ATTTCAGCCT 1251 GCACACCCAA TTCTTCGACC AACTGGCGGA TTTGATCCAA CGCCCCGAAA 1301 TGAAAGGCAC GGAAGTCATC ATCGTCGGCG ACCATCCGCC GCCCGTCGGC 1351 AACCTCAATG AAACCTTCCG CTACCTCAAA CAGGGGCACG TCGNCTGGCT 1401 GAACTTCAAA ATCAAATAA This encodes a protein having amino acid sequence <SEQ LD 472>: 1 MNIHTLLSKQ WTLPPFLPKR LLLSLLILLX PNAVFWVLAL LTATARPIVN 51 LXYLPAALLI ALPWRXVKIX GVLAXWLAVL FDGLMMVIQL FPFMDLIGAI 101 NLVPFIXTAP ALYQIMTGLL LLYMLAMPFV LQKAAAKTDF RHIAACAAW 151 VAAGYFTGHL SXYDRGRMAN IFGANNFYYA KSQAMLYTVS QNADFITAGL 201 VDPVFLPLGN QQRAATHLNE PKSQKILFIV AESWGLPANP ELQNATFAKL 251 LAQKXRFSVW ESGSFPFIGA TIEGEMRELC AYGGLRGFAL RRAPDEKFAR 301 CLPNRLKQEG YATFAMHGAG SSLYDRFSWY PRAGFQEIKT AENLIGKKTC 351 AIFGGVCDSE LFGEVSAXFK KHDKGLFYWM TLTSHADYPE SDIFNHRLKC 401 TEYGLPAETD XCRNFSLHTQ FFDQLADLIQ RPEMKGTEVI IVGDHPPPVG 451 NLNETFRYLK QGHVXWLNFK IK* ORF48a and ORF48-1 show 96.8% identity in 472 aa overlap: 10 20 30 40 50 60 orf48a. pep MNIHTLLSKQWTLPPFLPKRLLLSLLILLXPNAVFWVLALLTATARPIVNLXYLPAALLI I I II I I II I II I I I II I II I II II I I I I I I I I II I I I I I II II II I I I I I II I I I I II orf48-1 MNIHTLLSKQWTLPPFLPKRLLLSLLILLAPNAVFWVLALLTATARPIVNLDYLPAALLI 10 20 30 40 50 60 70 80 90 100 110 120 orf48a. pep ALPWRXVKIXGVLAXWLAVLFDGLMMVIQLFPFMDLIGAINLVPFIXTAPALYQIMTGLL I II I I III I I I I I II II I II I II I II I I I I I I I I I I I I I I I I I II I I I I I I I I I I orf48-1 ALPWRFVKIAGVLAFWLAVLFDGLMMVIQLFPFMDLIGAINLVPFILTAPAPYQIMTGLL 70 80 90 100 110 120 130 140 150 160 170 180 orf48a. pep LLYMLAMPFVLQKAAAKTDFRHIAACAAVWAAGYFTGHLSXYDRGRMANIFGANNFYYA II I I I I I I I I II I II I I II I I I I I : I I I I I : I II I I I I I I I II II I I II I I II I I I I I I orf48-1 LLYMLAMPFVLQKAAAKTDFRHIAVCAAWAAAGYFTGHLSYYDRGRMANIFGANNFYYA 130 140 150 160 170 180 190 200 210 220 230 240 orf48a. pep KSQAMLYTVSQNADFITAGLVDPVFLPLGNQQRAATHLNEPKSQKILFIVAESWGLPANP I I I I I I II I I I I I I I I I I II I I I I I I I I I I II I I I II I II I II I I I II I I II I I I I I I I I orf48-1 KSQAMLYTVSQNADFITAGLVDPVFLPLGNQQRAATHLNEPKSQKILFIVAESWGLPANP 190 200 210 220 230 240 250 260 270 280 290 300 orf48a. pep ELQNATFAKLLAQKXRFSVWESGSFPFIGATIEGEMRELCAYGGLRGFALRRAPDEKFAR I I I I I II I I I II I I II II I I I I I I I I I I I I: I II I I I I I I I I II I I I I I I I I II I I I I I orf48-1 ELQNATFAKLLAQKDRFSVWESGSFPFIGATVEGEMRELCAYGGLRGFALRRAPDEKFAR 250 260 270 280 290 300 310 320 330 340 350 360 orf48a. pep CLPNRLKQEGYATFAMHGAGSSLYDRFSWYPRAGFQEIKTAENLIGKKTCAIFGGVCDSE I I I II I I I II I I I I I I II II I I I I I I I I II I II I I I II I I I II I I I II I I II I I I I I I I I orf48-1 CLPNRLKQEGYATFAMHGAGSSLYDRFSWYPRAGFQEIKTAENLIGKKTCAIFGGVCDSE 310 320 330 340 350 360 370 380 390 400 410 420 orf48a. pep LFGEVSAXFKKHDKGLFYWMTLTSHADYPESDIFNHRLKCTEYGLPAETDXCRNFSLHTQ I I I II I I I I I I I I I I I I I I II I II I II I I I I I I I I II II I I I I II I I I I I II I I I I I I orf48-1 LFGEVSAFFKKHDKGLFYWMTLTSHADYPESDIFNHRLKCTEYGLPAETDLCRNFSLHTQ 370 380 390 400 410 420 430 440 450 460 470 orf48a. pep FFDQLADLIQRPEMKGTEVIIVGDHPPPVGNLNETFRYLKQGHVXWLNFKIKX II I I I I II II I I I II II II II I I I I II II II I I II I I I I II I II I I I I I II I orf 8-1 FFDQLADLIQRPEMKGTEVIIVGDHPPPVGNLNETFRYLKQGHVAWLNFKIKX 430 440 450 460 470 Homology with a predicted ORF from N.sonorrhoeae ORF48 shows 97.5% identity over a 119aa overlap with a predicted ORF (ORF48ng) from N gonorrhoeae: orf48.pep MΝIHTLLSKQWTLPPFLPKRLLLSLLILLAPΝAVFWVLALLTATARPIVΝLDYLPAALLI 60 I I I I: II I : I I I II I I I I I I I I I I I II I I II II II I I I I I I II I I I I I II I I I I I II I I I orf 8ng MΝIHALLSEQWTLPPFLPKRLLLSLLILLAPΝAVFWVLALLTATARPIVΝLDYLPAALLI 60 orf48. pep ALPWRFVKIAGVLAFWLAVLFDGLMMVIQLFPFMDLIGAIΝLVPFILTAPAPYQIMTGL 119 I I I II I I I I I II I II I II II I I II I II I I I I II I I I II I I I I I I II I I I II II I I I I I orf48ng ALPWRFVKIAGVLAFWPAVLFDGLMMVIQLFPFMDLIGAIΝLVPFILTAPAPYQIMTGLL 120 The ORF48ng nucleotide sequence <SEQ LD 473> was predicted to encode aprotein having amino acid sequence <SEQ LD 474>: 1 MΝIHALLSEQ WTLPPFLPKR LLLSLLILLA PΝAVFWVLAL LTATARPIVΝ 51 LDYLPAALLI ALPWRFVKIA GVLAFWPAVL FDGLMMVIQL FPFMDLIGAI 101 ΝLVPFILTAP APYQIMTGLL LLYMLAMPFV LQKAAVKTDF RHIAVCAAW 151 AAARYFTGPF ELLRTGGRWQ YVQHRRLLLS GSRASFRRRQ KADVLRRLGΝ 201 PYASMGΝGG.. Further work identified the complete gonococcal DΝA sequence <SEQ ID 475>: 1 ATGAATATTC ACGCCCTGCT CTCCGAACAA TGGACGCTGC CGCCATTCCT 51 GCCGAAACGG CTGCTGCTGT CCCTGCTGAT ACTGCTGGCC CCCAATGCGG 101 TGTTTTGGGT TTTGGCACTG CTGACCGCCA CCGCCCGCCC GATTGTCAAT 151 TTGGACTACC TTCCCGCCGC GCTGCTGATC GCCCTGCCTT GGCGTTTCGT 201 CAAAATTGCC GGCGTATTGG CGTTTTGGCC GGCGGTTTTG TTTGACGGGC 251 TGATGATGGT GATCCAACTC TTCCCTTTTA TGGACCTCAT CGGCGCCATC 301 AACCTCGTCC CCTTCATCCT GACCGCCCCC GCCCCTTATC AGATAATGAC 351 CGGGCTGTTG CTGCTGTATA TGCTGGCGAT GCCGTTTGTG TTGCAAAAAG 401 CCGCCGTCAA AACCGACTTC CGACACATTG CCGTCTGTGC CGCCGTTGTG 451 GCGGCAGCCG GCTATTTCAC CGGCCATTTG AGTTACTACG ACCGGGGGCG 501 GATGGCCAAT ATCTTCGGCG CAAACAACTT CTATTACGCc aAAAGTCAGG 551 CGATGCTCTA CACCGTCAGC CAGAATGCCG ACTTTATTAC CGCCGgcctG 601 GTCGACCCCG TCTTCCTCCC CTTGGGCAAT CAGCAGCGTG CCGCCACGCG 651 GCTGAGTGAG CCGAAATCTC AAAAAATCCT CTTTATCGTC GCCGAATCTT 701 GGGGGCTGCC GGGCAATCCC GAGCTTCAAA ACGCCACTTT TGCCAAACTG 751 CTGGCGCAAA AAGACCGTTT TTCGGTTTGG GAAAGCGGCA GTTTTCCCTT 801 CATCGGCGCG ACGGTCGAAG GCGAAATGCG CGAATTGTGC GCCTACGGCG 851 GTTTGCGCGG GTTCGCACTG CGCCGCGCGC CCGACGAAAA ATTTGCCCGC 901 TGCCTCCCCA ACCGTTTGAA ACAAGAAGGT TACGCCACCT TTGCGATGCA 951 CGGCGCGGGT AGTTCGCTTT ACGACCGCTT CAGCTGGTAT CCGAGGGCGG 1001 GCTTTCAAAA AATCAAAACC GCCGAAAACC TGATCGGTAA AAAAACCTGC 1051 GCCATTTTCG GCGGCGTGTG CGACAGCGAG CTGTTCGGCG AAGTGTCGGC 1101 ATTTTTCAAA AAACACGACA AGGGACTGTT TTACTGGATG ACGCTGACCA 1151 GCCACGCCGA CTATCCCGAA TCCGACATTT TCAACCACAG GCTCAAATGC 1201 ACCGAATACG GCCTGCCCGC CGAAACCGAC CTCTGCCGCA ATTTCAGCCT 1251 GCACACCCAA TtcttcgACC AACTGGCGGA TTTGATCCGA CGCCCCGAAA 1301 TGAAAGGCAC GGAAGTCATC ATCGTCGGCG ACCATCCGCC GCCCGTCGGC 1351 AACCTCAATG AAACCTTCCG CTACCTCAAA CAGGGACACG TCGCCTGGCT 1401 GCACTTCAAA ATCAAATAA This encodes a protein having amino acid sequence <SEQ LD 476; ORF48ng-l>: 1 MNIHALLSEQ WTLPPFLPKR LLLSLLILLA PNAVFWVLAL LTATARPIVN 51 LDYLPAALLI ALPWRFVKIA GVLAFWPAVL FDGLMMVIQL FPFMDLIGAI 101 NLVPFILTAP APYQIMTGLL LLYMLAMPFV LQKAAVKTDF RHIAVCAAW 151 AAAGYFTGHL SYYDRGRMAN IFGANNFYYA KSQAMLYTVS QNADFITAGL 201 VDPVFLPLGN QQRAATRLSE PKSQKILFIV AESWGLPGNP ELQNATFAKL 251 LAQKDRFSVW ESGSFPFIGA TVEGEMRELC AYGGLRGFAL RRAPDEKFAR 301 CLPNRLKQEG YATFAMHGAG SSLYDRFSWY PRAGFQKIKT AENLIGKKTC 351 AIFGGVCDSE LFGEVSAFFK KHDKGLFYWM TLTSHADYPE SDIFNHRLKC 401 TEYGLPAETD LCRNFSLHTQ FFDQLADLIR RPEMKGTEVI IVGDHPPPVG 451 NLNETFRYLK QGHVAWLHFK IK* ORG48ng-l and ORF48-1 show 97.9% identity in 472 aa overlap: 10 20 30 40 50 60 orf48-1. pep MNIHTLLSKQWTLPPFLPKRLLLSLLILLAPNAVFWVLALLTATARPIVNLDYLPAALLI I I I I: I I I : II II I I I I I I I I I I I II I I I II I I I I I II II I I I I I I I I I I II I II I I II I orf48ng-l MNIHALLSEQWTLPPFLPKRLLLSLLILLAPNAVFWVLALLTATARPIVNLDYLPAALLI 10 20 30 40 50 60 70 80 90 100 110 120 orf48-1. pep ALPWRFVKIAGVLAFWLAVLFDGLMMVIQLFPFMDLIGAINLVPFILTAPAPYQIMTGLL II I II II I II II I II I I II I I I I II I I II I I II II II II II I I I II I I I II I II I I II I orf48ng-l ALPWRFVKIAGVLAFWPAVLFDGLMMVIQLFPFMDLIGAINLVPFILTAPAPYQIMTGLL 70 80 90 100 110 120 130 140 150 160 170 180 orf48-1. pep LLYMLAMPFVLQKAAAKTDFRHIAVCAAWAAAGYFTGHLSYYDRGRMANIFGANNFYYA I II I I I I II II I I I I : I II I II I I I II I II I I I I II II I I II II I II I I II I I I I I I II I orf48ng-l LLYMLAMPFVLQKAAVKTDFRHIAVCAAWAAAGYFTGHLSYYDRGRMANIFGANNFYYA 130 140 150 160 170 180 190 200 210 220 230 240 orf48-1. pep KSQAMLYTVSQNADFITAGLVDPVFLPLGNQQRAATHLNEPKSQKILFIVAESWGLPANP II I I II II I I II I I I I I II I I I I I I II II I I II I I I : I : I II II I I I I I II II I I I I : I I orf48ng-l KSQAMLYTVSQNADFITAGLVDPVFLPLGNQQRAATRLSEPKSQKILFIVAESWGLPGNP 190 200 210 220 230 240 250 260 270 280 290 300 orf48-1. pep ELQNATFAKLLAQKDRFSVWESGSFPFIGATVEGEMRELCAYGGLRGFALRRAPDEKFAR II I I I I I II II I I II II II I I I I I II I I I II I I I I I I I I II II II I I I II I I II II I I I I orf48ng-l ELQNATFAKLLAQKDRFSVWESGSFPFIGATVEGEMRELCAYGGLRGFALRRAPDEKFAR 250 260 270 280 290 300 310 320 330 340 350 360 orf48-1. pep CLPNRLKQEGYATFAMHGAGSSLYDRFSWYPRAGFQEIKTAENLIGKKTCAIFGGVCDSE II I I I I I II II I I II II I II I II I II I I I I II I II I : II II II I I I II I II I I I I I II I I orf48ng-l CLPNRLKQEGYATFAMHGAGSSLYDRFSWYPRAGFQKIKTAENLIGKKTCAIFGGVCDSE 310 320 330 340 350 360 370 380 390 400 410 420 orf48-1. pep LFGEVSAFFKKHDKGLFYWMTLTSHADYPESDIFNHRLKCTEYGLPAETDLCRNFSLHTQ I I I I I I I I I II I I II I II I II II I I I I I I I II I I I I I I I I I I I I I I I I I II I I I I I II I I orf48ng-l LFGEVSAFFKKHDKGLFYWMTLTSHADYPESDIFNHRLKCTEYGLPAETDLCRNFSLHTQ 370 380 390 400 410 420 430 440 450 460 470 orf48-1. pep FFDQLADLIQRPEMKGTEVIIVGDHPPPVGNLNETFRYLKQGHVAWLNFKIKX I I II II II I : I II II II I II I II I I II II I II I I I I I I I I I II I I I I : I II II orf48ng-l FFDQLADLIRRPEMKGTEVIIVGDHPPPVGNLNETFRYLKQGHVAWLHFKIKX 430 440 450 460 470 Based on this analysis, including the presence of a putative leader sequence (double-underlined) and two putative transmembrane domains (single-underlined) in the gonococcal protein, it is predicted that the proteins from N. meningitidis and N.gonorrhoeae, and their epitopes, could be useful antigens for vaccines or diagnostics, or for raising antibodies. Example 57 The following partial DNA sequence was identified in N. meningitidis <SEQ ED 477>: 1 .. GTGAGCGGAC GTTACCGCGC TTTGGATCGC GTTTCCAAAA TCATCATCGT 51 TACTTTGAGT ATCGCCACGC TTGCCGCCGC CGGCATCGCT ATGTCGCGCG 101 GTATGCAGAT GCAGTCCGAT TTTATCGAGC CGACACCGTG GACGCTTGCC 151 GGTTTGGGCT TCCTGATCGC GCTGATGGGC TGGATGCCCG CGCCGATTGA 201 AATTTCCGCC ATCAATTCTT TGTGGGTAAC CGAAAAACAA CGCATCAATC 251 CTTCCGAATA CCGCGACGGG ATTTTTGAAT TCAACGTCGG TTATATCGCC 301 AGTGCGGTTT TGGCTTTGGT TTTCCTTGCA CTGGGCGC.G TAGCGCCGAA 351 CGGCAACGGC GA.ACAGTGC AGATGGCGGG CGGCAAATAT AACGGGCAAT 401 TGATCAATAT GTACGCC .. This coπesponds to the amino acid sequence <SEQ ID 478; ORF53>: 1 ..VSGRYRALDR VSKIIIVTLS IATLAAAGIA MSRGMQMQSD FIEPTPWTLA 51 GLGFLIALMG WMPAPIEISA INSLWVTEKQ RINPSEYRDG IFEFNVGYIA 101 SAVLALVFLA LGXVAPNGNG XTVQMAGGKY NGQLINMYA.. Further work revealed the complete nucleotide sequence <SEQ ID 479>: 1 ATGTCCGAAC AACATATTTC GACTTGGAAA AGTAAAATCA ACGCATTGGG 51 TCCGGGGATC ATGATGGCTT CGGCGGCGGT CGGCGGTTCG CACCTGATTG 101 CCTCGACGCA GGCGGGCGCG CTTTACGGCT GGCAGATCGC GCTCATCATC 151 ATCCTGACCA ACCTCTTCAA ATACCCGTTT TTCCGCTTCA GCGCGCATTA 201 CACGCTGGAC ACGGGCAAGA GCCTGATTGA AGGTTATGCC GAGAAAAGCC 251 GCGTTTATTT GTGGGTATTC CTGATTTTGT GCATCCTCTC CGCCACGATT 301 AACGCGGGCG CGGTCGCCAT TGTAACCGCC GCCATCGTCA AAATGGCGAT 351 TCCCTCGCTG ATGTTTGATG CCGGCACGGT TGCCGCCTTG ATTATGGCAT 401 CCTGCCTGAT TATTTTGGTG AGCGGACGTT ACCGCGCTTT GGATCGCGTT 451 TCCAAAATCA TCATCGTTAC TTTGAGTATC GCCACGCTTG CCGCCGCCGG 501 CATCGCTATG TCGCGCGGTA TGCAGATGCA GTCCGATTTT ATCGAGCCGA 551 CACCGTGGAC GCTTGCCGGT TTGGGCTTCC TGATCGCGCT GATGGGCTGG 601 ATGCCCGCGC CGATTGAAAT TTCCGCCATC AATTCTTTGT GGGTAACCGA 651 AAAACAACGC ATCAATCCTT CCGAATACCG CGACGGGATT TTTGATTTCA 701 ACGTCGGTTA TATCGCCAGT GCGGTTTTGG CTTTGGTTTT CCTTGCACTG 751 GGCGCGTTTG TGCAATACGG CAACGGCGAA GCAGTGCAGA TGGCGGGCGG 801 CAAATATATC GGGCAATTGA TCAATATGTA CGCCGTTACC ATCGGCGGCT 851 GGTCGCGCCC GCTGGTGGCG TTTATCGCGT TTGCCTGTAT GTACGGCACG 901 ACGATTACCG TCGTGGACGG CTATGCCCGT GCCATTGCCG AACCCGTGCG 951 CCTGCTGCGC GGAAAAGACA AAACGGGCAA CGCCGAATTC TTTGCCTGGA 1001 ATATTTGGGT GGCGGGCAGC GGTTTGGCGG TGATTTTCTG GTTTGACGGC 1051 GTAATGGCGA ATCTGCTCAA ATTTGCGATG ATTGCCGCTT TTGTGTCCGC 1101 CCCTGTGTTT GCCTGGCTGA ATTACCGTTT GGTTAAAGGT GATGAAAAAC 1151 ACAAACTCAC ATCAGGTATG AATGCCCTTG CATTGGCAGG CTTGATTTAT 1201 CTGACCGGTT TTACCGTTTT GTTCTTATTG AATTTGGCGG GAATGTTCAA 1251 ATGA This coπesponds to the amino acid sequence <SEQ ID 480; ORF53-l>: 1 MSEQHISTWK SKINALGPGI MMASAAVGGS HLIASTQAGA LYGWQIALII 51 ILTNLFKYPF FRFSAHYTLD TGKSLIEGYA EKSRVYLWVF LILCILSATI 101 NAGAVAIVTA AIVKMAIPSL MFDAGTVAAL IMASCLIILV SGRYRALDRV 151 SKIIIVTLSI ATLAAAGIAM SRGMQMQSDF IEPTPWTLAG LGFLIALMGW 201 MPAPIEISAI NSLWVTEKQR INPSEYRDGI FDFNVGYIAS AVLALVFLAL 251 GAFVQYGNGE AVQMAGGKYI GQLINMYAVT IGGWSRPLVA FIAFACMYGT 301 TITWDGYAR AIAEPVRLLR GKDKTGNAEF FAWNIWVAGS GLAVIFWFDG 351 VMANLLKFAM IAAFVSAPVF AWLNYRLVKG DEKHKLTSGM NALALAGLIY 401 LTGFTVLFLL NLAGMFK* Computer .analysis of this amino acid sequence gave the following results: Homology with a predicted ORF from N.meninsitidis (sfrain Al ORF53 shows 93.5% identity over a 139aa overlap with an ORF (ORF53a) from strain A of N. meningitidis: 10 20 30 orf53. pep VSGRYRALDRVSKIIIVTLSIATLAAAGIA II I I II I I I I II I II II I I I I I II I II II I orf53a AAIVKMAIPSLMFDAGTVAALIMASCLIILVSGRYRALDRVSKIIIVTLSIATLA4AAGIA 110 120 130 140 150 160 40 50 60 70 80 90 orf53. pep MSRGMQMQSDFIEPTPWTLAGLGFLIALMGWMPAPIEISAINSLWVTEKQRINPSEYRDG I II I I II II II II II I I II II I II I I II I II II I I II I I I I I I I I I II I I II II I II II I orf53a MSRGMQMQSDFIEPTPWTLAGLGFLIALMGWMPAPIEISAINSLWVTEKQRINPSEYRDG 170 180 190 200 210 220 100 110 120 130 139 orf53.pep IFEFNVGYIASAVLALVFLALGXVAPNGNGXTVQMAGGKYNGQLINMYA II : I I I I II II II II I I II I I I : II I : II I I I I II I I II II I I orf53a IFDFNVGYIASAVLALVFLALGAFVQYGNGEAVQMAGGKYIGQLINMYAVTIGGWSRPLV 230 240 250 260 270 280 orf53a AFIAFACMYGTTITWDGYARAIAEPVRLLRGKDKTGNAEFFAWNIWVAGSGLAVIFWFD 290 300 310 320 330 340 The complete length ORF53a nucleotide sequence <SEQ ID 481> is: 1 ATGTCCGAAC AACATATTTC GACTTGGAAA AGTAAAATCA ACGCATTGGG 51 ACCGGGGATT ATGATGGCTT CGGCGGCGGT CGGCGGTTCG CACCTGATTG 101 CCTCGACGCA GGCGGGCGCG CTTTACGGCT GGCAGATCGC GCTCATCATC 151 ATCCTGACCA ACCTCTTCAA ATACCCGTTT TTCCGCTTCA GCGCGCATTA 201 CACGCTGGAC ACGGGCAAGA GCCTGATTGA AGGTTATGCC GAGAAAAGCC 251 GCGTTTATTT GTGGGTATTC CTGATTTTGT GCATCCTCTC CGCCACGATT 301 AACGCGGGCG CGGTCGCCAT TGTAACCGCC GCCATCGTCA AAATGGCGAT 351 TCCCTCGCTG ATGTTTGATG CCGGCACGGT TGCCGCCTTG ATTATGGCAT 401 CCTGCCTGAT TATTTTGGTG AGCGGACGTT ACCGCGCTTT GGATCGCGTT 451 TCCAAAATCA TCATCGTTAC TTTGAGTATC GCCACGCTTG CCGCCGCCGG 501 CATCGCTATG TCGCGCGGTA TGCAGATGCA GTCCGATTTT ATCGAGCCGA 551 CACCGTGGAC GCTTGCCGGT TTGGGCTTCC TGATCGCGCT GATGGGCTGG 601 ATGCCCGCGC CGATTGAAAT TTCCGCCATC AATTCTTTGT GGGTAACCGA 651 AAAACAACGC ATCAATCCTT CCGAATACCG CGACGGGATT TTTGATTTCA 701 ACGTCGGTTA TATCGCCAGT GCGGTTTTGG CTTTGGTTTT CCTTGCACTG 751 GGCGCGTTTG TGCAATACGG CAACGGCGAA GCAGTGCAGA TGGCGGGCGG 801 CAAATATATC GGGCAATTGA TCAATATGTA CGCCGTTACC ATCGGCGGCT 851 GGTCGCGCCC GCTGGTGGCG TTTATCGCGT TTGCCTGTAT GTACGGCACG 901 ACGATTACCG TTGTGGACGG CTATGCCCGT GCCATTGCCG AACCCGTGCG 951 CCTGCTGCGC GGAAAAGACA AAACGGGCAA CGCCGAATTC TTTGCCTGGA 1001 ATATTTGGGT GGCGGGCAGC GGTTTGGCGG TGATTTTCTG GTTTGACGGC 1051 GTAATGGCGA ATCTGCTCAA ATTTGCGATG ATTGCCGCTT TTGTGTCCGC 1101 CCCTGTGTTT GCCTGGCTGA ATTACCGTTT GGTCAAAGGT GATGAAAAAC 1151 ACAAACTCAC ATCAGGTATG AATGCCCTTG CATTGGCAGG CTTGATTTAT 1201 CTGACCGGTT TTACCGTTTT GTTCTTATTG AATTTGGCGG GAATGTTCAA 1251 ATGA This encodes a protein having amino acid sequence <SEQ ID 482>: 1 MSEQHISTWK SKINALGPGI MMASAAVGGS HLIASTQAGA LYGWQIALII 51 ILTNLFKYPF FRFSAHYTLD TGKSLIEGYA EKSRVYLWVF LILCILSATI 101 NAGAVAIVTA AIVKMAIPSL MFDAGTVAAL IMASCLIILV SGRYRALDRV 151 SKIIIVTLSI ATLAAAGIAM SRGMQMQSDF IEPTPWTLAG LGFLIALMGW 201 MPAPIEISAI NSLWVTEKQR INPSEYRDGI FDFNVGYIAS AVLALVFLAL 251 GAFVQYGNGE AVQMAGGKYI GQLINMYAVT IGGWSRPLVA FIAFACMYGT 301 TITWDGYAR AIAEPVRLLR GKDKTGNAEF FAWNIWVAGS GLAVIFWFDG 351 VMANLLKFAM IAAFVSAPVF AWLNYRLVKG DEKHKLTSGM NALALAGLIY 401 LTGFTVLFLL NLAGMFK* ORF 53a shows 100.0% identity in 417 aa overlap with ORF53-1: 10 20 30 40 50 60 orf53a. pep MSEQHISTWKSKINALGPGIMMASAAVGGSHLIASTQAGALYGWQIALIIILTNLFKYPF I I I II I I I I I I I I I I I I I I I I I II I I I I I II I I I II I II I I I I I I I I I I I I II I II I I I I orf53-1 MSEQHISTWKSKINALGPGIMMASAAVGGSHLIASTQAGALYGWQIALIIILTNLFKYPF 10 20 30 40 50 60 70 80 90 100 110 120 orf53a . pep FRFSAHYTLDTGKSLIEGYAEKSRVYLWVFLILCILSATINAGAVAIVTAAIVKMAIPSL I I II II I II I II I I I II II II I II II I II I II II II I I I I I II I I I I I I I I II I I I I I I I orf53-1 FRFSAHYTLDTGKSLIEGYAEKSRVYLWVFLILCILSATINAGAVAIVTAAIVKMAIPSL 70 80 90 100 110 120 130 140 150 160 170 180 orf53a . pep MFDAGTVAALIMASCLIILVSGRYRALDRVSKIIIVTLSIATLAAAGIAMSRGMQMQSDF I II I I I I I I I I I I I I II II II I II I II II I I I I I I II I II I I I II II I I II I I II I II II orf53-1 MFDAGTVAALIMASCLIILVSGRYRALDRVSKIIIVTLSIATLAAAGIAMSRGMQMQSDF 130 140 150 160 170 180 190 200 210 220 230 240 orf53a. pep IEPTPWTLAGLGFLIALMGWMPAPIEISAINSLWVTEKQRINPSEYRDGIFDFNVGYIAS II I II I II II I II II II II I I I II I II II II I I II II II I I I II I I I II I I II I I I I I I I orf53-1 IEPTPWTLAGLGFLIALMGWMPAPIEISAINSLWVTEKQRINPSEYRDGIFDFNVGYIAS 190 200 210 220 230 240 250 260 270 280 290 300 orf53a . pep AVLALVFLALGAFVQYGNGEAVQMAGGKYIGQLINMYAVTIGGWSRPLVAFIAFACMYGT I I II I II I I I II II I I I I I I II I I II I II I I I I II I I I I II I I I II I I I I II II I I II I I orf53-1 AVLALVFLALGAFVQYGNGEAVQMAGGKYIGQLINMYAVTIGGWSRPLVAFIAFACMYGT 250 260 270 280 290 300 310 320 330 340 350 360 orf53a. pep TITWDGYARAIAEPVRLLRGKDKTGNAEFFAWNIWVAGSGLAVIFWFDGVMANLLKFAM I I II I I I II I I I II I I I I I I I I I I I II I I II II I I I II I I I II I I I II I I II II II II II orf53-1 TITWDGYARAIAEPVRLLRGKDKTGNAEFFAWNIWVAGSGLAVIFWFDGVMANLLKFAM 310 320 330 340 350 360 370 380 390 400 410 orf53a . pep IAAFVSAPVFAWLNYRLVKGDEKHKLTSGMNALALAGLIYLTGFTVLFLLNLAGMFKX I II I I I I I I I I I I I I I I I I I I I I I I II I I II II I II II I I I II I I I I I I II I I II I II orf53-1 IAAFVSAPVFAWLNYRLVKGDEKHKLTSGMNALALAGLIYLTGFTVLFLLNLAGMFKX 370 380 390 400 410 Homology with a predicted ORF from N.sonorrhoeae ORF53 shows 92.1% identity over a 139aa overlap with a predicted ORF (ORF53ng) from N. gonorrhoeae: orf53. pep VSGRYRALDRVSKIIIVTLSIATLAAAGIA 30 I II II I II I I I II I I I II I II II I II I I II orf53ng AAIVKMAIPSLMFDAGTVAALIMASCLIILVSGRYRALDRVSKIIIVTLSIATLAAAGIA 91 orf53.pep MSRGMQMQSDFIEPTPWTLAGLGFLIALMGWMPAPIEISAINSLWVTEKQRINPSEYRDG 90 II I II I I I II I I I I I I I I I I I II I I II II I I II I II II I I I II I I I I I I II II I I II II orf53ng MSRGMQMQPDFIEPTPWTLAGLGFLIALMGWMPAPIEISAINSLWVTEKQRINPSEYRDG 151 orf53. pep IFEFNVGYIASAVLALVFLALGXVAPNGNGXTVQMAGGKYNGQLINMYA 139 I I : I I I I II I II II I I II I I I I : III : I I I : II II II II II II orf53ng IFDFNVGYIASAVLALVFLALGAFVQYGNGEAVQMGGGKYIGQLINMYAVTIGGGSRPLV 211 An ORF53ng nucleotide sequence <SEQ ED 483> was predicted to encode a protein having amino acid sequence <SEQ ID 484>: 1 MPKKSCVYLW VFLILCIASA TINAGAVAIV TAAIVKMAIP SLMFDAGTVA 51 ALIMASCLII LVSGRYRALD RVSKIIIVTL SIATLAAAGI AMSRGMQMQP 101 DFIEPTPWTL AGLGFLIALM GWMPAPIEIS AINSLWVTEK QRINPSEYRD 151 GIFDFNVGYI ASAVLALVFL ALGAFVQYGN GEAVQMGGGK YIGQLINMYA 201 VTIGGGSRPL VAFIAFACMY GAASTWDGY ARAIAEPVRL LRGKDKTARP 251 IVLLEKLGGR HRFGRDFLV* Further analysis revealed further partial DNA gonococcal sequence <SEQ ID 485>: 1 .. aagaAAAGCT GCGTTTATTT GTGGGTTTTT TTGATTTTGT GTATCGCCTC 51 CGCCACGATT AACGCGGGCG CGGTCGCCAT TGTAACCGCC GCCATCGTCA 101 AAATGGCGAT TCCCTCGCTG ATGTTTGATG CCGGCACGGT TGCCGCCTTG 151 ATTATGGCAT CCTGCCTGAT TATTTTGGTG AGCGGACGTT ACCGCGCTTT 201 GGATCGTGTT TCCAAAATCA TCATTGTTAC TTTGAGCATC GCCACGCTTG 251 CCGCCGCCGG CATCGCTATG TCGCGCGGTA TGCAGATGCA GCCCGATTTT 301 ATCGAGCCGA CACCGTGGAC GCTTGCCGGT TTGGGCTTCC TGATCGCGCT 351 GATGGGCTGG ATGCCCGCGC CGATCGAAAT TTCCGCCATC AATTCTTTGT 401 GGGTAACCGA AAAACAACGC ATCAATCCTT CTGAATACCG CGACGGGATT 451 TTCGATTTCA ACGTCGGTTA TATCGCcagT GCGGTTTTGG CTTTGGTTTT 501 CCTTGCACTG GGCGCGTTTG TGCAATACGG CAACGGCGAA GCAGTGCAGA 551 TGGCGGGCGG CAAATATATC GGGCAATTGA TTAATATGTA TGCCGTAACC 601 ATCGGCGGCT GGTCTCGTCC GCTGGTGGCG TTTATCGCGT TTGCCTGTAT 651 GTACGGCACG ACGATTACCG TTGTGGACGG TTATGCGCGT GCCATTGCCG 701 AACCCGTGCG CCTGCTGCGC GGCAGGGATA AAACCGGCAA CGCCGAGTTG 751 TTtgccTGGA ATATTTGGGT GGCGGGCAGC GGTTTGGCGG TGATTTTCTG 801 GTTTGACggc gcaaTGGCgG AACtgcTCAA ATTTGCGATG ATtgccgcCT 851 TTGTGTCCGC CCCTGTGTTC GCCTGGCTCA ACTACCGCCT CGTCAAAGGG 901 GACAAACGCC ACAGGCTTAC CGCCGGTATG AACGCCCTTG CCATTGTCGG 951 CCTGCTCTAC CTGGCCGGGT TTGCCGTTTT GTTCCTGTTG AACCTTACCG 1001 GACTTTTGGC ATAG This coπesponds to the .amino acid sequence <SEQ ID 486; ORF53ng-l > • 1 ..KKSCVYLWVF LILCIASATI NAGAVAIVTA AIVKMAIPSL MFDAGTVAAL 51 IMASCLIILV SGRYRALDRV SKIIIVTLSI ATLAAAGIAM SRGMQMQPDF 101 IEPTPWTLAG LGFLIALMGW MPAPIEISAI NSLWVTEKQR INPSEYRDGI 151 FDFNVGYIAS AVLALVFLAL GAFVQYGNGE AVQMAGGKYI GQLINMYAVT 201 IGGWSRPLVA FIAFACMYGT TITWDGYAR AIAEPVRLLR GRDKTGNAEL 251 FAWNIWVAGS GLAVIFWFDG AMAELLKFAM IAAFVSAPVF AWLNYRLVKG 301 DKRHRLTAGM NALAIVGLLY LAGFAVLFLL NLTGLLA* ORF53ng-l and ORF53-1 show 94.0% identity in 336 aa overlap: 60 70 80 90 100 110 orf53-1. pep ILTNLFKYPFFRFSAHYTLDTGKSLIEGYAEKSRVYLWVFLILCILSATINAGAVAIVTA : I I II II II I I I I I II I I II II I II I I I orf53ng-l KKSCVYLWVFLILCIASATINAGAVAIVTA 10 20 30 120 130 140 150 160 170 orf53-1. pep AIVKMAIPSLMFDAGTVAALIMASCLIILVSGRYRALDRVSKIIIVTLSIATLAAAGIAM II I I II I II I II II I II II I I II I I II I I I II I I II I II I I I I II I II I I II II II II II orf53ng-l AIVKMAIPSLMFDAGTVAALIMASCLIILVSGRYRALDRVSKIIIVTLSIATLAAAGIAM 40 50 60 70 80 90 180 190 200 210 220 230 orf53-1. pep SRGMQMQSDFIEPTPWTLAGLGFLIALMGWMPAPIEISAINSLWVTEKQRINPSEYRDGI I II II I I I II I I I II I II I II I I I II I I II I I II I I I I I I I I II I II I I II II I I I II I orf53ng-l SRGMQMQPDFIEPTPWTLAGLGFLIALMGWMPAPIEISAINSLWVTEKQRINPSEYRDGI 100 110 120 130 140 150 240 250 260 270 280 290 orf53-1. pep FDFNVGYIASAVLALVFLALGAFVQYGNGEAVQMAGGKYIGQLINMYAVTIGGWSRPLVA II I I II I II I II II I II I I I I I I II I II II I II I I II I I I II I I I I II I I I I II I I II I I orf53ng-l FDFNVGYIASAVLALVFLALGAFVQYGNGEAVQMAGGKYIGQLINMYAVTIGGWSRPLVA 160 170 180 190 200 210 300 310 320 330 340 350 orf53-1. pep FIAFACMYGTTITWDGYARAIAEPVRLLRGKDKTGNAEFFAWNIWVAGSGLAVIFWFDG I I II I I I I II I I I I I I I II I I II I I I I I I I I : I I II I II : I I I I II I I I I I II I I II I I I orf53ng-l FIAFACMYGTTITWDGYARAIAEPVRLLRGRDKTGNAELFAWNIWVAGSGLAVIFWFDG 220 230 240 250 260 270 360 370 380 390 400 410 orf53-1. pep VMANLLKFAMIAAFVSAPVFAWLNYRLVKGDEKHKLTSGMNALALAGLIYLTGFTVLFLL M I : I II I I II I I I I I I I II I I I II I II I I I :: I : I I : I I II I I :: II : I I : II : I I II I orf53ng-l AMAELLKFAMIAAFVSAPVFAWLNYRLVKGDKRHRLTAGMNALAIVGLLYLAGFAVLFLL 280 290 300 310 320 330 orf53-1. pep NLAGMFKX I I: I :: orf53ng-l NLTGLLAX Based on this analysis, including the presence of a putative leader sequence (double-underlined) and several putative transmembrane domains (single-underlined) in the gonococcal protein, it is predicted that the proteins from N. meningitidis and N.gonorrhoeae, and their epitopes, could be useful antigens for vaccines or diagnostics, or for raising antibodies. Example 58 The following partial DNA sequence was identified in N. meningitidis <SEQ ID 487>: 1 .. TGCGGGAAA CGGCATATGT TTTGGATAGT TTTGATCGTT ATTTTGTTGT 51 TGCGCTTGCC GGCTTGTTTT TTGTCCGCGC ACAATCCGAA CGCGAGTGGA 101 TGCGCGAGGT TTCTGCGTGG CAGGAAAAGA AAGGGGAAAA ACAGGCGGAG 151 CTGCCTGAAA TCAAAGACGG TATGCCCGAT TTTCCCGAAC TTGCCCTGAT 201 GCTTTTCCAC GCCGTCAAAA CGGCAGTGTA TTGGCTGTTT GTCGGTGTCG 251 TCCGTTTCTG CCGAAACTAT CTGGCGCACG AATCCGAACC GGACAGGCCC 301 GTTCCGCCT.. This coπesponds to the amino acid sequence <SEQ ID 488; ORF58>: 1 .. LRETAYVLDS FDRYFWAiA GLFFVRAQSE REWMREVSAW QEKKGEKQAE 51 LPEIKDGMPD FPELALMLFH AVKTAVYWLF VGWRFCRNY LAHESEPDRP 101 VPP.. Further work revealed the complete nucleotide sequence <SEQ ID 489>: 1 ATGTTTTGGA TAGTTTTGAT CGTTATTTTG TTGCTTGCGC TTGCCGGCTT 51 GTTTTTTGTC CGCGCACAAT CCGAACGCGA GTGGATGCGC GAGGTTTCTG 101 CGTGGCAGGA AAAGAAAGGG GAAAAACAGG CGGAGCTGCC TGAAATCAAA 151 GACGGTATGC CCGATTTTCC CGAACTTGCC CTGATGCTTT TCCATGCCGT 201 CAAAACGGCA GTGTATTGGC TGTTTGTCGG TGTCGTCCGT TTCTGCCGAA 251 ACTATCTGGC GCACGAATCC GAACCGGACA GGCCCGTTCC GCCTGCTTCT 301 GCAAACCGTG CGGATGTTCC GACCGCATCC GACGGATATT CAGACAGTGG 351 AAACGGGACG GAAGAAGCGG AAACGGAAGA AGCAGAAGCT GCGGAGGAAG 401 AGGCTGCCGA TACGGAAGAC ATTGCAACTG CCGTAATCGA CAACCGCCGC 451 ATCCCATTCG ACCGGAGTAT TGCTGAAGGG TTGATGCCGT CTGAAAGCGA 501 AATTTCGCCC GTCCGTCCGG TTTTTAAAGA AATCACTTTG GAAGAAGCAA 551 CGCGTGCTTT AAACAGCGCG GCTTTAAGGG AAACGAAAAA ACGCTATATC 601 GATGCATTTG AGAAAAACGA AACAGCGGTC CCCAAAGTCC GCGTGTCCGA 651 TACCCCGATG GAAGGGCTGC AGATTATCGG TTTGGACGAC CCTGTGCTTC 701 AACGCACGTA TTCCCATATG TTCGATGCGG ACAAAGAAGC GTTTTCCGAG 751 TCTGCGGATT ACGGATTTGA GCCGTATTTT GAGAAGCAGC ATCCGTCTGC 801 CTTTTCTGCA GTCAAAGCCG AAAATGCACG GAATGCGCCG TTCCACCGTC 851 ATGCAGGGCA GGGGAAAGGG CAGGCGGAGG CAAAATCCCC GGATGTTTCC 901 CAAGGGCAGT CCGTTTCAGA CGGCACGGCC GTCCGCGATG CCCGCCGCCG 951 CGTTTCCGTC AATTTGAAAG AACCGAACAA GGCAACGGTT TCTGCGGAGG 1001 CGCGAATTTC TCGCCTGATT CCGGAAAGTC AGACGGTTGT CGGGAAACGG 1051 GATGTCGAAA TGCCGTCTGA AACCGAAAAT GTTTTCACGG AAACCGTTTC 1101 GTCTGTGGGA TACGGCGGTC CGGTTTATGA TGAAACTGCC GATATCCATA 1151 TTGAAGAACC TGCCGCGCCC GATGCTTGGG TGGTCGAACC ACCCGAAGTG 1201 CCGAAAGTTC CCATGACCGC AATCGATATT CAGCCGCCGC CTCCCGTATC 1251 GGAAATCTAC AACCGTACCT ATGAACCGCC GTCAGGATTC GAGCAGGTGC 1301 AACGCAGCCG CATTGCCGAG ACCGACCATC TTGCCGATGA TGTTTTGAAT 1351 GGAGGTTGGC AGGAGGAAAC CGCCGCTATT GCGGATGACG GCAGTGAAGG 1401 TGCGGCAGAG CGGTCAAGCG GGCAATATCT GTCGGAAACC GAAGCGTTCG 1451 GGCATGACAG TCAGGCGGTT TGTCCGTTTG AAAATGTGCC GTCTGAACGC 1501 CCGTCCTGCC GGGTATCGGA TACGGAAGCG GATGAAGGGG CGTTCCCATC 1551 TGAAGAAACC GGTGCGGTAT CCGAACACCT GCCGACAACC GACCTGCTTC 1601 TGCCTCCGCT GTTCAATCCC GAGGCGACGC AAACCGAAGA AGAACTGTTG 1651 GAAAACAGCA TCACCATCGA AGAAAAATTG GCGGAGTTCA AAGTCAAGGT 1701 CAAGGTTGTC GATTCTTATT CCGGCCCCGT AATTACGCGT TATGAAATCG 1751 AACCCGATGT CGGCGTGCGC GGCAATTCCG TTCTGAATCT GGAAAAAGAT 1801 TTGGCGCGTT CGCTCGGCGT GGCTTCCATC CGCGTTGTCG AAACCATCCC 1851 CGGCAAAACC TGCATGGGTT TGGAACTTCC GAACCCGAAA CGCCAAATGA 1901 TACGCCTGAG CGAAATCTTC AATTCGCCCG AGTTTGCCGA ATCCAAATCC 1951 AAGCTGACGC TCGCGCTCGG TCAGGACATC ACCGGACAGC CCGTCGTAAC 2001 CGACTTGGGA AAAGCACCGC ATTTGTTGGT TGCCGGCACG ACCGGTTCGG 2051 GCAAATCGGT GGGTGTCAAC GCGATGATTC TGTCTATGCT TTTCAAAGCC 2101 GCGCCGGAAG ACGTGCGTAT GATTATGATC GATCCGAAAA TGCTGGAATT 2151 GAGCATTTAC GAAGGCATCC CGCACCTGCT CGCCCCTGTC GTTACCGATA 2201 TGAAGCTGGC GGCAAACGCG CTGAACTGGT GTGTTAACGA AATGGAAAAA 2251 CGCTACCGCC TGATGAGCTT TATGGGCGTG CGTAATCTTG CGGGCTTCAA 2301 TCAAAAAATC GCCGAAGCCG CAGCAAGGGG AGAAAAAATC GGCAATCCGT 2351 TCAGCCTCAC GCCCGACGAT CCCGAACCTT TGGAAAAACT GCCGTTTATC 2401 GTGGTCGTGG TCGATGAGTT TGCCGACCTG ATGATGACGG CAGGCAAGAA 2451 AATCGAAGAA CTGATTGCCC GCCTCGCCCA AAAAGCCCGC GCGGCAGGCA 2501 TCCATTTGAT TCTTGCCACA CAACGCCCCA GCGTCGATGT CATCACGGGT 2551 CTGATTAAGG CGAACATCCC GACGCGTATC GCGTTCCAAG TGTCCAGCAA 2601 AATCGACAGC CGCACGATTC TCGACCAAAT GGGCGCGGAA AACCTGCTCG 2651 GTCAGGGCGA TATGCTGTTC CTGCTGCCGG GTACTGCCTA TCCGCAGCGC 2701 GTTCACGGCG CGTTTGCCTC GGATGAAGAG GTGCACCGCG TGGTCGAATA 2751 TTTGAAACAG TTTGGCGAAC CGGACTATGT TGACGATATT TTGAGCGGCG 2801 GCGGCAGCGA AGAGCTGCCC GGCATCGGGC GCAGCGGCGA CGACGAAACC 2851 GATCCGATGT ACGACGAGGC CGTATCCGTT GTCCTGAAAA CGCGCAAAGC 2901 CAGCATTTCG GGCGTACAGC GCGCCTTGCG TATCGGCTAC AACCGCGCCG 2951 CGCGTCTGAT TGACCAGATG GAGGCGGAAG GCATTGTGTC CGCACCGGAA 3001 CACAACGGCA ACCGTACGAT TCTCGTCCCC TTGGACAATG CTTGA This coπesponds to the amino acid sequence <SEQ ED 490; ORF58-1 > • 1 MFWIVLIVIL LLAAGLFFV RAQSEREWMR EVSAWQEKKG EKQAELPEIK 51 DGMPDFPELA LMLFHAVKTA VYWLFVGWR FCRNYLAHES EPDRPVPPAS 101 ANRADVPTAS DGYSDSGNGT EEAETEEAEA AEEEAADTED IATAVIDNRR 151 IPFDRSIAEG LMPSESEISP VRPVFKEITL EEATRALNSA ALRETKKRYI 201 DAFEKNETAV PKVRVSDTPM EGLQIIGLDD PVLQRTYSHM FDADKEAFSE 251 SADYGFEPYF EKQHPSAFSA VKAENARNAP FHRHAGQGKG QAEAKSPDVS 301 QGQSVSDGTA VRDARRRVSV NLKEPNKATV SAEARISRLI PESQTWGKR 351 DVEMPSETEN VFTETVSSVG YGGPVYDETA DIHIEEPAAP DAWWEPPEV 401 PKVPMTAIDI QPPPPVSEIY NRTYEPPSGF EQVQRSRIAE TDHLADDVLN 451 GGWQEETAAI ADDGSEGAAE RSSGQYLSET EAFGHDSQAV CPFENVPSER 501 PSCRVSDTEA DEGAFPSEET GAVSEHLPTT DLLLPPLFNP EATQTEEELL 551 ENSITIEEKL AEFKVKVKW DSYSGPVITR YEIEPDVGVR GNSVLNLEKD 601 LARSLGVASI RWETIPGKT CMGLELPNPK RQMIRLSEIF NSPEFAESKS 651 KLTLALGQDI TGQPWTDLG KAPHLLVAGT TGSGKSVGVN AMILSMLFKA 701 APEDVRMIMI DPKMLELSIY EGIPHLLAPV VTDMKLAANA LNWCVNEMEK 751 RYRLMSFMGV RNLAGFNQKI AEAAARGEKI GNPFSLTPDD PEPLEKLPFI 801 WWDEFADL MMTAGKKIEE LIARLAQKAR AAGIHLILAT QRPSVDVITG 851 LIKANIPTRI AFQVSSKIDS RTILDQMGAE NLLGQGDMLF LLPGTAYPQR 901 VHGAFASDEE VHRWEYLKQ FGEPDYVDDI LSGGGSEELP GIGRSGDDET 951 DPMYDEAVSV VLKTRKASIS GVQRALRIGY NRAARLIDQM EAEGIVSAPE 1001 HNGNRTILVP LDNA* Computer analysis of this amino acid sequence predicts the indicated transmembrane region, and also gave the following results: Homology with a predicted ORF from N.meninsitidis (sfrain Al ORF58 shows 96.6% identity over a 89aa overlap with an ORF (ORF58a) from strain A of N. meningitidis: 10 20 30 40 50 60 orf 58 . pep LRETAYVLDSFDRYFWALAGLFFVRAQSEREWMREVSAWQEKKGEKQAELPEIKDGMPD : : : I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I orf 58a MFWIVLIVILLLALAGLFFVRAQSEREWMREVSAWQEKKGEKQAELPEIKDGMPD 10 20 30 40 50 70 80 90 100 orf58.pep FPELALMLFHAVKTAVYWLFVGWRFCRNYLAHESEPDRPVPP I II I I I II I II I I I I I I I I I I II I I II I II II II I I II I I I I I orf58a FPELALMLFHAVKTAVYWLFVGWRFCRNYLAHESEPDRPVPPASANRADVPTASDGYSD 60 70 80 90 100 110 The complete length ORF58a nucleotide sequence <SEQ ID 491> is: 1 ATGTTTTGGA TAGTTTTGAT CGTTATTTTG TTGCTTGCGC TTGCCGGCTT 51 GTTTTTTGTC CGCGCACAAT CCGAACGCGA GTGGATGCGC GAGGTTTCTG 101 CGTGGCAGGA AAAGAAAGGG GAAAAACAGG CGGAGCTGCC TGAAATCAAA 151 GACGGTATGC CCGATTTTCC CGAACTTGCC CTGATGCTTT TCCATGCCGT 201 CAAAACGGCA GTGTATTGGC TGTTTGTCGG TGTCGTCCGT TTCTGCCGAA 251 ACTATCTGGC GCACGAATCC GAACCGGACA GGCCCGTTCC GCCTGCTTCT 301 GCAAATCGTG CGGATGTTCC GACCGCATCC GACGGATATT CAGACAGTGG 351 AAACGGGACG GAAGAAGCGG AAACGGAAGA AGCAGAAGCT GCGGAGGAAG 401 AGGCTGCCGA TACGGAAGAC ATTGCAACTG CCGTAATCGA CAACCGCCGC 451 ATCCCATTCG ACCGGAGTAT TGCTGAAGGG TTGATGCCGT CTGAAAGCGA 501 AATTTCGCCC GTCCGTCCGG TTTTTAAGGA AATCACTTTG GAAGAAGCAA 551 CGCGTGCTTT AAACAGCGCG GCTTTAAGGG AAACGAAAAA ACGCTATATC 601 GATGCATTTG AGAAAAACGA AACAGCGGTC CCCAAAGTCC GCGTGTCCGA 651 TACCCCGATG GAAGGGCTGC AGATTATCGG TTTGGACGAC CCTGTGCTTC 701 AACGCACGTA TTCCCGTATG TTCGATGCGG ACAAAGAAGC GTTTTCCGAG 751 TCTGCGGATT ACGGATTTGA GCCGTATTTT GAGAAGCAGC ATCCGTCTGC 801 CTTTTCTGCA GTCAAAGCCG AAAATGCACG GAATGCGCCG TTCCGCCGTC 851 ATGCAGGGCA GGGNAAAGGG CAGGCGGAGG CNAAATCCCC GGATGTTTCC 901 CAAGGGCAGT CCGTTTCAGA CGGCACAGCC GTCCGCGATG CCNGCCGCCG 951 CGTTTCCGTC AATTTGAAAG AACCGAACAA GGCAACGGTT TCTGCGGAGG 1001 CGCGGATTTC GCGCCTGATT CCGGAAAGTC GGACGGTTGT CGGGAAACGG 1051 GATGTCGAAA TGCCGTCTGA AACCGAAAAT GTTTTCACGG AAANTGTTTC 1101 GTCTGTGGGA TACGGCGNTC CGGTTTATGA TGAAACTGCC GATATCCATA 1151 TTGAAGAACC TGCCGCGCCC GATGCTTGGG TGGTCGAACC ACCCGAAGTG 1201 CCGAAAGTTC CCATGCCCGC AATNGATATT CCGCCGCCGC CTCCCGTATC 1251 GGAAATCTAC AACCGTACCT ATGAACCGCC GGCAGGATTC GAGCAGGTGC 1301 AACGCAGCCG CATTGCCGAA ACCGATCATC TTGCCGATGA TGTTTTGAAT 1351 GGAGGTTGGC AGGAGGAAAC CGCCGCTATT GCGAATGACG GCAGTGAGGG 1401 TGTGGCAGAG CGGTCAAGCG GGCAATATTT GTCGGAAACC GAAGCGTTCG 1451 GGCATGACAG TCAGGCGGTT TGTCCGTTTG AAAATGTGCC GTCTGAACGC 1501 CCGTCCCGCC GGGCATNGGA TACGGAAGCG GATGAAGGGG CGTTCCAATC 1551 TGAAGAAACC GGTGCGGTAT CCGAACACCT GCCGACAACC GACCTGCTTC 1601 TGCCGCCGCT GTTCAATCCC GGGGCGACGC AAACCGAAGA AGANCTGTTG 1651 GANAACAGCA TCACCATCGA AGAAAAATNG GCGGAGTTCA AAGTCAAGGT 1701 CAAGGTTGTC GATTCTTATT CCGGCCCCGT GATTACGCGT TATGAAATCG 1751 AACCCGATGT CGGCGTGCGC GGCAATTCCG TTCTAAATCT GGAAAAAGAN 1801 TTGGCGCGTT CGCTCGGCGT GGCTTCCATC CGCGTTGTCG AAACCATCCT 1851 CGGCAAAACC TGTATGGGTT TGGAACTTCC GAACCCGAAA CGCCAAATGA 1901 TACGCCTGAG CGAAATCTTC AATTCGCCCG AGTTTGCCGA ATCCAAATCC 1951 AAGCTGACGC TCGCGCTCGG TCAGGACATC ACCGGACAGC CCGTCGTAAC 2001 CGACTTGGGC AAAGCACCGC ATTTGTTGGT TGCCGGCACG ACCGGTTCGG 2051 GCAAATCGGT GGGTGTCAAC GCGATGATTC TGTCTATGCT TTTCAAAGCC 2101 GCGCCGGAAG ACGTGCGTAT GATTATGATC GATCCGAAAA TGCTGGAATT 2151 GAGCATTTAC GAAGGCATCC CGCACCTGCT CGCCCCTGTC GTTACCGATA 2201 TGAAGCTGGC GGCAAACGCG CTGAACTGGT GTGTTAACGA AATGGAAAAA 2251 CGCTACCGCC TGATGAGCTT TATGGGCGTG CGCAATCTTG CGGGTNTCAA 2301 TCAAAAAATC GCCGAAGCCG CAGCAAGGGG GGAGAAAATC GGCAACCCGT 2351 TCAGCCTCAC GCCCGACAAT CCCGAACCTT TGGANAAATT GCCGTTTATC 2401 GTGGTCGTGG TTGATGAGTT TGCCGACCTG ATGATGACGG CAGGCAAGAA 2451 AATCGAAGAA CTGATTGCCC GCCTCGCCCA AAAAGCCCGC GCGGCAGGCA 2501 TCCATCTTAT CCTTGCCACA CAACGCCCCA GTGTCGATGT CATCACGGGT 2551 CTGATTAAGG CGAACATCCC GACGCGTATC GCGTTCCAAG TGTCCAGCAA 2601 AATCGACAGC CGCACGATTC TTGACCAAAT GGGTGCGGAA AACCTGCTCG 2651 GGCAGGGCGA TATGCTGTTC CTGCCGCCGG GTACGGCCTA TCCGCAGCGC 2701 GTTCACGGCG CGTTTGCCTC GGATGAAGAG GTGCACCGCG TGGTCGAATA 2751 TCTGAAACAG TTTGGCGAAC CGGACTATGT TGACGATATN TTGAGCGGCG 2801 GTATGTCCGA CGATTTGCTG GGAATCAGCC GGAGCGGCGA CGGCGAAACC 2851 GATCCGATGT ACGACGAGGC CGTGTCNGTT GTTTTGAAAA CGCGCAAAGC 2901 CAGCATTTCT GGCGTGCAGC GCGCATTGCG TATCGGCTAT AATCGCGCCG 2951 CGCGTCTGAT TGACCAGATG GAGGCGGAAG GCATTGTGTC CGCACCGGAA 3001 CACAACGGCA ACCGTACGAT TCTCGTCCCC TTNGACAATG CTTGA This encodes a protein having amino acid sequence <SEQ ID 492>: 1 MFWIVLIVIL LLALAGLFFV RAQSEREWMR EVSAWQEKKG EKQAELPEIK 51 DGMPDFPELA LMLFHAVKTA VYWLFVGWR FCRNYLAHES EPDRPVPPAS 101 ANRADVPTAS DGYSDSGNGT EEAETEEAEA AEEEAADTED IATAVIDNRR 151 IPFDRSIAEG LMPSESEISP VRPVFKEITL EEATRALNSA ALRETKKRYI 201 DAFEKNETAV PKVRVSDTPM EGLQIIGLDD PVLQRTYSRM FDADKEAFSE 251 SADYGFEPYF EKQHPSAFSA VKAENARNAP FRRHAGQGKG QAEAKSPDVS 301 QGQSVSDGTA VRDAXRRVSV NLKEPNKATV SAEARISRLI PESRTWGKR 351 DVEMPSETEN VFTEXVSSVG YGXPVYDETA DIHIEEPAAP DAWWEPPEV 401 PKVPMPAXDI PPPPPVSEIY NRTYEPPAGF EQVQRSRIAE TDHLADDVLN 451 GGWQEETAAI ANDGSEGVAE RSSGQYLSET EAFGHDSQAV CPFENVPSER 501 PSRRAXDTEA DEGAFQSEET GAVSEHLPTT DLLLPPLFNP GATQTEEXLL 551 XNSITIEEKX AEFKVKVKW DSYSGPVITR YEIEPDVGVR GNSVLNLEKX 601 LARSLGVASI RWETILGKT CMGLELPNPK RQMIRLSEIF NSPEFAESKS 651 KLTLALGQDI TGQPWTDLG KAPHLLVAGT TGSGKSVGVN AMILSMLFKA 701 APEDVRMIMI DPKMLELSIY EGIPHLLAPV VTDMKLAANA LNWCVNEMEK 751 RYRLMSFMGV RNLAGXNQKI AEAAARGEKI GNPFSLTPDN PEPLXKLPFI 801 WWDEFADL MMTAGKKIEE LIARLAQKAR AAGIHLILAT QRPSVDVITG 851 LIKANIPTRI AFQVSSKIDS RTILDQMGAE NLLGQGDMLF LPPGTAYPQR 901 VHGAFASDEE VHRVVEYLKQ FGEPDYVDDX LSGGMSDDLL GISRSGDGET 951 DPMYDEAVSV VLKTRKASIS GVQRALRIGY NRAARLIDQM EAEGIVSAPE 1001 HNGNRTILVP XDNA* ORF58a and ORF58-1 show 96.6% identity in 1014 aa overlap: 10 20 30 40 50 60 orf58a . pep MFWIVLIVILLLALAGLFFVRAQSEREWMREVSAWQEKKGEKQAELPEIKDGMPDFPELA I I II I I I I I I I I I I I II II I II I II I I I I I II II I II II I I I II II I I I I II I II II II I orf58-1 MFWIVLIVILLLALAGLFFVRAQSEREWMREVSAWQEKKGEKQAELPEIKDGMPDFPELA 10 20 30 40 50 60 70 80 90 100 110 120 orf58a . pep LMLFHAVKTAVYWLFVGWRFCRNYLAHESEPDRPVPPASANRADVPTASDGYSDSGNGT I I II I I I II I II I I II I I I I II I I II I II I I II I I I II II I I I II I I I II I II I I I I I I I orf58-1 LMLFHAVKTAVYWLFVGWRFCRNYLAHESEPDRPVPPAΞANRADVPTASDGYSDSGNGT 70 80 90 100 110 120 130 140 150 160 170 180 orf58a . pep EEAETEEAEAAEEEAADTEDIATAVIDNRRIPFDRSIAEGLMPSESEISPVRPVFKEITL II I II II I II I I I I I II II I I I II II I I I I II I I I I I I I I I I I II II I I II I I I I I I I I I orf58-1 EEAETEEAEAAEEEAADTEDIATAVIDNRRIPFDRSIAEGLMPSESEISPVRPVFKEITL 130 140 150 160 170 180 190 200 210 220 230 240 orf58a. pep EEATRALNSAALRETKKRYIDAFEKNETAVPKVRVSDTPMEGLQIIGLDDPVLQRTYSRM II I II II I II I I I I I I II II II I I II I II I I II I I I I I II II I II I I I I I I II I I I II : I orf58-1 EEATRALNSAALRETKKRYIDAFEKNETAVPKVRVSDTPMEGLQIIGLDDPVLQRTYSHM 190 200 210 220 230 240 250 260 270 280 290 300 orf58a . pep FDADKEAFSESADYGFEPYFEKQHPSAFSAVKAENARNAPFRRHAGQGKGQAEAKSPDVS II I II II I II I I I I I II I II II I I II I II I I II I II I I II I : I II I I I I I I II I I I I I II orf58-1 FDADKEAFSESADYGFEPYFEKQHPSAFSAVKAENARNAPFHRHAGQGKGQAEAKSPDVS 250 260 270 280 290 300 310 320 330 340 350 360 orf58a . pep QGQSVSDGTAVRDAXRRVSVNLKEPNKATVSAEARISRLIPESRTWGKRDVEMPSETEN I I I I I I I II I II II I I II I I I II I I I I I I II I I I I I I I I I I I: I II I II I I I I I I I I I I orf58-1 QGQSVSDGTAVRDARRRVSVNLKEPNKATVSAEARISRLIPESQTWGKRDVEMPSETEN 310 320 330 340 350 360 370 380 390 400 410 420 orf58a. pep VFTEXVSSVGYGXPVYDETADIHIEEPAAPDAWWEPPEVPKVPMPAXDIPPPPPVSEIY II I I : I I I I I I I I I I I I I II I II I I I I I I II I I I I II II I I I II I II II I I I II I I orf58-1 VFTETVSSVGYGGPVYDETADIHIEEPAAPDAWWEPPEVPKVPMTAIDIQPPPPVSEIY 370 380 390 400 410 420 430 440 450 460 470 480 orf58a. pep NRTYEPPAGFEQVQRSRIAETDHLADDVLNGGWQEETAAIANDGSEGVAERSSGQYLSET II I I II I : I I II II I II II I I II I II I II II I II I I I II I I: I I I I I: I II II I I I I I I I orf58-1 NRTYEPPSGFEQVQRSRIAETDHLADDVLNGGWQEETAAIADDGSEGAAERSSGQYLSET 430 440 450 460 470 480 490 500 510 520 530 540 orf58a . pep EAFGHDSQAVCPFENVPSERPSRRAXDTEADEGAFQSEETGAVSEHLPTTDLLLPPLFNP I I I I I I I II I II I I I I I I I I I I I: I I I I II I I I II II II I II I I I I I II II II II II orf58-1 EAFGHDSQAVCPFENVPSERPSCRVSDTEADEGAFPSEETGAVSEHLPTTDLLLPPLFNP 490 500 510 520 530 540 550 560 570 580 590 600 orf58a . pep GATQTEEXLLXNSITIEEKXAEFKVKVKWDSYSGPVITRYEIEPDVGVRGNSVLNLEKX I I II I I II I I I I II II I I I II I I I I II II I II I II I I I I II I II I I I II I II II I orf58-1 EATQTEEELLENSITIEEKLAEFKVKVKWDSYSGPVITRYEIEPDVGVRGNSVLNLEKD 550 560 570 580 590 600 610 620 630 640 650 660 orf58a . pep LARSLGVASIRWETILGKTCMGLELPNPKRQMIRLSEIFNSPEFAESKSKLTLALGQDI I I I II I I I I II I I I I I II I II I II I I I II I I II I II I I II I II II I I I I II II I I I I II orf58-1 LARSLGVASIRWETIPGKTCMGLELPNPKRQMIRLSEIFNSPEFAESKSKLTLALGQDI 610 620 630 640 650 660 670 680 690 700 710 720 orf58a . pep TGQPWTDLGKAPHLLVAGTTGSGKSVGVNAMILSMLFKAAPEDVRMIMIDPKMLELSIY I II II II I II I II II II II I II I II I II II I I I II I I I I I I II I I I I I II I II I II II I I orf58-1 TGQPWTDLGKAPHLLVAGTTGSGKSVGVNAMILSMLFKAAPEDVRMIMIDPKMLELSIY 670 680 690 700 710 720 730 740 750 760 770 780 orf58a . pep EGIPHLLAPVVTDMKLAANALNWCVNEMEKRYRLMSFMGVRNLAGXNQKIAEAAARGEKI I II I I I I I II II I II II II I II I I II II I I I I I I II I II I I I I II I I I I II II I I I I II orf58-1 EGIPHLLAPWTDMKLAANALNWCVNEMEKRYRLMSFMGVRNLAGFNQKIAEAAARGEKI 730 740 750 760 770 780 790 800 810 820 830 840 orf58a . pep GNPFSLTPDNPEPLXKLPFIWWDEFADLMMTAGKKIEELIARLAQKARAAGIHLILAT II I I II I II : I I II I I I I II I I II I II II II I I I I II I I II I II I I I I I II II I I I I II orf58-1 GNPFSLTPDDPEPLEKLPFIWWDEFADLMMTAGKKIEELIARLAQKARAAGIHLILAT 790 800 810 820 830 840 850 860 870 880 890 900 orf58a. pep QRPSVDVITGLIKANIPTRIAFQVSSKIDSRTILDQMGAENLLGQGDMLFLPPGTAYPQR II I I II I II I II II I I I II II I I I II I II I I II I II I II II I II I I I I I I I I II I I I I I orf58-1 QRPSVDVITGLIKANIPTRIAFQVSSKIDSRTILDQMGAENLLGQGDMLFLLPGTAYPQR 850 860 870 880 890 900 910 920 930 940 950 960 orf58a . pep VHGAFASDEEVHRWEYLKQFGEPDYVDDXLSGGMSDDLLGISRSGDGETDPMYDEAVSV I I I II I I II I II II I I I II I II I II I I I I I II I \ : : \ I I: I I I I I I I II II II I I I orf58-1 VHGAFASDEEVHRVVEYLKQFGEPDYVDDILSGGGSEELPGIGRSGDDETDPMYDEAVSV 910 920 930 940 950 960 970 980 990 1000 1010 orf58a . pep VLKTRKASISGVQRALRIGYNRAARLIDQMEAEGIVSAPEHNGNRTILVPXDNAX II I I I I I II I II I II II I II I I II I I I I II II I I II II I I I I I II II I I I I I II orf58-1 VLKTRKASISGVQRALRIGYNRAARLIDQMEAEGIVSAPEHNGNRTILVPLDNAX 970 980 990 1000 1010 Homology with a predicted ORF from Nsonorrhoeae ORF58 shows complete identity over a 9aa overlap with a predicted ORF (ORF58ng) from N. gonorrhoeae: orf 58 . pep ALMLFHAVKTAVYWLFVGWRFCRΝYLAHESEPDRPVPP 103 I I I I I I I I I orf58ng SEPDRPVPPASAΝRADVPTASDGYSDSGΝG 30 The ORF58ng nucleotide sequence <SEQ ED 493> is predicted to encode a protein having partial amino acid sequence <SEQ ID 494>: 1 .. SEPDflPVPPA SAΝRADVPTA SDGYSDSGΝG TEEAETEAAE AAEEEAADTE 51 DIATAVIDΝR RIPFDRSIAE GLMQSESKTS PVRPVFKEIT LEEATRALSS 101 AALRETKKRY IDAFEKΝGTA VPKVRVSDTP MEGLQIIGLD DPVLQRTYSR 151 MFDADKEAFS ESADYGFEPY FEKQHPSAFS AVKAEΝARΝA PFRRHAGQEK 201 GQAEAKSPDV SQGQSVSDGT AVRDARRRVS VΝLKEPΝKAT VSAEARISRL 251 IPESRTWGK RDVEMPSETE ΝVFTETVSSV GYGGPVYDEA ADIHIEEPAA 301 PDAWWEPPE VPEVAVPEID ILPPPPVSEI YΝRTYEPPAG FEQAQRSRIA -291- 351 ETDHLAADVL NGGWQEETAA IADDGSEGAA ERSSGQYLSE TEAFGHDSQA 401 VCPFEDVPSE RPSCRVSDTE ADEGAFQSEE TGAVSEHLPT TDLLLPPLFN 451 PEATQTEEEL LENSITIEEK LAEFKVKVKV VDSYSGPVIT RYEIEPDVGV 501 RGNSVLNLEK DLARSLGVAS IRWETIPGK TCMGLELPNP KRQMIRLSEI 551 FNSPEFAESK SKLTLALGQD ITGQPWTDL GKAPHLLVAG TTGSGKSVGV 601 NAMILSMLFK AAPEDVRMIM IDPKMLELSI YEGITHLLAP WTDMKLAAN 651 ALNWCVNEME KRYRLMSFMG VRNLAGFNQK IAEAAARGEK IGNPFSLTPD 701 DPEPLEKLPF IWWDEFAD LMMTAGKKIE ELIARLAQKA RAAGIHLILA 751 TQRPSVDVIT GLIKANIPTR IAFQVSSKID SRTILDQMGA ENLLGQGDML 801 FLPPGTAYPQ RVHGAFASDE EVHRWEYLK QFGEPDYVDD ILSGGGSEEL 851 PGIGRSGDGE TDPMYDEAVS WLKTRKASI SGVQRALRIG YNRAARLIDQ 901 MEAEGIVSAP EHNGNRTILV PLDNA* This partial gonococcal sequence contains a predicted transmembrane region and a predicted ATP/GTP-binding site motif A (P-loop; double underlined). Furthermore, it has a domain homologous to the FTSK cell division protein of E. coli. Alignment of ORF58ng and FtsK (accession number p46889) show a 65 % amino acid identity in 459 overlap: ORF58ng: 467 IEEKLAEFKVKVKWDSYSGPVITRYEIEPDVGVRGNSVLNLEKDLARSLGVASIRWET 526 +E +LA+F++K W+ GPVITR+E+ GV+ + NL +DLARSL ++RWE FtsK: 868 VEARLADFRIKADWNYSPGPVITRFELNLAPGVKAARISNLSRDLARSLSTVAVRWEV 927 ORF58ng: 527 IPGKTCMGLELPNPKRQMIRLSEIFNSPEFAESKSKLTLALGQDITGQPWTDLGKAPHL 586 IPGK +GLELPN KRQ + L E+ ++ +F ++ S LT+ LG+DI G+PW DL K PHL FtsK: 928 IPGKPYVGLELPNKKRQTVYLREVLDNAKFRDNPSPLTWLGKDIAGEPWADLAKMPHL 987 ORF58ng: 587 LVAGTTGSGKSVGVNAMILSMLFKAAPEDVRMIMIDPKMLELSIYEGITHLLAPWTDMK 646 LVAGTTGSGKSVGVNAMILSML+KA PEDVR IMIDPKMLELS+YEGI HLL WTDMK FtsK: 988 LVAGTTGSGKSVGVNAMILSMLYKAQPEDVRFIMIDPKMLELSVYEGIPHLLTEWTDMK 1047 ORF58ng: 647 LAANALNWCVNEMEKRYRLMSFMGVRNLAGFNQKIAEAAARGEKIGNPFSLTPDDPEP- 704 AANAL WCVNEME+RY+LMS +GVRNLAG+N+KIAEA I +P+ D + FtsK: 1048 DAANALRWCVNEMERRYKLMSALGVRNLAGYNEKIAEADRMMRPI PDPYWKPGDSMDAQH 1107 ORF58ng: 705 — LEKLPFIWWDEFADLMMTAGKKIEELIARLAQKARAAGIHLILATQRPSVDVITGL 762 L+K P+IW+VDEFADLMMT GKK+EELIARLAQKARAAGIHL+LATQRPSVDVITGL FtsK: 1108 PVLKKEPYIWLVDEFADLMMTVGKKVEELIARLAQKARAAGIHLVLATQRPSVDVITGL 1167 ORF58ng: 763 IKANIPTRIAFQVSSKIDSRTILDQMGAENLLGQGDMLFLPPGTAYPQRVHGAFASDEEV 822 IKANIPTRIAF VSSKIDSRTILDQ GAE+LLG GDML+ P + P RVHGAF D+EV FtsK: 1168 IKANIPTRIAFTVSSKIDSRTILDQAGAESLLGMGDMLYSGPNSTLPVRVHGAFVRDQEV 1227 ORF58ng : 823 HRWEYLKQFGEPDYVDDILSGGGSEELPGIGRSGDGETDPMYDEAVSWLKTRKASISG 882 H W+ K G P YVD I S SE G G G E DP++D+AV V + RKASISG FtsK : 1228 HAVVQDWKARGRPQYVDGITSDSESEGGAG-GFDGAEELDPLFDQAVQFVTEKRKASISG 1286 ORF58ng : 883 VQRALRIGYNRAARLIDQMEAEGIVSAPEHNGNRTILVP 921 VQR RIGYNRAAR+I+QMEA+GIVS HNGNR +L P FtsK : 1287 VQRQFRIGYNRAARIIEQMEAQGIVSEQGHNGNREVLAP 1325 Further work on ORF58ng revealed the complete gonococcal DNA sequence to be <SEQ ID 495>: 1 ATGTTTTGGA TAGTTTTGAT CGTTATtgtg TTGCTTGCGC TTGCCGGCCT 51 GTTTTTTGTC CGCGCACAAT CCGAACGCGA GTGGATGCGC GAGGTTTCTG 101 CGTGGCAGGA AAAGAAAGGG GAAAAACAGG CGGAGCTGCC TGAAATCAAA 151 GACGGTATGC CCGATTTTCC CGAGTTTTCC CTGATGCTTT TCCATGCCGT 201 CAAAACGGCA GTGTATTGGC TGTTTGTCGG TGTCGTCCGT TTCTGCCGAA 251 ACTATCTGGC GCACGAATCC GAACCGGACA GGCCCGTTCC GCCTGCTTCT 301 GCAAACCGTG CGGATGTTCC GACCGCATCC GACGGGTATT CAGACAGTGG 351 AAACGGGACG GAAGAAGCGG AAACGGAAGC AGCAGAAGCT GCGGAGGAAG 401 AGGCTGCCgA TACgGAAGAC ATTGCAACTG CCGTAATCGA CAACCGCCGC 451 ATCCcatTCG ACCGGAGTAT TGCTGAAGGG TTGATGCAGT CTGAAAGCAA 501 AACTTCGCCC GTCCGTCCGG TTTTTAAGGA AATCACTTTG GAAGAAGCAA 551 CGCGTGCTTT AAGCAGCGCG GCTTTAAGGG AAACGAAAAA ACGCTATATC 601 GATGCATTTG AGAAAAACGG AACAGCCGTC CCCAAAGTAC GCGTGTCCGA 651 TACCCCGATG GAAGGGCTGC AGATTATCGG TTTGGACGAC CCTGTGCTTC 701 AACGCACGTA TTCCCGTATG TTTGATGCGG ACAAAGAAGC GTTTTCCGAG 751 TCTGCGGATT ACGGATTTGA GCCGTATTTT GAGAAGCAGC ATCCGTCTGC 801 CTTTTCTGCA GTCAAAGCCG AAAATGCACG GAATGCGCCG TTCCGCCGTC 851 ATGCAGGGCA GGAGAAAGGG CAGGCGGAGG CAAAATCCCC GGATGTTTCC 901 CAAGGGCAGT CCGTTTCAGA CGGCACAGCC GTCCGCGATG CCCGCCGCCG 951 CGTTTCCGTC AATTTGAAAG AACCGAACAA GGCAACGGTT TCTGCGGAGG 1001 CGCGGATTTC GCGCCTGATT CCGGAAAGTC GGACGGTTGT CGGGAAACGG 1051 GATGTCGAAA TGCCGTCTGA AACCGAAAAT GTTTTCACGG AAACCGTTTC 1101 GTCTGTGGGA TACGGCGGTC CGGTTTATGA TGAAGCTGCC GATATCCATA 1151 TTGAAGAGCC TGCCGCGCCC GATGCTTGGG TGGTCGAACC ACCCGAAGTG 1201 CCGGAGGTAG CCGTACCCGA AATCGATATT CTGCCGCCGC CTCCCGTATC 1251 GGAAATCTAC AACCGTACCT ATGAGCCGCC GGCAGGATTC GAGCAGGCGC 1301 AACGCAGCCG CATTGCCGAA ACCGACCATC TTGCCGCTGA TGTTTTGAAT 1351 GGAGGTTGGC AGGAGGAAAC CGCCGCTATT GCAGATGACG GCAGTGAGGG 1401 TGCGGCAGAG CGGTCAAGCG GGCAATATCT GTCGGAAACC GAAGCGTTCG 1451 GGCATGACAG TCAGGCGGTT TGTCCGTTTG AAGATGTGCC GTCTGAACGC 1501 CCGTCCTGCC GGGTATCGGA TACGGAAGCG GATGAAGGGG CGTTCCAATC 1551 GGAAGAGACC GGTGCGGTAT CCGAACACCT GCCGACAACC GACCTGCTTC 1601 TGCCTCCGCT GTTCAATCCC GAGGCGACGC AAACCGAAGA AGAACTGTTG 1651 GAAAACAGCA TCACCATCGA AGAAAAATTG GCGGAGTTCA AAGTCAAGGT 1701 CAAGGTTGTC GATTCTTATT CCGGCCCCGT GATTACGCGT TATGAAATCG 1751 AACCCGATGT CGGCGTGCGC GGCAATTCCG TTCTGAATTT GGAAAAAGAC 1801 TTGGCGCGTT CGCTCGGCGT GGCTTCCATC CGCGTTGTCG AAACCATCCC 1851 CGGCAAAACC TGCATGGGTT TGGAACTTCC GAACCCGAAA CGCCAAATGA 1901 TACGCCTGAG CGAAATTTTC AATTCGCCCG AGTTTGCCGA ATCCAAATCC 1951 AAGCTGACGC TCGCGCTCGG TCAGGACATT ACCGGACAGC CCGTCGTAAC 2001 CGACTTGGGC AAAGCACCGC ATTTGCTGGT TGCCGGCACG ACCGGTTCGG 2051 GCAAATCGGT GGGTGTCAAC GCGATGATTC TGTCTATGCT TTTCAAAGCC 2101 GCGCCGGAAG ACGTGCGTAT GATTATGATC GATCCGAAAA TGCTGGAATT 2151 GAGCATTTAC GAAGGCATCA CGCACCTGCT CGCCCCTGTC GTTACCGATA 2201 TGAAGCTGGC GGCAAACGCG CTGAACTGGT GTGTTAACGA AATGGAAAAA 2251 CGCTACCGCC TGATGAGCTT TATGGGCGTG CGCAATCTTG CGGGCTTCAA 2301 CCAAAAAATC GCCGAAGCCG CAGCAAGGGG AGAAAAAATC GGCAATCCGT 2351 TCAGCCTCAC GCCCGACGAT CCCGAACCTT TGGAAAAACT GCCGTTTATC 2401 GTGGTCGTGG TCGATGAGTT TGCCGATTTG ATGATGACGG CAGGCAAGAA 2451 AATCGAAGAA CTGATTGCGC GCCTCGCCCA AAAAGCCCGC GCGGCAGGCA 2501 TCCACCTTAT CCTTGCCACA CAACGCCCCA GCGTCGATGT CATCACGGGT 2551 CTGATTAAGG CGAACATCCC GACGCGTATC GCGTTCCAAG TGTCCAGCAA 2601 AATCGACAGC CGCACGATTC TCGACCAAAT GGGCGCGGAA AACCTGCTCG 2651 GTCAGGGCGA TATGCTGTTC CTGCCGCCGG GTACTGCCTA TCCGCAGCGC 2701 GTTCACGGCG CGTTTGCCTC GGATGAAGAG GTGCACCGCG TGGTCGAATA 2751 TCTGAAGCAG TTTGGCGAGC CGGACTATGT TGACGATATT TTGAGCGGCG 2801 GCGGCAGCGA AGAGCTGCCC GGCATCGGGC GCAGCGGCGA CGGCGAAACC 2851 GATCCGATGT ACGACGAGGC CGTATCCGTT GTCCTGAAAA CGCGCAAAGC 2901 CAGCATTTCG GGCGTACAGC GCGCCTTGCG CATCGGCTAC AACCGCGCCG 2951 CGCGTCTGAT TGACCAAATG GAAGCGGAAG GCATTGTGTC CGCACCGGAA 3001 CACAACGGCA ACCGTACGAT TCTCGTCCCC TTGGACAATG CTTGA This coπesponds to the amino acid sequence <SEQ ID 496; ORF58ng-l>: 1 MFWIVLIVIV LLALAGLFFV RAQSEREWMR EVSAWQEKKG EKQAELPEIK 51 DGMPDFPEFS LMLFHAVKTA VYWLFVGWR FCRNYLAHES EPDKPVPPAS 101 ANRADVPTAS DGYSDSGNGT EEAETEAAEA AEEEAADTED IATAVIDNRR 151 IPFDRSIAEG LMQSESKTSP VRPVFKEITL EEATRALSSA ALRETKKRYI 201 DAFEKNGTAV PKVRVSDTPM EGLQIIGLDD PVLQRTYSRM FDADKEAFSE 251 SADYGFEPYF EKQHPSAFSA VKAENARNAP FRRHAGQEKG QAEAKSPDVS 301 QGQSVSDGTA VRDARRRVSV NLKEPNKATV SAEARISRLI PESRTWGKR 351 DVEMPSETEN VFTETVSSVG YGGPVYDEAA DIHIEEPAAP DAWWEPPEV 401 PEVAVPEIDI LPPPPVSEIY NRTYEPPAGF EQAQRSRIAE TDHLAADVLN 451 GGWQEETAAI ADDGSEGAAE RSSGQYLSET EAFGHDSQAV CPFEDVPSER 501 PSCRVSDTEA DEGAFQSEET GAVSEHLPTT DLLLPPLFNP EATQTEEELL 551 ENSITIEEKL AEFKVKVKW DΞYSGPVITR YEIEPDVGVR GNSVLNLEKD 601 LARSLGVASI RWETIPGKT CMGLELPNPK RQMIRLSEIF NSPEFAESKS 651 KLTLALGQDI TGQPWTDLG KAPHLLVAGT TGSGKSVGVN AMILSMLFKA 701 APEDVRMIMI DPKMLELSIY EGITHLLAPV VTDMKLAANA LNWCVNEMEK 751 RYRLMSFMGV RNLAGFNQKI AEAAARGEKI GNPFSLTPDD PEPLEKLPFI 801 WWDEFADL MMTAGKKIEE LIARLAQKAR AAGIHLILAT QRPSVDVITG 851 LIKANIPTRI AFQVSSKIDS RTILDQMGAE NLLGQGDMLF LPPGTAYPQR 901 VHGAFASDEE VHRWEYLKQ FGEPDYVDDI LSGGGSEELP GIGRSGDGET 951 DPMYDEAVSV VLKTRKASIS GVQRALRIGY NRAARLIDQM EAEGIVSAPE .001 HNGNRTILVP LDNA* ORF58ng-l and ORF58-1 show 97.2% identity in 1014 aa overlap: 10 20 30 40 50 60 orf58-1. pep MFWIVLIVILLLALAGLFFVRAQSEREWMREVSAWQEKKGEKQAELPEIKDGMPDFPELA II I II II II : I I I I I II I II I II I I II I I II I II I II I I I I I I I I I I I I I I I I II I I I :: orf58ng-l MFWIVLIVIVLLALAGLFFVRAQSEREWMREVSAWQEKKGEKQAELPEIKDGMPDFPEFS 10 20 30 40 50 60 70 80 90 100 110 120 orf58-1. pep LMLFHAVKTAVYWLFVGWRFCRNYLAHESEPDRPVPPASANRADVPTASDGYSDSGNGT II II I I I II I I I I II II I II I II I I II I I I II I II I II I I I II I I I II I I I II I I II II I orf58ng-l LMLFHAVKTAVYWLFVGWRFCRNYLAHESEPDRPVPPASANRADVPTASDGYSDSGNGT 70 80 90 100 110 120 130 140 150 160 170 180 orf58-1. pep EEAETEEAEAAEEEAADTEDIATAVIDNRRIPFDRSIAEGLMPSESEISPVRPVFKEITL I I II I I I I II II II II I II I I II I I I I II I I I I II II II II III: I I II I II I I II I orf58ng-l EEAETEAAEAAEEEAADTEDIATAVIDNRRIPFDRSIAEGLMQSESKTSPVRPVFKEITL 130 140 150 160 170 180 190 200 210 220 230 240 orf58-1. pep EEATRALNSAALRETKKRYIDAFEKNETAVPKVRVSDTPMEGLQIIGLDDPVLQRTYSHM II II I I I : I I I I II I II II I I I II I I I II I II I II I I II II II II I I I I I II I I II I : I orf58ng-l EEATRALSSAALRETKKRYIDAFEKNGTAVPKVRVSDTPMEGLQIIGLDDPVLQRTYSRM 190 200 210 220 230 240 250 260 270 280 290 300 orf58-1. pep FDADKEAFSESADYGFEPYFEKQHPSAFSAVKAENARNAPFHRHAGQGKGQAEAKSPDVS II I I I II I II II II I I I II I I I II I I II II I II I I I I II I I : I I I I I I I II II I I II I I orf58ng-l FDADKEAFSESADYGFEPYFEKQHPSAFSAVKAENARNAPFRRHAGQEKGQAEAKSPDVS 250 260 270 280 290 300 310 320 330 340 350 360 orf58-1. pep QGQSVSDGTAVRDARRRVSVNLKEPNKATVSAEARISRLIPESQTWGKRDVEMPSETEN II I I I II I II II II I II I II I I II I I II II I II I I II I I I I II : I I II I I I I I II I I II I orf58ng-l QGQSVSDGTAVRDARRRVSVNLKEPNKATVSAEARISRLIPESRTWGKRDVEMPSETEN 310 320 330 340 350 360 370 380 390 400 410 420 orf58-1. pep VFTETVSSVGYGGPVYDETADIHIEEPAAPDAWWEPPEVPKVPMTAIDIQPPPPVSEIY II I I I II I II II II I II I: I I I II II I I I II I II I II I I I I: I : III I I II I I II I orf58ng-l VFTETVSSVGYGGPVYDEAADIHIEEPAAPDAWWEPPEVPEVAVPEIDILPPPPVSEIY 370 380 390 400 410 420 430 440 450 460 470 480 orf58-1. pep NRTYEPPSGFEQVQRSRIAETDHLADDVLNGGWQEETAAIADDGSEGAAERSSGQYLSET II II I I I : I II I M II I I I II II I I I I II I II I II I II I I I I I I I II I I II I II I I II I orf58ng-l NRTYEPPAGFEQAQRSRIAETDHLAADVLNGGWQEETAAIADDGSEGAAERSSGQYLSET 430 440 450 460 470 480 490 500 510 520 530 540 orf58-1.pep EAFGHDSQAVCPFENVPSERPSCRVSDTEADEGAFPSEETGAVSEHLPTTDLLLPPLFNP I I II I I I II I II II : II I II I I I II I I II II I I II II I I I I I II I I I I I I II I I I II I I orf58ng-l EAFGHDSQAVCPFEDVPSERPSCRVSDTEADEGAFQSEETGAVSEHLPTTDLLLPPLFNP 490 500 510 520 530 540 550 560 570 580 590 600 orf58-1. pep EATQTEEELLENSITIEEKLAEFKVKVKWDSYSGPVITRYEIEPDVGVRGNSVLNLEKD I I II I I I II I II II I II I I II I I I II I II I II I I II I I I I II II I I I I I I II I II I I II I orf58ng-l EATQTEEELLENSITIEEKLAEFKVKVKWDSYSGPVITRYEIEPDVGVRGNSVLNLEKD 550 560 570 580 590 600 610 620 630 640 650 660 orf58-1. pep LARSLGVASIRWETIPGKTCMGLELPNPKRQMIRLSEIFNSPEFAESKSKLTLALGQDI I I I I I II I II I II I I I I I I I I I I I I I II I I I I II I II I I I I I I II II I I I I II I I I II I I orf58ng-l LARSLGVASIRVVETIPGKTCMGLELPNPKRQMIRLSEIFNSPEFAESKSKLTLALGQDI 610 620 630 640 650 660 670 680 690 700 710 720 orf58-1. pep TGQPWTDLGKAPHLLVAGTTGSGKSVGVNAMILSMLFKAAPEDVRMIMIDPKMLELSIY I I I I I II I II I II I II I II I I I II I I I II II I I II I II II II I I II II I I I I II II I II I orf58ng-l TGQPWTDLGKAPHLLVAGTTGΞGKSVGVNAMILSMLFKAAPEDVRMIMIDPKMLELSIY 670 680 690 700 710 720 730 740 750 760 770 780 orf58-1. pep EGIPHLLAPWTDMKLAANALNWCVNEMEKRYRLMSFMGVRNLAGFNQKIAEAAARGEKI III I II II I I II II I I I II II I II I I I I I I I II I I I II II I I I I I I I I I I I I I I I I I I I orf58ng-l EGITHLLAPWTDMKLAANALNWCVNEMEKRYRLMSFMGVRNLAGFNQKIAEAAARGEKI 730 740 750 760 770 780 7 90 800 810 820 830 840 orf 58-1 . pep GNPFSLTPDDPEPLEKLPFIWWDEFADLMMTAGKKIEELIARLAQKARAAGIHLILAT I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I orf58ng-l GNPFSLTPDDPEPLEKLPFIVWVDEFADLMMTAGKKIEELIARLAQKARAAGIHLILAT 790 800 810 820 830 840 850 860 870 880 890 900 orf58-1. pep QRPSVDVITGLIKANIPTRIAFQVSSKIDSRTILDQMGAENLLGQGDMLFLLPGTAYPQR II I I I II I II I I I I I II II II I II I II I I II II I II I I I I I I I I I I I I I I I II I I II I I orf58ng-l QRPSVDVITGLIKANIPTRIAFQVSSKIDSRTILDQMGAENLLGQGDMLFLPPGTAYPQR 850 860 870 880 890 900 910 920 930 940 950 960 orf58-1. pep VHGAFASDEEVHRWEYLKQFGEPDYVDDILSGGGSEELPGIGRSGDDETDPMYDEAVSV II I II II II I II I I I II II I I I I I I I I I I II I II I II II II II I I II I I I I I I I I I I I I orf58ng-l VHGAFASDEEVHRWEYLKQFGEPDYVDDILSGGGSEELPGIGRSGDGETDPMYDEAVSV 910 920 930 940 950 960 970 980 990 1000 1010 orf58-1.pep VLKTRKASISGVQRALRIGYNRAARLIDQMEAEGIVSAPEHNGNRTILVPLDNAX II I I I I I I II II II I I I I II I I II II I II I I II I II II II I I I I I I II I I II I II orf58ng-l VLKTRKASISGVQRALRIGYNRAARLIDQMEAEGIVSAPEHNGNRTILVPLDNAX 970 980 990 1000 1010 Furthermore, ORF58ng-l shows significant homology to the E.coli protein FtsK: sp|P46889|FTSK_EC0LI CELL DIVISION PROTEIN FTSK >gi | 1651412 | gnl | PID | dl015290 (Dl division protein FtsK [Escherichia coli] >gi | 1651418 | gnl | PID | dl015296 (D90727) Cell division protein FtsK [Escherichia coli] >gi 11787117 (AE000191) cell division protein FtsK [Escherichia coli] Length = 1329 Score = 576 bits (1469), Expect = e-163 Identities = 301/459 (65%), Positives = 353/459 (76%), Gaps = 5/459 (1%) Query: 556 IEEKLAEFKVKVKWDSYSGPVITRYEIEPDVGVRGNSVLNLEKDLARSLGVASIRWET 615 +E +LA+F++K W+ GPVITR+E+ GV+ + NL +DLARSL ++RWE Sbjct: 868 VEARLADFRIKADWNYSPGPVITRFELNLAPGVKAARISNLSRDLARSLSTVAVRWEV 927 Query: 616 IPGKTCMGLELPNPKRQMIRLSEIFNSPEFAESKSKLTLALGQDITGQPWTDLGKAPHL 675 IPGK +GLELPN KRQ + L E+ ++ +F ++ S LT+ LG+DI G+PW DL K PHL Sbjct: 928 IPGKPYVGLELPNKKRQTVYLREVLDNAKFRDNPSPLTWLGKDIAGEPWADLAKMPHL 987 Query: 676 LVAGTTGSGKSVGVNAMILSMLFKAAPEDVRMIMIDPKMLELSIYEGITHLLAPWTDMK 735 LVAGTTGSGKSVGVNAMILSML+KA PEDVR IMIDPKMLELS+YEGI HLL WTDMK Sbjct: 988 LVAGTTGSGKSVGVNAMILSMLYKAQPEDVRFIMIDPKMLELSVYEGIPHLLTEWTDMK 1047 Query: 736 LAANALNWCVNEMEKRYRLMSFMGVRNLAGFNQKIAEAAARGEKIGNPFSLTPDDPEP— 793 AANAL WCVNEME+RY+LMS +GVRNLAG+N+KIAEA I +P+ D + Sbjct: 1048 DAANALRWCVNEMERRYKLMSALGVRNLAGYNEKIAEADRMMRPIPDPYWKPGDSMDAQH 1107 Query: 794 —LEKLPFIWWDEFADLMMTAGKKIEELIARLAQKARAAGIHLILATQRPSVDVITGL 851 L+K P+IW+VDEFADLMMT GKK+EELIARLAQKARAAGIHL+LATQRPSVDVITGL Sbjct: 1108 PVLKKEPYIWLVDEFADLMMTVGKKVEELIARLAQKARAAGIHLVLATQRPSVDVITGL 1167 Query: 852 IKANIPTRIAFQVSSKIDSRTILDQMGAENLLGQGDMLFLPPGTAYPQRVHGAFASDEEV 911 IKANIPTRIAF VSSKIDSRTILDQ GAE+LLG GDML+ P + P RVHGAF D+EV Sbjct: 1168 IKANIPTRIAFTVSSKIDSRTILDQAGAESLLGMGDMLYSGPNSTLPVRVHGAFVRDQEV 1227 Query: 912 HRWEYLKQFGEPDYVDDILSGGGSEELPGIGRSGDGETDPMYDEAVSWLKTRKASISG 971 H W+ K G P YVD I S SE G G G E DP++D+AV V + RKASISG Sbjct: 1228 HAWQDWKARGRPQYVDGITSDSESEGGAG-GFDGAEELDPLFDQAVQFVTEKRKASISG 1286 Query: 972 VQRALRIGYNRAARLIDQMEAEGIVSAPEHNGNRTILVP 1010 VQR RIGYNRAAR+I+QMEA+GIVS HNGNR +L P Sbjct: 1287 VQRQFRIGYNRAARIIEQMEAQGIVSEQGHNGNREVLAP 1325 Based on this analysis, it is predicted that the proteins from N. meningitidis and N.gonorrhoeae, and their epitopes, could be useful antigens for vaccines or diagnostics, or for raising antibodies. Example 59 The following partial DNA sequence was identified in N. meningitidis <SEQ ID 497>: 1 ATGATTTATC AAAGAAACCT CATCAAAGAA CTCTCTTTTA CCGCCGTCGG 51 CATTTTCGTC GTCCTCTTGG CGGTATTGGT CTCCACGCAG GCAATCAACC 101 TGCTCGGCCG TGCCGCCGAC GGGC.GTGA TCGCCATCGA TGCCGTGTTG 151 GCATTGGTCG GCTTCTGGGT C // 901 A TTGCCATCGG TTTGTTTTTA ATTTACCAAA ACGGGCTGAC 951 CCTGCTTTTT GAAGCCGTGG AAGACGGCAA AATCCATTTT TGGCTCGGAC 1001 TGCTGCCTAT GCACATTATC ATGTTTGTCC TTGCACTCAT CCTGTTGCGC 1051 GTCCGCAGTA TGCCCAGCCA GCCCTTCTGG CAGGCGGTTG GCAAAAGTCT 1101 GACATTGAAA GGCGGAAAAT GA This coπesponds to the amino acid sequence <SEQ ID 498; ORF101>: 1 MIYQRNLIKE LSFTAVGIFV VLLAVLVSTQ AINLLGRAAD GXVIAIDAVL 51 ALVGFWV // 301 ...IAIGLFL IYQNGLTLLF EAVEDGKIHF WLGLLPMHII MFVLALILLR 351 VRSMPSQPFW QAVGKSLTLK GGK* Further work revealed the complete nucleotide sequence <SEQ ID 499>: 1 ATGATTTATC AAAGAAACCT CATCAAAGAA CTCTCTTTTA CCGCCGTCGG 51 CATTTTCGTC GTCCTCTTGG CGGTATTGGT CTCCACGCAG GCAATCAACC 101 TGCTCGGCCG TGCCGCCGAC GGGCGTGTCG CCATCGATGC CGTGTTGGCA 151 TTGGTCGGCT TCTGGGTCAT CGGTATGACG CCGCTTTTGC TGGTGTTGAC 201 CGCATTTATC AGTACGTTGA CCGTGTTGAC CCGCTACTGG CGCGACAGCG 251 AAATGTCGGT CTGGCTATCC TGCGGATTGG CATTGAAACA ATGGATACGC 301 CCGGTGATGC AGTTTGCCGT GCCGTTTGCC GTTTTGGTTG CCGTCATGCA 351 GCTTTGGGTG ATACCGTGGG CAGAGCTACG CAGCCGCGAA TACGCTGAAA 401 TCCTGAAGCA GAAGCAGGAA TTGTCTTTGG TGGAGGCAGG CGAGTTCAAC 451 AGTTTGGGCA AGCGCAACGG CAGGGTTTAT TTTGTCGAAA CCTTCGATAC 501 CGAATCCGGC ATCATGAAAA ACCTGTTCCT GCGCGAACAG GACAAAAACG 551 GCGGCGACAA CATCATCTTC GCCAAAGAAG GTAACTTCTC GCTGAACGAC 601 AACAAACGCA CGCTCGAATT GCGCCACGGC TACCGTTACA GCGGCACGCC 651 CGGACGCGCC GACTACAATC AGGTTTCCTT CCAAAAACTC AACCTGATTA 701 TCAGCACCAC GCCCAAACTC ATCGACCCCG TTTCCCACCG CCGTACCATT 751 CCGACCGCCC AACTGATTGG CAGCAGCAAC CCGCAACATC AGGCGGAATT 801 GATGTGGCGC ATCTCGCTGA CCGTCAGCGT CCTCCTACTC TGCCTGCTTG 851 CCGTGCCGCT TTCCTATTTC AACCCGCGCA GCGGACATAC CTACAATATC 901 TTGATTGCCA TCGGTTTGTT TTTAATTTAC CAAAACGGGC TGACCCTGCT 951 TTTTGAAGCC GTGGAAGACG GCAAAATCCA TTTTTGGCTC GGACTGCTGC 1001 CTATGCACAT TATCATGTTT GCCGTTGCAC TCATCCTGTT GCGCGTCCGC 1051 AGTATGCCCA GCCAGCCCTT CTGGCAGGCG GTTGGCAAAA GTCTGACATT 1101 GAAAGGCGGA AAATGA This coπesponds to the amino acid sequence <SEQ ID 500; ORFl 01 -1>: 1 MIYQRNLIKE LSFTAVGIFV VLLAVLVSTQ AINLLGRAAD GRVAIDAVLA 51 LVGFWVIGMT PLLLVLTAFI STLTVLTRYW RDSEMSVWLS CGLALKQWIR 101 PVMQFAVPFA VLVAVMQLWV IPWAELRSRE YAEILKQKQE LSLVEAGEFN 151 SLGKRNGRVY FVETFDTESG IMKNLFLREQ DKNGGDNIIF AKEGNFSLND 201 NKRTLELRHG YRYSGTPGRA DYNQVSFQKL NLIISTTPKL IDPVSHRRTI 251 PTAQLIGSSN PQHQAELMWR ISLTVSVLLL CLLAVPLSYF NPRSGHTYN 301 LIAIGLFLIY QNGLTLLFEA VEDGKIHFWL GLLPMHIIMF AVALILLRVR 351 SMPSQPFWQA VGKSLTLKGG K* Computer analysis of this amino acid sequence gave the following results: Homology with a predicted ORF from N.meninsitidis (sfrain Al ORFlOl shows 91.2% identity over a 57aa overlap and 95.7% identity over a 69aa overlap with an ORF (ORFlOla) from strain A of N meningitidis: 10 20 30 40 50 or 101.pep MIYQRΝLIKELSFTAVGIFWLLAVLVSTQAIΝLLGRAADGXVIAIDAVLALVGFWVX I II II II I I I I I II II I I II II I II I II I II II I I I III I I II I I II II I I I I orf101a MIYQRΝLIKELSFTAVGIFWLLAVLVSTQAIΝLLGXAADXRX-AIDAVLALVGFWVXXM 10 20 30 40 50 // 90 100 110 orf101.pep IAIGLFLIYQΝGLTLLFEAVEDGKIHFWLGL II I II I I I I I I I II II I I I I I II I II II II orf101a LTVSVLLLCLLAVPLSYFΝPRSGHTYΝILXAIGLFLIYQΝGLTLLFEAVEDGKIHFWLGL 280 290 300 310 320 330 120 130 140 150 orf101.pep LPMHIIMFVLALILLRVRSMPSQPFWQAVGKSLTLKGGKX II I I I II I I: M: I I I I II I I II I I II I II II I I II I II I orf101a LPMHIIMFVIAIVLLRVRSMPSQPFWQAVGKSLTLKGGKX 340 350 360 370 The complete length ORFlOla nucleotide sequence <SEQ ID 501> is: 1 ATGATTTATC AAAGAAACCT CATCAAAGAA CTCTCTTTTA CCGCCGTCGG 51 CATTTTCGTC GTCCTCTTGG CGGTATTGGT CTCCACGCAG GCAATCAACC 101 TGCTCGGCCΝ TGCCGCCGAC ΝGGCGTΝTCG CCATCGATGC CGTGTTGGCA 151 TTGGTCGGCT TCTGGGTCΝΝ ΝΝGΝATGACG CCGCTTTTGC TΝGTGTTGAC 201 CGCATTTATC AGTACGTTGA CCGTGTTGAC CCGCTACTGG CGΝGACAGCG 251 AAATGTCGGT CTGGΝTATCC TGCGGATTGG CATTGAAACA ATGGATACGC 301 CCGGTGATGC AGTTTGCCGT GCCGTTTGCC GTTTTGGTTG CCGTCATGCA 351 GCTTTGGGTG ATACCGTGGG CAGAGCTACG CAGCCGCGAA TACGCTGAAA 401 TCCTGAAGCA GAAGCAGGAA TTGTCTTTGG TGGAGGCAGG CGGGTTCAAC 451 AGTTTGGGCA AGCGCAACGG CAGGGTTTAT TTTGTCGAAA CCTTCGATAC 501 CGAATCCGGC ATCATGAAAA ACCTGTTCCT GCGCGAACAG GACAAAAACG 551 GCGGCGACAA CATCATCTTC ΝCCAAAGAAA GTAACTTCTC GCTGAACGAC 601 AACAAACGCA CGCTCGAATT GCGCCACGGC TACCGTTACA GCGGCACGCC 651 CGGACGCGCC GACTACAATC AGGTTTCCTT CCΝAAAACTC AACCTGATTA 701 TCAGCACCAC GCCCAAACTC ATCGACCCCG TTTCCCACCG CCGTACΝATΝ 751 CCΝACΝGCCC AACTGATTGG CAGCAGCAAC CCGCAACATC AΝGCGGAATT 801 GATGTGGCGC ATCTCGCTGA CCGTCAGCGT CCTCCTACTC TGCCTGCTTG 851 CCGTGCCGCT TTCCTATTTC AACCCGCGCA GCGGACATAC CTACAATATC 901 TTGAΝTGCCA TCGGTTTGTT TTTAATTTAC CAAAACGGGC TGACCCTGCT 951 TTTTGAAGCC GTGGAAGACG GCAAAATCCA TTTTTGGCTC GGACTGCTGC 1001 CTATGCACAT CATCATGTTC GTCATCGCAA TCGTACTTCT GCGCGTCCGC 1051 AGCATGCCCA GCCAGCCCTT CTGGCAGGCG GTTGGCAAAA GTCTGACATT 1101 GAAAGGCGGA AAATGA This encodes a protein having amino acid sequence <SEQ LD 502>: 1 MIYQRNLIKE LSFTAVGIFV VLLAVLVSTQ AINLLGXAAD XRXAIDAVLA 51 LVGFWVXXMT PLLLVLTAFI STLTVLTRYW RDSEMSVWXS CGLALKQWIR 101 PVMQFAVPFA VLVAVMQLWV IPWAELRSRE YAEILKQKQE LSLVEAGGFN 151 SLGKRNGRVY FVETFDTESG IMKNLFLREQ DKNGGDNIIF XKESNFSLND 201 NKRTLELRHG YRYSGTPGRA DYNQVSFXKL NLIIΞTTPKL IDPVSHRRTX 251 PTAQLIGSSN PQHXAELMWR ISLTVSVLLL CLLAVPLSYF NPRSGHTYNI_ 301 LXAIGLFLIY QNGLTLLFEA VEDGKIHFWL GLLPMHIIMF VIAIVLLRVR 351 SMPSQPFWQA VGKSLTLKGG K* ORFlOla and ORFlOl-1 show 95.4% identity in 371 aa overlap: orflOla.pep MIYQRNLIKELSFTAVGIFWLLAVLVSTQAINLLGXAADXRXAIDAVLALVGFWVXXMT 60 II I II I II II I I I II I I I II I I II I I II II I I II I I III I I II II I II I II II II orf101-1 MIYQRNLIKELSFTAVGIFWLLAVLVSTQAINLLGRAADGRVAIDAVLALVGFWVIGMT 60 orf101a .pep PLLLVLTAFISTLTVLTRYWRDSEMSVWXSCGLALKQWIRPVMQFAVPFAVLVAVMQLWV 120 I I II I I I I I I I I I I I I I I I I II I I II I I I II I I II I I I I II I I I I I II II II II II II I orf101-1 PLLLVLTAFISTLTVLTRYWRDSEMSVWLSCGLALKQWIRPVMQFAVPFAVLVAVMQLWV 120 orflOla.pep IPWAELRSREYAEILKQKQELSLVEAGGFNSLGKRNGRVYFVETFDTESGIMKNLFLREQ 180 I II II I II I II II II II II II I I I I I I II I I I I I I I I I II I I I II I I I I I I I I I I II I I orf101-1 IPWAELRSREYAEILKQKQELSLVEAGEFNSLGKRNGRVYFVETFDTESGIMKNLFLREQ 180 orflOla.pep DKNGGDNIIFXKESNFSLNDNKRTLELRHGYRYSGTPGRADYNQVSFXKLNLIISTTPKL 240 II II I I I I I I II : I I I I I I I II II II I I II I I I I I I I I I II I I I I I II II II II I I I I orf101-1 DKNGGDNIIFAKEGNFSLNDNKRTLELRHGYRYSGTPGRADYNQVSFQKLNLIISTTPKL 240 orflOla.pep IDPVSHRRTXPTAQLIGSSNPQHXAELMWRISLTVSVLLLCLLAVPLSYFNPRSGHTYNI 300 I I I I II I I I I I I I I I II II I I I II II II II I I I I I I I II II I II I I I II I I I I I I II I orf101-1 IDPVSHRRTIPTAQLIGSSNPQHQAELMWRISLTVSVLLLCLLAVPLSYFNPRSGHTYNI 300 orflOla.pep LXAIGLFLIYQNGLTLLFEAVEDGKIHFWLGLLPMHIIMFVIAIVLLRVRSMPSQPFWQA 360 I I I I I II I II II II I I I I I II I II II II II II I I II I I I :: I :: I II I I I II II II I I I orf101-1 LIAIGLFLIYQNGLTLLFEAVEDGKIHFWLGLLPMHIIMFAVALILLRVRSMPSQPFWQA 360 orflOla.pep VGKSLTLKGGK 371 I I I I I I I I I I I orflOl-1 VGKSLTLKGGK 371 Homology with a predicted ORF from N.sonorrhoeae ORFlOl shows 96.5 % identity in 57aa overlap at the N-terminal domain and 95.1% identity in 6 laa overlap at the C-terminal domain, respectively, with a predicted ORF (ORF 10 lng) from N gonorrhoeae: orf101. pep MIYQRΝLIKELSFTAVGIFWLLAVLVSTQAIΝLLGRAADGXVIAIDAVLALVGFWV 57 II I I I II I II I II I II II II II I I II I I I II II I II I II I I I I I II I I I II I I I I orflOlng MIYQRΝLIKELSFTAVGIF LLAVLVSTQAIΝLLGRAADGRV-AIDAVLALVGFWVIGM 59 // orf101. pep IAIGLFLIYQΝGLTLLFEAVEDGKIHFWLG 333 II I II I I I I I I I II II II I I I II I I I I I I I orflOlng SLTVSVLLLCLLAVPLSYFΝPRSGHTYΝILIAIGLFLIYQΝGLTLLFEAVEDGKIHFWLG 331 orf101. pep LLPMHIIMFVLALILLRVRSMPSQPFWQAVGKSLTLKGGK 373 I I I I I I II I M I :: I II I II II II II I I I II orflOlng LLPMHIIMFVIAIVLLRVRSMPSQPFWQAVG 362 The ORF 10 lng nucleotide sequence <SEQ ED 503> is predicted to encode a protein having partial amino acid sequence <SEQ ED 504>: 1 MIYQRNLIKE LSFTAVGIFV VLLAVLVSTQ AINLLGRAAD GRVAIDAVLA 51 LVGFWVIGMT PLLLVLTAFI STLTVLTRYW RDSEMSVWLS CGLALKQWIR 101 PVMQFAVPFA ILIAVMQLWV IPWAELRSRE YAEILKQKQE LSLVEAGEFN 151 NLGKRNGRVY FVETFDTESG IMKNLFLREQ DKNGGDNIIF AKEGNFSLKD 201 NKRTLELRHG YRYSGTPGRA DYNQVSFQKL NLIISTTPKL IDPVSHRRTI 251 STAQLIGSSN PQHQAELMWR ISLTVSVLLL CLLAVPLSYF NPRSGHTYNI ^ 301 LIAIGLFLIY QNGLTLLFEA VEDGKIHFWL GLLPMHIIMF VIAIVLLRVR 351 SMPSQPFWQA VG... Further work revealed the complete nucleotide sequence <SEQ ID 505>: 1 ATGATTTATC AAAGAAACCT CATCAAAGAA CTCTCTTTTA CCGCCGTCGG 51 CATTTTCGTC GTCCTCTTGG CGGTGTTGGT GTCCACGCAG GCGATCAACC 101 TGCTTGGCCG CGCAGCTGAC GGGCGTGTCG CCATCGATGC CGTGTTGGCC 151 TTAGTCGGCT TCTGGGTCAT CGGTATGACC CCGCTTTTGC TGGTGTTGAC 201 CGCATTCATC AGCACGCTGA CCGTATTGAC CCGCTACTGG CGCGACAGCG 251 AAATGTCGGT CTGGCTATCC TGCGGATTGG CGTTGAAACA GTGGATACGC 301 CCCGTCATGC AGTTTGCCGT GCCGTTTGCC ATCCTGATTG CCGTCATGCA 351 GCTTTGGGTG ATACCGTGGG CAGAGCTGCG CAGCCGCGAA TATGCCGAAA 401 TTTTGAAGCA GAAGCAGGAA TTGTCTTTGG TGGAAGCCGG CGAGTTCAAT 451 AACTTGGGCA AGCGCAACGG CAgggtttaT TtcgtcgaaA CCTTTGACAC 501 CGaatccgGC ATCATGAAAA ACCTGTtcct GcGCGAACAG GACAAAAACG 551 gcggcgacaA CATCATCTTC GCcaaaGAag gtaactTctc gctgaaggaC 601 AACAAAcgca cgctcgaATT GCGCCACGGC TACCGTTACA GCGGcacgcC 651 CGGacGCGCc gactaCAATC AGGTTtcctt cCAAAAacTc aacctgATta 701 TCAGCACCAC GCCCAAacTT ATCGaccCCG TTTCCCACCG CCGCACCATT 751 tcgacCGCCC AAcTGATTGG CAGCAGCAAT CCGCAACATC AGGCAGAATT 801 GATGTGGCGC ATCTCGCTGA CCGTCAGCGT CCTCCTGCTC TGCCTACTCG 851 CCGTGCCGCT TTCCTATTTC AACCCGCGCA GCGGACATAC CTACAATATC 901 TTGATTGCCA TCGGTTTGTT TTTAATTTAC CAAAACGGGC TGACCCTGCT 951 TTTTGAAGCC GTGGAAGACG GCAAAATCCA TTTTTGGCTC GGACTGCTGC 1001 CTATGCACAT CATCATGTTC GTCATCGCAA TCGTACTTCT GCGCGTCCGC 1051 AGTATGCCCA GCCAGCCCTT CTGGCAGGCG GTTGGCAAAA GTCTGACATT 1101 GAAAGgcgGA AAATGA This coπesponds to the .amino acid sequence <SEQ ID 506; ORFlOlng-1 > • 1 MIYQRNLIKE LSFTAVGIFV VLLAVLVSTQ AINLLGRAAD GRVAIDAVLA 51 LVGFWVIGMT PLLLVLTAFI STLTVLTRYW RDSEMSVWLS CGLALKQWIR 101 PVMQFAVPFA ILIAVMQLWV JCPWAELRSRE YAEILKQKQE LSLVEAGEFN 151 NLGKRNGRVY FVETFDTESG IMKNLFLREQ DKNGGDNIIF AKEGNFSLKD 201 NKRTLELRHG YRYSGTPGRA DYNQVSFQKL NLIISTTPKL IDPVSHRRTI 251 STAQLIGSSN PQHQAELMWR ISLTVSVLLL CLLAVPLSYF NPRSGHTYNI 301 LIAIGLFLIY QNGLTLLFEA VEDGKIHFWL GLLPMHIIMF VIAIVLLRVR 351 SMPSQPFWQA VGKSLTLKGG K* ORFlOlng-1 and ORFlOl-1 show 97.6% identity in 371 aa overlap: 10 20 30 40 50 60 orf101-1. pep MIYQRNLIKELSFTAVGIFWLLAVLVSTQAINLLGRAADGRVAIDAVLALVGFWVIGMT II I II I II II II I II II I II I I I I II I I I I I I I I I II I I I I I I I II I I I II I I II I I II I orflOlng-1 MIYQRNLIKELSFTAVGIFWLLAVLVSTQAINLLGRAADGRVAIDAVLALVGFWVIGMT 10 20 30 40 50 60 70 80 90 100 110 120 orf101-1. pep PLLLVLTAFISTLTVLTRYWRDSEMSVWLSCGLALKQWIRPVMQFAVPFAVLVAVMQLWV I II I II I II I I I I I II II I I I II I I I I I I I II II II II II II II I I I II I: I : I I I II I I orflOlng-1 PLLLVLTAFISTLTVLTRYWRDSEMSVWLSCGLALKQWIRPVMQFAVPFAILIAVMQLWV 70 80 90 100 110 120 130 140 150 160 170 180 orf101-1. pep IPWAELRSREYAEILKQKQELSLVEAGEFNSLGKRNGRVYFVETFDTESGIMKNLFLREQ II I I I I II II I II II I I I I I II II I II II I : II I II II I II I I I I I I II I I II I I I II I I orflOlng-1 IPWAELRSREYAEILKQKQELSLVEAGEFNNLGKRNGRVYFVETFDTESGIMKNLFLREQ 130 140 150 160 170 180 190 200 210 220 230 240 orf101-1. pep DKNGGDNIIFAKEGNFSLNDNKRTLELRHGYRYSGTPGRADYNQVSFQKLNLIISTTPKL I I I I I I I I II II I I I II I: II I I I I II I I II II I II I I I I I I I I I I II I I I II I I I II I I orflOlng-1 DKNGGDNIIFAKEGNFSLKDNKRTLELRHGYRYSGTPGRADYNQVSFQKLNLIISTTPKL 190 200 210 220 230 240 250 260 270 280 290 300 orf101-1. pep IDPVSHRRTIPTAQLIGSSNPQHQAELMWRISLTVSVLLLCLLAVPLSYFNPRSGHTYNI I I II I I I I I I I I I I I I I II II I I I II I I I I I I I II II I I I I I I I I II I I I I II I I II I I orflOlng-1 IDPVSHRRTISTAQLIGSSNPQHQAELMWRISLTVSVLLLCLLAVPLSYFNPRSGHTYNI 250 260 270 280 290 300 310 320 330 340 350 360 orf101-1. pep LIAIGLFLIYQNGLTLLFEAVEDGKIHFWLGLLPMHIIMFAVALILLRVRSMPSQPFWQA I II II I I II I I I II I II I II II I I II I II II I I I I I I II I :: M: I I II I I II I I I II I I orflOlng-l LIAIGLFLIYQNGLTLLFEAVEDGKIHFWLGLLPMHIIMFVIAIVLLRVRSMPSQPFWQA 310 320 330 340 350 360 370 orf101-1. pep VGKSLTLKGGKX II I I I II I I I I I orflOlng-l VGKSLTLKGGKX 370 Based on this analysis, including the presence of a putative leader sequence (double-underlined) and several putative transmembrane domains (single-underlined) in the gonococcal protein, it is predicted that the proteins from N. meningitidis and N.gonorrhoeae, and their epitopes, could be useful antigens for vaccines or diagnostics, or for raising antibodies. Example 60 The following partial DNA sequence was identified in N. meningitidis <SEQ ID 507>: 1 .. GGTGGTGGTT TTATCAATGC TTCCTGTGCC ACTTTGACGA CAGCCAAACC 51 GCAATATCAA GCAGGAGACC TTAGCGCTTT TAAGATAAGG CAAGGCAATG 101 TTGTAATCGC CGGACACGGT TTGGATGCAC GTGATACCGA TTACACACGT 151 ATTCTCAGTT ATCATTCCAA AATCGATGCA CCCGTATGGG GACAAGATGT 201 TCGTGTCGTC GCGGGACAAA ACGATGTGGC CGCAACAGGT GATGCACATT 251 CGCCTATTCT CAATAATGCT GCTGCCAATA CGTCAAACAA TACAGCCAAC 301 AACGGCACAC ATATCCCTTT ATTTGCGATT GATACAGGCA AATTAGGAGG 351 TAT.GTATGC CAACAAAATC ACCTTGATCA GTACGGTCGA GCAAGCAGGC 401 ATTCGTAA This coπesponds to the amino acid sequence <SEQ ID 508; ORFl 13>: 1 ..GGGFINASCA TLTTAKPQYQ AGDLSAFKIR QGNWIAGHG LDARDTDYTR 51 ILSYHSKIDA PVWGQDVR AGQNDVAATG DAHSPILNNA AANTSNNTAN 101 NGTHIPLFAI DTGKLGGXVC QQNHLDQYGR ASRHS* Computer analysis of this amino acid sequence gave the following results: Homology with with pspA putative secreted protein of N.meninsitidis (accession AF030941) ORF and pspA show 44% aa identity in 179aa overlap: orf113 GGGFINASCATLTTAKPQYQAGDLSAFKIRQGNWIAGHGLDARDTDYTRILSYHSKIDA 60 GGG INA+ TLT+ P G+L+ F + G WI G GLD D DYTRILS ++I+A pspa GGGLINAASVTLTSGVPVLNNGNLTGFDVSSGKWIGGKGLDTSDADYTRILSRAAEINA 256 orfll3 PVWGQDVRWAGQNDVAATGDAHSPILXXXXXXXXXXXXXXGTHIPLFAIDTGKLGGMYA 120 VWG+DV+W+G+N + G + P AIDT LGGMYA pspa GVWGKDVKWSGKNKLDFDG SLAKTASAPSSSDSVTPTVAIDTATLGGMYA 307 orf113 NKITLISTVEQAGIRNQGQWFASAGNVAVNAEGKLVNTGMIAATGENHAVSLHARNVHN 179 +KITLIST A IRN+G+ FA+ G V ++A+GKL N+G I A +++ A+ V N pspa DKITLISTDNGAVIRNKGRIFAATGGVTLSADGKLSNSGSIDAA EITISAQTVDN 362 Homology with a predicted ORF from Nsonorrhoeae ORFl 13 shows 86.5% identity in 52aa overlap at the N- terminal part and 94.1% identity in 17aa overlap at the C-terminal part with a predicted ORF (ORFl 13ng) from N gonorrhoeae: orf113 GGGFIΝASCATLTTAKPQYQAGDLSAFKIR 30 I I II I I I I I I II I :: I I I I I I I : I : II I I orfll3ng SHPSQLΝGYIEVGGRRAEWIAΝPAGIAVΝGGGFIΝASRATLTTGQPQYQAGDFSGFKIR 224 orfll3 QGΝWIAGHGLDARDTDYTRILSYHSKIDAPVWGQDVRWAGQΝDVAATGDAHSPILΝΝA 90 I I I : II I II II I I II I I : I I I I orfll3ng QGΝAVIAGHGLDARDTDFTRILVCQQΝHLDQYGRTSRHS 263 orfll3 IDTGKLGGXVCQQΝHLDQYGRASRHS 135 I I II I II II II I : II I I orfll3ng DFSGFKIRQGΝAVIAGHGLDARDTDFTRILVCQQΝHLDQYGRTSRHS 263 The complete length ORF113ng nucleotide sequence <SEQ ID 509> is predicted to encode a protein having amino acid sequence <SEQ ID 510>: 1 MNKTLYRVIF NRKRGAWAV AETTKREGKS CADSGSGSVY VKSVSFIPTH 51 SKAFCFSALG FSLCLALGTV NIAFADGIlT DKAAPKTQQA TILQTGNGIP 101 QVNIQTPTSA GVSVNQYAQF DVGNRGAILN NSRSNTQTQL GGWIQGNPWL 151 TRGEARWVN QINSSHPSQL NGYIEVGGRR AEWIANPAG IAVNGGGFIN 201 ASRATLTTGQ PQYQAGDFSG FKIRQGNAVI AGHGLDARDT DFTRILVCQQ 251 NHLDQYGRTS RHS* Based on this analysis, it is predicted that these proteins from N. meningitidis and N.gonorrhoeae, and their epitopes, could be useful antigens for vaccines or diagnostics, or for raising antibodies. Example 61 The following partial DNA sequence was identified in N. meningitidis <SEQ ID 511>: 1 .. TCAACGGGAC ATAGCGAACA AAATTACACT TTGCCGCGAG AAATCACACG 51 CAACATTTCA CTGGGTTCAT TTGCCTATGA ATCGCATCGC AAAGCATTAA 101 GCCATCATGC GCCCAGCCAA GGCACTGAGT TGCCGCAAAG CAACGGTATT 151 TCGCTACCCT ATACGTCCAA TTCTTTTACC CCATTACCCA GCAGCAGCTT 201 ATACATTATC AATCCTGTCA ATAAAGGCTA TCTTGTTGAA ACCGATCCAC 251 GCTTTGCCAA CTACCGTCAA TGGTTGGGTA GTGACTATAT GCtGGACAGC 301 CTCAAACTAG ACCCAAACAA TTTACATAAA CGTTTGGGTG ATGGTTATTA 351 CGAGCAACGT TTAATCAATG AACAAATCGC AGAGCTGACA GGGCATCGTC 401 GTTTAGAcGG TTATCAAAAC GACGAAGAAC AATTTAAAGC CTTAATGGAT 451 AATGGCGCGA CTGCGGCACG TTcGATGAAT CTCAGCGTTG GCATTGCATT 501 AAGTGCCGAG CAAGTAGCGC AACTGACCAG CGATATTGTT TGGTTGGTAC 551 AAAAAGAAGT TAAGCTTCCT GATGGCGGCA CACAAACCGT ATTGGTGCCA 601 CAGGTTTATG TACGCGTTAA AAATGGCGAC ATAGACGGTA AAGGTGCATT 651 GTTGTCAGGC AGCAATACAC AAATCAATGT TTCAGGCAGC CTGAAAAACT 701 CAGGCACGAT TGCAGGgCGC AATGCGCTTA TTATCAATAC CGATACGCTA 751 GACAATATCG GTGGGCGTAT TCATGCGCAA AAATCAGCGG TTACGGCCAC 801 ACAAGACATC AATAATATTG GCGGCATGCT TTCTGCCGAA CAGACATTAT 851 TGCTCAACGC AGGCAACAAC ATCAACAGCC AAAGCACCAC CGCCAGCAGT 901 CAAAATACAC AAGGCAGCAG CACCTACCTA GACCGAATGG CAGGTATTTA 951 TATCACAGGC AAAGAAAAAG GTGTTT .. This coπesponds to the amino acid sequence <SEQ ID 512; ORFl 15>: 1 ..STGHSEQNYT LPREITRNIS LGSFAYESHR KALSHHAPSQ GTELPQSNGI 51 SLPYTSNSFT PLPSSSLYII NPVNKGYLVE TDPRFANYRQ WLGSDYMLDS 101 LKLDPNNLHK RLGDGYYEQR LINEQIAELT GHRRLDGYQN DEEQFKALMD 151 NGATAARSMN LSVGIALSAE QVAQLTSDIV WLVQKEVKLP DGGTQTVLVP 201 QVYVRVKNGD IDGKGALLSG SNTQINVSGS LKNSGTIAGR NALIINTDTL 251 DNIGGRIHAQ KSAVTATQDI NNIGGMLSAE QTLLLNAGNN INSQSTTASS 301 QNTQGSSTYL DRMAGIYITG KEKGV.. Computer analysis of this amino acid sequence gave the following results: Homology with the pspA putative secreted protein of N. meningitidis (accession number AF030941) ORFl 15 and pspA protein show 50% aa identity in 325aa overlap: Orf115: 1 STGHSEQNYTLPREITRNISLGSFAYESHRKALSHHAPSQGTELPQSNGISLPYTSNSFT 60 STG+S Y E++ +1 +G AY+ + + P + NGI +T pspA: 778 STGYSRSPYEPAPEVS-SIRMGISAYKGYAPQQASDIPGTWPWAENGIHPTFT 831 Orfll5: 61 PLPSSSLYIINPVNKGYLVETDPRFANYRQWLGSDYMLDSLKLDPNNLHKRLGDGYYEQR 120 LP+SSL+ I P NKGYL+ETDP F +YR+WLGS YML +L+ DPN++HKRLGDGYYEQ+ pspA: 832 -LPNSSLFAIAPNNKGYLIETDPAFTDYRKWLGSGYMLAALQQDPNHIHKRLGDGYYEQK 890 Orfll5: 121 LINEQIAELTGHRRLDGYQNDEEQFKALMDNGATAARSMNLSVGIALSAEQVAQLTSDIV 180 L+NEQIA+LTG+RRLDGY NDEEQFKALMDNG T A+ + L+ GIALSAEQVA+LTSDIV ppssppAA:: 889911 LVNEQIAKLTGYRRLDGYTNDEEQFKALMDNGITIAKELQLTPGIALSAEQVARLTSDIV 950 Orfll5: 181 WLVQKEVKLPDGGTQTVLVPQVYVRVKNGDIDGKGALLSGSNTQINVSGSLKN-SGTIAG 239 WL + V LPDG TQTVL P+VYVR + D++G+GALLSGS I SG+++N G IAG pspA: 951 WLENETVTLPDGTTQTVLKPKVYVRARPKDMNGQGALLSGSWDIG-SGAIENRGGLIAG 1009 Orfll5: 240 RNALIINTDTLDNIGGRIHAQKSAVTATQDINNIGGMLSAEQTLLLNAGXXXXXXXXXXX 299 R ALI+N + N+ G + + A Dl N G + AE LLL A pspA: 1010 REALILNAQNIKNLQGDLQGKNIFAAAGSDITNTGS-IGAENALLLKASNNIESRSETRS 1068 Orf115: 300 XXXXXXXXXYLDRMAGIYITGKEKG 324 + R+AGIY+TG++ G pspA: 1069 NQNEQGSVRNIGRVAGIYLTGRQNG 1093 Homology with a predicted ORF from N.gonorrhoeae ORFl 15 shows 91.9% identity over a 334aa overlap with a predicted ORF (ORF115ng) from N.gonorrhoeae: orf115. pep STGHSEQNYTLPREITRNISLGSFAYESHRK 31 III I II I I I I : I I I I : I I I I II I I I II I orfllδng NEQTFGEKKVFSENGKLHNYWRARRKGHDETGHREQNYTLPEEITRDISLGSFAYESHSK 71 orf115. pep ALSHHAPSQGTELPQSN GISLPYTSNSFTPLPSSSLYIINPVNKGYLVET 81 I I I: I II I II I I I I I II II II I I I II II I I I : I I II I I I I: II I I I I II orfll5ng ALSRHAPSQGTELPQSNRDNIRTAKSNGISLPYTPNSFTPLPGSSLYIINPANKGYLVET 131 orf115. pep DPRFANYRQWLGSDYMLDSLKLDPNNLHKRLGDGYYEQRLINEQIAELTGHRRLDGYQND 141 II I I I II II I I I I II II I I II I II I I II II I II II I II II I I I I II I I I I II I I I I I II orfll5ng DPRFANYRQWLGSDYMLGSLKLDPNNLHKRLGDGYYEQRLINEQIAELTGHRRLDGYQND 191 orf115. pep EEQFKALMDNGATAARSMNLSVGIALSAEQVAQLTSDIVWLVQKEVKLPDGGTQTVLVPQ 201 II II I I I II II I I II I I II II I I II I II I I : I II II I II II I I II II I I I I II I I II : II orfllδng EEQFKALMDNGATAARSMNLSVGIALSAEQAAQLTSDIVWLVQKEVKLPDGGTQTVLMPQ 251 orf115. pep VYVRVKNGDIDGKGALLSGSNTQINVSGSLKNSGTIAGRNALIINTDTLDNIGGRIHAQK 261 I I I I I I I I I I I II II I I I II I I II I I I I I I II II I II I I II I II II I I I I II I II I I I I orfllδng VYVRVKNGGIDGKGALLSGSNTQINVSGSLKNSGTIAGRNALIINTDTLDNIGGRIHAQK 311 orf115. pep SAVTATQDINNIGGMLSAEQTLLLNAGNNINSQSTTASSQNTQGSSTYLDRMAGIYITGK 321 I I I I I II I II II II : I I I I II I II I I I I I I I : II I : I I I I : I II II II I I II I II II I I orfll 5ng SAVTATQDINNIGGILSAEQTLLLNAGNNINNQSTAKSSQNAQGSSTYLDRMAGIYITGK 371 orf115. pep EKGV 325 II I I orfll5ng EKGVLAAQAGKDINIIAGQISNQSDQGQTRLQAGRDINLDTVQTGKYQEIHFDADNHTIR 431 An ORFl 15ng nucleotide sequence <SEQ ED 513> was predicted to encode a protein having amino acid sequence <SEQ ID 514>: 1 MLVQTEKDGL HNEQTFGEKK VFSENGKLHN YWRARRKGHD ETGHREQNYT 51 LPEEITRDIS LGSFAYESHS KALSRHAPSQ GTELPQSNRD NIRTAKSNGI 101 SLPYTPNSFT PLPGSSLYII NPANKGYLVE TDPRFANYRQ WLGSDYMLGS 151 LKLDPNNLHK RLGDGYYEQR LINEQIAELT GHRRLDGYQN DEEQFKALMD 201 NGATAARSMN LSVGIALSAE QAAQLTSDIV WLVQKEVKLP DGGTQTVLMP 251 QVYVRVKNGG IDGKGALLSG SNTQINVSGS LKNSGTIAGR NALIINTDTL 301 DNIGGRIHAQ KSAVTATQDI NNIGGILSAE QTLLLNAGNN INNQSTAKSS 351 QNAQGSSTYL DRMAGIYITG KEKGVLAAQA GKDINIIAGQ ISNQSDQGQT 401 RLQAGRDINL DTVQTGKYQE IHFDADNHTI RGSTNEVGSS IQTKGDVTLL 451 SGNNLNAKAA EVGSAKGTLA VYAKNDITIS SGIHAGQVDD ASKHTGRSGG 501 GNKLVITDKA QSHHETAQSS TFEGKQWLQ AGNDANILGS NVISDNGTRI 551 QAGNHVRIGT TQTQSQSETY HQTQKSGLMS AGIGFTIGSK TNTQENQSQS 601 NEHTGSTVGS LKGDTTIVAS KHYEQTGSNV SΞPEGNNLIS TQSMDIGAAQ 651 NQLNSKTTQT YEQKGLTVAF SSPVTDLAQQ AIAVAHKAAK QFDKAKTTAL 701 MPWRLPMQVG RLFKQAKAPK K* Further work revealed the following partial gonococcal DNA sequence <SEQ ID 515>: 1 TTGCTTGTGC AAACAGAAAA AGACGGTTTG CATAACGAGC AAACCTTTGG 51 CGAGAAGAAA GTCTTCAGCG AAAATGGTAA GTTGCACAAC TACTGGCGTG 101 CGCGTCGTAA AGGACATGAT GAAACAGGGC ATCGTGAACA AAATTATACT 151 TTGCCGGAGG AAATCACACG CGACATTTCA CTGGGTTCAT TTGCCTATGA 201 ATCGCATAGC AAAGCATTAA GCCGTCATGC GCCCAGCCAA GGCACTGAGT 251 TGCCACAAAG TAACCGGGAT AATATCCGTA CTGCGAAAAG CAACGGTATT 301 TCGCTACCCT ATACGCCCAA TTCTTTTACC CCATTACCCG GCAGCAGCTT 351 ATACATTATC AATCCTGCCA ATAAAGGCTA TCTTGTTGAA ACCGATCCAC 401 GCTTTGCCAA CTACCGTCAA TGGTTGGGTA GTGACTATAT GCTGGGCAGC 451 CTCAAACTAG ACCCAAACAA TTTACATAAA CGTTTGGGTG ATGGTTATTA 501 CGAGCAACGT TTAATCAATG AACAAATCGC AGAGCTGACA GGGCATCGTC 551 GTTTAGACGG TTATCAAAAC GACGAAGAAC AATTTAAAGC CTTAATGGAT 601 AATGGCGCGA CTGCGGCACG TTCGATGAAT CTCAGCGTTG GCATTGCATT 651 AAGTGCCGAG CAAGCAGCGC AACTGACCAG CGATATTGTT TGGTTGGTAC 701 AAAAAGAAGT TAAACTTCCT GATGGCGGCA CACAAACCGT ATTGATGCCA 751 CAGGTTTATG TACGCGTTAA AAATGGCGGC ATAGACGGTA AAGGTGCATT 801 GTTGTCAGGC AGCAATACAC AAATCAATGT TTCAGGCAGC CTGAAAAACT 851 CAGGCACGAT TGCAGGGCGC AATGCGCTTA TTATCAATAC CGATACGCTA 901 GACAATATCG GTGGGCGTAT TCATGCGCAA AAATCAGCGG TTACGGCCAC 951 ACAAGACATC AATAATATTG GCGGCATTCT TTCTGCCGAA CAGACATTAT 1001 TGCTCAATGC GGGTAACAAC ATCAACAACC AAAGCACGGC CAAGAGCAGT 1051 CAAAATGCAC AAGGTAGCAG CACCTACCTA GACCGAATGG CAGGTATTTA 1101 TATCACAGGC AAAGAAAAAG GTGTTTTAGC AGCGCAGGCA GGCAAAGACA 1151 TCAACATCAT TGCCGGTCAA ATCAGCAATC AATCAGATCA AGGGCAAACC 1201 CGGCTGCAGG CAGGACGCGA CATTAACCTG GATACGGTAC AAACCGGCAA 1251 ATATCAAGAA ATCCATTTTG ATGCCGATAA CCATACCATC CGAGGTTCAA 1301 CGAACGAAGT CGGCAGCAGC ATTCAAACAA AAGGCGATGT TACCCtatTG 1351 TCAGGGAATA ATCTCAATGC CAAAGCTGCC GAAGTCGGCA GCGCAAAAGG 1401 CACACTTGCC GTGTATGCTA AAAATGACAT TACTATCAGC TCAGGCATCC 1451 ATGCCGGCCA AGTTGATGAT GCGTCCAAAC ATACAGGCAG AAGCGGCGGC 1501 GGTAATAAAT TAGTCATTAC CGATAAAGCC CAAAGTCATC ACGAAACTGC 1551 TCAAAGCAGC ACCTTTGAAG GCAAGCAAGT TGTATTGCAG GCAGGAAACG 1601 ATGCCAACAT CCTTGGCAGT AATGTTATTT CCGATAATGG CACCCGGATT 1651 CAAGCAGGCA ATCATGTTCG CATTGGTACA ACCCAAACTC AAAGCCAAAG 1701 CGAAACCTAT CATCAAACCC AAAAATCAGG ATTGATGAGT GCAGGTATCG 1751 GCTTCACTAT TGGCAGCAAG ACAAACACAC AAGAAAACCA ATCCCAAAGC 1801 AACGAACATA CAGGCAGTAC CGTAGGCAGC CTGAAAGGCG ATACCACCAT 1851 TGTTGCAAGC AAACACTACG AACAAACCGG CAGCAACGTT TCCAGCCCTG 1901 AGGGCAACAA CCTTATCAGC ACGCAAAGTA TGGATATTGG CGCAGCACAA 1951 AACCAATTAA ACAGCAAAAC CACCCAAACC TACGAACAAA AAGGCTTAAC 2001 GGTGGCATTC AGTTCGCCCG TTACCGATTT GGCACAACAA GCGATTGCCG 2051 TAGCACACAA AGCAGCAAAC AAGTCGGACA AAGCAAAAAC GACCGCGTTA 2101 ATGCCATGGC GGCTGCCAAT GCAGGTTGGC AGGCCTATCA AACAGGCAAA 2151 GGCGCACAAA ACTTAG This coπesponds to the amino acid sequence <SEQ ID 516; ORFl 15ng-l>: 1 LLVQTEKDGL HNEQTFGEKK VFSENGKLHN YWRARRKGHD ETGHREQNYT 51 LPEEITRDIS LGSFAYESHS KALSRHAPSQ GTELPQSNRD NIRTAKSNGI 101 SLPYTPNSFT PLPGSSLYII NPANKGYLVE TDPRFANYRQ WLGSDYMLGS 151 LKLDPNNLHK RLGDGYYEQR LINEQIAELT GHRRLDGYQN DEEQFKALMD 201 NGATAARSMN LSVGIALSAE QAAQLTSDIV WLVQKEVKLP DGGTQTVLMP 251 QVYVRVKNGG IDGKGALLSG SNTQINVSGS LKNSGTIAGR NALIINTDTL 301 DNIGGRIHAQ KSAVTATQDI NNIGGILSAE QTLLLNAGNN INNQSTAKSS 351 QNAQGSSTYL DRMAGIYITG KEKGVLAAQA GKDINIIAGQ ISNQSDQGQT 401 RLQAGRDINL DTVQTGKYQE IHFDADNHTI RGSTNEVGSS IQTKGDVTLL 451 SGNNLNAKAA EVGSAKGTLA VYAKNDITIS SGIHAGQVDD ASKHTGRSGG 501 GNKLVITDKA QSHHETAQSS TFEGKQWLQ AGNDANILGS NVISDNGTRI 551 QAGNHVRIGT TQTQSQSETY HQTQKSGLMS AGIGFTIGSK TNTQENQSQS 601 NEHTGSTVGS LKGDTTIVAS KHYEQTGSNV SSPEGNNLIS TQSMDIGAAQ 651 NQLNSKTTQT YEQKGLTVAF SSPVTDLAQQ AIAVAHKAAN KSDKAKTTAL 701 MPWRLPMQVG RPIKQAKAHK T* This gonococcal protein (ORFl 15ng-l) shows 91.9% identity with ORFl 15 over 334aa: 20 30 40 50 60 70 orfll5ng-l.p NEQTFGEKKVFSENGKLHNYWRARRKGHDETGHREQNYTLPEEITRDISLGSFAYESHSK III I II I I I I : I I I I : I I I I II I M I I I orfll5 STGHSEQNYTLPREITRNISLGSFAYESHRK 10 20 30 80 90 100 110 120 130 orf115ng-l . p ALSRHAPSQGTELPQSNRDNIRTAKSNGISLPYTPNSFTPLPGSSLYIINPANKGYLVET I I I : I I I I I I I II I I I I II I I I II I I I I I MM I I I I I I I: I I I I I I I I orfll5 ALSHHAPSQGTELPQSN GISLPYTSNSFTPLPSSSLYIINPVNKGYLVET 40 50 60 70 80 140 150 160 170 180 190 orfll5ng-l.p DPRFANYRQWLGSDYMLGSLKLDPNNLHKRLGDGYYEQRLINEQIAELTGHRRLDGYQND II I I I II I II II I II I I I II I I I I II II II I II I I I II I I I II II I I I II I II I I I II I orf115 DPRFANYRQWLGSDYMLDSLKLDPNNLHKRLGDGYYEQRLINEQIAELTGHRRLDGYQND 90 100 110 120 130 140 200 210 220 230 240 250 orf115ng-l . p EEQFKALMDNGATAARSMNLSVGIALSAEQAAQLTSDIVWLVQKEVKLPDGGTQTVLMPQ II I II I II I I I I II II I I I I I I I I II I I I I : II I I I I I I I I I II II I I I II I II I I I : II orf115 EEQFKALMDNGATAARSMNLSVGIALSAEQVAQLTSDIVWLVQKEVKLPDGGTQTVLVPQ 150 160 170 180 190 200 260 270 280 290 300 310 orf115ng-l . p VYVRVKNGGIDGKGALLSGSNTQINVSGSLKNSGTIAGRNALIINTDTLDNIGGRIHAQK I II I I I I I I II II I I II II I I I II I II I I II I II I II I I I I I I I II I I I II II I I I II I orfll5 VYVRVKNGDIDGKGALLSGSNTQINVSGSLKNSGTIAGRNALIINTDTLDNIGGRIHAQK 210 220 230 240 250 260 320 330 340 350 360 370 orfll5ng-l.p SAVTATQDINNIGGILSAEQTLLLNAGNNINNQSTAKSSQNAQGSSTYLDRMAGIYITGK II I II I II I II I I I: I I I II I I I II II I I I I : II I : I II I : I I I I I I I II I II I I I I I I orf115 SAVTATQDINNIGGMLSAEQTLLLNAGNNINSQSTTASSQNTQGSSTYLDRMAGIYITGK 270 280 290 300 310 320 380 390 400 410 420 430 orf115ng-l . p EKGVLAAQAGKDINIIAGQISNQSDQGQTRLQAGRDINLDTVQTGKYQEIHFDADNHTIR I I II orfll5 EKGV ition, it shows homology with a secreted N. meningitidis protein in the database: gi 12623258 (AF030941) putative secreted protein [Neisseria meningitidis] Length = 2273 Score = 604 bits (1541), Expect = e-172 Identities = 325/678 (47%), Positives = 449/678 (65%), Gaps = 22/678 (3%) Query: 1 LLVQTEKDGLHNEQTFGEKKVFSENGKLHNYWRARRKGHDETGHREQNYTLPEEITRDIS 60 L+V T + L N++T G K + ++ G LH Y R +KG D TG+ Y E++ I Sbjct: 739 LIVGTPESALDNDETLGTKTI-TDKGDLHRYHRHHKKGRDSTGYSRSPYEPAPEVS-SIR 796 Qι.ery: 61 LGSFAYESHSKALSRHAPSQGTELPQSNRDNIRTAKSNGISLPYTPNSFTPLPGSSLYII 120 +G AY+ + AP Q +++P + + NGI +T LP SSL+ I Sbjct: 797 MGISAYKGY APQQASDIPGTV VPWAENGIHPTFT LPNSSLFAI 840 Query: 121 NPANKGYLVETDPRFANYRQWLGSDYMLGSLKLDPNNLHKRLGDGYYEQRLINEQIAELT 180 P NKGYL+ETDP F +YR+WLGS YML +L+ DPN++HKRLGDGYYEQ+L+NEQIA+LT Sbjct: 841 APNNKGYLIETDPAFTDYRKWLGSGYMLAALQQDPNHIHKRLGDGYYEQKLVNEQIAKLT 900 Query: 181 GHRRLDGYQNDEEQFKALMDNGATAARSMNLSVGIALSAEQAAQLTSDIVWLVQKEVKLP 240 G+RRLDGY NDEEQFKALMDNG T A+ + L+ GIALSAEQ A+LTSDIVWL + V LP Sbjct: 901 GYRRLDGYTNDEEQFKALMDNGITIAKELQLTPGIALSAEQVARLTSDIVWLENETVTLP 960 Query: 241 DGGTQTVLMPQVYVRVKNGGIDGKGALLSGSNTQINVSGSLKN-SGTIAGRNALIINTDT 299 DG TQTVL P+VYVR + ++G+GALLSGS I SG+++N G IAGR ALI+N Sbjct: 961 DGTTQTVLKPKVYVRARPKDMNGQGALLSGSWDIG-SGAIENRGGLIAGREALILNAQN 1019 Query: 300 LDNIGGRIHAQKSAVTATQDINNIGGILSAEQTLLLNAGNNINNQSTAKSSQNAQGSSTY 359 + N+ G + + A Dl N G I AE LLL A NNI ++S +S+QN QGS Sbjct: 1020 IKNLQGDLQGKNIFAAAGSDITNTGSI-GAENALLLKASNNIESRSETRSNQNEQGSVRN 1078 Query: 360 LDRMAGIYITGKEKGVLAAQAGKDINIIAGQISNQSDQGQTRLQAGRDINLDTVQTGKYQ 419 + R+AGIY+TG++ G + AG +1 + A +++NQS+ GQT L AG Dl DT + Q Sbjct: 1079 IGRVAGIYLTGRQNGSVLLDAGNNIVLTASELTNQSEDGQTVLNAGGDIRSDTTGISRNQ 1138 Query: 420 EIHFDADNHTIRGSTNEVGSSIQTKGDVTLLSGNNLNAKAAEVGSAKGTLAVYAKNDITI 479 FD+DN+ IR NEVGS+I+T+G+++L + ++ +AAEVGS +G L + A Dl + Sbjct: 1139 NTIFDSDNYVIRKEQNEVGSTIRTRGNLSLNAKGDIRIRAAEVGSEQGRLKLAAGRDIKV 1198 Query: 480 SSGIHAGQVDDASKHTGRSGGGNKLVITDKAQSHHETAQSSTFEGKQ LQAGNDANILG 539 +G + +DA K+TGRSGGG K +T ++ + A S T +GK+++L +G D + G Sbjct: 1199 EAGKAHTETEDALKYTGRSGGGIKQKMTRHLKNQNGQAVSGTLDGKEIILVSGRDITVTG 1258 Query: 540 SNVISDNGTRIQAGNHVRIGTTQTQSQSETYHQTQKSGLM-SAGIGFTIGSKTNTQENQS 598 SN+I+DN T + A N++ + +T+S+S ++ +KSGLM S GIGFT GSK +TQ N+S Sbjct: 1259 SNIIADNHTILSAKNNIVLKAAETRSRSAEMNKKEKSGLMGSGGIGFTAGSKKDTQTNRS 1318 Query: 599 QSNEHTGSTVGSLKGDTTIVASKHYEQTGSNVSSPEGNNLISTQSMDIGAAQNQLNSKTT 658 ++ HT S VGSL G+T I A KHY QTGS +SSP+G+ IS+ + I AAQN+ + ++ Sbjct: 1319 ETVSHTESWGSLNGNTLISAGKHYTQTGSTISSPQGDVGISSGKISIDAAQNRYSQESK 1378 Query: 659 QTYEQKGLTVAFSSPVTD 676 Q YEQKG+TVA S PV + Sbjct: 1379 QVYEQKGVTVAISVPWN 1396 Based on this analysis, it is predicted that the proteins from N. meningitidis .and N.gonorrhoeae, and their epitopes, could be useful antigens for vaccines or diagnostics, or for raising antibodies. Example 62 The following partial DNA sequence was identified in N. meningitidis <SEQ ID 517>: 1 .. TCAGGGAATA ACCTCAATGC CAAAGCTGCC GAAGTCAGCA GCGCAAACGG 51 TACACTCGCT GTGTCTGCCA ATAATGACAT CAACATCAGC GCAGGCATCA 101 ACACGACCCA TGTTGATGAT GCGTCCAAAC ACACAGGCAG AAGCGGTGGT 151 GGCAATAAAT TAGTCATTAC CGATAAAGCC CAAAGTCATC ACGAAACCGC 201 CCAAAGCAGC ACCTTTGAAG GCAAGCAAGT TGTATTGCAG GCAGGAAACG 251 ATGCCAACAT CCTTGGCAGC AATGTTATTT CCGATAATGG CACCCAGATT 301 CAAGCAGGCA ATCATGTTCG CATTGGTACA ACCCAAACTC AAAGCCAAAG 351 CGAAACCTAT CATCAAACCC AGAAATCAGG ATTGATGAGT GCAGGTATCG 401 GCTTCACTAT TGGCAGCAAG ACAAACACAC AAGAAAACCA ATCCCAAAGC 451 AACGAACATA CAGGCAGTAC CGTAGGCAGC TTGAAAGGCG ATACCACCAT 501 TGTTGCAGGC AAACACTACG AACAAATCGG CAGTACCGTT TCCAGCCCGG 551 AAGGCAACAA TACCATCTAT GCCCAAAGCA TAGACATTCA AGCGGCACAC 601 AACAAATTAA ACAGTAATAC CACCCAAACC TATGAACAAA AAGG.CTAAC 651 GGTGGCATTC AGTTCGCCCG TTACCGATTT GGCACAACAA ... This coπesponds to the .amino acid sequence <SEQ ID 518; ORFl 17>: 1 ..SGNNLNAKAA EVSSANGTLA VSANNDINIS AGINTTHVDD ASKHTGRSGG 51 GNKLVITDKA QSHHETAQSS TFEGKQWLQ AGNDANILGS NVISDNGTQI 101 QAGNHVRIGT TQTQSQSETY HQTQKSGLMS AGIGFTIGSK TNTQENQSQS 151 NEHTGSTVGS LKGDTTIVAG KHYEQIGSTV SSPEGNNTIY AQSIDIQAAH 201 NKLNSNTTQT YEQKXLTVAF SSPVTDLAQQ ... Computer analysis of this amino acid sequence gave the following results: Homology with the pspA putative secreted protein of N. meningitidis (accession number AF030941) ORFl 17 and pspA protein show 45% aa identity in 224aa overlap: Orf117: 4 NLNAKAAEVSSANGTLAVSANNDINISAGINTTHVDDASKHTGRSGGGNKLVITDKAQSH 63 ++ +AAEV S G L ++A Dl + AG T +DA K+TGRSGGG K +T ++ pspA: 1173 DIRIRAAEVGSEQGRLKLAAGRDIKVEAGKAHTETEDALKYTGRSGGGIKQKMTRHLKNQ 1232 Orf117: 64 HETAQSSTFEGKQWLQAGNDANILGSNVISDNGTQIQAGNHVRIGTTQTQSQSETYHQT 123 + A S T +GK+++L +G D + GSN+I+DN T + A N++ + +T+S+S ++ pspA: 1233 NGQAVSGTLDGKEIILVSGRDITVTGSNIIADNHTILSAKNNIVLKAAETRSRSAEMNKK 1292 Orf117: 124 QKSGLM-SAGIGFTIGSKTNTQENQSQSNEHTGSTVGSLKGDTTIVAGKHYEQIGSTVSS 182 +KSGLM S GIGFT GSK +TQ N+S++ HT S VGSL G+T I AGKHY Q GST+SS pspA: 1293 EKSGLMGSGGIGFTAGSKKDTQTNRSETVSHTESWGSLNGNTLISAGKHYTQTGSTIΞS 1352 Orf117: 183 PEGNNTIYAQSIDIQAAHNKLNSNTTQTYEQKXLTVAFSSPVTD 226 P+G+ 1 + I I AA N+ + + Q YEQK +TVA S PV + pspA: 1353 PQGDVGISSGKISIDAAQNRYSQESKQVYEQKGVTVAISVPWN 1396 Homology with a predicted ORF from N.sonorrhoeae ORFl 17 shows 90% identity over a 230aa overlap with a predicted ORF (ORF117ng) from N.gonorrhoeae: orf117.pep SGNNLNAKAAEVSSANGTLAVSANNDINIS 30 I II II I II II I I : II : I I I II I : I II : I I orfll7ng IHFDADNHTIRGSTNEVGSSIQTKGDVTLLSGNNLNAKAAEVGSAKGTLAVYAKNDITIS 480 orf11 .pep AGINTTHVDDASKHTGRSGGGNKLVITDKAQSHHETAQSSTFEGKQWLQAGNDANILGS 90 : \ \ : : : II I I I I II II I I I I I I II I II II II II I I I I I I I I I I I I I I I I I I I I II II I I orfll7ng SGIHAGQVDDASKHTGRSGGGNKLVITDKAQSHHETAQSSTFEGKQWLQAGNDANILGS 540 orf117.pep NVISDNGTQIQAGNHVRIGTTQTQSQSETYHQTQKSGLMSAGIGFTIGSKTNTQENQSQS 150 II II I I I I : I II I I II II II II I I II I I I I I I I II II I I II II I II I II II I II I I I I I I orfll7ng NVISDNGTRIQAGNHVRIGTTQTQSQSETYHQTQKSGLMSAGIGFTIGSKTNTQENQSQS 600 orf117.pep NEHTGSTVGSLKGDTTIVAGKHYEQIGSTVSSPEGNNTIYAQSIDIQAAHNKLNSNTTQT 210 I II I I I I I II I I II I I II I : I II I I I I : I I I I I II I I M M I I : I : I I I : I I II orfll7ng NEHTGSTVGSLKGDTTIVASKHYEQTGSNVSSPEGNNLISTQSMDIGAAQNQLNSKTTQT 660 orf117.pep YEQKXLTVAFSSPVTDLAQQ 230 II II I I I II II II I I I I I I orfll7ng YEQKGLTVAFSSPVTDLAQQAIAVAHKAAKQFDKAKTTALMPWRLPMQVGRLFKQAKAPK 720 An ORFl 17ng nucleotide sequence <SEQ ED 519> was predicted to encode a protein having amino acid sequence <SEQ LD 520>: 1 ..LLVQTEKDGL HNEQTFGEKK VFSENGKLHN YWRARRKGHD ETGHREQNYT 51 LPEEITRDIS LGSFAYESHS KALSRHAPSQ GTELPQSNRD NIRTAKSNGI 101 SLPYTPNSFT PLPGSSLYII NPANKGYLVE TDPRFANYRQ WLGSDYMLGS 151 LKLDPNNLHK RLGDGYYEQR LINEQIAELT GHRRLDGYQN DEEQFKALMD 201 NGATAARSMN LSVGIALSAE QAAQLTSDIV WLVQKEVKLP DGGTQTVLMP 251 QVYVRVKNGG IDGKGALLSG SNTQINVSGS LKNSGTIAGR NALIINTDTL 301 DNIGGRIHAQ KSAVTATQDI NNIGGILSAE QTLLLNAGNN INNQSTAKSS 351 QNAQGSSTYL DRMAGIYITG KEKGVLAAQA GKDINIIAGQ ISNQSDQGQT 401 RLQAGRDINL DTVQTGKYQE IHFDADNHTI RGSTNEVGSS IQTKGDVTLL 451 SGNNLNAKAA EVGSAKGTLA VYAKNDITIS SGIHAGQVDD ASKHTGRSGG 501 GNKLVITDKA QSHHETAQSS TFEGKQWLQ AGNDANILGS NVISDNGTRI 551 QAGNHVRIGT TQTQSQSETY HQTQKSGLMS AGIGFTIGSK TNTQENQSQS 601 NEHTGSTVGS LKGDTTIVAS KHYEQTGSNV SSPEGNNLIS TQSMDIGAAQ 651 NQLNSKTTQT YEQKGLTVAF SSPVTDLAQQ AIAVAHKAAK QFDKAKTTAL 701 MPWRLPMQVG RLFKQAKAPK K* Further work revealed the following gonococcal partial DNA sequence <SEQ ID 521>: 1 TTGCTTGTGC AAACAGAAAA AGACGGTTTG CATAACGAGC AAACCTTTGG 51 CGAGAAGAAA GTCTTCAGCG AAAATGGTAA GTTGCACAAC TACTGGCGTG 101 CGCGTCGTAA AGGACATGAT GAAACAGGGC ATCGTGAACA AAATTATACT 151 TTGCCGGAGG AAATCACACG CGACATTTCA CTGGGTTCAT TTGCCTATGA 201 ATCGCATAGC AAAGCATTAA GCCGTCATGC GCCCAGCCAA GGCACTGAGT 251 TGCCACAAAG TAACCGGGAT AATATCCGTA CTGCGAAAAG CAACGGTATT 301 TCGCTACCCT ATACGCCCAA TTCTTTTACC CCATTACCCG GCAGCAGCTT 351 ATACATTATC AATCCTGCCA ATAAAGGCTA TCTTGTTGAA ACCGATCCAC 401 GCTTTGCCAA CTACCGTCAA TGGTTGGGTA GTGACTATAT GCTGGGCAGC 451 CTCAAACTAG ACCCAAACAA TTTACATAAA CGTTTGGGTG ATGGTTATTA 501 CGAGCAACGT TTAATCAATG AACAAATCGC AGAGCTGACA GGGCATCGTC 551 GTTTAGACGG TTATCAAAAC GACGAAGAAC AATTTAAAGC CTTAATGGAT 601 AATGGCGCGA CTGCGGCACG TTCGATGAAT CTCAGCGTTG GCATTGCATT 651 AAGTGCCGAG CAAGCAGCGC AACTGACCAG CGATATTGTT TGGTTGGTAC 701 AAAAAGAAGT TAAACTTCCT GATGGCGGCA CACAAACCGT ATTGATGCCA 751 CAGGTTTATG TACGCGTTAA AAATGGCGGC ATAGACGGTA AAGGTGCATT 801 GTTGTCAGGC AGCAATACAC AAATCAATGT TTCAGGCAGC CTGAAAAACT 851 CAGGCACGAT TGCAGGGCGC AATGCGCTTA TTATCAATAC CGATACGCTA 901 GACAATATCG GTGGGCGTAT TCATGCGCAA AAATCAGCGG TTACGGCCAC 951 ACAAGACATC AATAATATTG GCGGCATTCT TTCTGCCGAA CAGACATTAT 1001 TGCTCAATGC GGGTAACAAC ATCAACAACC AAAGCACGGC CAAGAGCAGT 1051 CAAAATGCAC AAGGTAGCAG CACCTACCTA GACCGAATGG CAGGTATTTA 1101 TATCACAGGC AAAGAAAAAG GTGTTTTAGC AGCGCAGGCA GGCAAAGACA 1151 TCAACATCAT TGCCGGTCAA ATCAGCAATC AATCAGATCA AGGGCAAACC 1201 CGGCTGCAGG CAGGACGCGA CATTAACCTG GATACGGTAC AAACCGGCAA 1251 ATATCAAGAA ATCCATTTTG ATGCCGATAA CCATACCATC CGAGGTTCAA 1301 CGAACGAAGT CGGCAGCAGC ATTCAAACAA AAGGCGATGT TACCCtatTG 1351 TCAGGGAATA ATCTCAATGC CAAAGCTGCC GAAGTCGGCA GCGCAAAAGG 1401 CACACTTGCC GTGTATGCTA AAAATGACAT TACTATCAGC TCAGGCATCC 1451 ATGCCGGCCA AGTTGATGAT GCGTCCAAAC ATACAGGCAG AAGCGGCGGC 1501 GGTAATAAAT TAGTCATTAC CGATAAAGCC CAAAGTCATC ACGAAACTGC 1551 TCAAAGCAGC ACCTTTGAAG GCAAGCAAGT TGTATTGCAG GCAGGAAACG 1601 ATGCCAACAT CCTTGGCAGT AATGTTATTT CCGATAATGG CACCCGGATT 1651 CAAGCAGGCA ATCATGTTCG CATTGGTACA ACCCAAACTC AAAGCCAAAG 1701 CGAAACCTAT CATCAAACCC AAAAATCAGG ATTGATGAGT GCAGGTATCG 1751 GCTTCACTAT TGGCAGCAAG ACAAACACAC AAGAAAACCA ATCCCAAAGC 1801 AACGAACATA CAGGCAGTAC CGTAGGCAGC CTGAAAGGCG ATACCACCAT 1851 TGTTGCAAGC AAACACTACG AACAAACCGG CAGCAACGTT TCCAGCCCTG 1901 AGGGCAACAA CCTTATCAGC ACGCAAAGTA TGGATATTGG CGCAGCACAA 1951 AACCAATTAA ACAGCAAAAC CACCCAAACC TACGAACAAA AAGGCTTAAC 2001 GGTGGCATTC AGTTCGCCCG TTACCGATTT GGCACAACAA GCGATTGCCG 2051 TAGCACACAA AGCAGCAAAC AAGTCGGACA AAGCAAAAAC GACCGCGTTA 2101 ATGCCATGGC GGCTGCCAAT GCAGGTTGGC AGGCCTATCA AACAGGCAAA 2151 GGCGCACAAA ACTTAG This coπesponds to the amino acid sequence <SEQ ID 522; ORFl 17ng-l>: 1 LLVQTEKDGL HNEQTFGEKK VFSENGKLHN YWRARRKGHD ETGHREQNYT 51 LPEEITRDIS LGSFAYESHS KALSRHAPSQ GTELPQSNRD NIRTAKSNGI 101 SLPYTPNSFT PLPGSSLYII NPANKGYLVE TDPRFANYRQ WLGSDYMLGS 151 LKLDPNNLHK RLGDGYYEQR LINEQIAELT GHRRLDGYQN DEEQFKALMD 201 NGATAARSMN LSVGIALSAE QAAQLTSDIV WLVQKEVKLP DGGTQTVLMP 251 QVYVRVKNGG IDGKGALLSG SNTQINVSGS LKNSGTIAGR NALIINTDTL 301 DNIGGRIHAQ KSAVTATQDI NNIGGILSAE QTLLLNAGNN INNQSTAKSS 351 QNAQGSSTYL DRMAGIYITG KEKGVLAAQA GKDINIIAGQ ISNQSDQGQT 401 RLQAGRDINL DTVQTGKYQE IHFDADNHTI RGSTNEVGSS IQTKGDVTLL 451 SGNNLNAKAA EVGSAKGTLA VYAKNDITIS SGIHAGQVDD ASKHTGRSGG 501 GNKLVITDKA QSHHETAQSS TFEGKQWLQ AGNDANILGS NVISDNGTRI 551 QAGNHVRIGT TQTQSQSETY HQTQKSGLMS AGIGFTIGSK TNTQENQSQS 601 NEHTGSTVGS LKGDTTIVAS KHYEQTGSNV SSPEGNNLIS TQSMDIGAAQ 651 NQLNSKTTQT YEQKGLTVAF SSPVTDLAQQ AIAVAHKAAN KSDKAKTTAL 701 MPWRLPMQVG RPIKQAKAHK T* ORF117ng-l shows the same 90% identity over a 230aa overlap with ORFl 17. In addition, it shows homology with a secreted Nmeningitidis protem in the database: gi I 2623258 (AF030941) putative secreted protein [Neisseria meningitidis] Length = 2273 Score = 604 bits (1541), Expect = e-172 Identities = 325/678 (47%), Positives = 449/678 (65%), Gaps = 22/678 (3%) Query: 1 LLVQTEKDGLHNEQTFGEKKVFSENGKLHNYWRARRKGHDETGHREQNYTLPEEITRDIS 60 L+V T + L N++T G K + ++ G LH Y R +KG D TG+ Y E++ I Sbjct: 739 LIVGTPESALDNDETLGTKTI-TDKGDLHRYHRHHKKGRDSTGYSRSPYEPAPEVS-ΞIR 796 Query: 61 LGSFAYESHSKALSRHAPSQGTELPQSNRDNIRTAKSNGISLPYTPNSFTPLPGSSLYII 120 +G AY+ + AP Q +++P + + NGI +T LP SSL+ I Sbjct: 797 MGISAYKGY APQQASDIPGTV VP AENGIHPTFT LPNSSLFAI 840 Query: 121 NPANKGYLVETDPRFANYRQWLGSDYMLGSLKLDPNNLHKRLGDGYYEQRLINEQIAELT 180 P NKGYL+ETDP F +YR+WLGS YML +L+ DPN++HKRLGDGYYEQ+L+NEQIA+LT Sbjct: 841 APNNKGYLIETDPAFTDYRKWLGSGYMLAALQQDPNHIHKRLGDGYYEQKLVNEQIAKLT 900 Query: 181 GHRRLDGYQNDEEQFKALMDNGATAARSMNLSVGIALSAEQAAQLTSDIVWLVQKEVKLP 240 G+RRLDGY NDEEQFKALMDNG T A+ + L+ GIALSAEQ A+LTSDIVWL + V LP Sbjct: 901 GYRRLDGYTNDEEQFKALMDNGITIAKELQLTPGIALSAEQVARLTSDIVWLENETVTLP 960 Query: 241 DGGTQTVLMPQVYVRVKNGGIDGKGALLSGSNTQINVSGSLKN-SGTIAGRNALIINTDT 299 DG TQTVL P+VYVR + ++G+GALLSGS I SG+++N G IAGR ALI+N Sbjct: 961 DGTTQTVLKPKVYVRARPKDMNGQGALLSGSWDIG-SGAIENRGGLIAGREALILNAQN 1019 Query: 300 LDNIGGRIHAQKSAVTATQDINNIGGILSAEQTLLLNAGNNINNQSTAKSSQNAQGSSTY 359 + N+ G + + A DI N G I AE LLL A NNI ++S +S+QN QGS Sbjct: 1020 IKNLQGDLQGKNIFAAAGSDITNTGSI-GAENALLLKASNNIESRSETRSNQNEQGSVRN 1078 Query: 360 LDRMAGIYITGKEKGVLAAQAGKDINIIAGQISNQSDQGQTRLQAGRDINLDTVQTGKYQ 419 + R+AGIY+TG++ G + AG +1 + A +++NQS+ GQT L AG Dl DT + Q Sbjct: 1079 IGRVAGIYLTGRQNGSVLLDAGNNIVLTASELTNQSEDGQTVLNAGGDIRSDTTGISRNQ 1138 Query: 420 EIHFDADNHTIRGSTNEVGSSIQTKGDVTLLSGNNLNAKAAEVGSAKGTLAVYAKNDITI 479 FD+DN+ IR NEVGS+I+T+G+++L + ++ +AAEVGS +G L + A Dl + Sbjct: 1139 NTIFDSDNYVIRKEQNEVGSTIRTRGNLSLNAKGDIRIRAAEVGSEQGRLKLAAGRDIKV 1198 Query: 480 SSGIHAGQVDDASKHTGRSGGGNKLVITDKAQSHHETAQSSTFEGKQWLQAGNDANILG 539 +G + +DA K+TGRSGGG K +T ++ + A S T +GK+++L +G D + G Sbjct: 1199 EAGKAHTETEDALKYTGRSGGGIKQKMTRHLKNQNGQAVSGTLDGKEIILVSGRDITVTG 1258 Query: 540 SNVISDNGTRIQAGNHVRIGTTQTQSQSETYHQTQKSGLM-SAGIGFTIGSKTNTQENQS 598 SN+I+DN T + A N++ + +T+S+S ++ +KSGLM S GIGFT GSK +TQ N+S Sbjct: 1259 SNIIADNHTILSAKNNIVLKAAETRSRSAEMNKKEKSGLMGSGGIGFTAGSKKDTQTNRS 1318 Query: 599 QSNEHTGSTVGSLKGDTTIVASKHYEQTGSNVSSPEGNNLISTQSMDIGAAQNQLNSKTT 658 ++ HT S VGSL G+T I A KHY QTGS +SSP+G+ IS+ + I AAQN+ + ++ Sbjct: 1319 ETVSHTESWGSLNGNTLISAGKHYTQTGSTISSPQGDVGISSGKISIDAAQNRYSQESK 1378 Query: 659 QTYEQKGLTVAFSSPVTD 676 Q YEQKG+TVA S PV + Sbjct: 1379 QVYEQKGVTVAISVPWN 1396 Based on this analysis, it is predicted that the proteins from N. meningitidis .and N.gonorrhoeae, .and their epitopes, could be useful antigens for vaccines or diagnostics, or for raising antibodies. Example 63 The following partial DNA sequence was identified in N. meningitidis <SEQ ID 523>: 1 ATGATTTACA TCGTACTGTT TCTAGCTGTC GTCCTCGCCG TTGTCGCCTA 51 CAACATGTAT CAGGAAAACC AATACCGCAA AAAAGTGCGC GACCAGTTCG 101 GACACTCCGA CAAAGATGCC CTGCTCAACA GCAwAACCAG CCATGTCCGC 151 GACGGCAAAC CGTCCGGCGG GTCAGTCATG ATGCCGAAAC CCCAACCGGC 201 GGTCAAAAAA ACGGCAAAAC CCCAAGACCC CGyCATGCGC AACCTGCAAG 251 AACAGGATGC CGTCTACATC GCCAAGCAGA AACAGGCAAA AGCCTCCCCG 301 TTCAAAACCG AAATCGAAAC CGCCTTGGAA GAAAGCGGCA TTATCGGCAA 351 CTCCGCCCAC ACCGTTTCCG AACCCCAAAC CGGACATTCC GCAACGAAAC 401 CTGCCGACGC GTCGGCAAAA CCTGCACCCG TTCCGCAAAC ACCTGCAAAA 451 CCGCTGATTA CGCTCAAAGA ACTGTCAAAA GTCGAATTAT CCTGGTTTGA 501 CGTGCGCATC GACTTCATCT CCTAT ... This coπesponds to the .amino acid sequence <SEQ ID 524; ORFl 19>: 1 MIYIVLFLAV VLA AYNMY QENQYRKKVR DQFGHSDKDA LLNSXTSHVR 51 DGKPSGGSVM MPKPQPAVKK TAKPQDPXMR NLQEQDAVYI AKQKQAKASP 101 FKTEIETALE ESGIIGNSAH TVSEPQTGHS ATKPADASAK PAPVPQTPAK 151 PLITLKELSK VELSWFDVRI DFISY... Further work revealed the complete nucleotide sequence <SEQ ID 525>: 1 ATGATTTACA TCGTACTGTT TCTAGCTGTC GTCCTCGCCG TTGTCGCCTA 51 CAACATGTAT CAGGAAAACC AATACCGCAA AAAAGTGCGC GACCAGTTCG 101 GACACTCCGA CAAAGATGCC CTGCTCAACA GCAAAACCAG CCATGTCCGC 151 GACGGCAAAC CGTCCGGCGG GTCAGTCATG ATGCCGAAAC CCCAACCGGC 201 GGTCAAAAAA ACGGCAAAAC CCCAAGACCC CGCCATGCGC AACCTGCAAG 251 AACAGGATGC CGTCTACATC GCCAAGCAGA AACAGGCAAA AGCCTCCCCG 301 TTCAAAACCG AAATCGAAAC CGCCTTGGAA GAAAGCGGCA TTATCGGCAA 351 CTCCGCCCAC ACCGTTTCCG AACCCCAAAC CGGACATTCC GCACCGAAAC 401 CTGCCGACGC GCCGGCAAAA CCTGCACCCG TTCCGCAAAC ACCTGCAAAA 451 CCGCTGATTA CGCTCAAAGA ACTGTCAAAA GTCGAATTAC CCTGGTTTGA 501 CGTGCGCTTC GACTTCATCT CCTATATCGC GCTGACCGAA GCCAAAGAAC 551 TGCACGCACT GCCGCGCCTT TCCAACCGCT GCCGCTACCA GATTGTCGGC 601 TGCACCATGG ACGACCATTT CCAGATTGCC GAACCCATCC CGGGCATCCG 651 CTATCAGGCA TTTATCGTGG GTATTCAGGC AGTCAGCCGC AACGGACTTG 701 CCTCGCAGGA AGAACTCTCC GCATTCAACC GCCAGGTGGA CGCATTCGCA 751 CAAAGCATGG GCGGTCAGAC GCTGCACACC GACCTTGCCG CCTTTATCGA 801 AGTGGCTTCC GCACTGGACG CATTCTGCGC GCGCGTCGAC CAGACCATCG 851 CCATCCATTT GGTTTCCCCG ACCAGCATCA GCGGCGTAGA ACTGCGTTCC 901 GCCGTAACGG GCGTGGGTTT CGTTTTGGAA GACGACGGCG CGTTCCACTA 951 TACCGACACG TCGGGCTCGA CCATGTTCTC CATCTGCTCG CTCAACAACG 1001 AGCCGTTTAC CAACGCCCTT TTGGACAACC AGTCCTACAA AGGCTTCAGT 1051 ATGCTGCTCG ACATCCCGCA CTCTCCGGCA GGCGAAAAAA CCTTCGACGA 1101 TTTGTTTATG GATTTGGCGG TACGCCTGTC CGGCCAGTTG AACCTGAATC 1151 TGGTCAACGA CAAAATGGAA GAAGTTTCGA CCCAATGGCT CAAAGACGTG 1201 CGCACTTATG TATTGGCGCG TCAGTCCGAG ATGCTCAAAG TCGGTATCGA 1251 ACCGGGCGGC AAAACCGCAT TGCGCCTGTT CTCCTAA This coπesponds to the .amino acid sequence <SEQ ID 526; ORFl 19-1 >: 1 MIYIVLFLAV VLAWAYNMY QENQYRKKVR DQFGHSDKDA LLNSKTSHVR 51 DGKPSGGSVM MPKPQPAVKK TAKPQDPAMR NLQEQDAVYI AKQKQAKASP 101 FKTEIETALE ESGIIGNSAH TVSEPQTGHS APKPADAPAK PAPVPQTPAK 151 PLITLKELSK VELPWFDVRF DFISYIALTE AKELHALPRL SNRCRYQIVG 201 CTMDDHFQIA EPIPGIRYQA FIVGIQAVSR NGLASQEELS AFNRQVDAFA 251 QSMGGQTLHT DLAAFIEVAS ALDAFCARVD QTIAIHLVSP TSISGVELRS 301 AVTGVGFVLE DDGAFHYTDT SGSTMFSICS LNNEPFTNAL LDNQSYKGFS 351 MLLDIPHSPA GEKTFDDLFM DLAVRLSGQL NLNLVNDKME EVSTQWLKDV 401 RTYVLARQSE MLKVGIEPGG KTALRLFS* Computer analysis of this amino acid sequence gave the following results: Homology with a predicted ORF from N. meningitidis (sfrain A) ORFl 19 shows 93.7% identity over a 175aa overlap with an ORF (ORFl 19a) from strain A of N meningitidis: 10 20 30 40 50 60 orf 119 . pep MIYIVLFLAWLAWAYΝMYQEΝQYRKKVRDQFGHSDKDALLΝSXTSHVRDGKPSGGSVM I I I I I I I M M I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I orfll 9a MIYIVLFLAAVLAWAYΝMYQEΝQYRKKVRDQFGHSDKDALLΝSKTSHVRDGKPSGGPVM 10 20 30 40 50 60 70 80 90 100 110 120 orf119. pep MPKPQPAVKKTAKPQDPXMRΝLQEQDAVYIAKQKQAKASPFKTEIETALEESGIIGΝSAH II I I II I II I I I I III II I I I I I I I I II II I II II I I I I II I I I II I I I II II II II I orf119a MPKPQPAVKKTAKSQDPAMRΝLQEQDAVYIAKQKQAKASPFKTEIETALEESGIIGΝSAH 70 80 90 100 110 120 130 140 150 160 170 orf119. pep TVSEPQTGHSATKPADASAKPAPVPQTPAKPLITLKELSKVELSWFDVRIDFISY II I I I I I II I I I I I I I I I: I II I I II II I II I I I I I I I I I II I I I : I I I II orfll9a TVPEPQTGHSAPKPADAPAKPVPVPQTPAKPLITLKELSKVELPWFDVRFDFISYIALTE 130 140 150 160 170 180 orf119a AKELHALPRLSΝRCRYQIVGCTMDDHFQIAEPIPGIRYQAFIVGIQAVSRΝGLASQEELS 190 200 210 220 230 240 The complete length ORFl 19a nucleotide sequence <SEQ ID 527> is: 1 ATGATTTACA TCGTACTGTT CCTCGCCGCC GTCCTCGCCG TTGTCGCCTA 51 CAATATGTAT CAGGAAAACC AATACCGCAA AAAAGTGCGC GACCAGTTCG 101 GGCACTCCGA CAAAGATGCC CTGCTCAACA GCAAAACCAG CCATGTCCGC 151 GACGGCAAAC CGTCCGGCGG GCCAGTCATG ATGCCGAAAC CCCAACCGGC 201 GGTCAAAAAA ACGGCAAAAT CCCAAGACCC CGCCATGCGC AACCTGCAAG 251 AGCAGGATGC CGTCTACATC GCCAAGCAGA AACAGGCAAA AGCCTCCCCG 301 TTCAAAACCG AAATCGAAAC CGCCTTGGAA GAAAGCGGCA TTATCGGCAA 351 CTCCGCCCAC ACCGTTCCCG AACCCCAAAC CGGACATTCC GCACCAAAAC 401 CTGCCGACGC GCCGGCAAAA CCTGTTCCCG TTCCGCAAAC GCCGGCAAAA 451 CCGCTGATTA CGCTCAAAGA GCTGTCGAAG GTCGAGCTGC CCTGGTTTGA 501 CGTGCGCTTC GACTTCATCT CTTATATCGC GCTGACCGAA GCCAAAGAAC 551 TGCACGCACT GCCGCGCCTT TCCAACCGCT GCCGCTACCA GATTGTCGGC 601 TGCACCATGG ACGACCATTT CCAGATTGCC GAACCCATCC CGGGCATCCG 651 CTATCAGGCA TTTATCGTGG GTATTCAGGC AGTCAGCCGC AACGGACTTG 701 CCTCGCAGGA AGAACTCTCC GCATTCAACC GCCAGGTGGA TGCATTCGCA 751 CACAGCATGG GCGGTCAGAC GCTGCACACC GACCTTGCCG CCTTTATCGA 801 AGTGGCTTCC GCACTGGACG CATTCTGCGC GCGCGTCGAC CAGACTATCG 851 CCATCCATTT GGTTTCCCCG ACCAGCATCA GCGGCGTAGA ACTGCGTTCC 901 GCCGTAACGG GCGTGGGTTT CGTTTTGGAA GACGACGGCG CGTTCCACTA 951 TACCGACACG TCGGGCTCGA CCATGTTCTC CATCTGCTCG CTCAACAACG 1001 AGCCGTTTAC CAATGCCCTT TTGGACAACC AGTCCTATAA AGGCTTCAGT 1051 ATGCTGCTCG ACATCCCGCA CTCTCCGGCA GGCGAAAAAA CCTTCGACGA 1101 TTTGTTTATG GATTTGGCGG TACGCCTGTC CGGCCAGTTG AACCTGAATC 1151 TGGTCAACGA CAAAATGGAA GAAGTTTCGA CCCAATGGCT CAAAGACGTG 1201 CGCACTTATG TATTGGCTCG TCAGTCCGAG ATGCTCAAAG TCGGTATCGA 1251 ACCGGGCGGC AAAACCGCAT TGCGCCTGTT CTCCTAA This encodes a protein having amino acid sequence <SEQ ID 528>: 1 MIYIVLFLAA VLAWAYNMY QENQYRKKVR DQFGHSDKDA LLNSKTSHVR 51 DGKPSGGPVM MPKPQPAVKK TAKSQDPAMR NLQEQDAVYI AKQKQAKASP 101 FKTEIETALE ESGIIGNSAH TVPEPQTGHS APKPADAPAK PVPVPQTPAK 151 PLITLKELSK VELPWFDVRF DFISYIALTE AKELHALPRL SNRCRYQIVG 201 CTMDDHFQIA EPIPGIRYQA FIVGIQAVSR NGLASQEELS AFNRQVDAFA 251 HSMGGQTLHT DLAAFIEVAS ALDAFCARVD QTIAIHLVSP TSISGVELRS 301 AVTGVGFVLE DDGAFHYTDT SGSTMFSICS LNNEPFTNAL LDNQSYKGFS 351 MLLDIPHSPA GEKTFDDLFM DLAVRLSGQL NLNLVNDKME EVSTQWLKDV 401 RTYVLARQSE MLKVGIEPGG KTALRLFS* ORFl 19a and ORFl 19-1 show 98.6% identity in 428 aa overlap: 10 20 30 40 50 60 orf 119a . pep MIYIVLFLAAVLAWAYNMYQENQYRKKVRDQFGHSDKDALLNSKTSHVRDGKPSGGPVM I I I I I I I I I : I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I orfl l 9-1 MIYIVLFLAWLAWAYNMYQENQYRKKVRDQFGHSDKDALLNSKTSHVRDGKPSGGSVM 10 20 30 40 50 60 70 80 90 100 110 120 orf119a . pep MPKPQPAVKKTAKSQDPAMRNLQEQDAVYIAKQKQAKASPFKTEIETALEESGIIGNSAH I I I I II I II II II I II II I I II I II II I II I I I I I I I II II II II I I I II I I II II I I I orfll9-1 MPKPQPAVKKTAKPQDPAMRNLQEQDAVYIAKQKQAKASPFKTEIETALEESGIIGNSAH 70 80 90 100 110 120 130 140 150 160 170 180 orf119a . pep TVPEPQTGHSAPKPADAPAKPVPVPQTPAKPLITLKELSKVELPWFDVRFDFISYIALTE II I I I II I I II I I II II I I I : I II II I II I I I II I I II I I I I I II I I I II I I II I I I I I orf119-1 TVSEPQTGHSAPKPADAPAKPAPVPQTPAKPLITLKELSKVELPWFDVRFDFISYIALTE 130 140 150 160 170 180 190 200 210 220 230 240 orf119a. pep AKELHALPRLSNRCRYQIVGCTMDDHFQIAEPIPGIRYQAFIVGIQAVSRNGLASQEELS II I I II II I II II II II II I I II I II II I I I I I I II II II II II I I I II I I I I I I II I I I orfll 9-1 AKELHALPRLSNRCRYQIVGCTMDDHFQIAEPIPGIRYQAFIVGIQAVSRNGLASQEELS 190 200 210 220 230 240 250 260 270 280 290 300 orf119 . pep AFNRQVDAFAHSMGGQTLHTDLAAFIEVASALDAFCARVDQTIAIHLVSPTSISGVELRS II I I II I I I I : I II I I I I II I I II I I I II I I I II I I I I I I I I I II I II I I I II II I I I II orfll9-1 AFNRQVDAFAQSMGGQTLHTDLAAFIEVASALDAFCARVDQTIAIHLVSPTSISGVELRS 250 260 270 280 290 300 310 320 330 340 350 360 orf119a. pep AVTGVGFVLEDDGAFHYTDTSGSTMFSICSLNNEPFTNALLDNQSYKGFSMLLDIPHSPA II I I I II I I I I I II II I I I II I I II II I II I I I I I I I I I I I I I I II I I II I I I I I II I I I orfll9-1 AVTGVGFVLEDDGAFHYTDTSGSTMFSICSLNNEPFTNALLDNQSYKGFSMLLDIPHSPA 310 320 330 340 350 360 370 380 390 400 410 420 orf119a. pep GEKTFDDLFMDLAVRLSGQLNLNLVNDKMEEVSTQWLKDVRTYVLARQSEMLKVGIEPGG I II I I I II I II I II I I I I I I II I II II I II I I I I I I II I I II I I II I I II II I I I I II I I orfll 9-1 GEKTFDDLFMDLAVRLSGQLNLNLVNDKMEEVSTQWLKDVRTYVLARQSEMLKVGIEPGG 370 380 390 400 410 420 429 orf ll 9a . pep KTALRLFSX I I I I I I I I I orfll 9-l KTALRLFSX Homology with a predicted ORF from N.sonorrhoeae ORFl 19 shows 93.1% identity over a 175aa overlap with a predicted ORF (ORF119ng) from N.gonorrhoeae: orf 119 . pep MIYIVLFLAWLAWAYNMYQENQYRKKVRDQFGHSDKDALLNSXTSHVRDGKPSGGSVM 60 I I I I I I I I I : I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I orf ll 9ng MIYIVLFLAAVLAWAYNMYQENQYRKKVRDQFGHSDKDALLNSKTSHVRDGKPSGGPVM 60 orf119.pep MPKPQPAVKKTAKPQDPXMRNLQEQDAVYIAKQKQAKASPFKTEIETALEESGIIGNSAH 120 II II I II I I I I I I II I I I II I II I I I I I I I II II I I I I I I I I I I I I I I II I I I I I I orfll9ng MPKPQPAVKKPAKPQDSAMRNLQEQDAVYIAKQKQAKASPFKTEIETALEEIGIIGNSAH 120 orfll9.pep TVSEPQTGHSATKPADASAKPAPVPQTPAKPLITLKELSKVELSWFDVRIDFISY 175 I I I I I I I II I I I I II I I II : I I I I I I I II I I I I I I I I II I I I I I I I : I I I II orfll9ng TVSEPQTGHSAPKPADAPAKPVPVPQTPAKPLITLKELSKVELPWFDVRFDFISYIALTE 180 The coommpplleettee lleenngth ORFl 19ng nucleotide sequence <SEQ ID 529> is: 1 1 4ATGATTTACA TCGTACTGTT CCTCGCCGCC GTCCTCGCCG TTGTCGCCTA 51 ιCAATATGTAT CAGGAAAACC AATACCGCAA AAAAGTGCGC GACCAGTTCG 101 ιGACACTCCGA CAAAGATGCC CTGCTCAACA GCAAAACCAG CCATGTCCGC 151 ιGACGGCAAAC CGTCCGGCGG GCCAGTCATG ATGCCGAAAC CCCAACCGGC 201 ιGGTCAAAAAA CCGGCCAAAC CCCAAGACTC CGCCATGCGC AACCTGCAAG 2 25511 ,AACAGGATGC CGTCTACATC GCCAAGCAGA AACAGGCAAA AGCCTCCCCG 301 'TTCAAAACCG AAATCGAAAC CGCCTTGGAA GAAATCGGCA TTATCGGCAA 351 ιCTCCGCCCAC ACCGTTTCCG AACCCCAAAC CGGACATTCC GCACCGAAAC 401 ιCTGCCGACGC GCCGGCAAAA CCCGTTCCCG TTCCGCAAAC GCCGGCAAAA 451 ( CCGCTGATTA CGCTCAAAGA GCTGTCGAAG GTCGAGCTGC CCTGGTTTGA 5 50011 ιCGTGCGCTtc gACTTCATCT CCTATATCGC GCTGACCGAA GCCAAAGAAC 551 'TGCACGCACT GCCGCGCCTT tccAACCGCT GCCGCTACCA GATTGTCGGC 601 'TGCACCATGG ACGACCATTT CCAGATTGCC GAACCCATCC CGGGCATCCG 651 ιCTATCAGGCA TTTATCGTGG GTATCCAGGC AGTCAGCCGC AACGGACTTG 701 ( CCTCGCAGGA AGAACTCTCC GCATTCAACC GCCAGGCGGA CGCATTCGCA 7 75511 ιCAAAGCATGG GCGGTCAGAC GCTGCACACC GACCTTGCCG CCTTTATCGA 801 ,AGTGGCTTCC GCACTGGACG CATTCTGCGC GCGCGTCGAC CAGACCATCG 851 'CCATCCATTT GGTTTCGCCG ACCAGCATCA GCGGCGTAGA ACTGCGTTCC 901 ( GCCGTAACGG GCGTGGGTTT CGTTTTGGAA GACGACGGCG CGTTCCACTA 951 'TACCGACACG TCGGGCTCGA CCATGTTCTC CATCTGCTCG CTCAACAACG 11000011 ,AGCCGTTTAC CAATGCCCTT TTGGACAACC AGTCCTACAA AGGCTTCAGT 1051 ,ATGCTGCTCG ACATCCCGCA CTCTCCGGCA GGCGAAAAAA CCTTCGACGA 1101 'TTTGTTTATG GATTTGGCGG TACGCCTGTC CGGTCAGTTG AACCTGAATC 1151 'TGGTCAACGA CAAAATGGAA GAAGTTTCGA CCCAATGGCT CAAAGACGTA 1201 *CGCACTTATG TATTGGCGCG TCAGTCCGAG ATGCTCAAAG TCGGTATCGA 11225511 ,ACCGGGCGGC AAAACCGCCC TGCGCCTGTT TTCATAA This encodes a protein having amino acid sequence <SEQ ID 530>: 1 MIYIVLFLAA VLAWAYNMY QENQYRKKVR DQFGHSDKDA LLNSKTSHVR 51 DGKPSGGPVM MPKPQPAVKK PAKPQDSAMR NLQEQDAVYI AKQKQAKASP 101 FKTEIETALE EIGIIGNSAH TVSEPQTGHS APKPADAPAK PVPVPQTPAK 151 PLITLKELSK VELPWFDVRF DFISYIALTE AKELHALPRL SNRCRYQIVG 201 CTMDDHFQIA EPIPGIRYQA FIVGIQAVSR NGLASQEELS AFNRQADAFA 251 QSMGGQTLHT DLAAFIEVAS ALDAFCARVD QTIAIHLVSP TSISGVELRS 301 AVTGVGFVLE DDGAFHYTDT SGSTMFSICS LNNEPFTNAL LDNQSYKGFS 351 MLLDIPHSPA GEKTFDDLFM DLAVRLSGQL NLNLVNDKME EVSTQWLKDV 401 RTYVLARQSE MLKVGIEPGG KTALRLFS* ORFl 19ng and ORFl 19-1 show 98.4% identity over 428 aa overlap: 10 20 30 40 50 60 orfll 9ng MIYIVLFLAAVLAWAYNMYQENQYRKKVRDQFGHSDKDALLNSKTSHVRDGKPSGGPVM I I I I I II I I: II I I II I II I I I I II I II I I I I I I I I I I I I I I I I I I I I II M I I I I I II orfll 9-1 MIYIVLFLAWLAWAYNMYQENQYRKKVRDQFGHSDKDALLNSKTSHVRDGKPSGGSVM 10 20 30 40 50 60 70 80 90 100 110 120 orfll9ng MPKPQPAVKKPAKPQDSAMRNLQEQDAVYIAKQKQAKASPFKTEIETALEEIGIIGNSAH II II II I II I I I I I I I I II I I II I II I I I I II II I I I I I I I II I I I II I II I I I II I orfll9-1 MPKPQPAVKKTAKPQDPAMRNLQEQDAVYIAKQKQAKASPFKTEIETALEESGIIGNSAH 70 80 90 100 110 120 130 140 150 160 170 180 orfll9ng TVSEPQTGHSAPKPADAPAKPVPVPQTPAKPLITLKELSKVELPWFDVRFDFISYIALTE M I I I II I I II II II II II I I : I I II I II I II I II I I I I I I I I I II I I I II I I I I I II I I orfll9-1 TVSEPQTGHSAPKPADAPAKPAPVPQTPAKPLITLKELSKVELPWFDVRFDFISYIALTE 130 140 150 160 170 180 190 200 210 220 230 240 orfll9ng AKELHALPRLSNRCRYQIVGCTMDDHFQIAEPIPGIRYQAFIVGIQAVSRNGLASQEELS I I I I I II I I II I I II I I I II I I II I I I I II I II I I I I I I I I I I II I I II I II I I II I I II orf119-1 AKELHALPRLSNRCRYQIVGCTMDDHFQIAEPIPGIRYQAFIVGIQAVSRNGLASQEELS 190 200 210 220 230 240 250 260 270 280 290 300 orfll9ng AFNRQADAFAQSMGGQTLHTDLAAFIEVASALDAFCARVDQTIAIHLVSPTSISGVELRS I I I II : I I II I I II I I I I I II I I I II I II I II I II II I I II II I I I I I II II I II I I I II orfll9-1 AFNRQVDAFAQSMGGQTLHTDLAAFIEVASALDAFCARVDQTIAIHLVSPTSISGVELRS 250 260 270 280 290 300 310 320 330 340 350 360 orfll9ng AVTGVGFVLEDDGAFHYTDTSGSTMFSICSLNNEPFTNALLDNQSYKGFSMLLDIPHSPA I I I I I I I I I I I I I I I I I II I I II I I II I II I II I I I I I I I I I I I I I I I I I I II I I I II I I orfll9-1 AVTGVGFVLEDDGAFHYTDTSGSTMFSICSLNNEPFTNALLDNQSYKGFSMLLDIPHSPA 310 320 330 340 350 360 370 380 390 400 410 420 orfll9ng GEKTFDDLFMDLAVRLSGQLNLNLVNDKMEEVSTQWLKDVRTYVLARQSEMLKVGIEPGG I I I II I II II II I I I II II I I II II I II I II I I I I I II I I I I I I I I I I II I I I II I I I II orf119-1 GEKTFDDLFMDLAVRLSGQLNLNLVNDKMEEVSTQWLKDVRTYVLARQSEMLKVGIEPGG 370 380 390 400 410 420 429 orfll9ng KTALRLFSX i ii ii i ii i orfll9-l KTALRLFSX Based on this analysis, including the presence of a putative leader sequence in the gonococcal protein, it is predicted that the proteins from N. meningitidis .and N.gonorrhoeae, .and their epitopes, could be useful antigens for vaccines or diagnostics, or for raising antibodies. Example 64 The following partial DNA sequence was identified in N. meningitidis <SEQ ID 531 > 1 ..GCGCGGCACG GCACGGAAGA TTTCTTCATG AACAACAGCG ACAC.ATCAG 51 GCAGATAGTC GAAAGCACCA CCGGTACGAT GAAGCTGCTG ATTTCCTCCA 101 TCGCCCTGAT TTCATTGGTA GTCGGCGGCA TCGGCGTGAT GAACATCATG 151 CTGGTGTCCG TTACCGAGCG CACCAAAGAA ATCGGCATAC GGATGGCAAT 201 CGGCGCGCGG CGCGGCAATA TTTyGCAGCA GTTTTTGATT GAGGCGGTGT 251 TAATCTGCGT CATCGGCGGT TTGGTCGGCG TGGGTTTGTC CGCCGCCGTC 301 AGCCTCGTGT TCAATCATTT TGTAACCGAC TTCCCGATGG ACATTTCCGC 351 CATGTCCGTC ATCGGCGCGG TCGCCTGTTC GACCGGAATC GGCATCGCGT 401 TCGGCTTTAT GCCTGCCAAT AAAGCAGCCA AACTCAATCC GATAGACGCA 451 TTGGCACAGG ATTGA This coπesponds to the amino acid sequence <SEQ ID 532; ORF134>: 1 ..ARHGTEDFFM NNSDXIRQIV ESTTGTMKLL ISSIALISLV VGGIGVMNIM 51 LVSVTERTKE IGIRMAIGAR RGNIXQQFLI EAVLICVIGG LVGVGLSAAV 101 SLVFNHFVTD FPMDISAMSV IGAVACSTGI GIAFGFMPAN KAAKLNPIDA 151 LAQD* Further work revealed the complete nucleotide sequence <SEQ LD 533>: 1 ATGTCGGTGC AAGCAGTATT GGCGCACAAA ATGCGTTCGC TTCTGACGAT 51 GCTCGGCATC ATCATCGGTA TCGCGTCGGT GGTTTCCGTC GTCGCATTGG 101 GCAATGGTTC GCAGAAAAAA ATCCTTGAAG ACATCAGTTC GATAGGGACG 151 AACACCATCA GCATCTTCCC GGGGCGCGGC TTCGGCGACA GGCGCAGCGG 201 CAGGATTAAA ACCCTGACCA TAGACGACGC AAAAATCATC GCCAAACAAA 251 GCTACGTTGC TTCCGCCACG CCCATGACTT CGAGCGGCGG CACGCTGACT 301 TACCGCAACA CCGACCTGAC CGCCTCGCTT TACGGCGTGG GCGAACAATA 351 TTTCGACGTG CGCGGACTGA AGCTGGAAAC GGGGCGGCTG TTTGACGAAA 401 ACGATGTGAA AGAAGACGCG CAGGTCGTCG TCATCGACCA AAATGTCAAA 451 GACAAACTCT TTGCGGACTC GGATCCGTTG GGTAAAACCA TTTTGTTCAG 501 GAAACGCCCC TTGACCGTCA TCGGCGTGAT GAAAAAAGAC GAAAACGCTT 551 TCGGCAATTC CGACGTGCTG ATGCTTTGGT CGCCCTATAC GACGGTGATG 601 CACCAAATCA CAGGCGAGAG CCACACCAAC TCCATCACCG TCAAAATCAA 651 AGACAATGCC AATACCCAGG TTGCCGAAAA AGGGCTGACC GATCTGCTCA 701 AAGCGCGGCA CGGCACGGAA GATTTCTTCA TGAACAACAG CGACAGCATC 751 AGGCAGATAG TCGAAAGCAC CACCGGTACG ATGAAGCTGC TGATTTCCTC 801 CATCGCCCTG ATTTCATTGG TAGTCGGCGG CATCGGCGTG ATGAACATCA 851 TGCTGGTGTC CGTTACCGAG CGCACCAAAG AAATCGGCAT ACGGATGGCA 901 ATCGGCGCGC GGCGCGGCAA TATTTTGCAG CAGTTTTTGA TTGAGGCGGT 951 GTTAATCTGC GTCATCGGCG GTTTGGTCGG CGTGGGTTTG TCCGCCGCCG 1001 TCAGCCTCGT GTTCAATCAT TTTGTAACCG ACTTCCCGAT GGACATTTCC 1051 GCCATGTCCG TCATCGGCGC GGTCGCCTGT TCGACCGGAA TCGGCATCGC 1101 GTTCGGCTTT ATGCCTGCCA ATAAAGCAGC CAAACTCAAT CCGATAGACG 1151 CATTGGCACA GGATTGA This coπesponds to the amino acid sequence <SEQ ID 534; ORF134-l>: 1 MSVQAVLAHK MRSLLTMLGI IIGIASWSV VALGNGSQKK ILEDISSIGT 51 NTISIFPGRG FGDRRSGRIK TLTIDDAKII AKQSYVASAT PMTSSGGTLT 101 YRNTDLTASL YGVGEQYFDV RGLKLETGRL FDENDVKEDA QVWIDQNVK 151 DKLFADSDPL GKTILFRKRP LTVIGVMKKD ENAFGNSDVL MLWSPYTTVM 201 HQITGESHTN SITVKIKDNA NTQVAEKGLT DLLKARHGTE DFFMNNSDSI 251 RQIVESTTGT MKLLISSIAL ISLWGGIGV MNIMLVSVTE RTKEIGIRMA 301 IGARRGNILQ QFLIEAVLIC VIGGLVGVGL SAAVSLVFNH FVTDFPMDIS 351 AMSVIGAVAC STGIGIAFGF MPANKAAKLN PIDALAQD* Computer analysis of this amino acid sequence gave the following results: Homology with the hypothetical protein o648 of E.coli (accession number AE0001891 ORF134 and o648 protein show 45% aa identity in 153aa overlap: Orf134: 2 RHGTEDFFMNNSDXIRQIVESTTGTMKXXXXXXXXXXXWGGIGVMNIMLVSVTERTKEI 61 RHG +DFF N D + + VE TT T++ WGGIGVMNIMLVSVTERT+EI 0648: 496 RHGKKDFFTWNMDGVLKTVEKTTRTLQLFLTLVAVISLWGGIGVMNIMLVSVTERTREI 555 0rfl34: 62 GIRMAIGARRGNIXQQFLIEAXXXXXXXXXXXXXXXXXXXXXFNHFVTDFPMDISAMSVI 121 GIRMA+GAR ++ QQFLIEA F+ + + S ++++ o648: 556 GIRMAVGARASDVLQQFLIEAVLVCLVGGALGITLSLLIAFTLQLFLPGWEIGFSPLALL 615 0rfl34: 122 GAVACSTGIGIAFGFMPANKAAKLNPIDALAQD 154 A CST GI FG++PA AA+L+P+DALA++ o648: 616 LAFLCSTVTGILFGWLPARNAARLDPVDALARE 648 Homology with a predicted ORF from N.meninsitidis (strain Al ORF134 shows 98.7% identity over a 154aa overlap with an ORF (ORF134a) from strain A of N. meningi t idis: 10 20 30 orf 134 . pep ARHGTEDFFMΝΝSDXIRQIVESTTGTMKLL I II I II I I II I I I I I I I I II II I I II I II orf134a GESHTΝSITVKIKDΝAΝTQVAEKGLTDLLKARHGTEDFFMΝΝSDSIRQIVESTTGTMKLL 210 220 230 240 250 260 40 50 60 70 80 90 orfl34.pep ISSIALISLWGGIGVMNIMLVSVTERTKEIGIRMAIGARRGNIXQQFLIEAVLICVIGG I II I I I I I I I II II II II II II I I II I I I II II I II I II II II I I I I I I I II II I I I II orf134a ISSIALISLWGGIGVMNIMLVSVTERTKEIGIRMAIGARRGNILQQFLIEAVLICVIGG 270 280 290 300 310 320 100 110 120 130 140 150 orf134.pep LVGVGLSAAVSLVFNHFVTDFPMDISAMSVIGAVACSTGIGIAFGFMPANKAAKLNPIDA I I I I I II I I II I I I I I I I I I I I I II I II II I II II II I I II I II I II I I I I II II I I I II orf134a LVGVGLSAAVSLVFNHFVTDFPMDISAMSVIGAVACSTGIGIAFGFMPANKAAKLNPIDA 330 340 350 360 370 380 orf134.pep LAQDX I I I II orfl34a LAQDX The complete length ORF134a nucleotide sequence <SEQ ID 535> is: 1 ATGTCGGTGC AAGCAGTATT GGCGCACAAA ATGCGTTCGC TTCTGACGAT 51 GCTCGGCATC ATCATCGGTA TCGCTTCGGT TGTCTCCGTC GTCGCATTGG 101 GCAACGGTTC GCAGAAAAAA ATCCTTGAAG ACATCAGTTC GATAGGGACG 151 AACACCATCA GCATCTTCCC AGGGCGCGGC TTCGGCGACA GGCGCAGCGG 201 CAGGATTAAA ACCCTGACCA TAGACGACGC AAAAATCATC GCCAAACAAA 251 GCTACGTTGC TTCCGCCACG CCCATGACTT CGAGCGGCGG CACGCTGACT 301 TACCGCAATA CCGACCTGAC CGCTTCTTTG TACGGTGTGG GCGAACAATA 351 TTTCGACGTG CGCGGGCTGA AGCTGGAAAC GGGGCGGCTG TTTGACGAAA 401 ACGATGTGAA AGAAGACGCG CAGGTCGTCG TCATCGACCA AAATGTCAAA 451 GACAAACTCT TTGCGGACTC GGATCCGTTG GGTAAAACCA TTTTGTTCAG 501 GAAACGCCCC TTGACCGTCA TCGGCGTGAT GAAAAAAGAC GAAAACGCTT 551 TCGGCAATTC CGACGTGCTG ATGCTTTGGT CGCCCTATAC GACGGTGATG 601 CACCAAATCA CAGGCGAGAG CCACACCAAC TCCATCACCG TCAAAATCAA 651 AGACAATGCC AATACCCAGG TTGCCGAAAA AGGGCTGACC GATCTGCTCA 701 AAGCGCGGCA CGGCACGGAA GATTTCTTCA TGAACAACAG CGACAGCATC 751 AGGCAGATAG TCGAAAGCAC CACCGGTACG ATGAAGCTGC TGATTTCCTC 801 CATCGCCCTG ATTTCATTGG TAGTCGGCGG CATCGGCGTG ATGAACATCA 851 TGCTGGTGTC CGTTACCGAG CGCACCAAAG AAATCGGCAT ACGGATGGCA 901 ATCGGCGCGC GGCGCGGCAA TATTTTGCAG CAGTTTTTGA TTGAGGCGGT 951 GTTAATCTGC GTCATCGGCG GTTTGGTCGG CGTGGGTTTG TCCGCCGCCG 1001 TCAGCCTCGT GTTCAATCAT TTTGTAACCG ACTTCCCGAT GGACATTTCC 1051 GCCATGTCCG TCATCGGCGC GGTCGCCTGT TCGACCGGAA TCGGCATCGC 1101 GTTCGGCTTT ATGCCTGCCA ATAAAGCAGC CAAACTCAAT CCGATAGATG 1151 CATTGGCGCA GGATTGA This encodes a protein having amino acid sequence <SEQ ID 536>: 1 MSVQAVLAHK MRSLLTMLGI IIGIASWSV VALGNGSQKK ILEDISSIGT 51 NTISIFPGRG FGDRRSGRIK TLTIDDAKII AKQSYVASAT PMTSSGGTLT 101 YRNTDLTASL YGVGEQYFDV RGLKLETGRL FDENDVKEDA QVWIDQNVK 151 DKLFADSDPL GKTILFRKRP LTVIGVMKKD ENAFGNSDVL MLWSPYTTVM 201 HQITGESHTN SITVKIKDNA NTQVAEKGLT DLLKARHGTE DFFMNNSDSI 251 RQIVESTTGT MKLLISSIAL ISLWGGIGV MNIMLVSVTE RTKEIGIRMA 301 IGARRGNILQ QFLIEAVLIC VIGGLVGVGL SAAVSLVFNH FVTDFPMDIS 351 AMSVIGAVAC STGIGIAFGF MPANKAAKLN PIDALAQD* ORF134a and ORF134-1 show 100.0% identity in 388 aa overlap: orf134a . pep MSVQAVLAHKMRSLLTMLGIIIGIASWSWALGNGSQKKILEDISSIGTNTISIFPGRG I I I I I I I I I I I I II I I I I I I I I I I I I II II I I II I I I I II I I I II I II I I I I I II II I I I orf134-1 MSVQAVLAHKMRSLLTMLGIIIGIASWSWALGNGSQKKILEDISSIGTNTISIFPGRG orf134a . pep FGDRRSGRIKTLTIDDAKIIAKQSYVASATPMTSSGGTLTYRNTDLTASLYGVGEQYFDV I I II I I I I I I I I I I II I I I I I II I I II I I I I I I I I II I I I I II I I I I I II I I II II I I II orf134-1 FGDRRSGRIKTLTIDDAKIIAKQSYVASATPMTSSGGTLTYRNTDLTASLYGVGEQYFDV orf134a.pep RGLKLETGRLFDENDVKEDAQVWIDQNVKDKLFADSDPLGKTILFRKRPLTVIGVMKKD I I I I I I I I I I I II II I I I II M I I II II I II I I I I I I I I I I I I I II I I I I I II I I I I II I orf134-1 RGLKLETGRLFDENDVKEDAQVWIDQNVKDKLFADSDPLGKTILFRKRPLTVIGVMKKD orf134a . pep ENAFGNSDVLMLWSPYTTVMHQITGESHTNSITVKIKDNANTQVAEKGLTDLLKARHGTE I III I I I I I I I I II I II I I I I I I II I II I II I I I II I I I I II II I I I I I II II I I I I II I orf134-1 ENAFGNSDVLMLWSPYTTVMHQITGESHTNSITVKIKDNANTQVAEKGLTDLLKARHGTE orf134a . pep DFFMNNSDSIRQIVESTTGTMKLLISSIALISLWGGIGVMNIMLVSVTERTKEIGIRMA I I I I II I II I II I I I II II II I II I II II II I I II I I II I I I I I I I I I I II I I I II I I II orf134-1 DFFMNNSDSIRQIVESTTGTMKLLISSIALISLWGGIGVMNIMLVSVTERTKEIGIRMA orf134a . pep IGARRGNILQQFLIEAVLICVIGGLVGVGLSAAVSLVFNHFVTDFPMDISAMSVIGAVAC I I II II I II I I I II I I I I I I I I I II II I I I II I I I I II II II II I II I I I II I I I I I II I orf134-1 IGARRGNILQQFLIEAVLICVIGGLVGVGLSAAVSLVFNHFVTDFPMDISAMSVIGAVAC orf134a.pep STGIGIAFGFMPANKAAKLNPIDALAQDX II I II I I I I II I I I I I I I II I II I I I II I orf134-1 STGIGIAFGFMPANKAAKLNPIDALAQDX Homology with a predicted ORF from N.gonorrhoeae ORF134 shows 96.8% identity over a 154aa overlap with a predicted ORF (ORF134.ng) from N. gonorrhoeae: orf134. pep ARHGTEDFFMNNSDXIRQIVESTTGTMKLL 30 I I I II I I I I I I I I I II IM II II I I I I I I orfl34ng GESHTNSITVKIKDNANTRVAEKGLAELLKARHGTEDFFMNNSDSIRQMVESTTGTMKLL 264 orf134.pep ISSIALISLWGGIGVMNIMLVSVTERTKEIGIRMAIGARRGNIXQQFLIEAVLICVIGG 90 I I I I I I II I I I I I I II I I II I I II I I I I I II II I I I I I II I I II I I I I I I II I I I : II I orfl34ng ISSIALISLWGGIGVMNIMLVSVTERTKEIGIRMAIGARRGNILQQFLIEAVLICIIGG 324 orfl34.pep LVGVGLSAAVSLVFNHFVTDFPMDISAMSVIGAVACSTGIGIAFGFMPANKAAKLNPIDA 150 I I I I I I II I II II I I I II I I I I I I I II I I I I II II I I I I I II I I II I I II I II I I I I I I orfl34ng LVGVGLSAAVSLVFNHFVTDFPMDISAASVIGAVACSTGIGIAFGFMPANKAAKLNPIDA 384 orfl34.pep LAQD 154 M M orfl34ng LAQD 388 The complete length ORF134ng nucleotide sequence <SEQ D 537> is: 1 ATGTCGGTGC AAGCAGTATT GGCGCACAAA ATGCGTTCGC TTCTGACCAT 51 GCTCGGCATC ATCATCGGTA TCGCTTCGGT TGTCTCCGTC GTCGCGCTGG 101 GCAACGGTTC GCAGAAAAAA ATCCTCGAAG ACATCAGTTC GATGGGGACG 151 AACACCATCA GCATCTTCCC CGGGCGCGGC TTCGGCGACA GGCGCAGCGG 201 CAAAATCAAA ACCCTGACCA TAGACGACGC AAAAATCATC GCCAAACAAA 251 GCTACGTTGC CTCCGCCACG CCCATGACTT CGAGCGGCGG CACGCTGACC 301 TACCGCAATA CCGACCTGAC CGCTTCTTTG TACGGTGTGG GCGAACAATA 351 TTTCGACGTG CGCGGGCTGA AGCTGGAAAC GGGGCGGCTG TTTGATGAGA 401 ACGATGTGAA AGAAGACGCG CAAGTCGTCG TCATCGACCA AAATGTCAAA 451 GACAAACTCT TTGCGGACTC GGATCCGTTG GGTAAAACCA TTTTGTTCAG 501 GAAACGCCCC TTGACCGTCA TCGGCGTGAT GAAAAAAGAC GAAAACGCTT 551 TCGGCAATTC CGACGTGCTG ATGCTTTGGT CGCCCTATAC GACGGTGATG 601 CACCAAATCA CAGGCGAGAG CCACACCAAC TCCATCACCG TCAAAATCAA 651 AGACAATGCC AATACCCGGG TTGCCGAAAA AGGGCTGGCC GAGCTGCTCA 701 AAGCACGGCA CGGCACGGAA GACTTCTTTA TGAACAACAG CGACAGCATC 751 AGGCAGATGG TCGAAAGCAC CACCGGTACG ATGAAGCTGC TGATTTCCTC 801 CATCGCCCTG ATTTCATTGG TAGTCGGCGG CATCGGTGTG ATGAACATTA 851 TGCTGGTGTC CGTTACCGAG CGCACCAAAG AAATCGGCAT ACGGATGGCA 901 ATCGGCGCGC GGCGCGGCAA TATTTTGCAG CAGTTTTTGA TTGAGGCGGT 951 GTTAATCTGC ATCATCGGAG GCTTGGTCGG CGTAGGTTTG TCCGCCGCCG 1001 TCAGCCTCGT GTTCAATCAT TTTGTAACCG ATTTCCCGAT GGACATTTCG 1051 GCGGCATCCG TTATCGGGGC GGTCGCCTGT TCGACCGGAA TCGGCATCGC 1101 GTTCGGCTTT ATGCCTGCCA ATAAGGCAGC CAAACTCAAT CCGATAGATG 1151 CATTGGCGCA GGATTGA This encodes a protein having amino acid sequence <SEQ LD 538>: 1 MSVQAVLAHK MRSLLTMLGI IIGIASWSV VALGNGSQKK ILEDISSMGT 51 NTISIFPGRG FGDRRSGKIK TLTIDDAKII AKQSYVASAT PMTSSGGTLT 101 YRNTDLTASL YGVGEQYFDV RGLKLETGRL FDENDVKEDA QVWIDQNVK 151 DKLFADSDPL GKTILFRKRP LTVIGVMKKD ENAFGNSDVL MLWSPYTTVM 201 HQITGESHTN SITVKIKDNA NTRVAEKGLA ELLKARHGTE DFFMNNSDSI 251 RQMVESTTGT MKLLISSIAL ISLWGGIGV MNIMLVSVTE RTKEIGIRMA 301 IGARRGNILQ QFLIEAVLIC IIGGLVGVGL SAAVSLVFNH FVTDFPMDIS 351 AASVIGAVAC STGIGIAFGF MPANKAAKLN PIDALAQD* ORF134ng and ORF134-1 show 97.9% identity in 388 aa overlap: orfl34ng MSVQAVLAHKMRSLLTMLGIIIGIASWSWALGNGSQKKILEDISSMGTNTISIFPGRG I I I I I II I I I I I I I II I I II I II I I I I I I I II II II I II I I II I I II : I II I II I I I I I I orf134-1 MSVQAVLAHKMRSLLTMLGIIIGIASWSWALGNGSQKKILEDISSIGTNTISIFPGRG orfl34ng FGDRRSGKIKTLTIDDAKIIAKQSYVASATPMTSSGGTLTYRNTDLTASLYGVGEQYFDV II I I I II : I II I I I I I I I II I II I I I I I I I I I II II I II I I I I I I I I I II I I I II I II I I orf134-1 FGDRRSGRIKTLTIDDAKIIAKQSYVASATPMTSSGGTLTYRNTDLTASLYGVGEQYFDV orfl34ng RGLKLETGRLFDENDVKEDAQWVIDQNVKDKLFADSDPLGKTILFRKRPLTVIGVMKKD I II I I I I I II II I I I I I II I I I I I I I I II II I I I I I I I I I I II I II I I II I I II I I I I I I orf134-1 RGLKLETGRLFDENDVKEDAQVWIDQNVKDKLFADSDPLGKTILFRKRPLTVIGVMKKD orfl34ng ENAFGNSDVLMLWSPYTTVMHQITGESHTNSITVKIKDNANTRVAEKGLAELLKARHGTE II I I I II I I I I I II I I II I II I II II I I I I I II I I I I I II I I: I II I I M: I II I I I I I I orf134-1 ENAFGNSDVLMLWSPYTTVMHQITGESHTNSITVKIKDNANTQVAEKGLTDLLKARHGTE orfl34ng DFFMNNSDSIRQMVESTTGTMKLLISSIALISLWGGIGVMNIMLVSVTERTKEIGIRMA I I II I I I II II I : I II II I I I I II II II I I I II I I I I I I I I II I I I I I II I I I II II I I I orf134-1 DFFMNNSDSIRQIVESTTGTMKLLISSIALISLWGGIGVMNIMLVSVTERTKEIGIRMA orfl34ng IGARRGNILQQFLIEAVLICIIGGLVGVGLSAAVSLVFNHFVTDFPMDISAASVIGAVAC I II I I I I I I I I I I I I I II I I: II I II II I I I I I II II I II I II I II I I II I I II II I I I orf134-1 IGARRGNILQQFLIEAVLICVIGGLVGVGLSAAVSLVFNHFVTDFPMDISAMSVIGAVAC orfl34ng STGIGIAFGFMPANKAAKLNPIDALAQDX II I I I I I I II I I I II II I I I I I I I I I I I I orf134-1 STGIGIAFGFMPANKAAKLNPIDALAQDX ORF134ng also shows homology to an E.coli ABC transporter: sp|P75831|YBJZ_EC0LI HYPOTHETICAL ABC TRANSPORTER ATP-BINDING PROTEIN YBJZ >gi5 (AE000189) o648; similar to YBBA_HAEIN SW: P45247 [Escherichia coli] Length = 648 Score = 297 bits (753), Expect = 6e-80 Identities = 162/389 (41%), Positives = 230/389 (58%), Gaps = 1/389 (0%) Query: 1 MSVQAVLAHKMRSLLTMLXXXXXXXXXXXXXXLGNGSQKKILEDISSMGTNTISIFPGRG 60 M+ +A+ A+KMR+LLTML +G+ +++ +L Dl S+GTNTI ++PG+ Sbjct: 260 MAWRALAANKMRTLLTMLGIIIGIASWSIVWGDAAKQMVLADIRSIGTNTIDVYPGKD 319 Query: 61 FGDRRSGKIKTLTIDDAKIIAKQSYVASATPMTSSGGTLTYRNTDLTASLYGVGEQYFDV 120 FGD + L DD I KQ +VASATP S L Y N D+ AS GV YF+V Sbjct: 320 FGDDDPQYQQALKYDDLIAIQKQPWVASATPAVSQNLRLRYNNVDVAASANGVSGDYFNV 379 Query: 121 RGLKLETGRLFDENDVKEDAQVWIDQNVKDKLFAD-SDPLGKTILFRKRPLTVIGVMKK 179 G+ G F++ + AQVW+D N + +LF +D +G+ IL P VIGV ++ Sbjct: 380 YGMTFSEGNTFNQEQLNGRAQVWLDSNTRRQLFPHKADWGEVILVGNMPARVIGVAEE 439 Query: 180 DENAFGNSDVLMLWSPYTTVMHQITGESHTNSITVKIKDNANTRVAEKGLAELLKARHGT 239 ++ FG+S VL +W PY+T+ ++ G+S NSITV++K+ ++ AE+ L LL RHG Sbjct: 440 KQSMFGSSKVLRVWLPYSTMSGRVMGQSWLNSITVRVKEGFDSAEAEQQLTRLLSLRHGK 499 Query: 240 EDFFMNNSDSIRQMVESTTGTMKXXXXXXXXXXXWGGIGVMNIMLVSVTERTKEIGIRM 299 +DFF N D + + VE TT T++ WGGIGVMNIMLVSVTERT+EIGIRM Sbjct: 500 KDFFTWNMDGVLKTVEKTTRTLQLFLTLVAVISLWGGIGVMNIMLVSVTERTREIGIRM 559 Query: 300 AIGARRGNILQQFLIEXXXXXXXXXXXXXXXXXXXXXXFNHFVTDFPMDISAASVIGAVA 359 A+GAR ++LQQFLIE F+ + + S +++ A Sbjct: 560 AVGARASDVLQQFLIEAVLVCLVGGALGITLSLLIAFTLQLFLPGWEIGFSPLALLLAFL 619 Query: 360 CSTGIGIAFGFMPANKAAKLNPIDALAQD 388 CST GI FG++PA AA+L+P+DALA++ Sbjct: 620 CSTVTGILFGWLPARNAARLDPVDALARE 648 Based on this analysis, including the presence of the leader peptide and transmembrane regions in the gonococcal protein, it is prediceted that these proteins from Nmeningitidis .and N.gonorrhoeae, and their epitopes, could be useful antigens for vaccines or diagnostics, or for raising antibodies. Example 65 The following partial DNA sequence was identified in N. meningitidis <SEQ ID 539>: 1 .. GGGACGGGAG CGATGCTGCT GCTGTTTTAC GCGGTAACGA T . CTGCCTTT 51 GGCCACTGGC GTTACCCTGA GTTACACCTC GTCGATTTTT TTGGCGGTAT 101 TTTCCTTCCT GATTTTGAAA GAACGGATTT CCGTTTACAC GCAGGCGGTG 151 CTGCTCCTTG GTTTTGCCGG CGTGGTATTG CTGCTTAATC CCTCGTTCCG 201 CAGCGGTCAG GAAACGGCGG CACTCGCCGG GCTGGCGGGC GGCGCGATGT 251 CCGGCTGGGC GTATTTGAAA GTGCGCGAAC TGTCTTTGGC GGGCGAACCC 301 GGCTGGCGCG TCGTGTTTTA CCTTTCCGTG ACAGGTGTGG CGATGTCGTC 351 GGTTTGGGCG ACGCTGACCG GCTGGCACAC CCTGTCCTTT CCATCGGCAG 401 TTTATCTGTC GTGCATCGGC GTGTCCGCGC TGATTGCCCA ACTGTCGATG 451 ACGCGCGCCT ACAAAGTCGG CGACAAATTC ACGGTTGCCT CGCTTTCCTA 501 TATGACCGTC GTTTTTTCCG CTCTGTCTGC CGCATTTTTT CTGGGCGAAG 551 AGCTTTTCTG GCAGGAAATA CTCGGTATGT GCATCATCAT C£TCAGCGGT 601 ATTTTGA This coπesponds to the .amino acid sequence <SEQ ID 540; ORF135>: l . . GΓGAWLLLFY AVTILPLATG VTLSYTSSIF LAVFSFLILK ERISVYTQAV 51 LLLGFAGWL LLNPSFRSGQ ETAALAGLAG GAMSGWAYLK VRELSLAGEP 101 GWRWFYLSV TGVAMSSVWA TLTGWHTLSF PSAVYLSCIG VSALIAQLSM 151 TRAYKVGDKF TVASLSYMTV VFSALSAAFF LGEELFWQEI LGMCIIISAV 201 F* Further work revealed the complete nucleotide sequence <SEQ ID 541>: 1 ATGGATACCG CAAAAAAAGA CATTTTAGGA TCGGGCTGGA TGCTGGTGGC 51 GGCGGCCTGC TTTACCATTA TGAACGTATT GATTAAAGAG GCATCGGCAA 101 AATTTGCCCT CGGCAGCGGC GAATTGGTCT TTTGGCGCAT GCTGTTTTCA 151 ACCGTTGCGC TCGGGGCTGC CGCCGTATTG CGTCGGGACA mCTTCCGCAC 201 GCCCCATTGG AAAAACCACT TAAACCGCAG TATGGTCGGG ACGGGGGCGA 251 TGCTGCTGCT GTTTTACGCG GTAACGCATC TGCCTTTGGC CACTGGCGTT 301 ACCCTGAGTT ACACCTCGTC GATTTTTTTG GCGGTATTTT CCTTCCTGAT 351 TTTGAAAGAA CGGATTTCCG TTTACACGCA GGCGGTGCTG CTCCTTGGTT 401 TTGCCGGCGT GGTATTGCTG CTTAATCCCT CGTTCCGCAG CGGTCAGGAA 451 ACGGCGGCAC TCGCCGGGCT GGCGGGCGGC GCGATGTCCG GCTGGGCGTA 501 TTTGAAAGTG CGCGAACTGT CTTTGGCGGG CGAACCCGGC TGGCGCGTCG 551 TGTTTTACCT TTCCGTGACA GGTGTGGCGA TGTCGTCGGT TTGGGCGACG 601 CTGACCGGCT GGCACACCCT GTCCTTTCCA TCGGCAGTTT ATCTGTCGTG 651 CATCGGCGTG TCCGCGCTGA TTGCCCAACT GTCGATGACG CGCGCCTACA 701 AAGTCGGCGA CAAATTCACG GTTGCCTCGC TTTCCTATAT GACCGTCGTT 751 TTTTCCGCTC TGTCTGCCGC ATTTTTTCTG GGCGAAGAGC TTTTCTGGCA 801 GGAAATACTC GGTATGTGCA TCATCATCCT CAGCGGTATT TTGAGCAGCA 851 TCCGCCCCAC TGCCTTCAAA CAGCGGCTGC AATCCCTGTT CCGCCAAAGA 901 TAA This coπesponds to the amino acid sequence <SEQ ID 542; ORF135-l>: 1 MDTAKKDILG SGWMLVAAAC FTIMNVLIKE ASAKFALGSG ELVFWRMLFS 51 TVALGAAAVL RRDXFRTPHW KNHLNRSMVG TGAMLLLFYA VTHLPLATGV 101 TLSYTSSIFL AVFSFLILKE RISVYTQAVL LLGFAGWLL LNPSFRSGQE 151 TAALAGLAGG AMSGWAYLKV RELSLAGEPG WRWFYLSVT GVAMSSVWAT 201 LTGWHTLSFP SAVYLSCIGV SALIAQLSMT RAYKVGDKFT VASLSYMTW 251 FSALSAAFFL GEELFWQEIL GMCIIILSGI LSSIRPTAFK QRLQSLFRQR 301 * Computer analysis of this amino acid sequence gave the following results: Homology with a predicted ORF from N.meninsitidis (sfrain A) ORF135 shows 99.0% identity over a 197aa overlap with an ORF (ORF135a) from strain A of N meningitidis: 10 20 30 orf135. pep GTGAMLLLFYAVTILPLATGVTLSYTSSIF II II I I I II I I I I I I II I I I I I I I I I I II orf135a STVALGAAAVLRRDTFRTPHWKΝHLΝRSMVGTGAMLLLFYAVTHLPLATGVTLSYTSSIF 50 60 70 80 90 100 40 50 60 70 80 90 orf135. pep LAVFSFLILKERISVYTQAVLLLGFAGWLLLΝPSFRSGQETAALAGLAGGAMSGWAYLK II I II II I I II I I II II I II I II II I I I I I I I II I I I II I II I I I I II I I II II I I I I II orf135a LAVFSFLILKERISVYTQAVLLLGFAGWLLLΝPSFRSGQETAALAGLAGGAMSGWAYLK 110 120 130 140 150 160 100 110 120 130 140 150 orf135. pep VRELSLAGEPGWRWFYLSVTGVAMSSVWATLTGWHTLSFPSAVYLSCIGVSALIAQLSM II I I I I II I II II I I I II II I I II I II I I I I I I I I II I I I I II II I I I II I II I II I II I orf135a VRELSLAGEPGWRWFYLSVTGVAMSSVWATLTGWHTLSFPSAVYLSCIGVSALIAQLSM 170 180 190 200 210 220 160 170 180 190 200 orf135. pep TRAYKVGDKFTVASLSYMTWFSALSAAFFLGEELFWQEILGMCIIISAVFX I I II I II I II I I I II II I II I I II I II I I I I : I I I I I I II I II I I I I orf135a TRAYKVGDKFTVASLSYMTVVFSALSAAFFLAEELFWQEILGMCIIILSGILSSIRPTAF 230 240 250 260 270 280 orf135a KQRLQSLFRQRX 290 300 The complete length ORF135a nucleotide sequence <SEQ ID 543> is: 1 ATGGATACCG CAAAAAAAGA CATTTTAGGA TCGGGCTGGA TGCTGGTGGC 51 GGCGGCCTGC TTTACCATTA TGAACGTATT GATTAAAGAG GCATCGGCAA 101 AATTTGCCCT CGGCAGCGGC GAATTGGTCT TTTGGCGCAT GCTGTTTTCA 151 ACCGTTGCGC TCGGGGCTGC CGCCGTATTG CGTCGGGACA CCTTCCGCAC 201 GCCCCATTGG AAAAACCACT TAAACCGCAG TATGGTCGGG ACGGGGGCGA 251 TGCTGCTGCT GTTTTACGCG GTAACGCATC TGCCTTTGGC CACCGGCGTT 301 ACCCTGAGTT ACACCTCGTC GATTTTTTTG GCGGTATTTT CCTTCCTGAT 351 TTTGAAAGAA CGGATTTCCG TTTACACGCA GGCGGTGCTG CTCCTTGGTT 401 TTGCCGGCGT GGTATTGCTG CTTAATCCCT CGTTCCGCAG CGGTCAGGAA 451 ACGGCGGCAC TCGCCGGGCT GGCGGGCGGC GCGATGTCCG GCTGGGCGTA 501 TTTGAAAGTG CGCGAACTGT CTTTGGCGGG CGAACCCGGC TGGCGCGTCG 551 TGTTTTACCT TTCCGTGACA GGTGTGGCGA TGTCATCGGT TTGGGCGACG 601 CTGACCGGCT GGCACACCCT GTCCTTTCCA TCGGCAGTTT ATCTGTCGTG 651 CATCGGCGTG TCCGCGCTGA TTGCCCAACT GTCGATGACG CGCGCCTACA 701 AAGTCGGCGA CAAATTCACG GTTGCCTCGC TTTCCTATAT GACCGTCGTT 751 TTTTCCGCTC TGTCTGCCGC ATTTTTTCTG GCCGAAGAGC TTTTCTGGCA 801 GGAAATACTC GGTATGTGCA TCATCATCCT CAGCGGTATT TTGAGCAGCA 851 TCCGCCCCAC TGCCTTCAAA CAGCGGCTGC AATCCCTGTT CCGCCAAAGA 901 TAA This encodes a protein having amino acid sequence <SEQ ID 544>: 1 MDTAKKDILG SGWMLVAAAC FTIMΝVLIKE ASAKFALGSG ELVFWRMLFS 51 TVALGAAAVL RRDTFRTPHW KΝHLΝRSMVG TGAMLLLFYA VTHLPLATGV 101 TLSYTSSIFL AVFSFLILKE RISVYTQAVL LLGFAGWLL LΝPSFRSGQE 151 TAALAGLAGG AMSGWAYLKV RELSLAGEPG WRWFYLSVT GVAMSSVWAT 201 LTGWHTLSFP SAVYLSCIGV SALIAQLSMT RAYKVGDKFT VASLSYMTW 251 FSALSAAFFL AEELFWQEIL GMCIIILSGI LSSIRPTAFK QRLQSLFRQR 301 ORF135a and ORF135-1 show 99.3% identity in 300 aa overlap: orf135a . pep MDTAKKDILGSGWMLVAAACFTIMΝVLIKEASAKFALGSGELVFWRMLFSTVALGAAAVL I II I I I I M II I I II I II I M I I M I M II I II II II I II II I II I I M M M I II I II I orf135-1 MDTAKKDILGSGWMLVAAACFTIMΝVLIKEASAKFALGSGELVFWRMLFSTVALGAAAVL orf135a. ep RRDTFRTPHWKNHLNRSMVGTGAMLLLFYAVTHLPLATGVTLSYTSSIFLAVFSFLILKE II I : II I I II I I II I I II II I I I I I I I I I II I II I I I I I II II I II I I I II I II II I I I I orf135-1 RRDXFRTPHWKNHLNRSMVGTGAMLLLFYAVTHLPLATGVTLSYTSSIFLAVFSFLILKE orf135a . pep RISVYTQAVLLLGFAGVVLLLNPSFRSGQETAALAGLAGGAMSGWAYLKVRELSLAGEPG I II II I I I I I I I I II II II I I II I II I I I II I II II I I I II II I II I I II I I II II I I I I orf135-1 RISVYTQAVLLLGFAGWLLLNPSFRSGQETAALAGLAGGAMSGWAYLKVRELSLAGEPG orf135a. pep WRWFYLSVTGVAMSΞVWATLTGWHTLSFPSAVYLSCIGVSALIAQLSMTRAYKVGDKFT I I I I I II II II II I I I I II I I I I I II I I I II I II II I I I II I I I I I I I II I I I I I II I II orf135-1 WRWFYLSVTGVAMSSVWATLTGWHTLSFPSAVYLSCIGVSALIAQLSMTRAYKVGDKFT orf135a. pep VASLSYMTWFSALSAAFFLAEELFWQEILGMCIIILSGILSSIRPTAFKQRLQSLFRQR I I I I II II II II I I I I II I I : I I II I I I II I II I II I I I I I I I I II I I I II I II II II II orf135-1 VASLSYMTWFSALSAAFFLGEELFWQEILGMCIIILSGILSSIRPTAFKQRLQSLFRQR Homology with a predicted ORF from N.sonorrhoeae ORFl 35 shows 97% identity over a 20 laa overlap with a predicted ORF (ORF135ng) from N.gonorrhoeae: orf135. pep GTGAMLLLFYAVTXLPLATGVTLSYTSSIF 30 I I II I I I I I II I I I I I : I I I I II I I I I II orfl35ng STVTLGAAAVLRRDTFRTPHWKNHLNRSMVGTGAMLLLFYAVTHLPLTTGVTLSYTSSIF 335 orf135.pep LAVFSFLILKERISVYTQAVLLLGFAGWLLLNPSFRSGQETAALAGLAGGAMSGWAYLK 90 I I I I I I II II II II I I I I I II I I II I I II II I II I I I I I I I I II I I I I I I I II I I I I II orfl35ng LAVFSFLILKERISVYTQAVLLLGFAGWLLLNPSFRSGQEPAALAGLAGGAMSGWAYLK 395 orf135.pep VRELSLAGEPGWRWFYLSVTGVAMSSVWATLTGWHTLSFPSAVYLSCIGVSALIAQLSM 150 II I II I I II II I I I II I I I : I I II I I II I II II I I II I I II II II I I I I II II I I I I I I orfl35ng VRELSLAGEPGWRWFYLSATGVAMSSVWATLTGWHTLSFPSAVYLSGIGVSALIAQLSM 455 orf135. pep TRAYKVGDKFTVASLSYMTWFSALSAAFFLGEELFWQEILGMCIIISAVF 201 II II II II II II I II II II I II I I II I I II I II I II I I II I I II II I I I : I orfl35ng TRAYKVGDKFTVASLSYMTWFSALSAAFFLGEELFWQEILGMCIIISAAF 506 An ORF135ng nucleotide sequence <SEQ ID 545> was predicted to encode a protein having amino acid sequence <SEQ ID 546>: 1 MPSEKAFRRH LRTASFQGLH LHHFHQKVGK CGIIGFGIHI FPTLLPAAQG 51 ILDIQLGLFR IDFAALAVYR RTQVDFIHTV IDGIASDQAF SEWQILRRL 101 NLGHFTDTHL IAQARRFIAD FGNIRPMRRG EAKTFCRCFR FDGIDGIHGD 151 FRQCGHINRL APGKDCRNGK RDKVFFHTRH YNQVCLEKTN CSARKIKFRH 201 Q QKKQQAAKKTTHHSSTTSS LAARFTIRPS LSQRPFMDTA KKDILGSGWM LVAAACFTVM 251 N NVVLLIIKKEEAASSAAKK FALGSGELVF WRMLFSTVTL GAAAVLRRDT FRTPHWKNHL 301 N NRRSSMMVVGGTTGGAAMM LLLFYAVTHL PLTTGVTLSY TSSIFLAVFS FLILKERISV 351 Y YTTQQAAVVLLLLLLGGFF AGWLLLNPS jTRSGQEPAAL AGLAGGAMSG WAYLKVRELS 401 LAGEPGWRW FYLSATGVAM SSVWATLTGW HTLSFPSAVY LSGIGVSALI 451 AQLSMTRAYK VGDKFTVASL SYMTWFSAL SAAFFLGEEL FWQEILGMCI 501 IISAAF* Further work revealed the following gonococcal sequence <SEQ ID 547>: 1 ATGGATACCG CAAAAAAAGA CATTTTAGGA TCGGGCTGGA TGCTGGTGGC 51 GGCGGCCTGC TTCACCGTTA TGAACGTATT GATTAAAGAG GCATCGGCAA 101 AATTTGCCCT CGGCAGCGGC GAATTGGTCT TTTGGCGCAT GCTGTTTTCA 151 ACCGTTACGC TCGGTGCTGC CGCCGTATTG CGGCGCGACA CCTTCCGCAC 201 GCCCCATTGG AAAAACCACT TAAACCGCAG TATGGTCGGG ACGGGGGCGA 251 TGCTGCTGCT GTTTTACGCG GTAACGCATC TGCCTTTGAC AACCGGCGTT 301 ACCCTGAGTT ACACCTCGTC GATTTTTttg GCGGTATTTT CCTTCCTGAT 351 TTTGAAAGAA CGGATTTCCG TTTACACGCA GGCGGTGCTG CTCCTTGGTT 401 TTGCCGGCGT GGTATTGCTG CTTAATCCCT CGTTCCGCAG CGGTCAGGAA 451 CCGGCGGCAC TCGCCGGGCT GGCGGGCGGC GCGATGTCCG GCTGGGCGTA 501 TTTGAAAGTG CGCGAACTGT CTTTGGCGGG CGAACCCGGC TGGCGCGTCG 551 TGTTTTACCT TTCCGCAACC GGCGTGGCGA TGTCGTCggt ttgggcgacg 601 Ctgaccggct ggCACAcccT GTCCTTTcca tcggcagttt ATCtgtCGGG 651 CATCGGCGTG tccgcgCtgA TTGCCCAaCT GtcgatgAcg cGCGcctaca 701 aaGTCGGCGA CAAATTCACG GTTGCCTCGC tttcctaTAt gaccgtcGTC 751 TTTTCCGCCC TGTCTGCCGC ATTTTTTCTg ggcgaagagc tttTCtggCA 801 GGAAATACTC GGTATGTGCA TCATTAtccT CAGCGGCATT TTGAGCAGCA 851 TCCGCCCCAT TGCCTTCAAA CAGCGGCTGC AAGCCCTCTT CCGCCAAAGA 901 TAA This coπesponds to the amino acid sequence <SEQ ID 548; ORF135ng-l > • 1 MDTAKKDILG SGWMLVAAAC FTVMNVLIKE ASAKFALGSG ELVFWRMLFS 51 TVTLGAAAVL RRDTFRTPHW KNHLNRSMVG TGAMLLLFYA VTHLPLTTGV 101 TLSYTSSIFL AVFSFLILKE RISVYTQAVL LLGFAGVVLL LNPSFRSGQE 151 PAALAGLAGG AMSGWAYLKV RELSLAGEPG WRWFYLSAT GVAMSSVWAT 201 LTGWHTLSFP SAVYLSGIGV SALIAQLSMT RAYKVGDKFT VASLSYMTW 251 FSALSAAFFL GEELFWQEIL GMCIIILSGI LSSIRPIAFK QRLQALFRQR 301 ORF135ng-l and ORF135-1 show 97.0% identity in 300 aa overlap: orfl35ng-l.pep MDTAKKDILGSGWMLVAAACFTVMNVLIKEASAKFALGSGELVFWRMLFSTVTLGAAAVL I II I I I I II II II I I II I I I I I: II II I I II I II I I I II II I I I II I I I I II : I I I I I I I orf135-1 MDTAKKDILGSGWMLVAAACFTIMNVLIKEASAKFALGSGELVFWRMLFSTVALGAAAVL orf135ng-l .pep RRDTFRTPHWKNHLNRSMVGTGAMLLLFYAVTHLPLTTGVTLSYTSSIFLAVFSFLILKE II I : I II I I I I II II I I I II I II I I I I II I II I I II : I II I II I I I I II II I I II II II I orf135-1 RRDXFRTPHWKNHLNRSMVGTGAMLLLFYAVTHLPLATGVTLSYTSSIFLAVFSFLILKE orf135ng-l . pep RISVYTQAVLLLGFAGWLLLNPSFRSGQEPAALAGLAGGAMSGWAYLKVRELSLAGEPG II I II I II II II II II II I II I II II II II II II II II I II II I I II I II I I I I I I II I orf135-1 RISVYTQAVLLLGFAGWLLLNPSFRSGQETAALAGLAGGAMSGWAYLKVRELSLAGEPG orf135ng-l . pep WRWFYLSATGVAMSSVWATLTGWHTLSFPSAVYLSGIGVSALIAQLSMTRAYKVGDKFT I II II I I I : I I II II II I I I I II II I I II I II I I II I II I II II I I I II I I II II I I I I orf135-1 WRVVFYLSVTGVAMSSVWATLTGWHTLSFPSAVYLSCIGVSALIAQLSMTRAYKVGDKFT orf135ng-l . pep VASLSYMTWFSALSAAFFLGEELFWQEILGMCIIILSGILSSIRPIAFKQRLQALFRQR I II I I II II II I I II I II II I II I II I I II I II I I II I I I I I I II I I I I II I I : I I I II orf135-1 VASLSYMTVVFSALSAAFFLGEELFWQEILGMCIIILSGILSSIRPTAFKQRLQSLFRQR Based on this analysis, including the presence of several putative transmembrane domains in the gonococcal protein, it is predicted that the proteins from N. meningitidis and N.gonorrhoeae, and their epitopes, could be useful antigens for vaccines or diagnostics, or for raising antibodies. Example 66 The following DNA sequence was identified in N. meningitidis <SEQ LD 549>: 1 ATGAAGCGGC GTATAGCCGT CTTCGTCCTG TTCCCGCAGA TAATCCGAGT 51 TTTGGGACAA CTGTTGCCGA AAATCGTCAA TACAGTTCCG GCACATCGGA 101 TGCTCTTCCA GATTTTCGGG ATGTTCTTTT TCTTCATACA CCAGCAATAT 151 CTGCCCGGGA TCGCCGAAAT CGATTCCCCA TGCGGCATCG TGTTCGGTGC 201 GCTCCTCTTC CGTCATCTGC CCGCGCATTG CCTGTATGGT AAAGCCGCCG 251 TAGGGGATGC CgTTGCACAC GAACATCCAG TCGCTGATGT CGTCAACCGG 301 AACGCAAACG cTTTCGCCTT GTTCGACATT GGTCAGTTCG CCsGGTTCAT 351 TGTTCAGCAC ACCGTAAATA TAAAGACCGT CAAAATAAAT ATCGTCGATC 401 CACATATGTT CGCAAATTTC GCCGTCTTCG CCGTCTTGGA AAAAAGGGAC 451 TTTGACCATG GCAAAATCCA AGGCGGAAAT AATGCGGCGG CGTTCCCAAA 501 AAAGcTCGCG CCAAAAATAT TTGAATGTTT TACGGGCGCG TTCGTCGGCA 551 CGGTTTACCG GTTCGTCTGC CTGTTCTACA TAATAAATGA CGGAATCGCC 601 CATCATATCT GCTCCTCAAC GTGTACGGTA TCTGTTTGCA CCTTACTGCG 651 GCTTTCTgcC kTCGGCATCC GATTCGGATT TGAAAAGTTC mmr yATTCG 701 GAATAG This coπesponds to the .amino acid sequence <SEQ LD 550; ORF136>: 1 MKRRIAVFVL FPQIIRVLGQ LLPKIVNTVP AHRMLFQIFG MFFFFIHQQY 51 LPGIAEIDSP CGIVFGALLF RHLPAHCLYG KAAVGDAVAH EHPVADWNR 101 NANAFALFDI GQFAXFIVQH TVNIKTVKIN IVDPHMFANF AVFAVLEKRD 151 FDHGKIQGGN NAAAFPKKLA PKIFECFTGA FVGTVYRFVC LFYIINDGIA 201 HHSAPQRVRY LFAPYCGFLP SASDSDLKSS XXSE* Further work revealed the complete nucleotide sequence <SEQ ID 551>: 1 ATGATGAAGC GGCGTATAGC CGTCTTCGTC CTGTTCCCGC AGATAATCCG 51 AGTTTTGGGA CAACTGTTGC CGAAAATCGT CAATACAGTT CCGGCACATC 101 GGATGCTCTT CCAGATTTTC GGGATGTTCT TTTTCTTCAT ACACCAGCAA 151 TATCTGCCCG GGATCGCCGA AATCGATTCC CCATGCGGCA TCGTGTTCGG 201 TGCGCTCCTC TTCCGTCATC TGCCCGCGCA TTGCCTGTAT GGTAAAGCCG 251 CCGTAGGGGA TGCCGTTGCA CACGAACATC CAGTCGCTGA TGTCGTCAAC 301 CGGAACGCAA ACGCTTTCGC CTTGTTCGAC ATTGGTCAGT TCGCCGGGTT 351 CATTGTTCAG CACACCGTAA ATATAAAGAC CGTCAAAATA AATATCGTCG 401 ATCCACATAT GTTCGCAAAT TTCGCCGTCT TCGCCGTCTT GGAAAAAAGG 451 GACTTTGACC ATGGCAAAAT CCAAGGCGGA AATAATGCGG CGGCGTTCCC 501 AAAAAAGCTC GCGCCAAAAA TATTTGAATG TTTTACGGGC GCGTTCGTCG 551 GCACGGTTTA CCGGTTCGTC TGCCTGTTCT ACATAATAAA TGACGGAATC 601 GCCCATCATT CTGCTCCTCA ACGTGTACGG TATCTGTTTG CACCTTACTG 651 CGGCTTTCTG CCTTCGGCAT CCGATTCGGA TTTGAAAAGT TCCAAATATT 701 CGGAATAG This coπesponds to the amino acid sequence <SEQ ID 552; ORF136-l>: 1 MMKRRIAVFV LFPQIIRVLG QLLPKIVNTV PAHRMLFQIF GMFFFFIHQQ 51 YLPGIAEIDS PCGIVFGALL FRHLPAHCLY GKAAVGDAVA HEHPVADWN 101 RNANAFALFD IGQFAGFIVQ HTVNIKTVKI NIVDPHMFAN FAVFAVLEKR 151 DFDHGKIQGG NNAAAFPKKL APKIFECFTG AFVGTVYRFV CLFYIINDGI 201 AHHSAPQRVR YLFAPYCGFL PSASDSDLKS SKYSE* Computer analysis of this amino acid sequence gave the following results: Homology with a predicted ORF from N.meninsitidis (strain A) ORF136 shows 71.7% identity over a 237aa overlap with an ORF (ORF136a) from strain A of N meningitidis: 10 20 30 40 50 59 orf136. pep MKRRIAVFVLFPQIIRVLGQLLPKIVΝTVPAHRMLFQIFGMFFFFIHQQYLPGIAEIDS I I I I I I II I I : I I I : I II I II I I II II I II I I I II I I I I I I I I I I I I II I I I II I I orf136a MMKRRIAVFVLLMQKIRILGQLLPKIVΝTVPAHRMLFQXFGMFFFFIHQQYLPGIAEIDS 10 20 30 40 50 60 60 70 80 90 100 110 119 orf136. pep PCGIVFGALLFRHLPAHCLYGKAAVGDAVAHEHPVADWΝRΝAΝAFALFDIGQFAXFIVQ II I II I I : I II I I : I I II I I II I I : I II II I I I I I I I I I II I I I II I I I I I I I II I I orf136a PCGIVFGTLLFRHXSTHCLYGKAAVGΝAVAHEHPVADWΝRΝAΝAFALFDIGQFAGFIVQ 70 80 90 100 110 120 120 130 140 150 160 170 179 orf136. pep HTVΝIKTVKIΝIVDPHMFAΝFAVFAVLEKRDFDHGKIQGGΝΝAAAFPKKLAPKIFECFTG M: I: I I II I II I II I I I I II I II I I II I : : I : I : I :: : : orf136a HAIΝVKTVKIΝIVDPHMFAΝFAXFAVLEKRALTMAKSKXXXMRRRSQKSSRQKYLΝVLRA 130 140 150 160 170 180 180 190 200 210 220 230 orf136. pep AFVGTVYRFVCLFYIIΝDGIAHH SAPQRVRYLFAPYCGFLPSASDSDLKSSXXSEX : M : I : : : : II II I II I II I I II II I II II I I II I II III orf136a R SPARFTGLSACSTXXMTESPIISAPQRVRYLFAPYCGFLPSASDSDLKSSKYSEX 190 200 210 220 230 The complete length ORF136a nucleotide sequence <SEQ LD 553> is: 1 ATGATGAAGC GGCGTATAGC CGTCTTCGTC CTGCTCATGC AGAAAATCCG 51 GATTTTGGGA CAACTGTTGC CGAAAATCGT CAATACAGTT CCGGCACATC 101 GGATGCTCTT CCAGATΝTTC GGGATGTTCT TTTTCTTCAT ACACCAGCAA 151 TACCTGCCCG GGATCGCCGA AATCGATTCC CCATGCGGCA TCGTGTTCGG 201 TACGCTCCTC TTCCGTCATC ΝGTCCACGCA TTGCCTGTAT GGTAAAGCCG 251 CCGTAGGGAA TGCCGTTGCA CACGAACATC CAGTCGCTGA TGTCGTCAAC 301 CGGAACGCAA ACGCTTTCGC CTTGTTCGAC ATTGGTCAGT TCGCCGGGTT 351 CATTGTTCAG CACGCCATAA ATGTAAAGAC CGTCAAAATA AATATCGTCG 401 ATCCACATAT GTTCGCAAAT TTCGCCNTCT TCGCCGTCTT GGAAAAAAGG 451 GCTTTGACCA TGGCAAAATC TAAGGNGNNA NNGATGCGGC GGCGTTCCCA 501 AAAAAGCTCG CGCCAAAAAT ATTTGAATGT TTTGCGGGCG CGTTCGCCGG 551 CACGGTTTAC CGGTTTGTCT GCCTGTTCTA CAT ATAAAT GACGGAATCG 601 CCCATCATAT CTGCTCCTCA ACGTGTACGG TATCTGTTTG CACCTTACTG 651 CGGCTTTCTG CCTTCGGCAT CCGATTCGGA TTTGAAAAGT TCCAAATATT 701 CGGAATAG This encodes a protein having amino acid sequence <SEQ LD 554>: 1 MMKRRIAVFV LLMQKIRILG QLLPKIVNTV PAHRMLFQXF GMFFFFIHQQ 51 YLPGIAEIDS PCGIVFGTLL FRHXSTHCLY GKAAVGNAVA HEHPVADWN 101 RNANAFALFD IGQFAGFIVQ HAINVKTVKI NIVDPHMFAN FAXFAVLEKR 151 ALTMAKSKXX XMRRRSQKSS RQKYLNVLRA RSPARFTGLS ACST**MTES 201 PIISAPQRVR YLFAPYCGFL PSASDSDLKS SKYSE* ORF136a and ORF136-1 show 73.1% identity in 238 aa overlap: 10 20 30 40 50 60 orf136a. pep MMKRRIAVFVLLMQKIRILGQLLPKIVNTVPAHRMLFQXFGMFFFFIHQQYLPGIAEIDS I I I II II II I I : I IM II I II I II II II I I I I I I II II II II I I I II I II II II I II orf136-1 MMKRRIAVFVLFPQIIRVLGQLLPKIVNTVPAHRMLFQIFGMFFFFIHQQYLPGIAEIDS 10 20 30 40 50 60 70 80 90 100 110 120 orf136a.pep PCGIVFGTLLFRHXSTHCLYGKAAVGNAVAHEHPVADWNRNANAFALFDIGQFAGFIVQ M I I I M : I M I I : I II I I I I I II : I II I I I I I I II I I I II I I I I I II II I II II I I I orf136-1 PCGIVFGALLFRHLPAHCLYGKAAVGDAVAHEHPVADWNRNANAFALFDIGQFAGFIVQ 70 80 90 100 110 120 130 140 150 160 170 180 orf136a . pep HAINVKTVKINIVDPHMFANFAXFAVLEKRALTMAKSKXXXMRRRSQKSSRQKYLNVLRA I :: I : I II I I I I I II I I II I I I I II I I I I : : I : I : I :: : : orf136-1 HTVNIKTVKINIVDPHMFANFAVFAVLEKRDFDHGKIQGGNNAAAFPKKLAPKIFECFTG 130 140 150 160 170 180 190 200 210 220 230 orf136a. pep R SPARFTGLSACSTXXMTESPIISAPQRVRYLFAPYCGFLPSASDSDLKSSKYSEX : I I : I : :: : II II I I I I I II I I II II I I I I II I I II I I I II I orf136-1 AFVGTVYRFVCLFYIINDGIAHH SAPQRVRYLFAPYCGFLPSASDSDLKSSKYSEX 190 200 210 220 230 Homology with a predicted ORF from N.sonorrhoeae ORF136 shows 92.3% identity over a 234aa overlap with a predicted ORF (ORF136ng) from N.gonorrhoeae: orf136.pep MKRRIAVFVLFPQIIRVLGQLLPKIVNTVPAHRMLFQIFGMFFFFIHQQYLPGIAEIDS 59 I I I II II II I : I I I M I I II II I I I I II I 1 II I II II II II II II : I I I II I II II I orfl36ng MMKRRIAVFVLLMQKIRILGQLLPKIVNTVPAHRMLFQIFGMFFFFIHRQYLPGIAEIDS 60 orf136. pep PCGIVFGALLFRHLPAHCLYGKAAVGDAVAHEHPVADWNRNANAFALFDIGQFAXFIVQ 119 I I II I I: I II I II I I I I I I I II I I I I I I I II I I I I I : I I II I I I I I II I I I I I I II orfl36ng PGGIVFGTLLFRHLSAHCLYGKAAVGDAVAHEHPVADVANRNANAFALFDIGQSAGFIVQ 120 orf136.pep HTVNIKTVKINIVDPHMFANFAVFAVLEKRDFDHGKIQGGNNAAAFPKKLAPKIFECFTG 179 I I I I II II II I I I I I II II I I I I II I I I I II I I I II II II II I I I II I I II I I: I II II I orfl36ng HTVNIKTVKINIVDPHMFANFAVFAVLEKRDFDHGKIQGGNNAAAFPKKLAPKVFECFTG 180 orf136. pep AFVGTVYRFVCLFYIINDGIAHHSAPQRVRYLFAPYCGFLPSASDSDLKSSXXSE 234 II : I II I I I I I II I II I I I I I I I : I I I I II II I I I I II I I I I I I I I II I II orfl36ng AFAGTVYRFVCLFYIINDGIAHHTAPQRVRYLFAPYRGFLPPASDSDLKSSKYSE 235 The complete length ORF136ng nucleotide sequence <SEQ LD 555> is: 1 ATGATGAAGC GGCGTATAGC CGTCTTCGTC CTGCTCATGC AGAAAATCCG 51 GATTTTGGGA CAACTGTTGC CGAAAATCGT CAATACAGTT CCGGCACATC 101 GGATGCTCTT CCAAATTTTC GGGATGTTCT TTTTCTTCAT ACACCGGCAA 151 TACCTGCCCG GGATCGCCGA AATCGATTCC CCAGGCGGTA TCGTGTTCGG 201 TACGCTCCTC TTCCGTCATC TGTCCGCGCA TTGCCTGTAC GGTAAAGCCG 251 CCGTAGGGGA TGCCGTTGCA CACGAACATC CAGTCGCTGA TGTCGCCAAC 301 CGGAACGCAA ACGCTTTCGC CTTGTTCGAC ATTGGTCAGT CCGCCGGGTT 351 CATTGTTCAG CACACCGTAA ATATAAAGAC CGTCAAAATA AATATCGTCG 401 ATCCACATAT GTTCGCAAAT TTCGCCGTCT TCGCCGTCTT GGAAAAAAGG 451 GACTTTGACC ATGGCAAAAT CCAAGGCGGA AATAATGCGG CGGCGTTCCC 501 AAAAAAGCTC GCGCCAAAAG TATTTGAATG TTTTACGGGC GCGTTCGCCG 551 GCACGGTTTA CCGGTTCGTC TGCCTGTTCT ACATAATAAA TGACGGAATC 601 GCCCATCATA CTGCTCCTCA ACGTGTACGG TATCTGTTTG CACCTTACCG 651 CGGTTTTCTA CCTCCGGCAT CCGATTCGGA TTTGAAAAGT TCCAAATATT 701 CGGAATAG This encodes a protein having amino acid sequence <SEQ ID 556>: 1 MMKRRIAVFV LLMQKIRILG QLLPKIVNTV PAHRMLFQIF GMFFFFIHRQ 51 YLPGIAEIDS PGGIVFGTLL FRHLSAHCLY GKAAVGDAVA HEHPVADVAN 101 RNANAFALFD IGQSAGFIVQ HTVNIKTVKI NIVDPHMFAN FAVFAVLEKR 151 DFDHGKIQGG NNAAAFPKKL APKVFECFTG AFAGTVYRFV CLFYIINDGI 201 AHHTAPQRVR YLFAPYRGFL PPASDSDLKS SKYSE* ORF136ng and ORF136-1 show 93.6% identity in 235 aa overlap: orfl36ng MMKRRIAVFVLLMQKIRILGQLLPKIVNTVPAHRMLFQIFGMFFFFIHRQYLPGIAEIDS I II I I II II I I : I I I : I I II II I I I II II I II II I I I I I I I I I I II : II II I II II II orf136-1 MMKRRIAVFVLFPQIIRVLGQLLPKIVNTVPAHRMLFQIFGMFFFFIHQQYLPGIAEIDS orfl36ng PGGIVFGTLLFRHLSAHCLYGKAAVGDAVAHEHPVADVANRNANAFALFDIGQSAGFIVQ I II I II: I I I I II I I II I I II I II II II II I I I I II : II I I I II I I II I I I II II I I orf136-1 PCGIVFGALLFRHLPAHCLYGKAAVGDAVAHEHPVADWNRNANAFALFDIGQFAGFIVQ orfl36ng HTVNIKTVKINIVDPHMFANFAVFAVLEKRDFDHGKIQGGNNAAAFPKKLAPKVFECFTG I II I I II I I I I I II I I I II I I I I I II II II I II I I I II I I I I I I II I I I I I I I : I I I II I orf136-1 HTVNIKTVKINIVDPHMFANFAVFAVLEKRDFDHGKIQGGNNAAAFPKKLAPKIFECFTG orfl36ng AFAGTVYRFVCLFYIINDGIAHHTAPQRVRYLFAPYRGFLPPASDSDLKSSKYSEX I I : I I II II I II I I I I I I II I II : II II I I I II II I I II I I I I I I II I II I I I I orf136-1 AFVGTVYRFVCLFYIINDGIAHHSAPQRVRYLFAPYCGFLPSASDSDLKSSKYSEX Based on the presence of the putative transmembrane domains in the gonococcal protein, it is predicted that the proteins from N. meningitidis and N.gonorrhoeae, .and their epitopes, could be useful antigens for vaccines or diagnostics, or for raising antibodies. Example 67 The following partial DNA sequence was identified in N. meningitidis <SEQ ID 557>: 1 ATGGAAAATA TGGTAACGTT TTCAAAAATC AGACCGCTTT TGGCAATCGC 51 CGCCGCCGCG TTGCTTGCCG CC . TGCGGAC GGCGGGAAAT AATGCTGTCC 101 GCAAGCCGGT GCAAACCGCC AAACCCGCCG CAGTGGTCGG TTTGGCACTC 151 GGTGGCGGCG CATCTAAAGG ATTTGCCCAT GTAGGTATTA TTAAGGTTTT 201 GAAAGAAAAC GGTATTCCTG TGAAGGTGGT TACCGGCACC TCCGCAGGTT 251 CGATTGTCGG CAACCTTTTT GCATCGGGTA TGTCGCCCGA CCGCCTCGAA 301 TTGGAAGCCG AAATTTTAGG CAAAACCGAT TTGGTCGATT TAACCTTGTC 351 CACCAATGGG TTTATCAAAG GCGCAAAGCT GCAAAATTAC ATCAACCGAA 401 AACTCCGCGG CATGCAGATT CAGCAGTTTC CCATCAAATT TGCCGCC .. This coπesponds to the amino acid sequence <SEQ ED 558; ORF137>: 1 MENMVTFSKI RPLLAIAAAA LLAAXRTAGN NAVRKPVQTA KPAAWGLAL 51 GGGASKGFAH VGIIKVLKEN GIPVKWTGT SAGSIVGNLF ASGMSPDRLE 101 LEAEILGKTD LVDLTLSTNG FIKGAKLQNY INRKLRGMQI QQFPIKFAA.. Further work revealed the complete nucleotide sequence <SEQ ID 559>: 1 ATGGAAAATA TGGTAACGTT TTCAAAAATC AGACCGCTTT TGGCAATCGC 51 CGCCGCCGCG TTGCTTGCCG CCTGCGGCAC GGCGGGAAAT AATGCTGTCC 101 GCAAGCCGGT GCAAACCGCC AAACCCGCCG CAGTGGTCGG TTTGGCACTC 151 GGTGGCGGCG CATCTAAAGG ATTTGCCCAT GTAGGTATTA TTAAGGTTTT 201 GAAAGAAAAC GGTATTCCTG TGAAGGTGGT TACCGGCACA TCGGCAGGTT 251 CGATTGTCGG CAGCCTTTTT GCATCGGGTA TGTCGCCCGA CCGCCTCGAA 301 TTGGAAGCCG AAATTTTAGG CAAAACCGAT TTGGTCGATT TAACCTTGTC 351 CACCAGTGGT TTTATCAAAG GCGAAAAGCT GCAAAATTAC ATCAACCGAA 401 AAGTCGGCGG CAGGCAGATT CAGCAGTTTC CCATCAAATT TGCCGCCGTT 451 GCTACTGATT TTGAAACCGG CAAGGCCGTC GCTTTCAATC AGGGGAATGC 501 CGGGCAGGCT GTGCGCGCTT CCGCCGCCAT TCCCAATGTG TTCCAACCCG 551 TTATCATCGG CAGGCATACA TATGTTGACG GCGGTCTGTC GCAGCCCGTG 601 CCCGTCAGTG CCGCCCGGCG GCAGGGGGCG AATTTCGTGA TTGCCGTCGA 651 TATTTCCGCC CGTCCGGGCA AAAACATCAG CCAAGGTTTC TTCTCTTATC 701 TCGATCAGAC GCTGAACGTA ATGAGCGTTT CTGCGTTGCA AAATGAGTTG 751 GGGCAGGCGG ATGTGGTTAT CAAACCGCAG GTTTTGGATT TGGGTGCAGT 801 CGGCGGATTC GATCAGAAAA AACGCGCCAT CCGGTTGGGT GAGGAGGCAG 851 CACGTGCCGC ATTGCCTGAA ATCAAACGCA AACTGGCGGC ATACCGTTAT 901 TGA This coπesponds to the amino acid sequence <SEQ ID 560; ORF137-l>: 1 MENMVTFSKI RPLLAIAAAA LLAACGTAGN NAVRKPVQTA KPAAWGLAL 51 GGGASKGFAH VGIIKVLKEN GIPVKWTGT SAGSIVGSLF ASGMSPDRLE 101 LEAEILGKTD LVDLTLSTSG FIKGEKLQNY INRKVGGRQI QQFPIKFAAV 151 ATDFETGKAV AFNQGNAGQA VRASAAIPNV FQPVIIGRHT YVDGGLSQPV 201 PVSAARRQGA NFVIAVDISA RPGKNISQGF FSYLDQTLNV MSVSALQNEL 251 GQADWIKPQ VLDLGAVGGF DQKKRAIRLG EEAARAALPE IKRKLAAYRY 301 * Computer analysis of this amino acid sequence gave the following results: Homology with a predicted ORF from N.meninsitidis (strain Al ORF137 shows 93.3% identity over a 149aa overlap with an ORF (ORF137a) from strain A of N. meningitidis: 10 20 30 40 50 60 orf137.pep MENMVTFSKIRPLLAIAAAALLAAXRTAGNNAVRKPVQTAKPAAWGLALGGGASKGFAH II I I I I II I I I II I II I I I II I I I II II I I : I II I I I II I II II I I I II I II I I I II I orfl37a MENMVTFSKIRPLLAIAAAALLAACGTAGNNAARKPVQTAKPAAWGLALGGGASKGFAH 10 20 30 40 50 60 70 80 90 100 110 120 orf13 . pep VGIIKVLKENGIPVKWTGTSAGSIVGNLFASGMSPDRLELEAEILGKTDLVDLTLSTNG I I I I II I II I II I II I I I II I I II II I : I I II I II II I II I I I I I I I I I I II II II II : I orf137a VGIIKVLKENGIPVKWTGTSAGSIVGSLFASGMSPDRLELEAEILGKTDLVDLTLSTSG 70 80 90 100 110 120 130 140 149 orf137. pep FIKGAKLQNYINRKLRGMQIQQFPIKFAA I II I II II I I I I I : I : I I I I II II I I orf137a FIKGEKLQNYINRKVGGRRIQQFPIKFAAVATDFETGKAVAFNQGNAGQAVRASAAIPNV 130 140 150 160 170 180 The complete length ORF137a nucleotide sequence <SEQ ID 561> is: 1 ATGGAAAATA TGGTAACGTT TTCAAAAATC AGACCGCTTT TGGCAATCGC 51 CGCCGCCGCG TTGCTTGCCG CCTGCGGCAC GGCGGGAAAT AATGCTGCCC 101 GCAAGCCGGT GCAAACCGCC AAACCCGCCG CAGTGGTCGG TTTGGCACTC 151 GGTGGCGGCG CATCTAAAGG ATTTGCCCAT GTAGGTATTA TTAAGGTTTT 201 GAAAGAAAAC GGTATTCCTG TGAAGGTGGT TACCGGCACA TCGGCAGGTT 251 CGATAGTCGG CAGCCTTTTT GCATCGGGTA TGTCGCCCGA CCGCCTCGAA 301 TTGGAAGCCG AAATTTTAGG TAAAACCGAT TTGGTCGATT TAACCTTGTC 351 CACCAGTGGT TTTATCAAAG GCGAAAAGCT GCAAAATTAC ATCAACCGAA 401 AAGTCGGCGG CAGGCGGATT CAGCAGTTTC CCATCAAATT TGCCGCCGTT 451 GCTACTGATT TTGAAACCGG CAAGGCCGTC GCTTTCAATC AAGGGAATGC 501 CGGGCAGGCT GTGCGCGCTT CCGCCGCCAT TCCCAATGTG TTCCAACCCG 551 TTATCATCGG CAGGCATACA TATGTTGACG GCGGTCTGTC GCAGCCCGTG 601 CCCGTCAGTG CCGCCCGGCG GCANGNNNNG NATNTCGTGA TTGCCGTCGA 651 TATTTCCGCC CGTCCGAGCA AAAACATCAG CCAAGGCTTC TTCTCTTATC 701 TCGATCAGAC GCTGAACGTA ATGAGCGTTT CCGCGTTGCA AAATGAGTTG 751 GGGCAGGCGG ATGTGGTTAT CAAACCGCAG GTTTTGGATT TGGGTGCAGT 801 CGGCGGATTC GATCAGAAAA AACGCGCCAT CCGGTTGGGT GAGGAGGCAG 851 CACGTGCCGC ATTGCCTGAA ATCAAACGCA AACTGGCGGC ATACCGTTAT 901 TGA This encodes a protein having amino acid sequence <SEQ ID 562>: 1 MENMVTFSKI RPLLAIAAAA LLAACGTAGN NAARKPVQTA KPAAWGLAL 51 GGGASKGFAH VGIIKVLKEN GIPVKWTGT SAGSIVGSLF ASGMSPDRLE 101 LEAEILGKTD LVDLTLSTSG FIKGEKLQNY INRKVGGRRI QQFPIKFAAV 151 ATDFETGKAV AFNQGNAGQA VRASAAIPNV FQPVIIGRHT YVDGGLSQPV 201 PVSAARRXXX XXVIAVDISA RPSKNISQGF FSYLDQTLNV MSVSALQNEL 251 GQADWIKPQ VLDLGAVGGF DQKKRAIRLG EEAARAALPE IKRKLAAYRY 301 ORF137a and ORF137-1 show 97.3% identity in 300 aa overlap: orf137a . pep MENMVTFSKIRPLLAIAAAALLAACGTAGNNAARKPVQTAKPAAWGLALGGGASKGFAH I I I I I I I I I I I II I I II I II I II I I I I II I II : I I II I II I I II I I I I II II II II II II orf137-1 MENMVTFSKIRPLLAIAAAALLAACGTAGNNAVRKPVQTAKPAAWGLALGGGASKGFAH orf137a . pep VGIIKVLKENGIPVKWTGTSAGSIVGSLFASGMSPDRLELEAEILGKTDLVDLTLSTSG I I I I II II I II I I I I II II I I II I I I I II I II I I I II I II II II I I I I I I II I I I II I I I orf137-1 VGIIKVLKENGIPVKWTGTSAGSIVGSLFASGMSPDRLELEAEILGKTDLVDLTLSTSG orf137a . pep FIKGEKLQNYINRKVGGRRIQQFPIKFAAVATDFETGKAVAFNQGNAGQAVRASAAIPNV I I I II I II I I I I I I I II I : I I II I I I I II I II I I I II I II I I I I I II I I I II II II II II orf137-1 FIKGEKLQNYINRKVGGRQIQQFPIKFAAVATDFETGKAVAFNQGNAGQAVRASAAIPNV orf137a . pep FQPVIIGRHTYVDGGLSQPVPVSAARRXXXXXVIAVDISARPSKNISQGFFSYLDQTLNV II I II I II I I I I II I I I II I I I I I I I I I I I I I I II I I : I I I I I I I II II II I I II orf137-1 FQPVIIGRHTYVDGGLSQPVPVSAARRQGANFVIAVDISARPGKNISQGFFSYLDQTLNV orf137 . pep MSVSALQNELGQADWIKPQVLDLGAVGGFDQKKRAIRLGEEAARAALPEIKRKLAAYRY II I I I II I II II II II I II I I I I I I II II II I I I I II I II I I I I I I I I I I II II II II I I orf137-1 MSVSALQNELGQADWIKPQVLDLGAVGGFDQKKRAIRLGEEAARAALPEIKRKLAAYRY Homology with a predicted ORF from N.sonorrhoeae ORF137 shows 89.9% identity over a 149aa overlap with a predicted ORF (ORF137ng) from N.gonorrhoeae: orf137.pep MENMVTFSKIRPLLAIAAAALLAAXRTAGNNAVRKPVQTAKPAAWGLALGGGASKGFAH 60 II I I I I I I I II : I I II I II I I II I II II I : II I I II I II I I I I: I I I II I II I I I II orfl37ng MENMVTFSKIRSFLAIAAAALLAACGTAGNNAARKPVQTAKPAAWALALGGGASKGFAH 60 orf137.pep VGIIKVLKENGIPVKWTGTSAGSIVGNLFASGMSPDRLELEAEILGKTDLVDLTLSTNG 120 : I I: I I I II I II II II I I I II I I I I I I : I : II I I I I I I I I I I I II I I I II I II I I I I I : I orfl37ng IGIVKVLKENGIPVKWTGTSAGSIVGSLLASGMSPDRLELEAEILGKTDLVDLTLSTSG 120 orf137. pep FIKGAKLQNYINRKLRGMQIQQFPIKFAA 149 I I II II I II I I I I : I II I II I I I I II orfl37ng FIKGEKLQNYINRKVGGRQIQQFPIKFAAVATDFETGKAVAFNQGNAGQAVRASAAIPNV 180 The complete length ORF137ng nucleotide sequence <SEQ ID 563> is: 1 ATGGAAAATA TGGTAACGTT TTCAAAAATC AGATCATTTT TGGCAATCGC 51 CGCCGCCGCG TTGCTTGCCG CCTGCGGTAC GGCGGGAAAC AATGCCGCCC 101 GCAAGCCGGT GCAAACCGCC AAACCCGCCG CAGTGGTCGC TTTGGCACTC 151 GGTGGCGGCG CATCTAAAGG ATTTGCCCAT ATAGGAATTG TTAAGGTTTT 201 GAAAGAAAAC GGTATTCCTG TGAAGGTGGT TACCGGCACA TCGGCAGGTT 251 CGA AGTCGG CAGCCTTTTG GCATCGGGTA TGTCGCCCGA CCGCCTCGAA 301 TTGGAAGCCG AGATTTTAGG TAAAACCGAT TTAGTCGATT TAACCTTGTC 351 CACCAGTGGT TTTATCAAAG GCGAAAAGCT GCAAAATTAC ATCAACCGAA 401 AAGTCGGCGG CAGGCAGATT CAGCAGTTTC CCATCAAATT TGCCGCCGTT 451 GCCACTGATT TTGAAACCGG CAAGGCCGTC GCTTTCAATC AAGGGAATGC 501 CGGGCAGGCG GTTCGTGCTT CCGCCGCCAT TCCCAATGTG TTCCAGCCAG 551 TCATCATCGG CAGGCACAAA TATGTTGACG GCGGTCTGTC GCAGCCCGTG 601 CCCGTCAGTG CCGCTCGGCG GCAGGGGGCG AATTTCGTGA TTGCCGTCGA 651 TATTTCCGCA CGTCCGAGCA AAAATGTCGG TCAAGGTTTC TTCTCTTATC 701 TCGATCAGAC GCTGAACGTG ATGAGCGTTT CCGTGTTGCA AAACGAGTTG 751 gggcAGGCGG ATGTGGTTAT CAAACCGCag gtTTTGGATT TGGGTGCAGT 801 CGGCGGATTC GATCAGAAAA AGCGCGCCAT CCGGTTGGGC GAGGAGGCAG 851 CACGTGCCGC ATTGCCTGAA ATCAAACGCA AACTGGCGGC ATACCGTTAT 901 TGA This encodes a protein having amino acid sequence <SEQ ID 564>: 1 MENMVTFSKI RSFLAIAAAA LLAACGTAGN NAARKPVQTA KPAAWALAL 51 GGGASKGFAH IGIVKVLKEN GIPVKWTGT SAGSIVGSLL ASGMSPDRLE 101 LEAEILGKTD LVDLTLSTSG FIKGEKLQNY INRKVGGRQI QQFPIKFAAV 151 ATDFETGKAV AFNQGNAGQA VRASAAIPNV FQPVIIGRHK YVDGGLSQPV 201 PVSAARRQGA NFVIAVDISA RPSKNVGQGF FSYLDQTLNV MSVSVLQNEL 251 GQADWIKPQ VLDLGAVGGF DQKKRAIRLG EEAARAALPE IKRKLAAYRY 301 * ORF137ng and ORF137-1 show 96.0% identity in 300 aa overlap: orfl37ng MENMVTFSKIRSFLAIAAAALLAACGTAGNNAARKPVQTAKPAAWALALGGGASKGFAH I I I II I II I II : I II I II I II II I I I I I I II : II II I II I I I II I : I I I II I I I I II I I orf137-1 MENMVTFSKIRPLLAIAAAALLAACGTAGNNAVRKPVQTAKPAAWGLALGGGASKGFAH orfl37ng IGIVKVLKENGIPVKVVTGTSAGSIVGSLLASGMSPDRLELEAEILGKTDLVDLTLSTSG : I I: I I II I II I I I II I II I I I II I I I I I : I II I I II I I I I I I I I I II I I I II I I I I II I orf137-1 VGIIKVLKENGIPVKWTGTSAGSIVGSLFASGMSPDRLELEAEILGKTDLVDLTLSTSG orfl37ng FIKGEKLQNYINRKVGGRQIQQFPIKFAAVATDFETGKAVAFNQGNAGQAVRASAAIPNV II I II I II I I I I II I II I II I I I II I II I II I II II I II I I I I I II I I I II I I II II I II orf137-1 FIKGEKLQNYINRKVGGRQIQQFPIKFAAVATDFETGKAVAFNQGNAGQAVRASAAIPNV orfl37ng FQPVIIGRHKYVDGGLSQPVPVSAARRQGANFVIAVDISARPSKNVGQGFFSYLDQTLNV II I II I I II II I II I I I I II II I I II II I II I II I I I II I I : IM: I I I I II I I I I II I orf137-1 FQPVIIGRHTYVDGGLSQPVPVSAARRQGANFVIAVDISARPGKNISQGFFSYLDQTLNV orfl37ng MSVSVLQNELGQADWIKPQVLDLGAVGGFDQKKRAIRLGEEAARAALPEIKRKLAAYRY I II I M I I II I II II I II I I II II I I II II I II I I II II I II II I I I I I II I II II I II I orf137 MSVSALQNELGQADWIKPQVLDLGAVGGFDQKKRAIRLGEEAARAALPEIKRKLAAYRY Based on the presence of a predicted prokaryotic membrane lipoprotein lipid attachment site (underlined) in the gonococcal protein, it is predicted that the proteins from N. meningitidis .and N.gonorrhoeae, .and their epitopes, could be useful antigens for vaccines or diagnostics, or for raising antibodies. Example 68 The following partial DNA sequence was identified in N. meningitidis <SEQ ID 565>: 1 ATGTTTCGTT TACAATTCAG GCTGTTTCCC CCTTTGCGAA CCGCCATGCA 51 CATCCTGTTG ACCGCCCTGC TCAAATGCCT CTCCCTGcTG CCGCTTTCCT 101 GTCTGCACAC GCTGGGAAAC CGGCTCGGAC ATCTGGCGTT TTACCTTTTA 151 AAGGAAGACC GCGCGCGCAT CGTCGCCmAT ATGCGGCAGG CGGGTTTGAA 201 CCCCGACCCC AAAACGGTCA AAGCCGTTTT TGCGGAAACG GCAAAAGGCG 251 GTTTGGAACT TGCCCCCGCG TTTTTCAGAA AACCGGAAGA CATAGAAACA 301 ATGTTCAAAG CGGTACACGG CTGGGAACAT GTGCAGCAGG CTTTGGACAA 351 ACACGAAGGG CTGCTATTC .. This coπesponds to the amino acid sequence <SEQ ID 566; ORF138>: 1 MFRLQFRLFP PLRTAMHILL TALLKCLSLL PLSCLHTLGN RLGHLAFYLL 51 KEDRARIVAX MRQAGLNPDP KTVKAVFAET AKGGLELAPA FFRKPEDIET 101 MFKAVHGWEH VQQALDKHEG LLF Further work revealed the complete nucleotide sequence <SEQ ID 567>: 1 ATGTTTCGTT TACAATTCAG GCTGTTTCCC CCTTTGCGAA CCGCCATGCA 51 CATCCTGTTG ACCGCCCTGC TCAAATGCCT CTCCCTGCTG CCGCTTTCCT 101 GTCTGCACAC GCTGGGAAAC CGGCTCGGAC ATCTGGCGTT TTACCTTTTA 151 AAGGAAGACC GCGCGCGCAT CGTCGCCAAT ATGCGGCAGG CGGGTTTGAA 201 CCCCGACCCC AAAACGGTCA AAGCCGTTTT TGCGGAAACG GCAAAAGGCG 251 GTTTGGAACT TGCCCCCGCG TTTTTCAGAA AACCGGAAGA CATAGAAACA 301 ATGTTCAAAG CGGTACACGG CTGGGAACAT GTGCAGCAGG CTTTGGACAA 351 ACACGAAGGG CTGCTATTCA TCACGCCGCA CATCGGCAGC TACGATTTGG 401 GCGGACGCTA CATCAGCCAG CAGCTTCCGT TCCCGCTGAC CGCCATGTAC 451 AAACCGCCGA AAATCAAAGC GATAGACAAA ATCATGCAGG CGGGCAGGGT 501 TCGCGGCAAA GGAAAAACCG CGCCTACCAG CATACAAGGG GTCAAACAAA 551 TCATCAAAGC CCTGCGTTCG GGCGAAGCAA CCATCGTCCT GCCCGACCAC 601 GTCCCCTCCC CTCAAGAAGG CGGGGAAGGC GTATGGGTGG ATTTCTTCGG 651 CAAACCTGCC TATACCATGA CGCTGGCGGC AAAATTGGCA CACGTCAAAG 701 GCGTGAAAAC CCTGTTTTTC TGCTGCGAAC GCCTGCCTGG CGGACAAGGT 751 TTCGATTTGC ACATCCGCCC CGTCCAAGGG GAATTGAACG GCGACAAAGC 801 CCATGATGCC GCCGTGTTCA ACCGCAATGC CGAATATTGG ATACGCCGTT 851 TTCCGACGCA GTATCTGTTT ATGTACAACC GCTACAAAAT GCCGTAA This coπesponds to the amino acid sequence <SEQ ID 568; ORF138-l>: 1 MFRLQFRLFP PLRTAMHILL TALLKCLSLL PLSCLHTLGN RLGHLAFYLL 51 KEDRARIVAN MRQAGLNPDP KTVKAVFAET AKGGLELAPA FFRKPEDIET 101 MFKAVHGWEH VQQALDKHEG LLFITPHIGS YDLGGRYISQ QLPFPLTAMY 151 KPPKIKAIDK IMQAGRVRGK GKTAPTSIQG VKQIIKALRS GEATIVLPDH 201 VPSPQEGGEG VWVDFFGKPA YTMTLAAKLA HVKGVKTLFF CCERLPGGQG 251 FDLHIRPVQG ELNGDKAHDA AVFNRNAEYW IRRFPTQYLF MYNRYKMP* Computer analysis of this amino acid sequence gave the following results: Homology with a predicted ORF from N.meninsitidis (sfrain Al ORF138 shows 99.2% identity over a 123aa overlap with an ORF (ORF138a) from strain A of N meningitidis: 10 20 30 40 50 60 orf138. pep MFRLQFRLFPPLRTAMHILLTALLKCLSLLPLSCLHTLGΝRLGHLAFYLLKEDRARIVAX II I I I I I II II I I I I II II I I II I I II I II II I II I II I I I I I II I I I I I II II I I I II orf138a MFRLQFRLFPPLRTAMHILLTALLKCLSLLPLSCLHTLGΝRLGHLAFYLLKEDRARIVAΝ 10 20 30 40 50 60 70 80 90 100 110 120 orf138. pep MRQAGLΝPDPKTVKAVFAETAKGGLELAPAFFRKPEDIETMFKAVHGWEHVQQALDKHEG II II I I I II II I I I I II I I I I II I I I I I II I I I I II I II I I I I II II I I I II I II II II I orf138a MRQAGLΝPDPKTVKAVFAETAKGGLELAPAFFRKPEDIETMFKAVHGWEHVQQALDKHEG 70 80 90 100 110 120 orf138. pep LLF I I I orf138a LLFITPHIGSYDLGGRYISQQLPFPLTAMYKPPKIKAIDKIMQAGRVRGKGKTAPTSIQG 130 140 150 160 170 180 The complete length ORF138a nucleotide sequence <SEQ ED 569> is: 1 ATGTTTCGTT TACAATTCAG GCTGTTTCCC CCTTTGCGAA CCGCCATGCA 51 CATCCTGTTG ACCGCCCTGC TCAAATGCCT CTCCCTGCTG CCGCTTTCCT 101 GTCTGCACAC GCTGGGAAAC CGGCTCGGAC ATCTGGCGTT TTACCTTTTA 151 AAGGAAGACC GCGCGCGCAT CGTCGCCAAT ATGCGTCAGG CAGGCATGAA 201 TCCCGACCCC AAAACGGTCA AAGCCGTTTT TGCGGAAACG GCAAAAGGCG 251 GTTTGGAACT TGCCCCCGCG TTTTTCAGAA AACCGGAAGA CATAGAAACA 301 ATGTTCAAAG CGGTACACGG CTGGGAACAT GTGCAGCAGG CTTTGGACAA 351 ACACGAAGGG CTGCTATTCA TCACGCCGCA CATCGGCAGC TACGATTTGG 401 GCGGACGCTA CATCAGCCAG CAGCTTCCGT TCCCGCTGAC CGCCATGTAC 451 AAACCGCCGA AAATCAAAGC GATAGACAAA ATCATGCAGG CGGGCAGGGT 501 TCGCGGCAAA GGAAAAACCG CGCCTACCAG CATACAAGGG GTCAAACAAA 551 TCATCAAAGC CCTGCGTTCG GGCGAAGCAA CCATCGTCCT GCCCGACCAC 601 GTCCCCTCCC CTCAAGAAGG CGGGGAAGGC GTATGGGTGG ATTTCTTCGG 651 CAAACCTGCC TATACCATGA CGCTGGCGGC AAAATTGGCA CACGTCAAAG 701 GCGTGAAAAC CCTGTTTTTC TGCTGCGAAC GCCTGCCTGG CGGACAAGGT 751 TTCGATTTGC ACATCCGCCC CGTCCAAGGG GAATTGAACG GCGACAAAGC 801 CCATGATGCC GCCGTGTTCA ACCGCAATGC CGAATATTGG ATACGCCGTT 851 TTCCGACGCA GTATCTGTTT ATGTACAACC GCTACAAAAT GCCGTAA This encodes a protein having amino acid sequence <SEQ ID 570>: 1 MFRLQFRLFP PLRTAMHILL TALLKCLSLL PLSCLHTLGN RLGHLAFYLL 51 KEDRARIVAN MRQAGLNPDP KTVKAVFAET AKGGLELAPA FFRKPEDIET 101 MFKAVHGWEH VQQALDKHEG LLFITPHIGS YDLGGRYISQ QLPFPLTAMY 151 KPPKIKAIDK IMQAGRVRGK GKTAPTSIQG VKQIIKALRS GEATIVLPDH 201 VPSPQEGGEG VWVDFFGKPA YTMTLAAKLA HVKGVKTLFF CCERLPGGQG 251 FDLHIRPVQG ELNGDKAHDA AVFNRNAEYW IRRFPTQYLF MYNRYKMP* ORF138a and ORF138-1 show 99.7% identity over a 298aa overlap: orf138a . pep MFRLQFRLFPPLRTAMHILLTALLKCLSLLPLSCLHTLGNRLGHLAFYLLKEDRARIVAN I II I I I I I I II II II I II II I II I I I II II II I II II I I I I II II II II I II I I I I I I II orf138-1 MFRLQFRLFPPLRTAMHILLTALLKCLSLLPLSCLHTLGNRLGHLAFYLLKEDRARIVAN orf138a . pep MRQAGMNPDPKTVKAVFAETAKGGLELAPAFFRKPEDIETMFKAVHGWEHVQQALDKHEG I I II I : I I I II II I II I I I II I II I I II I II II I II II I I II II II I I I II II II II I I I orf138-1 MRQAGLNPDPKTVKAVFAETAKGGLELAPAFFRKPEDIETMFKAVHGWEHVQQALDKHEG orf138a . pep LLFITPHIGSYDLGGRYISQQLPFPLTAMYKPPKIKAIDKIMQAGRVRGKGKTAPTSIQG I II I I I II I I I II II I II II I I I I I I II II I II I II II I I II II II I I I I I II II II I I I orf138-1 LLFITPHIGSYDLGGRYISQQLPFPLTAMYKPPKIKAIDKIMQAGRVRGKGKTAPTSIQG orf138a . pep VKQIIKALRSGEATIVLPDHVPSPQEGGEGVWVDFFGKPAYTMTLAAKLAHVKGVKTLFF I I I I I I II II I II II I II II I I I I I I II I II II I I I I I I I I I II II I I I II I II II I I I I orf138-1 VKQIIKALRSGEATIVLPDHVPSPQEGGEGVWVDFFGKPAYTMTLAAKLAHVKGVKTLFF or 138a . pep CCERLPGGQGFDLHIRPVQGELNGDKAHDAAVFNRNAEYWIRRFPTQYLFMYNRYKMP I II I II I I I II II I II I I I I I I II I I I II II I I I I I I I I II I I I I I I I I I II I I I II I orf138-1 CCERLPGGQGFDLHIRPVQGELNGDKAHDAAVFNRNAEYWIRRFPTQYLFMYNRYKMP Homology with a predicted ORF from Nsonorrhoeae ORF138 shows 94.3% identity over a 123aa overlap with a predicted ORF (ORF138ng) from N.gonorrhoeae: orf138.pep MFRLQFRLFPPLRTAMHILLTALLKCLSLLPLΞCLHTLGNRLGHLAFYLLKEDRARIVAX 60 I I II I I I I I I II I II I II II I I II I I II I I I II I II I I I I II II II I I I II I I II II I orf138ng MFRLQFRLFPPLRTAMHILLTALLKCLSLLSLSCLHTLGNRLGHLAFYLLKEDRARIVAN 60 orf138.pep MRQAGLNPDPKTVKAVFAETAKGGLELAPAFFRKPEDIETMFKAVHGWEHVQQALDKHEG 120 I II I I I I I I : II I II I I I I II II I II I I I I : I II II I I I I I I I I II I I I I II I I I II orf138ng MRQAGLNPDTQTVKAVFAETAKCGLELAPAFFKKPEDIETMFKAVHGWEHVQQALDKGEG 120 orfl38.pep LLF 123 I I I orfl38ng LLFITPHIGSYDLGGRYISQQLPFHLTAMYKPPKIKAIDKIMQAGRVRGKGKTAPTGIQG 180 The complete length ORF138ng nucleotide sequence <SEQ LD 571> is: 1 ATGTTTCGTT TACAATTCAG GCTGTTTCCC CCTTTGCGAA CCGCCATGCA 51 CATCCTGTTG ACCGCCCTGC TCAAATGCCT CTCCCTGCTG TCGCTTTCCT 101 GTCTGCACAC GCTGGGAAAC CGGCTCGGAC ATCTGGCGTT TTACCTTTTA 151 AAGGAAGACC GCGCGCGCAT CGTCGCCAAT ATGCGGCAGG CGGGTTTGAA 201 CCCCGACACG CAGACGGTCA AAGCCGTTTT TGCGGAAACG GCAAAATGCG 251 GTTTGGAACT TGCCCCCGCG TTTTTCAAAA AACCGGAAGA CATCGAAACA 301 ATGTTCAAAG CGGTACACGG CTGGGAACAC GTGCAGCAGG CTTTGGACAA 351 GGGCGAAGGG CTGCTGTTCA TCACGCCGCA CATCGGCAGC TACGATTTGG 401 GCGGACGCTA CATCAGCCAG CAGCTTCCGT TCCACCTGAC CGCCATGTAC 451 AAGCCGCCGA AAATCAAAGC GATAGACAAA ATCATGCAGG CGGGCAGGGT 501 GCGCGGCAAA GGCAAAACcg cgcccaccgg catACAAGGG GTCAAACAAA 551 tcatcaAGGC CCTGCGCGCG GGCGAGGCAA CCAtcATCCT GCCCGACCAC 601 GTCCCTTCTC CGCAGGAagg cggCGGCGTG TGGGCGGATT TTTTCGGCAA 651 ACCTGCATAc acCATGACAC TGGCGGCAAA ATTGGCACAC GTCAAAGGCG 701 TGAAAACCCT GTTTTTCTGC TGCGAACGCC TGCCCGACGG ACAAGGCTTC 751 GTGTTGCACA TCCGCCCCGT CCAAGGGGAA TTGAACGGCA ACAAAGCCCA 801 CGATGCCGCC GTGTTCAACC GCAATACCGA ATATTGGATA CGCCGTTTTC 851 CGACGCAGTA TCTGTTTATG TACAACCGCT ATAAAACGCC GTAA This encodes a protein having amino acid sequence <SEQ ID 572>: 1 MFRLQFRLFP PLRTAMHILL TALLKCLSLL SLSCLHTLGN RLGHLAFYLL 51 KEDRARIVAN MRQAGLNPDT QTVKAVFAET AKCGLELAPA FFKKPEDIET 101 MFKAVHGWEH VQQALDKGEG LLFITPHIGS YDLGGRYISQ QLPFHLTAMY 151 KPPKIKAIDK IMQAGRVRGK GKTAPTGIQG VKQIIKALRA GEATIILPDH 201 VPSPQEGGGV WADFFGKPAY TMTLAAKLAH VKGVKTLFFC CERLPDGQGF 251 VLHIRPVQGE LNGNKAHDAA VFNRNTEYWI RRFPTQYLFM YNRYKTP* ORF138ng and ORF138-1 show 94.3% identity over 299aa overlap: orf138-1.pep MFRLQFRLFPPLRTAMHILLTALLKCLSLLPLSCLHTLGNRLGHLAFYLLKEDRARIVAN II I II II I I I I II I I I II I II I II I I II II II I I I I II II II I I I II I I I II II I I I II orfl38ng MFRLQFRLFPPLRTAMHILLTALLKCLSLLSLSCLHTLGNRLGHLAFYLLKEDRARIVAN orf138-1. pep MRQAGLNPDPKTVKAVFAETAKGGLELAPAFFRKPEDIETMFKAVHGWEHVQQALDKHEG II I II II I I : II I I I II I II I II I II I I I I : I II I I I I I I I I I II I I I II I I II I II orfl38ng MRQAGLNPDTQTVKAVFAETAKCGLELAPAFFKKPEDIETMFKAVHGWEHVQQALDKGEG orf138-1. pep LLFITPHIGSYDLGGRYISQQLPFPLTAMYKPPKIKAIDKIMQAGRVRGKGKTAPTSIQG II II I II I II I II I I I II I II I I I I II I II I II II II II II II I II I I II I I I II : II I orfl38ng LLFITPHIGSYDLGGRYISQQLPFHLTAMYKPPKIKAIDKIMQAGRVRGKGKTAPTGIQG orf138-1. pep VKQIIKALRSGEATIVLPDHVPSPQEGGEGVWVDFFGKPAYTMTLAAKLAHVKGVKTLFF I I II II I II : I II I I: I I I II I I I I I II II I : I II I I II II II I I I I I II I I I II II I I orfl38ng VKQIIKALRAGEATIILPDHVPSPQEGG-GVWADFFGKPAYTMTLAAKLAHVKGVKTLFF orf138-1. pep CCERLPGGQGFDLHIRPVQGELNGDKAHDAAVFNRNAEYWIRRFPTQYLFMYNRYKMP I II II I I I I I I I I II I II I I II : II I II I II II I : II II I I I I I I I II II I II I I orfl38ng CCERLPDGQGFVLHIRPVQGELNGNKAHDAAVFNRNTEYWIRRFPTQYLFMYNRYKTP In addition, ORF138ng is homologous to htrB protein from Pseudomonas fluorescens: gnl|PID|e334283 (Y14568) htrB [Pseudomonas fluorescens] Length = 253 Score = 80.8 bits (196), Expect = 9e-15 Identities = 49/151 (32%), Positives = 79/151 (51%), Gaps = 6/151 (3%) Query: 101 MFKAVHGWEHVQQALDKGEGLLFITPHIGSYD-LGGRYISQQLPFHLTAMYKPPKIKAID 159 + + V G E +++AL G+G++ IT H+G+++ L Y SQ P Y+PPK+KA+D Sbjct: 94 LVREVEGLEVLKEALASGKGWGITSHLGNWEVLNHFYCSQCKPI IFYRPPKLKAVD 150 Query: 160 KIMQAGRVRGKGKTAPTGIQGVKQIIKALRAGEATIILPDHVPSPQEGGGVWADFFGKPA 219 ++++ RV+ K A + +G+ +IK +R G I D P P E G++ FF A Sbjct: 151 ELLRKQRVQLGNKVAASTKEGILSVIKEVRKGGQVGIPAD—PEPAESAGIFVPFFATQA 208 Qu-^ry: 220 YTMTLAAKLAHVKGVKTLFFCCERLPDGQGF 250 T + +F RLPDG G+ Sbjct: 209 LTSKFVPNMLAGGKAVGVFLHALRLPDGSGY 239 Based on this analysis, including the presence of a putative transmembrane domain in the gonococcal protein, it was predicted that the proteins from N. meningitidis and N.gonorrhoeae, and their epi + opes, could be useful antigens for vaccines or diagnostics, or for raising antibodies. ORF138-1 (57kDa) was cloned in the pGex vectors and expressed in E.coli, as described above. The products of protein expression and purification were analyzed by SDS-PAGE. Figure 14A shows the results of affinity purification of the GST-fusion protein. Purified GST- fusion protein was used to immunise mice, whose sera were used for ELISA (positive result) and FACS analysis (Figure 14B). These experiments confirm that ORF138-1 is a surface-exposed protein, and that it is a useful immunogen. Example 69 The following partial DNA sequence was identified in N. meningitidis <SEQ ED 573>: 1 ..GCGTGGTCGG CCGGCGAATC GTGGCGTGTG TTAATGGAAA GTGAAACGTG 51 GCATGCGGTG TGGAATACTT TGCGCTTCTC GGCGGCGGCG GTGTATGCGG 101 CAGCGGTTTT GGGTGTGGTG TATGCGGCGC CGGCGCGGCG GTCGGCGTGG 151 ATGCGCGGGC TGATGTTTTA GCCGTTTATG GTGTCGCCGG TTTGTGTTTC 201 GGCGGGCGTG CTGCTGCTTT ATCCGCAGTG GACGGCTTCG TTGCCGTTGC 251 TGCTGGCGAT GTATGCGCTG CTGGCGTATC CGTTTGTGGC AAAAGATGTT 301 TTATCAGCCT GGGATGCACT GCCGCCGGAT TACGGCAGGG CGGCGGCGGG 351 TTTGGGTGCA AACGGCTTTC AGACGGCATG CCGCATCACG TTCCCCCTCT 401 TGAAACCGGC GTTGCGGCGC GGTCTGACTT TGGCGGCGGC AACCTGCGTG 451 GGCGAATTTG CGGCGACATT GTTTCTGTCG CGTCCGGAAT GGCAGACGCT 501 GACGACTTTG ATTTATGCCT ATTTGGGACG CGCGGGTGAG GATAATTACG 551 CGCGGGCGAT GGTGCTG.. This coπesponds to the amino acid sequence <SEQ ID 574; ORF139>: 1 .. AWSAGESWRV LMESETWHAV WNTLRFSAAA VYAAAVLGW YAAPARRSAW 51 MRGLMFXPFM VSPVCVSAGV LLLYPQWTAS LPLLLAMYAL LAYPFVAKDV 101 LSAWDALPPD YGRAAAGLGA NGFQTACRIT FPLLKPALRR GLTLAAATCV 151 GEFAATLFLS RPEWQTLTTL IYAYLGRAGE DNYARAMVX.. Further work revealed the complete nucleotide sequence <SEQ ID 575>: 1 ATGGATGGAC GGCGTTGGGT GGTATGGGGT GCTTTTGCCC TGCTGCCTTC 51 GGCTTTTTTG GCGGTAATGG TCGTTGCGCC TTTGTGGGCG GTGGCGGCGT 101 ATGACGGTTT GGCGTGGCGC GCGGTGCTGT CGGATGCCTA TATGCTCAAA 151 CGTTTGGCGT GGACGGTATT TCAGGCAGCG GCAACCTGTG TGCTGGTGCT 201 GCCTTTGGGC GTGCCTGTCG CGTGGGTGCT GGCGCGGCTG GCGTTTCCGG 251 GGCGGGCTTT GGTGCTGCGC CTGCTGATGC TGCCTTTTGT GATGCCCACG 301 TTGGTGGCGG GCGTGGGCGT GCTGGCCCTG TTCGGGGCGG ACGGGCTGTT 351 GTGGCGCGGC AGGCAGGATA CGCCGTATCT GTTGTTGTAC GGCAATGTGT 401 TTTTCAACCT TCCTGTGTTG GTCAGGGCGG CGTATCAGGG GTTTGTGCAA 451 GTGCCTGCGG CACGGCTTCA GACGGCACGG ACGTTGGGCG CGGGGGCGTG 501 GCGGCGGTTT TGGGACATTG AAATGCCCGT TTTGCGCCCG TGGCTTGCCG 551 GCGGCGTGTG CCTTGTCTTT CTGTATTGTT TTTCCGGGTT CGGGCTGGCG 601 CTGCTGCTGG GCGGCAGCCG TTATGCCACG GTCGAAGTGG AAATTTACCA 651 GTTGGTCATG TTCGAACTCG ATATGGCGGT TGCTTCGGTG CTGGTGTGGC 701 TGGTGTTGGG GGTAACGGCG GCGGCAGGGT TGCTGTATGC GTGGTTCGGC 751 AGGCGCGCGG TTTCGGATAA GGCGGTTTCC CCTGTGATGC CGTCGCCGCC 801 GCAGTCGGTC GGGGAATATG TGCTGCTGGC GTTTGCGGCG GCGGTGTTGT 851 CTGTGTGCTG CCTGTTTCCT TTGTTGGCAA TTGTTGTGAA AGCGTGGTCG 901 GCCGGCGAAT CGTGGCGTGT GTTAATGGAA AGTGAAACGT GGCAGGCGGT 951 GTGGAATACT TTGCGCTTCT CGGCGGCGGC GGTGTATGCG GCGGCGGTTT 1001 TGGGTGTGGT GTATGCGGCG GCGGCGCGGC GGTCGGCGTG GATGCGCGGG 1051 CTGATGTTTT TGCCGTTTAT GGTGTCGCCG GTTTGTGTTT CGGCGGGCGT 1101 GCTGCTGCTT TATCCGCAGT GGACGGCTTC GTTGCCGTTG CTGCTGGCGA 1151 TGTATGCGCT GCTGGCGTAT CCGTTTGTGG CAAAAGATGT TTTATCAGCC 1201 TGGGATGCAC TGCCGCCGGA TTACGGCAGG GCGGCGGCGG GTTTGGGTGC 1251 AAACGGCTTT CAGACGGCAT GCCGCATCAC GTTCCCCCTC TTGAAACCGG 1301 CGTTGCGGCG CGGTCTGACT TTGGCGGCGG CAACCTGCGT GGGCGAATTT 1351 GCGGCGACAT TGTTTCTGTC GCGTCCGGAA TGGCAGACGC TGACGACTTT 1401 GATTTATGCC TATTTGGGAC GCGCGGGTGA GGATAATTAC GCGCGGGCGA 1451 TGGTGCTGAC ATTGCTGTTG GCGGCGTTCG CGCTGGGTAT TTTCCTGCTG 1501 TTGGACGGCG GCGAAGGCGG AAAACAGACG GAAACGTTAT AA This coπesponds to the .amino acid sequence <SEQ ID 576; ORF139-l>: 1 MDGRRWWWG AFALLPSAFL AVMWAPLWA VAAYDGLAWR AVLSDAYMLK 51 RLAWTVFQAA ATCVLVLPLG VPVAWVLARL AFPGRALVLR LLMLPFVMPT 101 LVAGVGVLAL FGADGLLWRG RQDTPYLLLY GNVFFNLPVL VRAAYQGFVQ 151 VPAARLQTAR TLGAGAWRRF WDIEMPVLRP WLAGGVCLVF LYCFSGFGLA 201 LLLGGSRYAT VEVEIYQLVM FELDMAVASV LVWLVLGVTA AAGLLYAWFG 251 RRAVSDKAVS PVMPSPPQSV GEYVLLAFAA AVLSVCCLFP LLAIWKΛffS 301 AGESWRVLME SETWQAVWNT LRFSAAAVYA AAVLGWYAA AARRSAWMRG 351 LMFLPFMVSP VCVSAGVLLL YPQWTASLPL LLAMYALLAY PFVAKDVLSA 401 WDALPPDYGR AAAGLGANGF QTACRITFPL LKPALRRGLT LAAATCVGEF 451 AATLFLSRPE WQTLTTLIYA YLGRAGEDNY ARAMVLTLLL AAFALGIFLL 501 LDGGEGGKQT ETL* Computer analysis of this amino acid sequence gave the following results: Homology with a predicted ORF from N.meninsitidis (strain Al ORF139 shows 94.7% identity over a 189aa overlap with an ORF (ORF139a) from strain A of N. meningitidis: 10 20 30 orf139.pep AWSAGESWRVLMESETWHAVWNTLRFSAAA I I II I I II I II I II I I I : I I I II II I I I I orfl39a QSVGEYVLLAFAAAVXSVCCLFXLLAIWKAWSAGESWRVLMESETWQAVWNTXRFSAAA 270 280 290 300 310 320 40 50 60 70 80 90 orf139.pep VYAAAVLGWYAAPARRSAWMRGLMFXPFMVSPVCVSAGVLLLYPQWTASLPLLLAMYAL I II I I II I II II I II I I I I II I I II I I II II I II II I II II II I II I I II II I II II orfl39a VYAAAVLGWYAAAARRSAWMRGLMFLPFMVSPVCVSAGVLLLXPQWTASLPLLLAMYAL 330 340 350 360 370 380 100 110 120 130 140 150 orf139.pep LAYPFVAKDVLSAWDALPPDYGRAAAGLGANGFQTACRITFPLLKPALRRGLTLAAATCV I I I M I II l I I I I I II II II I I II I II II I II I II II I I I I II I I I I II I II II I I I I I orfl39a LAYPFVAKDVLSAXDALPPDYGRAAAGLGANGFQTACRITFPLLKPALRRGLTLAAATCV 390 400 410 420 430 440 160 170 180 189 orf139. pep GEFAATLFLSRPEWQTLTTLIYAYLGRAGEDNYARAMVL I I II I II I II I I I I I I II I I I I II I I II I II I I II orfl39a GEFAATLFXSRXEWQTLTTLIYAYXGRAGXDNYARAMVLTLLLAAFALGXFLLLDGGEGG 450 460 470 480 490 500 The complete length ORF 139a nucleotide sequence <SEQ ID 577> is : 1 ATGGATGGAC GGCGTTGGGC GGTATGGGGT GCTTTTGCCC TGCTGCCTTC 51 GGCTTTTTTG GCGGCAATGG TCGTTGCGCC TTTGTGGGCG GTGGCGGCGT 101 ATGACGGTTT GGCGTGGCGC GCGGTGCTGT CGGATGCCTA TATGCTCAAA 151 CGTTTGGCGT GGACGGTATT TCAGGCAGCG GCAACCTGTG TGCTGGTGCT 201 GCCTTTGGGC GTGCCTGTCG CGTGGGTGCT GGCGCGGCTG GCGTTTCCGG 251 GGCGGGCTTT GGTGCTGCGC CTGCTGATGC TGCCTTTTGT GATGCCCACG 301 TTGGTGGCGG GCGTGGGCGT GCTGGCTCTG TTCGGGGCGG ACGGCCTGTN 351 GTGGCGCGGC TGGCAGGATA CGCCGTATCT GTTGTTGTAC GGCAATGTGT 401 TTTTTNACCT TCCTGTGTTG GTCAGGGCGG CATATCAGGG GTTTGTGCAA 451 GTGCCTGCGG CACGGCTTCA GACGGCACNG ACATTGGGCG CGGGGGCGTG 501 GCGGCGGTTT TGGGACATTG AAATGCCCGT TTTGCGCCCG TGGCTTGCCG 551 GCGGCGTGTG CCTTGTCTTC CTGTATTGTT TTTCGGGGTT CGGGCTGGCA 601 TTGCTGCTGG GCGGCAGCCG TTATGCCACG GTCGAAGTGG AAATTTACCA 651 GTTGGTCATG TTCGAACTCG ATATGGCGGT TGCTTCGGTG CTNGTGTGGC 701 TGGTGTNGGG GGTAACNGCG GCGGCAGGGT TGCTGTATGC GTGGTTCGGC 751 AGGCGCGCGG TTTCGGATAA GGCNGTTTCC CCTGTGATGC CGTCGCCGCC 801 GCAGTCGGTC GGGGAATATG TGCTNCTGGC GTTTGCGGCG GCGGTGTNGT 851 CTGTGTGCTG CCTGTTTCNT TTGTTGGCAA TTGTTGTGAA AGCGTGGTCG 901 GCCGGCGAAT CGTGGCGTGT GTTAATGGAA AGTGAAACGT GGCAGGCGGT 951 GTGGAATACT NTGCGCTTCT CGGCGGCGGC GGTGTATGCG GCGGCGGTTT 1001 TGGGTGTGGT GTATGCGGCG GCGGCGCGGC GGTCGGCGTG GATGCGCGGG 1051 CTGATGTTTT TGCCGTTTAT GGTGTCGCCG GTTTGTGTTT CGGCGGGCGT 1101 GCTGCTGCTT NATCCGCAGT GGACGGCTTC GTTGCCGCTG CTGCTGGCGA 1151 TGTATGCGCT GCTGGCGTAT CCGTTTGTGG CAAAAGATGT TTTATCAGCC 1201 TGNGATGCAC TGCCGCCGGA TTACGGCAGG GCGGCGGCGG GTTTGGGTGC 1251 AAACGGCTTT CAGACGGCAT GCCGCATCAC GTTCCCCCTC TTGAAACCGG 1301 CGTTGCGGCG CGGTCTGACT TTGGCGGCGG CAACCTGCGT GGGCGAATTT 1351 GCGGCAACCT TGTTCNTGTC GCGTCNCGAG TGGCAGACGC TGACGACTTT 1401 GATTTATGCC TATNTGGGAC GCGCGGGTGA NGATAATTAC GCGCGGGCGA 1451 TGGTGCTGAC ATTGCTGTTG GCGGCGTTCG CGCTGGGTAT NTTCCTGCTG 1501 TTGGACGGCG GCGAAGGCGG AAAACGGACG GAAACGTTAT AA This encodes a protein having amino acid sequence <SEQ ID 578>: 1 MDGRRWAVWG AFALLPSAFL AAMWAPLWA VAAYDGLAWR AVLSDAYMLK 51 RLAWTVFQAA ATCVLVLPLG VPVAWVLARL AFPGRALVLR LLMLPFVMPT 101 LVAGVGVLAL FGADGLXWRG WQDTPYLLLY GNVFFXLPVL VRAAYQGFVQ 151 VPAARLQTAX TLGAGAWRRF WDIEMPVLRP WLAGGVCLVF LYCFSGFGLA 201 LLLGGSRYAT VEVEIYQLVM FELDMAVASV LVWLVXGVTA AAGLLYAWFG 251 RRAVSDKAVS PVMPSPPQSV GEYVLLAFAA AVXSVCCLFX LLAIWKAWS 301 AGESWRVLME SETWQAVWNT XRFSAAAVYA AAVLGWYAA AARRSAWMRG 351 LMFLPFMVSP VCVSAGVLLL XPQWTASLPL LLAMYALLAY PFVAKDVLSA 401 XDALPPDYGR AAAGLGANGF QTACRITFPL LKPALRRGLT LAAATCVGEF 451 AATLFXSRXE WQTLTTLIYA YXGRAGXDNY ARAMVLTLLL AAFALGXFLL 501 LDGGEGGKRT ETL* ORF139a and ORF139-1 show 96.5% homology over a 514aa overlap: orf139a . pep MDGRRWAVWGAFALLPSAFLAAMWAPLWAVAAYDGLAWRAVLSDAYMLKRLAWTVFQAA I I I I IM I I I I I I I I I II I I I: II I I I I II I II I I I I I I I I I II I I II I I II II I I I I II orf139-1 MDGRRWWWGAFALLPSAFLAVMVVAPLWAVAAYDGLAWRAVLSDAYMLKRLAWTVFQAA orf139 . pep ATCVLVLPLGVPVAWVLARLAFPGRALVLRLLMLPFVMPTLVAGVGVLALFGADGLXWRG I I I I II II II I I II I I I I II I I I II I II I II II I I II I II II I II I I I I I I II II I III orf139-1 ATCVLVLPLGVPVAWVLARLAFPGRALVLRLLMLPFVMPTLVAGVGVLALFGADGLLWRG orf139a . pep WQDTPYLLLYGNVFFXLPVLVRAAYQGFVQVPAARLQTAXTLGAGAWRRFWDIEMPVLRP I I I II II II II II I I I I I I I I II I II I II I II I I II I I I II II I I I I I II I II II II orf139-1 RQDTPYLLLYGNVFFNLPVLVRAAYQGFVQVPAARLQTARTLGAGAWRRFWDIEMPVLRP orf139a . pep WLAGGVCLVFLYCFSGFGLALLLGGSRYATVEVEIYQLVMFELDMAVASVLVWLVXGVTA I I II I I I I II II II I II II II I I I I II II I I I I I II I I I I I II I I I II I I II II I I I II orf139-1 WLAGGVCLVFLYCFSGFGLALLLGGSRYATVEVEIYQLVMFELDMAVASVLVWLVLGVTA orf139a . pep AAGLLYAWFGRRAVSDKAVSPVMPSPPQSVGEYVLLAFAAAVXSVCCLFXLLAIWKAWS II II I I I I I I I II II II II I II I I I I I I II I II I I I I I II II II I I I I II I II I I I I I orf139-1 AAGLLYAWFGRRAVSDKAVSPVMPSPPQSVGEYVLLAFAAAVLSVCCLFPLLAIWKAWS orf139a . pep AGESWRVLMESETWQAVWNTXRFSAAAVYAAAVLGWYAAAARRSAWMRGLMFLPFMVSP II II I I I II II I I I I II I I I II I I I I I II I II I I II I II II I II II I I I I II II I I I I I orf139-1 AGESWRVLMESETWQAVWNTLRFSAAAVYAAAVLGWYAAAARRSAWMRGLMFLPFMVSP orf139a . pep VCVSAGVLLLXPQWTASLPLLLAMYALLAYPFVAKDVLSAXDALPPDYGRAAAGLGANGF I I I I I II I II I II I II I I II II I II I I II I II I I I I I I I I I I I I II I II I I II I I I I I orf139-1 VCVSAGVLLLYPQWTASLPLLLAMYALLAYPFVAKDVLSAWDALPPDYGRAAAGLGANGF orf139a . pep QTACRITFPLLKPALRRGLTLAAATCVGEFAATLFXSRXEWQTLTTLIYAYXGRAGXDNY I I I I I II I I I II II I I I I I II I I I II I I II I I I I I II I II I I I I II I II I II I III orf139-1 QTACRITFPLLKPALRRGLTLAAATCVGEFAATLFLSRPEWQTLTTLIYAYLGRAGEDNY orf139a . pep ARAMVLTLLLAAFALGXFLLLDGGEGGKRTETLX II II I I I II II I I I II I II I I I II I I I : I II I I orf139-1 ARAMVLTLLLAAFALGIFLLLDGGEGGKQTETLX Homology with a predicted ORF from N.sonorrhoeae ORF139 shows 95.2% identity over a 189aa overlap with a predicted ORF (ORF139ng) from N.gonorrhoeae: orf139. pep AWSAGESWRVLMESETWHAVWNTLRFSAAA 30 I II I I II I I I I II I I I: I I I I II I I I I II orfl39ng QSVGEYVLLAFSVAVLSVCCLFPLSAIWKAWSAGESRRVLMESETWQAVWNTLRFSAAA 327 orf139. pep VYAAAVLGWYAAPARRSAWMRGLMFXPFMVSPVCVSAGVLLLYPQWTASLPLLLAMYAL 90 I: I II II I II I I I III II I II I I I I I I II II I I I I I I I I II I II II II orfl39ng VFAAAVLGWYAAAARRLVWMRGLVFLPFMVSPVCVSAGVLLLYPGWTASLPLLLAMYAL 387 orf139. pep LAYPFVAKDVLSAWDALPPDYGRAAAGLGANGFQTACRITFPLLKPALRRGLTLAAATCV 150 I I II I I I II I II I II II II I II II II II II II II I I II II I I II II I I II I II I I I I I I I orfl39ng LAYPFVAKDVLSAWDALPPDYGRAAAGLGANGFQTACRITFPLLKPALRRGLTLAAATCV 447 orfl39.pep GEFAATLFLSRPEWQTLTTLIYAYLGRAGEDNYARAMVL 189 II II I II I II I I II II II I I I II II I I II II II I II I I I orfl39ng GEFAATLFLSRPEWQTLTTLIYAYLGRAGEDNYARAMVLTLLLSAFAVCIFLLLDNGEGG 507 The complete length ORF139ng nucleotide sequence <SEQ ID 579> is predicted to encode a protein having amino acid sequence <SEQ ID 580> 1 MDGRCWAVRG AFSLLPSAFL AVMWAPLWA VAAYDGLAWR AVLSDAYMLK 51 RLAWTVFQAA ATCVLVLPLG VPVAWVLARL AFPGRALVLR LLMLPFVMPT 101 LVAGVGVLAL FGADGLLWRG RQDTPYLLLY GNVFFNLPVL VRAAYQGFAQ 151 VPAARLQTAR TLGAGAWRPF WDIEMPVLRP WLAGGVCLVF LYCFSGFGLA 201 LLLGGSRYAT VEVEIYQLVM FELDMAGASA LVWLVLGVTA AAGLLYAWFG 251 RRAVSDKAVS PVMPSPPQSV GEYVLLAFSV AVLSVCCLFP LSAIWKAWS 301 AGESRRVLME SETWQAVWNT LRFSAAAVFA AAVLGWYAA AARRLVWMRG 351 LVFLPFMVSP VCVSAGVLLL YPGWTASLPL LLAMYALLAY PFVAKDVLSA 401 WDALPPDYGR AAAGLGANGF QTACRITFPL LKPALRRGLT LAAATCVGEF 451 AATLFLSRPE WQTLTTLIYA YLGRAGEDNY ARAMVLTLLL SAFAVCIFLL 501 LDNGEGGKRT ETL* Further work revealed a variant gonococcal DNA sequence <SEQ LD 581>: 1 ATGGATGGAC GGTGTTGGGC GGTACGGGGT GCTTTTTCCC TGCTGCCTTC 51 GGCTTTTTTG GCGGTAATGG TCGTTGCGCC TTTGTGGGCG GTGGCGGCGT 101 ATGACGGTTT GGCGTGGCGC GCGGTGCTGT CGGATGCCTA TATGCTCAAA 151 CGTTTGGCGT GGACGGTGTT TCAGGCGGCG GCAACCTGTG TGCTGGTGCT 201 GCCTTTGGGC GTGCCTGTCG CGTGGGTGCT GGCGCGGCTG GCGTTCCCGG 251 GGCGGGCTTT GGTGCTGCGC CTGCTGATGC TGCCGTTTGT GATGCCCACG 301 CTGGTGGCGG GCGTGGGCGT GCTGGCTCTG TTCGGGGCGG ACGGGCTGTT 351 GTGGCGCGGC CGGCAGGATA CGCCGTATCT GTTGTTGTAC GGCAATGTGT 401 TTTTCAACCT GCCCGTGTTG GTCAGGGCGG CGTATCAGGG GTTTGCTCAA 451 GTGCCTGCGG CACGGCTTCA GACGGCACGG ACGTTGGGCG CGGGGGCGTG 501 GCGGCGGTTT TGGGACATTG AAATGCCCGT TTTGCGCCCG TGGCTTGCCG 551 GCGGCGTGTG CCTTGTCTTC CTGTATTGTT TTTCGGGGTT CGGGCTGGCA 601 TTGCTGTTGG GCGGCAGCCG TTATGCCACG GTCGAAGTGG AAATTTACCA 651 GTTGGTTATG TTCGAACTCG ATATGGCGGG GGCTTCGGCG CTGGTGTGGC 701 TGGTGTTGGG GGTAACGGCG GCGGCAGGGT TGCTGTATGC GTGGTTCGGC 751 AGGCGCGCGG TTTCGGATAA GGCGGTTTCC CCCGTGATGC CGTCGCCGCC 801 GCAATCGGTG GGGGAATATG TATTGCTGGC ATTTTCGGTG GCGGTGTTGT 851 CCGTGTGCTG CCTGTTTCCT TTGTCGGCAA TTGTTGTGAA AGCGTGGTCG 901 GCCGGCGAAT CGCGGCGTGT GTTAATGGAA AGTGAAACGT GGCAGGCAGT 951 GTGGAATACt ttGCGCTTTT CGGCGGCGGC GGTGTTTGCG GCGGCGGTTT 1001 TGGGTGTGGT GTATGCGGCG GCGGCGCGGC GGCTGGTGTG GATGCGCGGA 1051 CTGGTGTTTT TACCGTTTAT GGTGTCGCCG GTTTGTGTTT CGGCGGGCGT 1101 GCTGCTGCTT TATCCGGGGT GGACGGCTTC GTTACCGCTG CTGCTGGCGA 1151 TGTATGCGCT GCTGGCGTAT CCGTTTGTGG CAAAAGATGT TTTATCGGCC 1201 TGGGATGCAC TGCCGCCGGA TTACGGCAGG GCGGCGGCAG GTTTGGGCGC 1251 AAACGGCTTT CAGACGGCAT GCCGTATCAC GTTCCCCCTC TTGAAACCGG 1301 CGTTGCGGCG CGGTCTGACT TTGGCGGCGG CGACGTGTGT GGGCGAATTT 1351 GCGGCAACCT TGTTCCTGTC GCGTCCGGAA TGGCAGACGT TGACGACTTT 1401 GATTTATGCC TATTTGGGGC GTGCGGGTGA GGACAATTAT GCGCGGGCAA 1451 TGGTGTTGAC ATTGCTGTTG TCGGCATTTG CGGTGTGCAT TTTCCTGCTG 1501 TTGGACAACG GCGAAGGCGg aaaACGGACG GAAACGTTAT AA This coπesponds to the .amino acid sequence <SEQ ID 582; ORF139ng-l>: 1 MDGRCWAVRG AFSLLPSAFL AVMWAPLWA VAAYDGLAWR AVLSDAYMLK 51 RLAWTVFQAA ATCVLVLPLG VPVAWVLARL AFPGRALVLR LLMLPFVMPT 101 LVAGVGVLAL FGADGLLWRG RQDTPYLLLY GNVFFNLPVL VRAAYQGFAQ 151 VPAARLQTAR TLGAGAWR. F WDIEMPVLRP WLAGGVCLVF LYCFSGFGLA 201 LLLGGSRYAT VEVEIYQLVM FELDMAGASA LVWLVLGVTA AAGLLYAWFG 251 RRAVSDKAVS PVMPSPPQSV GEYVLLAFSV AVLSVCCLFP LSAIWKAWS 301 AGESRRVLME SETWQAVWNT LRFSAAAVFA AAVLGWYAA AARRLVWMRG 351 LVFLPFMVSP VCVSAGVLLL YPGWTASLPL LLAMYALLAY PFVAKDVLSA 401 WDALPPDYGR AAAGLGANGF QTACRITFPL LKPALRRGLT LAAATCVGEF 451 AATLFLSRPE WQTLTTLIYA YLGRAGEDNY ARAMVLTLLL SAFAVCIFLL 501 LDNGEGGKRT ETL* ORF139ng-l and ORF139-1 show 95.9% identity over 513aa overlap: orf l39ng MDGRCWAVRGAFSLLPSAFLAVMWAPLWAVAAYDGLAWRAVLSDAYMLKRLAWTVFQAA I I I I M l I I I : I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I orf 139-1 MDGRRWWWGAFALLPSAFLAVMWAPLWAVAAYDGLAWRAVLSDAYMLKRLAWTVFQAA orfl39ng ATCVLVLPLGVPVAWVLARLAFPGRALVLRLLMLPFVMPTLVAGVGVLALFGADGLLWRG I II II II I II I I I II II I II I II I I I I I I II I I I II I II II II I I I II II I II II I I II I orf139-1 ATCVLVLPLGVPVAWVLARLAFPGRALVLRLLMLPFVMPTLVAGVGVLALFGADGLLWRG orfl39ng RQDTPYLLLYGNVFFNLPVLVRAAYQGFAQVPAARLQTARTLGAGAWRRFWDIEMPVLRP I I I II I II I I I I I I I I II I I II I II II I : I II I II I I I I I II I I I I I I I I II I I I I II I I orf139-1 RQDTPYLLLYGNVFFNLPVLVRAAYQGFVQVPAARLQTARTLGAGAWRRFWDIEMPVLRP orfl39ng WLAGGVCLVFLYCFSGFGLALLLGGSRYATVEVEIYQLVMFELDMAGASALVWLVLGVTA I I II II II I II I II II I II I I II I II I I I II I II II I I I I I I I II I I I: I I I I II I I II orf139-1 WLAGGVCLVFLYCFSGFGLALLLGGSRYATVEVEIYQLVMFELDMAVASVLVWLVLGVTA orfl39ng AAGLLYAWFGRRAVSDKAVSPVMPSPPQSVGEYVLLAFSVAVLSVCCLFPLSAIWKAWS I I II II I II II I I I I II I II I II I I I I I I I II I II I I I:: II II I I I II I I II I I I II I orfl39-l AAGLLYAWFGRRAVSDKAVSPVMPSPPQSVGEYVLLAFAAAVLSVCCLFPLLAIWKAWS orfl39ng AGESRRVLMESETWQAVWNTLRFSAAAVFAAAVLGWYAAAARRLVWMRGLVFLPFMVSP II I I II I II I II II I II I I II I I I I II : I I I II I I I II II I I I : I I I II : I I I I II I I orf139 AGESWRVLMESETWQAVWNTLRFSAAAVYAAAVLGWYAAAARRSAWMRGLMFLPFMVSP orfl39ng VCVSAGVLLLYPGWTASLPLLLAMYALLAYPFVAKDVLSAWDALPPDYGRAAAGLGANGF II I I I II I I I I I I II I II I I I I I II I I I II I I I II II I I II II I I I I II I II II I I II I orf139-1 VCVSAGVLLLYPQWTASLPLLLAMYALLAYPFVAKDVLSAWDALPPDYGRAAAGLGANGF orfl39ng QTACRITFPLLKPALRRGLTLAAATCVGEFAATLFLSRPEWQTLTTLIYAYLGRAGEDNY I I I II I II I II I II I I I I I I I II II II II I I II I I I I I I I I II II I I I I II I I II II I II orf139-1 QTACRITFPLLKPALRRGLTLAAATCVGEFAATLFLSRPEWQTLTTLIYAYLGRAGEDNY orfl39ng ARAMVLTLLLSAFAVCIFLLLDNGEGGKRTETL I I I I II I II I : II I : II I I II : I II I I : II I I orfl39-l ARAMVLTLLLAAFALGIFLLLDGGEGGKQTETL Based on the presence of a predicted binding-protein-dependent transport systems inner membrane component signature (underlined) in the gonococcal protein, it is predicted that the proteins from N. meningitidis and N.gonorrhoeae, and their epitopes, could be useful antigens for vaccines or diagnostics, or for raising antibodies. Example 70 The following partial DNA sequence was identified in N. meningitidis <SEQ ID 583>: 1 ATGGACGGCT GGACACAGAC GCTGTCCGCG CAAACCCTGT TGGGCATTTC 51 GGCGGCGGCA ATCATCCTCA TTCTGATTTT AATCGTCAGA TTCCGCATCC 101 ACGCGCTGCT GACACTGGTC ATCGTCAGCC TGCTGACGGC TTTGGCAACC 151 GGTTTGCCCA CAGGCAGCAT TGTCAAAGAC ATACTGGTCA AAAACTTCGG 201 CGGCACGCTC GGCGGCGTGG CGCTTCTGGT CGGCCTGGGC GCGATGCTCG 251 AACGTTTGGT C... This coπesponds to the .amino acid sequence <SEQ ID 584; ORF140>: 1 MDGWTQTLSA QTLLGISAAA IILILILIVR FRIHALLTLV IVSLLTALAT 51 GLPTGSIVKD ILVKNFGGTL GGVALLVGLG AMLERLV.. Further work revealed the complete nucleotide sequence <SEQ ID 585>: 1 ATGGACGGCT GGACACAGAC GCTGTCCGCG CAAACCCTGT TGGGCATTTC 51 GGCGGCGGCA ATCATCCTCA TTCTGATTTT AATCGTCAAA TTCCGCATCC 101 ACGCGCTGCT GACACTGGTC ATCGTCAGCC TGCTGACGGC TTTGGCAACC 151 GGTTTGCCCA CAGGCAGCAT TGTCAACGAC ATACTGGTCA AAAACTTCGG 201 CGGCACGCTC GGCGGCGTGG CGCTTCTGGT CGGCCTGGGC GCGATGCTCG 251 GACGTTTGGT CGAAACATCC GGCGGCGCAC AGTCGCTGGC GGACGCGCTG 301 ATCCGGATGT TCGGCGAAAA ACGCGCACCG TTCGCGCTGG GCGTTGCCTC 351 GCTGATTTTC GGCTTCCCGA TTTTCTTCGA TGCCGGACTA ATCGTCATGC 401 TGCCCATCGT GTTCGCCACC GCACGGCGCA TGAAACAGGA CGTACTGCCC 451 TTCGCGCTTG CCTCCATCGG CGCATTTTCC GTCATGCACG TCTTCCTGCC 501 GCCCCATCCG GGCCCGATTG CCGCTTCCGA ATTTTACGGC GCGAACATCG 551 GCCAAGTTTT GATTTTGGGT CTGCCGACCG CCTTCATCAC ATGGTATTTC 601 AGCGGCTATA TGCTCGGCAA AGTGTTGGGG CGCACCATCC ATGTTCCCGT 651 TCCCGAACTG CTCAGCGGCG GCACGCAAGA CAACGACCTG CCGAAAGAAC 701 CTGCCAAAGC AGGAACGGTC GTCGCCATCA TGCTGATTCC CATGCTGCTG 751 ATTTTCCTGA ATACCGGCGT ATCGGCCCTC ATCAGCGAAA AACTCGTAAG 801 TGCGGACGAA ACCTGGGTTC AGACGGCAAA AATAATCGGT TCGACACCGA 851 TCGCCCTTCT GATTTCCGTA TTGGTCGCAC TGTTTGTCTT GGGACGCAAA 901 CGCGGCGAAA GCGGCAGCGC GTTGGAAAAA ACCGTGGACG GCGCACTCGC 951 CCCCGTCTGT TCCGTGATTC TGATTACCGG CGCGGGCGGT ATGTTCGGCG 1001 GCGTTTTGCG CGCTTCCGGC ATCGGCAAGG CACTCGCCGA CAGCATGGCG 1051 GATTTGGGCA TTCCCGTCCT TTTGGGCTGT TTCCTTGTCG CCTTGGCACT 1101 GCGTATCGCG CAAGGTTCGG CAACCGTCGC CCTGACCACC GCCGCCGCGC 1151 TGATGGCTCC TGCCGTTGCC GCCGCCGGCT TTACCGACTG GCAGCTCGCC 1201 TGTATCGTAT TGGCAACGGC GGCAGGTTCG GTCGGTTGCA GCCACTTCAA 1251 CGACTCCGGC TTCTGGCTGG TCGGCCGTCT CTTGGACATG GACGTACCGA 1301 CCACGCTGAA AACCTGGACG GTCAACCAAA CCCTCATCGC ACTCATCGGC 1351 TTTGCCTTGT CCGCACTGCT GTTCGCCATC GTCTGA This coπesponds to the amino acid sequence <SEQ ID 586; ORF140-1>: 1 MDGWTQTLSA QTLLGISAAA IILILILIVK FRIHALLTLV IVSLLTALAT 51 GLPTGSIVND ILVKNFGGTL GGVALLVGLG AMLGRLVETS GGAQSLADAL 101 IRMFGEKRAP FALGVASLIF GFPIFFDAGL IVMLPIVFAT ARRMKQDVLP 151 FALASIGAFS VMHVFLPPHP GPIAASEFYG ANIGQVLILG LPTAFITWYF 201 SGYMLGKVLG RTIHVPVPEL LSGGTQDNDL PKEPAKAGTV VAIMLIPMLL 251 IFLNTGVSAL ISEKLVSADE TWVQTAKIIG STPIALLISV LVALFVLGRK 301 RGESGSALEK TVDGALAPVC SVILITGAGG MFGGVLRASG IGKALADSMA 351 DLGIPVLLGC FLVALALRIA QGSATVALTT AAALMAPAVA AAGFTDWQLA 401 CIVLATAAGS VGCSHFNDSG FWLVGRLLDM DVPTTLKTWT VNQTLIALIG 451 FALSALLFAI V* Computer analysis of this amino acid sequence gave the following results: Homology with a predicted ORF from N.meninsitidis (sfrain A) ORF140 shows 95.4% identity over a 87aa overlap with an ORF (ORFMOa) from strain A of N. meningitidis: 10 20 30 40 50 60 orf140. pep MDGWTQTLSAQTLLGISAAAIILILILIVRFRIHALLTLVIVSLLTALATGLPTGSIVKD I I I I I I I I II I II I I II II I II I II I II I : I II I II I II I I II II II II I II I I I I I I : I orfl40a MDGWTQTLSAQTLLGISAAAIILILILIVKFRIHALLTLVIVSLLTALATGLPTGSIVND 10 20 30 40 50 60 70 80 orf140. pep ILVKNFGGTLGGVALLVGLGAMLERLV : I II I I I II I I I I I I I I I I I I I I III orf140a VLVKNFGGTLGGVALLVGLGAMLGRLVETSGGAQSLADALIRMFGEKRAPFALGVASLIF 70 80 90 100 110 120 The complete length ORF 140a nucleotide sequence <SEQ ID 587> is: 1 ATGGACGGCT GGACACAGAC GCTGTCCGCG CAAACCCTGT TGGGCATTTC 51 GGCGGCGGCA ATCATCCTCA TTCTGATTTT AATCGTCAAA TTCCGCATCC 101 ACGCGCTGCT GACACTGGTC ATCGTCAGCC TGCTGACGGC TTTGGCAACC 151 GGTTTGCCCA CAGGCAGCAT TGTCAACGAC GTACTGGTCA AAAACTTCGG 201 CGGCACGCTC GGCGGCGTGG CGCTTCTGGT CGGCCTGGGC GCGATGCTCG 251 GACGTTTGGT CGAAACATCC GGCGGCGCAC AGTCGCTGGC GGACGCGCTG 301 ATCCGGATGT TCGGCGAAAA ACGCGCACCG TTCGCGCTGG GCGTTGCCTC 351 GCTGATTTTC GGCTTCCCGA TTTTCTTCGA TGCCGGACTA ATCGTCATGC 401 TGCCCATCGT GTTCGCCACC GCACGGCGCA TGAAACAGGA CGTACTGCCC 451 TTCGCGCTTG CCTCCATCGG CGCATTTTCC GTCATGCACG TCTTCCTGCC 501 GCCCCATCCG GGCCCGATTG CCGCTTCCGA ATTTTACGGC GCGAACATCG 551 GCCAAGTTTT GATTTTGGGT CTGCCGACCG CCTTCATCAC ATGGTATTTC 601 AGCGGCTATA TGCTCGGCAA AGTGTTGGGG CGCACCATCC ATGTTCCCGT 651 TCCCGAACTG CTCAGCGGCG GCACGCAAGA CAACGACCTG CCGAAAGAAC 701 CTGCCAAAGC AGGAACGGTC GTCGCCATCA TGCTGATTCC CATGCTGCTG 751 ATTTTCCTGA ATACCGGCGT ATCGGCCCTC ATCAGCGAAA AACTCGTAAG 801 TGCGGACGAA ACCTGGGTTC AGACGGCAAA AATAATCGGT TCGACACCGA 851 TCGCCCTTCT GATTTCCGTA TTGGTCGCAC TGTTTGTCTT GGGACGCAAA 901 CGCGGCGAAA GCGGCAGCGC GTTGGAAAAA ACCGTGGACG GCGCACTCGC 951 CCCCGTCTGT TCCGTGATTC TGATTACCGG CGCGGGCGGT ATGTTCGGCG 1001 GCGTTTTGCG CGCTTCCGGC ATCGGCAAGG CACTCGCCGA CAGCATGGCG 1051 GATTTGGGCA TTCCCGTCCT TTTGGGCTGT TTCCTTGTCG CCTTGGCACT 1101 GCGTATCGCG CAAGGTTCGG CAACCGTCGC CCTGACCACC GCCGCCGCGC 1151 TGATGGCTCC TGCCGTTGCC GCCGCCGGCT TTACCGACTG GCAGCTCGCC 1201 TGTATCGTAT TGGCAACGGC GGCAGGTTCG GTCGGTTGCA GCCACTTCAA 1251 CGACTCCGGC TTCTGGCTGG TCGGCCGCCT CTTGGACATG GACGTACCGA 1301 CCACGCTGAA AACCTGGACG GTCAACCAAA CCCTCATCGC ACTCATCGGC 1351 TTTGCCTTGT CCGCACTGCT GTTCGCCATC GTCTGA This encodes a protein having amino acid sequence <SEQ ID 588>: 1 MDGWTQTLSA QTLLGISAAA IILILILIVK FRIHALLTLV IVSLLTALAT 51 GLPTGSIVND VLVKNFGGTL GGVALLVGLG AMLGRLVETS GGAQSLADAL 101 IRMFGEKRAP FALGVASLIF GFPIFFDAGL IVMLPIVFAT ARRMKQDVLP 151 FALASIGAFS VMHVFLPPHP GPIAASEFYG ANIGQVLILG LPTAFITWYF 201 SGYMLGKVLG RTIHVPVPEL LSGGTQDNDL PKEPAKAGTV VAIMLIPMLL 251 IFLNTGVSAL ISEKLVSADE TWVQTAKIIG STPIALLISV LVALFVLGRK 301 RGESGSALEK TVDGALAPVC SVILITGAGG MFGGVLRASG IGKALADSMA 351 DLGIPVLLGC FLVALALRIA QGSATVALTT AAALMAPAVA AAGFTDWQLA 401 CIVLATAAGS VGCSHFNDSG FWLVGRLLDM DVPTTLKTWT VNQTLIALIG 451 FALSALLFAI V* ORF140a and ORF140-1 show 99.8% identity over a 461aa overlap: orf140-1.pep MDGWTQTLSAQTLLGISAAAIILILILIVKFRIHALLTLVIVSLLTALATGLPTGSIVND 60 I I I I I I I I I I I I I I II I II I I I I II I II I I I II I I II I II I I I II I I I II II I I I I II II orfl40a MDGWTQTLSAQTLLGISAAAIILILILIVKFRIHALLTLVIVSLLTALATGLPTGSIVND 60 orf140-1.pep ILVKNFGGTLGGVALLVGLGAMLGRLVETSGGAQSLADALIRMFGEKRAPFALGVASLIF 120 : I I II II I II I I II I II I II I II I II I II II I II II I II I II I I I I I II I I II I I II I II orfl40a VLVKNFGGTLGGVALLVGLGAMLGRLVETSGGAQSLADALIRMFGEKRAPFALGVASLIF 120 orf140-1.pep GFPIFFDAGLIVMLPIVFATARRMKQDVLPFALASIGAFSVMHVFLPPHPGPIAASEFYG 180 II I II I I I I I I I II I I I I I I I II I II I II II I II II I I I I II I I I II I I I I II II II I II orfl40a GFPIFFDAGLIVMLPIVFATARRMKQDVLPFALASIGAFSVMHVFLPPHPGPIAASEFYG 810 orf140-1.pep ANIGQVLILGLPTAFITWYFSGYMLGKVLGRTIHVPVPELLSGGTQDNDLPKEPAKAGTV 240 I I I I I I I I I I I I II I I I I I I I I I I II I II II I II I I I I I I I I I I I I I I I I I I II I II I I I orfl40a ANIGQVLILGLPTAFITWYFSGYMLGKVLGRTIHVPVPELLSGGTQDNDLPKEPAKAGTV 240 orf140-1.pep VAIMLIPMLLIFLNTGVSALISEKLVSADETWVQTAKIIGSTPIALLISVLVALFVLGRK 300 I I I I I I I I I I I I II I I I I I I I II I I I I II I II I II I II I II II I I I I I I II I I II II I I I orfl40a VAIMLIPMLLIFLNTGVSALISEKLVSADETWVQTAKIIGSTPIALLISVLVALFVLGRK 300 orf140-1. pep RGESGSALEKTVDGALAPVCSVILITGAGGMFGGVLRASGIGKALADSMADLGIPVLLGC 360 I I I I I I II I I I I I I II I II I I I I I II I II I II II I I II I II I I I I I I I I II I II II I I I I orfl40a RGESGSALEKTVDGALAPVCSVILITGAGGMFGGVLRASGIGKALADSMADLGIPVLLGC 360 orfl40-l. pep FLVALALRIAQGSATVALTTAAALMAPAVAAAGFTDWQLACIVLATAAGSVGCSHFNDSG 420 I II I I I I II II II I II I II I I I I I II I II I II II I I II I II II I I I I I I I I I I II II I I I orfl40a FLVALALRIAQGSATVALTTAAALMAPAVAAAGFTDWQLACIVLATAAGSVGCSHFNDSG 420 orf140-1. pep FWLVGRLLDMDVPTTLKTWTVNQTLIALIGFALSALLFAIV 461 II I II I I I II II I II I II I I I I I II I I I I II I II II II I II orf140a FWLVGRLLDMDVPTTLKTWTVNQTLIALIGFALSALLFAIV 461 Homology with a predicted ORF from N.sonorrhoeae ORF140 shows 92% identity over a 87aa overlap with a predicted ORF (ORF140ng) from N.gonorrhoeae: orfl40.pep MDGWTQTLSAQTLLGISAAAIILILILIVRFRIHALLTLVIVSLLTALATGLPTGSIVKD 60 III I II II II II I II I I I I I II I II I II : I I I : I I I I I I I : II II II I I I II I I I I I : I orfl40ng MDGRTQTLSAQTLLGISAAAIILILILIVKFRIRALLTLVIASLLTALATGLPTGSIVND 60 orf140. pep ILVKNFGGTLGGVALLVGLGAMLERLV 87 : I I I I II II I I II II II I I II I I III orf14Ong VLVKNFGGTLGGVALLVGLGAMLGRLVETSGGAQSLADALIRMFGEKRAPFAPGVASLIF 120 The complete length ORF140ng nucleotide sequence <SEQ ID 589> was predicted to encode a protein having amino acid sequence <SEQ ED 590>: 1 MDGRTOTLSA QTLLGISAAA IILILILIVK FRIRALLTLV IASLLTALAT 51 GLPTGSIVND VLVKNFGGTL GGVALLVGLG AMLGRLVETS GGAQSLADAL 101 IRMFGEKRAP FAPGVASLIF GFPIFFDAGL IVMLPIVFAT ARRMKQDVLP 151 FALASVGAFS VMHVFLPPHP GPIAASEFYG ANIGQVLILG LPTAFITWYF 201 SGYMLGKVLG RAIHVPVPEL LSGGTQDSDP PKEPAKAGTV VAVMLIPMLL 251 IFLNTGVSAL ISEKLVSADE TWVQTAKMIG STPVALLISV LAALLVLGRK 301 RGESGSTLEK TVDGALAPAC SVILITGAGG MFGGVLRASG IGKALADSMA 351 DLGIPVLLGC FLVALALRIA QGSATVALTT AAALMAPAVA AAGFTDWQLA 401 CIVLATAAGS VGCSHFNDSG FWLVGRLSDM DVPTTLKTWT VNQTLIAFIG 451 FALSALLFAI V* Further work revealed a variant gonococcal DNA sequence <SEQ ED 591>: 1 ATGGACGGCC GGACACAGAC GCTGTCCGCG CAAACCTTGT TGGGCATTTC 51 GGCGGCGGCA ATCATCCTCA TTCTGATTTT AATCGTCAAA TTCCGCATCC 101 GCGCGCTGCT GACACTGGTC ATCGCCAGCC TGCTGACGGC TTTGGCAACC 151 GGTTTGCCCA CAGGCAGCAT CGTCAACGAC GTACTGGTCA AAAACTTCGG 201 CGGCACGCTC GGCGGCGTGG CGCTTCTGGT CGGTCTGGGC GCAATGCTCG 251 GACGTTTGGT AGAAACATCC GGCGGCGCAC AGTCGCTGGC GGACGCGCTG 301 ATCCGGATGT TCGGCGAAAA ACGCGCACCG TTCGCTCCGG GCGTTGCCTC 351 GCTGATTTTC GGCTTCCCGA TTTTCTTCGA TGCCGGACTA ATCGTCATGC 401 TGCCCATCGT ATTCGCCACC GCACGGCGCA TGAAACAGGA CGTACTGCCC 451 TTCGCGCTTG CCTCCGTCGG CGCATTTTCC GTCATGCACG TCTTCCTGCC 501 GCCCCATCCG GGCCCGATTG CCGCTTCCGA ATTTTACGGC GCGAACATCG 551 GCCAGGTTTT GATTTTGGGT CTGCCGACCG CCTTCATCAC ATGGTATTTC 601 AGCGGCTATA TGCTCGGCAA AGTGTTGGGG CGCGCCATCC ATGTTCCCGT 651 TCCCGAACTG CTCAGCGGCG GCACGCAAGA CAGCGACCCG CCGAAAGAAC 701 CTGCCAAAGC AGGAACGGTC GTCGCCGTCA TGCTGATTCC CATGCTGCTG 751 ATTTTCCTGA ATACCGGCGT ATCAGCCCTC ATCAGCGAAA AACTCGTAAG 801 TGCGGACGAA ACTTGGGTTC AGACGGCAAA AATGATCGGT TCGACACCTG 851 TCGCCCTTCT GATTTCCGTA TTGGCCGCAC TGTTGGTCTT GGGACGCAAA 901 CGCGGCGAAA GCGGCAGCAC GTTGGAAAAA ACCGTGGACG GCGCACTCGC 951 CCCCGCCTGT TCCGTGATTC TGATTACCGG CGCGGGCGGT ATGTTCGGCG 1001 GCGTTTTGCG CGCTTCCGGC ATCGGCAAGG CACTCGCCGA CAGCATGGCG 1051 GATTTGGGCA TTCCCGTCCT TTTGGGCTGC TTCCTTGTCG CCTTGGCACT 1101 GCGTATCGCG CAAGGTTCGG CAACCGTCGC CCTGACCACA GCCGCCGCGC 1151 TGATGGCTCC TGCCGTTGCC GCCGCCGGCT TTACCGACTG GCAGCTCGCC 1201 TGTATCGTAT TGGCAACGGC GGCAGGTTCG GTCGGTTGCA GCCACTTCAA 1251 CGACTCCGGC TTCTGGCTGG TCGGCCGCCT CTTGGATATG GACGTACCGA 1301 CCACGCTGAA AACCTGGACG GTCAACCAAA CCCTCATCGC ATTCATCGGC 1351 TTTGCCTTGT CCGCACTGCT GTTTGCCATC GTCTGA This coπesponds to the amino acid sequence <SEQ ID 592; ORF140ng-l>: 1 MDGRTQTLSA QTLLGISAAA IILILILIVK FRIRALLTLV IASLLTALAT 51 GLPTGSIVND VLVKNFGGTL GGVALLVGLG AMLGRLVETS GGAQSLADAL 101 IRMFGEKRAP FAPGVASLIF GFPIFFDAGL IVMLPIVFAT ARRMKQDVLP 151 FALASVGAFS VMHVFLPPHP GPIAASEFYG ANIGQVLILG LPTAFITWYF 201 SGYMLGKVLG RAIHVPVPEL LSGGTQDSDP PKEPAKAGTV VAVMLIPMLL 251 IFLNTGVSAL ISEKLVSADE TWVQTAKMIG STPVALLISV LAALLVLGRK 301 RGESGSTLEK TVDGALAPAC SVILITGAGG MFGGVLRASG IGKALADSMA 351 DLGIPVLLGC FLVALALRIA QGSATVALTT AAALMAPAVA AAGFTDWQLA 401 CIVLATAAGS VGCSHFNDSG FWLVGRLiDM DVPTTLKTWT VNQTLIAFIG 451 FALSALLFAI V* ORF140ng-l and ORF140-1 show 96.3% identity over 461aa overlap: orfl 40ng-l . pep MDGRTQTLSAQTLLGISAAAIILILILIVKFRIRALLTLVIASLLTALATGLPTGSIVND I I I I I I I I I I I M I I I I I I I I I I I I I I I I I I M M I I I I M I I I I I I I I I I I I I I I I I I orfl40-l MDGWTQTLSAQTLLGISAAAIILILILIVKFRIHALLTLVIVSLLTALATGLPTGSIVND orf140ng-l . pep VLVKNFGGTLGGVALLVGLGAMLGRLVETSGGAQSLADALIRMFGEKRAPFAPGVASLIF : I I II II I II II II I I I I II I II II I II I I I I II II I II I II I I I II I I II I II I II I I orf140-1 ILVKNFGGTLGGVALLVGLGAMLGRLVETSGGAQSLADALIRMFGEKRAPFALGVASLIF orfl40ng-l.pep GFPIFFDAGLIVMLPIVFATARRMKQDVLPFALASVGAFSVMHVFLPPHPGPIAASEFYG II I I I II I II I I II I II I II I II II I I I I I I I II I : I II I I I I II II I I II I I I I I I II I orf140-1 GFPIFFDAGLIVMLPIVFATARRMKQDVLPFALASIGAFSVMHVFLPPHPGPIAASEFYG orfl40ng-l.pep ANIGQVLILGLPTAFITWYFSGYMLGKVLGRAIHVPVPELLSGGTQDSDPPKEPAKAGTV I I II II I I I I I I I II II I II I II II I I I I I I : II II I I I I I I I I I I I : I I I I I I I I I II orf140-1 ANIGQVLILGLPTAFITWYFSGYMLGKVLGRTIHVPVPELLSGGTQDNDLPKEPAKAGTV orf140ng-l .pep VAVMLIPMLLIFLNTGVSALISEKLVSADETWVQTAKMIGSTPVALLISVLAALLVLGRK II : I I I I I I I I I II II II I II II I II I I II II I I I I I : I I I II : I I I II I I : II : II II I or5140-1 VAIMLIPMLLIFLNTGVSALISEKLVSADETWVQTAKIIGSTPIALLISVLVALFVLGRK orfl40ng-l.pep RGESGSTLEKTVDGALAPACSVILITGAGGMFGGVLRASGIGKALADSMADLGIPVLLGC I II II I : I I I I II II II I : I II I II I II II II I I I I II II I II I I I II I I I II I I I II II orfl40-l RGESGSALEKTVDGALAPVCSVILITGAGGMFGGVLRASGIGKALADSMADLGIPVLLGC orfl40ng-l.pep FLVALALRIAQGSATVALTTAAALMAPAVAAAGFTDWQLACIVLATAAGSVGCSHFNDSG II I II I II I II II II II I II I I I II I II I I I I I I I I II I I I II I I I II I II I I I I I I II I orfl40-l FLVALALRIAQGSATVALTTAAALMAPAVAAAGFTDWQLACIVLATAAGSVGCSHFNDSG orf140ng-l .pep FWLVGRLLDMDVPTTLKTWTVNQTLIAFIGFALSALLFAIV II I I I II I II II I I I II II I I I I II I I : I I II I I I I I I II I orf140-1 FWLVGRLLDMDVPTTLKTWTVNQTLIALIGFALSALLFAIV Furthermore, ORF140ng-l is homologous to an E.coli protein: gi|882633 (U29579) ORF_o454 [Escherichia coli] >gi|1789097 (AE000358) o454; This 454 aa ORF is 34% identical (9 gaps) to 444 residues of an approx. 456 aa protein GNTP_BACLI SW: P46832 [Escherichia coli] Length = 454 Score = 210 bits (529), Expect = le-53 Identities = 130/384 (33%), Positives = 194/384 (49%), Gaps = 19/384 (4%) Query: 88 ETSGGAQSLADALIRMFGEKRAPFAPGVASLIFGFPIFFDAGLIVMLPIVFATARRMKQD 147 E SGGA+SLA+ R G+KR A +A+ G P+FFD G I++ PI++ A+ K Sbjct: 80 EHSGGAESLANYFSRKLGDKRTIAALTLAAFFLGIPVFFDVGFIILAPIIYGFAKVAKIS 139 Query: 148 VLPFALASVGAFSVMHVFLPPHPGPIAASEFYGANIGQVLILGLPTAFITWYFSGYMLGK 207 L F L G +HV +PPHPGP+AA+ A+IG + I+G+ + 1 GY K Sbjct: 140 PLKFGLPVAGIMLTVHVAVPPHPGPVAAAGLLHADIGWLTIIGIAIS-IPVGWGYFAAK 198 Query: 208 VLGRAIHVPVPELL SGGTQDSDPPKEPAKAGTWAVMLIPMLLIFLNTGV 257 ++ + + E+L G T+ SD P A V ++++IP+ +1 T Sbjct: 199 IINKRQYAMSVEVLEQMQLAPASEEGATKLSDKINPPGVA-LVTSLIVIPIAIIMAGT-- 255 Query: 258 SALISEKLVSADETWVQTAKMIGSTPXXXXXXXXXXXXXXGRKRGESGSTLEKTVDGALA 317 +S L+ + T ++IGS +RG S + AL Sbjct: 256 VSATLMPPSHPLLGTLQLIGSPMVALMIALVLAFWLLALRRGWSLQHTSDIMGSALP 312 Query: 318 PACSVILITGAGGMFGGVLRASGIGKALADSMADLGIPVLLGCFLVALALRIAQGSXXXX 377 A VIL+TGAGG+FG VL SG+GKALA+ + + +P+L F+++LALR +QGΞ Sbjct: 313 TAAWILVTGAGGVFGKVLVESGVGKALANMLQMIDLPLLPAAFIISLALRASQGS—AT 370 Query: 378 XXXXXXXXXXXXXXXGFTDWQLACIVLATAAGSVGCSHFNDSGFWLVGRLLDMDVPTTLK 437 G Q + LA G +G SH NDSGFW+V + L + V LK Sbjct: 371 VAILTTGGLLSEAVMGLNPIQCVLVTLAACFGGLGASHINDSGFWIVTKYLGLSVADGLK 430 Query: 438 TWTVNQTLIAFIGFALSALLFAIV 461 TWTV T++ F GF ++ ++A++ Sbjct: 431 TWTVLTTILGFTGFLITWCVWAVI 454 Based on this analysis, including the identification of the presence of a putative leader sequence (double-underlined) and several putative transmembrane domains (single-underlined) in the gonococcal protein, it is predicted that the proteins from N. meningitidis and N.gonorrhoeae, and their epitopes, could be useful antigens for vaccines or diagnostics, or for raising antibodies. Example 71 The following partial DNA sequence was identified in N. meningitidis <SEQ ID 593>: 1 .. GATTTCGGCA TATCGCCCGT GTATCTTTGG GTTGCCGCCG CGTTCAAACA 51 TTTGCTGTCG CCGTGGGCTG CCGACTCATA CGATGTCGCA CGCTTTGCAG 101 GCGTATTTTT TGCCGTTATC GGACTGACTT CCTGCGGCTT TGCCGGTTTC 151 AACTTTTTGG GCAGACACCA CGGGCGCAC . GTCGTCCTGA TTCTCATCGG 201 CTGTATCGGG CTGATTCCAG TTGCCCATTT CCTCAACCCC GCTGCCGCCG 251 CCTTTGCCGC CGCCGGACTG GTGCTGCACG GTTATTCTTT GGCTCGCCGG 301 CGCGTGATTG CCGCCTCTTT TCTGCTCGGT ACGGGCTGGA CGCTGATGTC 351 GTTGGCAGCA GCTTATCCGG CAGCATTTGC CCTGATGCTG CCCTTGCCCG 401 TACTGATGTT TTTCCGTCCG .. This coπesponds to the amino acid sequence <SEQ ID 594; ORF141>: 1 ..DFGISPVYLW VAAAFKHLLS PWAADSYDVA RFAGVFFAVI GLTSCGFAGF 51 NFLGRHHGRX WLILIGCIG LIPVAHFLNP AAAAFAAAGL VLHGYSLARR 101 RVIAASFLLG TGWTLMSLAA AYPAAFALML PLPVLMFFRP .. Further work revealed the complete nucleotide sequence <SEQ ID 595>: 1 ATGCTGACCT ATACCCCGCC CGATGCCCGC CCGCCCGCCA AAACCCACGA 51 AAAGCCGTGG CTGCTGCTGT TGATGGCGTT TGCCTGGTTG TGGCCCGGCG 101 TGTTTTCCCA CGATTTGTGG AATCCTGACG AACCTGCCGT CTATACCGCC 151 GTCGAAGCAC TGGCAGGCAG CCCCACCCCC TTGGTTGCCC ATCTGTTCGG 201 TCAAACCGAT TTCGGCATAC CGCCCGTGTA TCTTTGGGTT GCCGCCGCGT 251 TCAAACATTT GCTGTCGCCG TGGGCTGCCG ACTCATACGA TGCCGCACGC 301 TTTGCAGGCG TATTTTTTGC CGTTATCGGA CTGACTTCCT GCGGCTTTGC 351 CGGTTTCAAC TTTTTGGGCA GACACCACGG GCGCAgCGTC GTCCTGATTC 401 TCATCGGCTG TATCGGGCTG ATTCCAGTTG CCCATTTCCT CAACCCCGCT 451 GCCGCCGCCT TTGCCGCCGC CGGACTGGTG CTGCACGGTT ATTCTTTGGC 501 TCGCCGGCGC GTGATTGCCG CCTCTTTTCT GCTCGGTACG GGCTGGACGC 551 TGATGTCGTT GGCAGCAGCT TATCCGGCAG CATTTGCCCT GATGCTGCCC 601 TTGCCCGTAC TGATGTTTTT CCGTCCGTGG CAAAGCAGGC GTTTGATGTT 651 GACGGCAGTC GCCTCACTTG CCTTTGCCCT GCCGCTTATG ACCGTTTACC 701 CGCTGCTCTT GGCAAAAACG CAGCCCGCGC TGTTCGCGCA ATGGCTCGAC 751 TATCACGTTT TCGGTACGTT CGGCGGCGTG CGGCACGTTC AGACGGCATT 801 CAGTTTGTTT TACTATCTGA AAAACCTGCT TTGGTTTGCA TTGCCCGCGC 851 TGCCGCTGGC GGTTTGGACG GTTTGCCGCA CGCGCCTGTT TTCGACCGAC 901 TGGGGGATTT TGGGCGTCGT CTGGATGCTT GCCGTTTTGG TGCTGCTTGC 951 CGTCAATCCG CAGCGTTTTC AGGATAACCT CGTCTGGCTG CTTCCGCCGC 1001 TTGCCCTGTT CGGCGCGGCG CAACTGGACA GCCTGAGGCG CGGCGCGGCG 1051 GCGTTTGTCA ACTGGTTCGG CATTATGGCG TTCGGACTGT TTGCCGTGTT 1101 CCTGTGGACG GGCTTTTTCG CCATGAATTA CGGCTGGCCC GCCAAGCTTG 1151 CCGAACGCGC CGCCTATTTC AGCCCGTATT ATGTTCCTGA TATCGATCCC 1201 ATTCCGATGG CGGTTGCCGT ACTGTTCACA CCCTTGTGGC TGTGGGCGAT 1251 TACCCGGAAA AACATACGCG GCAGGCAGGC GGTTACCAAC TGGGCGGCAG 1301 GCGTTACCCT GACCTGGGCT TTGCTGATGA CGCTGTTCCT GCCGTGGCTG 1351 GACGCGGCGA AAAGCCACGC GCCGGTCGTC CGGAGTATGG AGGCATCGCT 1401 TTCCCCGGAA TTGAAACGGG AGCTTTCAGA CGGCATCGAG TGTATCGGCA 1451 TAGGCGGCGG CGACCTGCAC ACGCGGATTG TTTGGACGCA GTACGGCACA 1501 TTGCCGCACC GCGTCGGCGA TGTACAATGC CGCTACCGCA TCGTCCTCCT 1551 GCCCCAAAAT GCGGATGCGC CGCAAGGCTG GCAGACGGTT TGGCAGGGTG 1601 CGCGTCCGCG CAACAAAGAC AGTAAGTTCG CACTGATACG GAAAATCGGG 1651 GAAAATATAT AA This coπesponds to the amino acid sequence <SEQ ID 596; ORF141-l>: 1 MLTYTPPDAR PPAKTHEKPW LLLLMAFAWL WPGVFSHDLW NPDEPAVYTA 51 VEALAGSPTP LVAHLFGQTD FGIPPVYLWV AAAFKHLLSP WAADSYDAAR 101 FAGVFFAVIG LTSCGFAGFN FLGRHHGRSV VLILIGCIGL IPVAHFLNPA 151 AAAFAAAGLV LHGYSLARRR VIAASFLLGT GWTLMSLAAA YPAAFALMLP 201 LPVLMFFRPW QSRRLMLTAV ASLAFALPLM TVYPLLLAKT QPALFAQWLD 251 YHVFGTFGGV RHVQTAFSLF YYLKNLLWFA LPALPLAVWT VCRTRLFSTD 301 WGILGWWML AVLVLLAVNP QRFQDNLVWL LPPLALFGAA QLDSLRRGAA 351 AFVNWFGIMA FGLFAVFLWT GFFAMNYGWP AKLAERAAYF SPYYVPDIDP 401 IPMAVAVLFT PLWLWAITRK NIRGRQAVTN WAAGVTLTWA LLMTLFLPWL 451 DAAKSHAPVV RSMEASLSPE LKRELSDGIE CIGIGGGDLH TRIVWTQYGT 501 LPHRVGDVQC RYRIVLLPQN ADAPQGWQTV WQGARPRNKD SKFALIRKIG 551 ENI* Computer analysis of this amino acid sequence gave the following results: Homology with a predicted ORF from N.meninsitidis (sfrain A) ORF141 shows 95.0% identity over a 140aa overlap with an ORF (ORF141a) from strain A of N. meningitidis: 10 20 30 orf141. pep DFGISPVYLWVAAAFKHLLSPWAADSYDVA II I I I II I I I I I I II I I I I I II I I I I: I orfl41a WNPDEPAVYTAVEALAGSPTPLVAHLFGQIDFGIPPVYLWVAAAFKHLLSPWAADPYDAA 40 50 60 70 80 90 40 50 60 70 80 90 orf141.pep RFAGVFFAVIGLTSCGFAGFNFLGRHHGRXWLILIGCIGLIPVAHFLNPAAAAFAAAGL M I I I I M I : M I I M M M M I I I I M I I M I II I II II II : : I I I II I I II II II I I orfl41a RFAGVFFAWGLTSCGFAGFNFLGRHHGRSWLILIGCIGLIPTVHFLNPAAAAFAAAGL 100 110 120 130 140 150 100 110 120 130 140 orf141. pep VLHGYSLARRRVIAASFLLGTGWTLMSLAAAYPAAFALMLPLPVLMFFRP II II II I II I I I II I I I II I I II I II I II I II I I I I II I I II II I II I I I orfl41a VLHGYSLARRRVIAASFLLGTGWTLMSLAAAYPAAFALMLPLPVLMFFRPWQSRRLMLTA 160 170 180 190 200 210 orf14 la VASLAFALPLMTVYPLLLAKTQPALFAQWLDDHVFGTFGGVRHIQTAFSLFYYLKNLLWF 220 230 240 250 260 270 The complete length ORF141 a nucleotide sequence <SEQ ID 597> is: 1 ATGCTGACCT ATACCCCGCC CGATGCCCGC CCGCCCGCCA AAACCCACGA 51 AAAGCCGTGG CTGTTGCTGT TGATGGCGTT TGCCTGGTTG TGGCCCGGCG 101 TGTTTTCCCA CGATTTGTGG AATCCTGACG AACCTGCCGT CTATACCGCC 151 GTCGAAGCAC TGGCAGGCAG CCCCACCCCT TTGGTTGCCC ATCTGTTCGG 201 TCAAATCGAT TTCGGCATAC CGCCCGTGTA TCTTTGGGTT GCCGCCGCGT 251 TCAAACATTT GCTGTCGCCG TGGGCTGCCG ACCCGTATGA TGCCGCACGC 301 TTTGCCGGCG TGTTTTTCGC CGTTGTCGGA CTGACTTCCT GCGGCTTTGC 351 CGGTTTCAAC TTTTTGGGCA GACACCACGG GCGCAGCGTC GTCCTGATTC 401 TCATCGGCTG TATCGGGCTG ATTCCGACCG TACACTTTCT CAACCCCGCT 451 GCCGCCGCCT TTGCCGCCGC CGGACTGGTG CTGCACGGTT ATTCTTTGGC 501 TCGCCGGCGC GTGATTGCCG CCTCTTTTCT GCTCGGTACG GGTTGGACGC 551 TGATGTCGTT GGCAGCAGCT TATCCGGCGG CATTTGCCCT GATGCTGCCC 601 CTGCCCGTGC TGATGTTTTT CCGTCCGTGG CAAAGCAGGC GTTTGATGTT 651 GACGGCAGTC GCCTCGCTTG CCTTTGCCCT GCCGCTTATG ACCGTTTACC 701 CGCTGCTCTT GGCAAAAACG CAGCCCGCGC TGTTCGCGCA ATGGCTCGAC 751 GATCACGTTT TCGGTACGTT CGGCGGCGTG CGGCACATTC AGACGGCATT 801 CAGTTTGTTT TACTATCTGA AAAACCTGCT TTGGTTTGCA TTGCCTGCGC 851 TGCCGCTGGC GGTTTGGACG GTTTGCCGCA CGCGCCTGTT TTCGACCGAC 901 TGGGGGATTT TGGGCGTCGT CTGGATGCTT GCCGTTTTGG TGCTGCTTGC 951 CGTCAATCCG CAGCGTTTTC AGGATAACCT CGTCTGGCTG CTTCCGCCGC 1001 TTGCCCTGTT CGGCGCGGCG CAACTGGACA CCCTGAGACG CGGCGCGGCG 1051 GCGTTTGTCA ACTGGTTCGG CATTATGGCG TTCGGACTGT TTGCCGTGTT 1101 CCTGTGGACG GGCTTTTTCG CCATGAATTA CGGCTGGCCC GCCAAGCTTG 1151 CCGAACGCGC CGCCTATTTC AGCCCGTATT ATGTTCCTGA TATCGATCCC 1201 ATTCCGATGG CGGTTGCCGT ACTGTTCACA CCCTTGTGGC TGTGGGCGAT 1251 TACCCGCAAA AACATACGCG GCAGGCAGGC GGTTACCAAC TGGGCGGCAG 1301 GCGTTACCCT GACCTGGGCT TTGCTGATGA CGCTGTTCCT GCCGTGGCTG 1351 GACGCGGCGA AAAGCCACGC GCCCGTCGTC CGGAGTATGG AGGCATCGCT 1401 TTCCCCGGAA TTAAAACGGG AGCTTTCAGA CGGCATCGAG TGTATCGACA 1451 TAGGCGGCGG CGACCTACAC ACGCGGATTG TTTGGACGCA GTACGGCACA 1501 TTGCCGCACC GCGTCGGCGA TGTACAATGC CGCTACCGCA TCGTCCGCTT 1551 GCCCCAAAAC GCGGATGCGC CGCAAGGCTG GCAGACGGTC TGGCAGGGTG 1601 CGCGCCCGCG CAACAAAGAC AGTAAGTTCG CACTGATACG GAAAACCGGG 1651 GAAAATATAT TAAAAACAAC AGATTGA This encodes a protein having amino acid sequence <SEQ ID 598>: 1 MLTYTPPDAR PPAKTHEKPW LLLLMAFAWL WPGVFSHDLW NPDEPAVYTA 51 VEALAGSPTP LVAHLFGQID FGIPPVYLWV AAAFKHLLSP WAADPYDAAR 101 FAGVFFAWG LTSCGFAGFN FLGRHHGRSV VLILIGCIGL IPTVHFLNPA 151 AAAFAAAGLV LHGYSLARRR VIAASFLLGT GWTLMSLAAA YPAAFALMLP 201 LPVLMFFRPW QSRRLMLTAV ASLAFALPLM TVYPLLLAKT QPALFAQWLD 251 DHVFGTFGGV RHIQTAFSLF YYLKNLLWFA LPALPLAVWT VCRTRLFSTD 301 WGILGWWML AVLVLLAVNP QRFQDNLVWL LPPLALFGAA QLDSLRRGAA 351 AFVNWFGIMA FGLFAVFLWT GFFAMNYGWP AKLAERAAYF SPYYVPDIDP 401 IPMAVAVLFT PLWLWAITRK NIRGRQAVTN WAAGVTLTWA LLMTLFLPWL 451 DAAKSHAPW RSMEASLSPE LKRELSDGIE CIDIGGGDLH TRIVWTQYGT 501 LPHRVGDVQC RYRIVRLPQN ADAPQGWQTV WQGARPRNKD SKFALIRKTG 551 ENILKTTD* ORF141a and ORF141-1 show 98.2% identity in 553 aa overlap: oif141a . pep MLTYTPPDARPPAKTHEKPWLLLLMAFAWLWPGVFSHDLWNPDEPAVYTAVEALAGSPTP II I II I I I II II I I I I I II I I I I I II II II II I II I II II I II I I I I I I I II I I I II I I I orf141-1 MLTYTPPDARPPAKTHEKPWLLLLMAFAWLWPGVFSHDLWNPDEPAVYTAVEALAGSPTP orf141a . pep LVAHLFGQIDFGIPPVYLWVAAAFKHLLSPWAADPYDAARFAGVFFAWGLTSCGFAGFN II I II II I I II I I I I I II I I II I I I II II I II I I I II II I I II I I I : I I II I I I I II I orf141-1 LVAHLFGQTDFGIPPVYLWVAAAFKHLLSPWAADSYDAARFAGVFFAVIGLTSCGFAGFN orf141a. pep FLGRHHGRSWLILIGCIGLIPTVHFLNPAAAAFAAAGLVLHGYSLARRRVIAASFLLGT I I I II II I II II I I I I I I II I I :: I I I I I I I I I I I I I I I II I I I I I II I II I II II I I II orfl41-l FLGRHHGRSWLILIGCIGLIPVAHFLNPAAAAFAAAGLVLHGYSLARRRVIAASFLLGT orf14 la . pep GWTLMSLAAAYPAAFALMLPLPVLMFFRPWQSRRLMLTAVASLAFALPLMTVYPLLLAKT I I II I I I II I I I I I I I I I I I II I I II II II I II II II I I II I II I I II I I I II I I I I II I orf141-1 GWTLMSLAAAYPAAFALMLPLPVLMFFRPWQSRRLMLTAVASLAFALPLMTVYPLLLAKT orf141a . pep QPALFAQWLDDHVFGTFGGVRHIQTAFSLFYYLKNLLWFALPALPLAVWTVCRTRLFSTD I II I II I II I I II I I I II I II : I I I I I II I II I I I I I I II I II I I II I II I II II I I II orf141-1 QPALFAQWLDYHVFGTFGGVRHVQTAFSLFYYLKNLLWFALPALPLAVWTVCRTRLFSTD orf14la. pep WGILGWWMLAVLVLLAVNPQRFQDNLVWLLPPLALFGAAQLDSLRRGAAAFVNWFGIMA II I II I II I II I I II I II I I II I I I I I II I II I II II I I I I I I II I II I I I II I I I I II I orf141-1 WGILGWWMLAVLVLLAVNPQRFQDNLVWLLPPLALFGAAQLDSLRRGAAAFVNWFGIMA orf14 la. pep FGLFAVFLWTGFFAMNYGWPAKLAERAAYFSPYYVPDIDPIPMAVAVLFTPLWLWAITRK I I II I I I II I I I I I I I I I II I II I I II I I II I II II II II I II I II I I II II I I I II I I I orf141-1 FGLFAVFLWTGFFAMNYGWPAKLAERAAYFSPYYVPDIDPIPMAVAVLFTPLWLWAITRK orf141a . pep NIRGRQAVTNWAAGVTLTWALLMTLFLPWLDAAKSHAPWRSMEASLSPELKRELSDGIE II I I II II I I I I I I I I I II I II I I I I I I II I II I II I I I I I I I I II I I I II I I II II I II orfl41-l NIRGRQAVTNWAAGVTLTWALLMTLFLPWLDAAKSHAPWRSMEASLSPELKRELSDGIE orf141a . pep CIDIGGGDLHTRIVWTQYGTLPHRVGDVQCRYRIVRLPQNADAPQGWQTVWQGARPRNKD II I I I I I I I II I I I II I I I I I II I II II I II I II I I I II I I II I II I I I I II II I I II orf141-1 CIGIGGGDLHTRIVWTQYGTLPHRVGDVQCRYRIVLLPQNADAPQGWQTVWQGARPRNKD orf141a. pep SKFALIRKTGENI I I I I I II I I I II orfl41-l SKFALIRKIGENI Homology with a predicted ORF from Nsonorrhoeae ORF141 shows 95% identity over a 140aa overlap with a predicted ORF (ORF141ng) from N.gonorrhoeae: orfl41.pep DFGISPVYLWVAAAFKHLLSPWAADSYDVA 30 I II I II II II I II I II I II I II I MM orfl41ng WNPAEPAVYTAVEALAGSPTPLVAHLFGQTDFGIPPVYLWVAAAFKHLLSPWAAHPYDAA 126 orfl41.pep RFAGVFFAVIGLTSCGFAGFNFLGRHHGRXWLILIGCIGLIPVAHFLNPAAAAFAAAGL 90 I II II II I II II II I I I I II I II II I II I I II I I II I I I II I I I I : I I I I I I II II II orf14 lng RFAGVFFAVIGLTSCGFAGFNFLGRHHGRSWLIHIGCIGLIPVAHFFNPAAAAFAAAGL 186 orf141. pep VLHGYSLARRRVIAASFLLGTGWTLMSLAAAYPAAFALMLPLPVLMFFRP 140 I II II II I II II II II II I II I I I I I II I I I I II I II I II I I I II II I I I orf14lng VLHGYSLARRRVIAASFLLGTGWTLMSLAAAYPAAFALMLPLPVLMFFRPWQSRRLMLTA 246 An ORF141ng nucleotide sequence <SEQ ED 599> was predicted to encode a protein having amino acid sequence <SEQ ID 600>: 1 MPSEAVSARP LCEYLLHLAI RPFLLTLMLT YTPPDARPPA KTHEKPWLLL 51 LMAFAWLWPG VFSHDLWNPA EPAVYTAVEA LAGSPTPLVA HLFGQTDFGI 101 PPVYLWVAAA FKHLLSPWAA HPYDAARFAG VFFAVIGLTS CGFAGFNFLG 151 RHHGRSWLI HIGCIGLIPV AHFFNPAAAA FAAAGLVLHG YSLARRRVIA 201 ASFLLGTGWT LMSLAAAYPA AFALMLPLPV LMFFRPWQSR RLMLTAVASL 251 AFALPLMTVY PLLLAKTQPA LFAQWLNYHV FGTFGGVRHI QRAFSLFHYL 301 KNLLWFAPPG LPLAVWTVCR TRLFSTDWGI LGIVWMLAVL VLLAFNPQRF 351 QDNLVWLLPP LALFGAAQLD SLRRGAAAFV NWFGIMAFGL FAVFLWTGFF 401 AMNYGWPAKL AERAAYFSPY YVPDIDPIPM AVAVLFTPLW LWAITRKNIR 451 GRQAVTNWAA GVTLTWALLM TLFLPWLDAA KSHAP RSM EASFSPELKR 501 ELSDGIECIG IGGGDLHTRI VWTQYGTLPH RVGDVRCRYR IVRLPQNADA 551 PQGWQTVWQG ARPRNKDSKF ALIRKIGENI LKTTD* Further work revealed the following gonococcal DNA sequence <SEQ ID 601>: 1 ATGCTGACCT ATACCCCGCC CGATGCCCGC CCGCCCGCCA AAACCCACGA 51 AAAACCGTGG CTGCTGCTGT TGATGGCGTT TGCCTGGCTG TGGCCCGGCG 101 TGTTTTCCCA CGATTTGTGG AATCCTGCCG AACCTGCCGT CTATACCGCC 151 GTCGAAGCAC TGGCAGGCAG CCCCACCCCC TTGGTTGCCC ATCTGTTCGG 201 TCAAACCGAT TTCGGCATAC CGCCCGTGTA TCTTTGGGTT GCCGCCGCAT 251 TCAAACATTT GCTGTCGCCG TGGGCAGCCG ACCCGTATGA TGCCGCACGC 301 TTTGCAGGCG TATTTTTTGC CGTTATCGGA CTGACTTCTT GCGGCTTTGC 351 CGGTTTCAAC TTTTTGGGCA GACACCACGG GCGCAGCGTT GTTTTAATCC 401 ATATCGGCTG TATCGGGCTG ATTCCGGTTG CCCATTTCCT CAATCCcgcc 451 gccgccgcct tTGCCGCCGC CGGACTGGTG CTGCacggct actcgctgGC 501 ACGCCGGCGC GTGATtgccg cctctTtccT GCTCGGTACG GGTTGGACGT 551 TGATGTCGCT GGCGGCAGCT TATCCGGCGG CGTTTGCGCT GATGCTGCCC 601 CTGCCCGTGC TGATGTTTTT CCGTCCGTGG CAAAGCAGGC GTTTGATGTT 651 GACGGCAGTC GCCTCGCTTG CCTTTGCCCT GCCGCTTATG ACCGTTTACC 701 CGCTGCTCtt gGCAAAAACG CAGCCCGCGC TGTTTGCGCA ATGGCTCAAC 751 TATCACGTTT TCGGTACGTt cggcgGCGTG CGGCAcaTTC AGAggGCatT 801 Cagtttgttt cactatctgA AAaatctgct ttggttcgca ccgcccgggC 851 TGCCGCTGGC GGTTTGGACG GTTTGCCGCA CACGCCTGTT TTCGACCGAC 901 TGGGGGATTT TGGGCATTGT CTGGATGCTT GCCGTTTTGG TGCTGCTCGC 951 CTTTAATCCG CAGCGTTTTC AAGACAACCT CGTCTGGCTG CTGCCGCCGC 1001 TTGCCCTGTT CGGCGCGGCG CAACTGGACA GCCTGAGGCG CGGCGCGGCG 1051 GCTTTTGTCA ACTGGTTCGG CATTATGGCG TTCGGGCTGT TTGCCGTGTT 1101 CCTGTGGACG GGCTTTTTCG CCATGAATTA CGGCTGGCCC GCCAAGCTTG 1151 CCGAACGCGC CGCCTACTTC AGCCCGTATT ACGTTCCCGA CATCGATCCC 1201 ATTCCGATGG CGGTTGCCGT ACTGTTCACA CCCTTGTGGC TGTGGGCGAT 1251 TACCCGGAAA AACATACGCG GCAGGCAGGC GGTTACCAAC TGGGCGGCAG 1301 GCGTTACCCT GACCTGGGCT TTGCTGATGA CGCTGTTCCT GCCGTGGCTG 1351 GACGCGGCGA AAAGCCACGC GCCCGTCGTC CGGAGTATGG AGGCATCGTT 1401 TTCCCCGGAA TTAAAACGGG AGCTTTCAGA CGGCATCGAG TGTATCGGCA 1451 TAGGCGGCGG CGACCTGCAC ACGCGGATTG TTTGGACGCA GTACGGCACA 1501 TTGCCGCACC GCGTCGGCGA TGTCCGTTGC CGCTACCGTA TCGTCCGCCT 1551 GCCCCAAAAC GCGGATGCGC CGCAAGGCTG GCAGACGGTC TGGCAGGGTG 1601 CGCGCCCGCG CAACAAAGAC AGTAAGTTTG CACTGATACG GAAAATCGGG 1651 GAAAATATAT TAAAAACAAC AGATTGA This coπesponds to the amino acid sequence <SEQ ID 602; ORF141ng-l>: 1 MLTYTPPDAR PPAKTHEKPW LLLLMAFAWL WPGVFSHDLW NPAEPAVYTA 51 VEALAGSPTP LVAHLFGQTD FGIPPVYLWV AAAFKHLLSP WAADPYDAAR 101 FAGVFFAVIG LTSCGFAGFN FLGRHHGRSV VLIHIGCIGL IPVAHFLNPA 151 AAAFAAAGLV LHGYSLARRR VIAASFLLGT GWTLMSLAAA YPAAFALMLP 201 LPVLMFFRPW QSRRLMLTAV ASLAFALPLM TVYPLLLAKT QPALFAQWLN 251 YHVFGTFGGV RHIQRAFSLF HYLKNLLWFA PPGLPLAVWT VCRTRLFSTD 301 WGILGIVWML AVLVLLAFNP QRFQDNLVWL LPPLALFGAA QLDSLRRGAA 351 AFVNWFGIMA FGLFAVFLWT GFFAMNYGWP AKLAERAAYF SPYYVPDIDP 401 IPMAVAVLFT PLWLWAITRK NIRGRQAVTN WAAGVTLTWA LLMTLFLPWL 451 DAAKSHAPW RSMEASFSPE LKRELSDGIE CIGIGGGDLH TRIVWTQYGT 501 LPHRVGDVRC RYRIVRLPQN ADAPQGWQTV WQGARPRNKD SKFALIRKIG 551 ENILKTTD* ORF141ng-l and ORF141-1 show 97.5% identity in 553 aa overlap: orf141ng-l . pep MLTYTPPDARPPAKTHEKPWLLLLMAFAWLWPGVFSHDLWNPAEPAVYTAVEALAGSPTP I II II I II II II I I I II I II I II I I I I II II I II II I I I II I I I I I I II II I I I I I II I orfl41-l MLTYTPPDARPPAKTHEKPWLLLLMAFAWLWPGVFSHDLWNPDEPAVYTAVEALAGSPTP orf141ng-l . pep LVAHLFGQTDFGIPPVYLWVAAAFKHLLSPWAADPYDAARFAGVFFAVIGLTSCGFAGFN I I II II I II I I II I I I I I I I II II I I I I II II I I II I I I I I I II I I I I I I I I I I I II I I orf141-1 LVAHLFGQTDFGIPPVYLWVAAAFKHLLSPWAADSYDAARFAGVFFAVIGLTSCGFAGFN orf141ng-l .pep FLGRHHGRSWLIHIGCIGLIPVAHFLNPAAAAFAAAGLVLHGYSLARRRVIAASFLLGT II I I I I I I I I I I I I II I II I II I I I I I I II I I I I I II II I II I II II I II II II II I I I orf141-1 FLGRHHGRSWLILIGCIGLIPVAHFLNPAAAAFAAAGLVLHGYSLARRRVIAASFLLGT orfl41ng-l.pep GWTLMSLAAAYPAAFALMLPLPVLMFFRPWQSRRLMLTAVASLAFALPLMTVYPLLLAKT I I I I I I I I II II I I II I I II I II I II I I I I I I I II I I II I I I II I I I I II II I I II I I II orf141-1 GWTLMSLAAAYPAAFALMLPLPVLMFFRPWQSRRLMLTAVASLAFALPLMTVYPLLLAKT orfl41ng-l.pep QPALFAQWLNYHVFGTFGGVRHIQRAFSLFHYLKNLLWFAPPGLPLAVWTVCRTRLFSTD I II II I II I : I II I I I I I I I II : I I I II I M II II II II I : I I I II I II I II I I II I I orf141-1 QPALFAQWLDYHVFGTFGGVRHVQTAFSLFYYLKNLLWFALPALPLAVWTVCRTRLFSTD orf141ng-l . pep WGILGIVWMLAVLVLLAFNPQRFQDNLVWLLPPLALFGAAQLDSLRRGAAAFVNWFGIMA I I II I : I II I I I I I I II II I I I I I I II I I II I II I I II I I I I I I I I I II II I II I I I II orf141-1 WGILGWWMLAVLVLLAVNPQRFQDNLVWLLPPLALFGAAQLDSLRRGAAAFVNWFGIMA orfl41ng-l.pep FGLFAVFLWTGFFAMNYGWPAKLAERAAYFSPYYVPDIDPIPMAVAVLFTPLWLWAITRK II I I I II I I I I I I I I I I I I II I II I II I I I I I I I II II I I I II I II I I I II II I II II I I orf141-1 FGLFAVFLWTGFFAMNYGWPAKLAERAAYFSPYYVPDIDPIPMAVAVLFTPLWLWAITRK orf141ng-l .pep NIRGRQAVTNWAAGVTLTWALLMTLFLPWLDAAKSHAPWRSMEASFSPELKRELSDGIE I II II I II I II II II I I II I I II I II II II II I II II I I I I I II I I : I II II I I I II I II orf141-1 NIRGRQAVTNWAAGVTLTWALLMTLFLPWLDAAKSHAPWRSMEASLSPELKRELSDGIE orfl41ng-l.pep CIGIGGGDLHTRIVWTQYGTLPHRVGDVRCRYRIVRLPQNADAPQGWQTVWQGARPRNKD I II I II I II I I I I II II I II I I II II II : I I II I I I II II II I II I II I I II II II I I I orf141-1 CIGIGGGDLHTRIVWTQYGTLPHRVGDVQCRYRIVLLPQNADAPQGWQTVWQGARPRNKD orfl41ng-l.pep SKFALIRKIGENILKTTDX I II II I I II II I I orfl41-l SKFALIRKIGENIX Based on the presence of several putative transmembrane domains in the gonococcal protein, it is predicted that the proteins from Nmeningitidis and N.gonorrhoeae, and their epitopes, could be useful antigens for vaccines or diagnostics, or for raising antibodies. Example 72 The following partial DNA sequence was identified in Nmeningitidis <SEQ ID 603>: 1 .. CAATCCGCCA AATGGTTATC GGGCCAAACT CTAGTCGGCA CAGCAATTGG 51 GATACGCGGG CAGATAAAGC TTGGCGGCAA CCTGCATTAC GATATATTTA 101 CCGGCCGCGC ATTGAAAAAG CCCGAATTTT TCCAATCAAG GAAATGGGCA 151 AGCGGTTTTC AGGTAGGCTA TACGTTTTAA This coπesponds to the .amino acid sequence <SEQ ID 604; ORF142>: 1 .. QSA WLSGQT LVGTAIGIRG QIKLGGNLHY DIFTGRALKK PEFFQSRKWA 51 SGFQVGYTF* Further work revealed the complete nucleotide sequence <SEQ ED 605>: 1 ATGGATAATT CGGGTAGTGA GGCGACAGGA AAATACCAAG GAAATATCAC 51 TTTCTCTGCC GACAATCCTT TGGGACTGAG TGATATGTTC TATGTAAATT 101 ATGGACGTTC GATTGGCGGT ACGCCCGATG AGGAAAGTTT TGACGGCCAT 151 CGCAAAGAAG GCGGATCAAA CAATTACGCC GTACATTATT CAGCCCCTTT 201 CGGTAAATGG ACATGGGCAT TCAATCACAA TGGCTACCGT TACCATCAGG 251 CAGTTTCCGG ATTATCGGAA GTCTATGACT ATAATGGAAA AAGTTACAAT 301 ACTGATTTCG GCTTCAACCG CCTGTTGTAT CGTGATGCCA AACGCAAAAC 351 CTATCTCGGT GTAAAACTGT GGATGAGGGA AACAAAAAGT TACATTGATG 401 ATGCCGAACT GACTGTACAA CGGCGTAAAA CTGCGGGTTG GTTGGCAGAA 451 CTTTCCCACA AAGAATATAT CGGTCGCAGT ACGGCAGATT TTAAGTTGAA 501 ATATAAACGC GGCACCGGCA TGAAAGATGC TCTGCGCGCG CCTGAAGAAG 551 CCTTTGGCGA AGGCACGTCA CGTATGAAAA TTTGGACGGC ATCGGCTGAT 601 GTAAATACTC CTTTTCAAAT CGGTAAACAG CTATTTGCCT ATGACACATC 651 CGTTCATGCA CAATGGAACA AAACCCCGCT AACATCGCAA GACAAACTGG 701 CTATCGGCGG ACACCACACC GTACGTGGCT TCGACGGTGA AATGAGTTTG 751 TCTGCCGAGC GGGGATGGTA TTGGCGCAAC GATTTGAGCT GGCAATTTAA 801 ACCAGGCCAT CAGCTTTATC TTGGGGCTGA TGTAGGACAT GTTTCAGGAC 851 AATCCGCCAA ATGGTTATCG GGCCAAACTC TAGTCGGCAC AGCAATTGGG 901 ATACGCGGGC AGATAAAGCT TGGCGGCAAC CTGCATTACG ATATATTTAC 951 CGGCCGCGCA TTGAAAAAGC CCGAATTTTT CCAATCAAGG AAATGGGCAA 1001 GCGGTTTTCA GGTAGGCTAT ACGTTTTAA This coπesponds to the amino acid sequence <SEQ ID 606; ORF142-l>: 1 MDNSGSEATG KYQGNITFSA DNPLGLSDMF YVNYGRSIGG TPDEESFDGH 51 RKEGGSNNYA VHYSAPFGKW TWAFNHNGYR YHQAVSGLSE VYDYNGKSYN 101 TDFGFNRLLY RDAKRKTYLG VKLWMRETKS YIDDAELTVQ RRKTAGWLAE 151 LSHKEYIGRS TADFKLKYKR GTGMKDALRA PEEAFGEGTS RMKIWTASAD 201 VNTPFQIGKQ LFAYDTSVHA QWNKTPLTSQ DKLAIGGHHT VRGFDGEMSL 251 SAERGWYWRN DLSWQFKPGH QLYLGADVGH VSGQSAKWLS GQTLVGTAIG 301 IRGQIKLGGN LHYDIFTGRA LKKPEFFQSR KWASGFQVGY TF* Computer analysis of this amino acid sequence gave the following results: Homology with a predicted ORF from N.sonorrhoeae ORF142 shows 88.1% identity over a 59aa overlap with a predicted ORF (ORF142ng) from N.gonorrhoeae: orfl 42 . pep QSAKWLSGQTLVGTAIGIRGQIKLGGNLHY 30 I I I M I I I I I I : I I I I I I I I I I I I I I I I I I orf l42ng RGWYWRNDLSWQFKPGHQLYLGADVGHVSGQSAKWLSGQTLAGTAIGIRGQIKLGGNLHY 313 orf 142 . pep DIFTGRALKKPEFFQSRKWASGFQVGYTF 59 I I I I I I I I I I I I : I I : : I I : : I I I I I I : I orf l42ng DIFTGRALKKPEYFQTKKWVTGFQVGYSF 342 The complete length ORF142ng nucleotide sequence <SEQ ED 607> is: 1 ATGGATAATT CGGGTAGTGA GGCGACAGGA AAATACCAAG GAAATATCAC 51 TTTCTCTGCC GACAATCCTT TTGGACTGAG TGATATGTTC TATGTAAATT 101 ATGGACGTTC AATTGGCGGT ACGCCCGATG AGGAAAATTT TGACGGCCAT 151 CGCAAAGAAG GCGGATCAAA CAATTACGCC GTACATTATT CAGCCCCTTT 201 CGGTAAATGG ACATGGGCAT TCAATCACAA TGGCTACCGT TACCATCAGG 251 CGGTTTCCGG ATTATCGGAA GTCTATGACT ATAATGGAAA AAGTTACAAC 301 ACTGATTTCG GCTTCAACCG CCTGTTGTAT CGTGATGCCA AACGCAAAAC 351 CTATCTCAGT GTAAAACTGT GGACGAGGGA AACAAAAAGT TACATTGATG 401 ATGCCGAACT GACTGTACAA CGGCGTAAAA CCACAGGTTG GTTGGCAGAA 451 CTTTCCCACA AAGGATATAT CGGTCGCAGT ACGGCAGATT TTAAGTTGAA 501 ATATAAACAC GGCACCGGCA TGAAAGATGC TCTGCGCGCG CCTGAAGAAG 551 CCTTTGGCGA AGGCACGTCA CGTATGAAAA TTTGGACGGC ATCGGCTGAT 601 GTAAATACTC CTTTTCAAAT CGGTAAACAG CTATTTGCCT ATGACACATC 651 CGTTCATGCA CAATGGAACA AAACCCCGCT AACATCGCAA GACAAACTGG 701 CTATCGGCGG ACACCACACC GTACGTGGCT TCGACGGTGA AATGAGTTTG 751 CCTGCCGAGC GGGGATGGTA TTGGCGCAAC GATTTGAGCT GGCAATTTAA 801 ACCAGGCCAT CAGCTTTATC TTGGGGCTGA TGTAGGACAT GTTTCAGGAC 851 AATCCGCCAA ATGGTTATCG GGCCAAACTC TAGCCGGCAC AGCAATTGGG 901 ATACGCGGGC AGATAAAGCT TGGCGGCAAC CTGCATTACG ATATATTTAC 951 CGGCCGTGCA TTGAAAAAGC CCGAATATTT TCAGACGAAG AAATGGGTAA 1001 CGGGGTTTCA GGTGGGTTAT TCGTTTTGA This encodes a protein having amino acid sequence <SEQ ID 608>: 1 MDNSGSEATG KYQGNITFSA DNPFGLSDMF YVNYGRSIGG TPDEENFDGH 51 RKEGGSNNYA VHYSAPFGKW TWAFNHNGYR YHQAVSGLSE VYDYNGKSYN 101 TDFGFNRLLY RDAKRKTYLS VKLWTRETKS YIDDAELTVQ RRKTTGWLAE 151 LSHKGYIGRS TADFKLKYKH GTGMKDALRA PEEAFGEGTS RMKIWTASAD 201 VNTPFQIGKQ LFAYDTSVHA QWNKTPLTSQ DKLAIGGHHT VRGFDGEMSL 251 PAERGWYWRN DLSWQFKPGH QLYLGADVGH VSGQSAKWLS GQTLAGTAIG 301 IRGQIKLGGN LHYDIFTGRA LKKPEYFQTK KWVTGFQVGY SF* The underlined sequence (aromatic-Xaa-aromatic amino acid motif) is usually found at the C-terminal end of outer membrane proteins. ORF142ng and ORF142-1 show 95.6% identity over 342aa overlap: orf142-1. pep MDNSGSEATGKYQGNITFSADNPLGLSDMFYVNYGRSIGGTPDEESFDGHRKEGGSNNYA I II I I II I I I II I I I II I II I II : II I I I I I II I I I I I II I I I II : I I I II I I II I II II orfl42ng-l MDNSGSEATGKYQGNITFSADNPFGLSDMFYVNYGRSIGGTPDEENFDGHRKEGGSNNYA orf142-1. pep VHYSAPFGKWTWAFNHNGYRYHQAVSGLSEVYDYNGKSYNTDFGFNRLLYRDAKRKTYLG I I I I I II I II I I II II II II I I II I II II II I I II I II I I I I II I I II I I I II I I I I I I : orfl42ng-l VHYSAPFGKWTWAFNHNGYRYHQAVSGLSEVYDYNGKSYNTDFGFNRLLYRDAKRKTYLS orf142-1. pep VKLWMRETKSYIDDAELTVQRRKTAGWLAELSHKEYIGRSTADFKLKYKRGTGMKDALRA I I I I I I II I I I I II I I II I II I I : I II II I II I I I I I I I II II II I I : II II II II II orfl42ng-l VKLWTRETKSYIDDAELTVQRRKTTGWLAELSHKGYIGRSTADFKLKYKHGTGMKDALRA orf142-1. pep PEEAFGEGTSRMKIWTASADVNTPFQIGKQLFAYDTSVHAQWNKTPLTSQDKLAIGGHHT I I I I I I I II I I I I II II II I II I II I I II II I II I II I I I I I I I II II I I I I I II II II I orfl42ng-l PEEAFGEGTSRMKIWTASADVNTPFQIGKQLFAYDTSVHAQWNKTPLTSQDKLAIGGHHT orf142-1. pep VRGFDGEMSLSAERGWYWRNDLSWQFKPGHQLYLGADVGHVSGQSAKWLSGQTLVGTAIG II I II II II I I I I II II II I I II I II I II II I I II II I II I I I I II I I I II II : I I II I orfl42ng-l VRGFDGEMSLPAERGWYWRNDLSWQFKPGHQLYLGADVGHVSGQSAKWLSGQTLAGTAIG orf142-1. pep IRGQIKLGGNLHYDIFTGRALKKPEFFQSRKWASGFQVGYTF II I I I II I I I II II I I I I I I I I II I : II : : I I : : I I I I II : I orfl42ng-l IRGQIKLGGNLHYDIFTGRALKKPEYFQTKKWVTGFQVGYSF In addition, ORF142ng is homologous to the HecB protein of E.chrysanthemi: gi 11772622 (L39897) HecB [Er inia chrysanthemi] Length = 558 Score = 119 bits (295), Expect = 3e-26 Identities = 88/346 (25%), Positives = 151/346 (43%), Gaps = 22/346 (6%) Query: 2 DNSGSEATGKYQGNITFSADNPFGLSDMFYVNYGRΞIGGTPDEENFDGHRKEGGSNNYAV 61 DNSG ++TG+ Q N + + DN FGL+D ++++ G S + + D + G Sbjct: 230 DNSGQKSTGEEQLNGSLALDNVFGLADQWFISAGHS SRFATSHDAESLQAG 280 Query: 62 HYSAPFGKWTWAFNHNGYRYHQAVSGLSEVYDYNGKSYNTDFGFNRLLYRDAKRKTYLSV 121 +S P+G W +N++ RY + G S F +R+++RD KT ++ Sbjct: 281 -FSMPYGYWNLGYNYSQSRYRNTFINRDFPWHSTGDSDTHRFSLSRWFRDGTMKTAIAG 339 Query: 122 KLWTRETKSYIDDAELTVQRRKTTGWLAELSHKGYIGRSTADFKLKYKHGTGMKDALRAP 181 R +Y++ + L RK + ++H + A F Y G + Sbjct: 340 TFSQRTGNNYLNGSLLPSSSRKLSSVSLGVNHSQKLWGGLATFNPTYNRGVRWLGSETDT 399 Query: 182 EEAFGEGTSRMKIWTASADVNTPFQIGKQLFAYDTSVHAQWNKTPLTSQDKLAIGGHHTV 241 +++ E + WT SA P Y S++ Q++ L ++L +GG ++ Sbjct: 400 DKSADEPRAEFNKWTLSASYYHPV TDSITYLGSLYGQYSARALYGSEQLTLGGESSI 456 Query: 242 RGFDGEMSLPAERGWYWRNDLSWQFKP GHQLYLGA-DVGHVSGQSAKWLSGQTLAG 296 RGF E RG YWRN+L+WQ G+ ++ A D GH+ + +L G Sbjct: 457 RGF-REQYTSGNRGAYWRNELNWQAWQLPVLGNVTFMAAVDGGHLYNHKQDNSTAASLWG 515 Query : 297 TAIGIRGQIKLGGNLHYDIFTGRALKKPEYFQTKKWVTGFQVGYSF 342 A+G+ + + G + P + Q V G++VG SF Sbjct: 516 GAVGMTVASRW LSQQVTVGWPISYPAWLQPDTMWGYRVGLSF 558 On the basis of this analysis, it is predicted that the proteins from N. meningitidis .and N.gonorrhoeae, .and their epitopes, could be useful antigens for vaccines or diagnostics, or for raising antibodies. Example 73 The following partial DNA sequence was identified in N. meningitidis <SEQ ID 609>: 1 ATGCGGACGA AATGGTCAGC AGTGAGAAGC TGCTJACTTG GgCGGACACC 51 GCCGACATCG ATACCGCTTT GAACCTGTTG TACCGTTTGC AAAAACTCGA 101 ATTCCTCTAT GGCGATGAAA ACGGTCATTC AGACGGCATC AATTTGwCGG 151 ACGAGCAATT GCCGTTGCTG ATGGAACAAT TGTCCGGCAG CGGTAAGGCG 201 TTATTGGTCG ATCGGAACGG TCTGTATCTT GCCAACGCCA ATTTCCATCA 251 TGAGGCGGCG GAAGAGTTGG GGTTGTTGGC GGCAGAAGTC GCACAGATGG 301 AAAAGAAATA CCGGCTGCTG ATTAAGAACA AC. This coπesponds to the amino acid sequence <SEQ ID 610; ORF143>: 1 MRTKWSAVRS C7WADTADID TALNLLYRLQ KLEFLYGDEN GHSDGINLXD 51 EQLPLLMEQL SGSGKALLVD RNGLYLANAN FHHEAAEELG LLAAEVAQME 101 KKYRLLIKNN .. Further work revealed the complete nucleotide sequence <SEQ ID 611>: 1 ATGGAATCAA CACTTTCACT ACAAGCAAAT TTATATCCCC GCCTGACTCC 51 TGCCGGTGCA TTTTATGCCG TATCCAGCGA TGCCCCCAGT GCCGGTAAAA 101 CTTTGTTGCA CAGCCTGTTG AAAGCAGATG CGGACGAAAT GGTCAGCAGT 151 GAGAAGCTGC TTACTTGGGC GGACACCGCC GACATCGATA CCGCTTTGAA 201 CCTGTTGTAC CGTTTGCAAA AACTCGAATT CCTCTATGGC GATGAAAACG 251 GTCATTCAGA CGGCATCAAT TTGTCGGACG AGCAATTGCC GTTGCTGATG 301 GAACAATTGT CCGGCAGCGG TAAGGCGTTA TTGGTCGATC GGAACGGTCT 351 GTATCTTGCC AACGCCAATT TCCATCATGA GGCGGCGGAA GAGTTGGGGT 401 TGTTGGCGGC AGAAGTCGCA CAGATGGAAA AGAAATACCG GCTGCTGATT 451 AAGAACAACC TGTATATCAA CAATAACGCT TGGGGCGTTT GCGATCCTTC 501 CGGTCAGAGC GAATTGACAT TTTTCCCATT GTATATCGGT TCAACCAAAT 551 TTATTTTGGT TATCGGCGGC ATTCCCGATT TGGGCAAAGA GGCATTTGTT 601 ACTTTGGTAA GGATTTTATA CCGCCGTTAC AGCAACCGCG TGTAA This coπesponds to the amino acid sequence <SEQ ID 612; ORF143-1 > • 1 MESTLSLQAN LYPRLTPAGA FYAVSSDAPS AGKTLLHSLL KADADEMVSS 51 EKLLΓWADTA DIDTALNLLY RLQKLEFLYG DENGHSDGIN LSDEQLPLLM 101 EQLSGSGKAL LVDRNGLYLA NANFHHEAAE ELGLLAAEVA QMEKKYRLLI 151 KNNLYINNNA WGVCDPSGQS ELTFFPLYIG STKFILVIGG IPDLGKEAFV 201 TLVRILYRRY SNRV* Computer analysis of this amino acid sequence gave the following results: Homology with a predicted ORF from N. meningitidis (sfrain A) ORF143 shows 92.4% identity over a 105aa overlap with an ORF (ORF143a) from strain A of N. meningitidis: 10 20 30 orr143. pep MRTKWSAVRSCTWADTADIDTALΝLLYRLQKLEFL M : III I II I I I I I I I I I II I I II I I orf143a GAFYAVSSDXPSAGKTLLHSLLKADADEMVSSEKLLTWAXTADIDTALΝLLYRLQKLEFL 20 30 40 50 60 70 40 50 60 70 80 90 orf143. pep YGDEΝGHSDGIΝLXDEQLPLLMEQLSGSGKALLVDRΝGLYLAΝAΝFHHEAAEELGLLAAE I II II II I II I II II I I II I II I I I II II I I I I I I I II I I I I I I I I I I I I II I M I I I I orf 143a YGDENGHSDGINLSDEQLPLLMEQLSGSGKALLVDRNGLYLANANFHHEAAEELGLLAAE 80 90 100 110 120 130 100 110 orf143.pep VAQMEKKYRLLIKNN I I I I I I II I I I I I I orfl43a VAQMEKKYRLXIKNNLYINNNAWGVCDPSGQSELTFFPLYIGSTKFILVIGGIPDLGKEA 140 150 160 170 180 190 The complete length ORF 143 a nucleotide sequence <SEQ ID 613> is: 1 ATGGAATCAA CANTTTCACT ACAAGCAAAT TTATATCNCC GCCTGACTCC 51 TGCCGGTGCA TTTTATGCCG TATCCAGCGA TGNCCCCAGT GCCGGTAAAA 101 CTTTGTTGCA CAGCCTGTTG AAAGCGGATG CGGACGAAAT GGTNAGCAGT 151 GAGAAGCTGC TTACCTGGGC GGANACCGCC GACATCGATA CCGCTTTGAA 201 CCTGTTGTAC CGTTTGCAAA AACTCGAATT CCTCTATGGC GATGAAAACG 251 GTCATTCAGA CGGCATCAAT TTGTCGGACG AGCAATTGCC GTTGCTGATG 301 GAACAATTGT CCGGCAGCGG TAAGGCGTTA TTGGTCGATC GGAACGGTCT 351 GTATCTTGCC AACGCCAATT TCCATCATGA GGCGGCGGAA GAGTTGGGGT 401 TGTTGGCGGC AGAAGTCGCA CAGATGGAAA AGAAATACCG GCTGCNNATT 451 AAGAACAACC TGTATATCAA CAATAACGCT TGGGGCGTTT GCGATCCTTC 501 CGGTCAGAGC GAATTGACAT TTTTCCCATT GTATATCGGT TCAACCAAAT 551 TTATTTTGGT TATCGGCGGC ATTCCCGATT TGGGCAAAGA GGCATTTGTT 601 ACTTTGGTAA GGATNTTATA CCNCCNGTTA CAGCAACCGC GTGTAAAACT 651 TGGGAGAGAG GANGGGTTAT GCAGCAATTA TTGA This encodes a protein having amino acid sequence <SEQ ID 614>: 1 MESTXSLQAN LYXRLTPAGA FYAVSSDXPS AGKTLLHSLL KADADEMVSS 51 EKLLTWAXTA DIDTALNLLY RLQKLEFLYG DENGHSDGIN LSDEQLPLLM 101 EQLSGSGKAL LVDRNGLYLA NANFHHEAAE ELGLLAAEVA QMEKKYRLXI 151 KNNLYINNNA WGVCDPSGQS ELTFFPLYIG STKFILVIGG IPDLGKEAFV 201 TLVRXLYXXL QQPRVKLGRE XGLCSNY* ORF143a and ORF143-1 show 97.1% identity in 207 aa overlap: orf 143a . pep MESTXSLQANLYXRLTPAGAFYAVSSDXPSAGKTLLHSLLKADADEMVΞSEKLLTWAXTA I I I I I I I II I I II I I I I I II II I II II I I II I I II II II II I I I I I II II I I I I II orf143-1 MESTLSLQANLYPRLTPAGAFYAVSSDAPSAGKTLLHSLLKADADEMVSSEKLLTWADTA orf143a. pep DIDTALNLLYRLQKLEFLYGDENGHSDGINLSDEQLPLLMEQLSGSGKALLVDRNGLYLA I I I II I I I II II I II I I I I I II II I I I I II I I I I I I I I I I II I I II I I I II I II I II I I I orf143-1 DIDTALNLLYRLQKLEFLYGDENGHSDGINLSDEQLPLLMEQLSGSGKALLVDRNGLYLA orf143a. pep NANFHHEAAEELGLLAAEVAQMEKKYRLXIKNNLYINNNAWGVCDPSGQSELTFFPLYIG II I I II I II II I I I I I I II I I I I I II I I I I II I II I I I II I I I I II I I II II I I I I II I orf143-1 NANFHHEAAEELGLLAAEVAQMEKKYRLLIKNNLYINNNAWGVCDPSGQSELTFFPLYIG orf143a. pep STKFILVIGGIPDLGKEAFVTLVRXLY I II I I I II I II I II II I II I I I I I II orf 143-1 STKFILVIGGIPDLGKEAFVTLVRILY Homology with a predicted ORF from N.sonorrhoeae ORF143 shows 95.5% identity over a l lOaa overlap with a predicted ORF (ORF143ng) from N.gonorrhoeae: orf143.pep MRTKWSAVRSCTWADTADIDTALNLLYRLQKLEFLYGDENGHSDGINLXDEQLPLLMEQL 60 I I I II I I I I I I : I II II I II I I II I I II I II I I II I I I I II II II II I I I II I II II I orf143ng MRTKWSAVRSCSRADTADIDTALNLLYRLQKLEFLYGDENGHSDGINLSDEQLPLLMEQL 60 orf143.pep SGSGKALLVDRNGLYLANANFHHEAAEELGLLAAEVAQMEKKYRLLIKNN 110 II I I II II I I I II I I II I I I I I I I: II I II I I II I I I I I I I II I I II: I I orf143ng SGSGKALLVDRNGLYLANANFHHESAEELGLLAAEVAQMEKKYRLLIRNNLYINNNAWGV 120 An ORF143ng nucleotide sequence <SEQ ED 615> was predicted to encode a protein having amino acid sequence <SEQ LD 616>: 1 MRTKWSAVRS CSRADTADID TALNLLYRLQ KLEFLYGDEN GHSDGINLSD 51 EQLPLLMEQL SGSGKALLVD RNGLYLA AN FHHESAEELG LLAAEVAQME 101 KKYRLLIRNN LYINNNAWGV CDPSGQSELT FFPLYIGSTK FILVIAGIPD 151 LSKGGICYFG KDFIPPLQQP RVKLGTGGIM RQLLISILED LNNTSTDIIA 201 SAVISTDGLP MATMLPSHLN SDRVGAISAT LLALGSRSVQ ELACGELEQV 251 MIKGKSGYIL LSQAGKDAVL VLVAKETGRL GLILLDAKRA ARHIAEAI* Further work revealed the following gonococcal DNA sequence <SEQ ID 617>: 1 ATGGAATCAA CACTTTCACT ACAAGCGAAT TTATATCCCT GCCTGACTCC 51 TGCCGGTGCA TTTTATGCCG TATCCAGCGA TGCCCCCAGT GCCGGTAAAA 101 CTTTGTTGCG CAGCCTGTTG AAAGCGGATG CGGACGAAGT GGTCAGCAGT 151 GAGAAGCTGC TCGCGGCGGA CACCGCCGAC ATCGATACCG CTTTGAACCT 201 GTTGTACCGT TTGCAAAAAC TCGAATTCCT CTATGGCGAT GAAAACGGTC 251 ATTCAGACGG CATCAATTTG TCGGACGAGC AATTGCCGTT GCTGATGGAA 301 CAATTGTCCG GCAGCGGTAA GGCATTATTG GTCGATCGGA ACGGTCTGTA 351 TCTTGCCAAC GCCAATTTCC ATCATGAGTC GGCGGAAGAG TTGGGGTTGT 401 TGGCGGCAGA AGTCGCACAG ATGGAAAAGA AATACCGGCT GCTGATTAGG 451 AACAACCTGT ATATCAACAA TAACGCTTGG GGCGTTTGCG ATCCTTCCGG 501 TCAGAGCGAA TTGACATTTT TCCCATTGTA TATCGGTTCA ACCAAATTTA 551 TTTTGGTTAT CGCCGGCATT CCCGATTTGA GCAAAGAGGC ATTTGTTACT 601 TTGGTAAGGA TTTTATACCG CCGTTACAGC AACCGCGTGT AA This coπesponds to the amino acid sequence <SEQ ID 618; ORF143ng-l>: 1 MESTLSLQAN LYPCLTPAGA FYAVSSDAPS AGKTLLRSLL KADADEWSS 51 EKLLAADTAD IDTALNLLYR LQKLEFLYGD ENGHSDGINL SDEQLPLLME 101 QLSGSGKALL VDRNGLYLAN ANFHHESAEE LGLLAAEVAQ MEKKYRLLIR 151 NNLYINNNAW GVCDPSGQSE LTFFPLYIGS TKFILVIAGI PDLSKEAFVT 201 LVRILYRRYS NRV* ORF143ng-l and ORF143-1 show 95.8% identity in 214 aa overlap: orfl43ng-l.pep MESTLSLQANLYPCLTPAGAFYAVSSDAPSAGKTLLRSLLKADADEWSSEKLLA-ADTA 59 I II II I I II I II I I I II II I I II I II II I I I I II I : II II II I II : II II I II: II II orf143-1 MESTLSLQANLYPRLTPAGAFYAVSSDAPSAGKTLLHSLLKADADEMVSSEKLLTWADTA 60 orfl43ng-l . pep DIDTALNLLYRLQKLEFLYGDENGHSDGINLSDEQLPLLMEQLSGSGKALLVDRNGLYLA 119 I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I orf 143-1 DIDTALNLLYRLQKLEFLYGDENGHSDGINLSDEQLPLLMEQLSGSGKALLVDRNGLYLA 120 orf 143ng-l . pep NANFHHESAEELGLLAAEVAQMEKKYRLLIRNNLYINNNAWGVCDPSGQSELTFFPLYIG 17 9 I I I I I I I : I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I orf 143-1 NANFHHEAAEELGLLAAEVAQMEKKYRLLIKNNLYINNNAWGVCDPSGQSELTFFPLYIG 180 orf l43ng-l . pep STKFILVIAGIPDLSKEAFVTLVRILYRRYSNRV 213 I I I I I I I M I I I I M I I I I I I I I I I I I I I I I I I I orf 143-1 STKFILVIGGIPDLGKEAFVTLVRILYRRYSNRV 214 Based on the presence of the putative transmembrane domains in the gonococcal protein, it is predicted that the proteins from N. meningitidis .and N.gonorrhoeae, and their epitopes, could be useful antigens for vaccines or diagnostics, or for raising antibodies. Example 74 The following partial DNA sequence was identified in N. meningitidis <SEQ ID 619>: 1 ATGACCTTTT TACAACGTTT GCAAGGTTTG GCAGACAATA AAATCTGTGC 51 GTTTGCATGG TTCGTCGTCC GCCGCTTTGA TGAAGAACGC GTACCGCAGr 101 CGGCGGCAAG CATGACGTTT ACGACGCTGC TGGCACTCGT CCCCGTGCTG 151 ACCGTGATGG TGGCGGTCGC TTCGATTTTC CCCGTGTTCG ACCGCTGGTC 201 GGATTCGTTC GTCTCCTTCG TCAACCAAAC CATTGTGCCG CA.GGCGCGG 251 ACATGGTGTT CGACTATATC AATGCGTTCC GCGAGCAGGC GAACCGGCTG 301 ACGGCAATCG GCAGCGTGAT GCTGGTCGTT ACCTCGCTGA TGCTGATTCG 351 GACGATAGAC AATACGTTCA ACCGCATCTG GaCGGGTCAA TyCCAGCGT 401 CCGTGGATG . This coπesponds to the amino acid sequence <SEQ ID 620; ORF144>: 1 MTFLQRLQGL ADNKICAFAW FWRRFDEER VPQXAASMTF TTLLALVPVL 51 TVMVAVASIF PVFDRWSDSF VSFVNQTIVP XGADMVFDYI NAFREQANRL 101 TAIGSVMLW TSLMLIRTID NTFNRIWRVX XQRPWM... Further work revealed the complete nucleotide sequence <SEQ ID 621 > • 1 ATGACCTTTT TACAACGTTT GCAAGGTTTG GCAGACAATA AAATCTGTGC 51 GTTTGCATGG TTCGTCGTCC GCCGCTTTGA TGAAGAACGC GTACCGCAGG 101 CGGCGGCAAG CATGACGTTT ACGACGCTGC TGGCACTCGT CCCCGTGCTG 151 ACCGTGATGG TGGCGGTCGC TTCGATTTTC CCCGTGTTCG ACCGCTGGTC 201 GGATTCGTTC GTCTCCTTCG TCAACCAAAC CATTGTGCCG CAGGGCGCGG 251 ACATGGTGTT CGACTATATC AATGCGTTCC GCGAGCAGGC GAACCGGCTG 301 ACGGCAATCG GCAGCGTGAT GCTGGTCGTT ACCTCGCTGA TGCTGATTCG 351 GACGATAGAC AATACGTTCA ACCGCATCTG GCGGGTCAAT TCCCAGCGTC 401 CGTGGATGAT GCAGTTTCTC GTCTATTGGG CTTTACTGAC GTTCGGGCCG 451 CTGTCTTTGG GCGTGGGCAT TTCCTTTATG GTCGGCTCGG TACAGGATGC 501 CGCGCTTGCC TCAGGTGCGC CGCAGTGGTC GGGCGCGTTG CGAACGGCGG 551 CGACGCTGAC CTTCATGACG CTTTTGCTGT GGGGGCTGTA CCGCTTCGTG 601 CCAAACCGCT TCGTTCCCGC GCGGCAGGCG TTTGTCGGGG CTTTGGCAAC 651 AGCGTTTTGT CTGGAAACCG CGCGCTCCCT CTTCACTTGG TATATGGGCA 701 ATTTCGACGG CTACCGCTCG ATTTACGGCG CGTTTGCCGC CGTGCCGTTT 751 TTTCTGTTGT GGCTGAACCT GTTGTGGACG CTGGTCTTGG GCGGCGCGGT 801 GCTGACTTCT TCACTCTCCT ACTGGCAGGG AGAAGCGTTC CGCAGGGGCT 851 TCGACTCGCG CGGACGGTTT GACGACGTGT TGAAAATCCT GCTGCTTCTG 901 GATGCGGCGC AAAAAGAAGG CAAAGCCTTG CCTGTTCAGG AGTTCAGACG 951 GCATATCAAT ATGGGCTACG ACGAGTTGGG CGAGCTTTTG GAAAAGCTGG 001 CGCGGCACGG CTACATCTAT TCCGGCAGAC AGGGTTGGGT GTTGAAAACG 051 GGGGCGGATT CGATTGAGTT GAACGAACTC TTCAAGCTCT TCGTTTACCG 101 TCCGTTGCCT GTGGAAAGGG ATCATGTGAA CCAAGCTGTC GATGCGGTAA 151 TGACACCGTG TTTGCAGACT TTGAACATGA CGCTGGCAGA GTTTGACGCT 201 CAGGCGAAAA AACGGCAGTA G This coπesponds to the amino acid sequence <SEQ ID 622; ORF144-l>: 1 MTFLQRLQGL ADNKICAFAW FWRRFDEER VPQAAASMTF TTLLALVPVL 51 TVMVAVASIF PVFDRWSDSF VSFVNQTIVP QGADMVFDYI NAFREQANRL 101 TAIGSVMLW TSLMLIRTID NTFNRIWRVN SQRPWMMQFL VYWALLTFGP 151 LSLGVGISFM VGSVQDAALA SGAPQWSGAL RTAATLTFMT LLLWGLYRFV 201 PNRFVPARQA FVGALATAFC LETARSLFTW YMGNFDGYRS IYGAFAAVPF 251 FLLWLNLLWT LVLGGAVLTS SLSYWQGEAF RRGFDSRGRF DDVLKILLLL 301 DAAQKEGKAL PVQEFRRHIN MGYDELGELL EKLARHGYIY SGRQGWVLKT 351 GADSIELNEL FKLFVYRPLP VERDHVNQAV DAVMTPCLQT LNMTLAEFDA 401 QAKKRQ* Computer analysis of this amino acid sequence gave the following results: Homology with a predicted ORF from N.meninsitidis (strain Al ORF144 shows 96.3% identity over a 136aa overlap with an ORF (ORF144a) from strain A of N. meningitidis: 10 20 30 40 50 60 orf 144 . pep MTFLQRLQGLADΝKICAFAWFWRRFDEERVPQXAASMTFTTLLALVPVLTVMVAVASIF II II I I I I II I I II I II I I I I II I II II II I II I I I I I I I II II II I II I I II I I I II I orf144a MTFLQRLQGLADΝKICAFAWFWRRFDEERVPQAAASMTFTTLLALVPVLTVMVAVASIF 10 20 30 40 50 60 70 80 90 100 110 120 orf144.pep PVFDRWSDSFVSFVΝQTIVPXGADMVFDYIΝAFREQAΝRLTAIGSVMLWTSLMLIRTID I I II I II I I I II I I II II I I I I II I I II I I I II I I II I II I I I II II I I I I I I II II I orfl44a PVFDRWSDSFVSFVΝQTIVPQGADMVFDYIΝAFREQAΝRLTAIGSVMLWTSXMLIRTID 70 80 90 100 110 120 130 orf144. pep ΝTFΝRIWRVXXQRPWM I I I I II II I II II I orf144a ΝTFΝRIWRVΝSQRPWMMQFLVYWALLTFGPLSLGVGISFXVGSVQDAALASGAPQWSGAL 130 140 150 160 170 180 The complete length ORF144a nucleotide sequence <SEQ ID 623> is: 1 ATGACCTTTT TACAACGTTT GCAAGGTTTG GCAGACAATA AAATCTGTGC 51 GTTTGCATGG TTCGTCGTCC GCCGCTTTGA TGAAGAACGC GTACCGCAGG 101 CGGCGGCAAG CATGACGTTT ACGACACTGC TGGCACTCGT CCCCGTGCTG 151 ACCGTGATGG TGGCGGTCGC TTCGATTTTC CCCGTGTTCG ACCGNTGGTC 201 GGATTCGTTC GTCTCCTTCG TCAACCAAAC CATTGTGCCG CAGGGCGCGG 251 ACATGGTNTT CGACTATATC AATGCGTTCC GCGAGCAGGC GAACCGGCTG 301 ACGGCAATCG GCAGCGTGAT GCTGGTCGTT ACCTCGCNGA TGCTGATTCG 351 GACGATAGAC AATACGTTCA ACCGCATCTG GCGGGTCAAT TCCCAGCGTC 401 CGTGGATGAT GCAGTTTCTC GTCTATTGGG CTTTACTGAC GTTCGGGCCG 451 CTGTCTTTGG GCGTGGGCAT TTCCTTTATN GTCGGCTCGG TACAGGATGC 501 CGCGCTTGCC TCAGGTGCGC CGCAGTGGTC GGGCGCGTTG CGAACGGCGG 551 CGACGCTGAN CTTCATGACG CTTTTGCTGT GGGGGCTGTA CCGCTNCGTG 601 CCAAACCGCT TCGTTCCCGC GCGGCANGCG TTTGTCGGGG CTTTGGCAAC 651 AGCGTTCTGT CTGGAAACCG CGCGTTCCCT CTTTACTTGG TATATGGGCA 701 ATTTCGACGG CTACCGCTCG ATTTACGGNG CGTTTGCCGC CGTGCCGTTT 751 TTTCTGTTGT GGCTGAACCT GTTGTGGACG CTGGTCTTGG GCGGCGCGGT 801 GCTGACTTCT TCACTCTCCT ACTGGCAGGG AGAAGCGTTC CGCAGGGNCT 851 TCGACTCGCG CGGACGGTTT GACGACGTGT TGAAAATCCT GCTGCTTCTG 901 GATGCGGCGC AAAAAGAAGG CNAAGCCTTG CCTGTTCAGG AGTTCAGACG 951 GCATATCAAT ATGGGCTACG ACGAGTTGGG CGAGCTTTTG GAAAAGCTGG 1001 CGCGGCACGG CTACATCTAT TCCGGCAGAC AGGGTTGGGT GTTGAAAACG 1051 GGGGCGGATT CGATTGAGTT GAACGAACTC TTCAAGCTCT TCGTTTACCG 1101 TCCGTTGCCT GTGGAAAGGG ATCATGTGAA CCAAGCTGTC GATGCGGTAA 1151 TGATGCCGTG TTTGCAGACT TTGAACATGA CGCTGGCAGA GTTTGACGCT 1201 CAGGCGAAAA AACAGCAGCA ATCTTGA This encodes a protein having amino acid sequence <SEQ ID 624>: 1 MTFLQRLQGL ADNKICAFAW FWRRFDEER VPQAAASMTF TTLLALVPVL 51 TVMVAVASIF PVFDRWSDSF VSFVNQTIVP QGADMVFDYI NAFREQANRL 101 TAIGSVMLW TSXMLIRTID NTFNRIWRVN SQRPWMMQFL VYWALLTFGP 151 LSLGVGISFX VGSVQDAALA SGAPQWSGAL RTAATLXFMT LLLWGLYRXV 201 PNRFVPARXA FVGALATAFC LETARSLFTW YMGNFDGYRS IYGAFAAVPF 251 FLLWLNLLWT LVLGGAVLTS SLSYWQGEAF RRXFDSRGRF DDVLKILLLL 301 DAAQKEGXAL PVQEFRRHIN MGYDELGELL EKLARHGYIY SGRQGWVLKT 351 GADSIELNEL FKLFVYRPLP VERDHVNQAV DAVMMPCLQT LNMTLAEFDA 401 QAKKQQQS* ORF144a and ORF144-1 show 97.8% identity in 406 aa overlap: or 144a. pep MTFLQRLQGLADNKICAFAWFWRRFDEERVPQAAASMTFTTLLALVPVLTVMVAVASIF I I II I I II II II I I I I I I II I II I II I I I I II II II I I I I I II I I I I I II II I I I I I I I I orf144-1 MTFLQRLQGLADNKICAFAWFWRRFDEERVPQAAASMTFTTLLALVPVLTVMVAVASIF orf144a. pep PVFDRWSDSFVSFVNQTIVPQGADMVFDYINAFREQANRLTAIGSVMLWTSXMLIRTID I I II I I I I II II I I I II I II I II I I II I I I I II I I I I I I I I I I I I I I II I II I I II I II orf144-1 PVFDRWSDSFVSFVNQTIVPQGADMVFDYINAFREQANRLTAIGSVMLWTSLMLIRTID orf144a.pep NTFNRIWRVNSQRPWMMQFLVYWALLTFGPLSLGVGISFXVGSVQDAALASGAPQWSGAL I II I II I I I I II I I I II I II I II I I II I I I I I II I I I I I I I I II I II I I II II II II I I orf144-1 NTFNRIWRVNSQRPWMMQFLVYWALLTFGPLSLGVGISFMVGSVQDAALASGAPQWSGAL orf144a . pep RTAATLXFMTLLLWGLYRXVPNRFVPARXAFVGALATAFCLETARSLFTWYMGNFDGYRS I I I II I : I I I II I I I II I I I II I I I I I I I I II I I I I I II I I II I I I I II II II II II I orf144-1 RTAATLTFMTLLLWGLYRFVPNRFVPARQAFVGALATAFCLETARSLFTWYMGNFDGYRS orf144a. pep IYGAFAAVPFFLLWLNLLWTLVLGGAVLTSSLSYWQGEAFRRXFDSRGRFDDVLKILLLL II I I I II II II I I I I I I I I I I II I I I I I I I I I I II I I I I II I II II I I II I II II II II orf144-1 IYGAFAAVPFFLLWLNLLWTLVLGGAVLTSSLSYWQGEAFRRGFDSRGRFDDVLKILLLL orf144a. pep DAAQKEGXALPVQEFRRHINMGYDELGELLEKLARHGYIYSGRQGWVLKTGADSIELNEL I I II II I II I II I II II I I I I I I I II I I I I II I I I I I I II I I II I II I II II II II II I orf144-1 DAAQKEGKALPVQEFRRHINMGYDELGELLEKLARHGYIYSGRQGWVLKTGADSIELNEL orfl44a.pep FKLFVYRPLPVERDHVNQAVDAVMMPCLQTLNMTLAEFDAQAKKQQQS 408 I I I I I I I I II II I II I I I I I I I I I I I II I II II I I I I I I I II I: I orfl44-l FKLFVYRPLPVERDHVNQAVDAVMTPCLQTLNMTLAEFDAQAKKRQ 406 Homology with a predicted ORF from N.sonorrhoeae ORF144 shows 91.2% identity over a 136aa overlap with a predicted ORF (ORF144ng) from N.gonorrhoeae: orf144.pep MTFLQRLQGLADNKICAFAWFWRRFDEERVPQXAASMTFTTLLALVPVLTVMVAVASIF 60 II II I II II II I I II II I I : II I : I II I I I II I I I I I I II I II I I II II II I II I I orf144ng MTFLQCWQGSADNKICAFAWFVIRRFSEERVPQAAASMTFTTLLALVPVLTVMVAVASIF 60 orf144.pep PVFDRWSDSFVSFVNQTIVPXGADMVFDYINAFREQANRLTAIGSVMLWTSLMLIRTID 120 II II I II I II II II I I II II I I I II I I II: I II : I I I I I I II I I I I I I I I I I I I I I I I I orf144ng PVFDRWSDSFVSFVNQTIVPQGADMVFDYIDAFRDQANRLTAIGSVMLWTSLMLIRTID 120 orf144. pep NTFNRIWRVXXQRPWM 136 I : I I II I II MMM orfl44ng NAFNRIWRVNTQRPWMMQFLVYWALLTFGPLSLGVGISFMVGSVQDSVLSSGAQQWADAL 180 The complete length ORF144ng nucleotide sequence <SEQ ID 625> is predicted to encode a protein having amino acid sequence <SEQ ID 626>: 1 MTFLQCWQGS ADNKICAFAW FVIRRFSEER VPQAAASMTF TTLLALVPVL 51 TVMVAVASIF PVFDRWSDSF VSFVNQTIVP QGADMVFDYI DAFRDQANRL 101 TAIGSVMLW TSLMLIRTID NAFNRIWRVN TQRPWMMQFL VYWALLTFGP 151 LSLGVGISFM VGSVQDSVLS SGAQQWADAL KTAARLAFMT LLLWGLYRFV 201 PNRFVPARQA FVGALITAFC LETARFLFTW YMGNFDGYRS IYGAFAAVPF 251 FLLWLNLLWT LVLGGAVLTS SLSYWQGEAF RRGFDSRGRF DDVLKILLLL 301 DAAQKEGRTL SVQEFRRHIN MGYDELGELL EKLARYGYIY SGRQGWVLKT 351 GADSIELSEL FKLFVYRPLP VERDHVNQAV DAVMTPCLQT LNMTLAEFDA 401 QAKKQQQS* Further work revealed the following gonococcal DNA sequence <SEQ ED 627>: 1 ATGACCTTTT TACAACGTTG GCAAGGTTTG GCGGACAATA AAATCTGTGC 51 ATTTGCATGG TTCGTCATCC GCCGTTTCAG TGAAGAGCGC GTACCGCAGG 101 CAGCGGCGAG CATGACGTTT ACGACACTGC TGGCACTCGT CCCCGTACTG 151 ACCGTAATGG TCGCGGTCGC TTCGATTTTC CCCGTGTTCG ACCGCTGGTC 201 GGATTCGTTC GTCTCCTTCG TCAACCAAAC CATTGTGCCG CAGGGCGCGG 251 ATATGGTGTT CGACTATATC GACGCATTCC GCGATCAGGC AAACCGGCTG 301 ACCGCCATCG GCAGCGTGAT GCTGGTCGTA ACCTCGCTGA TGCTGATTCG 351 GACGATAGAC AATGCGTTCA ACCGCATCTG GCGGGTTAAC ACGCAACGCC 401 CCTGGATGAT GCAGTTCCTC GTTTATTGGG CGTTGCTGAC TTTCGGGCCT 451 TTGTCTTTGG GTGTGGGCAT TTCCTTTATG GTCGGGTCGG TTCAAGACTC 501 CGTACTCTCC TCCGGAGCGC AACAATGGGC GGACGCGTTG AAGACGGCGG 551 CAAGGCTGGC TTTCATGACG CTTTTGCTGT GGGGGCTGTA CCGCTTCGTG 601 CCCAACCGCT TCGTGCCCGC CCGGCAGGCG TTTGTCGGAG CTTTGATTAC 651 GGCATTCTGC CTGGAGACGG CACGTTTCCT GTTCACCTGG TATATGGGCA 701 ATTTCGACGG CTACCGCTCG ATTTACGGCG CATTTGCCGC CGTGCCGTTT 751 TTCCTGCTGT GGTTAAACCT GCTGTGGACG CTGGTCTTGG GCGGGGCGGT 801 GCTGACTTCG TCGCTGTCTT ATTGGCAGGG CGAGGCCTTC CGCAGGGGAT 851 TCGACTCGCG CGGACGGTTT GACGACGTGT TGAAAATCCT GCTGCTTCTG 901 GATGCGGCGC AAAAAGAAGG CCGAACCCTG TCCGTTCAGG AGTTCAGACG 951 GCATATCAAT ATGGGTTACG ATGAATTGGG CGAGCTTTTG GAAAAGCTGG 1001 CGCGGTACGG CTATATCTAT TCCGGCAGAC AGGGCTGGGT TTTGAAAACG 1051 GGGGCGGATT CGATTGAGTT GAGCGAACTC TTCAAGCTCT TCGTGTACCG 1101 CCCGTTGCct gtggaAAGGG ATCATGTGAA CCAAGCTGtc gaTGCGGTAA 1151 TGAcgccgtG TTTGCAGACT TTGAACATGA CGCTGGCGGA GTTTGACGCT 1201 CAGgcgAAAA AACAGCAGCA GTCTTGA This encodes a variant of ORF144ng, having the amino acid sequence <SEQ ID 628; ORF144ng-l>: 1 MTFLQRWQG ADNKICAFAW FVIRRFSEER VPQAAASMTF TTLLALVPVL 51 TVMVAVASIF PVFDRWSDSF VSFVNQTIVP QGADMVFDYI DAFRDQANRL 101 TAIGSVMLW TSLMLIRTID NAFNRIWRVN TQRPWMMQFL VYWALLTFGP 151 LSLGVGISFM VGSVQDSVLS SGAQQWADAL KTAARLAFMT LLLWGLYRFV 201 PNRFVPARQA FVGALITAFC LETARFLFTW YMGNFDGYRS IYGAFAAVPF 251 FLLWLNLLWT LVLGGAVLTS SLSYWQGEAF RRGFDSRGRF DDVLKILLLL 301 DAAQKEGRTL SVQEFRRHIN MGYDELGELL EKLARYGYIY SGRQGWVLKT 351 GADSIELSEL FKLFVYRPLP VERDHVNQAV DAVMTPCLQT LNMTLAEFDA 401 QAKKQQQS* ORF144ng-l and ORF144-1 show 94.1% identity in 406 aa overlap: orfl44ng-l . pep MTFLQRWQGLADNKICAFAWFVIRRFSEERVPQAAASMTFTTLLALVPVLTVMVAVASIF I I I I I I I I I I I I I I I I I I I I I : I I I : I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I orf 14 - 1 MTFLQRLQGLADNKICAFAWFWRRFDEERVPQAAASMTFTTLLALVPVLTVMVAVASIF orf l 44ng- l . pep PVFDRWSDSFVSFVNQTIVPQGADMVFDYIDAFRDQANRLTAIGSVMLWTSLMLIRTID I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I : I I I : I I I I I I I I I I I I I I I I I I I I I I I I I orf144-1 PVFDRWSDSFVSFVNQTIVPQGADMVFDYINAFREQANRLTAIGSVMLWTSLMLIRTID o fl44ng-l.pep NAFNRIWRVNTQRPWMMQFLVYWALLTFGPLSLGVGISFMVGSVQDSVLSSGAQQWADAL I : I I II I II I : I II II II I I I II II I I I I I II I I I I I II II I I I II :: I: I II I II orf144-1 NTFNRIWRVNSQRPWMMQFLVYWALLTFGPLSLGVGISFMVGSVQDAALASGAPQWSGAL orf144ng-l . pep KTAARLAFMTLLLWGLYRFVPNRFVPARQAFVGALITAFCLETARFLFTWYMGNFDGYRS M I : I I I I II I I I I I II II I II II I I I II I II I II I I I II I I I I I I II I I I I I I I orf144-1 RTAATLTFMTLLLWGLYRFVPNRFVPARQAFVGALATAFCLETARSLFTWYMGNFDGYRS orf144ng-l .pep IYGAFAAVPFFLLWLNLLWTLVLGGAVLTSSLSYWQGEAFRRGFDSRGRFDDVLKILLLL I I I I I II II I II I I I II II II I II I I I II II I I II II I I I II II II I I I I I II I I I I I II orf144-1 IYGAFAAVPFFLLWLNLLWTLVLGGAVLTSSLΞYWQGEAFRRGFDSRGRFDDVLKILLLL orfl44ng-l.pep DAAQKEGRTLSVQEFRRHINMGYDELGELLEKLARYGYIYSGRQGWVLKTGADSIELSEL II II II I :: I I I I I I I II I I I II I I I II II I I II : I I I I II II II I I I I I II I I I I : II orf144-1 DAAQKEGKALPVQEFRRHINMGYDELGELLEKLARHGYIYSGRQGWVLKTGADSIELNEL orf144ng-l . pep FKLFVYRPLPVERDHVNQAVDAVMTPCLQTLNMTLAEFDAQAKKQQQS I II I I I I II I I I I I I I I II I I I I I I I I II II I I I I I I I I I II II : I orfl44-l FKLFVYRPLPVERDHVNQAVDAVMTPCLQTLNMTLAEFDAQAKKRQ On this basis of this analysis, including the identification of several putative transmembrane domains in the gonococcal protein, it is predicted that the proteins from N. meningitidis and N.gonorrhoeae, and their epitopes, could be useful antigens for vaccines or diagnostics, or for raising antibodies. Example 75 The following partial DNA sequence was identified in N meningitidis <SEQ ID 629>: 1 ..AGACACGCCC GCCGCATCCG CATCGACACC GCCATCAACC CCGAACTGGA 51 AGCCCTCGCC GAACACCTCC ACTACCAATG GCAGGGCTTC CTCTGGCTCA 101 GCACCGATAT GCGTCAGGAA ATTTCCGCCC TCGTCATCCT GCTGCAACGC 151 ACCCGCCGCA AATGGCTGGA TGCCCACGAA CGCCAACACC TGCGCCAAAG 201 CCTGCTTGAA ACACGGGAAC ACGGCTGA This coπesponds to the amino acid sequence <SEQ ID 630; ORF146>: 1 ..RHARRIRIDT AIΝPELEALA EHLHYQWQGF LWLSTDMRQE ISALVILLQR 51 TRRKWLDAHE RQHLRQSLLE TREHG* Further work revealed the complete nucleotide sequence <SEQ ID 631>: 1 ATGAACACCT CGCAACGCAA CCGCCTCGTC AGCCGCTGGC TCAACTCCTA 51 CGAACGCTAC CGCTACCGCC GCCTCATCCA CGCCGTCCGG CTCGGCGGGG 101 CCGTCCTGTT CGCCACCGCC TCCGCCCGGC TGCTCCACCT CCAACACGGC 151 GAGTGGATAG GGATGACCGT CTTCGTCGTC CTCGGCATGC TCCAGTTTCA 201 AGGGGCGATT TACTCCAAGG CGGTGGAACG TATGCTCGGC ACGGTCATCG 251 GGCTGGGCGC GGGTTTGGGC GTTTTATGGC TGAACCAGCA TTATTTCCAC 301 GGCAACCTCC TCTTCTACCT CACCGTCGGC ACGGCAAGCG CACTGGCCGG 351 CTGGGCGGCG GTCGGCAAAA ACGGCTACGT CCCTATGCTG GCAGGGCTGA 401 CGATGTGTAT GCTCATCGGC GACAACGGCA GCGAATGGCT CGACAGCGGA 451 CTCATGCGCG CCATGAACGT CCTCATCGGC GCGGCCATCG CCATCGCCGC 501 CGCCAAACTG CTGCCGCTGA AATCCACACT GATGTGGCGT TTCATGCTTG 551 CCGACAACCT GGCCGACTGC AGCAAAATGA TTGCCGAAAT CAGCAACGGC 601 AGGCGCATGA CCCGCGAACG CCTCGAGGAG AACATGGCGA AAATGCGCCA 651 AATCAACGCA CGCATGGTCA AAAGCCGCAG CCATCTCGCC GCCACATCGG 701 GCGAAAGCCG CATCAGCCCC GCCATGATGG AAGCCATGCA GCACGCCCAC 751 CGTAAAATCG TCAACACCAC CGAGCTGCTC CTGACCACCG CCGCCAAGCT 801 GCAATCTCCC AAACTCAACG GCAGCGAAAT CCGGCTGCTT GACCGCCACT 851 TCACACTGCT CCAAACCGAC CTGCAACAAA CCGTCGCCCT TATCAACGGC 901 AGACACGCCC GCCGCATCCG CATCGACACC GCCATCAACC CCGAACTGGA 995511 AGCCCTCGCC GAACACCTCC ACTACCAATG GCAGGGCTTC CTCTGGCTCA 1001 GCACCAATAT GCGTCAGGAA ATTTCCGCCC TCGTCATCCT GCTGCAACGC 1051 ACCCGCCGCA AATGGCTGGA TGCCCACGAA CGCCAACACC TGCGCCAAAG 1101 CCTGCTTGAA ACACGGGAAC ACGGCTGA This coπesponds to the amino acid sequence <SEQ ID 632; ORF146-l>: 1 MNTSQRNRLV SRWLNSYERY RYRRLIHAVR LGGAVLFATA SARLLHLQHG 51 EWIGMTVFW LGMLQFQGAI YSKAVERMLG TVIGLGAGLG VLWLNQHYFH 101 GNLLFYLTVG TASALAGWAA VGKNGYVPML AGLTMCMLIG DNGSEWLDSG 151 LMRAMNVLIG AAIAIAAAKL LPLKSTLMWR FMLADNLADC SKMIAEISNG 201 RRMTRERLEE NMAKMRQINA RMVKSRSHLA ATSGESRISP AMMEAMQHAH 251 RKIVNTTELL LTTAAKLQSP KLNGSEIRLL DRHFTLLQTD LQQTVALING 301 RHARRIRIOΥ AINPELEALA EHLHYQWQGF LWLSTNMRQE ISALVILLQR 351 TRRKWLDAHE RQHLRQSLLE TREHG* Computer analysis of this amino acid sequence gave the following results: Homology with a predicted ORF from Nmeningitidis (strain A) ORF146 shows 98.6% identity over a 74aa overlap with an ORF (ORF146a) from sfrain A of N meningitidis: 10 20 30 orf146. pep RHARRIRIDTAIΝPELEALAEHLHYQWQGF I II I II I I I II II I I I I I I I I I I I II II II orf146a KLΝGSEIRLLDRHFTLLQTDLQQTVALIΝGRHARRIRIDTAIΝPELEALAEHLHYQWQGF 280 290 300 310 320 330 40 50 60 70 orf146. pep LWLSTDMRQEISALVILLQRTRRKWLDAHERQHLRQSLLETREHGX II I II : II II II II I II II I I I II I II I I I II I II I I I II I I II : orf146a LWLSTΝMRQEISALVILLQRTRRKWLDAHERQHLRQSLLETREHSX 340 350 360 370 The complete length ORF 146a nucleotide sequence <SEQ ID 633> is: 1 ATGAACACCT CGCAACGCAA CCGCCTCGTC AGCCGCTGGC TCAACTCCTA 51 CGAACGCTAC CGCTACCGCC GCCTCATCCA CGCCGTCCGG CTCGGCGGGG 101 CCGTCCTGTT CGCCACCGCC TCCGCCCGGC TGCTCCACCT CCAACACGGC 151 GAGTGGATAG GGATGACCGT CTTCGTCGTC CTCGGCATGC TCCAGTTTCA 201 AGGGGCGATT TACTCCAAGG CGGTGGAACG TATGCTCGGC ACGGTCATCG 251 GGCTGGGCGC GGGTTTGGGC GTTTTATGGC TGAACCAGCA TTATTTCCAC 301 GGCAACCTCC TCTTCTACCT CACCGTCGGC ACGGCAAGCG CACTGGCCGG 351 CTGGGCGGCG GTCGGCAAAA ACGGCTACGT CCCTATGCTG GCGGGGCTGA 401 CGATGTGCAT GCTCATCGGC GACAACGGCA GCGAATGGTT CGACAGCGGC 451 CTGATGCGCG CGATGAACGT CCTCATCGGC GCGGCCATCG CCATCGCCGC 501 CGCCAAACTG CTGCCGCTGA AATCCACACT GATGTGGCGT TTCATGCTTG 551 CCGACAACCT GACCGACTGC AGCAAAATGA TTGCCGAAAT CAGCAACGGC 601 AGGCGCATGA CCCGCGAACG CCTCGAAGAG AACATGGCGA AAATGCGCCA 651 AATCAACGCA CGCATGGTCA AAAGCCGCAG CCACCTCGCC GCCACATCGG 701 GCGAAAGCCG CATCAGCCCC GCCATGATGG AAGCCATGCA GCACGCCCAC 751 CGTAAAATTG TCAACACCAC CGAGCTGCTC CTGACCACCG CCGCCAAGCT 801 GCAATCTCCC AAACTCAACG GCAGCGAAAT CCGGCTGCTT GACCGCCACT 851 TCACACTGCT CCAAACCGAC CTGCAACAAA CCGTCGCCCT TATCAACGGC 901 AGACACGCCC GCCGCATCCG CATCGACACC GCCATCAACC CCGAACTGGA 951 AGCCCTCGCC GAACACCTCC ACTACCAATG GCAGGGCTTC CTCTGGCTCA 1001 GCACCAATAT GCGTCAGGAA ATTTCCGCCC TCGTCATCCT GCTGCAACGC 1051 ACCCGCCGCA AATGGCTGGA TGCCCACGAA CGCCAACACC TGCGCCAAAG 1101 CCTGCTTGAA ACACGGGAAC ACAGTTGA This encodes a protein having amino acid sequence <SEQ ID 634>: 1 MNTSQRNRLV SRWLNSYERY RYRRLIHAVR LGGAVLFATA SARLLHLQHG 51 EWIGMTVF LGMLQFQGAI YSKAVERMLG TVIGLGAGLG VLWLNQHYFH 101 GNLLFYLTVG TASALAGWAA VGKNGYVPML AGLTMCMLIG DNGSEWFDSG 151 LMRAMNVLIG AAIAIAAAKL LPLKSTLMWR FMLADNLTDC SKMIAEISNG 201 RRMTRERLEE NMAKMRQINA RMVKSRSHLA ATSGESRISP AMMEAMQHAH 251 RKIVNTTELL LTTAAKLQSP KLNGSEIRLL DRHFTLLQTD LQQTVALING 301 RHARRIRIDT AINPELEALA EHLHYQWQGF LWLSTNMRQE ISALVILLQR 351 TRRKWLDAHE RQHLRQSLLE TREHS* ORF146a and ORF146-1 show 99.5% identity in 374 aa overlap: orf146a . pep MNTSQRNRLVSRWLNSYERYRYRRLIHAVRLGGAVLFATASARLLHLQHGEWIGMTVFW I I I I I II I I II II I I I I II I I II I II II II I II I I I I I I I I I I II I I I II II I I I II I I I orf146-1 MNTSQRNRLVSRWLNSYERYRYRRLIHAVRLGGAVLFATASARLLHLQHGEWIGMTVFW orf146a . pep LGMLQFQGAIYSKAVERMLGTVIGLGAGLGVLWLNQHYFHGNLLFYLTVGTASALAGWAA I I I II I I II I II I I I I II I I II I II I I I I I II I II I I I II I I I I II I I II I I I II II I I I orf146-1 LGMLQFQGAIYSKAVERMLGTVIGLGAGLGVLWLNQHYFHGNLLFYLTVGTASALAGWAA orf146a. pep VGKNGYVPMLAGLTMCMLIGDNGSEWFDSGLMRAMNVLIGAAIAIAAAKLLPLKSTLMWR I II I II I I II II I I I I I II I I II I II : II II II I II I II II I II I II II I I II II I I II I orf146-1 VGKNGYVPMLAGLTMCMLIGDNGSEWLDSGLMRAMNVLIGAAIAIAAAKLLPLKSTLMWR orf146a. pep FMLADNLTDCSKMIAEISNGRRMTRERLEENMAKMRQINARMVKSRSHLAATSGESRISP I I I I I I I II I I II II I II I I II I I I I I I II I II I I I I I II I II I I I II II I I I II I I I orf146-1 FMLADNLADCSKMIAEISNGRRMTRERLEENMAKMRQINARMVKSRSHLAATΞGESRISP orf146a. pep AMMEAMQHAHRKIVNTTELLLTTAAKLQSPKLNGSEIRLLDRHFTLLQTDLQQTVALING I II I II I I I I I II I I I I I II I I II I I I I I I II I II I I I II II I II I I I II II I I I II I I I orfl46-l AMMEAMQHAHRKIVNTTELLLTTAAKLQSPKLNGSEIRLLDRHFTLLQTDLQQTVALING orf146a. pep RHARRIRIDTAINPELEALAEHLHYQWQGFLWLSTNMRQEISALVILLQRTRRKWLDAHE II I I I I II I I I I I II I II I I I II I II II II I II I II I II I I I II I I I I I II I II II I I II orfl46-l RHARRIRIDTAINPELEALAEHLHYQWQGFLWLSTNMRQEISALVILLQRTRRKWLDAHE orf146a. pep RQHLRQSLLETREHSX I I I II I I II I II I I: orf146-1 RQHLRQSLLETREHGX Homology with a predicted ORF from N.sonorrhoeae ORF146 shows 97.3% identity over a 75aa overlap with a predicted ORF (ORF146ng) from N.gonorrhoeae: orf146. pep RHARRIRIDTAINPELEALAEHLHYQWQGF 30 II I II I I I I II II I I I I I I I I I I I I II II I orfl46ng KLNGSEIRLLDRHFTLLQTDLQQTAALINGRHARRIRIDTAINPELEALAEHLHYQWQGF 364 orf146. pep LWLSTDMRQEISALVILLQRTRRKWLDAHERQHLRQSLLETREHG 75 I I I I IM I II I I II I I II I II I I II I I I I II I II I II II I II I I orfl46ng LWLSTNMRQEISALVIPLQRTRRKWLDAHERQHLRQSLLETREHG 409 An ORF146ng nucleotide sequence <SEQ LD 635>was predicted to encode aprotein having amino acid sequence <SEQ ID 636>: 1 MSGVRFPSPA PIPSTDPPSG SLCFFTFPLQ TASDMWSSQR KRLSGRWLNS 51 YERYRHRRLI HAVRLGGTVL FATALARLLH LQHGEWIGMT VFWLGMLQF 101 QGAIYSNAVE RMLGTVIGLG AGLGVLWLNQ HYFHGNLLFY LTIGTASALA 151 GWAAVGKNGY VPMLAGLTMC MLIGDNGSEW LDSGLMRAMN VLIGAAIAIA 201 AAKLLPLKST LMWRFMLADN LADCSKMIAE ISNGRRMTRE RLEQNMVKMR 251 QINARMVKSR SHLAATSGES RISPSMMEAM QHAHRKIVNT TELLLTTAAK 301 LQSPKLNGSE IRLLDRHFTL LQTDLQQTAA LINGRHARRI RIDTAINPEL 351 EALAEHLHYQ WQGFLWLSTN MRQEISALVI PLQRTRRKWL DAHERQHLRQ 401 SLLETREHG* Further work revealed the following gonococcal DNA sequence <SEQ LD 637>: 1 ATGAACTCCT CGCAACGCAA ACGCCTTTCC GgccGCTGGC TCAACTCCTA 51 CGAACGCTac cGCCaccGCC GCCTCATACA TGCCGTGCGG CTCGGCggaa 101 ccgtCCTGTT CGCCACCGCA CTCGCCCGgc tACTCCACCT CCAacacggc 151 gAATGGATAG GGAtgaCCGT CTTCGTCGTC CTCGGCATGC TCCAGTTCCA 201 AGGCgcgatt tActccaacg cggtgGAacg taTGctcggt acggtcatcg 251 ggctgGGCGC GGGTTTGGgc gTTTTATGGC TGAACCAGCA TTAtttccac 301 ggcaacCTcc tcttctacct gaccatcggc acggcaagcg cactggccgg 351 ctGGGCGGCG GTCGGCAAAA acggctacgt ccctatgctg GCGGGGctgA 401 CGATGTGCAT gctcatcggc gACAACGGCA GCGAATGGCT CGACAGCGGC 451 CTGATGCGCG CGATGAACGT CCTCATCGGC GCCGCCATCG CCATTGCCGC 501 CGCCAAACTG CTGCCGCTGA AATCCACACT GATGTGGCGT TTCATGCTTG 551 CCGACAACCT GGCCGACTGC AGCAAAATGA TTGCCGAAAT CAGCAACGGC 601 AGGCGTATGA CGCGCGAACG TTTGGAGCAG AATATGGTCA AAATGCGCCA 651 AATCAACGCA CGCATGGTCA AAAGCCGCAG CCACCTCGCC GCCACATCGG 701 GCGAAAGCCG CATCAGCCCC TCCATGATGG AAGCCATGCA GCACGCCCAC 751 CGCAAAATCG TCAACACCAC CGAGCTGCTC CTGACCACCG CCGCCAAGCT 801 GCAATCTCCC AAACTCAACG GCAGCGAAAT CCGGCTGCTC GACCGCCACT 851 TCACACTGCT CCAAACCGAC CTGCAACAAA CCGCCGCCCT CATCAACGGC 901 AGACACGCCC GCCGCATCCG CATCGACACC GCCATCAACC CCGAACTGGA 951 AGCCCTCGCC GAACACCTCC ACTACCAATG GCAGGGCTTC CTCTGGCTCA 1001 GCACCAATAT GCGTCAGGAA ATTTCCGCCC TCGTCATCCT GCTGCAACGC 1051 ACCCGCCGCA AATGGCTGGA TGCCCACGAA CGCCAACACC TGCGCCAAAG 1101 CCTGCTTGAA ACACGGGAAC ACGGCTGA This coπesponds to the amino acid sequence <SEQ ID 638; ORF146ng-l>: 1 MNSSQRKRLS GRWLNSYERY RHRRLIHAVR LGGTVLFATA LARLLHLQHG 51 EWIGMTVFW LGMLQFQGAI YSNAVERMLG TVIGLGAGLG VLWLNQHYFH 101 GNLLFYLTIG TASALAGWAA VGKNGYVPML AGLTMCMLIG DNGSEWLDSG 151 LMRAMNVLIG AAIAIAAAKL LPLKSTLMWR FMLADNLADC SKMIAEISNG 201 RRMTRERLEQ NMVKMRQINA RMVKSRSHLA ATSGESRISP SMMEAMQHAH 251 RKIVNTTELL LTTAAKLQSP KLNGSEIRLL DRHFTLLQTD LQQTAALING 301 RHARRIRIDT AINPELEALA EHLHYQWQGF LWLSTNMRQE ISALVILLQR 351 TRRKWLDAHE RQHLRQSLLE TREHG* ORF146ng-l and ORF146-1 show 96.5% identity in 375 aa overlap orf146-1. pep MNTSQRNRLVSRWLNSYERYRYRRLIHAVRLGGAVLFATASARLLHLQHGEWIGMTVFW II: I II : I I : II II I II II I : II I II I II I I I : I II I I I I II I I I I I II I I I I I I II I orfl46ng-l MNSSQRKRLSGRWLNSYERYRHRRLIHAVRLGGTVLFATALARLLHLQHGEWIGMTVFW orfl46-l.pep LGMLQFQGAIYSKAVERMLGTVIGLGAGLGVLWLNQHYFHGNLLFYLTVGTASALAGWAA I I I I II I II II I : II I II I I II II I I II I II I I I II I II I I I I I I I I I : I II II II I I I I orf146ng-l LGMLQFQGAIYSNAVERMLGTVIGLGAGLGVLWLNQHYFHGNLLFYLTIGTASALAGWAA orf146-1.pep VGKNGYVPMLAGLTMCMLIGDNGSEWLDSGLMRAMNVLIGAAIAIAAAKLLPLKSTLMWR I I I I II II II II I II I I I I II I II I II I II I I I II II I I I II II I I I I II II I I II I I II orfl46ng-l VGKNGYVPMLAGLTMCMLIGDNGSEWLDSGLMRAMNVLIGAAIAIAAAKLLPLKSTLMWR orfl46-l .pep FMLADNLADCSKMIAEISNGRRMTRERLEENMAKMRQINARMVKSRSHLAATSGESRISP I I I II I I I I I I I I I I II I I I I II I I I I I I : I I: I I I I I II II II II I I I II I I I I I I II I orf146ng-l FMLADNLADCSKMIAEISNGRRMTRERLEQNMVKMRQINARMVKSRSHLAATSGESRISP orfl46-l.pep AMMEAMQHAHRKIVNTTELLLTTAAKLQSPKLNGSEIRLLDRHFTLLQTDLQQTVALING : I I I II I II I I I I I I II I I I I II I I I I I I I I I II I I I I I I II II II I I II I I I I: I I I II orfl46ng-l SMMEAMQHAHRKIVNTTELLLTTAAKLQSPKLNGSEIRLLDRHFTLLQTDLQQTAALING orf146-1.pep RHARRIRIDTAINPELEALAEHLHYQWQGFLWLSTNMRQEISALVILLQRTRRKWLDAHE I I II I I I I II II I II I II I I II I I II I II I II II I I I I I I I I I I I I I I I II I I I I I II I I orfl46ng-l RHARRIRIDTAINPELEALAEHLHYQWQGFLWLSTNMRQEISALVILLQRTRRKWLDAHE orf146-1.pep RQHLRQSLLETREHGX I I I I II I II I I I I I I I orf146ng-l RQHLRQSLLETREHGX Furthermore, ORF146ng-l shows homology with a hypothetical E.coli protein: sp|P33011|YEEA_ECOLI HYPOTHETICAL 40.0 KD PROTEIN IN COBU-SBMC INTERGENIC REGION >gi|173667 | gnl | PID I dl016553 (D90838) ORF_ID:o348#20; similar to [SwissProt Accession Number P33011] [Escherichia coli] >gi 11736682 | gnl | PID| dl016560 (D90839) ORF ID:o348#20; similar to [SwissProt Accession Number P33011] [Escherichia coli] >gi 11788318 (AE000292) f352; 100% identical to fragment YEEA_ECOLI SW: P33011 but has 203 additional C-terminal residues [Escherichia coli] Length = 352 Score = 109 bits (271), Expect = 2e-23 Identities = 89/347 (25%), Positives = 150/347 (42%), Gaps = 21/347 (6%) Query: 20 YRHRRLIHAVRLGGTVLFATALARLLHLQHGEWIGMTVFWLGMLQFQGAIYSNAVERML 79 YRH R++H R+ L + RL + W +T+ V++G + F G + A ER+ Sbjct: 15 YRHYRIVHGTRVALAFLLTFLIIRLFTIPESTWPLVTMWIMGPISFWGNWPRAFERIG 74 Query: 80 GTVIGLGAGLGVLWLNQHYFHGNLLFYLTIGTASALAGWAAVGKNGYVPMLAGLTMCMLI 139 GTV+G GL L L L + A L GW A+GK Y +L G+T+ +++ Sbjct: 75 GTVLGSILGLIALQLE LISLPLMLVWCAAAMFLCGWLALGKKPYQGLLIGVTLAIW 131 Query: 140 GDNGSEWLDSGLMRAMNVLIGXXXXXXXXKLLPLKSTLMWRFMLADNLADCSKMIAEISN 199 G E +D+ L R+ +V++G + P ++ + WR LA +L + +++ + Sbjct: 132 GSPTGE-IDTALWRSGDVILGSLLAMLFTGIWPQRAFIHWRIQLAKSLTEYNRVYQSAFS 190 Query: 200 GRRMTRERLEQNMVKMRQINARMVKSRSHLAATSGESRISPSMMEAMQHAHRKIVNXXXX 259 + R RLE ++ K+ VK R +A S E+RI S+ E +Q +R +V Sbjct: 191 PNLLERPRLESHLQKLL TDAVKMRGLIAPASKETRIPKSIYEGIQTINRNLVCMLEL 247 Query: 260 XXXXXXXXQSPK LNGSEIRLLDRHFXXXXXXXXXXAALINGRHARRIRIDTAINPEL 316 + LN ++R D AL G +N + Sbjct: 248 QINAYWATRPSHFVLLNAQKLR—DTQHMMQQILLSLVHALYEGNPQPVFANTEKLNDAV 305 Query: 317 EALAEHL—HYQWQ GFLWLSTNMRQEISALVILLQRTRRK 354 E L + L H+ + G++WL+ ++ L L+ R RK Sbjct: 306 EELRQLLNNHHDLKWETPIYGYVWLNMETAHQLELLSNLICRALRK 352 On the basis of this analysis, including the identification of several fransmembrane domains in the gonococcal protein, it is predicted that the proteins from N. meningitidis and N.gonorrhoeae, and their epitopes, could be useful antigens for vaccines or diagnostics, or for raising antibodies. Example 76 The following partial DNA sequence was identified in N. meningitidis <SEQ ID 639> 1 .. GCCGAAGACA CGCGCGTTAC CGCACAGCTT TTGAGCGCGT ACGGCATTCA 51 GGGCAAACTC GTCAGTGTGC GCGAACACAA CGAACGGCAG ATGGCGGACA 101 AGATTGTCGG CTATCTTTCA GACGGCATGG TTGTGGCACA GGTTTCCGAT 151 GCGGGTACGC CGGCCGTGTG CGACCCGGGC GCGAAACTCG CCCGCCGCGT 201 GCGTGAGGCC GGGTTTAAAG TCGTTCCCGT CGTGGGCGCA AC . GCGGTGA 251 TGGCGGCTTT GAGCGTGGCC GGTGTGGAAG GATCCGATTT TTATTTCAAC 301 GGTTTTGTAC CGCCGAAATC GGGAGAACGC AGGAAACTGT TTGCCAAATG 351 GGTGCGGGCG GCGTTTCCTA TCGTCATGTT TGAAACGCCG CACCGCATCG 401 GTGCAGCGCT TGCCGATATG GCGGAACTGT TCCCCGAACG CCGATTAATG 451 CTGGCGCGCG AAATTACGAA AACGTTTGAA ACGTTCTTAA GCGGCACGGT 501 TGGGGAAATT CAGACGGCAT TGTCTGCCGA CGGCGACCAA TCGCGCGGCG 551 AGATGGTGTT GGTGCTTTAT CCGGCGCAGG ATGAAAAACA CGAAGGCTTG 601 TCCGAGTCCG CGCAAAACAT CATGAAAATC CTCACAGCCG AGCTGCCGAC 651 CAAACAGGCG GCGGAGCTTG CTGCCAAAAT CACGGGCGAG GGAAAGAAAG 701 CTTTGTACGA T.. This coπesponds to the .amino acid sequence <SEQ ID 640; ORF147>: 1 ..AEDTRVTAQL LSAYGIQGKL VSVREHNERQ MADKIVGYLS DGMWAQVSD 51 AGTPAVCDPG AKLARRVREA GFKWPWGA XAVMAALSVA GVEGSDFYFN 101 GFVPPKSGER RKLFAKWVRA AFPIVMFETP HRIGAALADM AELFPERRLM 151 LAREITKTFE TFLSGTVGEI QTALSADGDQ SRGEMVLVLY PAQDEKHEGL 201 SESAQNIMKI LTAELPTKQA AELAAKITGE GKKALYD .. Further work revealed the complete nucleotide sequence <SEQ ID 641>: 1 ATGTTTCAGA AACATTTGCA GAAAGCCTCC GACAGCGTCG TCGGAGGGAC 51 ATTATACGTG GTTGCCACGC CCATCGGCAA TTTGGCGGAC ATTACCCTGC 101 GCGCTTTGGC GGTATTGCAA AAGGCGGACA TCATCTGTGC CGAAGACACG 151 CGCGTTACCG CACAGCTTTT GAGCGCGTAC GGCATTCAGG GCAAACTCGT 201 CAGTGTGCGC GAACACAACG AACGGCAGAT GGCGGACAAG ATTGTCGGCT 251 ATCTTTCAGA CGGCATGGTT GTGGCACAGG TTTCCGATGC GGGTACGCCG 301 GCCGTGTGCG ACCCGGGCGC GAAACTCGCC CGCCGCGTGC GTGAGGCCGG 351 GTTTAAAGTC GTTCCCGTCG TGGGCGCAAG CGCGGTGATG GCGGCTTTGA 401 GCGTGGCCGG TGTGGAAGGA TCCGATTTTT ATTTCAACGG TTTTGTACCG 451 CCGAAATCGG GAGAACGCAG GAAACTGTTT GCCAAATGGG TGCGGGCGGC 501 GTTTCCTATC GTCATGTTTG AAACGCCGCA CCGCATCGGT GCGACGCTTG 551 CCGATATGGC GGAACTGTTC CCCGAACGCC GATTAATGCT GGCGCGCGAA 601 ATTACGAAAA CGTTTGAAAC GTTCTTAAGC GGCACGGTTG GGGAAATTCA 651 GACGGCATTG TCTGCCGACG GCAACCAATC GCGCGGCGAG ATGGTGTTGG 701 TGCTTTATCC GGCGCAGGAT GAAAAACACG AAGGCTTGTC CGAGTCCGCG 751 CAAAACATCA TGAAAATCCT CACAGCCGAG CTGCCGACCA AACAGGCGGC 801 GGAGCTTGCT GCCAAAATCA CGGGCGAGGG AAAGAAAGCT TTGTACGATC 851 TGGCTCTGTC TTGGAAAAAC AAATAG This coπesponds to the amino acid sequence <SEQ ID 642; ORF147-l>: 1 MFQKHLQKAS DSWGGTLYV VATPIGNLAD ITLRALAVLQ KAΌIICAEDT 51 RVTAQLLSAY GIQGKLVSVR EHNERQMADK IVGYLSDGMV VAQVSDAGTP 101 AVCDPGAKLA RRVREAGFKV VPWGASAVM AALSVAGVEG SDFYFNGFVP 151 PKSGERRKLF AKWVRAAFPI VMFETPHRIG ATLADMAELF PERRLMLARE 201 ITKTFETFLS GTVGEIQTAL SADGNQSRGE MVLVLYPAQD EKHEGLSESA 251 QNIMKILTAE LPTKQAAELA AKITGEGKKA LYDLALSWKN K* Computer analysis of this amino acid sequence gave the following results: Homology with hypothetical protein ORF286 of E.coli (accession number UI 89971 ORF147 and E.coli ORF286 protein show 36% aa identity in 237aa overlap: Orfl47 1 AEDTRVTAQLLSAYGIQGKLVSVREHNERQMADKIVGYLSDGMWAQVSDAGTPAVCDPG 60 AEDTR T LL +GI +L ++ +HNE+Q A+ ++ L +G +A VSDAGTP + DPG Orf286 43 AEDTRHTGLLLQHFGINARLFALHDHNEQQKAETLLAKLQEGQNIALVSDAGTPLINDPG 102 Orfl47 61 AKLARRVREXXXXXXXXXXXXXXXXXXXXXXXEGSDFYFNGFVPPKSGERRKLFAKWVRA 120 L R RE F + GF+P KS RR Orf286 103 YHLVRTCREAGIRWPLPGPCAAITALSAAGLPSDRFCYEGFLPAKΞKGRRDALKAIEAE 162 Orfl47 121 AFPIVMFETPHRIGAALADMAELFPERR-LMLAREITKTFETFLSGTVGEIQTALSADGD 179 ++ +E+ HR+ +L D+ + E R ++LARE+TKT+ET VGE+ + D + Orf286 163 PRTLIFYESTHRLLDSLEDIVAVLGESRYWLARELTKTWETIHGAPVGELLAWVKEDEN 222 Orfl47 180 QSRGEMVLVLYPAQDEKHEGLSESAQNIMKILTAELPTKQAAELAAKITGEGKKALY 236 + +GEMVL++ + E L A + +L AELP K+AA LAA+I G K ALY Orf286 223 RRKGEMVLIV-EGHKAQEEDLPADALRTLALLQAELPLKKAAALAAEIHGVKKNALY 278 Homology with a predicted ORF from N.meninsitidis (strain Al ORF147 shows 96.6% identity over a 237aa overlap with ORF75a from strain A of N meningitidis: 10 20 30 orf147.pep AEDTRVTAQLLSAYGIQGKLVSVREHΝERQ I I II II II I I I I I I I I I II I I I II I II I I I orf75a TLY ATPIGΝLADITLRALAVLQKADIICAEDTRVTAQLLSAYGIQGKLVSVREHΝERQ 20 30 40 50 60 70 40 50 60 70 80 90 orf147.pep MADKIVGYLSDGMWAQVSDAGTPAVCDPGAKLARRVREAGFKWPWGAXAVMAALSVA I I I I I I I I II I I II I II I II I I I II I II II I I I I I II II : I II I II II I I II I I I I II I orf75a MADKIVGYLSDGMWAQVSDAGTPAVCDPGAKLARRVREVGFKWPWGASAVMAALSVA 80 90 100 110 120 130 100 110 120 130 140 150 orf147.pep GVEGSDFYFΝGFVPPKSGERRKLFAKWVRAAFPIVMFETPHRIGAALADMAELFPERRLM II I I I I I II I I II I II I II I I I I I I I I I : I IM I I I I I I I I I IM I I I I II I I I I I I II orf75a GVAGSDFYFΝGFVPPKSGERRKLFAKWVRVAFPWMFETPHRIGATLADMAELFPERRLM 140 150 160 170 180 190 160 170 180 190 200 210 orf147.pep LAREITKTFETFLSGTVGEIQTALSADGDQSRGEMVLVLYPAQDEKHEGLSESAQΝIMKI I I I II I I I II II II I II II II I I I : I I I : II II I II I I I I I I II II I I I I I I I II II II I orf75a LAREITKTFETFLSGTVGEIQTALAADGNQSRGEMVLVLYPAQDEKHEGLSESAQNIMKI 200 210 220 230 240 250 220 230 orfl47.pep LTAELPTKQAAELAAKITGEGKKALYD I I I I I I I I I I I I II II I I II II I I I I I orf75a LTAELPTKQAAELAAKITGEGKKALYDLALSWKNKX 260 270 280 290 ORF147a is identical to ORF75a, which includes aa 56-292 of ORF75. Homology with a predicted ORF from N.sonorrhoeae ORF 147 shows 94.1% identity over a 237aa overlap with a predicted ORF (ORF147ng) from N gonorrhoeae: orf 147 . pep AEDTRVTAQLLSAYGIQGKLVSVREHNERQ 30 I I I I I I I I I I I I I I I I I I : I I I I I I I I I I I orfl47ng TLYWATPIGNLADITLRALAVLQKADIICAEDTRVTAQLLSAYGIQGRLVSVREHNERQ 85 orf 147 . pep MADKIVGYLSDGMWAQVSDAGTPAVCDPGAKLARRVREAGFKWPWGAXAVMAALSVA 90 I I I M : M I I I I : I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I orfl 47ng MADKVIGFLSDGLWAQVSDAGTPAVCDPGAKLARRVREAGFKWPWGASAVMAALSVA 145 orf 147 . pep GVEGSDFYFNGFVPPKSGERRKLFAKWVRAAFPIVMFETPHRIGAALADMAELFPERRLM 150 I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I : I I I I I I I I I I I I I I orfl 47ng GVAESDFYFNGFVPPKSGERRKLFAKWVRAAFPWMFETPHRIGATLADMAELFPERRLM 205 orf147.pep LAREITKTFETFLSGTVGEIQTALSADGDQSRGEMVLVLYPAQDEKHEGLSESAQNIMKI 210 II I I II II II II II I I I I I I I I I I : I I I : I II II I I I I II II I II II I I I II I I II III orfl47ng LAREITKTFETFLSGTVGEIQTALAADGNQSRGEMVLVLYPAQDEKHEGLSESAQNAMKI 265 orf147. pep LTAELPTKQAAELAAKITGEGKKALYD 237 I : I I I I I I I I II II I I I I I I I I I I II I orfl47ng LAAELPTKQAAELAAKITGEGKKALYDLALSWKNK 300 An ORF147ng nucleotide sequence <SEQ D 643> was predicted to encode a protein having amino acid sequence <SEQ ID 644>: 1 MSVFQTAFFM FQKHLQKASD SWGGTLYW ATPIGNLADI TLRALAVLQK 51 ADIICAEDTR VTAQLLSAYG IQGRLVSVRE HNERQMADKV IGFLSDGLW 101 AQVSDAGTPA VCDPGAKLAR RVREAGFKW PWGASAVMA ALSVAGVAES 151 DFYFNGFVPP KSGERRKLFA KWVRAAFPW MFETPHRIGA TLADMAELFP 201 ERRLMLAREI TKTFETFLSG TVGEIQTALA ADGNQSRGEM VLVLYPAQDE 251 KHEGLSESAQ NAMKILAAEL PTKQAAELAA KITGEGKKAL YDLALSWKNK 301 * Further work revealed the following gonococcal DNA sequence <SEQ ID 645>: 1 ATGTTTCAGA AACACTTGCA GAAAGCCTCC GACAGCGTCG TCGGAGGGAC 51 ATTATACGTG GTTGCCACGC CCATCGGCAA TTTGGCAGAC ATTACCCTGC 101 GCGCTTTGGC GGTATTGCAA AAGGCGGACA TCATTTGTGC CGAAGACACG 151 CGCGTTACTG CGCAGCTTTT GAGCGCGTAC GGCATTCAGG GCAGGTTGGT 201 CAGTGTGCGC GAACACAACG AGCGGCAGAT GGCGGACAAG GTAATCGGTT 251 TCCTTTCAGA CGGCCTGGTT GTGGCGCAGG TTTCCGATGC GGGTACGCCG 301 GCCGTGTGCG ACCCGGGCGC GAAACTCGCC CGCCGCGTGC GCGAAGCAGG 351 GTTCAAAGTC GTTCCCGTCG TGGGCGCAAG CGCGGTAATG GCGGCGTTGA 401 GTGTGGCCGG TGTGGCGGAA TCCGATTTTT ATTTCAACGG TTTTGTACCG 451 CCGAAATCGG GCGAACGTAG GAAATTGTTT GCCAAATGGG TGCGGGCGGC 501 ATTTCCTGTC GTCATGTTTG AAACGCCGCA CCGAATCGGG GCAACGCTTG 551 CCGATATGGC GGAATTGTTC CCCGAACGCC GTCTGATGCT GGCGCGCGAA 601 ATCACGAAAA CGTTTGAAAC GTTCTTAAGC GGCACGGTTG GGGAAATTCA 651 GACGGCATTG GCGGCGGACG GCAACCAATC GCGCGGCGAG ATGGTGTTGG 701 TGCTTTATCC GGCGCAGGAT GAAAAACACG AAGGCTTGTC CGAGTCTGCG 751 CAAAATGCGA TGAAAATCCT TGCGGCCGAG CTGCCGACCA AGCAGGCGGC 801 GGAGCTTGCC GCCAAGATTA CAGGTGAGGG CAAAAAGGCT TTGTACGATT 851 TGGCACTGTC GTGGAAAAAC AAATGA This coπesponds to the amino acid sequence <SEQ ID 646; ORF147ng-l>: 1 MFQKHLQKAS DSWGGTLYV VATPIGNLAD ITLRALAVLQ KADIICAEDT 51 RVTAQLLSAY GIQGRLVSVR EHNERQMADK VIGFLSDGLV VAQVSDAGTP 101 AVCDPGAKLA RRVREAGFKV VPWGASAVM AALSVAGVAE SDFYFNGFVP 151 PKSGERRKLF AKWVRAAFPV VMFETPHRIG ATLADMAELF PERRLMLARE 201 ITKTFETFLS GTVGEIQTAL AADGNQSRGE MVLVLYPAQD EKHEGLSESA 251 QNAMKILAAE LPTKQAAELA AKITGEGKKA LYDLALSWKN K* ORF147ng shows homology to a hypothetical E.coli protein: spl P45528I YRAL_ECOLI HYPOTHETICAL 31.3 KD PROTEIN IN AGAI-MTR INTERGENIC REGION (F286) >gi I 606086 (U18997) ORF_f286 [Escherichia coli] >gi I 1789535 (AE000395) hypothetical 31.3 kD protein in agai-mtr intergenic region [Escherichia coli] Length = 286 Score = 218 bits (550), Expect = 3e-56 Identities = 128/284 (45%), Positives = 171/284 (60%), Gaps = 4/284 (1%) Query: 4 KHLQKASDSWGGTLYWATPIGNLADITLRALAVLQKADIICAEDTRVTAQLLΞAYGIQ 63 K Q A +S G LY+V TPIGNLADIT RAL VLQ D+I AEDTR T LL +GI Sbjct: 2 KQHQSADNSQ—GQLYIVPTPIGNLADITQRALEVLQAVDLIAAEDTRHTGLLLQHFGIN 59 Query: 64 GRLVSVREHNERQMADKVIGFLSDGLWAQVSDAGTPAVCDPGAKLARRVREAGFKWPV 123 RL ++ +HNE+Q A+ ++ L +G +A VSDAGTP + DPG L R REAG +WP+ Sbjct: 60 ARLFALHDHNEQQKAETLLAKLQEGQNIALVSDAGTPLINDPGYHLVRTCREAGIRWPL 119 Query: 124 VGASAVMAALSVAGVAESDFYFNGFVPPKSGERRKLFAKWVRAAFPWMFETPHRIGATL 183 G A + ALS AG+ F + GF+P KS RR ++ +E+ HR+ +L Sbjct: 120 PGPCAAITALSAAGLPSDRFCYEGFLPAKSKGRRDALKAIEAEPRTLIFYESTHRLLDSL 179 Query: 184 ADMAELFPERR-LMLAREITKTFETFLSGTVGEIQTALAADGNQSRGEMVLVLYPAQDEK 242 D+ + E R ++LARE+TKT+ET VGE+ + D N+ +GEMVL++ + Sbjct: 180 EDIVAVLGESRYWLARELTKTWETIHGAPVGELLAWVKEDENRRKGEMVLIV-EGHKAQ 238 Query: 243 HEGLSESAQNAMKILAAELPTKQAAELAAKITGEGKKALYDLAL 286 E L A + +L AELP K+AA LAA+I G K ALY AL Sbjct: 239 EEDLPADALRTLALLQAELPLKKAAALAAEIHGVKKNALYKYAL 282 Based on the computer analysis and the presence of a putative transmembrane domain in the gonococcal protem, it is predicted that these proteins from N. meningitidis and N.gonorrhoeae, and their epitopes, could be useful antigens for vaccines or diagnostics, or for raising antibodies. Example 77 The following partial DNA sequence was identified in N. meningitidis <SEQ ID 647> 1 ATGAAAACAA CCGACAAACG GACAACCGAA ACACACCGCA AAGCCCCGAA 51 AACCGGTCGC ATCCGCTTCT C.GCTGCTTA CTTAGCCATA TGCCTGTCGT 101 TCGGCATTCT TCCCCAAGCC TGGGCGGGAC ACACTTATTT CGGCATCAAC 151 TACCAATACT ATCGCGACTT TGCCGAAAAT AAAGGCAAGT TTGCAGTCGG 201 GGCGAAAGAT ATTGAGGTTT ACAACAAAAA AGGGGAGTTG GTCGGCAAAT 251 CAATGACAAA AGCCCCGATG ATTGATTTTT CTGTGGTGTC GCGTAACGGC 301 GTGGCGGcAT TGGTGGGCGt ATCAATATAT TGTGAGCGTG GCACATAACG 351 GCGGCTATAA CAACGTTGAT TTTGGTGCGG AAGGAAk.AA tATCCC.GAT 401 CAACAwCGww TTACTTATAA AATTGTGAAA CGGAATAATT ATAAAGCAGG 451 GACTAAAGGC CATCCTTATG GCGGCGATTA TCATATGCCG CGTTTGCATA 501 AATwTGTCAC AGATGCAGAA CCTGTTGAAA TGACCAGTTA TATGGATGGG 551 CGGAAATATA TCGATCAAAA TAATTACCCT GACCGTGTTC GTATTGGGGC 601 AGGCAGGCAA TATTGGCGAT CTGATGAAGA TGAGCCCAAT AACCGCGAAA 651 GTTCATATCA TATTGCAAGT 701 GGCTC ACCAATGTTT ATCTATGATG CCCAAAAGCA 751 AAAGTGGTTA ATTAATGGGG TATTGCAAAC GGGCAACCCC TATATAGGAA 801 AAAGCAATGG CTTCCAGCTG GTTCGTAAAG ATTGGTTCTA TGATGAAATC 851 TTTGCTGGAG ATACCCATTC AGTATTCTAC GAACCACGTC AAAATGGGAA 901 ATACTCTTTT AACGACGATA ATAATGGCAC AGGAAAAATC AATGCCAAAC 951 ATGAACACAA TTCTCTGCCT AATAGATTAA AAACACGAAC CGTTCAATTG 1001 TTTAATGTTT CTTTATCCGA GACAGCAAGA GAACCTGTTT ATCATGCTGC 1051 AGGTGGTGTC AACAGTTATC GACCCAGACT GAATAATGGA GAAAATATTT 1101 CCTTTATTGA CGAAGGAAAA GGCGAATTGA TACTTACCAG CAACATCAAT 1151 CAAGGTGCTG GAGGATTATA TTTCCAAGGA GATTTTACGG TCTCGCCTGA 1201 AAATAACGAA ACTTGGCAAG GCGCGGGCGT TCATATCAGT GAAGACAGTA 1251 CCGTTACTTG GAAAGTAAAC GGCGTGGCAA ACGACCGCCT GTCCAAAATC 1301 GGCAAAGGCA CGCTG // 2101 GATAAAG 2151 TGACTGCTTC ATTGACTAAG ACCGACATCA GCGGCAATGT CGATCTTGCC 2201 GATCACGCTC ATTTAAATCT CACAGGGCTT GCCACACTCA ACGGCAATCT 2251 TAGTGCAAAT GGCGATACAC GTTATACAGT CAGCCACAAC GCCACCCAAA 2301 ACGGCAACCk TAgCCtCGtG G . sAATGcCC AAGCAACATT TAATCAAGCC 2351 ACATTAAACG GCAACACATC GGCTTCgGGC AATGCTTCAT TTAATCTAAG 2401 CGACCACGCC GTACAAAACG GCAGTCTGAC GCTTTCCGGC AACGCTAAGG 2451 CAAACGTAAG CCATTCCGCA CTCAACGGTA ATGTCTCCCT AGCCGATAAG 2501 GCAGTATTCC ATTTTGAAAG CAGCCGCTTT ACCGGACAAA TCAGCGGCGG 2551 CAagGATACG GCATTACACT TAAAAGACAG CGAATGGACG CTGCCGTCAg 2601 GarCGGAATT AGGCAATTTA AACCTTGACA ACGCCACCAT TACaCTCAAT 2651 TCCGCCTATC GCCACGATGC GGCAGGGGCG CAAACCGGCA GTGCGACAGA 2701 TGCGCCGCGC CGCCGTTCGC GCCGTTCGCG CCGTTCCCTA TTATmCGTTA 2751 CACCGCCAAC TTCGGTAGAA TCCCGTTTCA ACACGCTGAC GGTAAACGGC 2801 AAATTGAACG GTCAGGGAAC ATTCCGCTTT ATGTCGGAAC TCTTCGGCTA 2851 CCGCAGCGAC AAATTGAAGC TGGCGGAAAG TTCCGAAGGC ACTTACACCT 2901 TGGCGGTCAA CAATACCGGC AACGAACCTG CAAGCCTCGA ACAATTGACG 2951 GTAGTGGAAG GAAAAGACAA CAAACCGCTG TCCGAAAACC TTAATTTCAC 3001 CCTGCAAAAC GAACACGTCG ATGCAGGCGC GTGG // 3551 TTAGAC CGCGTATTTG CCGAAGACCG 3601 CCGCAACGCC GTTTGGACAA GCGGCATCCG GGACACCAAA CACTACCGTT 3651 CGCAAGATTT CCGCGCCTAC CGCCAACAAA CCGACCTGCG CCAAATCGGT 3701 ATGCAGAAAA ACCTCGGCAG CGGGCGCGTC GGCATCCTGT TTTCGCACAA 3751 CCGGACCGAA AACACCTTCG ACGACGGCAT CGGCAACTCG GCACGGCTTG 3801 CCCACGGCGC CGTTTTCGGG CAATACGGCA TCGACAGGTT CTACATCGGC 3851 ATCAGnCGCG GGCGCGGGTT TTAGCAGCGG CAGCCTTTcA GACGGCATCG 3901 GAGsmAAAwT CCGCCGCCGC GTGCtGCATT ACGGCATTCA GGCACGAtAC 3951 CGCGCCGgtt tCggCGgATt CGGCATCGAA CCGCACATCG GCGCAACGCg 4001 ctATTTCGTC CAAAAAGCGG ATTACCGCTA CGAAAACGTC AATATCGCCA 4051 CCCCCGGCCT TGCATTCAAC CGcTACCGCG CGGGCATTAa GGCAGATTAT 4101 TCATTCAAAC CGGCGCAACA CATTTCCATC ACGCCTTATT TGAGCCTGTC 4151 CTATACCGAT GCCGCTTCGG GCAAAGTCCG AACACGCGTC AATACCGCCG 4201 TATTGGCTCA GGATTTCGGC AAAACCCGCA GTGCGGAATG GGgCGTAAAC 4251 GCCGAAATCA AAGGTTTCAC GCTGTCCCTC CACGCTGCCG CCGCCAAAGG 4301 CCCGCAACTG GAAGCGCAAC ACAGCGCGGG CATCAAATTA GGCTACCGCT 4351 GGTAA... This coπesponds to the amino acid sequence <SEQ ID 648; ORFl>: 1 MKTTDKRTTE THRKAPKTGR IRFXAAYLAI CLSFGILPQA WAGHTYFGIN 51 YQYYRDFAEN KGKFAVGAKD IEVYNKKGEL VGKSMTKAPM IDFSWSRNG 101 VAALVGVQYI VSVAHNGGYN NVDFGAEGXN IXDQXRXTYK IVKRNNYKAG 151 TKGHPYGGDY HMPRLHKXVT DAEPVEMTSY MDGRKYIDQN NYPDRVRIGA 201 GRQYWRSDED EPNNRESSYH IAS GS PMFIYDAQKQ 251 KWLINGVLQT GNPYIGKSNG FQLVRKDWFY DEIFAGDTHS VFYEPRQNGK 301 YSFNDDNNGT GKINAKHEHN SLPNRLKTRT VQLFNVSLSE TAREPVYHAA 351 GGVNSYRPRL NNGENISFID EGKGELILTS NINQGAGGLY FQGDFTVSPE 401 NNETWQGAGV HISEDSTVTW KVNGVANDRL SKIGKGTL.. // 701 .... DKVTAS LTKTDISGNV DLADHAHLNL TGLATLNGNL 751 SANGDTRYTV SHNATQNGNX SLVXNAQATF NQATLNGNTS ASGNASFNLS 801 DHAVQNGSLT LSGNAKANVS HSALNGNVSL ADKAVFHFES SRFTGQISGG 851 KDTALHLKDS EWTLPSGXEL GNLNLDNATI TLNSAYRHDA AGAQTGSATD 901 APRRRSRRSR RSLLXVTPPT SVESRFNTLT VNGKLNGQGT FRFMSELFGY 951 RSDKLKLAES SEGTYTLAVN NTGNEPASLE QLTWEGKDN KPLSENLNFT 1001 LQNEHVDAGA W // 1151 LDRVFAEDR 1201 RNAVWTSGIR DTKHYRSQDF RAYRQQTDLR QIGMQKNLGS GRVGILFSHN 1251 RTENTFDDGI GNSARLAHGA VFGQYGIDRF YIGISAGAGF SSGSLSDGIG 1301 XKXRRRVLHY GIQARYRAGF GGFGIEPHIG ATRYFVQKAD YRYENV IAT 1351 PGLAFNRYRA GIKADYSFKP AQHISITPYL SLSYTDAASG KVRTRVNTAV 1401 LAQDFGKTRS AEWGVNAEIK GFTLSLHAAA AKGPQLEAQH SAGIKLGYRW 1451 * Further sequencing .analysis revealed the complete nucleotide sequence <SEQ ID 649>: 1 ATGAAAACAA CCGACAAACG GACAACCGAA ACACACCGCA AAGCCCCGAA 51 AACCGGCCGC ATCCGCTTCT CGCCTGCTTA CTTAGCCATA TGCCTGTCGT 101 TCGGCATTCT TCCCCAAGCC TGGGCGGGAC ACACTTATTT CGGCATCAAC 151 TACCAATACT ATCGCGACTT TGCCGAAAAT AAAGGCAAGT TTGCAGTCGG 201 GGCGAAAGAT ATTGAGGTTT ACAACAAAAA AGGGGAGTTG GTCGGCAAAT 251 CAATGACAAA AGCCCCGATG ATTGATTTTT CTGTGGTGTC GCGTAACGGC 301 GTGGCGGCAT TGGTGGGCGA TCAATATATT GTGAGCGTGG CACATAACGG 351 CGGCTATAAC AACGTTGATT TTGGTGCGGA AGGAAGAAAT CCCGATCAAC 401 ATCGTTTTAC TTATAAAATT GTGAAACGGA ATAATTATAA AGCAGGGACT 451 AAAGGCCATC CTTATGGCGG CGATTATCAT ATGCCGCGTT TGCATAAATT 501 TGTCACAGAT GCAGAACCTG TTGAAATGAC CAGTTATATG GATGGGCGGA 551 AATATATCGA TCAAAATAAT TACCCTGACC GTGTTCGTAT TGGGGCAGGC 601 AGGCAATATT GGCGATCTGA TGAAGATGAG CCCAATAACC GCGAAAGTTC 651 ATATCATATT GCAAGTGCGT ATTCTTGGCT CGTTGGTGGC AATACCTTTG 701 CACAAAATGG ATCAGGTGGT GGCACAGTCA ACTTAGGTAG TGAAAAAATT 751 AAACATAGCC CATATGGTTT TTTACCAACA GGAGGCTCAT TTGGCGACAG 801 TGGCTCACCA ATGTTTATCT ATGATGCCCA AAAGCAAAAG TGGTTAATTA 851 ATGGGGTATT GCAAACGGGC AACCCCTATA TAGGAAAAAG CAATGGCTTC 901 CAGCTGGTTC GTAAAGATTG GTTCTATGAT GAAATCTTTG CTGGAGATAC 951 CCATTCAGTA TTCTACGAAC CACGTCAAAA TGGGAAATAC TCTTTTAACG 1001 ACGATAATAA TGGCACAGGA AAAATCAATG CCAAACATGA ACACAATTCT 1051 CTGCCTAATA GATTAAAAAC ACGAACCGTT CAATTGTTTA ATGTTTCTTT 1101 ATCCGAGACA GCAAGAGAAC CTGTTTATCA TGCTGCAGGT GGTGTCAACA 1151 GTTATCGACC CAGACTGAAT AATGGAGAAA ATATTTCCTT TATTGACGAA 1201 GGAAAAGGCG AATTGATACT TACCAGCAAC ATCAATCAAG GTGCTGGAGG 1251 ATTATATTTC CAAGGAGATT TTACGGTCTC GCCTGAAAAT AACGAAACTT 1301 GGCAAGGCGC GGGCGTTCAT ATCAGTGAAG ACAGTACCGT TACTTGGAAA 1351 GTAAACGGCG TGGCAAACGA CCGCCTGTCC AAAATCGGCA AAGGCACGCT 1401 GCACGTTCAA GCCAAAGGGG AAAACCAAGG CTCGATCAGC GTGGGCGACG 1451 GTACAGTCAT TTTGGATCAG CAGGCAGACG ATAAAGGCAA AAAACAAGCC 1501 TTTAGTGAAA TCGGCTTGGT CAGCGGCAGG GGTACGGTGC AACTGAATGC 1551 CGATAATCAG TTCAACCCCG ACAAACTCTA TTTCGGCTTT CGCGGCGGAC 1601 GTTTGGATTT AAACGGGCAT TCGCTTTCGT TCCACCGTAT TCAAAATACC 1651 GATGAAGGGG CGATGATTGT CAACCACAAT CAAGACAAAG AATCCACCGT 1701 TACCATTACA GGCAATAAAG ATATTGCTAC AACCGGCAAT AACAACAGCT 1751 TGGATAGCAA AAAAGAAATT GCCTACAACG GTTGGTTTGG CGAGAAAGAT 1801 ACGACCAAAA CGAACGGGCG GCTCAACCTT GTTTACCAGC CCGCCGCAGA 1851 AGACCGCACC CTGCTGCTTT CCGGCGGAAC AAATTTAAAC GGCAACATCA 1901 CGCAAACAAA CGGCAAACTG TTTTTCAGCG GCAGACCAAC ACCGCACGCC 1951 TACAATCATT TAAACGACCA TTGGTCGCAA AAAGAGGGCA TTCCTCGCGG 2001 GGAAATCGTG TGGGACAACG ACTGGATCAA CCGCACATTT AAAGCGGAAA 2051 ACTTCCAAAT TAAAGGCGGA CAGGCGGTGG TTTCCCGCAA TGTTGCCAAA 2101 GTGAAAGGCG ATTGGCATTT GAGCAATCAC GCCCAAGCAG TTTTTGGTGT 2151 CGCACCGCAT CAAAGCCACA CAATCTGTAC ACGTTCGGAC TGGACGGGTC 2201 TGACAAATTG TGTCGAAAAA ACCATTACCG ACGATAAAGT GATTGCTTCA 2251 TTGACTAAGA CCGACATCAG CGGCAATGTC GATCTTGCCG ATCACGCTCA 2301 TTTAAATCTC ACAGGGCTTG CCACACTCAA CGGCAATCTT AGTGCAAATG 2351 GCGATACACG TTATACAGTC AGCCACAACG CCACCCAAAA CGGCAACCTT 2401 AGCCTCGTGG GCAATGCCCA AGCAACATTT AATCAAGCCA CATTAAACGG 2451 CAACACATCG GCTTCGGGCA ATGCTTCATT TAATCTAAGC GACCACGCCG 2501 TACAAAACGG CAGTCTGACG CTTTCCGGCA ACGCTAAGGC AAACGTAAGC 2551 CATTCCGCAC TCAACGGTAA TGTCTCCCTA GCCGATAAGG CAGTATTCCA 2601 TTTTGAAAGC AGCCGCTTTA CCGGACAAAT CAGCGGCGGC AAGGATACGG 2651 CATTACACTT AAAAGACAGC GAATGGACGC TGCCGTCAGG CACGGAATTA 2701 GGCAATTTAA ACCTTGACAA CGCCACCATT ACACTCAATT CCGCCTATCG 2751 CCACGATGCG GCAGGGGCGC AAACCGGCAG TGCGACAGAT GCGCCGCGCC 2801 GCCGTTCGCG CCGTTCGCGC CGTTCCCTAT TATCCGTTAC ACCGCCAACT 2851 TCGGTAGAAT CCCGTTTCAA CACGCTGACG GTAAACGGCA AATTGAACGG 2901 TCAGGGAACA TTCCGCTTTA TGTCGGAACT CTTCGGCTAC CGCAGCGACA 2951 AATTGAAGCT GGCGGAAAGT TCCGAAGGCA CTTACACCTT GGCGGTCAAC 3001 AATACCGGCA ACGAACCTGC AAGCCTCGAA CAATTGACGG TAGTGGAAGG 3051 AAAAGACAAC AAACCGCTGT CCGAAAACCT TAATTTCACC CTGCAAAACG 3101 AACACGTCGA TGCCGGCGCG TGGCGTTACC AACTCATCCG CAAAGACGGC 3151 GAGTTCCGCC TGCATAATCC GGTCAAAGAA CAAGAGCTTT CCGACAAACT 3201 CGGCAAGGCA GAAGCCAAAA AACAGGCGGA AAAAGACAAC GCGCAAAGCC 3251 TTGACGCGCT GATTGCGGCC GGGCGCGATG CCGTCGAAAA GACAGAAAGC 3301 GTTGCCGAAC CGGCCCGGCA GGCAGGCGGG GAAAATGTCG GCATTATGCA 3351 GGCGGAGGAA GAGAAAAAAC GGGTGCAGGC GGATAAAGAC ACCGCCTTGG 3401 CGAAACAGCG CGAAGCGGAA ACCCGGCCGG CTACCACCGC CTTCCCCCGC 3451 GCCCGCCGCG CCCGCCGGGA TTTGCCGCAA CTGCAACCCC AACCGCAGCC 3501 CCAACCGCAG CGCGACCTGA TCAGCCGTTA TGCCAATAGC GGTTTGAGTG 3551 AATTTTCCGC CACGCTCAAC AGCGTTTTCG CCGTACAGGA CGAATTAGAC 3601 CGCGTATTTG CCGAAGACCG CCGCAACGCC GTTTGGACAA GCGGCATCCG 3651 GGACACCAAA CACTACCGTT CGCAAGATTT CCGCGCCTAC CGCCAACAAA 3701 CCGACCTGCG CCAAATCGGT ATGCAGAAAA ACCTCGGCAG CGGGCGCGTC 3751 GGCATCCTGT TTTCGCACAA CCGGACCGAA AACACCTTCG ACGACGGCAT 33880011 CGGCAACTCG GCACGGCTTG CCCACGGCGC CGTTTTCGGG CAATACGGCA 3851 TCGACAGGTT CTACATCGGC ATCAGCGCGG GCGCGGGTTT TAGCAGCGGC 3901 AGCCTTTCAG ACGGCATCGG AGGCAAAATC CGCCGCCGCG TGCTGCATTA 3951 CGGCATTCAG GCACGATACC GCGCCGGTTT CGGCGGATTC GGCATCGAAC 4001 CGCACATCGG CGCAACGCGC TATTTCGTCC AAAAAGCGGA TTACCGCTAC 44005511 GAAAACGTCA ATATCGCCAC CCCCGGCCTT GCATTCAACC GCTACCGCGC 4101 GGGCATTAAG GCAGATTATT CATTCAAACC GGCGCAACAC ATTTCCATCA 4151 CGCCTTATTT GAGCCTGTCC TATACCGATG CCGCTTCGGG CAAAGTCCGA 4201 ACACGCGTCA ATACCGCCGT ATTGGCTCAG GATTTCGGCA AAACCCGCAG 4251 TGCGGAATGG GGCGTAAACG CCGAAATCAA AGGTTTCACG CTGTCCCTCC 44330011 ACGCTGCCGC CGCCAAAGGC CCGCAACTGG AAGCGCAACA CAGCGCGGGC 4351 ATCAAATTAG GCTACCGCTG GTAA This coπesponds to the amino acid sequence <SEQ ID 650; ORFl-l>: 1 MKTTDKRTTE THRKAPKTGR IRFSPAYLAI CLSFGILPQA WAGHTYFGIN 51 YQYYRDFAEN KGKFAVGAKD IEVYNKKGEL VGKSMTKAPM IDFSWSRNG 101 VAALVGDQYI VSVAHNGGYN NVDFGAEGRN PDQHRFTYKI VKRNNYKAGT 151 KGHPYGGDYH MPRLHKFVTD AEPVEMTSYM DGRKYIDQNN YPDRVRIGAG 201 RQYWRSDEDE PNNRESSYHI ASAYSWLVGG NTFAQNGSGG GTVNLGSEKI 251 KHSPYGFLPT GGSFGDSGSP MFIYDAQKQK WLINGVLQTG NPYIGKSNGF 301 QLVRKDWFYD EIFAGDTHSV FYEPRQNGKY SFNDDNNGTG KINAKHEHNS 351 LPNRLKTRTV QLFNVSLSET AREPVYHAAG GVNSYRPRLN NGENISFIDE 401 GKGELILTSN INQGAGGLYF QGDFTVSPEN NETWQGAGVH ISEDSTVTWK 451 VNGVANDRLS KIGKGTLHVQ AKGENQGSIS VGDGTVILDQ QADDKGKKQA 501 FSEIGLVSGR GTVQLNADNQ FNPDKLYFGF RGGRLDLNGH SLSFHRIQNT 551 DEGAMIVNHN QDKESTVTIT GNKDIATTGN NNSLDSKKEI AYNGWFGEKD 601 TTKTNGRLNL VYQPAAEDRT LLLSGGTNLN GNITQTNGKL FFSGRPTPHA 651 YNHLNDHWSQ KEGIPRGEIV WDNDWINRTF KAENFQIKGG QAWSRNVAK 701 VKGDWHLSNH AQAVFGVAPH QSHTICTRSD WTGLTNCVEK TITDDKVIAS 751 LTKTDISGNV DLADHAHLNL TGLATLNGNL SANGDTRYTV SHNATQNGNL 801 SLVGNAQATF NQATLNGNTS ASGNASFNLS DHAVQNGSLT LSGNAKANVS 851 HSALNGNVSL ADKAVFHFES SRFTGQISGG KDTALHLKDS EWTLPSGTEL 901 GNLNLDNATI TLNSAYRHDA AGAQTGSATD APRRRSRRSR RSLLSVTPPT 951 SVESRFNTLT VNGKLNGQGT FRFMSELFGY RSDKLKLAES SEGTYTLAVN 1001 NTGNEPASLE QLTWEGKDN KPLSENLNFT LQNEHVDAGA WRYQLIRKDG 1051 EFRLHNPVKE QELSDKLGKA EAKKQAEKDN AQSLDALIAA GRDAVEKTES 1101 VAEPARQAGG ENVGIMQAEE EKKRVQADKD TALAKQREAE TRPATTAFPR 1151 ARRARRDLPQ LQPQPQPQPQ RDLISRYANS GLSEFSATLN SVFAVQDELD 1201 RVFAEDRRNA VWTSGIRDTK HYRSQDFRAY RQQTDLRQIG MQKNLGSGRV 1251 GILFSHNRTE NTFDDGIGNS ARLAHGAVFG QYGIDRFYIG ISAGAGFSSG 1301 SLSDGIGGKI RRRVLHYGIQ ARYRAGFGGF GIEPHIGATR YFVQKADYRY 1351 ENVNIATPGL AFNRYRAGIK ADYSFKPAQH ISITPYLSLS YTDAASGKVR 1401 TRVNTAVLAQ DFGKTRSAEW GVNAEIKGFT LSLHAAAAKG PQLEAQHSAG 1451 IKLGYRW* Computer analysis of these sequences gave the following results: Homology with a predicted ORF from N.meninsitidis (strain Al ORFl shows 57.8% identity over a 1456aa overlap with an ORF (ORFla) from sfrain A of N. meningitidis: 10 20 30 40 50 60 orf1. pep MKTTDKRTTETHRKAPKTGRIRFXAAYLAICLSFGILPQAWAGHTYFGINYQYYRDFAEN I I I I I II I II I I I II I II I I II I I I I I I I I I M I I I I I II I I I II II I I II II I I II I orfla MKTTDKRTTETHRKAPKTGRIRFSPAYLAICLSFGILPQAWAGHTYFGINYQYYRDFAEN 10 20 30 40 50 60 70 80 90 100 110 120 orf1. pep KGKFAVGAKDIEVYNKKGELVGKSMTKAPMIDFSWSRNGVAALVGVQYIVSVAHNGGYN II I II II II I I I II I II II II I II II I II II II I II I II II I I II I I I I II I I II I I II orfla KGKFAVGAKDIEVYNKKGELVGKSMTKAPMIDFSWSRNGVAALVGDQYIVSVAHNGGYN 70 80 90 100 110 120 130 140 150 160 170 180 orf1. pep NVDFGAEGXNIXDQXRXTYKIVKRNNYKAGTKGHPYGGDYHMPRLHKXVTDAEPVEMTSY I II II II II I II I : I : II I I I I II : : M M II II I I I I I I I I II I II I orfla NVDFGAEGXN-PDQHRFSYQIVKRNNYKPDNS-HPYNGDXHMPRLHKFVTDAEPVEMTSD 130 140 150 160 170 190 200 210 orf1. pep MDGRKYIDQNNYPDRVRIGAGRQYWRSDEDEP NN I I I I ::: I I: I I I I I: I:: M I I : I : II orfla MRGNTYSDKEKYPERVRIGSGHHYWRYDDDKHGDLSYSGAWLIGGNTHMQGWGNNGVXSL 180 190 200 210 220 230 220 230 240 250 260 orf1. pep RESSYH IA SGSPMFIYDAQKQKWLINGVLQTGNPYIGKSNGFQLVRK I : : : : II I II I II I I I : M I I : I I I I I I I II I : I II I I : II orfla SGDVRHANDYGPMPIAGAAGDSGSPMFIYDKTNNKWLLNGVLQTGYPYSGRENGFQLIRK 240 250 260 270 280 290 270 280 290 300 310 320 orf1. pep DWFYDEIFAGDTHSVFYEPRQNGKYSFNDDNNGTGKINAKHEHNSLPNRLKTRTVQLFNV I M II : I : I II I : I : I I I : I I : : I I : : : I II I I : : : I : I I : II : : II : II : orfla DWFYDDIYRGDTHTVXFEPRSNGHFSFTSNNNGTGTVTETNEKVSNP-KLKVQTVRLFDE 300 310 320 330 340 350 330 340 350 360 370 380 orf1.pep SLSETAREPVYHAAGGVNSYRPRLNNGENISFIDEGKGELILTSNINQGAGGLYFQGDFT IMM Mill I I I I I I : I I II I II I I I : I I II I : I : I I I : : I I I I I I I I I I I : I I I I orfla SLNETDKEPVY-AAGGVNQYRPRLNNGENLSFIDYGNGKLILSNNINQGAGGLYFEGDFT 360 370 380 390 400 410 390 400 410 420 430 orf1. pep VSPENNETWQGAGVHISEDSTVTWKVNGVANDRLSKIGKGTL II I II II I I I I I I I I II II I II I I I I II II I II I II I II II I orfla VSPENNETWQGAGVHISEDSTVTWKVNGVANDRLSKIGKGTLHVQAKGENQGSISVGDGT 420 430 440 450 460 470 orf1.pep orfla VILDQQADDKGKKQAFSEIGLXSGRGTVQLNADNQFNPDKLYFGFRGGRLDLNGHSLSFH 480 490 500 510 520 530 orf1.pep orfla RIQNTDEGAMIXXHNATTTSTVTITGNEΞITQPSGKNINRLNYSKEIAYNGWFGEKDTTK 540 550 560 570 580 590 orf1.pep orfla TNGRLNLVYQPAAEDRTXLLSGGTNLNGNITQTNGKLFFSGRPTPHAYNHLGSGWSKMEG 600 610 620 630 640 650 orf1.pep orfla IPQGEIVWDNDWIXRTFKAENFHIQGGQAVISRNVAKVEGDXHLSNHAQAVFGVAPHQSH 660 670 680 690 700 710 440 450 460 470 480 orf1. pep XXXXXDKVTASLTKTDISGNVDLADHAHLNLTGLATLNGNLSAN : 11 : 111 1111111 11 1 1 IM I II II II orfla TICTRSDWTGLTNCVEXXITDDKVIASLTKTDXSGXVXLXXXXXXXLXGXAXLXGNLSAN 720 730 740 750 760 770 490 500 510 520 530 540 orf1. pep GDTRYTVSHNAT Q NGNXSLVXNAQATFNQATLNGNTSASGNASFNLSDHAVQNGSLTLSG II I II I I I II I I II I I III II I II I I II II II I: I I II I I I I I I :: I : I I I II II I orfla GDTRYTVSHNATQNGNLSLVGNAQATFNQATLNGNXSXSGNASFNLSNNAAQNGSLTLSD 780 790 800 810 820 830 550 560 570 580 590 600 orf1. pep NAKANVSHSALNGNVSLADKAVFHFESSRFTGQISGGKDTALHLKDSEWTLPSGXELGNL I I I I I II II I II II I I II I I I II I I I: I II II I : I I : I II I I I II I I I I II I I: I I I I I orfla NAKANVSHSALNGNVSLADKAVFHFENSRFTGQLSGSKXTALHLKDSEWTLPSGTELGNL 840 850 860 870 880 890 610 620 630 640 650 660 orf1. pep NLDNATITLNSAYRHDAAGAQTGSATDAPRRRSRRSRRSLLXVTPPTSVESRFNTLTVNG II I I II II I II II I I I I II I II I :: I : I I I I I I I I II I I I I I I I I II I I I I I I I I orfla NLDNATITLNSAYRHDAAGAQTGXVSDTPRRRSRRS LLSVTPPTSVESRFNTLTVNG 900 910 920 930 940 950 670 680 690 700 710 720 orf1. pep KLNGQGTFRFMSELFGYRSDKLKLAESSEGTYTLAVNNTGNEPASLEQLTWEGKDNKPL III I I I I I I II II I I I I II I I I II I I I I I I I I I I I I I I II I I : II : I I II I I I I I I I I I orfla KLNXQGTFRFMSELFGYRSDKLKLAESSEGTYTLAVNNTGNEPVSLDQLTWEGKDNKPL 960 970 980 990 1000 1010 730 740 750 orf1. pep SENLNFTLQNEHVDAGAW I I I II I I I II I II I II II orfla SENLNFTLQNEHVDAGAWRYQLIRKDGEFRLHNPVKEQELSDKLGKAEAKKQAEKDNAQS 1020 1030 1040 1050 1060 1070 orf1.pep orfla LDALIAAGRDAAEKTESVAEPARXAGGENVGIMQAEEEKKRVQADKDSALAKQREAETRP 1080 1090 1100 1110 1120 1130 760 orf1. pep LDR I I I orfla XTTAFPRARXARRDLPQPQPQPQPQPQPQRDLXSRYANSGLSEFSATLNSVFAVQDELDR 1140 1150 1160 1170 1180 1190 770 780 790 800 810 820 orf1. pep VFAEDRRNAVWTSGIRDTKHYRSQDFRAYRQQTDLRQIGMQKNLGSGRVGILFSHNRTEN I I II I II I II I II II I II I I I I I I I I I I I I II I II II I II I I I I I I I I I I II I I I I I I orfla VFAEDRRNAVWTSXIRXTKHYRSQDFRAYRQQTDLRQIGMQKNLGSGRVGILFSHNRTEN 1200 1210 1220 1230 1240 1250 830 840 850 860 870 880 orf1. pep TFDDGIGNSARLAHGAVFGQYGIDRFYIGISAGAGFSSGSLSDGIGXKXRRRVLHYGIQA : II I I I I I I I II I II I II I I I I I II I I I I : I I I I I II I I II I I I I II I I I I I I I I orfla XFDDGIGNSARLAHGAVFGQYGIGRFDIGISTGAGFSSGXLSDGIGGKIRRRVLHYGIQA 1260 1270 1280 1290 1300 1310 890 900 910 920 930 940 orf1. pep RYRAGFGGFGIEPHIGATRYFVQKADYRYENVNIATPGLAFNRYRAGIKADYSFKPAQHI II II I I I I I I II I: I I II II I I I II I II II II I I II II I I I I I I II I II I I II II I I II orfla RYRAGFGGFGIEPYIGATRYFVQKADYRYENVNIATPGLAFNRYRAGIKADYSFKPAQHX 1320 1330 1340 1350 1360 1370 950 960 970 980 990 1000 orf1. pep SITPYLSLSYTDAASGKVRTRVNTAVLAQDFGKTRSAEWGVNAEIKGFTLSLHAAAAKGP I I I I I I I I I II I II I II II I I I II I II II I I I II I II II I I I I I I I I I I I I I I II I II orfla SITPYXSLSYTDAASGKVRTRVNTAVLAQDFGKTRSAEWGVNAEIKGFTLSXHAAAAKGP 1380 1390 1400 1410 1420 1430 1010 1020 orf1. pep QLEAQHSAGIKLGYRWX I I I II I II I II I II I I I orfla QLEAQHSAGIKLGYRWX 1440 1450 The complete length ORF 1 a nucleotide sequence <SEQ LD 651 > is: 1 ATGAAAACAA CCGACAAACG GACAACCGAA ACACACCGCA AAGCCCCGAA 51 AACCGGCCGC ATCCGCTTCT CGCCTGCTTA CTTAGCCATA TGCCTGTCGT 101 TCGGCATTCT TCCCCAAGCT TGGGCGGGAC ACACTTATTT CGGCATCAAC 151 TACCAATACT ATCGCGACTT TGCCGAAAAT AAAGGCAAGT TTGCAGTCGG 201 GGCGAAAGAT ATTGAGGTNT ACAACAAAAA AGGGGAGTTG GTCGGCAAAT 251 CAATGACAAA AGCCCCGATG ATTGATTTTT CTGTGGTGTC GCGTAACGGC 301 GTGGCGGCAT TGGTGGGCGA TCAATATATT GTGAGCGTGG CACATAACGG 351 CGGCTATAAC AACGTTGATT TTGGTGCGGA AGGAAGNAAT CCCGATCAGC 401 ACCGTTTTTC TTACCAAATT GTGAAAAGAA ATAATTATAA GCCTGACAAT 451 TCACACCCTT ACAACGGCGA TTANCATATG CCGCGTTTGC ATAAATTTGT 501 CACAGATGCA GAACCTGTCG AAATGACGAG TGACATGAGG GGGAATACCT 551 ATTCCGATAA AGAAAAATAT CCCGAGCGTG TCCGCATCGG CTCAGGACAC 601 CACTATTGGC GTTATGATGA TGACAAACAC GGCGATTTAT CCTACTCCGG 651 CGCATGGTTA ATTGGCGGCA ATACACATAT GCAGGGTTGG GGAAATAATG 701 GCGTANTTAG TTTGAGCGGC GATGTGCGCC ATGCCAACGA CTATGGCCCT 751 ATGCCGATTG CAGGTGCGGC AGGCGACAGC GGTTCGCCAA TGTTTATTTA 801 TGACAAAACA AACAATAAAT GGCTGCTCAA CGGAGTTTTA CAAACCGGCT 851 ACCCTTATTC CGGCAGGGAA AACGGTTTCC AGCTGATACG CAAAGATTGG 901 TTCTACGATG ACATTTACAG AGGCGATACA CATACCGTCT NTTTTGAACC 951 GCGCAGTAAC GGACATTTTT CCTTTACATC CAACAACAAC GGTACGGGTA 1001 CGGTAACAGA AACCAACGAA AAGGTNTCCA ATCCAAAGCT TAAAGTACAG 1051 ACAGTCCGAC TGTTTGACGA ATCTTTGAAT GAAACTGATA AAGAACCAGT 1101 TTACGCGGCA GGGGGTGTTA ATCAGTACCG TCCAAGGTTA AACAACGGTG 1151 AAAACCTTTC TTTTATCGAT TACGGCAACG GCAAACTCAT CTTATCAAAC 1201 AACATCAACC AAGGCGCGGG CGGTTTGTAT TTTGAAGGTG ATTTTACGGT 1251 CTCGCCTGAA AACAACGAAA CGTGGCAAGG CGCGGGCGTT CATATCAGTG 1301 AAGACAGTAC CGTTACTTGG AAAGTAAACG GCGTGGCAAA CGACCGCCTG 1351 TCCAAAATCG GCAAAGGCAC GCTGCACGTT CAAGCCAAAG GGGAAAACCA 1401 AGGCTCGATC AGCGTGGGCG ACGGTACAGT CATTTTGGAT CAGCAGGCAG 1451 ACGATAAAGG CAAAAAACAA GCCTTTAGTG AAATCGGCTT GNTCAGCGGC 1501 AGGGGTACGG TGCAACTGAA TGCCGATAAT CAGTTCAACC CCGACAAACT 1551 CTATTTCGGC TTTCGCGGCG GACGTTTGGA TTTAAACGGG CATTCGCTTT 1601 CGTTCCACCG TATTCAAAAT ACCGATGAAG GGGCGATGAT TGNCNATCAT 1651 AATGCCACAA CAACATCCAC CGTTACCATT ACAGGGAATG AAAGTATTAC 1701 ACAACCGAGT GGTAAGAATA TCAATAGACT TAATTACAGC AAAGAAATTG 1751 CCTACAACGG TTGGTTTGGC GAGAAAGATA CGACCAAAAC GAACGGGCGG 1801 CTCAACCTTG TTTACCAGCC CGCCGCAGAA GACCGCACCC NGCTGCTTTC 1851 CGGCGGAACA AATTTAAACG GCAACATCAC GCAAACAAAC GGCAAACTGT 1901 TTTTCAGCGG CAGACCGACA CCGCACGCCT ACAATCATTT AGGAAGCGGG 1951 TGGTCAAAAA TGGAAGGTAT CCCACAAGGA GAAATCGTGT GGGACAACGA 2001 CTGGATCNAC CGCACGTTTA AAGCGGAAAA TTTCCATATT CAGGGCGGGC 2051 AGGCGGTGAT TTCCCGCAAT GTTGCCAAAG TGGAAGGCGA TTGNCATTTG 2101 AGCAATCACG CCCAAGCAGT TTTTGGTGTC GCACCGCATC AAAGCCATAC 2151 AATCTGTACA CGTTCGGACT GGACNGGTCT GACAAATTGT GTCGAANAAA 2201 NCATTACCGA CGATAAAGTG ATTGCTTCAT TGACTAAGAC NGACNTNAGC 2251 GGCANTGTNA GNCTNNCCNA TNACGNTNNT TNAAANCTCN CNGGGCNTGC 2301 NNCACTNAAN GGCAATCTTA GTGCAAATGG CGATACACGT TATACAGTCA 2351 GCCACAACGC CACCCAAAAC GGCAACCTTA GCCTCGTGGG CAATGCCCAA 2401 GCAACATTTA ATCAAGCCAC ATTAAACGGC AACNCATCGG NTTCGGGCAA 2451 TGCTTCATTT AATCTAAGCA ACAACGCCGC ACAAAACGGC AGTCTGACGC 2501 TTTCCGACAA CGCTAAGGCA AACGTAAGCC ATTCCGCACT CAACGGCAAT 2551 GTCTCCCTAG CCGATAAGGC AGTATTCCAT TTTGAAAACA GCCGCTTTAC 2601 CGGACAACTC AGCGGCAGCA AGGANACAGC ATTACACTTA AAAGACAGCG 2651 AATGGACGCT GCCGTCAGGC ACGGAATTAG GCAATTTAAA CCTTGACAAC 2701 GCCACCATTA CACTCAATTC CGCCTATCGC CACGATGCTG CAGGCGCGCA 2751 AACCGGCAGN GTGTCAGACA CGCCGCGCCG CCGTTCGCGC CGTTCCCTAT 2801 TATCCGTTAC ACCGCCAACT TCGGTAGAAT CCCGTTTCAA CACGCTGACG 2851 GTAAACGGCA AATTGAACNG TCAAGGAACA TTCCGCTTTA TGTCGGAACT 2901 CTTCGGCTAC CGAAGCGACA AATTGAAGCT GGCGGAAAGT TCCGAAGGNA 2951 CTTACACCTT GGCGGTCAAC AATACCGGCA ACGAACCCGT AAGCCTCGAT 3001 CAATTGACGG TAGTGGAAGG GAAAGACAAC AAACCGCTGT CCGAAAACCT 3051 TAATTTCACC CTGCAAAACG AACACGTCGA TGCCGGCGCG TGGCGTTACC 3101 AACTCATCCG CAAAGACGGC GAGTTCCGCC TGCATAATCC GGTCAAAGAA 3151 CAAGAGCTTT CCGACAAACT CGGCAAGGCA GAAGCCAAAA AACAGGCGGA 3201 AAAAGACAAC GCGCAAAGCC TTGACGCGCT GATTGCGGCC GGGCGCGATG 3251 CCGCCGAAAA GACAGAAAGC GTTGCCGAAC CGGCCCGGCN GGCAGGCGGG 3301 GAAAATGTCG GCATTATGCA GGCGGAGGAA GAGAAAAAAC GGGTGCAGGC 3351 GGATAAAGAC AGCGCNTTGG CGAAACAGCG CGAAGCGGAA ACCCGGCCGG 3401 NTACCACCGC CTTCCCCCGC GCCCGCNGCG CCCGCCGGGA TTTGCCGCAA 3451 CCGCAGCCCC AACCGCAACC TCAACCCCAA CCGCAGCGCG ACCTGATNAG 3501 CCGTTATGCC AATAGCGGTT TGAGTGAATT TTCCGCCACG CTCAACAGCG 3551 TTTTCGCCGT ACAGGACGAA TTGGACCGCG TGTTTGCCGA AGACCGCCGC 3601 AACGCNGTTT GGACAAGCNG CATCCGGNAC ACCAAACACT ACCGTTCGCA 3651 AGATTTCCGC GCCTACCGCC AACAAACCGA CCTGCGCCAA ATCGGTATGC 3701 AGAAAAACCT CGGCAGCGGG CGCGTCGGCA TCCTGTTTTC GCACAACCGG 3751 ACCGAAAACA NCTTCGACGA CGGCATCGGC AACTCGGCAC GGCTTGCCCA 3801 CGGCGCCGTT TTCGGGCAAT ACGGCATCGG CAGGTTCGAC ATCGGCATCA 3851 GCACGGGCGC GGGTTTTAGC AGCGGCANTC TNTCAGACGG CATCGGAGGC 3901 AAAATCCGCC GCCGCGTGCT GCATTACGGC ATTCAGGCAC GATACCGCGC 3951 CGGTTTCGGC GGATTCGGCA TCGAACCGTA CATCGGCGCA ACGCGCTATT 4001 TCGTCCAAAA AGCGGATTAC CGCTACGAAA ACGTCAATAT CGCCACCCCC 4051 GGTCTTGCGT TCAACCGNTA CCGNGCGGGC ATTAAGGCAG ATTATTCATT 4101 CAAACCGGCG CAACACATNT CCATCACNCC TTATTTNAGC CTGTCCTATA 4151 CCGATGCCGC TTCGGGCAAA GTCCGAACAC GCGTCAATAC CGCNGTATTG 4201 GCTCAGGATT TCGGCAAAAC CCGCAGTGCG GAATGGGGCG TAAACGCCGA 4251 AATCAAAGGT TTCACGCTGT CCNTCCACGC TGCCGCCGCC AAAGGNCCGC 4301 AACTGGAAGC GCAACACAGC GCGGGCATCA AATTAGGCTA CCGCTGGTAA This encodes a protein having amino acid sequence <SEQ LD 652>: 1 MKTTDKRTTE THRKAPKTGR IRFSPAYLAI CLSFGILPQA WAGHTYFGIN 51 YQYYRDFAEN KGKFAVGAKD IEVYNKKGEL VGKSMTKAPM IDFSWSRNG 101 VAALVGDQYI VSVAHNGGYN NVDFGAEGXN PDQHRFSYQI VKRNNYKPDN 151 SHPYNGDXHM PRLHKFVTDA EPVEMTSDMR GNTYSDKEKY PERVRIGSGH 201 HYWRYDDDKH GDLSYSGAWL IGGNTHMQGW GNNGVXSLSG DVRHANDYGP 251 MPIAGAAGDS GSPMFIYDKT NNKWLLNGVL QTGYPYSGRE NGFQLIRKDW 301 FYDDIYRGDT HTVXFEPRSN GHFSFTSNNN GTGTVTETNE KVSNPKLKVQ 351 TVRLFDESLN ETDKEPVYAA GGVNQYRPRL NNGENLSFID YGNGKLILSN 401 NINQGAGGLY FEGDFTVSPE NNETWQGAGV HISEDSTVTW KVNGVANDRL 451 SKIGKGTLHV QAKGENQGSI SVGDGTVILD QQADDKGKKQ AFSEIGLXSG 501 RGTVQLNADN QFNPDKLYFG FRGGRLDLNG HSLSFHRIQN TDEGAMIXXH 551 NATTTSTVTI TGNESITQPS GKNINRLNYS KEIAYNGWFG EKDTTKTNGR 601 LNLVYQPAAE DRTXLLSGGT NLNGNITQTN GKLFFSGRPT PHAYNHLGSG 651 WSKMEGIPQG EIVWDNDWIX RTFKAENFHI QGGQAVISRN VAKVEGDXHL 701 SNHAQAVFGV APHQSHTICT RSDWTGLTNC VEXXITDDKV IASLTKTDXS 751 GXVXLXXXXX XXLXGXAXLX GNLSANGDTR YTVSHNATQN GNLSLVGNAQ 801 ATFNQATLNG NXSXSGNASF NLSNNAAQNG SLTLSDNAKA NVSHSALNGN 851 VSLADKAVFH FENSRFTGQL SGSKXTALHL KDSEWTLPSG TELGNLNLDN 901 ATITLNSAYR HDAAGAQTGX VSDTPRRRSR RSLLSVTPPT SVESRFNTLT 951 VNGKLNXQGT FRFMSELFGY RSDKLKLAES SEGTYTLAVN NTGNEPVSLD 1001 QLTWEGKDN KPLSENLNFT LQNEHVDAGA WRYQLIRKDG EFRLHNPVKE 1051 QELSDKLGKA EAKKQAEKDN AQSLDALIAA GRDAAEKTES VAEPARXAGG 1101 ENVGIMQAEE EKKRVQADKD SALAKQREAE TRPXTTAFPR ARXARRDLPQ 1151 PQPQPQPQPQ PQRDLXSRYA NSGLSEFSAT LNSVFAVQDE LDRVFAEDRR 1201 NAVWTSXIRX TKHYRSQDFR AYRQQTDLRQ IGMQKNLGSG RVGILFSHNR 1251 TENXFDDGIG NSARLAHGAV FGQYGIGRFD IGISTGAGFS SGXLSDGIGG 1301 KIRRRVLHYG IQARYRAGFG GFGIEPYIGA TRYFVQKADY RYENVNIATP 1351 GLAFNRYRAG IKADYSFKPA QHXSITPYXS LSYTDAASGK VRTRVNTAVL 1401 AQDFGKTRSA EWGVNAEIKG FTLSXHAAAA KGPQLEAQHS AGIKLGYRW* A transmembrane region is underlined. ORFl-1 shows 86.3% identity over a 1462aa overlap with ORFla: 10 20 30 40 50 60 orfla . pep MKTTDKRTTETHRKAPKTGRIRFSPAYLAICLSFGILPQAWAGHTYFGINYQYYRDFAEN I I II I I I I I I I I I I I I II I II I II I II I I II I I I I I I I I I I I I II I II I I I I II I I II I I orf1-1 MKTTDKRTTETHRKAPKTGRIRFSPAYLAICLSFGILPQAWAGHTYFGINYQYYRDFAEN 10 20 30 40 50 60 70 80 90 100 110 120 orfla . pep KGKFAVGAKDIEVYNKKGELVGKSMTKAPMIDFSWSRNGVAALVGDQYIVSVAHNGGYN I I II II I I I I II II II I II I I II I I I I I II I II II II I II I I I I I I I I I II I I I I II I II orf1-1 KGKFAVGAKDIEVYNKKGELVGKSMTKAPMIDFSWSRNGVAALVGDQYIVSVAHNGGYN 70 80 90 100 110 120 130 140 150 160 170 179 orfla . pep NVDFGAEGXNPDQHRFSYQIVKRNNYKPDNS-HPYNGDXHMPRLHKFVTDAEPVEMTSDM I I II I I I I I I I I I IM I: I I II I I I I :: M M I II I I II I II I I I II II I I I orf1-1 NVDFGAEGRNPDQHRFTYKIVKRNNYKAGTKGHPYGGDYHMPRLHKFVTDAEPVEMTSYM 130 140 150 160 170 180 180 190 200 210 220 230 orfla . pep RGNTYSDKEKYPERVRIGSGHHYWRYDDDKHGDL—SYSGA WLIGGNTHMQGWGNN I I I : :: I I : II II I : I :: M I I : I : : : II I I I : II II I : ::: orf1-1 DGRKYIDQNNYPDRVRIGAGRQYWRSDEDEPNNRESSYHIASAYSWLVGGNTFAQNGSGG 190 200 210 220 230 240 240 250 260 270 280 290 orfla . pep GVXSLSGD-VRHANDYGPMPIAGAAGDSGSPMFIYDKTNNKWLLNGVLQTGYPYSGRENG I : : I : :: :: I : II M : I : I II II II II I I :: I I I : I I I II I I II I : II orf1-1 GTVNLGSEKIKHS-PYGFLPTGGSFGDSGSPMFIYDAQKQKWLINGVLQTGNPYIGKSNG 250 260 270 280 290 300 310 320 330 340 350 orfla . pep FQLIRKDWFYDDIYRGDTHTVXFEPRSNGHFSFTSNNNGTGTVTETNEKVSNP-KLKVQT I I I: I I I I I I I: I : I I I I: I : I I I : I I :: I I::: I I I I I :: : I : I I : II:: I orf1-1 FQLVRKDWFYDEIFAGDTHSVFYEPRQNGKYSFNDDNNGTGKINAKHEHNSLPNRLKTRT 300 310 320 330 340 350 360 370 380 390 400 410 orfla . pep VRLFDESLNETDKEPVY-AAGGVNQYRPRLNNGENLSFIDYGNGKLILSNNINQGAGGLY IMM I |: II Mill I I I II I : I I I I II I II I : I II I I : I : I I I : : II I I I II I I I orf1-1 VQLFNVSLSETAREPVYHAAGGVNSYRPRLNNGENISFIDEGKGELILTSNINQGAGGLY 360 370 380 390 400 410 420 430 440 450 460 470 orfla . pep FEGDFTVSPENNETWQGAGVHISEDSTVTWKVNGVANDRLSKIGKGTLHVQAKGENQGSI I : I I II I I I I II I I I II II I I I I I I II I I I I I I I I I I I II I II I I I I I I I I II I I I I II I orf1-1 FQGDFTVSPENNETWQGAGVHISEDSTVTWKVNGVANDRLSKIGKGTLHVQAKGENQGSI 420 430 440 450 460 470 480 490 500 510 520 530 orfla . pep SVGDGTVILDQQADDKGKKQAFSEIGLXSGRGTVQLNADNQFNPDKLYFGFRGGRLDLNG I I I I II I I II II I II I I I II II II I I I I I II I I I II I I I II I II I I I I I I I II I I I I II orf1-1 SVGDGTVILDQQADDKGKKQAFSEIGLVSGRGTVQLNADNQFNPDKLYFGFRGGRLDLNG 480 490 500 510 520 530 540 550 560 570 580 590 orfla. pep HSLSFHRIQNTDEGAMIXXHNATTTSTVTITGNESITQPSGKNINRLNYSKEIAYNGWFG I II I I I II I I I I I I II I II I II II I II :: I : : \ : \ I I : : II I I I II I I I orf1-1 HSLSFHRIQNTDEGAMIVNHNQDKESTVTITGNKDIAT-TGNN-NSLDSKKEIAYNGWFG 540 550 560 570 580 590 600 610 620 630 640 650 orfla . pep EKDTTKTNGRLNLVYQPAAEDRTXLLSGGTNLNGNITQTNGKLFFSGRPTPHAYNHLGSG II I I II I I II I II II I I I I II I I I I II II I II I II II I II II I I II I I I II I I I I I : : orf1-1 EKDTTKTNGRLNLVYQPAAEDRTLLLSGGTNLNGNITQTNGKLFFSGRPTPHAYNHLNDH 600 610 620 630 640 650 660 670 680 690 700 710 orfla. pep WSKMEGIPQGEIVWDNDWIXRTFKAENFHIQGGQAVISRNVAKVEGDXHLSNHAQAVFGV II: I I II: II I II II I II I II I I II I : I : I I II I : I I I II I I : I I II II I II II I I I orf1-1 WSQKEGIPRGEIVWDNDWINRTFKAENFQIKGGQAWSRNVAKVKGDWHLSNHAQAVFGV 660 670 680 690 700 710 720 730 740 750 760 770 orfla . pep APHQSHTICTRSDWTGLTNCVEXXITDDKVIASLTKTDXSGXVXLXXXXXXXLXGXAXLX I I I I I I II I I I I I II I I I II II : I I II I II I II I I I I II I I I : I I: I orf1-1 APHQSHTICTRSDWTGLTNCVEKTITDDKVIASLTKTDISGNVDLADHAHLNLTGLATLN 720 730 740 750 760 770 780 790 800 810 820 830 orfla . pep GNLSANGDTRYTVSHNATQNGNLSLVGNAQATFNQATLNGNXSXSGNASFNLSNNAAQNG II I II I I I I I II I I II II I I I I I I I II I I I II I II I I I I I I : I I I I II I I I M: I: I I I orf1-1 GNLSANGDTRYTVSHNATQNGNLSLVGNAQATFNQATLNGNTSASGNASFNLSDHAVQNG 780 790 800 810 820 830 840 850 860 870 880 890 orfla . pep SLTLSDNAKANVSHSALNGNVSLADKAVFHFENSRFTGQLSGSKXTALHLKDSEWTLPSG I I I I I I I II I II I I II II I I II I I I I I I I II : I II II I : I I: I I I I I II I II I I I II I orf1-1 SLTLSGNAKANVSHSALNGNVSLADKAVFHFESSRFTGQISGGKDTALHLKDSEWTLPSG 840 850 860 870 880 890 900 910 920 930 940 orfla . pep TELGNLNLDNATITLNSAYRHDAAGAQTGXVSDTPRRRSRRS LLSVTPPTSVESRFN I I II II II II II II I I I I I II I II II II I :: IM I I I I I I I I I I I II I I I II II I I orf1-1 TELGNLNLDNATITLNSAYRHDAAGAQTGSATDAPRRRSRRSRRSLLSVTPPTSVESRFN 900 910 920 930 940 950 950 960 970 980 990 1000 orfla . pep TLTVNGKLNXQGTFRFMSELFGYRSDKLKLAESSEGTYTLAVNNTGNEPVSLDQLTWEG II I I I II II I I I I I I I I II I I I II I I I I I I II I I I II I I I I I I I II I I: I I: II II I I I orf1-1 TLTVNGKLNGQGTFRFMSELFGYRSDKLKLAESSEGTYTLAVNNTGNEPASLEQLTWEG 960 970 980 990 1000 1010 1010 1020 1030 1040 1050 1060 orfla . pep KDNKPLSENLNFTLQNEHVDAGAWRYQLIRKDGEFRLHNPVKEQELSDKLGKAEAKKQAE I I I II II I I I I II I I I II I II II I II II I II I I I I II II II I I II I I II I I II I I I I II I orf1-1 KDNKPLSENLNFTLQNEHVDAGAWRYQLIRKDGEFRLHNPVKEQELSDKLGKAEAKKQAE 1020 1030 1040 1050 1060 1070 1070 1080 1090 1100 1110 1120 orfla . pep KDNAQSLDALIAAGRDAAEKTESVAEPARXAGGENVGIMQAEEEKKRVQADKDSALAKQR I II I I I II II II I I I I I : I I I I I I I I I I I I II I II I I I I I II II I I I II II I : I I II I I orf1-1 KDNAQSLDALIAAGRDAVEKTESVAEPARQAGGENVGIMQAEEEKKRVQADKDTALAKQR 1080 1090 1100 1110 1120 1130 1130 1140 1150 1160 1170 1180 orfla . pep EAETRPXTTAFPRARXARRDLPQPQPQPQPQPQPQRDLXSRYANSGLSEFSATLNSVFAV I I II II I I I I I I I I I I I I I I I II II I I II I I I I I II II II I I I II I I I I II I I I orf1-1 EAETRPATTAFPRARRARRDLPQLQPQPQPQP—QRDLISRYANSGLSEFSATLNSVFAV 1140 1150 1160 1170 1180 1190 1190 1200 1210 1220 1230 1240 orfla . pep QDELDRVFAEDRRNAVWTSXIRXTKHYRSQDFRAYRQQTDLRQIGMQKNLGSGRVGILFS II I I I I I I I I I I I II II II II I I I I I II I I I I I I I II II I I I I I II I I I I I I II I I I I orf1-1 QDELDRVFAEDRRNAVWTSGIRDTKHYRSQDFRAYRQQTDLRQIGMQKNLGSGRVGILFS 1200 1210 1220 1230 1240 1250 1250 1260 1270 1280 1290 1300 orfla. pep HNRTENXFDDGIGNSARLAHGAVFGQYGIGRFDIGISTGAGFSSGXLSDGIGGKIRRRVL II I I I I : I I I I I I I I I I I II I II I II II I II II I I : II II I I I I I II II I I I I II II orf1-1 HNRTENTFDDGIGNSARLAHGAVFGQYGIDRFYIGISAGAGFSSGSLSDGIGGKIRRRVL 1260 1270 1280 1290 1300 1310 1310 1320 1330 1340 1350 1360 orfla . pep HYGIQARYRAGFGGFGIEPYIGATRYFVQKADYRYENVNIATPGLAFNRYRAGIKADYSF I I I II I I I I I I I II II II I : I I II I II II I I I I I II I I I I I II II I I I I I I I I I I I I II I orf1-1 HYGIQARYRAGFGGFGIEPHIGATRYFVQKADYRYENVNIATPGLAFNRYRAGIKADYSF 1320 1330 1340 1350 1360 1370 1370 1380 1390 1400 1410 1420 orfla . pep KPAQHXSITPYXSLSYTDAASGKVRTRVNTAVLAQDFGKTRSAEWGVNAEIKGFTLSXHA I I I I I I I I I I I I I I I I II I I I I II II I I I I I I I I II I I I I I I I I I I II II I I I I I II orf1-1 KPAQHISITPYLSLSYTDAASGKVRTRVNTAVLAQDFGKTRSAEWGVNAEIKGFTLSLHA 1380 1390 1400 1410 1420 1430 1430 1440 1450 orfla. pep AAAKGPQLEAQHSAGIKLGYRWX I I I I I I II I I I I I II I I I I I I II orfl-1 AAAKGPQLEAQHSAGIKLGYRWX 1440 1450 Homology with adhesion and penetration protein hap precursor of H.influenzae (accession number P45387) Amino acids 23-423 of ORFl show 59% aa identity with hap protein in 450aa overlap: orfl 23 FXAAYLAICLSFGILPQAWAGHTYFGINYQYYRDFAENKGKFAVGAKDIEVYNKKGELVG 82 F +L C+S GI QAWAGHTYFGI+YQYYRDFAENKGKF VGAK+IEVYNK+G+LVG hap 6 FRLNFLTACVSLGIASQAWAGHTYFGIDYQYYRDFAENKGKFTVGAKNIEVYNKEGQLVG 65 orfl 83 KSMTKAPMIDFSWSRNGVAALVGVQYIVSVAHNGGYNNVDFGAEGXNIXDQXRXTYKIV 142 SMTKAPMIDFSWSRNGVAALVG QYIVSVAHNGGYN+VDFGAEG N DQ R TY+IV hap 66 TSMTKAPMIDFSWSRNGVAALVGDQYIVSVAHNGGYNDVDFGAEGRN-PDQHRFTYQIV 124 orfl 143 KRNNYKAGTKGHPYGGDYHMPRLHKXVTDAEPVEMTSYMDGRKYIDQNNYPDRVRIGAGR 202 KRNNY+A + HPY GDYHMPRLHK VT+AEPV MT+ MDG+ Y D+ NYP+RVRIG+GR hap 125 KRNNYQAWERKHPYDGDYHMPRLHKFVTEAEPVGMTTNMDGKVYADRENYPERVRIGSGR 184 orfl 203 QYWRSDEDEPNNRESSYHIA 222 QYWR+D+DE N SSY+++ hap 185 QYWRTDKDEETNVHSSYYVSGAYRYLTAGNTHTQSGNGNGTVNLSGNWSPNHYGPLPTG 244 orfl 223 SGSPMFIYDAQKQKWLINGVLQTGNPYIGKSNGFQLVRKDWFYDEIFAGDTHSVF 277 SGSPMFIYDA+K++WLIN VLQTG+P+ G+ NGFQL+R++WFY+E+ A DT SVF hap 245 GSKGDSGSPMFIYDAKKKQWLINAVLQTGHPFFGRGNGFQLIREEWFYNEVLAVDTPSVF 304 orfl 278 —YEPRQNGKYSFNDDNNGTGKIN-AKHEHNSLPNRLKTRTVQLFNVSLSETAREPVYHA 334 Y P NG YSF +N+GTGK+ + + + + TV+LFN SL++TA+E V A hap 305 QRYIPPINGHYSFVSNNDGTGKLTLTRPSKDGSKAKSEVGTVKLFNPSLNQTAKEHV-KA 363 orfl 335 AGGVNSYRPRLNNGENISFIDEGKGELILTSNINQGAGGLYFQGDFTV-SPENNETWQGA 393 A G N Y+PR+ G+NI D+GKG L + +NINQGAGGLYF+G+F V +NN TWQGA hap 364 AAGYNIYQPRMEYGKNIYLGDQGKGTLTIENNINQGAGGLYFEGNFWKGKQNNITWQGA 423 orfl 394 GVHISEDSTVTWKVNGVANDRLSKIGKGTL 423 GV I +D+TV WKV+ NDRLSKIG GTL hap 424 GVSIGQDATVEWKVHNPENDRLSKIGIGTL 453 Amino acids 715-1011 of ORFl show 50% aa identity with hap protein in 258aa overlap: Orfl 41 DTRYTVSHNATQ-NGNXSLVXNAQATFNQ-ATLNGNTSASGNASFNLSDHAVQNGSLTLS 98 DT+ S TQ NG+ +L NA + A LNGN + ++ F LS++A Q G++ LS hap 733 DTKVINSIPITQINGSINLTNNATVNIHGLAKLNGNVTLIDHSQFTLSNNATQTGNIKLS 792 orfl 99 GNAKANVSHSALNGNVSLADKAVFHFESSRFTGQISGGKDTALHLKDSEWTLPSGXELGN 158 +A A V+++ LNGNV L D A F ++S F QI G KDT + L+++ WT+PS L N hap 793 NHANATVNNATLNGNVHLTDSAQFSLKNSHFWHQIQGDKDTTVTLENATWTMPSDTTLQN 852 orfl 159 LNLDNATITLNSAYRHDAAGAQTGSATDAPXXXXXXXXXXLLXVTPPTSVESRFNTLTVN 218 L L+N+T+TLNSAY + S+ +AP L T PTS E RFNTLTVN hap 853 LTLNNSTVTLNSAY SASSNNAPRHRRS LETETTPTSAEHRFNTLTVN 899 orfl 219 GKLNGQGTFRFMSELFGYRSDKLKLAESSEGTYTLAVNNTGNEPASLEQLTWEGKDNKP 278 GKL+GQGTF+F S LFGY+SDKLKL+ +EG YTL+V NTG EP +LEQLT++E DNKP hap 900 GKLSGQGTFQFTSSLFGYKSDKLKLSNDAEGDYTLΞVRNTGKEPVTLEQLTLIESLDNKP 959 orfl 279 LSENLNFTLQNEHVDAGA 296 LS+ L FTL+N+HVDAGA hap 960 LSDKLKFTLENDHVDAGA 977 Amino acids 1192-1450 of ORFl show 41% aa identity with hap protein in 259aa overlap: Orfl 1 LDRVFAEDRRNAVWTSGIRDTKHYRSQDFRAYRQQTDLRQIGMQKNLGSGRVGILFSHNR 60 LDR+F + ++AVWT+ +D + Y S FRAY+Q+T+LRQIG+QK L +GR+G +FSH+R hap 1135 LDRLFVDQAQSAVWTNIAQDKRRYDSDAFRAYQQKTNLRQIGVQKALANGRIGAVFSHSR 1194 orfl 61 TENTFDDGIGNSARLAHGAVFGQYGIDRFYXXXXXXXXXXXXXXXXXIGXKXRRRVLHYG 120 ++NTFD+ + N A L + F QY K R+ ++YG hap 1195 SDNTFDEQVKNHATLTMMSGFAQYQWGDLQFGVNVGTGISASKMAEEQSRKIHRKAINYG 1254 orfl 121 IQARYRAGFGGFGIEPHIGATRYFVQKADYRYENVNIATPGLAFNRYRAGIKADYSFKPA 180 + A Y+ G GI+P+ G RYF+++ +Y+ E V + TP LAFNRY AGI+ DY+F P hap 1255 VNASYQFRLGQLGIQPYFGVNRYFIERENYQSEEVRVKTPSLAFNRYNAGIRVDYTFTPT 1314 orfl 181 QHISITPYLSLSYTDAASGKVRTRVNTAVLAQDFGKTRSAEWGVNAEIKGFTLSLHAAAA 240 +IS+ PY ++Y D ++ V+T VN VL Q FG+ E G+ AEI F +S + + hap 1315 DNISVKPYFFVNYVDVSNANVQTTVNLTVLQQPFGRYWQKEVGLKAEILHFQISAFISKS 1374 orfl 241 KGPQLEAQHSAGIKLGYRW 259 +G QL Q + G+KLGYRW hap 1375 QGSQLGKQQNVGVKLGYRW 1393 Homology with a predicted ORF from N.gonorrhoeae The blocks of ORFl show 83.5%, 88.3%, and 97.7% identities in 467, 298, and 259 aa overlap, respectively with a predicted ORF (ORFlng) from N.gonorrhoeae: orfl . pep MKTTDKRTTETHRKAPKTGRIRFXAAYLAICLSFGILPQAWAGHTYFGINYQYYRDFAEN 60 I II I II I II II I I I I I II II I II II I II I I I I II I I I I I II II I I I II I II I I I I II orflng MKTTDKRTTETHRKAPKTGRIRFSPAYLAICLSFGILPQARAGHTYFGINYQYYRDFAEN 60 orfl . pep KGKFAVGAKDIEVYNKKGELVGKSMTKAPMIDFSWSRNGVAALVGVQYIVSVAHNGGYN 120 I I II I I I I I I I I I I II I I I II I I I I I I I II I I I II II I I II II I : I I I I I II I II II I I orflng KGKFAVGAKDIEVYNKKGELVGKSMTKAPMIDFSWSRNGVAALAGDQYIVSVAHNGGYN 120 orf1.pep NVDFGAEGXNIXDQXRXTYKIVKRNNYKAGTKGHPYGGDYHMPRLHKXVTDAEPVEMTSY 180 II I II II I I II I : I: II I I II I I I I I: I I II I II I I I II II I II I I I II I I I I I orflng NVDFGAEGSN-PDQHRFSYQIVKRNNYKAGTNGHPYGGDYHMPRLHKFVTDAEPVEMTSY 179 orfl . pep MDGRKYIDQNNYPDRVRIGAGRQYWRSDEDEPNNRESSYHIAS 223 III II I I : II I I I I I II I I II I I I I II II I I I II I II II I orflng MDGWKYADLNKYPDRVRIGAGRQYWRSDEDEPNNRESSYHIASAYSWLVGGNTFAQNGSG 239 orfl. pep GSPMFIYDAQKQKWLINGVLOTGNPYIGKSNG 255 I II I I II I II I I I I II I II I I I I I I II I I I I I orflng GGTVNLGSEKIKHSPYGFLPTGGSFGDSGSPMFIYDAQKQKWLINGVLOTGNPYIGKSNG 289 orf1.pep FQLVRKDWFYDEIFAGDTHSVFYEPRQNGKYSFNDDNNGTGKINAKHEHNSLPNRLKTRT 315 I M I I I I II I I I II I I I I I II I I I I : I I I I I II I : I I I : I II : I I I : I III I I I I II orflng FQLVRKDWFYDEIFAGDTHSVFYEPHQNGKYFFNDNNNGAGKIDAKHKHYSLPYRLKTRT 359 orfl . pep VQLFNVSLSETAREPVYHAAGGVNSYRPRLNNGENISFIDEGKGELILTSNINQGAGGLY 375 I I II I I I I I I II I II II II I I II I I I II II I II II I II I I: I II II I II I I I I II I II I I orflng VQLFNVSLSETAREPVYHAAGGVNSYRPRLNNGENISFIDKGKGELILTSNINQGAGGLY orfl. pep FQGDFTVSPENNETWQGAGVHISEDSTVTWKVNGVANDRLSKIGKGT 422 I : I : I II I I : I II I I I I I II I I I : I I II I I I I I I I I II II I I I I I I orflng FEGNFTVSPKNNETWQGAGVHISDGSTVTWKVNGVANDRLSKIGKGTLLVQAKGENQGSV 479 // orfl. pep DKVTASLTKTDISGNVDLADHAHLNLTGLA 744 III I I I : II I : I I I : I II I I I I I I I I I I orflng FGVAPHQSHTICTRSDWTGLTSCTEKTITDDKVIASLSKTDVRGNVSLADHAHLNLTGLA 774 orfl. pep TLNGNLSANGDTR-YTVSHNATQNGNXSLVXNAQATFNQATLNGNTSASGNASFNLSDHA 803 MMM :::: II : I I I I I II III I I I I I II II II II I I I II II I I II MM orflng TFNGNL-VQAETRTIRLRANATQNGNLSLVGNAQATFNQATLNGNTSASDNASFNLSNNA 833 orfl . pep VQNGSLTLSGNAKANVSHSALNGNVSLADKAVFHFESSRFTGQISGGKDTALHLKDSEWT 863 II I I I II I I I II I II II I II I I I I I I I I I II I I I I : I I I II M I I I II I I II II I M I I orflng VQNGSLTLSDNAKANVSHSALNGNVSLADKAVFHFENSRFTGKISGGKDTALHLKDSEWT 893 orfl .pep LPSGXELGNLNLDNATITLNSAYRHDAAGAQTGSATDAPRRRSRRSRRSLLXVTPPTSVE 923 I I II : I I I I I II I I I I I I I I I I I II I I I I I I I II I : I I II II I I I I II I II I II : I orflng LPSGTELGNLNLDNATITLNSAYRHDAAGAQTGSAADAPRRRSRRS LLSVTPPTSAE 950 orf1.pep SRFNTLTVNGKLNGQGTFRFMSELFGYRSDKLKLAESSEGTYTLAVNNTGNEPASLEQLT 983 I I I I I I I II I I I I II II I II I I I I I I I II I I II I I I I II II I I I I I I I I I II: I I II I I orflng SRFNTLTVNGKLNGQGTFRFMSELFGYRSGKLKLAESSEGTYTLAVNNTGNEPVSLEQLT 1010 orfl. pep WEGKDNKPLSENLNFTLQNEHVDAGAW 1011 II I I I II II I I I I II I II I II I I I I II orflng WEGKDNTPLSENLNFTLQNEHVDAGAWRYQLIRKDGEFRLHNPVKEQELSDKLGKAGET 1070 orfl . pep LDRVFAEDRRNAVWTSGIRDTKHYRSQDFR 1211 I I I I II I II I I I I II I I I I II I I I I I I I I I orflng PQRDLISRYANSGLSEFSATLNSVFAVQDELDRVFAEDRRNAVWTSGIRDTKHYRSQDFR 1239 orf1.pep AYRQQTDLRQIGMQKNLGSGRVGILFSHNRTENTFDDGIGNSARLAHGAVFGQYGIDRFY 1271 I M I I I I M I I I I II I II I II I I I I II I II I I II I I II I I I I I I I II I II I II I I II orflng AYRQQTDLRQIGMQKNLGSGRVGILFSHNRTGNTFDDGIGNSARLAHGAVFGQYGIGRFD 1299 orf1. pep IGISAGAGFSSGSLSDGIGXKXRRRVLHYGIQARYRAGFGGFGIEPHIGATRYFVQKADY 1331 I I I I I I I I I I I I II II I I I I I I I I II II II I I I II I II I II I I I I I I I I I II I I I I I orflng IGISAGAGFSSGSLSDGIRGKIRRRVLHYGIQARYRAGFGGFGIEPHIGATRYFVQKADY 1359 orfl. pep RYENVNIATPGLAFNRYRAGIKADYSFKPAQHISITPYLSLSYTDAASGKVRTRVNTAVL 1391 I I I I I I II I I I I II I I I I I I I I I I I I I II II II I I I II I II I I I I II I I II I I II I I I I I orflng RYENVNIATPGLAFNRYRAGIKADYSFKPAQHISITPYLSLSYTDAAΞGKVRTRVNTAVL 1419 orfl. pep AQDFGKTRSAEWGVNAEIKGFTLSLHAAAAKGPQLEAQHSAGIKLGYRW 1440 I I I I I I I II II II I II II I I I I I II I I I I I I I II I I I I I I I II I I I I I I orflng AQDFGKTRSAEWGVNAEIKGFTLSLHAAAAKGPQLEAQHSAGIKLGYRW 1468 The complete length ORFlng nucleotide sequence was identified <SEQ LD 653>: 1 ATGAAAACAA CCGACAAACG GACAACCGAA ACACACCGCA AAGCCCCTAA 51 AACCGGCCGC ATCCGCTTCT CGCCCGCTTA CTTAGCCATA TGCCTGTCGT 101 TCGGCATTCT GCCCCAAGCC CGGGCGGGAC ACACTTATTT CGGCATCAAC 151 TACCAATACT ATCGCGACTT TGCCGAAAAT AAAGGCAAGT TTGCAGTCGG 201 GGCGAAAGAT ATTGAGGTTT ACAACAAAAA AGGGGAGTTG GTCGGCAAAT 251 CGATGACGAA AGCCCCGATG ATTGATTTTT CTGTGGTATC GCGTAACGGC 301 GTGGCGGCAT TGGCGGGCGA TCAATATATT GTGAGCGTGG CACATAACGG 351 CGGCTATAAC AATGTTGATT TTGGTGCGGA GGGAAGCAAT CCCGATCAGC 401 ACCGCTTTTC TTACCAAATT GTGAAAAGAA ATAATTATAA AGCAGGGACT 451 AACGGCCATC CTTATGGCGG CGATTATCAT ATGCCGCGTT TGCACAAATT 501 TGTCACAGAT GCAGAACCTG TTGAGATGAC CAGTTATATG GATGGGTGGA 551 AATACGCTGA TTTAAATAAA TACCCTGATC GTGTTCGAAT CGGAGCAGGC 601 AGACAATATT GGCGGTCTGA TGAAGACGAA CCCAATAACC GCGAAAGTTC 651 ATATCATATT GCAAGCGCAT ATTCTTGGCT CGTCGGTGGC AATACCTTTG 701 CACAAAATGG ATCAGGTGGT GGCACAGTCA ACTTAGGTAG CGAAAAAATT 751 AAACATAGCC CATATGGTTT TTTACCAACA GGAGGCTCAT TTGGCGACAG 801 TGGCTCACCA ATGTTTATCT ATGATGCCCA AAAGCAAAAG TGGTTAATTA 851 ATGGGGTATT GCAAACAGGC AACCCCTATA TAGGAAAAAG CAATGGCTTC 901 CAGCTAGTTC GTAAAGATTG GTTCTATGAT GAAATCTTTG CTGGAGATAC 951 CCATTCAGTA TTCTACGAAC CACATCAAAA TGGGAAATAC TTTTTTAACG 1001 ACAATAATAA TGGCGCAGGA AAAATCGATG CCAAACATAA ACACTATTCT 1051 CTACCTTATA GATTAAAAAC ACGAACCGTT CAATTGTTTA ATGTTTCTTT 1101 ATCCGAGACA GCAAGAGAAC CTGTTTATCA TGCTGCAGGT GGGGTCAACA 1151 GTTATCGACC CAGACTGAAT AATGGAGAAA ATATTTCCTT TATTGACAAA 1201 GGAAAAGGTG AATTGATACT TACCAGCAAC ATCAACCAAG GCGCGGGCGG 1251 TTTGTATTTT GAGGGTAATT TTACGGTCTC GCCTAAAAAC AACGAAACGT 1301 GGCAAGGCGC GGGCGTTCAT ATCAGTGATG GCAGTACCGT TACTTGGAAA 1351 GTAAACGGCG TGGCAAACGA CCGCCTGTCC AAAATCGGCA AAGGCACGCT 1401 GCTGGTTCAA GCCAAAGGGG AAAACCAAGG CTCGGTCAGC GTGGGCGACG 1451 GTAAAGTCAT CTTAGATCAG CAGGCGGACG ATCAAGGCAA AAAACAAGCC 1501 TTTAGTGAAA TCGGCTTGGT CAGCGGCAGG GGGACGGTGC AACTGAATGC 1551 CGATAATCAG TTCAACCCCG ACAAACTCTA TTTCGGCTTT CGCGGCGGAC 1601 GTTTGGATTT GAACGGGCAT TCGCTTTCGT TCCACCGCAT TCAAAATACC 1651 GATGAAGGGG CGATGATTGT CAACCACAAT CAAGACAAAG AATCCACCGT 1701 TACCATTACA GGCAATAAAG ATATTACTAC AACCGGCAAT AACAACAACT 1751 TGGATAGCAA AAAAGAAATT GCCTACAACG GTTGGTTTGG CGAGAAAGAT 1801 GCAACCAAAA CGAACGGGCG GCTCAATCTG AATTACCAAC CGGAAGAAGC 1851 GGATCGCACT TTACTGCTTT CCGGCGGAAC AAATTTAAAC GGCAATATCA 1901 CGCAAACAAA CGGCAAACTG TTTTTCAGCG GCAGACCGAC ACCGCACGCC 1951 TACAATCATT TAGGAAGCGG GTGGTCAAAA ATGGAAGGTA TCCCACAAGG 2001 AGAAATCGTG TGGGACAACG ATTGGATCGA CCGCACATTT AAAGCGGAAA 2051 ACTTCCATAT TCAGGGCGGA CAAGCGGTGG TTTCCCGCAA TGTTGCCAAA 2101 GTGGAAGGCG ATTGGCATTT AAGCAATCAC GCCCAAGCAG TTTTCGGTGT 2151 CGCACCGCAT CAAAGCCACA CAATCTGTAC ACGTTCGGAC TGGACGGGTC 2201 TGACAAGTTG TACCGAAAAA ACCATTACCG ACGATAAAGT GATTGCTTCA 2251 TTGAGCAAGA CCGACATCAG AGGCAATGTC AGCCTTGCCG ATCACGCTCA 2301 TTTAAATCTC ACAGGACTTG CCACACTCAA CGGCAATCTT AGTGCAGGCG 2351 GAGACACGCA CTATACGGTT ACGCGCAACG CCACCCAAAA CGGCAACCTC 2401 AGCCTCGTGG GCAATGCCCA AGCAACATTT AATCAAGCCA CATTAAACGG 2451 CAACACATCG GCTTCGGACA ATGCTTCATT TAATCTAAGC AACAACGCCG 2501 TACAAAACGG CAGTCTGACG CTTTCCGACA ACGCTAAGGC AAACGTAAGC 2551 CATTCCGCAC TCAACGGCAA TGTCTCCCTA GCCGATAAGG CAGTATTCCA 2601 TTTTGAAAAC AGCCGCTTTA CCGGAAAAAT CAGCGGCGGC AAGGATACGG 2651 CATTACACTT AAAAGACAGC GAATGGACGC TGCCGTCGGG CACGGAATTA 2701 GGCAATTTAA ACCTTGACAA CGCCACCATT ACACTCAATT CCGCCTATCG 2751 ACACGATGCG GCAGGCGCGC AAACCGGCAG TGCGGCAGAT GCGCCGCGCC 2801 GCCGTTCGCG CCGTTCCCTA TTATCCGTTA CGCCGCCAAC TTCGGCAGAA 2851 TCCCGTTTCA ACACGCTGAC GGTAAACGGC AAATTGAACG GTCAGGGAAC 2901 ATTCCGCTTT ATGTCGGAAC TCTTCGGCTA CCGCAGCGGC AAATTGAAGC 2951 TGGCGGAAAG TTCCGAAGGC ACTTACACCT TGGCTGTCAA CAATACCGGC 3001 AACGAACCCG TAAGTCTCGA GCAATTGACG GTAGTGGAAG GAAAAGACAA 3051 CACACCGCTG TCCGAAAATC TTAATTTCAC CCTGCaaaAc gaacacgtcg 3101 atgccggcgc atggCGTTAT CAGCTTATCC gcaaagacgG CGAGTTCCgc 3151 CTGCATAATC CGGTCAAAGA ACAAGAGCTT TCCGACAAAC TCGGCAAGgc 3201 gggagaaACA GAggccgccT TGACGGCAAA ACAGGCacaA CTTGCCGCCA 3251 AAcaacaggc ggaaaAAGAC AACgcgcaaa gccttgAcgc gctgattgcg 3301 gCcgggcgca atgccaccga AAAGGCAgaa agtgttgccg aaccgGCCCG 33335511 GCAGGCAGGC GGGGAAAAtg ccgGCATTAT GCAGGCGGAG GAAGAGAAAA 3401 AACGGGTGCA GGCGGATAAA GACACCGCCT TGGCGAAACA GCGCGAAGCG 3451 GAAACCCGGC CGGCTACCAC CGCCTTCCCC CGCGCCCGCC GCGCCCGCCG 3501 GGATTTGCCG CAACCGCAGC CCCAACCGCA ACCCCAACCG CAGCGCGACC 3551 TGATCAGCCG TTATGCCAAT AGCGGTTTGA GTGAATTTTC CGCCACGCTC 33660011 AACAGCGTTT TCGCCGTACA GGACGAATTG GACCGCGTGT TTGCCGAAGA 3651 CCGCCGCAAC GCCGTTTGGA CAAGCGGCAT CCGGGACACC AAACACTACC 3701 GTTCGCAAGA TTTCCGCGCC TACCGCCAAC AAACCGACCT GCGCCAAATC 3751 GGTATGCAGA AAAACCTCGG CAGCGGGCGC GTCGGCATCC TGTTTTCGCA 3801 CAACCGGACC GGAAACACCT TCGACGACGG CATCGGCAAC TCGGCACGGC 33885511 TTGCCCACGG TGCCGTTTTC GGGCAATACG GCATCGGCAG GTTCGACATC 3901 GGCATCAGCG CGGGCGCGGG TTTTAGTAGC GGCAGCCTTT CAGACGGCAT 3951 CAGAGGCAAA ATCCGCCGCC GCGTGCTGCA TTACGGCATT CAGGCAAGAT 4001 ACCGCGCAGG TTTCGGCGGA TTCGGCATCG AACCGCACAT CGGCGCAACG 4051 CGCTATTTCG TCCAAAAAGC GGATTACCGA TACGAAAACG TCAATATCGC 44110011 CACCCCGGGC CTTGCATTCA ACCGCTACCG CGCGGGCATT AAGGCAGATT 4151 ATTCATTCAA ACCGGCGCAA CACATTTCCA TCACGCCTTA TTTGAGCCTG 4201 TCCTATACCG ATGCCGCTTC CGGCAAAGTC CGAACGCGCG TCAATACCGC 4251 CGTATTGGCG CAGGATTTCG GCAAAACCCG CAGTGCGGAA TGGGGCGTAA 4301 ACGCCGAAAT CAAAGGTTTC ACGCTGTCCC TCCACGCTGC CGCCGCCAAG 44335511 GGGCCGCAAT TGGAAGCGCA GCACAGCGCG GGCATCAAAT TAGGCTACCG 4401 CTGGTAA This is predicted to encode a protein having amino acid sequence <SEQ LD 654>: 1 MKTTDKRTTE THRKAPKTGR IRFSPAYLAI CLSFGILPQA RAGHTYFGIN 51 YQYYRDFAEN KGKFAVGAKD IEVYNKKGEL VGKSMTKAPM IDFSWSRNG 101 VAALAGDQYI VSVAHNGGYN NVDFGAEGSN PDQHRFSYQI VKRNNYKAGT 151 NGHPYGGDYH MPRLHKFVTD AEPVEMTSYM DGWKYADLNK YPDRVRIGAG 201 RQYWRSDEDE PNNRESSYHI ASAYSWLVGG NTFAQNGSGG GTVNLGSEKI 251 KHSPYGFLPT GGSFGDSGSP MFIYDAQKQK WLINGVLQTG NPYIGKSNGF 301 QLVRKDWFYD EIFAGDTHSV FYEPHQNGKY FFNDNNNGAG KIDAKHKHYS 351 LPYRLKTRTV QLFNVSLSET AREPVYHAAG GVNSYRPRLN NGENISFIDK 401 GKGELILTSN INQGAGGLYF EGNFTVSPKN NETWQGAGVH ISDGSTVTWK 451 VNGVANDRLS KIGKGTLLVQ AKGENQGSVS VGDGKVILDQ QADDQGKKQA 501 FSEIGLVSGR GTVQLNADNQ FNPDKLYFGF RGGRLDLNGH SLSFHRIQNT 551 DEGAMIVNHN QDKESTVTIT GNKDITTTGN NNNLDSKKEI AYNGWFGEKD 601 ATKTNGGLNL NYPPEEADRT LLLSGGTNLN GNITQTNGKL FFSGRPTPHA 651 YNHLGSGWSK MEGIPQGEIV WDNDWIDRTF KAENFHIQGG QAWSRNVAK 701 VEGDWHLSNH AQAVFGVAPH QSHTICTRSD WTGLTSCTEK TITDDKVIAS 751 LSKTDVRGNV SLADHAHLNL TGLATFNGNL VQAETRTIRL RANATQNGNL 801 SLVGNAQATF NQATLNGNTS ASDNASFNLS NNAVQNGSLT LSDNAKANVS 851 HSALNGNVSL ADKAVFHFEN SRFTGKISGG KDTALHLKDS EWTLPSGTEL 901 GNLNLDNATI TLNSAYRHDA AGAQTGSAAD APRRRSRRSL LSVTPPTSAE 951 SRFNTLTVNG KLNGQGTFRF MSELFGYRSG KLKLAESSEG TYTLAVNNTG 1001 NEPVSLEQLT EGKDNTPL SENLNFTLQN EHVDAGAWRY QLIRKDGEFR 1051 LHNPVKEQEL SDKLGKAGET EAALTAKQAQ LAAKQQAEKD NAQSLDALIA 1101 AGRNATEKAE SVAEPARQAG GENAGIMQAE EEKKRVQADK DTALAKQREA 1151 ETRPATTAFP RARRARRDLP QPQPQPQPQP QRDLISRYAN SGLSEFSATL 1201 NSVFAVQDEL DRVFAEDRRN AVWTSGIRDT KHYRSQDFRA YRQQTDLRQI 1251 GMQKNLGSGR VGILFSHNRT GNTFDDGIGN SARLAHGAVF GQYGIGRFDI 1301 GISAGAGFSS GSLSDGIRGK IRRRVLHYGI QARYRAGFGG FGIEPHIGAT 1351 RYFVQKADYR YENVNIATPG LAFNRYRAGI KADYSFKPAQ HIΞITPYLSL 1401 SYTDAASGKV RTRVNTAVLA QDFGKTRSAE WGVNAEIKGF TLSLHAAAAK 1451 GPQLEAQHSA GIKLGYRW* Underlined and double-underlined sequences represent the active site of a serine protease (trypsin family) and an ATP/GTP-binding site motif A (P-loop). ORFl-1 and ORFlng show 93.7% identity in 1471 aa overlap: 10 20 30 40 50 60 orf1-1. pep MKTTDKRTTETHRKAPKTGRIRFSPAYLAICLSFGILPQAWAGHTYFGINYQYYRDFAEN I II II I II I II I I I I I II II I I I II II I I I M M M I M I I I I I I II I II I II I I I II I orflng-1 MKTTDKRTTETHRKAPKTGRIRFSPAYLAICLSFGILPQARAGHTYFGINYQYYRDFAEN 10 20 30 40 50 60 70 80 90 100 110 120 orf1-1. pep KGKFAVGAKDIEVYNKKGELVGKSMTKAPMIDFSWSRNGVAALVGDQYIVSVAHNGGYN II I I II I I I I I II II II II II I I I II I I I II II II I I I I I I I I M I I I I I I II I I II I I I orflng-1 KGKFAVGAKDIEVYNKKGELVGKSMTKAPMIDFSWSRNGVAALAGDQYIVSVAHNGGYN 70 80 90 100 110 120 130 140 150 160 170 180 orf1-1. pep NVDFGAEGRNPDQHRFTYKIVKRNNYKAGTKGHPYGGDYHMPRLHKFVTDAEPVEMTSYM II I I II I I I I I I II I : I : I II II II II IM I I I I I I II I I I II I II I I I I I I I I I I I II orflng-1 NVDFGAEGSNPDQHRFSYQIVKRNNYKAGTNGHPYGGDYHMPRLHKFVTDAEPVEMTSYM 130 140 150 160 170 180 190 200 210 220 230 240 orf1-1.pep DGRKYIDQNNYPDRVRIGAGRQYWRSDEDEPNNRESSYHIASAYSWLVGGNTFAQNGSGG II II I I : II I I II II I I I I I II I I I I II II II II I I I II I I I II I I I I I I I I I I I II orflng-1 DGWKYADLNKYPDRVRIGAGRQYWRSDEDEPNNRESSYHIASAYSWLVGGNTFAQNGSGG 190 200 210 220 230 240 250 260 270 280 290 300 orf1-1. pep GTVNLGSEKIKHSPYGFLPTGGSFGDSGSPMFIYDAQKQKWLINGVLQTGNPYIGKSNGF II I II II I II II I I I I II I II II I II II II II II II I I I II I I I II I I I I II I I I II I I I orflng-1 GTVNLGSEKIKHSPYGFLPTGGSFGDSGSPMFIYDAQKQKWLINGVLQTGNPYIGKSNGF 250 260 270 280 290 300 310 320 330 340 350 360 orfl-l.pep QLVRKDWFYDEIFAGDTHSVFYEPRQNGKYSFNDDNNGTGKINAKHEHNSLPNRLKTRTV I I II I I II I II I I II I I I I I I I II : I I I I I II I : II I : I I I : I I I : I III I I I I I II orflng-1 QLVRKDWFYDEIFAGDTHSVFYEPHQNGKYFFNDNNNGAGKIDAKHKHYSLPYRLKTRTV 310 320 330 340 350 360 370 380 390 400 410 420 orfl-l.pep QLFNVSLSETAREPVYHAAGGVNSYRPRLNNGENISFIDEGKGELILTSNINQGAGGLYF I I I I II I II I II I I II II II I II I II II II II II II I I I : I II I II I II I I II I I II II I orflng-1 QLFNVSLSETAREPVYHAAGGVNSYRPRLNNGENISFIDKGKGELILTSNINQGAGGLYF 370 380 390 400 410 420 430 440 450 460 470 480 orfl-l.pep QGDFTVSPENNETWQGAGVHISEDSTVTWKVNGVANDRLSKIGKGTLHVQAKGENQGSIS : I : I II I I : I I I II II II I I I I : II II I I I I I I II II II I I I II I I I II I II I II I : I orflng-1 EGNFTVSPKNNETWQGAGVHISDGSTVTWKVNGVANDRLSKIGKGTLLVQAKGENQGSVS 430 440 450 460 470 480 490 500 510 520 530 540 orf1-1.pep VGDGTVILDQQADDKGKKQAFSEIGLVSGRGTVQLNADNQFNPDKLYFGFRGGRLDLNGH II I I I I I II I II I : I I I I I I I I II I II II II II II II I I I I I I II I I I II I II I II I I I orflng-1 VGDGKVILDQQADDQGKKQAFSEIGLVSGRGTVQLNADNQFNPDKLYFGFRGGRLDLNGH 490 500 510 520 530 540 550 560 570 580 590 600 orfl-l.pep SLSFHRIQNTDEGAMIVNHNQDKESTVTITGNKDIATTGNNNSLDSKKEIAYNGWFGEKD I I I I I II I I I I I I I I II II II I II I I I I I I I I II I : II I I I I : I II I I I II II I I I II I I orflng-1 SLSFHRIQNTDEGAMIVNHNQDKESTVTITGNKDITTTGNNNNLDSKKEIAYNGWFGEKD 550 560 570 580 590 600 610 620 630 640 650 660 orfl-l.pep TTKTNGRLNLVYQPAAEDRTLLLSGGTNLNGNITQTNGKLFFSGRPTPHAYNHLNDHWSQ : I I I I II I II III I II I I I I II I II II I I II II II I I I II I I I II I I II I : : I orflng-1 ATKTNGRLNLNYQPEEADRTLLLSGGTNLNGNITQTNGKLFFSGRPTPHAYNHLGSGWSK 610 620 630 640 650 660 670 680 690 700 710 720 orf1-1. pep KEGIPRGEIVWDNDWINRTFKAENFQIKGGQAWSRNVAKVKGDWHLSNHAQAVFGVAPH II I I : I II I I I I I I I : II I I I II I: I : II II I I I I I I II M I I I I II I I II I I II I II I orflng-1 MEGIPQGEIVWDNDWIDRTFKAENFHIQGGQAWSRNVAKVEGDWHLSNHAQAVFGVAPH 670 680 690 700 710 720 730 740 750 760 770 780 orfl-l.pep QSHTICTRSDWTGLTNCVEKTITDDKVIASLTKTDISGNVDLADHAHLNLTGLATLNGNL I II I II I I II II I I I : I: II I II II I II II I : II II II I : I I I I I I I I II II II I I I II orflng-1 QSHTICTRSDWTGLTSCTEKTITDDKVIASLSKTDIRGNVSLADHAHLNLTGLATLNGNL 730 740 750 760 770 780 790 800 810 820 830 840 orf1-1. pep SANGDTRYTVSHNATQNGNLSLVGNAQATFNQATLNGNTSASGNASFNLSDHAVQNGSLT II: II I: I I I :: II I I II I I I I I II I I I I II I II II I II I II II I I I I I :: II I I I I II orflng-1 SAGGDTHYTVTRNATQNGNLSLVGNAQATFNQATLNGNTSASDNASFNLSNNAVQNGSLT 790 800 810 820 830 840 850 860 870 880 890 900 orfl-l.pep LSGNAKANVSHSALNGNVSLADKAVFHFESSRFTGQISGGKDTALHLKDSEWTLPSGTEL II I II I I II II I II I II I II I II I I I I I : I II I M I I I I I I I I I I I I I II II II I I I II orflng-1 LSDNAKANVSHSALNGNVSLADKAVFHFENSRFTGKISGGKDTALHLKDSEWTLPSGTEL 850 860 870 880 890 900 910 920 930 940 950 960 orf1-1.pep GNLNLDNATITLNSAYRHDAAGAQTGSATDAPRRRSRRSRRSLLSVTPPTSVESRFNTLT I I I I II I I I I II I I I I I I II I I I I I I I I : I I II I I I I II I II I I I I I I : I II II I I I orflng-1 GNLNLDNATITLNSAYRHDAAGAQTGSAADAPRRRSR RSLLSVTPPTSAESRFNTLT 910 920 930 940 950 970 980 990 1000 1010 1020 orfl-l.pep VNGKLNGQGTFRFMSELFGYRSDKLKLAESSEGTYTLAVNNTGNEPASLEQLTWEGKDN I I I II I II I II I II I I I II I I I I II II II I II I I II II I II I I I I : I I I I I II II I I II orflng-1 VNGKLNGQGTFRFMSELFGYRSGKLKLAESSEGTYTLAVNNTGNEPVSLEQLTWEGKDN 960 970 980 990 1000 1010 1030 1040 1050 1060 1070 orfl-l.pep KPLSENLNFTLQNEHVDAGAWRYQLIRKDGEFRLHNPVKEQELSDKLGKA II I II I I II I II I I I I II I I I I I II II I II II I II I II II I I I I I I I I I orflng-1 TPLSENLNFTLQNEHVDAGAWRYQLIRKDGEFRLHNPVKEQELSDKLGKAGETEAALTAK 1020 1030 1040 1050 1060 1070 1080 1090 1100 1110 1120 orf1-1. pep EAKKQAEKDNAQSLDALIAAGRDAVEKTESVAEPARQAGGENVGIMQAEEEKKRVQ I I: I I II I II I II I I II I II I : I: I I : I I I I I II I II I II I : I II II I I II II I I orflng-1 QAQLAAKQQAEKDNAQSLDALIAAGRNATEKAESVAEPARQAGGENAGIMQAEEEKKRVQ 1080 1090 1100 1110 1120 1130 1130 1140 1150 1160 1170 1180 orfl-l.pep ADKDTALAKQREAETRPATTAFPRARRARRDLPQLQPQPQPQPQRDLISRYANSGLSEFS I II I II I I II I I II I II II I I II I I II II II I II I II I I I I I I II I I I II II II II I II orflng-1 ADKDTALAKQREAETRPATTAFPRARRARRDLPQPQPQPQPQPQRDLISRYANSGLSEFS 1140 1150 1160 1170 1180 1190 1190 1200 1210 1220 1230 1240 orf1-1.pep ATLNΞVFAVQDELDRVFAEDRRNAVWTSGIRDTKHYRSQDFRAYRQQTDLRQIGMQKNLG I I I I I II I II I I II I I I I II I I II I II I I I II I II II I II II I II I I I II II I II II I I I orflng-1 ATLNSVFAVQDELDRVFAEDRRNAVWTSGIRDTKHYRSQDFRAYRQQTDLRQIGMQKNLG 1200 1210 1220 1230 1240 1250 1250 1260 1270 1280 1290 1300 orfl-l.pep SGRVGILFSHNRTENTFDDGIGNSARLAHGAVFGQYGIDRFYIGISAGAGFSSGSLSDGI I I I I I I II I I I I I I I I I I I II I II I I I I II II I I I II II I I II I I I I I II I I I I I II orflng-1 SGRVGILFSHNRTGNTFDDGIGNSARLAHGAVFGQYGIGRFDIGISAGAGFSSGSLSDGI 1260 1270 1280 1290 1300 1310 1310 1320 1330 1340 1350 1360 orfl-l.pep GGKIRRRVLHYGIQARYRAGFGGFGIEPHIGATRYFVQKADYRYENVNIATPGLAFNRYR II I I II I II I I II I II I II I I I II I II I I I I II I I II I I I I I I II I I I I II I II II I I I orflng-1 RGKIRRRVLHYGIQARYRAGFGGFGIEPHIGATRYFVQKADYRYENVNIATPGLAFNRYR 1320 1330 1340 1350 1360 1370 1370 1380 1390 1400 1410 1420 orf1-1. pep AGIKADYSFKPAQHISITPYLSLSYTDAASGKVRTRVNTAVLAQDFGKTRSAEWGVNAEI II I II I I I I I I I II I II I II I I II I I I I I I I I II II I I I I I I I I I I I I II II I I II I I I I orflng-1 AGIKADYSFKPAQHISITPYLSLSYTDAASGKVRTRVNTAVLAQDFGKTRSAEWGVNAEI 1380 1390 1400 1410 1420 1430 1430 1440 1450 orf1-1. pep KGFTLSLHAAAAKGPQLEAQHSAGIKLGYRWX II II II II I I II I II II II I II II II I I II I I orflng-1 KGFTLSLHAAAAKGPQLEAQHSAGIKLGYRWX 1440 1450 1460 ition, ORFlng shows 55.7% identity with hap protein (P45387) over a 1455aa overlap: SCORES Initl: 1104 Initn: 4632 Opt: 2680 Smith-Waterman score: 5165; 55.7% identity in 1455 aa overlap 10 20 30 40 50 60 orflng-1. pep MKTTDKRTTETHRKAPKTGRIRFSPAYLAICLSFGILPQARAGHTYFGINYQYYRDFAEN I : I : IMMM I I II II I I I : II II II I I II p45387 MKKTVFRLNFLTACISLGIVSQAWAGHTYFGIDYQYYRDFAEN 10 20 30 40 70 80 90 100 110 120 orflng-1. pep KGKFAVGAKDIEVYNKKGELVGKSMTKAPMIDFSWSRNGVAALAGDQYIVSVAHNGGYN II I I: I I I :: I: I I I I: I: I II M II II I II I I I II II II II M : I I I I I I I I I II: p45387 KGKFTVGAQNIKVYNKQGQLVGTSMTKAPMIDFSWSRNGVAALVENQYIVSVAHNVGYT 50 60 70 80 90 100 130 140 150 160 170 180 orflng-l.pep NVDFGAEGSNPDQHRFSYQIVKRNNYKAGTNGHPYGGDYHMPRLHKFVTDAEPVEMTSYM : I I II I II : I II II II : I : II II I II I I II I I II II I II I II M M M I I p45387 DVDFGAEGNNPDQHRFTYKIVKRNNYKKD-NLHPYEDDYHNPRLHKFVTEAAPIDMTSNM 110 120 130 140 150 160 190 200 210 220 230 240 orflng-1.pep DGWKYADLNKYPDRVRIGAGRQYWRSDEDEPNNRESSYHIASAYSWLVGGNTFAQNGSGG : I I : I : I II : I I I I I : I II : I I : I M : : : : I : I I : I : : I I I I MM p45387 NGSTYSDRTKYPERVRIGSGRQFWRNDQDKGD QVAGAYHYLTAGNTHNQRGAGN 170 180 190 200 210 250 260 270 280 290 300 orflng-1. pep GTVNLGSEKIKHSPYGFLPTGGSFGDSGSPMFIYDAQKQKWLINGVLQTGNPYIGKSNGF I I I :: I : II II Ml II I I I II I I II I : I I I I II I I : I : M II III p45387 GYSYLGGDVRKAGEYGPLPIAGSKGDSGSPMFIYDAEKQKWLINGILREGNPFEGKENGF 220 230 240 250 260 270 310 320 330 340 350 360 orflng-1. pep QLVRKDWFYDEIFAGDTHSVFYEPHQNGKYFFNDNNNGAGKIDAKHKHYSLPYRLKTRTV I I I II :: I I I I I I M I I I : : I : II I : I I : : I :: I : p45387 QLVRKSYF-DEIFERDLHTSLYTRAGNGVYTISGNDNGQGSITQKS GIPSEIK I 280 290 300 310 320 370 380 390 400 410 419 orflng-l.pep QLFNVSLSETAREPVYHAA-GGVNSYRPRLNNGENISFIDKGKGELILTSNINQGAGGLY I MM :: I :: I I I I II I I II:: MM : I I I :: I : I I I I I I II I p45387 TLANMSLPLKEKDKVHNPRYDGPNIYSPRLNNGETLYFMDQKQGSLIFASDINQGAGGLY 330 340 350 360 370 380 420 430 440 450 460 470 479 orflng-l.pep FEGNFTVSPKNNETWQGAGVHISDGSTVTWKVNGVANDRLSKIGKGTLLVQAKGENQGSV I I II I I I II :: I : I II II I : I : I :: I I I I I II I II : I I I I I II I I II I I II II I : I I : p45387 FEGNFTVSPNSNQTWQGAGIHVSENSTVTWKVNGVEHDRLSKIGKGTLHVQAKGENKGSI 390 400 410 420 430 440 480 490 500 510 520 530 539 orflng-1.pep SVGDGKVILDQQADDQGKKQAFSEIGLVSGRGTVQLNADNQFNPDKLYFGFRGGRLDLNG I I I I II I II : I I I I I I I : II I I II II I II I II I I II I IMM I I : I M I I II II II II p45387 ΞVGDGKVILEQQADDQGNKQAFSEIGLVSGRGTVQLNDDKQFDTDKFYFGFRGGRLDLNG 450 460 470 480 490 500 540 550 560 570 580 590 orflng-1. pep HSLSFHRIQNTDEGAMIVNHNQDKESTVTITGNKDITT-TGNN-NNLDSKKEIAYNGWFG I I I: I : I I I I I I I I II I I I II : :: II I I II :: I : MM I: I I : II I I I I I II I p45387 HSLTFKRIQNTDEGAMIVNHNTTQAANVTITGNESIVLPNGNNINKLDYRKEIAYNGWFG 510 520 530 540 550 560 600 610 620 630 640 650 orflng-1. pep EKDATKTNGRLNLNYQPEEADRTLLLSGGTNLNGNITQTNGKLFFSGRPTPHAYNHLGSG I I : I II I I I I Ml I I II I I II II II : I : I I II : I I I I I I I I I II I I I I I I : : p45387 ETDKNKHNGRLNLIYKPTTEDRTLLLSGGTNLKGDITQTKGKLFFSGRPTPHAYNHLNKR 570 580 590 600 610 620 660 670 680 690 700 710 orflng-l.pep WSKMEGIPQGEIVWDNDWIDRTFKAENFHIQGGQAWSRNVAKVEGDWHLSNHAQAVFGV I I : II I II I II II II : II I : I II I II II : I : I I : I I I II I I ::: I I : I : IM |: |: II I p45387 WSEMEGIPQGEIVWDHDWINRTFKAENFQIKGGSAWSRNVSSIEGNWTVSNNANATFGV 630 640 650 660 670 680 720 730 740 750 760 770 orflng-1. pep APHQSHTICTRSDWTGLTSCTEKTITDDKVIASLSKTDIRGNVSLADHAHLNLTGLATLN : I: I :: I I II II II I II I : I : : II I II I : II : I I : : : I : I : I M I I I I I p45387 VPNQQNTICTRSDWTGLTTCQKVDLTDTKVINSIPKTQINGSINLTDNATANVKGLAKLN 690 700 710 720 730 740 780 790 800 810 820 830 orflng-1.pep GNLSAGGDTHYTVTRNATQNGNLSLVGNAQATFNQATLNGNTSASDNASFNLSNNAVQNG II:: ::::: |: I I I I I: I I p45387 GNVTL TNHSQFTLSNNATQIG 750 760 770 840 850 860 870 880 890 orflng-1. pep SLTLSDNAKANVSHSALNGNVSLADKAVFHFENSRFTGKISGGKDTALHLKDSEWTLPSG :: I I IM MM:: II II I I : |: I I :: I M I : : I : I I I :: I ::: I M p45387 NIRLSDNSTATVDNANLNGNVHLTDSAQFSLKNSHFSHQIQGDKGTTVTLENATWTMPSD 780 790 800 810 820 830 900 910 920 930 940 950 orflng-l.pep TELGNLNLDNATITLNSAYRHDAAGAQTGSAADAPRRRSRRSLLSVTPPTSAESRFNTLT I I I I : I : I : I I II I I I I : : I : :: I II I I I : I II II I I I I II I p45387 TTLQNLTLNNSTITLNSAY SASSNNTPRRRS LETETTPTSAEHRFNTLT 840 850 860 870 960 970 980 990 1000 1010 orflng-1. pep VNGKLNGQGTFRFMSELFGYRSGKLKLAESSEGTYTLAVNNTGNEPVSLEQLTWEGKDN I II I I : II I I I : I I MMM I II I :: :: I I I I : I MMM : I II I I : I I : II I p45387 VNGKLSGQGTFQFTSSLFGYKSDKLKLSNDAEGDYILSVRNTGKEPETLEQLTLVESKDN 880 890 900 910 920 930 1020 1030 1040 1050 1060 1070 orflng-l.pep TPLSENLNFTLQNEHVDAGAWRYQLIRKDGEFRLHNPVKEQELSDKLGKAGETEAALTAK I I I : : I : II I : I : II II I I I I : I : : : II II I I II I : I I I I I : I : I : : I : I I I p45387 QPLSDKLKFTLENDHVDAGALRYKLVKNDGEFRLHNPIKEQELHNDLVRAEQAERTLEAK 940 950 960 970 980 990 1080 1090 1100 1110 1120 1130 orflng-1.pep QAQLAAKQQAEKDNAQSLDALIAAGRNAT-EKAESVAEPARQAGGENAGIMQAEEEKKRV I :: : I I M : :: : I I II : : :: : I MM : I :::: : I : I p45387 QVEPTAKTQTGEPKVRSRRAARAAFPDTLPDQSLLNALEAKQAE-LTAETQKSKAKTKKV 1000 1010 1020 1030 1040 1050 1140 1150 1160 1170 1180 1190 orflng-l.pep QADK DTALAKQREAETRPATTAFPRARRARRD-LPQPQPQPQPQPQRDLISRYANSG :: : : I I : I :: : : : : : I I I : : I : I : I I I I I I : I I : p45387 RSKRAVFSDPLLDQSLFALEAALEVIDAPQQSEKDRLAQEEAEKQ-RKQKDLISRYSNSA 1060 1070 1080 1090 1100 1110 1200 1210 1220 1230 1240 1250 orflng-l.pep LSEFSATLNSVFAVQDELDRVFAEDRRNAVWTSGIRDTKHYRSQDFRAYRQQ-TDLRQIG I I I : I I I: I I::: I I I I II I: I::: :: I II M : I :: I I : I I I M I I M I II I I p45387 LSELSATVNSMLSVQDELDRLFVDQAQSAVWTNIAQDKRRYDSDAFRAYQQQKTNLRQIG 1120 1130 1140 1150 1160 1170 1260 1270 1280 1290 1300 1310 orflng-1. pep MQKNLGSGRVGILFSHNRTGNTFDDGIGNSARLAHGAVFGQYGIGRFDIGISAGAGFSSG : I I I :: II : I : I I I : I : IMM : I I I : : MM I : : : I : : : I : I : I : : p45387 VQKALANGRIGAVFSHSRSDNTFDEQVKNHATLTMMSGFAQYQWGDLQFGVNVGTGISAS 1180 1190 1200 1210 1220 1230 1320 1330 1340 1350 1360 1370 or lng-1. pep SLSDGIRGKIRRRVLHYGIQARYRAGFGGFGIEPHIGATRYFVQKADYRYENVNIATPGL : : : : II : I : : : : II : : I I : : I : I I : I : : I : : II I : : : : I : I : I : M p45387 KMAEEQSRKIHRKAINYGVNASYQFRLGQLGIQPYFGVNRYFIERENYQSEEVRVKTPSL 1240 1250 1260 1270 1280 1290 1380 1390 1400 1410 1420 1430 orflng-1. pep AFNRYRAGIKADYSFKPAQHISITPYLSLSYTDAASGKVRTRVNTAVLAQDFGKTRSAEW I I II I I I I:: I I : I I ::: I I : II: : : I : I : : : : : I : I II : | | I IM : I p45387 AFNRYNAGIRVDYTFTPTDNISVKPYFFVNYVDVSNANVQTTVNLTVLQQPFGRYWQKEV 1300 1310 1320 1330 1340 1350 1440 1450 1460 1469 orflng-1.pep GVNAEIKGFTLSLHAAAAKGPQLEAQHSAGIKLGYRWX MMM I : I : : : I I I I : :: I : I I I I I I p45387 GLKAEILHFQISAFISKSQGSQLGKQQNVGVKLGYRW 1360 1370 1380 1390 Based on this analysis, it is predicted that these proteins from N. meningitidis .and N.gonorrhoeae, and their epitopes, could be useful .antigens for vaccines or diagnostics, or for raising antibodies. Example 78 The following partial DNA sequence was identified in N. meningitidis <SEQ ID 655>: 1 ..AAGGTGTGGC AATTTGTCGA AGA.CCGCTG CGTGCCGTCG TGCCTGCCGA 51 CAGTTTTGAA CCGACCGCGC AAAAATTGAA CCTGTTTAAG GCGGGTGCGG 101 CAACCATTTT GTTTTATGAA GATCAAAATG TCGTCAAAGG TTTGCAGGAG 151 CAGTTCCCTG CTTATGCCGC TAACTTCCCC GTTTGGGCGg ATCAGGCAAA 201 CGCGATGGTG CAGTATGCCG TTTGGACGAC ACTTGCCGCG GTCGGCGTAG 251 GTGCAAACCT GCAACATTAC AATCCCTTGC CCGATGCGGC GATTGCCAAA 301 GCGTGGAATA TCCCCGAAAA CTGGTTGTTG CGCGCACAAA TGGTTATCGG 351 CGGTATTGAA GGGGCGGCAG GTGAAAAGAC CTTTGAACCC GTTGCAGAAC 401 GTTTGAAAGT GTTCGGCGCA TAA This coπesponds to the amino acid sequence <SEQ ID 656; ORF6>: 1 ..KVWQFVEXPL RAWPADSFE PTAQKLNLFK AGAATILFYE DQNWKGLQE 51 QFPAYAANFP VWADQANAMV QYAVWTTLAA VGVGANLQHY NPLPDAAIAK 101 AWNIPENWLL RAQMVIGGIE GAAGEKTFEP VAERLKVFGA * Further sequence analysis revealed a further partial DNA sequence <SEQ LD 657>: 1 .. CTGCGTGCCG TCGTGCCTGC CGACAGTTTT GAACCGACCG CGCAAAAATT 51 GAACCTGTTT AAGGCGGGTG CGGCAACCAT TTTGTTTTAT GAAGATCAAA 101 ATGTCGTCAA AGGTTTGCAG GAGCAGTTCC CTGCTTATGC CGCTAACTTC 151 CCCGTTTGGG CGGATCAGGC AAACGCGATG GTGCAGTATG CCGTTTGGAC 201 GACACTTGCC GCGGTCGGCG TAGGTGCAAA CCTGCAACAT TACAATCCCT 251 TGCCCGATGC GGCGATTGCC AAAGCGTGGA ATATCCCCGA AAACTGGTTG 301 TTGCGCGCAC AAATGGTTAT CGGCGGTATT GAAGGGGCGG CAGGTGAAAA 351 GACCTTTGAA CCCGTTGCAG AACGTTTGAA AGTGTTCGGC GCATAA This coπesponds to the amino acid sequence <SEQ LD 658; ORF6-l>: 1 ..LRAWPADSF EPTAQKLNLF KAGAATILFY EDQNWKGLQ EQFPAYAANF 51 PVWADQANAM VQYAVWTTLA AVGVGANLQH YNPLPDAAIA KAWNIPENWL 101 LRAQMVIGGI EGAAGEKTFE PVAERLKVFG A* Computer analysis of this amino acid sequence gave the following results: Homology with a predicted ORF from N.meninsitidis (sfrain Al ORF6 shows 98.6% identity over a 140aa overlap with an ORF (ORF6a) from strain A of N meningitidis: 10 20 30 orf6. pep KVWQFVEXPLRAWPADSFEPTAQKLNLFK I II I II I I I I II I I II I I II II I II I I I orf6a QIVEHAVLHTPSSFNSQSARVWLFGEEHDKVWQFVEDALRAWPADSFEPTAQKLNLFK 40 50 60 70 80 90 40 50 60 70 80 90 orf6. pep AGAATILFYEDQNVVKGLQEQFPAYAANFPVWADQANAMVQYAVWTTLAAVGVGANLQHY I I II I I II I II II I II I I II I II II I I II I I II I II II I I I I II I I II I I II I II II II I orf6a AGAATILFYEDQNWKGLQEQFPAYAANFPVWADQANAMVQYAVWTTLAAVGVGANLQHY 100 110 120 130 140 150 100 110 120 130 140 orf6. pep NPLPDAAIAKAWNIPENWLLRAQMVIGGIEGAAGEKTFEPVAERLKVFGAX I II I II II II I I I I II II II I I I I I I I I II I I I I I I I I I I I I I II II I I I I orf6a NPLPDAAIAKAWNIPENWLLRAQMVIGGIEGAAGEKTFEPVAERLKVFGAX 160 170 180 190 200 The complete length ORF6a nucleotide sequence <SEQ ID 659> is: 1 ATGACCCGTC AATCTCTGCA ACAGGCTGCC GAAAGCCGCC GTTCCATTTA 51 TTCGTTAAAT AAAAATCTGC CCGTCGGCAA AGATGAAATC GTCCAAATCG 101 TCGAACACGC CGTTTTGCAC ACACCTTCTT CGTTCAATTC CCAATCTGCC 151 CGTGTGGTCG TGCTGTTTGG CGAAGAGCAT GATAAGGTGT GGCAATTTGT 201 CGAAGACGCG CTGCGTGCCG TCGTGCCTGC CGACAGTTTT GAACCGACCG 251 CGCAAAAATT GAACCTGTTT AAGGCGGGTG CGGCAACTAT TTTGTTTTAT 301 GAAGATCAAA ATGTCGTCAA AGGTTTGCAG GAGCAGTTCC CTGCTTATGC 351 CGCCAACTTT CCCGTTTGGG CGGACCAGGC GAACGCGATG GTGCAGTATG 401 CCGTTTGGAC GACACTTGCC GCGGTCGGCG TAGGTGCAAA CCTGCAACAT 451 TACAATCCCT TGCCCGATGC GGCGATTGCC AAAGCGTGGA ATATCCCCGA 501 AAACTGGTTG TTGCGCGCAC AAATGGTTAT CGGCGGTATT GAAGGGGCGG 551 CAGGTGAAAA GACCTTTGAA CCAGTTGCAG AACGTTTGAA AGTGTTCGGC 601 GCATAA This is predicted to encode a protein having amino acid sequence <SEQ ID 660>: 1 MTRQSLQQAA ESRRSIYSLN KNLPVGKDEI VQIVEHAVLH TPSSFNSQSA 51 RWVLFGEEH DKVWQFVEDA LRAWPADSF EPTAQKLNLF KAGAATILFY 101 EDQNWKGLQ EQFPAYAANF PVWADQANAM VQYAVWTTLA AVGVGANLQH 151 YNPLPDAAIA KAWNIPENWL LRAQMVIGGI EGAAGEKTFE PVAERLKVFG 201 A* ORF6a and ORF6-1 show 100.0% identity in 131 aa overlap: 50 60 70 80 90 100 orf6a. pep TPSSFNSQSARWVLFGEEHDKVWQFVEDALRAWPADSFEPTAQKLNLFKAGAATILFY II I II I I I I II I I I I I I I I I I I I I I I I I I I orf6-1 LRAWPADΞFEPTAQKLNLFKAGAATILFY 10 20 30 110 120 130 140 150 160 orf6a . pep EDQNWKGLQEQFPAYAANFPVWADQANAMVQYAVWTTLAAVGVGANLQHYNPLPDAAIA I I I I I I I I I II II I I II II I I I II I I II I II I I I II I II I I II II I I II I II I II I I II I orf6-1 EDQNWKGLQEQFPAYAANFPVWADQANAMVQYAVWTTLAAVGVGANLQHYNPLPDAAIA 40 50 60 70 80 90 170 180 190 200 orf6a. pep KAWNIPENWLLRAQMVIGGIEGAAGEKTFEPVAERLKVFGAX I I II I I I II I II II I I I I II II I I I I I II I II II I I I I I I I I orf6-1 KAWNIPENWLLRAQMVIGGIEGAAGEKTFEPVAERLKVFGAX 100 110 120 130 Homology with a predicted ORF from N.gonorrhoeae ORF6 shows 95.7% identity over a 140aa overlap with a predicted ORF (ORF6ng) from N.gonorrhoeae: orf6. pep KVWQFVEXPLRAWPADSFEPTAQKLNLFK 30 II II I I I I I II I II I I I II I I II I : I I I orfδng SNVSLDMSNPTVLRMGLPLYIASLRRGAIYKVWQFVEDALRAWPADSFEPTAQKLKLFK 64 orf6.pep AGAATILFYEDQNWKGLQEQFPAYAANFPVWADQANAMVQYAVWTTLAAVGVGANLQHY 90 I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I : I I I I I I I orfδng AGAATILFYEDQNWKGLQEQFPAYAANFPVWADQANAMVQYAVWTTLAAVGAGANLQHY 124 orf6. pep NPLPDAAIAKAWNIPENWLLRAQMVIGGIEGAAGEKTFEPVAERLKVFGA 140 I I I I I : I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I : I I I I I I I I I I I I I orfδng NPLPDVAIAKAWNIPENWLLRAQMVIGGIEGAAGEKVFEPVAERLKVFGA 174 The co implete length ORFόng nucleotide sequence <SEQ ID 661> was identified as: 1 ATGGCCGTTG CGTCAAATGT CAGCTTGGAT ATGTCCAATC CTACGGTGTT 51 ACGCATGGGA TTACCCTTAT ATATTGCGTC CCTAAGAAGG GGCGCAATAT 101 ATAAGGTGTG GCAATTTGTC GAAGACGCGC TGCGTGCCGT CGTGCCTGCC 151 GACAGTTTTG AACCGACCGC GCAAAAATTG AAGCTGTTTA AGGCGGGCGC 201 GGCAACCATT TTGTTTTATG AAGATCAAAA TGTCGTCAAA GGTTTGCAGG 251 AGCAGTTCCC TGCTTATGCC GCCAACTTTC CCGTTTGGGC GGACCAGGCG 301 AACGCTATGG TACAGTATGC CGTCTGGACG ACACTTGCCG CGGTCGGTGC 351 AGGTGCAAAT CTGCAACATT ACAACCCCTT GCCCGATGTG GCGATTGCTA 401 AAGCGTGGAA TATTCCCGAA AACTGGCTGT TGCGCGCGCA AATGGTTATC 451 GGTGGTATTG AAGGGGcggc aggtgaaaaa gtctttgaac CCGTTGCgga 501 acgtttgAAA GTGTTCGGCG CATAA This encodes a protein having amino acid sequence <SEQ ID 662>: 1 MAVASNVSLD MSNPTVLRMG LPLYIASLRR GAIYKVWQFV EDALRAWPA 51 DSFEPTAQKL KLFKAGAATI LFYEDQNWK GLQEQFPAYA ANFPVWADQA 101 NAMVQYAVWT TLAAVGAGAN LQHYNPLPDV AIAKAWNIPE NWLLRAQMVI 151 GGIEGAAGEK VFEPVAERLK VFGA* ORFόng and ORF6-1 show 96.9% identity in 131 aa overlap: 10 20 30 orf 6-1 . pep LRAWPADSFEPTAQKLNLFKAGAATILFY I I I I I I I I I I I I I I I I I : I I I I I I I I I I I I orf 6ng PTVLRMGLPLYIASLRRGAIYKVWQFVEDALRAWPADSFEPTAQKLKLFKAGAATILFY 20 30 40 50 60 70 40 50 60 70 80 90 orf 6-1 . pep EDQNWKGLQEQFPAYAANFPVWADQANAMVQYAVWTTLAAVGVGANLQHYNPLPDAAIA I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I : I I I I I I I I I I I M I I I orf δng EDQNWKGLQEQFPAYAANFPVWADQANAMVQYAVWTTLAAVGAGANLQHYNPLPDVAIA 80 90 100 110 120 130 100 110 120 130 orf 6-1 . pep KAWNIPENWLLRAQMVIGGIEGAAGEKTFEPVAERLKVFGAX I I I I I I I I I I I I I I I I I I I I I I I I I I M M I I I I I I I I I I I I orf δng KAWNIPENWLLRAQMVIGGIEGAAGEKVFEPVAERLKVFGAX 140 150 160 170 It is predicted that the proteins from N. meningitidis .and N.gonorrhoeae, and their epitopes, could be useful antigens for vaccines or diagnostics, or for raising antibodies. Example 79 The following partial DNA sequence was identified in N. meningitidis <SEQ ID 663> 1 . GGCTACAACT ACCTGTTCGC GCGCGGCAGC CGCATCGCCA ACTACCAAAT 51 CAACGGCATC CCCGTTGCCG ACGCGCTGGC CGATACGGGt CAATGCCAAC 101 ACCGCCGCCT ATGAGCGCGT AGAAGTCGTG CGCGGCGTGG CGGGGCTGCT 151 GGACGGCACG GGCGAGCCTT CCGCCACCGT CAATCTGGTG CGCAAACGCC 201 TGACCCGCAA GCCATTGTTT GAAGTCCGCG CCGAAGCgGG CAACCGcAAA 251 CATTTCGGGC TGGACGCGGA CGTATCGGGC AGCCTGAACA CCGAAG.crC 301 rCTGCGCgGC CGCCTGGTTT CCAcCTTCGG ACGCGGCGAC TCGTGGCGGC 351 GGCGCGAACG CAGCCGskAT GCCGAACTCT ACGGCATTTT GGAATACGAC 401 ATCGCACCGC AAACCCGCGT CCACGCArGC ATGGACTACC AGCAGGCGAA 451 AGAAACCGCC GACGCGCCGC TCAGcTACGC CGTGTACGAC AGCCAAGGTT 501 ATGCCACCGC CTTCGGCCCG AAAGACAACC CCGCCACAAA TTGGGCGAAC 551 AGCCACCACC GTGCGCTCAA CCTGTTCGCC GGCATCGAAC ACCGCTTCAA 601 CCAAGACTGG AAACTCAAAG CCGAATACGA CTAC.. This coπesponds to the .amino acid sequence <SEQ ID 664; ORF23>: 1 ..GYNYLFARGS RIANYQINGI PVADALADTG NANTAAYERV EWRGVAGLL 51 DGTGEPSATV NLVRKRLTRK PLFEVRAEAG NRKHFGLDAD VSGSLNTEXX 101 LRGRLVSTFG RGDSWRRRER SRXAELYGIL EYDIAPQTRV HAXMDYQQAK 151 ETADAPLSYA VYDSQGYATA FGPKDNPATN WANSHHRALN LFAGIEHRFN 201 QDWKLKAEYD Y.. Further work revealed the complete nucleotide sequence <SEQ ID 665>: 1 ATGACACGCT TCAAATATTC CCTGCTGTTT GCCGCCCTGT TGCCCGTGTA 51 CGCGCAGGCC GATGTTTCTG TTTCAGACGA CCCCAAACCG CAGGAAAGCA 101 CTGAATTGCC GACCATCACC GTTACCGCCG ACCGCACCGC GAGTTCCAAC 151 GACGGCTACA CTGTTTCCGG CACGCACACC CCGCTCGGGC TGCCCATGAC 201 CCTGCGCGAA ATCCCGCAGA GCGTCAGCGT CATCACATCG CAACAAATGC 251 GCGACCAAAA CATCAAAACG CTCGACCGCG CCCTGTTGCA GGCGACCGGC 301 ACCAGCCGCC AGATTTACGG CTCCGACCGC GCGGGCTACA ACTACCTGTT 351 CGCGCGCGGC AGCCGCATCG CCAACTACCA AATCAACGGC ATCCCCGTTG 401 CCGACGCGCT GGCCGATACG GGCAATGCCA ACACCGCCGC CTATGAGCGC 451 GTAGAAGTCG TGCGCGGCGT GGCGGGGCTG CTGGACGGCA CGGGCGAGCC 501 TTCCGCCACC GTCAATCTGG TGCGCAAACG CCTGACCCGC AAGCCATTGT 551 TTGAAGTCCG CGCCGAAGCG GGCAACCGCA AACATTTCGG GCTGGACGCG 601 GACGTATCGG GCAGCCTGAA CACCGAAGGC ACGCTGCGCG GCCGCCTGGT 651 TTCCACCTTC GGACGCGGCG ACTCGTGGCG GCGGCGCGAA CGCAGCCGCG 701 ATGCCGAACT CTACGGCATT TTGGAATACG ACATCGCACC GCAAACCCGC 751 GTCCACGCAG GCATGGACTA CCAGCAGGCG AAAGAAACCG CCGACGCGCC 801 GCTCAGCTAC GCCGTGTACG ACAGCCAAGG TTATGCCACC GCCTTCGGCC 851 CGAAAGACAA CCCCGCCACA AATTGGGCGA ACAGCCGCCA CCGTGCGCTC 901 AACCTGTTCG CCGGCATCGA ACACCGCTTC AACCAAGACT GGAAACTCAA 951 AGCCGAATAC GACTACACCC GCAGCCGCTT CCGCCAGCCC TACGGCGTAG 1001 CAGGCGTGCT TTCCATCGAC CACAACACCG CCGCCACCGA CCTGATTCCC 1051 GGTTATTGGC ACGCCGACCC GCGCACCCAC AGCGCCAGCG TGTCATTGAT 1101 CGGCAAATAC CGCCTGTTCG GCCGCGAACA CGATTTAATC GCGGGTATCA 1151 ACGGTTACAA ATACGCCAGC AACAAATACG GCGAACGCAG CATCATCCCC 1201 AACGCCATTC CCAACGCCTA CGAATTTTCC CGCACGGGTG CCTACCCGCA 1251 GCCTGCATCG TTTGCCCAAA CCATCCCGCA ATACGGCACC AGGCGGCAAA 1301 TCGGCGGCTA TCTCGCCACC CGTTTCCGCG CCGCCGACAA CCTTTCGCTG 1351 ATTTTGGGCG GACGATACAC CCGTTACCGC ACCGGCAGCT ACGACAGCCG 1401 CACACAAGGC ATGACCTATG TGTCCGCCAA CCGTTTCACC CCCTACACAG 1451 GCATCGTGTT CGACCTGACC GGCAACCTGT CTCTTTACGG CTCGTACAGC 1501 AGCCTGTTCG TCCCGCAATC GCAAAAAGAC GAACACGGCA GCTACCTGAA 1551 ACCCGTAACC GGCAACAATC TGGAAGCCGG CATCAAAGGC GAATGGCTTG 1601 AAGGCCGTCT GAACGCATCC GCCGCCGTGT ACCGCGCCCG TAAAAACAAC 1651 CTCGCCACCG CAGCAGGACG CGACCCGAGC GGCAACACCT ACTACCGCGC 1701 CGCCAACCAA GCCAAAACCC ACGGCTGGGA AATCGAAGTC GGCGGCCGCA 1751 TCACGCCCGA ATGGCAGATA CAGGCAGGTT ACAGCCAAAG CAAAACCCGC 1801 GACCAAGACG GCAGCCGCCT GAACCCCGAC AGCGTACCCG AACGCAGCTT 1851 CAAACTCTTC ACTGCCTACC ACTTTGCCCC CGAAGCCCCC AGCGGCTGGA 1901 CCATCGGCGC AGGCGTGCGC TGGCAGAGCG AAACCCACAC CGACCCTGCC 1951 ACGCTCCGCA TCCCCAACCC CGCCGCCAAA GCCCGCGCCG CCGACAACAG 2001 CCGCCAAAAA GCCTACGCCG TCGCCGACAT CATGGCGCGT TACCGCTTCA 2051 ATCCGCGCGC CGAACTGTCG CTGAACGTGG ACAATCTGTT CAACAAACAC 2101 TACCGCACCC AGCCCGACCG CCACAGCTAC GGCGCACTGC GGACAGTGAA 2151 CGCGGCGTTT ACCTATCGGT TTAAATAA This coπesponds to the amino acid sequence <SEQ LD 666; ORF23-1 > • 1 MTRFKYSLLF AALLPVYAQA DVSVSDDPKP QESTELPTIT VTADRTASSN 51 DGYTVSGTHT PLGLPMTLRE IPQSVSVITS QQMRDQNIKT LDRALLQATG 101 TSRQIYGSDR AGYNYLFARG SRIANYQING IPVADALADT GNANTAAYER 151 VEWRGVAGL LDGTGEPSAT VNLVRKRLTR KPLFEVRAEA GNRKHFGLDA 201 DVSGSLNTEG TLRGRLVSTF GRGDSWRRRE RSRDAELYGI LEYDIAPQTR 251 VHAGMDYQQA KETADAPLSY AVYDSQGYAT AFGPKDNPAT NWANSRHRAL 301 NLFAGIEHRF NQDWKLKAEY DYTRSRFRQP YGVAGVLSID HNTAATDLIP 351 GYWHADPRTH SASVSLIGKY RLFGREHDLI AGINGYKYAS NKYGERSIIP 401 NAIPNAYEFS RTGAYPQPAS FAQTIPQYGT RRQIGGYLAT RFRAADNLSL 451 ILGGRYTRYR TGSYDSRTQG MTYVSANRFT PYTGIVFDLT GNLSLYGSYS 501 SLFVPQSQKD EHGSYLKPVT GNNLEAGIKG EWLEGRLNAS AAVYRARKNN 551 LATAAGRDPS GNTYYRAANQ AKTHGWEIEV GGRITPEWQI QAGYSQSKTR 601 DQDGSRLNPD SVPERSFKLF TAYHFAPEAP SGWTIGAGVR WQSETHTDPA 651 TLRIPNPAAK ARAADNSRQK AYAVADIMAR YRFNPRAELS LNVDNLFNKH 701 YRTQPDRHSY GALRTVNAAF TYRFK* Computer analysis of this amino acid sequence gave the following results: Homology with the femc-pseudobactin receptor PupB of Pseudomonas putida (accession number P38047) ORF23 and PupB protein show 32% aa identity in 205aa overlap: Orf23 6 FARGSRIANYQINGIPVADALADTGNANTAAYERVEWRGVAGLLDGTGEPSATVNLVRK 65 ++RG I NY+++G+P + L D + + A ++RVE+VRG GL+ G G PSAT+NL+RK PupB 215 WSRGFAIQNYEVDGVPTSTRL-DNYSQSMAMFDRVEIVRGATGLISGMGNPSATINLIRK 273 Orf23 66 RLTRKPLFEVRAEAGNRKHFGLDADVSGSLNTEXXLRGRLVSTFXXXXXXXXXXXXXXAE 125 R T + + EAGN +G DVSG L +RGR V+ + PupB 274 RPTAEAQASITGEAGNWDRYGTGFDVSGPLTETGNIRGRFVADYKTEKAWIDRYNQQSQL 333 Orf23 126 LYGILEYDIAPQTRVHAXMDYQQAKETADAPLSYAVYD—SQGYATAFGPKDNPATNWAN 183 +YGI E+D++ T + Y + D+PL + S G T N A +W+ PupB 334 MYGITEFDLSEDTLLTVGFSY—LRSDIDSPLRSGLPTRFSTGERTNLKRSLNAAPDWSY 391 Orf23 184 SHHRALNLFAGIEHRFNQDWKLKAE 208 + H + F IE + W K E PupB 392 NDHEQTSFFTSIEQQLGNGWSGKIE 416 Homology with a predicted ORF from N.meninsitidis (strain Al ORF23 shows 95.7% identity over a 21 laa overlap with an ORF (ORF23a) from sfrain A of N meningitidis: 10 20 30 orf23.pep GYΝYLFARGSRIAΝYQIΝGIPVADALADTG I II I II II I I II I I II I I II I II I I I I I I I orf23a QMRDQΝIKALDRALLQATGTSRQIYGSDRAGYΝYLFARGSRIAΝYQIΝGIPVADALADTG 90 100 110 120 130 140 40 50 60 70 80 90 orf23.pep ΝAΝTAAYERVEWRGVAGLLDGTGEPSATVΝLVRKRLTRKPLFEVRAEAGΝRKHFGLDAD II I I II II I I I II I I I I II I II I II I I I I I I I I I I I I I I II I I I I I I I II II I I I I II orf23a ΝAΝTAAYERVEWRGVAGLLDGTGEPSATVΝLVRKRPTRKPLFEVRAEAGΝRKHFGLGAD 150 160 170 180 190 200 100 110 120 130 140 150 orf23. pep VSGSLΝTEXXLRGRLVSTFGRGDSWRRRERSRXAELYGILEYDIAPQTRVHAXMDYQQAK I I II I M I : II I I I I I I I I I I II I I : II I II II I I I I I I I I II II I I II I I II II II orf23a VSGSLΝAEGTLRGRLVΞTFGRGDSWRQRERSRDAELYGILEYDIAPQTRVHAGMDYQQAK 210 220 230 240 250 260 160 170 180 190 200 210 orf23. pep ETADAPLSYAVYDSQGYATAFGPKDΝPATΝWAΝSHHRALΝLFAGIEHRFΝQDWKLKAEYD I II I I I II I I I II II I II I I I I II II I II I II I I : I I I I I I I I II II I I II II I II II II orf23a ETADAPLSYAVYDSQGYATAFGPKDΝPATΝWAΝSRHRALΝLFAGIEHRFΝQDWKLKAEYD 270 280 290 300 310 320 orf23. pep Y orf23a YTRSRFRQPYGVAGVLSIDHΝTAATDLIPGYWHADPRTHSASVSLIGKYRLFGREHDLIA 330 340 350 360 370 380 The complete length ORF23a nucleotide sequence <SEQ ID 667> is: 1 ATGACACGCT TCAAATATTC CCTGCTGTTT GCCGCCCTGT TGCCCGTGTA 51 CGCGCAGGCC GATGTTTCTG TTTCAGACGA CCCAAAACCG CAGGAAAGCA 101 CTGAATTGCC GACCATCACC GTTACCGCCG ACCGCACCGC GAGTTCCAAC 151 GACGGCTACA CTGTTTCCGG CACGCACACC CCGCTCGGGC TGCCCATGAC 201 CCTGCGCGAA ATCCCGCAGA GCGTCAGCGT CATCACATCG CAACAAATGC 251 GCGACCAAAA CATCAAAGCG CTCGACCGCG CCCTGTTGCA GGCGACCGGC 301 ACCAGCCGCC AGATTTACGG CTCCGACCGC GCGGGCTACA ACTACCTGTT 351 CGCGCGCGGC AGCCGCATCG CCAACTACCA AATCAACGGC ATCCCCGTTG 401 CCGACGCGCT GGCCGATACG GGCAATGCCA ACACCGCCGC CTATGAGCGC 451 GTAGAAGTCG TGCGCGGCGT GGCGGGGCTG CTGGACGGCA CGGGCGAGCC 501 TTCCGCCACC GTCAATCTGG TGCGCAAACG CCCGACCCGC AAGCCATTGT 551 TTGAAGTCCG CGCCGAAGCG GGCAACCGCA AACATTTCGG GCTGGGCGCG 601 GACGTATCGG GCAGCCTGAA TGCCGAAGGC ACGCTGCGCG GCCGCCTGGT 651 TTCCACCTTC GGACGCGGCG ACTCGTGGCG GCAGCGCGAA CGCAGCCGCG 701 ATGCCGAACT CTACGGCATT TTGGAATACG ACATCGCACC GCAAACCCGC 751 GTCCACGCAG GCATGGACTA CCAGCAGGCG AAAGAAACCG CCGACGCGCC 801 GCTCAGCTAC GCCGTGTACG ACAGCCAAGG TTATGCCACC GCCTTCGGCC 851 CGAAAGACAA CCCCGCCACA AATTGGGCGA ACAGCCGCCA CCGTGCGCTC 901 AACCTGTTCG CCGGCATCGA ACACCGCTTC AACCAAGACT GGAAACTCAA 951 AGCCGAATAC GACTACACCC GCAGCCGCTT CCGCCAGCCC TACGGCGTAG 1001 CAGGCGTGCT TTCCATCGAC CACAACACCG CCGCCACCGA CCTGATTCCC 1051 GGTTATTGGC ACGCCGACCC GCGCACCCAC AGCGCCAGCG TGTCATTAAT 1101 CGGCAAATAC CGCCTGTTCG GCCGCGAACA CGATTTAATC GCGGGTATCA 1151 ACGGTTACAA ATACGCCAGC AACAAATACG GCGAACGCAG CATCATCCCC 1201 AACGCCATTC CCAACGCCTA CGAATTTTCC CGCACGGGTG CCTACCCGCA 1251 GCCTGCATCG TTTGCCCAAA CCATCCCGCA ATACGGCACC AGGCGGCAAA 1301 TCGGCGGCTA TCTCGCCACC CGTTTCCGCG CCGCCGACAA CCTTTCGCTG 1351 ATACTCGGCG GCAGATACAG CCGTTACCGC ACCGGCAGCT ACGACAGCCG 1401 CACACAAGGC ATGACCTATG TGTCCGCCAA CCGTTTCACC CCCTACACAG 1451 GCATCGTGTT CGACCTGACC GGCAACCTGT CGCTTTACGG CTCGTACAGC 1501 AGCCTGTTCG TCCCGCAATC GCAAAAAGAC GAACACGGCA GCTACCTGAA 1551 ACCCGTAACC GGCAACAATC TGGAAGCCGG CATCAAAGGC GAATGGCTTG 1601 AAGGCCGTCT GAACGCATCC GCCGCCGTGT ACCGCGCCCG TAAAAACAAC 1651 CTCGCCACCG CAGCAGGACG CGACCCGAGC GGCAACACCT ACTACCGCGC 1701 CGCCAACCAA GCCAAAACCC ACGGCTGGGA AATCGAAGTC GGCGGCCGCA 1751 TCACGCCCGA ATGGCAGATA CAGGCAGGTT ACAGCCAAAG CAAAACCCGC 1801 GACCAAGACG GCAGCCGCCT GAACCCCGAC AGCGTACCCG AACGCAGCTT 1851 CAAACTCTTC ACTGCCTACC ACTTTGCCCC CGAAGCCCCC AGCGGCTGGA 1901 CCATCGGCGC AGGCGTGCGC TGGCAGAGCG AAACCCACAC CGACCCTGCC 1951 ACGCTCCGCA TCCCCAACCC CGCCGCCAAA GCCCGCGCCG CCGACAACAG 2001 CCGCCAAAAA GCCTACGCCG TCGCCGACAT CATGGCGCGT TACCGCTTCA 2051 ATCCGCGCGC CGAACTGTCG CTGAACGTGG ACAATCTGTT CAACAAACAC 2101 TACCGCACCC AGCCCGACCG CCACAGCTAC GGCGCACTGC GGACAGTGAA 2151 CGCGGCGTTT ACCTATCGGT TTAAATAA This encodes a protein having amino acid sequence <SEQ ED 668>: 1 MTRFKYSLLF AALLPVYAQA DVSVSDDPKP QESTELPTIT VTADRTASSN 51 DGYTVSGTHT PLGLPMTLRE IPQSVSVITS QQMRDQNIKA LDRALLQATG 101 TSRQIYGSDR AGYNYLFARG SRIANYQING IPVADALADT GNANTAAYER 151 VEWRGVAGL LDGTGEPSAT VNLVRKRPTR KPLFEVRAEA GNRKHFGLGA 201 DVSGSLNAEG TLRGRLVSTF GRGDSWRQRE RSRDAELYGI LEYDIAPQTR 251 VHAGMDYQQA KETADAPLSY AVYDSQGYAT AFGPKDNPAT NWANSRHRAL 301 NLFAGIEHRF NQDWKLKAEY DYTRSRFRQP YGVAGVLSID HNTAATDLIP 351 GYWHADPRTH SASVSLIGKY RLFGREHDLI AGINGYKYAS NKYGERSIIP 401 NAIPNAYEFS RTGAYPQPAS FAQTIPQYGT RRQIGGYLAT RFRAADNLSL 451 ILGGRYSRYR TGΞYDSRTQG MTYVSANRFT PYTGIVFDLT GNLSLYGSYS 501 SLFVPQSQKD EHGSYLKPVT GNNLEAGIKG EWLEGRLNAS AAVYRARKNN 551 LATAAGRDPS GNTYYRAANQ AKTHGWEIEV GGRITPEWQI QAGYSQSKTR 601 DQDGSRLNPD SVPERSFKLF TAYHFAPEAP SGWTIGAGVR WQSETHTDPA 651 TLRIPNPAAK ARAADNSRQK AYAVADIMAR YRFNPRAELS LNVDNLFNKH 701 YRTQPDRHSY GALRTVNAAF TYRFK* ORF23a and ORF23-1 show 99.2% identity in 725 aa overlap: 10 20 30 40 50 60 orf23a . pep MTRFKYSLLFAALLPVYAQADVSVSDDPKPQESTELPTITVTADRTASSNDGYTVSGTHT I I I I I I II I I I I II I I II I I I I I I II I II I I I I I II I I II I II II I I I II I MMM I I I orf23-1 MTRFKYSLLFAALLPVYAQADVSVSDDPKPQESTELPTITVTADRTASSNDGYTVSGTHT 10 20 30 40 50 60 70 80 90 100 110 120 orf23a . pep PLGLPMTLREIPQSVSVITSQQMRDQNIKALDRALLQATGTSRQIYGSDRAGYNYLFARG I I I I I I II I II II II I II I II I I I I I I I M II II I I II I I I I I I II I I I I II I I I I II I I orf 23-1 PLGLPMTLREIPQSVSVITSQQMRDQNIKTLDRALLQATGTSRQIYGSDRAGYNYLFARG 70 80 90 100 110 120 130 140 150 160 170 180 orf 23a . pep SRIANYQINGIPVADALADTGNANTAAYERVEWRGVAGLLDGTGEPSATVNLVRKRPTR I I I II II II I II I I I I II II I II I I II I I I I I I I I I I I II II I I II I I II I I II II I II orf23-1 SRIANYQINGIPVADALADTGNANTAAYERVEWRGVAGLLDGTGEPSATVNLVRKRLTR 130 140 150 160 170 180 190 200 210 220 230 240 orf23a . pep KPLFEVRAEAGNRKHFGLGADVSGSLNAEGTLRGRLVSTFGRGDSWRQRERSRDAELYGI II I I II II I I I I II II I I I I II II II: I I I I I I I I I I I I II I I II I : II I I II II I I II orf23-1 KPLFEVRAEAGNRKHFGLDADVSGSLNTEGTLRGRLVSTFGRGDSWRRRERSRDAELYGI 190 200 210 220 230 240 250 260 270 280 290 300 orf23a . pep LEYDIAPQTRVHAGMDYQQAKETADAPLSYAVYDSQGYATAFGPKDNPATNWANSRHRAL I II II I I I I I I I I I I II I II I I I II II I I I I I I I I I I I I I I I I I I I I II II I I I I I II I I orf23-1 LEYDIAPQTRVHAGMDYQQAKETADAPLSYAVYDSQGYATAFGPKDNPATNWANSRHRAL 250 260 270 280 290 300 310 320 330 340 350 360 orf23a . pep NLFAGIEHRFNQDWKLKAEYDYTRSRFRQPYGVAGVLSIDHNTAATDLIPGYWHADPRTH II I II II II I II I II I I II I II II I II I I I I I I I I I I I I I II I I II I I I II I I I II I I I I orf23-1 NLFAGIEHRFNQDWKLKAEYDYTRSRFRQPYGVAGVLSIDHNTAATDLIPGYWHADPRTH 310 320 330 340 350 360 370 380 390 400 410 420 orf23a . pep SASVSLIGKYRLFGREHDLIAGINGYKYASNKYGERSIIPNAIPNAYEFSRTGAYPQPAS II I I I I I II I I II II II I I I I I I II II I I I II I I I I I II I I II II I II I I I I I I I I I II I orf23-1 SASVSLIGKYRLFGREHDLIAGINGYKYASNKYGERSIIPNAIPNAYEFSRTGAYPQPAS 370 380 390 400 410 420 430 440 450 460 470 480 orf23a . pep FAQTIPQYGTRRQIGGYLATRFRAADNLSLILGGRYSRYRTGSYDSRTQGMTYVSANRFT I II II I I I I I I I II I I II I II I II II II I I II I I I I : I I I I I I I I I I I I II I I I I II I I I orf23-1 FAQTIPQYGTRRQIGGYLATRFRAADNLSLILGGRYTRYRTGSYDSRTQGMTYVSANRFT 430 440 450 460 470 480 490 500 510 520 530 540 orf23a. pep PYTGIVFDLTGNLSLYGSYSSLFVPQSQKDEHGSYLKPVTGNNLEAGIKGEWLEGRLNAS I I II II I II I II I II II II I I II II I II II I I II II II II I I II II I I II I I I I II I I I I orf23-1 PYTGIVFDLTGNLSLYGSYSSLFVPQSQKDEHGSYLKPVTGNNLEAGIKGEWLEGRLNAS 490 500 510 520 530 540 550 560 570 580 590 600 orf23a. pep AAVYRARKNNLATAAGRDPSGNTYYRAANQAKTHGWEIEVGGRITPEWQIQAGYSQSKTR I I I I I I I II II II I I I II II I II I II II I I I I I I I I I II I I I I I I I I I I II I I I I I II I I orf23-1 AAVYRARKNNLATAAGRDPSGNTYYRAANQAKTHGWEIEVGGRITPEWQIQAGYSQSKTR 550 560 570 580 590 600 610 620 630 640 650 660 orf23a. pep DQDGSRLNPDSVPERSFKLFTAYHFAPEAPSGWTIGAGVRWQSETHTDPATLRIPNPAAK II I I I I I I I I I I I I I I I II I II I II I I I I I I II I I I II II I I II I I I I II I I II II I I II orf23-1 DQDGSRLNPDSVPERSFKLFTAYHFAPEAPSGWTIGAGVRWQSETHTDPATLRIPNPAAK 610 620 630 640 650 660 670 680 690 700 710 720 orf23a . pep ARAADNSRQKAYAVADIMARYRFNPRAELSLNVDNLFNKHYRTQPDRHSYGALRTVNAAF II I II I I I I I I I II I I I II I I I I I I I II I II I II I I I I I I I I I I I I I I I II I I I I I II I I orf23-1 ARAADNSRQKAYAVADIMARYRFNPRAELSLNVDNLFNKHYRTQPDRHSYGALRTVNAAF 670 680 690 700 710 720 orf23a. pep TYRFKX II I I I I orf23-l TYRFKX Homology with a predicted ORF from N.sonorrhoeae ORF23 shows 93.4% identity over a 21 laa overlap with a predicted ORF (ORF23.ng) from N gonorrhoeae: orf23.pep GYΝYLFARGSRIAΝYQIΝGIPVADALADTGΝAΝTAAYERVE RGVAGLLD 51 I II I II II II I II I II I II II II I II I I I I II I II II I I I I II II I I I I I orf23ng SAVDACRIPGYΝYLFARGSRIAΝYQIΝGIPVADALADTGΝAΝTAAYERVEWRGVAGLPD 60 orf23.pep GTGEPSATVΝLVRKRLTRKPLFEVRAEAGΝRKHFGLDADVSGSLΝTEXXLRGRLVSTFGR 111 II I I II II I I II I I : I I I I II I I II I I I I I II I I I I I I II I II : I : I I I I I I II II I orf23ng GTGEPSATVΝLVRKHPTRKPLFEVRAEAGΝRKHFGLGADVSGSLΝAEGTLRGRLVSTFGR 120 orf23.pep GDSWRRRERSRXAELYGILEYDIAPQTRVHAXMDYQQAKETADAPLSYAVYDSQGYATAF 171 Mill: I I I I I I I II II I II I I I I I II II I II II I I I I I I I I I I I I I I II I I I I I II orf23ng GDSWRQLERSRDAELYGILEYDIAPQTRVHAGMDYQQAKETADAPLSYAVYDSQGYATAF 180 orf23.pep GPKDΝPATΝWAΝSHHRALΝLFAGIEHRFΝQDWKLKAEYDY 211 I I II I II I I I : I I :: II II I II I II I I I I II II II I I I I I orf23ng GPKDΝPATΝWSΝSRΝRALΝLFAGIEHRFΝQDWKLKAEYDYTRSRFRQPYGVAGVLSIDHS 240 The ORF23ng nucleotide sequence <SEQ ID 669> is predicted to encode a protein comprising amino acid sequence <SEQ ID 670>: 1 SAVDACRIPG YΝYLFARGSR IAΝYQIΝGIP VADALADTGΝ AΝTAAYERVE 51 WRGVAGLPD GTGEPSATVΝ LVRKHPTRKP LFEVRAEAGΝ RKHFGLGADV 101 SGSLΝAEGTL RGRLVSTFGR GDSWRQLERS RDAELYGILE YDIAPQTRVH 151 AGMDYQQAKE TADAPLSYAV YDSQGYATAF GPKDΝPATΝW SΝSRΝRALΝL 201 FAGIEHRFΝQ DWKLKAEYDY TRSRFRQPYG VAGVLSIDHS TAATDLIPGY 251 WHADPRTHSA SMSLTGKYRL FGREHDLIAG IΝGYKYASΝK YGERSIIPΝA 301 IPΝAYEFSRT GAYPQPSSFA QTIPQYDTRR QIGGYLATRF RAADΝLSLIL 351 GGRYSRYRAG SYΝSRTQGMT YVSAΝRFTPY TGIVFDLTGΝ LSLYGSYSSL 401 FVPQLQKDEH GSYLKPVTGΝ ΝLEADIKGEW LEGRLΝASAA VYRARKΝΝLA 451 TAAGRDQSGΝ TYYRAAΝQAK THGWEIEVGG RITPEWQIQA GYSQSKPRDQ 501 DGSRLΝPDSV PERSFKLFTA YHLAPEAPSG RTIGAGVRRQ GETHTDPAAL 551 RIPΝPAAKAR AVAΝSRQKAY AVADIMARYR FΝPRTELSLΝ VDΝLFΝKHYR 601 TQPDRHSYGA LRTVΝAAFTY RFK* Further work revealed the complete nucleotide sequence <SEQ ID 671>: 1 ATGACACGCT TCAAATACTC CCTGCTTTTT GCCGCCCTGC TACCCGTGTA 51 CGCGCAGGCC GATGTTTCTG TTTCAGACGA CCCCAAACCG CAGGAAAGCA 101 CCGAATTGCC GACCATCACC GTTACCGCCG ACCGCACCGC GAGTTCCAAC 151 GACGGCTACA CCGTTTCCGG CACGCACACC CCGTTCGGGC TGCCCATGAC 201 CCTGCGCGAA ATCCCGCAGA GCGTCAGCGT CATCACATCG CAACAAATGC 251 GCGACCAAAA CATCAAAACG CTCGACCGCG CCCTGTTGCA GGCGACCGGC 301 ACCAGCCGCC AGATTTACGG CTCCGACCGC GCGGGCTACA ACTACCTGTT 351 CGCGCGCGGC AGCCGCATCG CCAACTACCA AATCAACGGC ATCCCCGTTG 401 CCGACGCGCT GGCCGATACG GGCAATGCCA ACACCGCCGC CTATGAGCGC 451 GTAGAAGTCG TGCGCGGCGT GGCGGGGCTG CCGGACGGCA CGGGCGAGCC 501 TTCTGCCACC GTCAATCTGG TACGCAAACA CCCGACCCGC AAGCCATTGT 551 TTGAAGTCCG CGCCGAAGCC GGCAACCGCA AACATTTCGG GCTGGGCGCG 601 GACGTATCGG GCAGCCTGAA CGCCGAAGGC ACGCTGCGCG GCCGCCTGGT 651 TTCCACCTTC GGACGCGGCG ACTCGTGGCG GCAGCTCGAA CGCAGCCGCG 701 ATGCCGAACT CTACGGCATT TTGGAATACG ACATCGCACC GCAAACCCGC 751 GTCCACGCAG GCATGGACTA CCAGCAGGCG AAAGAAACCG CAGACGCGCC 801 GCTCAGCTAC GCCGTGTACG ACAGCCAAGG TTATGCCACC GCCTTCGGCC 851 CAAAAGACAA CCCCGCCACA AATTGGTCGA ACAGCCGCAA CCGTGCGCTC 901 AACCTGTTCG CCGGCATAGA ACACCGCTTC AACCAAGACT GGAAACTCAA 951 AGCCGAATAC GACTACACCC GTAGCCGCTT CCGCCAGCCC TACGGTGTGG 1001 CAGGCGTACT TTCCATCGAC CACAGCACTG CCGCCACCGA CCTGATTCCC 1051 GGTTATTGGC ACGCcgatcc GCGCACCCAC AGCGCCAGCA TGTCATTGAC 1101 CGGCAAATAC CgcctGTTCG GCCGCGAGCA CGATTTAATC GCGGGTATCA 1151 ACGGCTACAA ATACGCCAGC AACAAATACG GCGAACGCAG CATCATTCCC 1201 AACGCCATTC CCAACGCCTA CGAATTTTCC CGCACGGGCG CCTATCCGCA 1251 GCCATCATCG TTTGCCCAAA CCATCCCGCA ATACGACACC AGGCGGCAAA 1301 TCGGCGGCTA TCTCGCCACC CGTTTCCGCG CCGCCGACAA CCTTTCGCTG 1351 ATACTCGGCG GCAGATACAG CCGCTACCGC GCAGGCAGCT ACAACAGCCG 1401 CACACAAGGC ATGACCTATG TGTCCGCCAA CCGTTTCACC CCCTACACAG 1451 GCATCGTGTT CGATCTGACC GGCAACCTGT CGCTTTACGG CTCGTACAGC 1501 AGCCTGTTCG TCCCGCAATT GCAAAAAGAC GAACACGGCA GCTACCTGAA 1551 ACCCGTAACC GGCAACAATC TGGAAGCCGA CATCAAAGGC GAATGGCTTG 1601 AAGGGCGTCT GAACGCATCC GCCGCCGTGT ACCGCGCCCG TAAAAACAAC 1651 CTCGCCACCG CAGCAGGACG CGACCAGAGC GGCAACACCT ACTATCGCGC 1701 CGCCAACCAA GCCAAAACCC ACGGCTGGGA AATCGAAGTC GGCGGCCGCA 1751 TCACGCCCGA ATGGCAGATA CAGGCAGGCT ACAGCCAAAG CAAACCCCGC 1801 GACCAAGACG GCAGCCGCCT GAACCCCGAC AGCGTAcCCG AACGCAGCTT 1851 CAAACTCTTC ACCGCCTACC ACTTAGCCCC CGAAGCCCCC AGCGGCCGGA 1901 CCATcggTGC GGGTGTGCGC CGGCAGGGCG AAACCCACAC CGACCCAGCC 1951 GCGCTCCGCA TCCCCAACCC CGCCGCCAAA GCCCGCGCCG TCGCCAACAG 2001 CCGCCAGAAA GCCTACGCCG TCGCCGACAT CATGGCGCGT TACCGCTTCA 2051 ATCCGCGCAC CGAACTGTCG CTGAACGTGG ACAACCTGTT CAACAAACAC 2101 TACCGCACCC AGCCCGACCG CCACAGCTAC GGCGCACTGC GGACAGTGAA 2 Δ15D1 CGϋLCGijϋGC>-G4jTiT 1T 1 A rtυCCυTiArtTiCuuGυGTi T j.T iArtΛAArtTiArtAΛ This coπesponds to the .amino acid sequence <SEQ ID 672; ORF23ng-l>: 1 MTRFKYSLLF AALLPVYAQA DVSVSDDPKP QESTELPTIT VTADRTASSN 51 DGYTVSGTHT PFGLPMTLRE IPQSVSVITS QQMRDQNIKT LDRALLQATG 101 TSRQIYGSDR AGYNYLFARG SRIANYQING IPVADALADT GNANTAAYER 151 VEWRGVAGL PDGTGEPSAT VNLVRKHPTR KPLFEVRAEA GNRKHFGLGA 201 DVSGSLNAEG TLRGRLVSTF GRGDSWRQLE RSRDAELYGI LEYDIAPQTR 251 VHAGMDYQQA KETADAPLSY AVYDSQGYAT AFGPKDNPAT NWSNSRNRAL 301 NLFAGIEHRF NQDWKLKAEY DYTRSRFRQP YGVAGVLSID HSTAATDLIP 351 GYWHADPRTH SASMSLTGKY RLFGREHDLI AGINGYKYAS NKYGERSIIP 401 NAIPNAYEFS RTGAYPQPSS FAQTIPQYDT RRQIGGYLAT RFRAADNLSL 451 ILGGRYSRYR AGSYNSRTQG MTYVSANRFT PYTGIVFDLT GNLSLYGSYS 501 SLFVPQLQKD EHGSYLKPVT GNNLEADIKG EWLEGRLNAS AAVYRARKNN 551 LATAAGRDQS GNTYYRAANQ AKTHGWEIEV GGRITPEWQI QAGYSQSKPR 601 DQDGSRLNPD SVPERSFKLF TAYHLAPEAP SGRTIGAGVR RQGETHTDPA 651 ALRIPNPAAK ARAVANSRQK AYAVADIMAR YRFNPRTELS LNVDNLFNKH 701 YRTQPDRHSY GALRTVNAAF TYRFK* ORF23ng-l and ORF23-1 show 95.9% identity in 725 aa overlap: 10 20 30 40 50 60 orf23-1. pep MTRFKYSLLFAALLPVYAQADVSVSDDPKPQESTELPTITVTADRTASSNDGYTVSGTHT II I I I II I I I II I I I I I II I I I I I I I I I I II I I II I II I II II II I II I I II I I II I II I orf23ng-l MTRFKYSLLFAALLPVYAQADVSVSDDPKPQESTELPTITVTADRTASSNDGYTVSGTHT 10 20 30 40 50 60 70 80 90 100 110 120 orf23-1. pep PLGLPMTLREIPQSVSVITSQQMRDQNIKTLDRALLQATGTSRQIYGSDRAGYNYLFARG M I II II II I II II I I I II I I I I I II I II II I I II I II I I I I I II I II I I II I I I I I I I I orf23ng-l PFGLPMTLREIPQSVSVITSQQMRDQNIKTLDRALLQATGTSRQIYGSDRAGYNYLFARG 70 80 90 100 110 120 130 140 150 160 170 180 orf23-1. pep SRIANYQINGIPVADALADTGNANTAAYERVEWRGVAGLLDGTGEPSATVNLVRKRLTR I I II I I I I I II I I I I I II I I I I I I II I I I II I I II I I I I I I I II I II I I I I I I IM II orf23ng-l SRIANYQINGIPVADALADTGNANTAAYERVEWRGVAGLPDGTGEPSATVNLVRKHPTR 130 140 150 160 170 180 190 200 210 220 230 240 orf23-1. pep KPLFEVRAEAGNRKHFGLDADVSGSLNTEGTLRGRLVSTFGRGDSWRRRERSRDAELYGI I II II II I II I II II II I I I I I II I I : I II I I II I I I I II II II I I: I I I I I I I I I I I orf23ng-l KPLFEVRAEAGNRKHFGLGADVSGSLNAEGTLRGRLVSTFGRGDSWRQLERSRDAELYGI 190 200 210 220 230 240 250 260 270 280 290 300 orf23-1. pep LEYDIAPQTRVHAGMDYQQAKETADAPLSYAVYDSQGYATAFGPKDNPATNWANSRHRAL I I I I I I I I I I I I I I I I I I I I I I I I I I I II I II II I II II I I I II I I I I I II I: I I I: I I I orf23ng-l LEYDIAPQTRVHAGMDYQQAKETADAPLSYAVYDSQGYATAFGPKDNPATNWSNSRNRAL 250 260 270 280 290 300 310 320 330 340 350 360 orf23-1. pep NLFAGIEHRFNQDWKLKAEYDYTRSRFRQPYGVAGVLSIDHNTAATDLIPGYWHADPRTH I I I II II I I I II I II II I I I I I I II I II I I I I I I II II II I : I I I I I I II II I I II I I II orf23ng-l NLFAGIEHRFNQDWKLKAEYDYTRSRFRQPYGVAGVLSIDHSTAATDLIPGYWHADPRTH 310 320 330 340 350 360 370 380 390 400 410 420 orf23-1. pep SASVSLIGKYRLFGREHDLIAGINGYKYASNKYGERSIIPNAIPNAYEFSRTGAYPQPAS MMM I II II II II II I II I I I II I I II I I II II I I I I II I I I I II I I II I I I I II : I orf23ng-l SASMSLTGKYRLFGREHDLIAGINGYKYASNKYGERSIIPNAIPNAYEFSRTGAYPQPSS 370 380 390 400 410 420 430 440 450 460 470 480 orf23-1.pep FAQTIPQYGTRRQIGGYLATRFRAADNLSLILGGRYTRYRTGSYDSRTQGMTYVSANRFT II I I I II I I I I II I I II II I II II I I I I I I I II I I : I I I : I II : I I I I I II I I I I I II I orf23ng-l FAQTIPQYDTRRQIGGYLATRFRAADNLSLILGGRYSRYRAGSYNSRTQGMTYVSANRFT 430 440 450 460 470 480 490 500 510 520 530 540 orf23-1. pep PYTGIVFDLTGNLSLYGSYSSLFVPQSQKDEHGSYLKPVTGNNLEAGIKGEWLEGRLNAS I II I II II II II I II II II I II II I I II II II I I I I I II II I I I I I I I I I I I I I I II I orf23ng-l PYTGIVFDLTGNLSLYGSYSSLFVPQLQKDEHGSYLKPVTGNNLEADIKGEWLEGRLNAS 490 500 510 520 530 540 550 560 570 580 590 600 orf23-1. pep AAVYRARKNNLATAAGRDPSGNTYYRAANQAKTHGWEIEVGGRITPEWQIQAGYSQSKTR I I I II I I I I I II I I I I I I I II II I II II II II I I I I I I I II II I II I I I II I I I I I I I orf23ng-l AAVYRARKNNLATAAGRDQSGNTYYRAANQAKTHGWEIEVGGRITPEWQIQAGYSQSKPR 550 560 570 580 590 600 610 620 630 640 650 660 orf23-1. pep DQDGSRLNPDSVPERSFKLFTAYHFAPEAPSGWTIGAGVRWQSETHTDPATLRIPNPAAK I I II II I II I I I II II II I II I II : I II I I I I II II I II I : I I I II I I : I I I I I I II I or 23ng-l DQDGSRLNPDSVPERSFKLFTAYHLAPEAPSGRTIGAGVRRQGETHTDPAALRIPNPAAK 610 620 630 640 650 660 670 680 690 700 710 720 orf23-1. pep ARAADNSRQKAYAVADIMARYRFNPRAELSLNVDNLFNKHYRTQPDRHSYGALRTVNAAF M I M II I II II II I II I II I II : I I II II II II I II II II I I II II II I I I I I II I orf23ng-l ARAVANSRQKAYAVADIMARYRFNPRTELSLNVDNLFNKHYRTQPDRHSYGALRTVNAAF 670 680 690 700 710 720 orf23-1. pep TYRFKX I I I I I I orf23ng-l TYRFKX ition, ORF23ng-l shows significant homology with an OMP from E.coli: sp|P16869|FHUE_ECOLI OUTER-MEMBRANE RECEPTOR FOR FE ( III ) -COPROGEN, FE(III)- FERRIOXAMINE B AND FE (III) -RHODOTRULIC ACID PRECURSOR >gι 11651542 | gnl | PID | dl015403 (D90745) Outer membrane protein FhuE precursor [Escherichia coli] >gι|1651545|gnl|PID|dl015405 (D90746) Outer membrane protein FhuE precursor [Escherichia coli] >gι 11787344 (AE000210) outer-membrane receptor for Fe(III)- coprogen, Fe (III) -femoxamme B and Fe (III) -rhodotrulic acid precursor [Escherichia coli] Length = 729 Score = 332 bits (843), Expect = 3e-90 Identities = 228/717 (31%), Positives = 350/717 (48%), Gaps = 60/717 (8%) Query: 38 TITVTADRTASSN--DGYTVSGTHTPFGLPMTLREIPQSVSVITSQQMRDQNIKTLDRAL 95 T+ V TA + + Y+V+ T + MT R+IPQSV++++ Q+M DQ ++TL + Sbjct: 43 TVIVEGSATAPDDGENDYSVTSTSAGTKMQMTQRDIPQSVTIVSQQRMEDQQLQTLGEVM 102 Query: 96 LQATGTSRQIYGSDRAGYNYLFARGSRIANYQINGIP VADALADTGNANTAA 147 G S+ SDRA Y ++RG +1 NY ++GIP + DAL+D A Sbjct: 103 ENTLGISKSQADSDRALY YSRGFQIDNYMVDGIPTYFESRWNLGDALSDM AL 154 Query: 148 YERVEWRGVAGLPDGTGEPSATVNLVRKHPTRKPLF-EVRAEAGNRKHFGLGADVSGSL 206 +ERVEWRG GL GTG PSA +N+VRKH T + +V AE G+ AD+ L Sbjct: 155 FERVEWRGATGLMTGTGNPSAAINMVRKHATSREFKGDVSAEYGSWNKERYVADLQΞPL 214 Query: 207 NAEGTLRGRLVSTFGRGDSWRQLERSRDAELYGILEYDIAPQTRVHAGMDYQQAKETADA 266 +G +R R+V - DSW S GI++ D+ T + AG +YQ+ + Sbjct: 215 TEDGKIRARIVGGYQNNDSWLDRYNSEKTFFSGIVDADLGDLTTLSAGYEYQRIDVNSPT 274 Query: 267 PLSYAVYDSQGYATAFGPKDNPATNWSNSRNRALNLFAGIEHRFNQDWKLKAEYDYTRSR 326 +++ G + ++ + A +W+ + +F ++ +F W+ ++ Sbjct: 275 WGGLPRWNTDGSSNSYDRARSTAPDWAYNDKEINKVFMTLKQQFADTWQATLNATHSEVE 334 Query: 327 F—RQPYGVAGVLSIDHSTAA—TDLIPGY WHADPRTHSA-SMSLTGKYRLFG 374 F + Y A V D ++ PG+ W++ R A + G Y LFG Sbjct: 335 FDSKMMYVDAYVNKADGMLVGPYΞNYGPGFDYVGGTGWNSGKRKVDALDLFADGSYELFG 394 Query: 375 REHDLIAGINGYKYASNKYGER—SIIPNAIPNAYEFSRTGAYPQPSSFAQTIPQYDTRR 432 R+H+L+ G Y +N+Y +1 P+ I + Y F+ G +PQ Q++ Q DT Sbjct: 395 RQHNLMFG-GSYSKQNNRYFSSWANIFPDEIGSFYNFN—GNFPQTDWSPQSLAQDDTTH 451 Query: 433 QIGGYLATRFRAADNLSLILGGRYSRYRAGSYNSRTQGMTY-VSANRFTPYTGIVFDXXX 491 Y ATR AD L LILG RY+ +R + +TY + N TPY G+VFD Sbjct: 452 MKSLYAATRVTLADPLHLILGARYTNWRVDT LTYSMEKNHTTPYAGLVFDIND 504 Query: 492 XXXXXXXXXXXFVPQLQKDEHGSYLKPVTGNNLEADIKGEWLEGRLNASAAVYRARKNNL 551 F PQ +D G YL P+TGNN E +K +W+ RL + A++R ++N+ Sbjct: 505 NWSTYASYTSIFQPQNDRDSSGKYLAPITGNNYELGLKSDWMNSRLTTTLAIFRIEQDNV 564 Query: 552 ATAAGR DQSGNTYYRAANQAKTHGWEIEVGGRITPEWQIQAGYSQSKPRDQDGSRLN 608 A + G +G T Y+A + + G E E+ G IT WQ+ G ++ D +G+ +N Sbjct: 565 AQSTGTPIPGSNGETAYKAVDGTVSKGVEFELNGAITDNWQLTFGATRYIAEDNEGNAVN 624 Query: 609 PDSVPERSFKLFTAYHLAPEAPSGRTIGAGVRRQGETHTDPAALRIPNPAAKARAVANSR 668 P ++P + K+FT+Y L P P T+G GV Q +TD P RA Sbjct: 625 P-NLPRTTVKMFTSYRL-PVMPE-LTVGGGVNWQNRVYTDTV TPYGTFRA E 672 Query: 669 QKAYAVADIMARYRFNPRTELSLNVDNLFNKHYRTQPDRH-SYGALRTVNAAFTYRF 724 Q +YA+ D+ RY+ L NV+NLF+K Y T + YG R + TY+F Sbjct: 673 QGSYALVDLFTRYQVTKNFSLQGNVNNLFDKTYDTNVEGSIVYGTPRNFSITGTYQF 729 Based on this analysis, it was predicted that these proteins from N. meningitidis and N.gonorrhoeae, and their epitopes, could be useful antigens for vaccines or diagnostics, or for raising antibodies. ORF23-1 (77.5kDa) was cloned in pET and pGex vectors and expressed in E.coli, as described above. The products of protein expression and purification were analyzed by SDS-PAGE. Figure 15A shows the results of affinity purification of the His-fusion protein, and Figure 15B shows the results of expression of the GST-fusion in E.coli. Purified His-fusion protein was used to immunise mice, whose sera were used for Western blot (Figure 15C) and for ELISA (positive result). These experiments confirm that ORF23-1 is a surface-exposed protein, and that it is a useful immunogen. Example 80 The following partial DNA sequence was identified in N meningitidis <SEQ ID 673>: 1 ATGCGCACGG CAGTGGTTTT GCTGTTGATC ATGCCGATGG CGGCTTCGTC 51 GGCAATGATG CCGGAAATGG TGTGCGCGGG CGTGTCGCCG GGAACGGCAA 101 TCATATCCAA GCCGACCGAA CAAACGGCGG TCATGGCTTC GAGTTTGTCC 151 AGCGTCAgcA CGCCTGCTTC GGCGgcGgCa ATCATACCTT CGTCTTCGGA 201 AACGGGGATA AACGcGCCAC TCAAACCCCC GACCGCGCTG GAAGCCATCA 251 TGCCGCCTTT TTTCACGGCA TCGTTCAGCA ATGCCAAAGC TGCTGTTGTG 301 CCGTGCGTAC CGCAGACGCT CAAGCCCATT TnTTCAAGAA TGCGTGCCAC 351 TnAGTCGCCG ACGGGG .. This coπesponds to the .amino acid sequence <SEQ ID 674; ORF24>: 1 MRTAWLLLI MPMAASSAMM PEMVCAGVSP GTAIISKPTE QTAVMASSLS 51 SVSTPASAAA IIPSSSETGI ΝAPLKPPTAL EAIMPPFFTA SFSΝAKAAW 101 PCVPQTLKPI XSRMRATXSP TG.. Further work revealed the complete nucleotide sequence <SEQ ID 675>: 1 ATGCGCACGG CAGTGGTTTT GCTGTTGATC ATGCCGATGG CGGCTTCGTC 51 GGCAATGATG CCGGAAATGG TGTGCGCGGG CGTGTCGCCG GGAACGGCAA 101 TCATATCCAA GCCGACCGAA CAAACGGCGG TCATGGCTTC GAGTTTGTCC 151 AGCGTCAGCA CGCCTGCTTC GGCGGCGGCA ATCATACCTT CGTCTTCGGA 201 AACGGGGATA AACGCGCCAC TCAAACCCCC GACCGCGCTG GAAGCCATCA 251 TGCCGCCTTT TTTCACGGCA TCGTTCAGCA ATGCCAAAGC TGCTGTTGTG 301 CCGTGCGTAC CGCAGACGCT CAAGCCCATT TCTTCAAGAA TGCGTGCCAC 351 TGAGTCGCCG ACGGCGGGGG TCGGCGCCAG CGACAAGTCG AGAATACCAA 401 ACGGGATATT CAGCATTTTT GAGGCTTCGC GGCCGATGAG TTCGCCCACG 451 CGGGTAATTT TGAAAGCAGT TTTCTTCACT ACTTCCGCAA CTTCGGTCAA 501 TGTCGTTGCA TCTGAATTTT CCAACGCGGC TTTTACGACA CCTGGGCCGG 551 ATACGCCGAC ATTGATAACG GCATCCGCTT CGCCCGAACC ATGAAACGCG 601 CCCGCCATAA ACGGGTTGTC TTCCACCGCG TTGCAGAACA CGACAATTTT 651 AGCGCAGCCG AAACCTTCGG GCGTGATTTC CGCCGTGCGT TTGACGGTTT 701 CGCCCGCCAG CTTGACCGCA TCCATATTGA TACCGGCACG CGTACTGCCG 751 ATATTGATGG AGCTGCACAC AATATCGGTA GTCTTCATCG CTTCGGGAAT 801 GGAGCGGATT AACACCTCAT CCGAAGGCGA CATCCCTTTT TGCACCAACG 851 CGGAAAAACC GCCGATAAAA GACACACCGA TGGCTTTGGC AGCTTTATCC 901 AAAGTTTGCG CCACGCTGAC GTAA This coπesponds to the .amino acid sequence <SEQ LD 676; ORF24-1 > • 1 MRTAWLLLI MPMAASSAMM PEMVCAGVSP GTAIISKPTE QTAVMASSLS 51 SVSTPASAAA IIPSSSETGI NAPLKPPTAL EAIMPPFFTA SFSNAKAAW 101 PCVPQTLKPI SSRMRATESP TAGVGASDKS RIPNGIFSIF EASRPMΞSPT 151 RVILKAVFFT TSATSVNWA SEFSNAAFTT PGPDTPTLIT ASASPEP*NA 201 PAINGLSSTA LQNTTILAQP KPSGVISAVR LTVSPASLTA SILIPARVLP 251 ILMELHTISV VFIASGMERI NTSSEGDIPF CTNAEKPPIK DTPMALAALS 301 KVCATLT* Computer analysis of this amino acid sequence gave the following results: Homology with a predicted ORF from N.meninsitidis (sfrain A) ORF24 shows 96.4% identity over a 307 aa overlap with an ORF (ORF24a) from strain A of N. meningitidis: 10 20 30 40 50 60 orf24a . pep MRTAWLLLIMPMAASSAMMPEMVCAGVSPGTAIISXPTEQTAVIASSLSNVSTPASAAA I II I I I I I I II II II I II II II I II I I I I I II I I II I I I II II : II II I : I II I I I I II orf24 MRTAWLLLIMPMAASSAMMPEMVCAGVSPGTAIISKPTEQTAVMASSLSSVSTPASAAA 10 20 30 40 50 60 70 80 90 100 110 120 orf24a . pep IIPSSSXTGINAPLKPPTALEAIMPPFFTASFSNAKAAWPCVPQTLKPISSRMRATESP I II I I I II II II I I I I I I I I I I I II I I I I I II I I II I I I I I I I I I I I I I II I I I I I I I I orf24 IIPSSSETGINAPLKPPTALEAIMPPFFTASFSNAKAAWPCVPQTLKPISSRMRATESP 70 80 90 100 110 120 130 140 150 160 170 180 orf24a. pep TAGVGASDKSRIPNGIFSIFEASRPMSSPTRVILKAVFFTTSATSVNWASEFSNAAFTT II I I I I I I II I I I II I I I I I I II I I I I I I I I II I I I I I I I I II II I II I II I I I II II I I orf24 TAGVGASDKSRIPNGIFSIFEASRPMSSPTRVILKAVFFTTSATSVNWASEFSNAAFTT 130 140 150 160 170 180 190 200 210 220 230 240 orf24a. pep PGPDTPTLITASASPEPXNAPAIXGLSSXALQNTTILAQPKPSSVISXVRLMVSPASLTA I I II I II I I I II II I II I II I I I I I I I : I I I II I I I I I I I II : II I III I I I II II I orf24 PGPDTPTLITASASPEPXNAPAINGLSSTALQNTTILAQPKPSGVISAVRLTVSPASLTA 190 200 210 220 230 240 250 260 270 280 290 300 orf24a . pep SILIPARVLPILMELHTISWFIASGMERXNTSSEGDIPFCTSAEKPPIKDTPMALAALS I II II II II II II I I II II I II I I I I II I I II I I II I I I I I : II I II I I II I I II II I I orf24 SILIPARVLPILMELHTISWFIASGMERINTSSEGDIPFCTNAEKPPIKDTPMALAALS 250 260 270 280 290 300 orf24a.pep KVCATLTX II I II I II orf24 KVCATLTX The complete length ORF24a nucleotide sequence <SEQ ID 677> is: 1 ATGCGCACGG CAGTGGTTTT GCTGTTGATC ATGCCGATGG CGGCTTCGTC 51 GGCAATGATG CCGGAAATGG TGTGCGCGGG TGTGTCGCCG GGAACGGCAA 101 TCATATCCAA NCCGACCGAA CAAACGGCGG TCATCGCTTC GAGTTTATCC 151 AACGTCAGCA CGCCTGCTTC GGCGGCGGCA ATCATACCTT CGTCTTCGGA 201 NACGGGGATA AACGCGCCAC TCAAACCGCC AACCGCGCTC GAAGCCATCA 251 TGCCGCCCTT TTTCACGGCA TCGTTCAGCA ATGCCAAAGC TGCTGTTGTG 301 CCGTGCGTAC CGCAGACGCT CAAACCCATT TCTTCAAGAA TGCGCGCCAC 351 CGAGTCGCCG ACGGCAGGGG TCGGTGCCAG CGACAAGTCG AGAATACCAA 401 ACGGGATATT CAGCATTTTT GAGGCTTCGC GGCCGATGAG TTCGCCCACG 451 CGGGTAATTT TGAAGGCGGT TTTCTTCACA ACTTCGGCAA CTTCGGTCAA 501 TGTCGTTGCA TCCGAATTTT CCAACGCGGC TTTTACGACA CCCGGGCCGG 551 ATACGCCGAC ATTAATCACA GCATCCGCTT CGCCTGAGCC GTGAAACGCG 601 CCCGCCATAN ACGGGTTGTC TTCCNCCGCG TTGCAGAACA CGACGATTTT 651 GGCGCAGCCG AAACCTTCTA GTGTGATTTC ANCCGTGCGT TTGATGGTTT 701 CGCCCGCCAG TCTGACCGCG TCCATATTGA TACCGGCGCG CGTACTGCCG 751 ATATTGATGG AGCTGCACAC GATATCAGTA GTCTTCATCG CTTCGGGAAT 801 GGAACGGATN AACACCTCGT CAGAAGGCGA CATACCTTTT TGCACCAGCG 851 CGGAAAAGCC GCCAATAAAA GACACGCCGA TGGCTTTGGC AGCCTTATCC 901 AAAGTTTGCG CCACGCTGAC GTAA This encodes a protein having amino acid sequence <SEQ ID 678>: 1 MRTAWLLLI MPMAASSAMM PEMVCAGVSP GTAIISXPTE QTAVIASSLS 51 NVSTPASAAA IIPSSSXTGI NAPLKPPTAL EAIMPPFFTA SFSNAKAAW 101 PCVPQTLKPI ΞSRMRATESP TAGVGASDKS RIPNGIFSIF EASRPMSSPT 151 RVILKAVFFT TSATSVNWA SEFSNAAFTT PGPDTPTLIT ASASPEP^NA 201 PAIXGLSSXA LQNTTILAQP KPSSVISXVR LMVSPASLTA SILIPARVLP 251 ILMELHTISV VFIASGMERX NTSSEGDIPF CTSAEKPPIK DTPMALAALS 301 KVCATLT* It should be noted that this protein includes a stop codon at position 198. ORF24a and ORF24-1 show 96.4% identity in 307 aa overlap: 10 20 30 40 50 60 orf24a. pep MRTAWLLLIMPMAASSAMMPEMVCAGVSPGTAIISXPTEQTAVIASSLSNVSTPASAAA I II I I II I II II I II I II I II I II I I II I II II I I I I I I I I I I: II I I I : II I II I I I I orf24-1 MRTAWLLLIMPMAASSAMMPEMVCAGVSPGTAIISKPTEQTAVMASSLSSVSTPASAAA 10 20 30 40 50 60 70 80 90 100 110 120 orf24a. pep IIPSSSXTGINAPLKPPTALEAIMPPFFTASFSNAKAAWPCVPQTLKPISSRMRATESP I I I I I I I I II I I I I I I II I I I I I II I I I I I I I II I II I II I I I I I I I II I I I I I II I I I orf24-1 IIPSΞSETGINAPLKPPTALEAIMPPFFTASFSNAKAAWPCVPQTLKPISSRMRATESP 70 80 90 100 110 120 130 140 150 160 170 180 orf24a . pep TAGVGASDKSRIPNGIFSIFEASRPMSSPTRVILKAVFFTTSATSVNWASEFSNAAFTT I I I I I II I I II I II I II I I I I II I I I I I I I II I I I II I I I II I II I I II II I I I I I I II I orf24-1 TAGVGASDKSRIPNGIFSIFEASRPMSSPTRVILKAVFFTTSATSVNWASEFSNAAFTT 130 140 150 160 170 180 190 200 210 220 230 240 orf24a. pep PGPDTPTLITASASPEPXNAPAIXGLSSXALQNTTILAQPKPSSVISXVRLMVSPASLTA I I I I I II I I I I I II I II I II I II I I II : II II I II I II II II : II I III II II II I I orf24-1 PGPDTPTLITASASPEPXNAPAINGLSSTALQNTTILAQPKPSGVISAVRLTVSPASLTA 190 200 210 220 230 240 250 260 270 280 290 300 orf24a. pep SILIPARVLPILMELHTISWFIASGMERXNTSΞEGDIPFCTSAEKPPIKDTPMALAALS I II I I M I II I I II I II II I I II M M I I I II I II II I II I : I II I I II II I I I II II I orf24-1 SILIPARVLPILMELHTISWFIASGMERINTSSEGDIPFCTNAEKPPIKDTPMALAALS 250 260 270 280 290 300 orf24a.pep KVCATLTX I I I I II II orf24-1 KVCATLTX Homology with a predicted ORF from N.gonorrhoeae ORF24 shows 96.7% identity over a 121 aa overlap with a predicted ORF (ORF24ng) from N.gonorrhoeae: orf24.pep MRTAWLLLIMPMAASSAMMPEMVCAGVSPGTAIISKPTEQTAVMASSLSSVSTPASAAA 60 II I II II II I II II II II II I I II I II II I II II : I I I I I I I I I I I I I I I II : II I I I I I orf24ng MRTAVVLLLIMPMAASSAMMPEMVCAGVSPGTAIMSKPTEQTAVMASSLSSVNTPASAAA 60 orf24.pep IIPSSSETGINAPLKPPTALEAIMPPFFTASFSNAKAAWPCVPQTLKPIXSRMRATXSP 120 I I I I I I II I I I II II II I I I II II II I II I I I II I I I I I I I I I I I I I I I I I I II II II orf24ng IIPSSSETGINAPLKPPTALEAIMPPFFTASFSNAKAAWPCVPQTLKPISSRMRATESP 120 orf24. pep TG 122 I : orf24ng TAGVGASDKSRMPNGIFSIFEASRPMΞSPTRVILKAVFFTTSATSVRLTASEFSSAALTT 180 The complete length ORF24ng nucleotide sequence <SEQ ID 679> is: 1 ATGCGCACGG CGGTGGTTTT GCTGTTGATC ATGCCGATGG CGGCTTCGTC 51 GGCGATGATG CCGGAAATGG TGTGCGCGGG CGTGTCGCCG GGAACGGCAA 101 TCATGTCCAA ACCAACGGAG CAGACGGCGG TCATGGCTTC GAGTTTGTCC 151 AGCGTCAACA CGCCTGCCTC GGCGGCGGCA ATCATACCTT CGTCTTCGGA 201 AACGGGGATA AACGCGCCGC TCAAACCGCC GACCGCGCTG GAAGCCATCA 251 TGCCGCCCTT TTTCACGGCA TCGTTCAGCA ATGCCAAAGC TGCTGTTGTG 301 CCGTGCGTAC CGCAGACGCT CAAGCCCATT TCTTCAAGAA TGCGCGCCAC 351 CGAGTCGCCG ACGGCGGGGG TCGGTGCCAG CGACAAATCG AGAATGCCGA 401 ACGGGATATT CAGCATTTTT GAGGCTTCGC GACCGATGAG TTCGCCCACG 451 CGGGTGATTT TGAAAGCGGT TTTCTTCACG ACTTCGGCGA CCTCGGTCAG 501 GCTGACCGCG TCCGAATTTT CCAGCGCGGC TTTGACCACG CCTGGACCGG 551 ATACGCCGAC ATTAATCACA GCATCCGCTT CGCCCGAGCC GTGGAACGCA 601 CCCGCCATAA ACGGATTGTC TTCCACCGCG TTGCAGAACA CGACGATTTT 651 GGCGCAGCCG AAACCTTCGG GTGTGATTTC AGCCGTGCGT TTGATGGTTT 701 CGCCTGCCAG CTTGACCGCA TCCATATTGA TACCGGCACG CGTGCTGCCG 751 ATATTGATGG AGCTGCACAC GATATCGGTA GTTTTCATCG CTTCGGGAAC 801 GGAACGGATC AACACCTCAT CCGAAGGCGA CATACCTTTT TGCACCAGCG 851 CGGAAAAGCC GCCGATAAAG GACACGCCGA TGGCTTTGGC TGCCTTGTCC 901 AAAGTCTGCG CCACGCTGAC ATAA This encodes a protein having amino acid sequence <SEQ ED 680>: 1 MRTAWLLLI MPMAASSAMM PEMVCAGVSP GTAIMSKPTE QTAVMASSLS 51 SVNTPASAAA IIPSSSETGI NAPLKPPTAL EAIMPPFFTA SFSNAKAAW 101 PCVPQTLKPI SSRMRATESP TAGVGASDKS RMPNGIFSIF EASRPMSSPT 151 RVILKAVFFT TSATSVRLTA SEFSSAALTT PGPDTPTLIT ASASPEPWNA 201 PAINGLSSTA LQNTTILAQP KPSGVISAVR LMVSPASLTA SILIPARVLP 251 ILMELHTISV VFIASGTERI NTSSEGDIPF CTSAEKPPIK DTPMALAALS 301 KVCATLT* ORF24ng and ORF24-1 show 96.1% identity in 307 aa overlap: 10 20 30 40 50 60 orf24-1. pep MRTAWLLLIMPMAASSAMMPEMVCAGVSPGTAIISKPTEQTAVMASSLSSVSTPASAAA I I I I I I I I I I I I I I I I I I II I I II I I II I I I I I I : I II I I I I I II II I I II I: II II I I I orf24ng MRTAWLLLIMPMAASSAMMPEMVCAGVSPGTAIMSKPTEQTAVMASSLSSVNTPASAAA 10 20 30 40 50 60 70 80 90 100 110 120 orf24-1. pep IIPSSSETGINAPLKPPTALEAIMPPFFTASFSNAKAAWPCVPQTLKPISSRMRATESP I I I I I I I II II I I I I I I I II I I I II I II I I II I I II I I II I I I II I II I I I I I I I II I I I orf24ng IIPSSSETGINAPLKPPTALEAIMPPFFTASFSNAKAAWPCVPQTLKPISSRMRATESP 70 80 90 100 110 120 130 140 150 160 170 180 orf24-1. pep TAGVGASDKSRIPNGIFSIFEASRPMSSPTRVILKAVFFTTSATSVNWASEFSNAAFTT II II I I II II I: I I I I I I II I II II II I I I I II I II I II I II II II :: II I I I: I I: II orf24ng TAGVGASDKSRMPNGIFSIFEASRPMSSPTRVILKAVFFTTSATSVRLTASEFSSAALTT 130 140 150 160 170 180 190 200 210 220 230 240 orf24-1. pep PGPDTPTLITASASPEPXNAPAINGLSSTALQNTTILAQPKPSGVISAVRLTVSPASLTA I I I II I I I I I I I I I I II I I II II I I I II II II II II I II II II I I I I I II I I I I I I I I orf24ng PGPDTPTLITASASPEPWNAPAINGLSSTALQNTTILAQPKPSGVISAVRLMVSPASLTA 190 200 210 220 230 240 250 260 270 280 290 300 orf24-1. pep SILIPARVLPILMELHTISWFIASGMERINTSSEGDIPFCTNAEKPPIKDTPMALAALS I II I II I I I I I II I II II II I II I II I II II I I I I I I I II I : I I II I I II I I I I I I II I orf24ng SILIPARVLPILMELHTISWFIASGTERINTSSEGDIPFCTSAEKPPIKDTPMALAALS 250 260 270 280 290 300 orf24-l.pep KVCATLTX I I I II I I I orf24ng KVCATLTX Based on this analysis, including the presence of a putative leader sequence (first 18 aa - double- underlined) and putative transmembrane domains (single-underlined) in the gonococcal protein, it is predicted that the proteins from N. meningitidis .and N.gonorrhoeae, .and their epitopes, could be useful antigens for vaccines or diagnostics, or for raising antibodies. Example 81 The following partial DNA sequence was identified in N. meningitidis <SEQ ID 681>: 1 ..ACCGACGTGC AAAAAGAGTT GGTCGGCGAA CAACGCAAGT GGGCGCAGGA 51 AAAAATCAGC AACTGCCGAC AAGCCGCCGC GCAGGCAGAC CGGCAGGAAT 101 ACGCCGAATA CCTCAAGCTG CAATGCGACA CGCGGATGAC GCGCGAACGG 151 ATACAGTATC TTCGCGGCTA TTCCATCGAT TAG This coπesponds to the amino acid sequence <SEQ ID 682; ORF25>: 1 ..TDVQKELVGE QRKWAQEKIS NCRQAAAQAD RQEYAEYLKL QCDTRMTRER 51 IQYLRGYSID * Further work revealed the complete nucleotide sequence <SEQ ID 683>: 1 ATGTATCGGA AACTCATTGC GCTGCCGTTT GCCCTGCTGC TTGCCGCTTG 51 CGGCAGGGAA GAACCGCCCA AGGCATTGGA ATGCGCCAAC CCCGCCGTGT 101 TGCAAGGCAT ACGCGGCAAT ATTCAGGAAA CGCTCACGCA GGAAGCGCGT 151 TCTTTCGCGC GCGAAGACGG CAGGCAGTTT GTCGATGCCG ACAAAATTAT 201 CGCCGCCGCC TACGGTTTGG CGTTTTCTTT GGAACACGCT TCGGAAACGC 251 AGGAAGGCGG GCGCACGTTC TGTATCGCCG ATTTGAACAT TACCGTGCCG 301 TCTGAAACGC TTGCCGATGC CAAGGCAAAC AGCCCCCTGT TGTACGGGGA 351 AACTGCTTTG TCGGATATTG TGCGGCAGAA GACGGGCGGC AATGTCGAGT 401 TTAAAGACGG CGTATTGACG GCAGCCGTCC GCTTCCTGCC CGTCAAAGAC 451 GGTCAGACGG CATTTGTCGA CAACACGGTC GGTATGGCGG CGCAAACGCT 501 GTCTGCCGCG CTGCTGCCTT ACGGCGTGAA GAGCATCGTG ATGATAGACG 551 GCAAGGCGGT GAAAAAAGAA GACGCGGTCA GGATTTTGAG CGGAAAAGCC 601 CGTGAAGAAG AACCGTCCAA ACCCACGCCC GAAGACATTT TGGAACACAA 651 TGCCGCCGGC GGCGATGCGG GCGTACCCCA AGCCGCAGAA GGCGCGCCCG 701 AACCGGAAAT CCTGCATCCT GACGACGGCG AGCGTGCCGA TACCGTTACC 751 GTATCACGGG GCGAAGTGGA AGAGGCGCGC GTACAAAACC AGCGTGCGGA 801 ATCCGAAATT ACCAAACTTT GGGGAGGACT CGATACCGAC GTGCAAAAAG 851 AGTTGGTCGG CGAACAACGC AAGTGGGCGC AGGAAAAAAT CAGCAACTGC 901 CGACAAGCCG CCGCGCAGGC AGACCGGCAG GAATACGCCG AATACCTCAA 951 GCTGCAATGC GACACGCGGA TGACGCGCGA ACGGATACAG TATCTTCGCG 1001 GCTATTCCAT CGATTAG This coπesponds to the amino acid sequence <SEQ LD 684; ORF25-l>: 1 MYRKLIALPF ALLLAACGRE EPPKALECAN PAVLQGIRGN IQETLTQEAR 51 SFAREDGRQF VDADKIIAAA YGLAFSLEHA SETQEGGRTF CIADLNITVP 101 SETLADAKAN SPLLYGETAL SDIVRQKTGG NVEFKDGVLT AAVRFLPVKD 151 GQTAFVDNTV GMAAQTLSAA LLPYGVKSIV MIDGKAVKKE DAVRILSGKA 201 REEEPSKPTP EDILEHNAAG GDAGVPQAAE GAPEPEILHP DDGERADTVT 251 VSRGEVEEAR VQNQRAESEI TKLWGGLDTD VQKELVGEQR KWAQEKISNC 301 RQAAAQADRQ EYAEYLKLQC DTRMTRERIQ YLRGYSID* Computer .analysis of this amino acid sequence gave the following results: Homology with a predicted ORF from N.meninsitidis (strain Al ORF25 shows 98.3% identity over a 60aa overlap with an ORF (ORF25a) from strain A of N. meningitidis: 10 20 30 orf25.pep TDVQKELVGEQRKWAQEKISNCRQAAAQAD II I I I I I I I I I I II II I I I II II I I I I II orf25a VTVSRGEVEEARVQNQRAESEITKLWGGLDTDVQKELVGEXRKWAQEKISNCRQAAAQAD 250 260 270 280 290 300 40 50 60 orf25.pep RQEYAEYLKLQCDTRMTRERIQYLRGYSIDX I I I I I I I II I I I I I II I II I I I I I I I I II I I orf25a RQEYAEYLKLQCDTRMTRERIQYLRGYSIDX 310 320 330 The complete length ORF25a nucleotide sequence <SEQ ID 685> is: 1 ATGTATCGGA AACTCATTGC GCTGCCGTTT GCCCTGCTGC TTGCCGCTTG 51 CGGCAGGGAA GAACCGCCCA AGGCATTGGA ATGCGCCAAC CCCGCCGTGT 101 TGCAANGCAT ACGCNGCAAT ATTCAGGAAA CGCTCACGCA GGAAGCGCGT 151 TCTTTCGCGC GCGAAGACNG CANGCAGTTT GTCGATGCCG ACNAAATTAT 201 CGCCGCCGCC TANGNTNNGN NGNTNTCTTT GGAACACGCT TCGGAAACGC 251 AGGAAGGCGG GCGCACGTTC TGTNTCGCCG ATTTGAACAT TACCGTGCCG 301 TCTGAAACGC TTGCCGATGC CAAGGCAAAC AGCCCCCTGC TGTACGGGGA 351 AACCGCTTTG TCGGATATTG TGCGGCAGAA GACGGGCGGC AATGTCGAGT 401 TTAAAGACGG CGTATTGACG GCAGCCGTCC GCTTCCTACC CGTCAAAGAC 451 GGTCAGANGG CATTTGTCGA CAACACGGTC GGTATGGCGG CGCAAACGCT 501 GTCTGCCGCG TTGCTGCCTT ACGGCGTGAA GAGCATCGTG ATGATAGACG 551 GCAAGGCGGT AAAAAAAGAA GACGCGGTCA GGATTNTGAG CNGANAAGCC 601 CGTGAA AAG AACCGTCCAA ANCCNNGCCC GAAGACATTT TGGAACATAA 651 TGCCGCCGGA GGGGATGCAG ACGTACCCCA AGCCGGAGAA GACGCGCCCG 701 AACCGGAAAT CCTGCATCCT GACGACGGCG AGCGTGCCGA TACCGTTACC 751 GTATCACGGG GCGAAGTGGA AGAGGCGCGN GTACAAAACC AGCGTGCGGA 801 ATCCGAAATT ACCAAACTTT GGGGAGGACT CGATACCGAC GTGCAAAAAG 851 AGTTGGTCGG CGAANAACGC AAGTGGGCGC AGGAAAAAAT CAGCAACTGC 901 CGACAAGCCG CCGCGCAGGC AGACCGGCAG GAATACGCCG AATACCTCAA 951 GCTGCAATGC GACACGCGGA TGACGCGCGA ACGGATACAG TATCTTCGCG 1001 GCTATTCCAT CGATTAG This encodes a protein having amino acid sequence <SEQ LD 686>: MYRKLIALPF ALLLAACGRE EPPKALECAN PAVLQXIRXN IQETLTQEAR 51 SFAREDXXQF VDADXIIAAA XXXXXSLEHA SETQEGGRTF CXADLNITVP 101 SETLADAKAN SPLLYGETAL SDIVRQKTGG NVEFKDGVLT AAVRFLPVKD 151 GQXAFVDNTV GMAAQTLSAA LLPYGVKSIV MIDGKAVKKE DAVRIXSXXA 201 REXEPSKXXP EDILEHNAAG GDADVPQAGE DAPEPEILHP DDGERADTVT 251 VSRGEVEEAR VQNQRAESEI TKLWGGLDTD VQKELVGEXR KWAQEKISNC 301 RQAAAQADRQ EYAEYLKLQC DTRMTRERIQ YLRGYSID* ORF25a and ORF25-1 show 93.5% identity in 338 aa overlap: 10 20 30 40 50 60 orf25a . pep MYRKLIALPFALLLAACGREEPPKALECANPAVLQXIRXNIQETLTQEARSFAREDXXQF I I I I II I II I I I I I I II I I I II I M II I II I I I II II I I I I I I I I I I I M I I I I II orf25-1 MYRKLIALPFALLLAACGREEPPKALECANPAVLQGIRGNIQETLTQEARSFAREDGRQF 10 20 30 40 50 60 70 80 90 100 110 120 orf25a. pep VDADXIIAAAXXXXXSLEHASETQEGGRTFCXADLNITVPSETLADAKANSPLLYGETAL I II I I II I I II I I I II I II II II II II II II I I I I II I I I I II II I II I I II I orf25-1 VDADKIIAAAYGLAFSLEHASETQEGGRTFCIADLNITVPSETLADAKANSPLLYGETAL 70 80 90 100 110 120 130 140 150 160 170 180 orf25a . pep SDIVRQKTGGNVEFKDGVLTAAVRFLPVKDGQXAFVDNTVGMAAQTLSAALLPYGVKSIV I II II I II II II I II II II I II II I II I I II I : I II II I I II II I I I I I I II I I I I I II I orf25-1 SDIVRQKTGGNVEFKDGVLTAAVRFLPVKDGQTAFVDNTVGMAAQTLSAALLPYGVKSIV 130 140 150 160 170 180 190 200 210 220 230 240 orf25a. pep MIDGKAVKKEDAVRIXSXXAREXEPSKXXPEDILEHNAAGGDADVPQAGEDAPEPEILHP I I I II II I I I I I I II I III I I II : II I II I I II I I I I I MMM II I I II I I I orf25-1 MIDGKAVKKEDAVRILSGKAREEEPSKPTPEDILEHNAAGGDAGVPQAAEGAPEPEILHP 190 200 210 220 230 240 250 260 270 280 290 300 orf25a . pep DDGERADTVTVSRGEVEEARVQNQRAESEITKLWGGLDTDVQKELVGEXRKWAQEKISNC I II I I I II I I I II I I I II I I I I II I I I II II II I II II I I I I I I II II I I I II I I II I I orf25-1 DDGERADTVTVSRGEVEEARVQNQRAESEITKLWGGLDTDVQKELVGEQRKWAQEKISNC 250 260 270 280 290 300 310 320 330 339 orf25a. pep RQAAAQADRQEYAEYLKLQCDTRMTRERIQYLRGYSIDX I I I II I I I I I II I I II II II I II I I I I I I I I II I II I I I orf25-l RQAAAQADRQEYAEYLKLQCDTRMTRERIQYLRGYSIDX 310 320 330 Homology with a predicted ORF from N.gonorrhoeae ORF25 shows 100% identity over a 60aa overlap with a predicted ORF (ORF25ng) from N.gonorrhoeae: orf25. pep TDVQKELVGEQRKWAQEKISNCRQAAAQAD 30 I I I II I II I I I I I I I I II II I I I II II II I orf25ng VTVSRGEVEEARVQNQRAESEITKLWGGLDTDVQKELVGEQRKWAQEKISNCRQAAAQAD 308 orf25.pep RQEYAEYLKLQCDTRMTRERIQYLRGYSID 60 II II I II I II II II I I I I I II I II I II I II orf25ng RQEYAEYLKLQCDTRMTRERIQYLRGYSID 338 The complete length ORF25ng nucleotide sequence <SEQ ED 687> is: 1 ATGTATCGGA AACTCATTGC GCTGCCGTTT GCCCTGCTGC TTGCAGCGTG 51 CGGCAGGGAA GAACCGCCCA AGGCGTTGGA ATGCGCCAAC CCCGCCGTGT 101 TGCAGGACAT ACGCGGCAGT ATTCAGGAAA CGCTCACGCA GGAAGCGCGT 151 TCTTTCGCGC GCGAAGACGG CAGGCAGTTT GTCGATGCCG ACAAAATTAT 201 CGCCGCCGCC TACGGTTTGG CGTTTTCTTT GGAACACGCT TCGGAAACGC 251 AGGAAGGCGG GCGCACGTTC TGTATCGCCG ATTTGAACAT TACCGTGCCG 301 TCTGAAACGC TTGCCGATGC CGAGGCAAAC AGCCCCCTGC TGTATGGGGA 351 AACGTCTTTG GCAGACATCG TGCAGCAGAA GACGGGCGGC AATGTCGAGT 401 TTAAAGACGG CGTATTGACG GCAGCCGTCC GCTTCCTGCC CGCCAAAGAC 451 GCTCGGACGG CATTTATCGA CAACACGGTC GGTATGGCGA CGCAAACGCT 501 GTCTGCCGCG TTGCTGCCTT ACGGCGTGAA GAGCATCGTG ATGATAGACG 551 GCAAGGCGGT GACAAAAGAA GACGCGGTCA GGGTTTTGAG CGGCAAAGCC 601 CGTGAAGAAG AACCGTCCAA ACCCACCCCC GAAGACATTT TGGAACACAA 651 TGCCGCCGGC GGCGATGCGG GCGTACCCCA AGCCGCAGAA GGCGCACCCG 701 AACCCGAAAT CCTGCATCCC GACGACGTCG AGCGTGCCGA TACCGTTACC 751 GTATCACGGG GCGAAGTGGA AGAGGCGCGC GTACAAAACC AACGTGCGGA 801 ATCCGAAATT ACCAAACTTT GGGGAGGACT CGATACCGAC GTGCAAAAAG 851 AGTTGGTCGG CGAACAGCGC AAGTGGGCGC AGGAAAAAAT CAGcaactgc 901 cgACAAGCCG CCGCGCAGGC AGACCGGCAG GAATACGCCG AATACCTCAA 951 GCTCCAATGC GACACGCGGA TGACGCGCGA ACggaTACAG TATCTTCGCG 1001 GCTATTCCAT CGATTAG This encodes a protein having amino acid sequence <SEQ LD 688>: 1 MYRKLIALPF ALLLAACGRE EPPKALECAN PAVLQDIRGS IQETLTQEAR 51 SFAREDGRQF VDADKIIAAA YGLAFSLEHA SETQEGGRTF CIADLNITVP 101 SETLADAEAN SPLLYGETSL ADIVQQKTGG NVEFKDGVLT AAVRFLPAKD 151 ARTAFIDNTV GMATQTLSAA LLPYGVKSIV MIDGKAVTKE DAVRVLSGKA 201 REEEPSKPTP EDILEHNAAG GDAGVPQAAE GAPEPEILHP DDVERADTVT 251 VSRGEVEEAR VQNQRAESEI TKLWGGLDTD VQKELVGEQR KWAQEKISNC 301 RQAAAQADRQ EYAEYLKLQC DTRMTRERIQ YLRGYSID* ORF25ng and ORF25-1 show 95.9% identity in 338 aa overlap: 10 20 30 40 50 60 orf25-1. pep MYRKLIALPFALLLAACGREEPPKALECANPAVLQGIRGNIQETLTQEARSFAREDGRQF I I I II I I I I I I I I I I II I II I II I I I II I I II I I I I I I : I I II I I I I I I I II I I I II I I orf25ng MYRKLIALPFALLLAACGREEPPKALECANPAVLQDIRGSIQETLTQEARSFAREDGRQF 10 20 30 40 50 60 70 80 90 100 110 120 orf25-1. pep VDADKIIAAAYGLAFSLEHASETQEGGRTFCIADLNITVPSETLADAKANSPLLYGETAL I I I II I I I II II II I I I II I I I I II I I I II II I I I I II I II I II I I I : I I I I II I I II : I orf25ng VDADKIIAAAYGLAFSLEHASETQEGGRTFCIADLNITVPSETLADAEANSPLLYGETSL 70 80 90 100 110 120 130 140 150 160 170 180 orf25-1. pep SDIVRQKTGGNVEFKDGVLTAAVRFLPVKDGQTAFVDNTVGMAAQTLSAALLPYGVKSIV : II I : I I I I I I I II I I I II I I I I II I I : II :: I II : I I II I II : II I II I I I I I I I I I II orf25ng ADIVQQKTGGNVEFKDGVLTAAVRFLPAKDARTAFIDNTVGMATQTLSAALLPYGVKSIV 130 140 150 160 170 180 190 200 210 220 230 240 orf25-1. pep MIDGKAVKKEDAVRILSGKAREEEPSKPTPEDILEHNAAGGDAGVPQAAEGAPEPEILHP II I II I I I I I II I : I II I II I II I I I I I I II I I I I I I I II II I II I I I I I I I I I I I I II orf25ng MIDGKAVTKEDAVRVLSGKAREEEPSKPTPEDILEHNAAGGDAGVPQAAEGAPEPEILHP 190 200 210 220 230 240 250 260 270 280 290 300 orf25-1. pep DDGERADTVTVSRGEVEEARVQNQRAESEITKLWGGLDTDVQKELVGEQRKWAQEKISNC II I I I II II II I II I II II I II I I II I I I II I I I I I I II I II I II I I I I II I I I I I I II orf25ng DDVERADTVTVSRGEVEEARVQNQRAESEITKLWGGLDTDVQKELVGEQRKWAQEKISNC 250 260 270 280 290 300 310 320 330 339 orf25-l.pep RQAAAQADRQEYAEYLKLQCDTRMTRERIQYLRGYSIDX II I II I I II II I I I II I II I I I I I II I I I II II I II II I orf25ng RQAAAQADRQEYAEYLKLQCDTRMTRERIQYLRGYSIDX 310 320 330 Based on this analysis, including the presence of a predicted prokaryotic membrane lipoprotein lipid attchment site (underlined) in the gonococcal protem, it was predicted that the proteins from N. meningitidis .and N.gonorrhoeae, and their epitopes, could be useful antigens for vaccines or diagnostics, or for raising antibodies. ORF25-1 (37kDa) was cloned in pET and pGex vectors and expressed in E.coli, as described above. The products of protein expression and purification were analyzed by SDS-PAGE. Figure 16A shows the results of affinity purification of the GST-fusion protein, and Figure 16B shows the results of expression of the His-fusion in E.coli. Purified His-fusion protein was used to immunise mice, whose sera were used for Western blot (Figure 16C), ELISA (positive result), and FACS analysis (Figure 16D). These experiments confirm that ORF25-1 is a surface-exposed protein, and that it is a useful immunogen. Figure 16E shows plots of hydrophilicity, antigenic index, and AMPHI regions for ORF25-1. Example 82 The following partial DNA sequence was identified in N. meningitidis <SEQ ID 689> 1 ATGCAGCTGA TCGACTATTC ACATTCATTT TTCTCGGTTG TGCCACCCTT 51 TTTGGCACTG GCACTTGCCG TCATTACCCG CCGCGTACTG CTGTCTTTAG 101 GCATCGGTAT TCTGGwysGC GTTGCCTTTT TGGTCGGCGG CAACCCCGTC 151 GACGGTCTGA CACACCTGAA AGACATGGTC GTCGGCTTGG CTTGGTCAGA 201 CGsyGATTGG TCGCTGGGCA AACCAAAAAT CTTGGTTTTC CkGATACTTT 251 TGGGTATTTT TACTTCCCTG CTGACCTACT CCGGCAGCAA T // 851 AC TTCGCTGGTA 901 TTCGGCGGCA CTTGCGGCGT CTTTGCCGTC GTTCTCTGCA CGCTCGGCAC 951 GATTAAAACC GCCGACTATC CCAAAGCCGT TTGGCAGGGT GCGAAATCTA 1001 TGTTCGGCGC AATCGCCATT TTAATCCTCG CTTGGCTCAT CAGTACGGTT 1051 GTCGGCGAAA TGCACACCGG CGATTACCTC TCCACACTGG TTGCGGGCAA 1101 CATCCATCCC GGCTTCCTGC CCGTCATCCT CTTCCTGCTC GCCAGCGTGA 1151 TGGCGTTTGC CACAGGCACA AGCTGGGGGA CGTTCGGCAT TATGCTGCCG 1201 ATTGCCGCCG CCATGGCGGT CAAAGTCGAA CCCGCGCTGA TTATCCCGTG 1251 TATGTCCGCA GTAATGGCGG GGGCGGTATG CGGCGACCAC TGCTCGCCCA 1301 TTTCCGACAC GACCATCCTG TCGTCCACCG GCGCGCGCTG CAACCACATC 1351 GACCACGTTA CCTCGCAACT GCCTTACGCC TTAACCGTTG CCGCCGCCGC 1401 CGCATCGGGC TACCTCGCAT TGGGTCTGAC AAAATCCGCG CTGTTGGGCT 1451 TTGGCACGAC AGGCATTGTA TTGGCGGTGC TGATTTTTCT GTTGAAAGAT 1501 AAAAAA.. This coπesponds to the amino acid sequence <SEQ ID 690; ORF26>: 1 MQLIDYSHSF FSWPPFLAL ALAVITRRVL LSLGIGILXX VAFLVGGNPV 51 DGLTHLKDMV VGLAWSDXDW SLGKPKILVF XILLGIFTSL LTYSGSN ... // 251 TSLV 301 FGGTCGVFAV VLCTLGTIKT ADYPKAVWQG AKSMFGAIAI LILAWLISTV 351 VGEMHTGDYL STLVAGNIHP GFLPVILFLL ASVMAFATGT SWGTFGIMLP 401 IAAAMAVKVE PALIIPCMSA VMAGAVCGDH CSPISDTTIL SSTGARCNHI 451 DHVTSQLPYA LTVAAAAASG YLALGLTKSA LLGFGTTGIV LAVLIFLLKD 501 KK.. Further work revealed the complete nucleotide sequence <SEQ ID 691>: 1 ATGCAGCTGA TCGACTATTC ACATTCATTT TTCTCGGTTG TGCCACCCTT 51 TTTGGCACTG GCACTTGCCG TCATTACCCG CCGCGTACTG CTGTCTTTAG 101 GCATCGGTAT TCTGGTCGGC GTTGCCTTTT TGGTCGGCGG CAACCCCGTC 151 GACGGTCTGA CACACCTGAA AGACATGGTC GTCGGCTTGG CTTGGTCAGA 201 CGGCGATTGG TCGCTGGGCA AACCAAAAAT CTTGGTTTTC CTGATACTTT 251 TGGGTATTTT TACTTCCCTG CTGACCTACT CCGGCAGCAA TCAGGCGTTT 301 GCCGACTGGG CAAAACGGCA CATTAAAAAC CGGCGCGGCG CGAAAATGCT 351 GACCGCCTGC CTCGTGTTCG TAACCTTTAT CGACGACTAT TTCCACAGTC 401 TCGCCGTCGG TGCGATTGCC CGCCCCGTTA CCGACAAGTT TAAAGTTTCC 451 CGCACCAAAC TCGCCTACAT CCTCGACTCC ACTGCCGCTC CTATGTGCGT 501 GCTGATGCCC GTTTCAAGCT GGGGCGCGTC GATTATCGCC ACGCTTGCCG 551 GACTGCTCGT TACCTACAAA ATCACCGAAT ACACGCCGAT GGGGACGTTT 601 GTCGCCATGA GCCTGATGAA CTATTACGCA CTGTTTGCCC TGATTATGGT 651 GTTCGTCGTC GCATGGTTTT CCTTCGACAT CGGCTCGATG GCACGTTTCG 701 AACAAGCCGC GTTGAACGAA GCCCACGATG AAACTGCCGT TTCAGACGCT 751 ACCAAAGGTC GTGTTTACGC ACTGATTATT CCCGTTTTGG CCTTAATCGC 801 CTCAACGGTT TCCGCCATGA TCTACACCGG CGCGCAGGCA AGCGAAACCT 851 TCAGCATTTT GGGGGCATTT GAAAACACGG ACGTAAACAC TTCGCTGGTA 901 TTCGGCGGCA CTTGCGGCGT CCTTGCCGTC GTTCTCTGCA CGCTCGGCAC 951 GATTAAAACC GCCGACTATC CCAAAGCCGT TTGGCAGGGT GCGAAATCTA 1001 TGTTCGGCGC AATCGCCATT TTAATCCTCG CTTGGCTCAT CAGTACGGTT 1051 GTCGGCGAAA TGCACACCGG CGATTACCTC TCCACACTGG TTGCGGGCAA 1101 CATCCATCCC GGCTTCCTGC CCGTCATCCT CTTCCTGCTC GCCAGCGTGA 1151 TGGCGTTTGC CACAGGCACA AGCTGGGGGA CGTTCGGCAT TATGCTGCCG 1201 ATTGCCGCCG CCATGGCGGT CAAAGTCGAA CCCGCGCTGA TTATCCCGTG 1251 TATGTCCGCA GTAATGGCGG GGGCGGTATG CGGCGACCAC TGCTCGCCCA 1301 TTTCCGACAC GACCATCCTG TCGTCCACCG GCGCGCGCTG CAACCACATC 1351 GACCACGTTA CCTCGCAACT GCCTTACGCC TTAACCGTTG CCGCCGCCGC 1401 CGCATCGGGC TACCTCGCAT TGGGTCTGAC AAAATCCGCG CTGTTGGGCT 1451 TTGGCACGAC AGGCATTGTA TTGGCGGTGC TGATTTTTCT GTTGAAAGAT 1501 AAAAAACGCG CCAACGCCTG A This coπesponds to the .amino acid sequence <SEQ ID 692; ORF26-l>: 1 MQLIDYSHSF FSWPPFLAL ALAVITRRVL LSLGIGILVG VAFLVGGNPV 51 DGLTHLKDMV VGLAWSDGDW SLGKPKILVF LILLGIFTSL LTYSGSNQAF 101 ADWAKRHIKN RRGAKMLTAC LVFVTFIDDY FHSLAVGAIA RPVTDKFKVS 151 RTKLAYILDS TAAPMCVLMP VSSWGASIIA TLAGLLVTYK ITEYTPMGTF 201 VAMSLMNYYA LFALIMVFW AWFSFDIGSM ARFEQAALNE AHDETAVSDA 251 TKGRVYALII PVLALIASTV SAMIYTGAQA SETFSILGAF ENTDVNTSLV 301 FGGTCGVLAV VLCTLGTIKT ADYPKAVWQG AKSMFGAIAI LILAWLISTV 351 VGEMHTGDYL STLVAGNIHP GFLPVILFLL ASVMAFATGT SWGTFGIMLP 401 lAAAMAVKVE PALIIPCMSA VMAGAVCGDH CSPISDTTIL SSTGARCNHI 451 DHVTSQLPYA LTVAAAAASG YLALGLTKSA LLGFGTTGIV LAVLIFLLKD 501 KKRANA* Computer analysis of this amino acid sequence gave the following results: Homology with the hypothetical transmembr.ane protein HI1586 of H.influenzae (accession number P44263) ORF26 and HI1586 show 53% and 49% amino acid identity in 97 and 221 aa overlap at the N-terminus and C-terminus, respectively: Orf 26 1 MQLIDYSHSFFSWPPFLALALAVITRRVXXXXXXXXXXXVAFLVGGNPVDGLTHLKDMV 60 M+LID+S S +S+VP LA+ LA+ TRRV L +L V HI1586 14 MELIDFSSSVWSIVPALLAIILAIATRRVLVSLSAGIIIGSLMLSDWQIGSAFNYLVKNV 73 Orf 26 61 VGLAWSDXDWSLGKPKILVFXILLGIFTSLLTYSGSN 97 V L ++D + + I++F +LLG+ T+LLT SGSN HI 1586 74 VSLVYADGEIN-SNMNIVLFLLLLGVLTALLTVSGSN 109 // Orf26 86 IFTSLLTYSGS— NTSLVFGGTCGVFAWLCTL— GTIKTADYPKAVWQGAKSMFGXXXX 141 +F+ L T+ + TSLV GG C + L + + +Y ++ G KSM G HI1586 299 VFSVLGTFENTWGTSLWGGFCSIIISTLLIILDRQVSVPEYVRSWIVGIKSMSGAIAI 358 Orf26 142 XXXXXXXSTWGEMHTGDYLSTLVAGNIHPGFLPVILFLLASVMAFATGTSWGTFGIMLP 201 + +VG+M TG YLS+LV+GNI FLPVILF+L + MAF+TGTSWGTFGIMLP HI 1586 359 LFFAWTINKIVGDMQTGKYLSSLVSGNIPMQFLPVILFVLGAAMAFSTGTSWGTFGIMLP 418 Orf 26 202 IAAAMAVKVEPALIIPCMSAVMAGAVCGDHCSPISDTTILSSTGARCNHIDHVTSQXXXX 261 IAAAMA P L++PC+SAVMAGAVCGDHCSP+SDTTILSSTGA+CNHIDHVT+Q HI1586 419 IAAAMAANAAPELLLPCLSAVMAGAVCGDHCSPVSDTTILSSTGAKCNHIDHVTTQLPYA 478 0rf26 262 XXXXXXXXXXXXXXXXXKSALLGFGTTGIVLAVLIFLLKDK 302 S L GF T + L V+IF +K + HI 1586 479 ATVATATSIGYIWGFTYSGLAGFAATAVSLIVIIFAVKKR 519 Homology with a predicted ORF from Nmeningitidis (strain A) ORF26 shows 58.2% identity over a 502aa overlap with an ORF (ORF26a) from strain A of N. meningitidis: 10 20 30 40 50 60 orf26. pep MQLIDYΞHSFFSWPPFLALALAVITRRVLLSLGIGILXXVAFLVGGNPVDGLTHLKDMV I I II I II I II I I I I I I I I II I I I I I II II I II I II I II II I I I II I I I I I II II I I I I orf 26a MQLIDYSHSFFSWPPFLALALAVITRRVLLSLGIGILVGVAFLVGGNPVDGLTHLKDMV 10 20 30 40 50 60 70 80 90 99 orf 26 . pep VGLAWSDXDWSLGKPKILVFXILLGIFTSLLTYSGSNXX I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I or f 26a VGLAWSDGDWSLGKPKXLVFLILLGIFTSLLTYSGSNQAFADWAKRHIKNRRGAKMLTAC 70 80 90 100 110 120 orf26. pep orf26a LVFVTFIDDYFHSLAVGAXARPVTDKFKVSRAKLAYILDSTAAPMCVLMPVSSWGASIIA 130 140 150 160 170 180 orf26. pep orf26a TLAGLLVTYKITEYTPMGTFVAMSLMNYYALFALIMVFWAWFSFDIGSMARFEQAALNE 190 200 210 220 230 240 100 110 orf26. pep TSLV I I I I orf26a AHDETAVSDGSWGRVYALIIPVLALIASTVSAMIYTGAQASETFSILGAFENTDVNTSLV 250 260 270 280 290 300 120 130 140 150 160 170 orf26.pep FGGTCGVFAWLCTLGTIKTADYPKAVWQGAKSMFGAIAILILAWLISTWGEMHTGDYL I II I I II: I I I II I I I I II I I I I I I I I II I I II I I I I II I II I I I I I I I I I I I I II II I orf26a FGGTCGVLAWLCTLGTIKIADYPKAVWQGAKSMFGAIAILILAWLISTWGEMHTGDYL 310 320 330 340 350 360 180 190 200 210 220 230 orf26. pep STLVAGNIHPGFLPVILFLLASVMAFATGTSWGTFGIMLPIAAAMAVKVEPALIIPCMSA I II I I II II I II I II I II I I II II I II I I I I I II II II I I I I II I I I I : I : II I I I I I I orf26a STLVAGNIHPGFLXVILFLLASVMAFATGTSWGTFGIMLPIAAAMAVKVDPSLIIPCMSA 370 380 390 400 410 420 240 250 260 270 280 290 orf26. pep VMAGAVCGDHCSPISDTTILSSTGARCNHIDHVTSQLPYALTVAAAAASGYLALGLTKSA II I I I I I I I I II I I I I I II I I I I II I II I I I I I II I I I I II I I I I II I II I I I II I I I I I orf26a VMAGAVCGDHCSPISDTTILSSTGARCNHIDHVTSQLPYALTVAAAAASGYLALGLTKSA 430 440 450 460 470 480 300 310 orf26. pep LLGFGTTGIVLAVLIFLLKDKK M I I I : I M I M I I I I I II I II orf26a LLGFGXTGIVLAVLIFLLKDKKRANAX 490 500 The complete length ORF26a nucleotide sequence <SEQ LD 693> is: 1 ATGCAGCTGA TCGACTATTC ACATTCATTT TTCTCGGTTG TGCCACCCTT 51 TTTGGCACTG GCACTTGCCG TCATTACCCG CCGCGTACTG CTGTCTTTAG 101 GCATCGGTAT TCTGGTCGGC GTTGCCTTTT TGGTCGGCGG CAACCCCGTC 151 GACGGTCTGA CACACCTGAA AGACATGGTC GTCGGCTTGG CTTGGTCAGA 201 CGGCGATTGG TCGCTGGGCA AACCAAAANT CTTGGTTTTC CTGATACTTT 251 TGGGTATTTT TACTTCCCTG CTGACCTACT CCGGCAGCAA TCAGGCGTTT 301 GCCGACTGGG CAAAACGGCA CATTAAAAAC CGGCGCGGCG CGAAAATGCT 351 GACCGCCTGC CTCGTGTTCG TAACCTTTAT CGACGACTAT TTCCACAGTC 401 TCGCCGTCGG TGCGNTTGCC CGCCCCGTTA CCGACAAGTT TAAAGTTTCC 451 CGCGCCAAAC TCGCCTACAT CCTCGACTCC ACTGCCGCGC CTATGTGCGT 501 GCTGATGCCC GTTTCAAGCT GGGGCGCGTC GATTATCGCC ACGCTTGCCG 551 GACTGCTCGT TACCTACAAA ATCACCGAAT ACACGCCGAT GGGGACGTTT 601 GTCGCCATGA GCCTGATGAA CTATTACGCA CTGTTTGCCC TGATTATGGT 651 GTTCGTCGTC GCATGGTTCT CCTTCGACAT CGGCTCGATG GCACGTTTCG 701 AACAAGCCGC GTTGAACGAA GCCCACGATG AAACTGCCGT TTCAGACGGC 751 AGCTGGGGCA GGGTTTACGC ATTGATTATT CCCGTTTTGG CCTTAATCGC 801 CTCAACGGTT TCCGCCATGA TCTACACCGG TGCACAGGCA AGCGAAACCT 851 TCAGCATTTT GGGTGCATTT GAAAATACGG ACGTGAACAC TTCGCTGGTA 901 TTCGGCGGCA CTTGCGGCGT GCTTGCCGTC GTCCTCTGCA CGCTCGGCAC 951 GATTAAAATC GCCGATTATC CCAAAGCCGT TTGGCAGGGT GCGAAATCCA 1001 TGTTCGGCGC AATCGCCATT TTAATCCTTG CCTGGCTCAT CAGTACGGTT 1051 GTCGGCGAAA TGCACACAGG CGACTACCTC TCCACGCTGG TTGCGGGCAA 1101 CATCCATCCC GGCTTCCTGN CCGTCATCCT TTTCCTGCTC GCCAGCGTGA 1151 TGGCGTTTGC CACAGGCACA AGCTGGGGGA CGTTCGGCAT CATGCTGCCG 1201 ATTGCCGCCG CCATGGCGGT CAAAGTCGAT CCCTCACTGA TTATCCCGTG 1251 TATGTCCGCC GTGATGGCGG GGGCGGTATG CGGCGACCAC TGCTCGCCCA 1301 TTTCCGACAC GACCATCCTG TCGTCCACCG GCGCGCGCTG CAACCACATC 1351 GACCACGTTA CNTCGCAACT GCCTTACGCC TTAACCGTTG CCGCCGCCGC 1401 CGCATCGGGN TACCTCGCAT TGGGTCTGAC AAAATCCGCG CTGTTGGGTT 1451 TTGGCANGAC AGGCATTGTA TTGGCGGTGC TGATTTTTCT GTTGAAAGAT 1501 AAAAAACGCG CCAACGCCTG A This encodes a protein having amino acid sequence <SEQ LD 694>: 1 MQLIDYSHSF FSWPPFLAL ALAVITRRVL LSLGIGILVG VAFLVGGNPV 51 DGLTHLKDMV VGLAWSDGDW SLGKPKXLVF LILLGIFTSL LTYSGSNQAF 101 ADWAKRHIKN RRGAKMLTAC LVFVTFIDDY FHSLAVGAXA RPVTDKFKVS 151 RAKLAYILDS TAAPMCVLMP VSSWGASIIA TLAGLLVTYK ITEYTPMGTF 201 VAMSLMNYYA LFALIMVFW AWFSFDIGSM ARFEQAALNE AHDETAVSDG 251 SWGRVYALII PVLALIASTV SAMIYTGAQA SETFSILGAF ENTDVNTSLV 301 FGGTCGVLAV VLCTLGTIKI ADYPKAVWQG AKSMFGAIAI LILAWLISTV 351 VGEMHTGDYL STLVAGNIHP GFLXVILFLL ASVMAFATGT SWGTFGIMLP 401 IAAAMAVKVD PSLIIPCMSA VMAGAVCGDH CSPISDTTIL SSTGARCNHI 451 DHVTSQLPYA LTVAAAAASG YLALGLTKSA LLGFGXTGIV LAVLIFLLKD 501 KKRANA* ORF26a and ORF26-1 show 97.8% identity in 506 aa overlap: 10 20 30 40 50 60 orf26a. pep MQLIDYSHSFFS PPFLALALAVITRRVLLSLGIGILVGVAFLVGGNPVDGLTHLKDMV I II II I II II II I II I II I II I II I I I I I I II I I I I II II II II I I II I II I I I I I II I I orf26-1 MQLIDYSHSFFSVVPPFLALALAVITRRVLLSLGIGILVGVAFLVGGNPVDGLTHLKDMV 10 20 30 40 50 60 70 80 90 100 110 120 orf26a . pep VGLAWSDGDWSLGKPKXLVFLILLGIFTSLLTYSGSNQAFADWAKRHIKNRRGAKMLTAC I I II I I I II II I I I I I II I I II I II II I II I I II II II II I I II I I I I I II II I II I II orf26-1 VGLAWSDGDWSLGKPKILVFLILLGIFTSLLTYSGSNQAFADWAKRHIKNRRGAKMLTAC 70 80 90 100 110 120 130 140 150 160 170 180 orf26a. pep LVFVTFIDDYFHSLAVGAXARPVTDKFKVSRAKLAYILDSTAAPMCVLMPVSSWGASIIA I II II I II II II II II I I I II I I II II I IM I I I II II I I I I II I I II I I II I I I II I I orf26-1 LVFVTFIDDYFHSLAVGAIARPVTDKFKVSRTKLAYILDSTAAPMCVLMPVSSWGASIIA 130 140 150 160 170 180 190 200 210 220 230 240 orf26a . pep TLAGLLVTYKITEYTPMGTFVAMSLMNYYALFALIMVFWAWFSFDIGSMARFEQAALNE I II I II II II I I I I I I I I I I I II I I I I II I II I I I I II I I I I I I I I I I I II I I I I I I II I orf26-1 TLAGLLVTYKITEYTPMGTFVAMSLMNYYALFALIMVFWAWFSFDIGSMARFEQAALNE 190 200 210 220 230 240 250 260 270 280 290 300 orf26a. pep AHDETAVSDGSWGRVYALIIPVLALIASTVSAMIYTGAQASETFSILGAFENTDVNTSLV I II I II II M: I I I II I II I I II I I I I I I II II I I I I I II II I II I I II II II II I I II orf26-1 AHDETAVSDATKGRVYALIIPVLALIASTVSAMIYTGAQASETFSILGAFENTDVNTSLV 250 260 270 280 290 300 310 320 330 340 350 360 orf26a. pep FGGTCGVLAWLCTLGTIKIADYPKAVWQGAKSMFGAIAILILAWLISTWGEMHTGDYL II I II I I I II II I I I II II I II II I I I I I II I I II I I I I I I I I II I I II II I I II I II I orf26-1 FGGTCGVLAWLCTLGTIKTADYPKAVWQGAKSMFGAIAILILAWLISTWGEMHTGDYL 310 320 330 340 350 360 370 380 390 400 410 420 orf26a. pep STLVAGNIHPGFLXVILFLLASVMAFATGTSWGTFGIMLPIAAAMAVKVDPSLIIPCMSA II I I I I I I I I II I I I I II I I II I I I I I I I II I I II I II I I I I I II I I I : I : II II I I II orf26-l STLVAGNIHPGFLPVILFLLASVMAFATGTSWGTFGIMLPIAAAMAVKVEPALIIPCMSA 370 380 390 400 410 420 430 440 450 460 470 480 orf26a.pep VMAGAVCGDHCSPIΞDTTILSSTGARCNHIDHVTSQLPYALTVAAAAASGYLALGLTKSA I I II I I I II I I I I I II II I I I I I I I II I I I I II I II I I I I I I I I II I I I I I II I II II II orf26-l VMAGAVCGDHCSPISDTTILSSTGARCNHIDHVTSQLPYALTVAAAAASGYLALGLTKSA 430 440 450 460 470 480 490 500 orf26a. pep LLGFGXTGIVLAVLIFLLKDKKRANAX I I I I M M I I I I I I I I I I I I I I I I I I I orf26-l LLGFGTTGIVLAVLIFLLKDKKRANAX 490 500 Homology with a predicted ORF from Nsonorrhoeae ORF26 shows 94.8% .and 99% identity in 97 .and 206 aa overlap at the N-terminus and C-terminus, respectively, with a predicted ORF (ORF26ng) from N gonorrhoeae: orf26. pep MQLIDYSHSFFSWPPFLALALAVITRRVLLSLGIGILXXVAFLVGGΝPVDGLTHLKDMV 60 II II I II II I II II I I I II II I II II II I II I I II II I II I II I I I I II I I II II II I orf26ng MQLIDYSHSFFSWPPFLALALAVITRRVLLSLGIGILVGVAFLVGGΝPVDGLTHLKDMV 60 orf26.pep VGLAWSDXDWSLGKPKILVFXILLGIFTSLLTYSGSΝ 97 I II I I : I I I I I I I II II I I II I I I I I II I I I II II orf26ng VGLAWADGDWSLGKPKILVFLILLGIFTSLLTYSGSΝQAFADWAKRHIKΝRCGAKMLTAC 120 // orf26.pep TSLVFGGTCGVFAWLCTLGTIKTADYPKA 326 I I II II I I II M I I I I I I : I II I II II II I orf26ng ASTVSAMIYTGAQASETFSILGAFEΝTDVΝTSLVFGGTCGVLAWLCTFGTIKTADYPKA 326 orf26.pep VWQGAKSMFGAIAILILAWLISTWGEMHTGDYLSTLVAGΝIHPGFLPVILFLLASVMAF 386 II I I I II I II I I I I I I II I I I I I I II II I I I II I I I II I I I I I I I I II I I I I II I I I II I orf26ng VWQGAKSMFGAIAILILAWLISTWGEMHTGDYLSTLVAGΝIHPGFLPVILFLLASVMAF 386 orf26.pep ATGTSWGTFGIMLPIAAAMAVKVEPALIIPCMSAVMAGAVCGDHCSPISDTTILSSTGAR 446 I I II I I I II I I I II I I I II I II I II I I I II I I I II I I I I II I II II II I I I II I I I I I I I orf26ng ATGTSWGTFGIMLPIAAAMAVKVEPALIIPCMSAVMAGAVCGDHCSPISDTTILSSTGAR 446 orf 26 . pep CΝHIDHVTSQLPYALTVAAAAASGYLALGLTKSALLGFGTTGIVLAVLIFLLKDKK 502 I I orf26ng CΝHIDHVTSQLPYALTVAAAAASGYLALGLTKSALLGFGTTGIVLAVLIFLLKDKKRADV 506 The complete length ORF26ng nucleotide sequence <SEQ ID 695> is: 1 ATGCAGCTGA TTGACTATTC ACATTCATTT TTCTCGGTTG TGCCACCCTT 51 TTTGGCACTG GCACTTGCCG TCATTACCCG CCGCGTACTG CTGTCTTTAG 101 GCATCGGTAT TTTGGTCGGC GTTGCCTTTT TGGTCGGCGG CAACCCCGTC 151 GACGGTCTGA CACACCTGAA AGACATGGTC GTCGGCTTGG CTTGGGCAGA 201 CGGCGATTGG TCGCTGGGCA AACCAAAAAT CTTGGTTTTC CTGATACTTT 251 TGGGCATTTT CACTTCACTG CTGACCTACT CCGGCAGCAA TCAGGCGTTT 301 GCCGACTGGG CAAAACGGCA CATTAAAAAC CGGTGCGGCG CGAAAATGCT 351 GACCGCCTGC CTCGTGTTCG TAACCTTTAT CGACGACTAT TTCCACAGCC 401 TCGCCGTCGG TGCGATTGCC CGCCCCGTTA CCGACAAGTT TAAAGTTTCC 451 CGCGCCAAAC TCGCCTACAT CCTCGACTCC ACTGCCTCGC CCATGTGCGT 501 GCTGATGCCC GTTTCAAGCT GGGGCGCGTC GATTATCGCC ACGCTTGCCG 551 GATTGCTCGT TACCTACAAA ATTACCGAAT ACACGCCGAT GGGGACGTTT 601 GTCGCCATGA GCCTGATGAA CTATTACGCG CTGTTTGCCC TGATTATGGT 651 ATTCGTCGTC GCATGGTTCT CCTTCGACAT CGGCTCGAtg gCGCGTTTCG 701 AACAGGCTGC GTTGAACGAA gcccaggacg aaaccgccgc tTCAGACgCT 751 ACCAAAGGTC GTGTTTACGC ATTGATTATT CCCGTTTTGG CCTTAATCGC 801 CTCAACGGTT TCCGCCATGA TCTACACCGG CGCGCAGGCA AGCGAAACCT 851 TCAGCATTTT GGGGGCATTT GAAAATACCG ACGTAAACAC TTCGCTGGTA 901 TTCGGCGGCA CTTGCGGCGT GCTTGCCGTC GTCCTCTGCA CGTTCGGCAC 951 GATTAAAACC GCCGATTATC CCAAAGCCGT GTGGCAGGGT GCGAAATCCA 1001 TGTTCGGCGC AATCGCCATT TTAATCCTCG CCTGGCTCAT CAGTACGGTT 1051 GTCGGCGAAA TGCACACGGG CGACTACCTC TCCACGCTGG TTGCGGGCAA 1101 CATCCATCCC GGCTTCCTGC CCGTCATCCT CTTCCTGCTC GCCAGCGTGA 1151 TGGCGTTTGC CACAGGCACA AGCTGGGGGA CGTTCGGCAT TATGCTGCCG 1201 ATTGCCGCCG CCATGGCGGT CAAAGTCGAA CCCGCGCTGA TTAtcccGTG 1251 TATGTCCGCA GTAATGGCGG GGGCGGTATG CGGCGACCAC TGTTCGCCCA 1301 TCTCCGACAC GACCATCCTG TCGTCCACCG GCGCGCGCTG CAACCACATC 1351 GACCACGTTA CCTCGCAACT GCCTTATGCC CTGACGGTTG CCGCCGCCGC 1401 CGCATCGGGC TACCTCGCAT TGGGTCTGAC AAAATCCGCG CTGTTGGGCT 1451 TTGGCACGAC CGGTATTGTA TTGGCGGTGC TGATTTTTCT GTTGAAAGAT 1501 AAAAAACGCG CCGACGTTTG A This encodes a protein having amino acid sequence <SEQ LD 696>: 1 MQLIDYSHSF FSWPPFLAL ALAVITRRVL LSLGIGILVG VAFLVGGNPV 51 DGLTHLKDMV VGLAWADGDW SLGKPKILVF LILLGIFTSL LTYSGSNQAF 101 ADWAKRHIKN RCGAKMLTAC LVFVTFIDDY FHSLAVGAIA RPVTDKFKVS 151 RAKLAYILDS TASPMCVLMP VSSWGASIIA TLAGLLVTYK ITEYTPMGTF 201 VAMSLMNYYA LFALIMVFW AWFSFDIGSM ARFEQAALNE AQDETAASDA 251 TKGRVYALII PVLALIASTV SAMIYTGAQA SETFSILGAF ENTDVNTSLV 301 FGGTCGVLAV VLCTFGTIKT ADYPKAVWQG AKSMFGAIAI LILAWLISTV 351 VGEMHTGDYL STLVAGNIHP GFLPVILFLL ASVMAFATGT SWGTFGIMLP 401 lAAAMAVKVE PALIIPCMSA VMAGAVCGDH CSPISDTTIL SSTGARCNHI 451 DHVTSQLPYA LTVAAAAASG YLALGLTKSA LLGFGTTGIV LAVLIFLLKD 501 KKRADV* ORF26ng and ORF26-1 show 98.4% identity in 505 aa overlap: 10 20 30 40 50 60 orf26-1. pep MQLIDYSHSFFSWPPFLALALAVITRRVLLSLGIGILVGVAFLVGGNPVDGLTHLKDMV II I I I II I II II I I I I I I II I I I II I II II II I II I I I I I II II II I I I I II II I I I II I orf26ng MQLIDYSHSFFSWPPFLALALAVITRRVLLSLGIGILVGVAFLVGGNPVDGLTHLKDMV 10 20 30 40 50 60 70 80 90 100 110 120 orf26-1. pep VGLAWSDGDWSLGKPKILVFLILLGIFTSLLTYSGSNQAFADWAKRHIKNRRGAKMLTAC I I II I : I I I I I I I I I I I I II I I I II I II II II I I I II I I I II I I I I I I I I I II II I I I I orf26ng VGLAWADGDWSLGKPKILVFLILLGIFTSLLTYSGSNQAFADWAKRHIKNRCGAKMLTAC 70 80 90 100 110 120 130 140 150 160 170 180 orf26-1. pep LVFVTFIDDYFHSLAVGAIARPVTDKFKVSRTKLAYILDSTAAPMCVLMPVSSWGASIIA I II I I I II I I I I II I I I I II I I I II I II II I : I I I II II I I I : II I I I I II I I I I I II I I orf26ng LVFVTFIDDYFHSLAVGAIARPVTDKFKVSRAKLAYILDSTASPMCVLMPVSSWGASIIA 130 140 150 160 170 180 190 200 210 220 230 240 orf26-1. pep TLAGLLVTYKITEYTPMGTFVAMSLMNYYALFALIMVFWAWFSFDIGSMARFEQAALNE I I I I I II I II I II II I I I I I I I II I I I I I I I I I II II II I I I I I I I I I I II I I I I I II I I orf26ng TLAGLLVTYKITEYTPMGTFVAMSLMNYYALFALIMVFWAWFSFDIGSMARFEQAALNE 190 200 210 220 230 240 250 260 270 280 290 300 orf26-1. pep AHDETAVSDATKGRVYALIIPVLALIASTVSAMIYTGAQASETFSILGAFENTDVNTSLV I : II II : I I I II I II I II I I I II I I II I I I I I I II II I I I II I I I I I I I II I I I I I I I I I orf26ng AQDETAASDATKGRVYALIIPVLALIASTVSAMIYTGAQASETFSILGAFENTDVNTSLV 250 260 270 280 290 300 310 320 330 340 350 360 orf26-1. pep FGGTCGVLAWLCTLGTIKTADYPKAVWQGAKSMFGAIAILILAWLISTWGEMHTGDYL I I I I I II I I II II I : II I II I I II II II II I II I I I II I II I I II I I I I I II II II I II I orf26ng FGGTCGVLAVVLCTFGTIKTADYPKAVWQGAKSMFGAIAILILAWLISTWGEMHTGDYL 310 320 330 340 350 360 370 380 390 400 410 420 orf26-1.pep STLVAGNIHPGFLPVILFLLASVMAFATGTSWGTFGIMLPIAAAMAVKVEPALIIPCMSA I I I I I I I I I II I II I I I I II II I I I I I II I I I I I I I I I I I I I II I I I I I II I I I I I I I I I orf26ng STLVAGNIHPGFLPVILFLLASVMAFATGTSWGTFGIMLPIAAAMAVKVEPALIIPCMSA 370 380 390 400 410 420 430 440 450 460 470 480 orf26-1. pep VMAGAVCGDHCSPISDTTILSSTGARCNHIDHVTSQLPYALTVAAAAASGYLALGLTKSA II I I I II I I II I II I I II I I II I I I II II I II I I I I II I II II II I II I I II II II I I II orf26ng VMAGAVCGDHCSPISDTTILSSTGARCNHIDHVTSQLPYALTVAAAAASGYLALGLTKSA 430 440 450 460 470 480 490 500 orf26-1. pep LLGFGTTGIVLAVLIFLLKDKKRANAX I I I I II I I II I II I I I II I I I I I I :: orf26ng LLGFGTTGIVLAVLIFLLKDKKRADVX 490 500 In addition, ORF26 ng shows significant homology to a hypothetical H.influenzae protein: sp|P44263|YF86_HAEIN HYPOTHETICAL PROTEIN HI1586 >gi I 1074850 | pir II C64037 hypothetical protein HI1586 - Haemophilus influenzae (strain Rd KW20) >gi 11574427 (U32832) H. influenzae predicted coding region HI1586 [Haemophilus influenzae] Length = 519 Score = 538 bits (1370), Expect = e-152 Identities = 280/507 (55%), Positives = 346/507 (68%), Gaps = 7/507 (1%) Query: 1 MQLIDYSHSFFS PPFLALALAVITRRXXXXXXXXXXXXXAFLVGGNPVDGLTHLKDMV 60 M+LID+S S +S+VP LA+ LA+ TRR L +L V Sbjct: 14 MELIDFSSSVWSIVPALLAIILAIATRRVLVSLSAGIIIGSLMLSDWQIGSAFNYLVKNV 73 Query: 61 VGLAWADGDWSLGKPKILVFLILLGIFTSLLTYSGSNQAFADWAKRHIKNRCGAKMLTAC 120 V L +ADG+ + I++FL+LLG+ T+LLT SGSN+AFA+WA+ IK R GAK+L A Sbjct: 74 VSLVYADGEIN-SNMNIVLFLLLLGVLTALLTVSGSNRAFAEWAQSRIKGRRGAKLLAAS 132 Query: 121 LVFVTFIDDYFHSLAVGAIARPVTDKFKVSRAKLAYILDSTASPMCVLMPVSSWGASIIA 180 LVFVTFIDDYFHSLAVGAIARPVTD+FKVSRAKLAYILDSTA+PMCV+MPVSSWGA II Sbjct: 133 LVFVTFIDDYFHSLAVGAIARPVTDRFKVSRAKLAYILDSTAAPMCVMMPVSSWGAYIIT 192 Query: 181 TLAGLLVTYKITEYTPMGTFVAMSLMNYYALFALIMVFWAWFSFDIGSMARFEQAALNE 240 + GLL TY ITEYTP+G FVAMS MN+YA+F++IMVF VA+FSFDI SM R E+ AL Sbjct: 193 LIGGLLATYSITEYTPIGAFVAMSSMNFYAIFSIIMVFFVAYFSFDIASMVRHEKLALKN 252 Query: 241 AQDETAASDATKGRVYALIIPVLALIASTVSAMIYTGAQA SETFSILGAFENTDVN 296 +D+ TKG+V LI+P+L LI +TVS MIYTGA+A + FS+LG FENT V Sbjct: 253 TEDQLEEETGTKGQVRNLILPILVLIIATVSMMIYTGAEALAADGKVFSVLGTFENTWG 312 Query: 297 TSLVFGGTCGVL—AVVLCTFGTIKTADYPKAVWQGAKSMFGXXXXXXXXXXXSTWGEM 354 TSLV GG C ++ +++ + +Y ++ G KSM G + +VG+M Sbjct: 313 TSLWGGFCSIIISTLLIILDRQVSVPEYVRSWIVGIKSMSGAIAILFFAWTINKIVGDM 372 Query: 355 HTGDYLSTLVAGNIHPGFLPVILFLLASVMAFATGTSWGTFGIMLPIAAAMAVKVEPALI 414 TG YLS+LV+GNI FLPVILF+L + MAF+TGTSWGTFGIMLPIAAAMA P L+ Sbjct: 373 QTGKYLSSLVSGNIPMQFLPVILFVLGAAMAFSTGTSWGTFGIMLPIAAAMAANAAPELL 432 Query: 415 IPCMSAVMAGAVCGDHCSPISDTTILSSTGARCNHIDHVTSQXXXXXXXXXXXXXXXXXX 474 +PC+SAVMAGAVCGDHCSP+SDTTILSSTGA+CNHIDHVT+Q Sbjct: 433 LPCLSAVMAGAVCGDHCSPVSDTTILSSTGAKCNHIDHVTTQLPYAATVATATSIGYIW 492 Query: 475 XXXKSALLGFGTTGIVLAVLIFLLKDK 501 S L GF T + L V+IF +K + Sbjct: 493 GFTYSGLAGFAATAVSLIVIIFAVKKR 519 Based on this analysis, it is predicted that these proteins from N. meningitidis and N.gonorrhoeae, and their epitopes, could be useful antigens for vaccines or diagnostics, or for raising antibodies. Example 83 The following partial DNA sequence was identified in N. meningitidis <SEQ ID 697>: 1 ..AAGCAATGGT ATGCCGACGN .AGTATCAAG ACGGAAATGG TTATGGTCAA 51 CGATGAGCCT GCCAAAATTC TGACTTGGGA TGAAAGCGGC CGATTACTCT 101 CGGAACTGTC TATCCGCCAC CATCAACGCA ACGGGGTGGT TTTGGAGTGG 151 TATGAAGATG GTTCTAAAAA GAGCGAAGT . GTTTATCAGG ATGACAAGTT 201 GGTCAGGAAA ACCCAGTGGG ATAAGGATGG TTATTTAATC GAACCCTGA This coπesponds to the amino acid sequence <SEQ ID 698; ORF27>: 1 ..KQWYADXSIK TEMVMVNDEP AKILTWDESG RLLSELSIRH HQRNGWLEW 51 YEDGSKKSEX VYQDDKLVRK TQWDKDGYLI EP* Further work revealed the complete nucleotide sequence <SEQ ID 699>: 1 ATGAAAAAAT TATCTCGGAT TGTATTTTCA ACTGTCCTGT TGGGTTTTTC 51 GGCCGCTTTG CCGGCGCAGA CCTATTCTGT TTATTTTAAT CAGAACGGAA 101 AGCTGACGGC GACGATGTCT TCTGCCGCTT ATATCAGGCA ATATAGTGTG 151 GTGGCGGGTA TTGCGCACGC GCAGGATTTT TATTATCCGT CGATGAAGAA 201 ATATTCTGAA CCTTATATCG TTGCTTCAAC GCAAATCAAA TCTTTTGTGC 251 CTACCCTGCA AAACGGTATG TTGATTTTGT GGCATTTTAA TGGTCAGAAA 301 AAAATGGCGG GGGGCTTCAG CAAGGGTAAG CCGGACGGGG AGTGGGTCAA 351 CTGGTATCCG AACGGTAAAA AATCTGCCGT TATGCCTTAT AAAAATGGCT 401 TGAGTGAGGG TACGGGATAC CGCTATTACC GTAACGGCGG CAAGGAAAGC 451 GAAATCCAGT TTAAGCAAAA TAAGGCAAAC GGCGTATGGA AGCAATGGTA 501 TGCCGACGGC AGTATCAAGA CGGAAATGGT TATGGTCAAC GATGAGCCTG 551 CCAAAATTCT GACTTGGGAT GAAAGCGGCC GATTACTCTC GGAACTGTCT 601 ATCCGCCACC ATCAACGCAA CGGGGTGGTT TTGGAGTGGT ATGAAGATGG 651 TTCTAAAAAG AGCGAAGCTG TTTATCAGGA TGACAAGTTG GTCAGGAAAA 701 CCCAGTGGGA TAAGGATGGT TATTTAATCG AACCCTGA This coπesponds to the .amino acid sequence <SEQ ID 700; ORF27-l>: 1 MKKLSRIVFS TVLLGFSAAL PAQTYSVYFN QNGKLTATMS SAAYIRQYSV 51 VAGIAHAQDF YYPSMKKYSE PYIVASTQIK SFVPTLQNGM LILWHFNGQK 101 KMAGGFSKGK PDGEWVNWYP NGKKSAVMPY KNGLSEGTGY RYYRNGGKES 151 EIQFKQNKAN GVWKQWYADG SIKTEMVMVN DEPAKILTWD ESGRLLSELS 201 IRHHQRNGW LEWYEDGSKK SEAVYQDDKL VRKTQWDKDG YLIEP* Computer analysis of this amino acid sequence gave the following results: Homology with a predicted ORF from N.meninsitidis (sfrain Al ORF27 shows 91.5% identity over a 82aa overlap with an ORF (ORF27a) from sfrain A of N. meningitidis: 10 20 30 orf27.pep KQWYADXSIKTEMVMVNDEPAKILTWDESG I I I II I : I I I II II I I I I I I I I II I I I I I orf27a LSEGTGXRYYRNGGKESEIQFKQNKANGVWKQWYADGNIKTEMVMVNDEPAKILTWDESG 140 150 160 170 180 190 40 50 60 70 80 orf27. pep RLLSELSIRHHQRNGWLEWYEDGSKKSEXVYQDDKLVRKTQWDKDGYLIEPX II II I I I I : I I I I II II II I I I I I II I II I II I I II I II II II I I I I II orf27a RLLSELSIHHHXRNGWLEWYEDGSKKXEAVYQDDKLVRKTQWDXDGYLIEPX 200 210 220 230 240 The complete length ORF27a nucleotide sequence <SEQ LD 701> is: 1 ATGAAAAAAT TATCTCGGAT TGTATTTTCA ACTGTCCTGT TGGGTTTTTC 51 GGCCGCTTTG CCGGCGCAGA NCTATTCTGT TTATTTTAAT CAGAACGGGA 101 AACTGACGGC GACGNTGTCT TCTGCCGCNT ATATCAGGCA ATATAGTGTG 151 GCGGAGGGTA TTGCGCACGC GCAGGANTTT TANTATCCGT CGATGAAGAA 201 ATATTCCGAA CCTTATATCG TTGCTTCAAC GCAAATCAAA TCTTTTGTGC 251 CTACCCTGCA AAACGGTATG TTGATTTTGT GGCATTTTAA NGGTCAGAAA 301 AAAATGGCNG GGGGCTTCAG CAAGGGTAAG CCGGACGGGG AGTGGGTCAA 351 CTGGTATCCG AACGGTAAAA AATCTGCCGT TATGCCTTAT AAAAATGGTT 401 TGAGTGAAGG TACGGGGTNN CGCTATTACC GTAACGGCGG CAAGGAAAGC 451 GAAATCCAGT TTAAACAGAA TAAGGCAAAC GGCGTATGGA AGCAATGGTA 501 TGCCGACGGC AATATCAAAA CGGAAATGGT TATGGTCAAT GATGAGCCTG 551 CCAAAATTCT GACATGGGAT GAAAGCGGTC GATTACTCTC GGAACTGTCT 601 ATCCATCATC ATNAACGTAA TGGAGTAGTC TTAGAGTGGT ATGAAGATGG 651 TTCTAAAAAG ANTGAAGCTG TTTATCAGGA TGATAAGTTG GTCAGGAAAA 701 CCCAGTGGGA TAANGATGGT TATTTAATCG AACCCTGA This encodes a protein having amino acid sequence <SEQ ID 702>: 1 MKKLSRIVFS TVLLGFSAAL PAQXYSVYFN QNGKLTATXS SAAYIRQYSV 51 AEGIAHAQXF XYPSMKKYSE PYIVASTQIK SFVPTLQNGM LILWHFXGQK 101 KMAGGFSKGK PDGEWVNWYP NGKKSAVMPY KNGLSEGTGX RYYRNGGKES 151 EIQFKQNKAN GVWKQWYADG NIKTEMVMVN DEPAKILTWD ESGRLLSELS 201 IHHHXRNGW LEWYEDGSKK XEAVYQDDKL VRKTQWDXDG YLIEP* ORF27a and ORF27-1 show 94.7% identity in 245 aa overlap: 10 20 30 40 50 60 orf27a. pep MKKLSRIVFSTVLLGFSAALPAQXYSVYFNQNGKLTATXSSAAYIRQYSVAEGIAHAQXF orf27-1 MKKLSRIVFSTVLLGFSAALPAQTYSVYFNQNGKLTATMSSAAYIRQYSWAGIAHAQDF 10 20 30 40 50 60 70 80 90 100 110 120 orf27a. pep XYPSMKKYSEPYIVAΞTQIKSFVPTLQNGMLILWHFXGQKKMAGGFSKGKPDGEWVNWYP I I I I II I I I I I I I I I I I I II I II I II II II II I II I I I II I I I I II I I I II II I II I I orf27-1 YYPSMKKYSEPYIVASTQIKSFVPTLQNGMLILWHFNGQKKMAGGFSKGKPDGEWVNWYP 70 80 90 100 110 120 130 140 150 160 170 180 orf27a . pep NGKKSAVMPYKNGLSEGTGXRYYRNGGKESEIQFKQNKANGVWKQWYADGNIKTEMVMVN I I I I I II I I I I I I I I I I I I II I II I I I I I I I II I II II II I I I II I I II : I I I I I I II I orf27-1 NGKKSAVMPYKNGLSEGTGYRYYRNGGKESEIQFKQNKANGVWKQWYADGSIKTEMVMVN 130 140 150 160 170 180 190 200 210 220 230 240 orf27a . pep DEPAKILTWDESGRLLSELSIHHHXRNGWLEWYEDGSKKXEAVYQDDKLVRKTQWDXDG I II I I I I II II I I I I I I I I I I : I I I I I II II I II I II II I I II I II I I I II II II II orf27-1 DEPAKILTWDESGRLLSELSIRHHQRNGWLEWYEDGSKKSEAVYQDDKLVRKTQWDKDG 190 200 210 220 230 240 orf27a. pep YLIEPX I II I II orf27-l YLIEPX Homology with a predicted ORF from N.sonorrhoeae ORF27 shows 96.3% identity over 82 aa overlap with a predicted ORF (ORF27ng) from N.gonorrhoeae: orf27.pep KQWYADXSIKTEMVMVNDEPAKILTWDESG 30 I II I II I I I I I I II I I I I I I I I II II II I orf27ng LSEGTGYRYYRNGGKESEIQFKQNKANGVWKQWYADGSIKTEMVMVNDEPAKILTWDESG 193 orf27.pep RLLSELSIRHHQRNGWLEWYEDGSKKSEXVYQDDKLVRKTQWDKDGYLIEP 82 II I II II I II I : I I I I I I I II I I II I II I I I I I II I I I I I I I I I II I I II I orf27ng RLLSELSIRHHKRNGWLEWYEDGSKKSEAVYQDDKLVRKTQWDKDGYLIEP 245 The complete length ORF27ng nucleotide sequence <SEQ ED 703> is: 1 ATGAAGAAAT TATCTCGGAT TGTATTTTCA ATCGTACTGT TGGGTTTTTC 51 GGCCGCTTTG CCGGCGCAGA CCTATTCTGT TTATTTTAAT CAGAACGGGA 101 AACTGACGGC GACGATGTCT TCTGCCGCTT ATATCAGGCA ATATAGTGTG 151 GCGGCGGGTA TCGCACACGC GCAGGATTTT TATTATCCGT CGATGAAGAA 201 ATATTCCGAA CCTTATATCG TTGCTTCAAC GCAAATCAAA TCTTTTGTGC 251 CTACCCTGCA AAACGGTATG TTGATTTTGT GGCATTTTAA TGGTCAGAAA 301 AAAATGGCGG GGGGCTTCAG CAAGGGTAAG CCGGACGGGG AATGGGTCAA 351 CTGGTATCCG AACGGTAAAA AATCTGCGGT TATGCCTTAT AAAAATGGCT 401 TGAGTGAGGG TACGGGATAC CGTTATTACC GTAACGGCGG CAAGGAAAGC 451 GAAATCCAGT TTAAGCAAAA TAAGGCGAAC GGCGTATGGA AGCAATGGTA 501 TGCCGATGGA AGTATCAAGA CGGAAATGGT TATGGTCAAC GATGAGCCTG 551 CCAAAATTCT GACTTGGGAT GAAAGCGGCC GATTACTTTC GGAACTGTCT 601 ATCCGCCACC ATAAACGCAA CGGGGTGGTT TTGGAGTGGT ATGAAGATGG 651 TTCTAAAAAG AGCGAGGCTG TTTATCAGGA TGACAAGTTG GTCAGGAAAA 701 CCCAATGGGA TAAGGATGGT TATTTAATCG AACCCTGA This encodes a protein having amino acid sequence <SEQ LD 704>: 1 MKKLSRIVFS TVLLGFSAAL PAQTYSVYFN QNGKLTATMS SAAYIRQYSV 51 AAGIAHAQDF YYPSMKKYSE PYIVASTQIK SFVPTLQNGM LILWHFNGQK 101 KMAGGFSKGK PDGEWVNWYP NGKKSAVMPY KNGLSEGTGY RYYRNGGKES 151 EIQFKQNKAN GVWKQWYADG SIKTEMVMVN DEPAKILTWD ESGRLLSELS 201 IRHHKRNGW LEWYEDGSKK SEAVYQDDKL VRKTQWDKDG YLIEP* ORF27ng and ORF27-1 show 98.8% identity in 245 aa overlap: 10 20 30 40 50 60 orf27-1. pep MKKLSRIVFSTVLLGFSAALPAQTYSVYFNQNGKLTATMSSAAYIRQYSWAGIAHAQDF I II I II I II I II II II I I I II I II II I II I II II I I I I I I I I I I I I I I I : I I I I I II II orf27ng MKKLSRIVFSIVLLGFSAALPAQTYSVYFNQNGKLTATMSSAAYIRQYSVAAGIAHAQDF 10 20 30 40 50 60 70 80 90 100 110 120 orf27-1. pep YYPSMKKYSEPYIVASTQIKSFVPTLQNGMLILWHFNGQKKMAGGFSKGKPDGEWVNWYP I I I II I II II II I II II II I II I II I I I II II I II I I I I I I I I I II I I I I II II I I I I II orf27ng YYPSMKKYSEPYIVASTQIKSFVPTLQNGMLILWHFNGQKKMAGGFSKGKPDGEWVNWYP 70 80 90 100 110 120 130 140 150 160 170 180 orf27-1. pep NGKKSAVMPYKNGLSEGTGYRYYRNGGKESEIQFKQNKANGVWKQWYADGSIKTEMVMVN I II II I II I I I I I II I I I I I I I I II I I I I II I II II I II I I I II I I II I II I I I I I II II orf27ng NGKKSAVMPYKNGLSEGTGYRYYRNGGKESEIQFKQNKANGVWKQWYADGSIKTEMVMVN 130 140 150 160 170 180 190 200 210 220 230 240 orf27-1. pep DEPAKILTWDESGRLLSELSIRHHQRNGWLEWYEDGSKKSEAVYQDDKLVRKTQWDKDG I I II I II I I I I I I I I II II I II I I : II I I II I II II I II I I I II I I I I I II I I I I I I II I orf27ng DEPAKILTWDESGRLLSELSIRHHKRNGWLEWYEDGSKKSEAVYQDDKLVRKTQWDKDG 190 200 210 220 230 240 orf27-l.pep YLIEPX I orf27ng YLIEPX Based on this analysis, including the putative leader sequence in the gonococcal protein, it was predicted that the proteins from N. meningitidis and N.gonorrhoeae, and their epitopes, could be useful antigens for vaccines or diagnostics, or for raising antibodies. ORF27-1 (24.5kDa) was cloned in pET and pGex vectors and expressed in E.coli, as described above. The products of protein expression and purification were analyzed by SDS-PAGE. Figure 17A shows the results of affinity purification of the GST-fusion protein, and Figure 17B shows the results of expression of the His-fusion in E.coli. Purified GST-fusion protein was used to immunise mice, whose sera were used for ELISA, which gave a positive result, confirming that ORF27-1 is a surface-exposed protein and a useful immunogen. Example 84 The following partial DNA sequence was identified in N. meningitidis <SEQ ID 705>: 1 ATGAAATTTA CCAAGCACCC CGTCTGGGCA ATGGCGTTCC GCCCATTTTA 51 TTCGCTGGCG GCTCTGTACG GCGCATTGTC CGTATTGCTG TGGGGTTTCG 101 GCTACACGGG AACGCACkAG CTGTCCGGTT TCTATTGGCA CGCGCATGAg 151 ATGATTTGGG GTTATGCCGG ACTGGTCGTC ATCGCCTTCC TGCTGACCGC 201 CGTCGCCACT TGGACGGGGC AGCCGCCCAC GCGGGGCGGC GTaTCTGGTC 251 GGCTTGACTA TCTTTTGGCT GGCTGCGCGG ATTGCCGCCT TTATCCCGGG 301 TTGGGGTGCG TCGGCAAGCG GCATACTCGG TACGCTGTTT TTCTGGTACG 351 GCGCGGTGTG CATGGCTTTG CCCGTTATCC GTTCGCAGAA TCAACGCAAC 401 TATGTTgCCG TGTTCGCGCT GTTCGTCTTG GGCGGCACGC ATGCGGCGTT 451 CCACGTCCAG CTGCACAACG GCAACCTAGG CGGACTCTTG AGCGGATTGC 501 AGTCGGGCTT GGTGATG This coπesponds to the .amino acid sequence <SEQ LD 706; ORF47>: 1 MKFTKHPVWA MAFRPFYSLA ALYGALSVLL WGFGYTGTHX LSGFYWHAHE 51 MIWGYAGLW IAFLLTAVAT WTGQPPTRGG VLVGLTIFWL AARIAAFIPG 101 WGASASGILG TLFFWYGAVC MALPVIRSQN QRNYVAVFAL FVLGGTHAAF 151 HVQLHNGNLG GLLSGLQSGL VM Further work revealed the complete nucleotide sequence <SEQ ID 707>: 1 ATGAAATTTA CCAAGCACCC CGTCTGGGCA ATGGCGTTCC GCCCATTTTA 51 TTCGCTGGCG GCTCTGTACG GCGCATTGTC CGTATTGCTG TGGGGTTTCG 101 GCTACACGGG AACGCACGAG CTGTCCGGTT TCTATTGGCA CGCGCATGAG 151 ATGATTTGGG GTTATGCCGG ACTGGTCGTC ATCGCCTTCC TGCTGACCGC 201 CGTCGCCACT TGGACGGGGC AGCCGCCCAC GCGGGGCGGC GTTCTGGTCG 251 GCTTGACTAT CTTTTGGCTG GCTGCGCGGA TTGCCGCCTT TATCCCGGGT 301 TGGGGTGCGT CGGCAAGCGG CATACTCGGT ACGCTGTTTT TCTGGTACGG 351 CGCGGTGTGC ATGGCTTTGC CCGTTATCCG TTCGCAGAAT CAACGCAACT 401 ATGTTGCCGT GTTCGCGCTG TTCGTCTTGG GCGGCACGCA TGCGGCGTTC 451 CACGTCCAGC TGCACAACGG CAACCTAGGC GGACTCTTGA GCGGATTGCA 501 GTCGGGCTTG GTGATGGTGT CGGGTTTTAT CGGTCTGATT GGTACGCGGA 551 TTATTTCGTT TTTTACGTCC AAACGCTTGA ATGTGCCGCA GATTCCCAGT 601 CCGAAATGGG TGGCGCAGGC TTCGCTGTGG CTGCCCATGC TGACTGCCAT 651 GCTGATGGCG CACGGTGTGT TGGCTTGGCT GTCTGCCGTT TTTGCCTTTG 701 CGGCAGGTGT GATTTTTACC GTGCAGGTGT ACCGCTGGTG GTATAAACCC 751 GTGTTGAAAG AGCCGATGCT GTGGATTCTG TTTGCCGGCT ATCTGTTTAC 801 CGGATTGGGG CTGATTGCGG TCGGCGCGTC TTATTTCAAA CCCGCTTTCC 851 TCAATCTGGG TGTGCATCTG ATCGGGGTCG GCGGTATCGG CGTGCTGACT 901 TTGGGCATGA TGGCGCGTAC CGCGCTTGGT CATACGGGCA ATCCGATTTA 951 TCCGCCGCCC AAAGCCGTTC CCGTTGCGTT TTGGCTGATG ATGGCGGCAA 1001 CCGCCGTCCG TATGGTTGCC GTATTTTCTT CCGGCACTGC CTACACGCAC 1051 AGCATCCGCA CCTCTTCGGT TTTGTTTGCA CTCGCGCTTT TGGTGTATGC 1101 GTGGAAGTAT ATTCCTTGGC TGATTCGTCC GCGTTCGGAC GGCAGGCCCG 1151 GTTGA This coπesponds to the amino acid sequence <SEQ ID 708; ORF47-l>: 1 MKFTKHPVWA MAFRPFYSLA ALYGALSVLL WGFGYTGTHE LSGFYWHAHE 51 MIWGYAGLW IAFLLTAVAT WTGQPPTRGG VLVGLTIFWL AARIAAFIPG 101 WGASASGILG TLFFWYGAVC MALPVIRSQN QRNYVAVFAL FVLGGTHAAF 151 HVQLHNGNLG GLLSGLQSGL VMVSGFIGLI GTRIISFFTS KRLNVPQIPS 201 PKWVAQASLW LPMLTAMLMA HGVLAWLSAV FAFAAGVIFT VQVYRWWYKP 251 VLKEPMLWIL FAGYLFTGLG LIAVGASYFK PAFLNLGVHL IGVGGIGVLT 301 LGMMARTALG HTGNPIYPPP KAVPVAFWLM MAATAVRMVA VFSSGTAYTH 351 SIRTΞSVLFA LALLVYAWKY IPWLIRPRSD GRPG* Computer analysis of this amino acid sequence predicts a leader peptide and also gave the following results: Homology with a predicted ORF from N. meningitidis (sfrain A) ORF47 shows 99.4% identity over a 172aa overlap with an ORF (ORF47a) from sfrain A of N meningitidis: 10 20 30 40 50 60 orf47.pep MKFTKHPVWAMAFRPFYSLAALYGALSVLLWGFGYTGTHXLSGFYWHAHEMIWGYAGLW I I I I I II I I II II I I I I I II I I II I I II I II I I I I II II I I I I I I I I I II I I I II II II orf47a MKFTKHPVWAMAFRPFYSLAALYGALSVLLWGFGYTGTHELSGFYWHAHEMIWGYAGLW 10 20 30 40 50 60 70 80 90 100 110 120 orf47. pep IAFLLTAVATWTGQPPTRGGVLVGLTIFWLAARIAAFIPGWGASASGILGTLFFWYGAVC I I I I I II I I I II I I I I I I I I I I II I I I I I I I II I I II I I I I I I II I I I I I I I I I I II I II orf47a IAFLLTAVATWTGQPPTRGGVLVGLTIFWLAARIAAFIPGWGASASGILGTLFFWYGAVC 70 80 90 100 110 120 130 140 150 160 170 orf47.pep MALPVIRSQΝQRΝYVAVFALFVLGGTHAAFHVQLHΝGΝLGGLLSGLQSGLVM II I II I I II I I I I I I I I II I II I I I II I II I I I I I II II II II II I I I II I I orf47a MALPVIRSQΝQRΝYVAVFALFVLGGTHAAFHVQLHΝGΝLGGLLSGLQSGLVMVSGFIGLI 130 140 150 160 170 180 orf47a GTRIISFFTSKRLΝVPQIPSPKWVAQASLWLPMLTAMLMAHGVMPWLSAAFAFAAGVIFT 190 200 210 220 230 240 The complete length ORF47a nucleotide sequence <SEQ ID 709> is: 1 ATGAAATTTA CCAAGCACCC CGTTTGGGCA ATGGCGTTCC GCCCGTTTTA 51 TTCACTGGCG GCTCTGTACG GCGCATTGTC CGTATTGCTG TGGGGTTTCG 101 GCTACACGGG AACGCACGAG CTGTCCGGTT TCTATTGGCA CGCGCATGAG 151 ATGATTTGGG GTTATGCCGG ACTGGTCGTC ATCGCCTTCC TGCTGACCGC 201 CGTCGCCACT TGGACGGGGC AGCCGCCCAC GCGGGGCGGC GTTCTGGTCG 251 GCTTGACTAT CTTTTGGCTG GCTGCGCGGA TTGCCGCCTT TATCCCGGGT 301 TGGGGTGCGT CGGCAAGCGG CATACTCGGT ACGCTGTTTT TCTGGTACGG 351 CGCGGTGTGC ATGGCTTTGC CCGTTATCCG TTCGCAGAAT CAACGCAATT 401 ATGTTGCCGT GTTCGCGCTG TTCGTCTTGG GCGGTACGCA CGCGGCGTTC 451 CACGTCCAGC TGCACAACGG CAACCTAGGC GGACTCTTGA GCGGATTGCA 501 GTCGGGCTTG GTGATGGTGT CGGGTTTTAT CGGTCTGATT GGTACGCGGA 551 TTATTTCGTT TTTTACGTCC AAACGGTTGA ATGTGCCGCA GATTCCCAGT 601 CCGAAATGGG TGGCGCAGGC TTCGCTGTGG CTGCCCATGC TGACCGCCAT 651 GCTGATGGCG CACGGCGTGA TGCCTTGGCT GTCGGCGGCT TTCGCGTTTG 701 CGGCAGGTGT GATTTTTACC GTGCAGGTGT ACCGCTGGTG GTATAAGCCT 751 GTGTTGAAAG AGCCGATGCT GTGGATTCTG TTTGCCGGCT ATCTGTTTAC 801 CGGATTGGGG CTGATTGCGG TCGGCGCGTC TTATTTCAAA CCCGCTTTCC 851 TCAATCTGGG TGTGCATCTG ATCGGGGTCG GCGGTATCGG CGTGCTGACT 901 TTGGGCATGA TGGCGCGTAC CGCGCTCGGT CATACGGGCA ATCCGATTTA 951 TCCGCCGCCC AAAGCCGTTC CCGTTGCGTT TTGGCTGATG ATGGCGGCAA 1001 CCGCCGTCCG TATGGTTGCC GTATTTTCTT CCGGCACTGC CTACACGCAC 1051 AGCATACGCA CCTCTTCGGT TTTGTTTGCA CTCGCGCTTT TGGTGTATGC 1101 GTGGAAGTAT ATTCCTTGGC TGATTCGTCC GCGTTCGGAC GGCAGGCCCG 1151 GTTGA This encodes a protein having amino acid sequence <SEQ ID 710>: 1 MKFTKHPVWA MAFRPFYSLA ALYGALSVLL WGFGYTGTHE LSGFYWHAHE 51 MIWGYAGLW IAFLLTAVAT WTGQPPTRGG VLVGLTIFWL AARIAAFIPG 101 WGASASGILG TLFFWYGAVC MALPVIRSQN QRNYVAVFAL FVLGGTHAAF 151 HVQLHNGNLG GLLSGLQSGL VMVSGFIGLI GTRIISFFTS KRLNVPQIPS 201 PKWVAQASLW LPMLTAMLMA HGVMPWLSAA FAFAAGVIFT VQVYRWWYKP 251 VLKEPMLWIL FAGYLFTGLG LIAVGASYFK PAFLNLGVHL IGVGGIGVLT 301 LGMMARTALG HTGNPIYPPP KAVPVAFWLM MAATAVRMVA VFSSGTAYTH 351 SIRTSSVLFA LALLVYAWKY IPWLIRPRSD GRPG* ORF47a and ORF47-1 show 99.2% identity in 384 aa overlap: 10 20 30 40 50 60 orf47a . pep MKFTKHPVWAMAFRPFYSLAALYGALSVLLWGFGYTGTHELSGFYWHAHEMIWGYAGLW I II I I I II I II II II I I I I I I I I I II II I I II I I I I II II I II I I I I I II I II I I I I I II orf47-1 MKFTKHPVWAMAFRPFYSLAALYGALSVLLWGFGYTGTHELSGFYWHAHEMIWGYAGLW 10 20 30 40 50 60 70 80 90 100 110 120 orf47a . pep IAFLLTAVATWTGQPPTRGGVLVGLTIFWLAARIAAFIPGWGASASGILGTLFFWYGAVC I I I I I II I I I I I I I I I II I I I I I II I I I I II I II I II I I II I I I I I II I I II I I I I I II I orf47-1 IAFLLTAVATWTGQPPTRGGVLVGLTIFWLAARIAAFIPGWGASASGILGTLFFWYGAVC 70 80 90 100 110 120 130 140 150 160 170 180 orf47a . pep MALPVIRSQNQRNYVAVFALFVLGGTHAAFHVQLHNGNLGGLLSGLQSGLVMVSGFIGLI I I II I II II I I I I I I I II I I II I I I I I I I I II I II I I I I II II II I I I II I II I I I I II I orf47-1 MALPVIRSQNQRNYVAVFALFVLGGTHAAFHVQLHNGNLGGLLSGLQSGLVMVSGFIGLI 130 140 150 160 170 180 190 200 210 220 230 240 orf47a . pep GTRIISFFTSKRLNVPQIPSPKWVAQASLWLPMLTAMLMAHGVMPWLSAAFAFAAGVIFT I I II I II I I I I I I I I I I II I II I I II II I I I II I II I II I I I I : I I I I: II I II II I I I orf47-1 GTRIISFFTSKRLNVPQIPSPKWVAQASLWLPMLTAMLMAHGVLAWLSAVFAFAAGVIFT 190 200 210 220 230 240 250 260 270 280 290 300 orf47a. ep VQVYRWWYKPVLKEPMLWILFAGYLFTGLGLIAVGASYFKPAFLNLGVHLIGVGGIGVLT II I I I I I I I II I I I I I II I I I I I I I I II II I II I II I II I II I I I I I I I II I I I II II I I orf47-1 VQVYRWWYKPVLKEPMLWILFAGYLFTGLGLIAVGASYFKPAFLNLGVHLIGVGGIGVLT 250 260 270 280 290 300 310 320 330 340 350 360 orf47a . pep LGMMARTALGHTGNPIYPPPKAVPVAFWLMMAATAVRMVAVFSSGTAYTHSIRTSSVLFA II I II I II I I I I I I I I I I I II I II I II I II I II I I I I I II I I I I I I I I I I II I I II II I I orf47-1 LGMMARTALGHTGNPIYPPPKAVPVAFWLMMAATAVRMVAVFSSGTAYTHSIRTSSVLFA 310 320 330 340 350 360 370 380 orf47a.pep LALLVYAWKYIPWLIRPRSDGRPGX I II I II I II I I II I I I I I II I I II I orf47-1 LALLVYAWKYIPWLIRPRΞDGRPGX 370 380 Homology with a predicted ORF from N.gonorrhoeae ORF47 shows 97.1% identity over 172 aa overlap with a predicted ORF (ORF47ng) from N.gonorrhoeae: ORF47 MKFTKHPVWAMAFRPFYSLAALYGALSVLLWGFGYTGTHELSGFYWHAHEMIWGYAGLW 60 I I II I I II I II I II I II I I II I I I II I I II II I I I I I II I I II I II I II I I I I I I I I I I I ORF47ng MKFTKHPVWAMAFRPFYSLAALYGALSVLLWGFGYTGTHELSGFYWHAHEMIWGYAGLW 60 ORF47 IAFLLTAVATWTGQPPTRGGVLVGLTIFWLAARIAAFIPGWGASASGILGTLFFWYGAVC 120 I II I II I I II I II I II II I I I II II I II I II II II I II I I II : I II II I II I I I II I I I ORF47ng IAFLLTAVATWTGQPPTRGGVLVGLTAFWLAARIAAFIPGWGAAASGILGTLFFWYGAVC 120 ORF47 MALPVIRSQNQRNYVAVFALFVLGGTHAAFHVQLHNGNLGGLLSGLQSGLVM 172 I I I I I I I I IM II I II I I I : I II II I I II I I I II II I II II II I II I II II I ORF47ng MALPVIRSQNRRNYVAVFAIFVLGGTHAAFHVQLHNGNLGGLLSGLQSGLVMVWGFIGLI 180 The ORF47ng nucleotide sequence <SEQ ID 711> is predicted to encode a protein comprising amino acid sequence <SEQ ID 712>: 1 MKFTKHPVWA MAFRPFYSLA ALYGALSVLL WGFGYTGTHE LSGFYWHAHE 51 MIWGYAGLW IAFLLTAVAT WTGQPPTRGG VLVGLTAFWL AARIAAFIPG 101 WGAAASGILG TLFFWYGAVC MALPVIRSQN RRNYVAVFAI FVLGGTHAAF 151 HVQLHNGNLG GLLSGLQSGL VMVWGFIGLI GMKIISFFTS KRLKLPQIPS 201 PKWVAHASLW LPMLNAILMA HRVMPWLSAA FPFAAGVIFT VQVYAGGITP 251 IEETSCGSVA GICYRLGNSS G The predicted leader peptide and transmembrane domains are identical (except for an He/ Ala substitution at residue 87 and an Leu/Ile substitution at position 140) to sequences in the meningococcal protein (see also Pseudomonas stutzeri orf396, accession number e246540): TM segments in 0RF47ng INTEGRAL Likelihood = -5. .63 Transmembrane 52 - 68 INTEGRAL Likelihood = -3. .88 Transmembrane 169 - 185 INTEGRAL Likelihood = -3. .08 Transmembrane 82 - 98 INTEGRAL Likelihood = -1, .91 Transmembrane 134 - 150 INTEGRAL Likelihood = -1, .44 Transmembrane 107 - 123 INTEGRAL Likelihood = -1. .38 Transmembrane 227 - 243 Further work revealed the complete gonococcal DNA sequence <SEQ ID 713>: 1 ATGAAATTTA CCAAACATCC CGTCTGGGCA ATGGCGTTCC GCCCGTTTTA 51 TTCACTGGCG GCACTGTACG GCGCATTGTC CGTATTGCTG TGGGGTTTCG 101 GCTACACGGG AACGCACGAG CTGTCCGGTT TCTATTGGCA CGCGCATGAG 151 ATGATTTGGG GTTATGCCGG TCTCGTCGTC ATCGCCTTCC TGCTGACCGC 201 CGTCGCCACT TGGACGGGAC AGCCGCCCAC GAGGGGCGGC GTTCTGGTCG 251 GCTTGACCGC CTTTTGGCTG GCTGCGCGGA TTGCCGCCTT TATCCCGGGT 301 TGGGGTGCGG CGGCAAGCGG CATACTCGGT ACGCTGTTTT TCTGGTACGG 351 CGCGGTGTGC ATGGCTTTGC CCGTTATCCG TtcgCAAAAC CGGCGCAACT 401 ATGtcgCCGT ATTCGCAATA TTTGTGCTGG GCGGTACGCA TGCGgcgTTC 451 CACGtccAgc tGCACAACGG CAACCTAGGC GGACTCTTGA GCGGATTGCA 501 GTCGGGCCTG GTTATGGTGT CGGGCTTTAT CGGCCTGATT GGGATGAGGA 551 TTATTTCGTT TTTTACGTCC AAACGGTTGA ACGTGCCGCA GATTCCCAGT 601 CCGAAATGGG TGGCGCAGGC TTCGCTGTGG CTACCCATGC TGACCGCCAT 651 ACTGATGGCG CACGGCGTGA TGCCTTGGCT GTCGGCGGCT TTCGCGTTTG 701 CGGCGGGCGT GATTTTTACC GTACAGGTGT ACCGCTGGTG GTATAAACCC 751 GTATTGAAAG AACCGATGCT GTGGATTCTG TTTGCCGGCT ATCTGTTTAC 801 CGGATTGGGG CTGATTGCGG TCGGCGCGTC TTATTTCAAA CCTGCCTTCC 851 TCAATCTGGG CGTACATCTG ATCGGGGTCG GCGGTATCGG CGTGCTGACT 901 TTGGGCATGA TGGCGCGTAC CGCGCTCGGT CATACGGGCA ATTCGATTTA 951 TCCGCCGCCC AAAGCCGTTC CCGTTGCGTT TTGGCTGATG ATGGCGGCAA 1001 CCGCCGTCCG TATGGTTGCC GTATTTTCTT CCGGCACTGC CTACACGCAC 1051 AGCATCCGCA CGTCTTCGGT TTTGTTTGCA CTCGCGCTGC TGGTGTATGC 1101 GTGGAAATAC ATTCCGTGGC TGATCCGTCC GCGTTCGGAC GGCAGGCCCG 1151 GTTGA This encodes a protein having amino acid sequence <SEQ ID 714; ORF47ng-l > • 1 MKFTKHPVWA MAFRPFYSLA ALYGALSVLL WGFGYTGTHE LSGFYWHAHE 51 MIWGYAGLW IAFLLTAVAT WTGQPPTRGG VLVGLTAFWL AARIAAFIPG 101 WGAAASGILG TLFFWYGAVC MALPVIRSQN RRNYVAVFAI FVLGGTHAAF 151 HVQLHNGNLG GLLSGLQSGL VMVSGFIGLI GMRIISFFTS KRLNVPQIPS 201 PKWVAQASLW LPMLTAILMA HGVMPWLSAA FAFAAGVIFT VQVYRWWYKP 251 VLKEPMLWIL FAGYLFTGLG LIAVGASYFK PAFLNLGVHL IGVGGIGVLT 301 LGMMARTALG HTGNSIYPPP KAVPVAFWLM MAATAVRMVA VFSSGTAYTH 351 SIRTSSVLFA LALLVYAWKY IPWLIRPRSD GRPG* ORF47ng-l and ORF47-1 show 97.4% identity in 384 aa overlap: 10 20 30 40 50 60 orf47-1. pep MKFTKHPVWAMAFRPFYSLAALYGALSVLLWGFGYTGTHELSGFYWHAHEMIWGYAGLW II I I I II I I I I I II I I II I I I I I I II II II I I II II II I I I II II I II I I I I I I I II I I I orf47ng-l MKFTKHPVWAMAFRPFYSLAALYGALSVLLWGFGYTGTHELSGFYWHAHEMIWGYAGLW 10 20 30 40 50 60 70 80 90 100 110 120 orf 7-1. pep IAFLLTAVATWTGQPPTRGGVLVGLTIFWLAARIAAFIPGWGASASGILGTLFFWYGAVC II I I I I I I I I I I II I I II I I II I I II I I II I I I I I I I I I I I I : II I I I II I I I I I I II I orf47ng-l IAFLLTAVATWTGQPPTRGGVLVGLTAFWLAARIAAFIPGWGAAASGILGTLFFWYGAVC 70 80 90 100 110 120 130 140 150 160 170 180 orf47-1. pep MALPVIRSQNQRNYVAVFALFVLGGTHAAFHVQLHNGNLGGLLSGLQSGLVMVSGFIGLI I II I I II I IM I I I I II I M I II I I II II I II I II I I I I II I I I I I I I I I I I II II I I II orf47ng-l MALPVIRSQNRRNYVAVFAIFVLGGTHAAFHVQLHNGNLGGLLSGLQSGLVMVSGFIGLI 130 140 150 160 170 180 190 200 210 220 230 240 orf47-1. pep GTRIISFFTSKRLNVPQIPSPKWVAQASLWLPMLTAMLMAHGVLAWLSAVFAFAAGVIFT I I I I I II I I I II I II II I I II I I II I I I II I I I I I : I I I II I : I I II : I II II I I II I orf47ng-l GMRIISFFTSKRLNVPQIPSPKWVAQASLWLPMLTAILMAHGVMPWLSAAFAFAAGVIFT 190 200 210 220 230 240 250 260 270 280 290 300 orf47-1. pep VQVYRWWYKPVLKEPMLWILFAGYLFTGLGLIAVGASYFKPAFLNLGVHLIGVGGIGVLT I II I I II I I I II I I I II I I II I I I I I II I I I I I I I II I I I II I I I I I I I I I II II I I I II orf47ng-l VQVYRWWYKPVLKEPMLWILFAGYLFTGLGLIAVGASYFKPAFLNLGVHLIGVGGIGVLT 250 260 270 280 290 300 310 320 330 340 350 360 orf47-1. pep LGMMARTALGHTGNPIYPPPKAVPVAFWLMMAATAVRMVAVFSSGTAYTHSIRTSSVLFA I I I II II I II I I I I I I I I I II I I I I I II I I II I I I I I I I I II II II I I II I I I I I I II I orf47ng-l LGMMARTALGHTGNSIYPPPKAVPVAFWLMMAATAVRMVAVFSSGTAYTHSIRTSSVLFA 310 320 330 340 350 360 370 380 orf47-1.pep LALLVYAWKYIPWLIRPRSDGRPGX I I II II II I II I I I I I I II I I II II orf4 ng-l LALLVYAWKYIPWLIRPRSDGRPGX 370 380 Furthermore, ORF47ng-l shows significant homology to an ORF from Pseudomonas stutzeri: gnl|PID|e246540 (Z73914) ORF396 protein [Pseudomonas stutzeri] Length = 396 Score = 155 bits (389), Expect = 5e-37 Identities = 121/391 (30%), Positives = 169/391 (42%), Gaps = 21/391 (5%) Query: 7 PVWAMAFRPFYSLAALYGALSVLLWGFGYTGTHELSGFY WHAHEMIWGYAGLV 59 P+W +AFRPF+ +LY L++ LW +TG GF WH HEM++G+A + Sbjct: 14 PIWRLAFRPFFLAGSLYALLAIPLWVAAWTGLWP—GFQPTGGWLAWHRHEMLFGFAMAI 71 Query: 60 VIAFLLTAVATWTGQPPTRGGVLVGLTAFWLAARIAAFIPGWGAAASGILGTLFFWYGAV 119 V FLLTAV TWTGQ G LVGL A WLAAR+ ++ G AA L LF Sbjct: 72 VAGFLLTAVQTWTGQTAPSGNRLVGLAAVWLAARL-GWLFGLPAAWLAPLDLLFLVALVW 130 Query: 120 CMALPVIRSQNRRNYVAVFAIFVLGGTHAAFXXXXXXXXXXXXXXXXXXXXXMVSGFIGL 179 MA + + +RNY V + ++ G +V+ + L Sbjct: 131 MMAQMLWAVRQKRNYPIVWLSLMLGADVLILTGLLQGNDALQRQGVLAGLWLVAALMAL 190 Query: 180 IGMRIISFFTSKRLNVPQIPSP-KWVAQASLWLPMLTAILMAHGV MPWLSAAFAFA 234 IG R+I FFT + L P W+ A L + A+L A GV P L F A Sbjct: 191 IGGRVIPFFTQRGLGKVDAVKPWVWLDVALLVGTGVIALLHAFGVAMRPQPLLGLLFV-A 249 Query: 235 AGVIFTVQVYRWWYKPVLKEPMLWILFAGYLFTGLGLIAVGASYF-KPAFXXXXXXXXXX 293 GV +++ RW+ K + K +LW L L+ + + +F A Sbjct: 250 IGVGHLLRLMRWYDKGIWKVGLLWSLHVAMLWLWAAFGLALWHFGLLAQSSPSLHALSV 309 Query: 294 XXXXXXXXXMMARTALGHTGNSIYPPPKAVPVAFWLXXXXXXXXXXXXFSSGTAYTHSIR 353 M+AR LGHTG + P + AF L F S + Sbjct: 310 GSMSGLILAMIARVTLGHTGRPLQLPAGIIG-AFVL FNLGTAARVFLSVAWPVGGLW 365 Query: 354 TSSVLFALALLVYAWKYIPWLIRPRSDGRPG 384 ++V + LA +Y W+Y P L+ R DG PG Sbjct: 366 LAAVCWTLAFALYVWRYAPMLVAARVDGHPG 396 Based on this analysis, it is predicted that the proteins from N. meningitidis .and N.gonorrhoeae, and their epitopes, could be useful antigens for vaccines or diagnostics, or for raising antibodies. Example 85 The following partial DNA sequence was identified in N. meningitidis <SEQ ID 715>: 1 ..ATGCCGTCTG AAGGTTCAGA CGGCmTCGGT GyCGGGGAAy CAGAAGyGGT 51 AGCGCATGCC CAATGAGACT TCGTGGGTTT TGAAGCGGGT GTTTTCCAAG 101 CGTCCCCAGT TGTGGTAACG GTATCCGGTG TCyAArGTCA GCTTGGGyGT 151 GATGTCGAAa CCGACACCGG CGATGACACC AAGACCyAmG CTGCTGATrC 201 TGTkGCTTTC GTGATAGGsA GGTTTGyTGG kmksAsyTTG TAyrATwkkG 251 CCTssCwsTG kAGmGCCkTk CkyTGGTkkA swGrwArTAG TCGTGGTTTy 301 TkTTyyCACC GAATGAACyT GATGTTTAAC GTGTCCGTAG GCGACGCGCG 351 CGCCGATATA GGGTTTGAAT TTATCGTTGA GTTTGAAATC GTAAATGGCG 401 GACAAGCCGA GAGAAGAAAC GGCGTGGAAG CTGCCGTTTC CCTGATGTTT 451 TGTTTGGGTT TCTTTGTAGT TGTTGTTTAT CTCTTCAGTA ACTTTTTTAG 501 TAGAAGAATT ACTTTCTTTC CATTTTCTGT AACTGGCATA ATCTGCCGCT 551 ATTCTCCAGC CGCCGAAATC .. This coπesponds to the amino acid sequence <SEQ ID 716; ORF67>: 1 ..MPSEGSDGXG XGEXEXVAHA QXDFVGFEAG VFQASPVWT VSGVXXQLGX 51 DVETDTGDDT KTXAADXVAF VIGRFXGXXL YXXAXXXXAX XWXXXXSRGF 101 XXHRMNLMFN VSVGDARADI GFEFIVEFEI VNGGQAERRN GVEAAVSLMF 151 CLGFFWWY LFSNFFSRRI TFFPFSVTGI ICRYSPAAEI .. Computer analysis of this amino acid sequence gave the following results: Homology with a predicted ORF from N.sonorrhoeae ORF67 shows 51.8% identity over 199 aa overlap with a predicted ORF (ORF67ng) from N.gonorrhoeae: orf 67 . pep MPSEGSDGXGXGEXEXVAHAQXDFVGFEAG 30 I I I I I I I I I I I I I I I I I I I I I I I I orf 67ng TNFEIAVLSGMTVRVFYCARPAPVNGGRLKMPSEGSDGIGIGESEAVAHAQRGFVGFEAG 14 6 90 100 110 120 130 140 orf67.pep VFQASPVWTVSGVXXQLGXDVETDTGDDTKTXAADXVAFVIGRFXGXXLYXXAXXXXAX 90 I I I I I I I I M M I I M M : : : : : | I I I M I I I : : orf67ng VFQASPVWAVAGVQGQAGRDVYAHARHRAEAQAAAAVAFLIGVFLRMSVRINRNCCVS I 206 orf67.pep XWXXXXSRGFXXHRMNLMFNVSVGDARADIGFEFIVEFEIVNGGQAERRNGVEAAVSLMF 150 : I : M : : : I I I I I I M I I I I I I : I I I I I I I I I I I I I I I I I I I I I I I orf67ng TRVGGKSTCYFFSRIDAVSDVSVGDARTDIGFEFWEFEIVNGGQAERRNGVECAVFLMF 266 orf 67 . pep CLGFFVV WYLFSNFFSRRITFF-PFSVTGI ICRYSPAAEI 190 I I I : : M I : : I : I I I I I I I M M M orf 67ng RLLVFYVKLVAAKSFIILSFQLFYVHGIFIWPFPVTGIIRGDAPAAEWADRHPGVDGM 326 The ORF67ng nucleotide sequence <SEQ ID 717> is predicted to encode a protein comprising amino acid sequence <SEQ ID 718>: 1 MPSETVGSIV NVGVDEΞVGF SPPFPSIQHF YRFHRIHRIR LFRPPGPMQL 51 NRHSHGSGNL GRGVWATVLS DKFPCGQVRI PACAGMTNFE IAVLSGMTVR 101 VFYCARPAPV NGGRLKMPSE GSDGIGIGES EAVAHAQRGF VGFEAGVFQA 151 SPVWAVAGV QGQAGRDVYA HARHRAEAQA_ AAAVAFLIGV FLRMSVRINR 201 NCCVSITRVG GKSTCYFFSR IDAVSDVSVG DARTDIGFEF WEFEIVNGG 251 QAERRNGVEC_ AVFLMFRLLV FYVKLVAAKS FIILSFQLFY VHGIFIWPF 301 PVTGIIRGDA PAAEWADRH PGVDGMRTDV SEIIAYRAYF VFAWSGWFRI 351 IVGNAFGGVG Based on the presence of a several putative fransmembrane domains in the gonococcal protein, it is predicted that the proteins from N. meningitidis .and N.gonorrhoeae, .and their epitopes, could be useful .antigens for vaccines or diagnostics, or for raising antibodies. Example 86 The following partial DNA sequence was identified in N. meningitidis <SEQ ID 719> 1 ATGTTTGCTT TTTTAGAAGC CTTTTTTGTC GAATACGGTT ATGCGGCTGT 51 TTTTTTTGTA TTGGTCATCT GCGGTTTCGG CGTGCCGATT CCCGAGGATT 101 TGACCTTGGT AACAGGCGGC GTGATTTCGG GTATGGGTTA TACCAATCCG 151 CATATTATGT TTGCAGTCGG TATGCTCGGC GTATTGGTCG GGGACGGCAT 201 CATGTTCGCC GCCGGACGAA TTTGGGGGCA GArArTCCTA rGGTTCArAC 251 CTATTGCGsG CATCATGACG CCGrAACGTT ATGAGCAGGT TCAGGAAAAA 301 TTCGACAAAT ACGGTAACTG GGTCTTATTT GTCGCCCGTT TCCTGCCCGG 351 TTTGAGAACG GCCGTATTTG TTACAGCCGG TATCAGCCGC AAGGTTTCAT 401 ACTTGCGTTT TATCATTATG GATGGACTGG CCGCA... This coπesponds to the amino acid sequence <SEQ ID 720; ORF78>: 1 MFAFLEAFFV EYGYAAVFFV LVICGFGVPI PEDLTLVTGG VISGMGYTNP 51 HIMFAVGMLG VLVGDGIMFA AGRIWGQXXL XFXPIAXIMT PXRYEQVQEK 101 FDKYGNWVLF VARFLPGLRT AVFVTAGISR KVSYLRFIIM DGLAA... Further work revealed the complete nucleotide sequence <SEQ ID 721>: 1 ATGTTTGCTT TTTTAGAAGC CTTTTTTGTC GAATACGGTT ATGCGGCTGT 51 TTTTTTTGTA TTGGTCATCT GCGGTTTCGG CGTGCCGATT CCCGAGGATT 101 TGACCTTGGT AACAGGCGGC GTGATTTCGG GTATGGGTTA TACCAATCCG 151 CATATTATGT TTGCAGTCGG TATGCTCGGC GTATTGGTCG GGGACGGCAT 201 CATGTTCGCC GCCGGACGAA TTTGGGGGCA GAAAATCCTA AGGTTCAAAC 251 CTATTGCGCG CATCATGACG CCGAAACGTT ATGAGCAGGT TCAGGAAAAA 301 TTCGACAAAT ACGGTAACTG GGTCTTATTT GTCGCCCGTT TCCTGCCCGG 351 TTTGAGAACG GCCGTATTTG TTACAGCCGG TATCAGCCGC AAGGTTTCAT 401 ACTTGCGTTT TATCATTATG GATGGACTGG CCGCACTGAT TTCCGTCCCT 451 ATTTGGATTT ATCTGGGCGA ATACGGTGCG CACAACATCG ATTGGCTGAT 501 GGCGAAAATG CACAGCCTGC AATCGGGTAT TTTTGTTATC TTGGGTATAG 551 GTGCGACCGT TGTCGCTTGG ATTTGGTGGA AAAAACGCCA ACGTATCCAG 601 TTTTACCGCA GCAAATTGAA AGAAAAGCGG GCGCAACGCA AAGCCGCCAA 651 GGCAGCCAAA AAAGCCGCGC AAAGCAAACA ATAA This coπesponds to the amino acid sequence <SEQ ID 722; ORF78-1 > • 1 MFAFLEAFFV EYGYAAVFFV LVICGFGVPI PEDLTLVTGG VISGMGYTNP 51 HIMFAVGMLG VLVGDGIMFA AGRIWGQKIL RFKPIARIMT PKRYEQVQEK 101 FDKYGNWVLF VARFLPGLRT AVFVTAGISR KVSYLRFIIM DGLAALISVP 151 IWIYLGEYGA HNIDWLMAKM HSLQSGIFVI LGIGATWAW IWWKKRQRIQ 201 FYRSKLKEKR AQRKAAKAAK KAAQSKQ* Computer analysis of this amino acid sequence predicts several transmembrane domains, and also gave the following results: Homology with the dedA homologue of H.influenzae (accession number P452801 ORF78 and the dedA homologue show 58% aa identity in 144aa overlap: Or 78: 4 FLEAFFVEYGYAAVFFVLVICGFGVPIPEDLTLVTGGVISGM—GYTNPHIMFAVGMLGV 61 FL FF EYGY AV FVL+ICGFGVPIPED+TLV+GGVI+G+ N H+M V M+GV DedA: 20 FLIGFFTEYGYWAVLFVLIICGFGVPIPEDITLVSGGVIAGLYPENVNSHLMLLVSMIGV 79 Orf78: 62 LVGDGIMFAAGRIWGQXXLXFXPIAXIMTPXRYEQVQEKFDKYGNWVLFVARFLPGLRTA 121 L GD M+ GRI+G L F PI I+T R V+EKF +YGN VLFVARFLPGLR DedA: 80 LAGDSCMYWLGRIYGTKILRFRPIRRIVTLQRLRMVREKFSQYGNRVLFVARFLPGLRAP 139 Orf78: 122 VFVTAGISRKVSYLRFIIMDGLAA 145 +++ +GI+R+VSY+RF+++D AA DedA: 140 IYMVSGITRRVSYVRFVLIDFCAA 163 Homology with a predicted ORF from N.meninsitidis (strain A) ORF78 shows 93.8% identity over a 145aa overlap with an ORF (ORF78a) from strain A of N. meningitidis: 10 20 30 40 50 60 orf 78 . pep MFAFLEAFFVEYGYAAVFFVLVICGFGVPIPEDLTLVTGGVISGMGYTΝPHIMFAVGMLG I I I : I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I orf78a MFALLEAFFVEYGYAAVFFVLVICGFGVPIPEDLTLVTGGVISGMGYTΝPHIMFAVGMLG 10 20 30 40 50 60 70 80 90 100 110 120 orf 78 . pep VLVGDGIMFAAGRIWGQXXLXFXPIAXIMTPXRYEQVQEKFDKYGΝWVLFVARFLPGLRT II I I I II I II II I II I I I I III I II I II I I I I I I I I I I II I I I I II II I I I I I orf78a VLVGDGIMFAAGRIWGQKILKFKPIARIMTPKRYAQVQEKFDKYGΝWVLFVARFLPGLRT 70 30 100 110 120 130 140 orf78 . pep AVFVTAGISRKVSYLRFIIMDGLAA I I II I I II I I I I II I IM II I I I II orf78a AVFVTAGISRKVSYLRFLIMDGLAALISVPVWIYLGEYGAHΝIDWLMAKMHSLQSGIFIA 130 140 150 160 170 180 The complete length ORF78a nucleotide sequence <SEQ ID 723> is: 1 ATGTTTGCCC TTTTGGAAGC CTTTTTTGTC GAATACGGCT ATGCGGCCGT 51 GTTTTTCGTT TTGGTCATCT GCGGTTTCGG CGTGCCGATT CCCGAGGATT 101 TGACCTTGGT AACAGGCGGC GTGATTTCGG GTATGGGTTA TACCAATCCG 151 CATATTATGT TTGCAGTCGG TATGCTCGGC GTATTGGTCG GGGACGGCAT 201 CATGTTCGCC GCCGGACGCA TCTGGGGGCA GAAAATCCTC AAGTTCAAAC 251 CGATTGCGCG CATCATGACG CCGAAACGTT ACGCACAGGT TCAGGAAAAA 301 TTCGACAAAT ACGGCAACTG GGTGTTATTT GTCGCTCGTT TCCTGCCCGG 351 TTTGCGGACT GCCGTTTTCG TTACCGCCGG CATCAGCCGC AAAGTATCGT 401 ATCTGCGCTT TCTGATTATG GACGGGCTTG CCGCGCTGAT TTCCGTGCCC 451 GTTTGGATTT ACTTGGGCGA GTACGGCGCG CACAACATCG ATTGGCTGAT 501 GGCGAAAATG CACAGCCTGC AATCCGGCAT CTTCATCGCA TTGGGCGTGC 551 TGGCGGCGGC GCTGGCGTGG TTCTGGTGGC GCAAACGCCG ACATTATCAG 601 CTTTACCGCG CACAATTGAG CGAAAAACGC GCCAAACGCA AGGCGGAAAA 651 GGCAGCGAAA AAAGCGGCAC AGAAGCAGCA GTAA This encodes a protein having amino acid sequence <SEQ ID 724>: 1 MFALLEAFFV EYGYAAVFFV LVICGFGVPI PEDLTLVTGG VISGMGYTNP 51 HIMFAVGMLG VLVGDGIMFA AGRIWGQKIL KFKPIARIMT PKRYAQVQEK 101 FDKYGNWVLF VARFLPGLRT AVFVTAGISR KVSYLRFLIM DGLAALISVP 151 VWIYLGEYGA HNIDWLMAKM HSLQSGIFIA LGVLAAALAW FWWRKRRHYQ 201 LYRAQLΞEKR AKRKAEKAAK KAAQKQQ* ORF78a and ORF78-1 show 89.0% identity in 227 aa overlap: 10 20 30 40 50 60 orf78a . pep MFALLEAFFVEYGYAAVFFVLVICGFGVPIPEDLTLVTGGVISGMGYTNPHIMFAVGMLG II I: II II I I II I I I I II I II I I I I II II I I I I II I I I I II I I I I II I I II I I I I I I I II orf78-1 MFAFLEAFFVEYGYAAVFFVLVICGFGVPIPEDLTLVTGGVISGMGYTNPHIMFAVGMLG 10 20 30 40 50 60 70 80 90 100 110 120 orf78a. pep VLVGDGIMFAAGRIWGQKILKFKPIARIMTPKRYAQVQEKFDKYGNWVLFVARFLPGLRT I I I I I I I I I II I I II I I I II : I II II I II II I I I I I I I II I II I II I II I I I I I I I I II orf78-1 VLVGDGIMFAAGRIWGQKILRFKPIARIMTPKRYEQVQEKFDKYGNWVLFVARFLPGLRT 70 80 90 100 110 120 130 140 150 160 170 180 orf78a. pep AVFVTAGISRKVSYLRFLIMDGLAALISVPVWIYLGEYGAHNIDWLMAKMHSLQSGIFIA II I I II I I II I I II I I I : I I II I II I I I II : I II I I I II I I I I I I II I II I I I I I I I I : orf78-1 AVFVTAGISRKVSYLRFIIMDGLAALISVPIWIYLGEYGAHNIDWLMAKMHSLQSGIFVI 130 140 150 160 170 180 190 200 210 220 orf78a . pep LGVLAAALAWFWWRKRRHYQLYRAQLSEKRAKRKAEKAAKKAAQKQQX II: I::: I I : I I: IM: I : I I : : I : I I I I : II I I I I I I I I I :: II orf78-1 LGIGATWAWIWWKKRQRIQFYRSKLKEKRAQRKAAKAAKKAAQSKQX 190 200 210 220 Homology with a predicted ORF from N. onorrhoeae ORF78 shows 97.4% identity over 38 aa overlap with a predicted ORF (ORF78ng) from N. gonorrhoeae: orf78.pep XXLXFXPIAXIMTPXRYEQVQEKFDKYGNWVLFVARFLPGLRTAVFVTAGISRKVSYLRF 137 I II I I I I I I I II I I I I I I I I II I II II II I orf78ng YPVLFVARFLPGLRTAVFVTAGISRKVSYLRF 32 orf78.pep IIMDGLAA 145 : II I I II I orf78ng LIMDGLAALISVPVWIYLGEYGAHNIDWLMAKMHSLQSGIFIALGVLAAALAWFWWRKRR 92 The ORF78ng nucleotide sequence <SEQ ID 725> is predicted to encode a protein comprising amino acid sequence <SEQ ID 726>: 1 .YPVLFVARFL PGLRTAVFVT AGISRKVSYL RFLIMDGLAA LISVPVWIYL 51 GEYGAHNIDW LMAKMHSLQS GIFIALGVLA AALAWFWWRK RRHYQLYRAQ 101 LSEKRAKRKA EKAAKKAAQK QQ* Further work revealed the complete gonococcal nucleotide sequence <SEQ ID 727>: 1 atgtttgccc tttTggaagc CTTTTTTGTC GAAtacggCt atgcGGCCGT 51 GTTTTTCGTT TTGGTCATCT GCGGTTTCGG CGTGCCGATT CCCGAAGATT 101 TGACCTTGGT AACGGGCGGC GTGATTTCGG GTATGGGTTA TACCAATCCG 151 CATATTATGT TTGCGGTCGG TATGCTCGGC GTGTTGGCGG GCGACGGCGT 201 GATGTTTGCC GCCGGACGCA TCTGGGGGCA GAAAATCCTC AAGTTCAAAC 251 CGATTGCGCG CATCATGACG CCGAAACGTT ACGCGCAGGT TCAGGAAAAA 301 TTCGACAAAT ACGGCAACTG GGTTCTGTTT GTCGCCCGTT TCCTGCCGGG 351 TTTGCGGACT GCCGTTTTCG TTACCGCCGG CATCAGCCGC AAAGTATCGT 401 ATCTGCGCTT TCTGATTATG GACGGGCTGG CCGCGCTGAT TTCCGTGCCC 451 GTTTGGATTT ACTTGGGCGA GTACGGCGCG CACAACATCG ATTGGCTGAT 501 GGCGAAAATG CACAGCCTGC AATCGGGCAT CTTCATCGCA TTGGGCGTGC 551 TGGCGGCGGC GCTGGCGTGG TTCTGGTGGC GCAAACGCCG ACATTATCAG 601 CTTTACCGCG CACAATTGAG CGAAAAACGC GCCAAACGCA AGGCGGAAAA 651 GGCAGCGAAA AAAGCGGCAC AGAAGCAGCA GTAa This coπesponds to the amino acid sequence <SEQ LD 728; ORF78ng-l>: 1 MFALLEAFFV EYGYAAVFFV LVICGFGVPI PEDLTLVTGG VISGMGYTNP 51 HIMFAVGMLG VLAGDGVMFA AGRIWGQKIL KFKPIARIMT PKRYAQVQEK 101 FDKYGNWVLF VARFLPGLRT AVFVTAGISR KVSYLRFLIM DGLAALISVP 151 VWIYLGEYGA HNIDWLMAKM HSLQSGIFIA LGVLAAALAW FWWRKRRHYQ 201 LYRAQLSEKR AKRKAEKAAK KAAQKQQ* ORF78ng-l and ORF78-1 show 88.1% identity in 227 aa overlap: 10 20 30 40 50 60 orf78-1. pep MFAFLEAFFVEYGYAAVFFVLVICGFGVPIPEDLTLVTGGVISGMGYTNPHIMFAVGMLG I II: II II II II I II II I I I II I I II I I I II II I I II I II II II II II I I I II II II II I orf78ng-l MFALLEAFFVEYGYAAVFFVLVICGFGVPIPEDLTLVTGGVISGMGYTNPHIMFAVGMLG 10 20 30 40 50 60 70 80 90 100 110 120 orf78-1. pep VLVGDGIMFAAGRIWGQKILRFKPIARIMTPKRYEQVQEKFDKYGNWVLFVARFLPGLRT I I: I I M I I I I I I I II I I I I M I II I II I I I II I I I I II II I I II II I I I II II II II I orf78ng-l VLAGDGVMFAAGRIWGQKILKFKPIARIMTPKRYAQVQEKFDKYGNWVLFVARFLPGLRT 70 80 90 100 110 120 130 140 150 160 170 180 orf78-1. pep AVFVTAGISRKVSYLRFIIMDGLAALISVPIWIYLGEYGAHNIDWLMAKMHSLQSGIFVI I II I II II I I II I I I I I : I I I II I I I II I I : I I I I I II I I I II I I II I II I I I II I I I : orf78ng-l AVFVTAGISRKVSYLRFLIMDGLAALISVPVWIYLGEYGAHNIDWLMAKMHSLQSGIFIA 130 140 150 160 170 180 190 200 210 220 orf78-1. pep LGIGATVVAWIWWKKRQRIQFYRSKLKEKRAQRKAAKAAKKAAQSKQX II: I::: I I : I I : I I :: I: IM: I : I II I : I II I I I II II I :: I I orf78ng-1 LGVLAAALAWFWWRKRRHYQLYRAQLSEKRAKRKAEKAAKKAAQKQQX 190 200 210 220 Furthermore, orf78ng-l shows homology to the dedA protein from H.influenzae: sp|P45280|YG29_HAEIN HYPOTHETICAL PROTEIN HI1629 >gi | 1073983 | pir || D64133 dedA protein (dedA) homolog - Haemophilus influenzae (strain Rd KW20) >gi 11574476 (U32836) dedA protein (dedA) [Haemophilus influenzae] Length = 212 Score = 223 bits (563), Expect = 7e-58 Identities = 108/182 (59%), Positives = 140/182 (76%), Gaps = 2/182 (1%) Query: 5 LEAFFVEYGYAAVFFVLVICGFGVPIPEDLTLVTGGVISGM—GYTNPHIMFAVGMLGVL 62 L FF EYGY AV FVL+ICGFGVPIPED+TLV+GGVI+G+ N H+M V M+GVL Sbjct 21 LIGFFTEYGYWAVLFVLIICGFGVPIPEDITLVSGGVIAGLYPENVNSHLMLLVSMIGVL 80 Query: 63 AGDGVMFAAGRIWGQKILKFKPIARIMTPKRYAQVQEKFDKYGNWVLFVARFLPGLRTAV 122 AGD M+ GRI+G KIL+F+PI RI+T +R V+EKF +YGN VLFVARFLPGLR + Sbjct: 81 AGDSCMYWLGRIYGTKILRFRPIRRIVTLQRLRMVREKFSQYGNRVLFVARFLPGLRAPI 140 Query: 123 FVTAGISRKVSYLRFLIMDGLAALISVPVWIYLGEYGAHNIDWLMAKMHSLQSGIFIALG 182 ++ +GI+R+VSY+RF+++D AA+ISVP+WIYLGE GA N+DWL ++ Q I+I +G Sbjct: 141 YMVSGITRRVSYVRFVLIDFCAAIISVPIWIYLGELGAKNLDWLHTQIQKGQIVIYIFIG 200 Query: 183 VL 184 L Sbjct: 201 YL 202 Based on this analysis, including the presence of putative transmembrane domains, it is predicted that these proteins from N. meningitidis and N.gonorrhoeae, and their epitopes, could be useful antigens for vaccines or diagnostics, or for raising antibodies. Example 87 The following partial DNA sequence was identified in N. meningitidis <SEQ ID 729>: 1 ATGAAAAAAT TATTGGCGGC CGTGATGATG GCAGGTTTGG CAGGCGCGGT 51 TTCCGCCGCC GGAGTCCACG TTGAGGACGG CTGGGCGCGC ACCACCGTCG 101 AAGGTATGAA AATAGGCGGC GCGTTCATGA AAATCCACAA CGACGAAGCC 151 AAACAAGACT TTTTGCTCGG CGGAAGCAGC CCCGTTGCCG ACCGCGTCGA 201 AGTGCATACC CACATCAACG ACAACGGCGT GATGCGGATG CGCGAAGTCG 251 AAGGCGGCGT GCCTTTGGAA GCGAAATCCG TTACCGAACT CAAACCCGGC 301 AGCTATCATG TGATGTTTAT GGGTTTGAAA AAACAATTAA AAGAGGGCGA 351 TAAAATTCCC GTTACCCTGA AATTTAAAAA CGCCAAAGCG CAAACCGTCC 401 AACTGGAAGT CAAAATCGCG CCGATGCCGG CAATGAACCA C... This coπesponds to the amino acid sequence <SEQ ED 730; ORF79>: 1 MKKLLAAVMM AGLAGAVSAA GVHVEDGWAR TTVEGMKIGG AFMKIHNDEA 51 KQDFLLGGSS PVADRVEVHT HINDNGVMRM REVEGGVPLE AKSVTELKPG 101 SYHVMFMGLK KQLKEGDKIP VTLKFKNAKA QTVQLEVKIA PMPAMNH.. Further work revealed the complete nucleotide sequence <SEQ ID 731>: 1 ATGAAAAAAT TATTGGCGGC CGTGATGATG GCAGGTTTGG CAGGCGCGGT 51 TTCCGCCGCC GGAGTCCACG TTGAGGACGG CTGGGCGCGC ACCACCGTCG 101 AAGGTATGAA AATAGGCGGC GCGTTCATGA AAATCCACAA CGACGAAGCC 151 AAACAAGACT TTTTGCTCGG CGGAAGCAGC CCCGTTGCCG ACCGCGTCGA 201 AGTGCATACC CACATCAACG ACAACGGCGT GATGCGGATG CGCGAAGTCG 251 AAGGCGGCGT GCCTTTGGAA GCGAAATCCG TTACCGAACT CAAACCCGGC 301 AGCTATCATG TGATGTTTAT GGGTTTGAAA AAACAATTAA AAGAGGGCGA 351 TAAAATTCCC GTTACCCTGA AATTTAAAAA CGCCAAAGCG CAAACCGTCC 401 AACTGGAAGT CAAAATCGCG CCGATGCCGG CAATGAACCA CGGTCATCAC 451 CACGGCGAAG CGCATCAGCA CTAA This coπesponds to the amino acid sequence <SEQ ID 732; ORF79-l>: 1 MKKLLAAVMM AGLAGAVSAA GVHVEDGWAR TTVEGMKIGG AFMKIHNDEA 51 KQDFLLGGSS PVADRVEVHT HINDNGVMRM REVEGGVPLE AKSVTELKPG 101 SYHVMFMGLK KQLKEGDKIP VTLKFKNAKA QTVQLEVKIA PMPAMNHGHH 151 HGEAHQH* Computer analysis of this amino acid sequence revealed a putative leader peptide and also gave the following results: Homology with a predicted ORF from N.meninsitidis (strain A) ORF79 shows 94.6% identity over a 147aa overlap with an ORF (ORF79a) from strain A of N. meningitidis: 10 20 30 40 50 60 orf79. pep MKKLLAAVMMAGLAGAVSAAGVHVEDGWARTTVEGMKIGGAFMKIHNDEAKQDFLLGGSS II I I I I I II I I II I I II II I: II I I I I II II I II I I : II I I I II II II I II II I I I I I I orf79a MKXLLAAVMMAGLAGAVSAAGIHVEDGWARTTVEGMKMGGAFMKIHNDEAKQDFLLGGSS 10 20 30 40 50 60 70 80 90 100 110 120 orf79. pep PVADRVEVHTHINDNGVMRMREVEGGVPLEAKSVTELKPGSYHVMFMGLKKQLKEGDKIP II I I I I I II I II II I I I I I I I I I II I I II II II I II I II II I II II I I II I I I I I II I orf79a PVADRVEVHTHINDNGVMRMREVEGGVPLEAKSVTELKPGSYHVMFMGXKKQLKXGDKIP 70 80 90 100 110 120 130 140 orf79. pep VTLKFKNAKAQTVQLEVKIAPMPAMNH II II II I I II II I II II I III MM orf79a VTLKFKNAKAQTVQLEVKTAPMSAMDHGHHHGEAHQHX 130 140 150 The complete length ORF79a nucleotide sequence <SEQ LD 733> is: 1 ATGAAANAAC TATTGGCAGC CGTGATGATG GCAGGTTTGG CAGGCGCGGT 51 TTCCGCCGCC GGAATCCACG TTGAGGACGG CTGGGCGCGC ACCACCGTCG 101 AAGGTATGAA AATGGGCGGC GCGTTCATGA AAATCCACAA CGACGAAGCC 151 AAACAAGACT TTTTGCTCGG CGGAAGCAGC CCTGTTGCCG ACCGCGTCGA 201 AGTGCATACC CATATCAATG ATAACGGTGT GATGCGGATG CGCGAAGTCG 251 AAGGCGGCGT GCCTTTGGAG GCGAAATCCG TTACCGAACT CAAACCCGGC 301 AGCTATCATG TCATGTTTAT GGGTNTGAAA AAACAATTAA AAGANGGCGA 351 CAAGATTCCC GTTACCCTGA AATTTAAAAA CGCCAAAGCA CAAACCGTCC 401 AACTGGAAGT CAAAACCGCG CCGATGTCGG CAATGGACCA CGGTCATCAC 451 CACGGCGAAG CGCATCAGCA CTAA This encodes a protein having amino acid sequence <SEQ LD 734>: 1 MKXLLAAVMM AGLAGAVSAA GIHVEDGWAR TTVEGMKMGG AFMKIHNDEA 51 KQDFLLGGSS PVADRVEVHT HINDNGVMRM REVEGGVPLE AKSVTELKPG 101 SYHVMFMGXK KQLKXGDKIP VTLKFKNAKA QTVQLEVKTA PMSAMDHGHH 151 HGEAHQH* ORF79a and ORF79-1 show 94.9% identity in 157 aa overlap: 10 20 30 40 50 60 orf79a . pep MKXLLAAVMMAGLAGAVSAAGIHVEDGWARTTVEGMKMGGAFMKIHNDEAKQDFLLGGSS II I I I I II I I I II I I II I II : I I I II I II I I I II I I : I II I II I I I I II I I II I II I I I orf79-1 MKKLLAAVMMAGLAGAVSAAGVHVEDGWARTTVEGMKIGGAFMKIHNDEAKQDFLLGGSS 10 20 30 40 50 60 70 80 90 100 110 120 orf79a . pep PVADRVEVHTHINDNGVMRMREVEGGVPLEAKSVTELKPGSYHVMFMGXKKQLKXGDKIP orf79-1 PVADRVEVHTHINDNGVMRMREVEGGVPLEAKSVTELKPGSYHVMFMGLKKQLKEGDKIP 70 80 90 100 110 120 130 140 150 orf79a .pep VTLKFKNAKAQTVQLEVKTAPMSAMDHGHHHGEAHQHX I II I I II I II I I II I II I III I I : I I II I I I II I II orf79-1 VTLKFKNAKAQTVQLEVKIAPMPAMNHGHHHGEAHQHX 130 140 150 Homology with a predicted ORF from N.sonorrhoeae ORF79 shows 96.1% identity over 76 aa overlap with a predicted ORF (ORF79ng) from N.gonorrhoeae: orf 9. pep FMKIHNDEAKQDFLLGGSSPVADRVEVHTHINDNGVMRMREVEGGVPLEAKSVTELKPGS 101 I II I I I I I I I I I : I I I II I I II I I II I II I orf79ng INDNGVMRMREVKGGVPLEAKSVTELKPGS 30 orf79.pep YHVMFMGLKKQLKEGDKIPVTLKFKNAKAQTVQLEVKIAPMPAMNH 147 I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I orf79ng YHVMFMGLKKQLKEGDKIPVTLKFKNAKAQTVQLEVKTAPMSAMNHGHHHGEAHQH 86 An ORF79ng nucleotide sequence <SEQ ID 735> was predicted to encode a protein comprising amino acid sequence <SEQ ID 736>: 1 ..INDNGVMRMR EVKGGVPLEA KSVTELKPGS YHVMFMGLKK QLKEGDKIPV 51 TLKFKNAKAQ TVQLEVKTAP MSAMNHGHHH GEAHQH* Further work revealed the complete gonococcal DNA sequence <SEQ LD 737>: 1 ATGAAAAAAT TATTGGCAGC CGTGATGATG GCAGGTTTGG CAGGCGCGGT 51 TTccgccgCc GGagTccAtG TCGAggACGG CTGGGCGCGC accaCTGtcg 101 aaggtATgaa aatggGCGGC GCgttCATga aaATCCACAA CGACGaaGcc 151 atacaaGACt ttgtgcTCgg CGGaagcatg cccgttgccg accgcGTCGA 201 AGTGCAtaca cacATCAACG ACAACGGCGT GATGCGTATG CGCGAAGTCA 251 AAGGCGGCGT GCCTTTGGAG GCGAAATCCG TTACCGAACT CAAACCCGGC 301 AGCTATCACG TGATGTTTAT GGGTTTGAAA AAACAACTGA AAGAGGGCGA 351 CAAGATTCCC GTTACCCTGA AATTTAAAAA CGCCAAAGCG CAAACCGTCC 401 AACTGGAAGT CAAAACCGCG CCGATGTCGG CAATGAACCA CGGTCATCAC 451 CACGGCGAAG CGCATCAGCA CTAA This coπesponds to the amino acid sequence <SEQ ID 738; ORF79ng-l>: 1 MKKLLAAVMM AGLAGAVSAA GVHVEDGWAR TTVEGMKMGG AFMKIHNDEA 51 IQDFVLGGSM PVADRVEVHT HINDNGVMRM REVKGGVPLE AKSVTELKPG 101 SYHVMFMGLK KQLKEGDKIP VTLKFKNAKA QTVQLEVKTA PMSAMNHGHH 151 HGEAHQH* ORF79ng-l and ORF79-1 show 95.5% identity in 157 aa overlap: 10 20 30 40 50 60 orf79-1. pep MKKLLAAVMMAGLAGAVSAAGVHVEDGWARTTVEGMKIGGAFMKIHNDEAKQDFLLGGSS I II I II I I I II II II II I I I I II I II I I I I I II I I II : I II II I I I I I I I I I I: II II orf79ng-1 MKKLLAAVMMAGLAGAVSAAGVHVEDGWARTTVEGMKMGGAFMKIHNDEAIQDFVLGGSM 10 20 30 40 50 60 70 80 90 100 110 120 orf79-1. pep PVADRVEVHTHINDNGVMRMREVEGGVPLEAKSVTELKPGSYHVMFMGLKKQLKEGDKIP II II I I I I I I II II II I I I II I I : I I I I I I II I II I II I I I I II II I II II I II I I I I I I orf79ng-l PVADRVEVHTHINDNGVMRMREVKGGVPLEAKSVTELKPGSYHVMFMGLKKQLKEGDKIP 70 80 90 100 110 120 130 140 150 orf79-1. pep VTLKFKNAKAQTVQLEVKIAPMPAMNHGHHHGEAHQHX II I II I I I I I I I I II II I III I I II II I I I II I I I I orf79ng-l VTLKFKNAKAQTVQLEVKTAPMSAMNHGHHHGEAHQHX 130 140 150 Furthermore, ORF79ng-l shows significant homology to a protein from Aquifex aeolicus: gi 12983695 (AE000731) putative protein [Aquifex aeolicus] Length = 151 Score = 63.6 bits (152), Expect = 6e-10 Identities = 38/114 (33%), Positives = 58/114 (50%), Gaps = 1/114 (0%) Query: 24 VEDGWARTTVEGMKMGGAFMKIHNDEAIQDFVLGGSMPVADRVEVHTHINDNGVMRMREV 83 V+ W G M I N+ D+++G +A RVE+H + +N V +M Sbjct: 27 VKHPWVMEPPPGPNTTMMGMIIVNEGDEPDYLIGAKTDIAQRVELHKTVIENDVAKMVPQ 86 Query: 84 KGGVPLEAKSVTELKPGSYHVMFMGLKKQLKEGDKIPVTLKFKNAKAQTVQLEV 137 + + + K E K YHVM +GLKK++KEGDK+ V L F+ + TV+ V Sbjct: 87 ER-IEIPPKGKVEFKHHGYHVMIIGLKKRIKEGDKVKVELIFEKSGKITVEAPV 139 Based on this analysis, it is predicted that the proteins from N. meningitidis and N.gonorrhoeae, and their epitopes, could be useful antigens for vaccines or diagnostics, or for raising antibodies. ORF79-1 (15.6kDa) was cloned in the pET vector .and expressed in E.coli, as described above. The products of protein expression and purification were analyzed by SDS-PAGE. Figure 18 A shows the results of affinity purification of the His-fusion protein. Purified His-fusion protein was used to immunise mice, whose sera were used for ELISA (positive result) and FACS analysis (Figure 18B) These experiments confirm that ORF79-1 is a surface-exposed protein, and that it is a useful immunogen. Example 88 The following DNA sequence, believed to be complete, was identified in N. meningitidis <SEQ ED 739>: 1 ATGACGGTAA CTGCGGCCGA AGGCGGCAAA GCTGCCAAGG CGTTAAAAAA 51 ATATCTGATT ACGGGCATTT TGGTCTGGCT GCCGATTGCG GTAACGGTTT 101 GGGTGGTTTC CTATATCGTT TCCGCGTCCG ATCAGCTCGT CAACCTGCTG 151 CCGAAGCAAT GGCGGCCGCA ATATGTTTTG GGGTTTAATA TCCCGGGGCT 201 GGGCGTTATC GTTGCCATTG CCGTATTGTT TGTAACCGGA TTGTTTGCCG 251 CCAACGTATT GGGTCGGCAG ATCCTCGCCG CGTGGGACAG CCTGTTGGGG 301 CGGATTCCGG TTGTGAAAtC CATCTATTCG AGTGTGAAAA AAGTATCCGA 351 ATacgTGCTG TCCGACAGCA GCCGTTCGTT TAAAACGCCG GTACTCGTGC 401 CGTTTCCCCA GCCCGGTATT TGGACGATyG CTTTCGTGTC AGGGCAGGTG 451 TCGAATGCGG TTAAGGCCGC ATTGCCGAAs GACGGCGATT ATCTTTCCGT 501 GTATGTTCCG ACCACGCCGA ATCCGACCGG CGGTTACTAT ATTATGGTAA 551 AGAAAAGCGA TGTGCGCGAA CTCGATATGA GCGTGGACGA AsCATTGAAA 601 TATGTGATTT CGCTGGGTAT GGTCATCCCT GACGACCTGC CCGTCAAAAC 651 ATTGGCAsGA CCTATGCCGT CTGAAAAGGC GGATTTGCCC GAACAACAAT 701 AA This coπesponds to the amino acid sequence <SEQ ID 740; ORF98>: 1 MTVTAAEGGK AAKALKKYLI TGILVWLPIA VTVWWSYIV SASDQLVNLL 51 PKQWRPQYVL GFNIPGLGVI VAIAVLFVTG LFAANVLGRQ ILAAWDSLLG 101 RIPWKSIYS SVKKVSEYVL SDSSRSFKTP VLVPFPQPGI WTIAFVSGQV 151 SNAVKAALPX DGDYLSVYVP TTPNPTGGYY IMVKKSDVRE LDMSVDEXLK 201 YVISLGMVIP DDLPVKTLAX PMPSEKADLP EQQ* Further work revealed the complete nucleotide sequence <SEQ ID 741>: 1 ATGACGGAAC nTGCGGCCGA AGGCGGCAAA GCTGCCAArG CGTTAAAAAA 51 ATATCTGATT ACGGGCATTT TGGTCTGGCT GCCGATTGCG GTAACGGTTT 101 GGGTGGTTTC CTATATCGTT TCCGCGTCCG ATCAGCTCGT CAACCTGCTG 151 CCGAAGCAAT GGCGGCCGCA ATATGTTTTG GGGTTTAATA TCCCGGGGCT 201 GGGCGTTATC GTTGCCATTG CCGTATTGTT TGTAACCGGA TTGTTTGCCG 251 CCAACGTATT GGGTCGGCAG ATCCTCGCCG CGTGGGACAG CCTGTTGGGG 301 CGGATTCCGG TTGTGAAATC CATCTATTCG AGTGTGAAAA AAGTATCCGA 351 ATCGCTGCTG TCCGACAGCA GCCGTTCGTT TAAAACGCCG GTACTCGTGC 401 CGTTTCCCCA GCCCGGTATT TGGACGATTG CTTTCGTGTC AGGGCAGGTG 451 TCGAATGCGG TTAAGGCCGC ATTGCCGAAG GACGGCGATT ATCTTTCCGT 501 GTATGTTCCG ACCACGCCGA ATCCGACCGG CGGTTACTAT ATTATGGTAA 551 AGAAAAGCGA TGTGCGCGAA CTCGATATGA GCGTGGACGA AGCATTGAAA 601 TATGTGATTT CGCTGGGTAT GGTCATCCCT GACGACCTGC CCGTCAAAAC 651 ATTGGCAGGA CCTATGCCGT CTGAAAAGGC GGATTTGCCC GAACAACAAT 701 AA This coπesponds to the .amino acid sequence <SEQ ID 742; ORF98-l>: 1 MTEXAAEGGK AAKALKKYLI TGILVWLPIA VTVWWSYIV SASDQLVNLL 51 PKQWRPQYVL GFNIPGLGVI VAIAVLFVTG LFAANVLGRQ ILAAWDSLLG 101 RIPWKSIYS SVKKVSESLL SDSSRSFKTP VLVPFPQPGI WTIAFVSGQV 151 SNAVKAALPK DGDYLSVYVP TTPNPTGGYY IMVKKSDVRE LDMSVDEALK 201 YVISLGMVIP DDLPVKTLAG PMPSEKADLP EQQ* Computer analysis of this amino acid sequence gave the following results: Homology with a predicted ORF from N.meninsitidis (sfrain Al ORF98 shows 96.1% identity over a 233aa overlap with an ORF (ORF98a) from strain A of N. meningitidis: 10 20 30 40 50 60 orf98.pep MTVTAAEGGKAAKALKKYLITGILVWLPIAVTVWWSYIVSASDQLVNLLPKQWRPQYVL M II II I I I I I II II I II I I II I II I II II I II I I I I I I I I I I II II I I II II I I I I I orf98a MTEPAAEGGKAAKALKKYLITGILVWLPIAVTVWWSYIVSASDQLVNLLPKQWRPQYVL 10 20 30 40 50 60 70 80 90 100 110 120 orf98. pep GFNIPGLGVIVAIAVLFVTGLFAANVLGRQILAAWDSLLGRIPWKSIYSSVKKVSEYVL I II I II II II I I I I I I I II I II I II I II I I I II II I I II I II I I I I I I II I I I I II : I orf98a GFNIPGLGVIVAIAVLFVTGLFAANVLGRQILAAWDSLLGRIPWKSIYSSVKKVSXSLL 70 80 90 100 110 120 130 140 150 160 170 180 orf98. pep SDSSRSFKTPVLVPFPQPGIWTIAFVSGQVSNAVKAALPXDGDYLSVYVPTTPNPTGGYY I II I I I II I I I I I II I I II I II I II I II I II II II I I I I II II II I I I II I II II II I orf98a SDSSRSFKTPVLVPFPQSGIWTIAFVSGQVSNAVKAALPKDGDYLSVYVPTTPNPTGGYY 130 140 150 160 170 180 190 200 210 220 230 orf98.pep IMVKKSDVRELDMSVDEXLKYVISLGMVIPDDLPVKTLAXPMPSEKADLPEQQX I I II I II I I II I I I II I I II I I I I II I II I I I II I I II I I I I II I I I I I II I orf98a IMVKKSDVRELDMSVDEALKYVISLGMVIPDDLPVKTLAGPMPSEKADLPEQQX 190 200 210 220 230 The complete length ORF98a nucleotide sequence <SEQ ID 743> is: 1 ATGACGGAAC CTGCGGCCGA AGGCGGCAAA GCTGCCAAGG CGTTAAAAAA 51 ATATCTGATT ACGGGCATTT TGGTCTGGCT GCCGATTGCG GTAACGGTTT 101 GGGTGGTTTC CTATATCGTT TCCGCGTCCG ATCAGCTCGT CAACCTGCTG 151 CCGAAGCAAT GGCGGCCGCA ATATGTTTTG GGGTTTAATA TCCCGGGGCT 201 GGGCGTTATC GTTGCCATTG CCGTATTGTT TGTAACCGGA TTATTTGCCG 251 CAAACGTATT GGGCCGGCAG ATTCTTGCCG CGTGGGACAG CTTGTTGGGG 301 CGGATTCCGG TTGTGAAGTC CATCTATTCG AGTGTGAAAA AAGTATCCGA 351 NTCGTTGCTG TCCGACAGCA GCCGTTCGTT TAAAACACCA GTACTCGTGC 401 CGTTTCCCCA ATCGGGTATT TGGACAATCG CATTCGTGTC CGGTCAGGTG 451 TCGAATGCGG TTAAGGCCGC ATTGCCGAAG GACGGCGATT ATCTTTCCGT 501 GTATGTTCCG ACCACGCCGA ATCCGACCGG CGGTTACTAT ATTATGGTAA 551 AGAAAAGCGA TGTGCGCGAA CTCGATATGA GCGTGGACGA AGCGTTGAAA 601 TATGTGATTT CGCTGGGTAT GGTCATCCCT GACGACCTGC CCGTCAAAAC 651 ATTGGCAGGA CCTATGCCGT CTGAAAAGGC GGATTTGCCC GAACAACAAT 701 AA This encodes a protein having amino acid sequence <SEQ LD 744>: 1 MTEPAAEGGK AAKALKKYLI TGILVWLPIA VTVWWSYIV SASDQLVNLL 51 PKQWRPQYVL GFNIPGLGVI VAIAVLFVTG LFAANVLGRQ ILAAWDSLLG 101 RIPWKSIYS SVKKVSXSLL SDSSRSFKTP VLVPFPQSGI WTIAFVSGQV 151 SNAVKAALPK DGDYLSVYVP TTPNPTGGYY IMVKKSDVRE LDMSVDEALK 201 YVISLGMVIP DDLPVKTLAG PMPSEKADLP EQQ* ORF98a and ORF98-1 show 98.7% identity in 233 aa overlap: 10 20 30 40 50 60 orf98a. pep MTEPAAEGGKAAKALKKYLITGILVWLPIAVTVWWSYIVSASDQLVNLLPKQWRPQYVL III I I I II I I I II I II I II I I II II I I II I II I II II II I I I I I I I I I I I I II I I I I I I orf98-l MTEXAAEGGKAAKALKKYLITGILVWLPIAVTVWWSYIVSASDQLVNLLPKQWRPQYVL 10 20 30 40 50 60 70 80 90 100 110 120 orf98a. pep GFNIPGLGVIVAIAVLFVTGLFAANVLGRQILAAWDSLLGRIPWKSIYSSVKKVSXSLL II I II I II II II I I II I II I II I II I I I II II I II I I I I II II II I I I II I I II I I III orf98-1 GFNIPGLGVIVAIAVLFVTGLFAANVLGRQILAAWDSLLGRIPWKSIYSSVKKVSESLL 70 80 90 100 110 120 130 140 150 160 170 180 orf98a . pep SDSSRSFKTPVLVPFPQSGIWTIAFVSGQVSNAVKAALPKDGDYLSVYVPTTPNPTGGYY I I II I II I I I I I I II I I I II I I I I I I II I II I I I I I I I II I I II I I I II I I I I I I I I I I orf98-1 SDSSRSFKTPVLVPFPQPGIWTIAFVSGQVSNAVKAALPKDGDYLSVYVPTTPNPTGGYY 130 140 150 160 170 180 190 200 210 220 230 orf98a. pep IMVKKSDVRELDMSVDEALKYVISLGMVIPDDLPVKTLAGPMPSEKADLPEQQX II I II I II II II I I I I I II I II I I II I I II I II I I I I I I I I I II I I I I I II I I I orf98-1 IMVKKSDVRELDMSVDEALKYVISLGMVIPDDLPVKTLAGPMPSEKADLPEQQX 190 200 210 220 230 Homology with a predicted ORF from N.sonorrhoeae ORF98 shows 95.3% identity over a 233 aa overlap with a predicted ORF (ORF98ng) from N.gonorrhoeae: 10 20 30 40 50 60 orf98. pep MTVTAAEGGKAAKALKKYLITGILVWLPIAVTVWWSYIVSASDQLVNLLPKQWRPQYVL 60 II I I I I I I I I II I I I I II II I II II I I I II I I II II II I II I I I I I I I I II I I I I I I I orf98ng MTEPAAEGGKAAKALKKYLITGILVWLPIAVTVWWSYIVSASDQLVNLLPKQWRPQYVL 60 orf98.pep GFNIPGLGVIVAIAVLFVTGLFAANVLGRQILAAWDSLLGRIPWKSIYSSVKKVSEYVL 120 I I II I I I II I I I I I II II I I I I II II I I II I I I II I I I I I I I I I I II I I I I I I I I I : I orf98ng GFNIPGLGVIVAIAVLFVTGLFAANVLGRQILAAWDSLLXRIPWKSIYSSVKKVSESLL 120 orf98.pep SDSSRSFKTPVLVPFPQPGIWTIAFVSGQVSNAVKAALPXDGDYLSVYVPTTPNPTGGYY 180 II I I I II I II II II II I II II I II II II II I I I I I I II I I II I I II I I II I II II I II orf98ng SDSSRSFKTPVLVPFPQSGIWTIAFVSGQVSNAVKAALPQDGDYLSVYVPTTPNPTGGYY 180 orf98. pep IMVKKSDVRELDMSVDEXLKYVISLGMVIPDDLPVKTLAXPMPSEKADLPEQQ 233 I I I I I I I I II I I II I I I I I I II I II II I I I I I II I I I I III I I I : II I II orf98ng IMVKKSDVRELDMSVDEALKYVISLGMVIPDDLPVKTLAGPMPPEKAELPEQQ 233 The complete length ORF98ng nucleotide sequence <SEQ LD 745> is predicted to encode a protein having amino acid sequence <SEQ LD 746>: 1 MTEPAAEGGK AAKALKKYLI TGILVWLPIA VTVWWSYIV SASDQLVNLL 51 PKQWRPQYVL GFNIPGLGVI VAIAVLFVTG LFAANVLGRQ ILAAWDSLLX 101 RIPWKSIYS SVKKVSESLL SDSSRSFKTP VLVPFPQSGI WTIAFVSGQV 151 SNAVKAALPQ DGDYLSVYVP TTPNPTGGYY IMVKKSDVRE LDMSVDEALK 201 YVISLGMVIP DDLPVKTLAG PMPPEKAELP EQQ* Further work revealed the complete nucleotide sequence <SEQ ID 747>: 1 ATGACGGAAC CTGCGGCCGA AGGCGGCAAA GCTGCCAAGG CGTTAAAAAA 51 ATATCTGATT ACAGGCATTT TGGTCTGGCT GCCGATTGCG GTAACGGTTT 101 GGGTGGTTTC CTATATCGTT TCCGCGTCCG ACCAGCTTGT CAACCTGCTG 151 CCGAAGCAAT GGCGGCCGCA ATATGTTTTG GGGTTTAATA TCCCCGGGCT 201 CGGCGTTATT GTTGCCATTG CCGTATTGTT TGTAACCGGA TTATTTGCCG 251 CAAACGTGTT GGGCCGGCAG ATTCTTGCCG CGTGGGACAG CCTGTTgggg 301 cggaTTCCGG TTGTCAAATC CATCTATTCG AGTGTGAAAA AAGTATCCGA 351 ATCGCTGCTG TCCGACAGCA GCCGTTCGTT TAAAACGCCG GTACTCGTGC 401 CGTTTCCCCA ATCGGGTATT TGGACAATCG CATTCGTGTC CGGTCAGGTG 451 TCGAATGCGG TTAAGGCCGC ATTGCCGCAG GATGGCGATT ATCTTTCCGT 501 GTATGTCCCG ACCACGCCCA ACCCGACCGG CGGTTACTAT ATTATGGTAA 551 AGAAAAGCGA TGTGCGCGAA CTCGATATGA GCGTGGACGA AGCGTTGAAA 601 TATGTGATTT CGCTGGGTAT GGTCATCCCT GACGACCTGC CCGTCAAAAC 651 ATTGGCAGGA CCTATGCCGC CTGAAAAGGC GGAGTTGCCC GAACAACAAT 701 AA This coπesponds to the amino acid sequence <SEQ ID 748; ORF98ng-l>: 1 MTEPAAEGGK AAKALKKYLI TGILVWLPIA VTVWWSYIV SASDQLVNLL 51 PKQWRPQYVL GFNIPGLGVI VAIAVLFVTG LFAANVLGRQ ILAAWDSLLG 101 RIPWKSIYS SVKKVSESLL SDSSRSFKTP VLVPFPQSGI WTIAFVSGQV 151 SNAVKAALPQ DGDYLSVYVP TTPNPTGGYY IMVKKSDVRE LDMSVDEALK 201 YVISLGMVIP DDLPVKTLAG PMPPEKAELP EQQ* ORF98ng-l and ORF98-1 show 97.9% identity in 233 aa overlap: 10 20 30 40 50 60 orf98-1.pep MTEXAAEGGKAAKALKKYLITGILVWLPIAVTVWWSYIVSASDQLVNLLPKQWRPQYVL III I I I I II I II I II I I I I I II I II I II II II I I I II I I II I II II I I II I I II I I II I 0rf98ng-l MTEPAAEGGKAAKALKKYLITGILVWLPIAVTVWWSYIVSASDQLVNLLPKQWRPQYVL 10 20 30 40 50 60 70 80 90 100 110 120 orf98-1. pep GFNIPGLGVIVAIAVLFVTGLFAANVLGRQILAAWDSLLGRIPWKSIYSSVKKVSESLL orf98ng-l GFNIPGLGVIVAIAVLFVTGLFAANVLGRQILAAWDSLLGRIPWKSIYSSVKKVSESLL 70 80 90 100 110 120 130 140 150 160 170 180 orf98-1. pep SDSSRSFKTPVLVPFPQPGIWTIAFVSGQVSNAVKAALPKDGDYLSVYVPTTPNPTGGYY I I I II I I I I I I II I II I II I II I II I II II II II I I II : I I I I I I II I I I I I II I I II I orf98ng-l SDSSRSFKTPVLVPFPQSGIWTIAFVSGQVSNAVKAALPQDGDYLSVYVPTTPNPTGGYY 130 140 150 160 170 180 190 200 210 220 230 orf98-1. pep IMVKKSDVRELDMSVDEALKYVISLGMVIPDDLPVKTLAGPMPSEKADLPEQQX II I II I I I II I I II I II II II I I I I II I I I I I II I I II II I I I I I I: I II I II orf98ng-l IMVKKSDVRELDMSVDEALKYVISLGMVIPDDLPVKTLAGPMPPEKAELPEQQX 190 200 210 220 230 Based on this analysis, including the fact that the putative transmembrane domains in the gonococcal protein are identical to the sequences in the memngococcal protein, it is predicted that the proteins from N. meningitidis and N.gonorrhoeae, and their epitopes, could be useful antigens for vaccines or diagnostics, or for raising antibodies. Example 89 The following partial DNA sequence was identified in N. meningitidis <SEQ ID 749>: 1 ATgAAAACGG TAGTCTGGAT TGTCGTCCTG TTTGCCGCCG CCGTCGGACT 51 GGCGCTGGCT TCGGGCATTT ACACCGGCGA CGTGTATATC GTACTCGGAC 101 AGACCATGCT CAGAATCAAC CTGCACGCCT TTGTGTTAGG TTCGCTGATT 151 GCCGTCGTGG TGTGGTATTT CTTGTTTAAA TTCATTATCG GgGgTACTCA 201 ATATCCCCGA AAAGATGCAG CGTTTCGGTT CGGCnCGTAA AGGCCkCAAG 251 ssCGsGCTTG CCTTGAACAA GGCGGGTTTG GCGTATTTTG AAGGGCGTTT 301 TGAAAAGGCG GAACTAGAAG CCTCACGCGT GTTGGTCAAC AAAGtAGGCC 351 GajGAGACAAC CGGACTTTGG CATTGATGCT GrGCGCGCAC GCCGCCGGAC 401 AGATGGAAAA CATCGAssTG CGCGACCGTT ATCTTGCGGA AATCGCCAAA 451 CTGCCGGAAA AACAGCAGCT TTCCCGTTAT CTTTTGTTGG CGGAATCGGC 501 GTTGAACCGG CGCGATTACG AAGCGGCGGA AGCCAATCTT CATGCGGCGG 551 CGAAGATGAA TGCCAACCTT ACGCGCCTCG TGCGTCTGCA .ATTCGTTAC 601 GCTTTCGACA GGGGCGACGC GTTGCAGGTT CTGGCAAAAA CCGAAAAACT 651 TTCCAAGGCG GGCGCGTTGG GCAAATCGGA AATGGAACGG TATCAAAATT 701 GGGCATATCC GTCGCCAGCT GGCGGATGCT GCCGATGCCG CCGCTTTGAA 751 AACCTGCCTG AAGCGGATTC CCGACAGCCT CAAAAACGGG GAATTGAGCG 801 TATCGGTTGC GGAAAAGTAC GAACGTTTGG GACTGTATGC CGATGCGGTC 851 AAATGGGTCA AACAGCATTA TCCGCAsAAC CGCCGCCCCG AGCTTTTGGA 901 AGCCTTTGTC GAAAGCGTGC GCTTTTTGGG CGAGCGCGAA CAGCAGAAAG 951 CCATCGATTT TGCCGATGCT TGGCTGAAAG AACAGCCCGA TAACGCGCTT 1001 CTGCTGATGT ATCTCGGTCG GCTCGCCTTC GGCCGCAAAC TTTGGGGCAA 1051 GGCAAAAGGC TACCTTGAAG CGAGCATTGC ATTAAAGCCG AGTATTTCCG 1101 CGCGTTTGGT TCTAACAAAG GTTTTCGACG AAATCGGAGA ACCGCAGAAG 1151 GCGGAGGCGC AC... This coπesponds to the amino acid sequence <SEQ ID 750; ORF100>: 1 MKTWWIWL FAAAVGLALA SGIYTGDVYI VLGQTMLRIN LHAFVLGSLI 51 AVWWYFLFK FIIGVLNIPE KMQRFGSARK GXKXXLALNK AGLAYFEGRF 101 EKAELEASRV LVNKVGRDNR TLALMLXAHA AGQMENIXXR DRYLAEIAKL 151 PEKQQLSRYL LLAESALNRR DYEAAEANLH AAAKMNANLT RLVRLXIRYA 201 FDRGDALQVL AKTEKLSKAG ALGKSEMERY QNWAYRRQLA DAADAAALKT 251 CLKRIPDSLK NGELSVSVAE KYERLGLYAD AVKWVKQHYP XNRRPELLEA 301 FVESVRFLGE REQQKAIDFA DAWLKEQPDN ALLLMYLGRL AFGRKLWGKA 351 KGYLEASIAL KPSISARLVL TKVFDEIGEP QKAEAH... Further work revealed the complete nucleotide sequence <SEQ ID 751>: 1 ATGAAAACGG TAGTCTGGAT TGTCGTCCTG TTTGCCGCCG CCGTCGGACT 51 GGCGCTGGCT TCGGGCATTT ACACCGGCGA CGTGTATATC GTACTCGGAC 101 AGACCATGCT CAGAATCAAC CTGCACGCCT TTGTGTTAGG TTCGCTGATT 151 GCCGTCGTGG TGTGGTATTT CTTGTTTAAA TTCATTATCG GCGTACTCAA 201 TATCCCCGAA AAGATGCAGC GTTTCGGTTC GGCGCGTAAA GGCCGCAAGG 251 CCGCGCTTGC CTTGAACAAG GCGGGTTTGG CGTATTTTGA AGGGCGTTTT 301 GAAAAGGCGG AACTAGAAGC CTCACGCGTG TTGGTCAACA AAGAGGCCGG 351 AGACAACCGG ACTTTGGCAT TGATGCTGGG CGCGCACGCC GCCGGACAGA 401 TGGAAAACAT CGAGCTGCGC GACCGTTATC TTGCGGAAAT CGCCAAACTG 451 CCGGAAAAAC AGCAGCTTTC CCGTTATCTT TTGTTGGCGG AATCGGCGTT 501 GAACCGGCGC GATTACGAAG CGGCGGAAGC CAATCTTCAT GCGGCGGCGA 551 AGATGAATGC CAACCTTACG CGCCTCGTGC GTCTGCAACT TCGTTACGCT 601 TTCGACAGGG GCGACGCGTT GCAGGTTCTG GCAAAAACCG AAAAACTTTC 651 CAAGGCGGGC GCGTTGGGCA AATCGGAAAT GGAACGGTAT CAAAATTGGG 701 CATACCGCCG CCAGCTGGCG GATGCTGCCG ATGCCGCCGC TTTGAAAACC 751 TGCCTGAAGC GGATTCCCGA CAGCCTCAAA AACGGGGAAT TGAGCGTATC 801 GGTTGCGGAA AAGTACGAAC GTTTGGGACT GTATGCCGAT GCGGTCAAAT 851 GGGTCAAACA GCATTATCCG CACAACCGCC GCCCCGAGCT TTTGGAAGCC 901 TTTGTCGAAA GCGTGCGCTT TTTGGGCGAG CGCGAACAGC AGAAAGCCAT 951 CGATTTTGCC GATGCTTGGC TGAAAGAACA GCCCGATAAC GCGCTTCTGC 1001 TGATGTATCT CGGTCGGCTC GCCTACGGCC GCAAACTTTG GGGCAAGGCA 1051 AAAGGCTACC TTGAAGCGAG CATTGCATTA AAGCCGAGTA TTTCCGCGCG 1101 TTTGGTTCTA GCAAAGGTTT TCGACGAAAT CGGAGAACCG CAGAAGGCGG 1151 AGGCGCAGCG CAACTTGGTT TTGGAAGCCG TCTCCGATGA CGAACGTCAC 1201 GCAGCGTTAG AGCAGCATAG CTGA This coπesponds to the amino acid sequence <SEQ ID 752; ORF100-1>: 1 MKTWWIWL FAAAVGLALA SGIYTGDVYI VLGQTMLRIN LHAFVLGSLI_ 51 AVWWYFLFK FIIGVLNIPE KMQRFGSARK GRKAALALNK AGLAYFEGRF 101 EKAELEASRV LVNKEAGDNR TLALMLGAHA AGQMENIELR DRYLAEIAKL 151 PEKQQLSRYL LLAESALNRR DYEAAEANLH AAAKMNANLT RLVRLQLRYA 201 FDRGDALQVL AKTEKLSKAG ALGKSEMERY QNWAYRRQLA DAADAAALKT 251 CLKRIPDSLK NGELSVSVAE KYERLGLYAD AVKWVKQHYP HNRRPELLEA 301 FVESVRFLGE REQQKAIDFA DAWLKEQPDN ALLLMYLGRL AYGRKLWGKA 351 KGYLEASIAL KPSISARLVL AKVFDEIGEP QKAEAQRNLV LEAVSDDERH 401 AALEQHS* Computer analysis of this amino acid sequence gave the following results: Homology with a predicted ORF from N. meningitidis (sfrain A) ORFIOO shows 93.5% identity over a 386aa overlap with an ORF (ORFlOOa) from strain A of N meningitidis: 10 20 30 40 50 60 orf100. pep MKTWWIWLFAAAVGLALASGIYTGDVYIVLGQTMLRIΝLHAFVLGSLIAWVWYFLFK I I II I I I I I II I I I I I I I I I II I I I I I I I I II I II I I I I I I I I II II I II I II I I I II orf100a MKTWWIWLFAAAXGLALASGIXTGDVYIVLGQTMLRIΝLHAFVLGSLIAVWWYFLFK 10 20 30 40 50 60 70 80 90 100 110 120 orf100. pep FIIGVLΝIPEKMQRFGSARKGXKXXLALΝKAGLAYFEGRFEKAELEASRVLVΝKVGRDΝR I I I I I I I I I I I I I I I I I I II I II I II II I II I II II II II I I I I I II II : I I I orf100a FIIGVLΝXPEKMQRFGSARKGRKAALALΝKAGLAYFEGRFEKAELEASRVLGΝKEAGDΝR 70 80 90 100 110 120 130 140 150 160 170 180 orf100. pep TLALMLXAHAAGQMEΝIXXRDRYLAEIAKLPEKQQLSRYLLLAESALΝRRDYEAAEAΝLH MMM I I I I II I I I I I I II II I I I I I I I I I I II I II I I I II II II I II II II I I I I orf100a TLALMLGAHAAGQMEΝIELRDRYLAEIAKLPEKQQLSRYLLLAESALΝRRDYEAAEAΝLH 130 140 150 160 170 180 190 200 210 220 230 240 orf100.pep AAAKMΝAΝLTRLVRLXIRYAFDRGDALQVLAKTEKLSKAGALGKSEMERYQΝWAYRRQLA II I II II I II II I II : I II II I II I I I I I II I II II II I II I I I I I II I II I I I I I orf100a AAAKMΝAΝLTRLVRLQLRYAFDRGDALQVLAKTEKXSKAGAXGKSEMERYQΝWAYRRQLX 190 200 210 220 230 240 250 260 270 280 290 300 orf100. pep DAADAAALKTCLKRIPDSLKΝGELSVSVAEKYERLGLYADAVKWVKQHYPXΝRRPELLEA I I I I I I I II II I I I I I I II I I I II I I II I II I II I M I I I I I I II II I I I I II I II I I I orf100a DAADAAALKTCLKRIPDSLKΝGELSVSVAEKYERLGLYADAVKWVKQHYPHΝRRPELLEA 250 260 270 280 290 300 310 320 330 340 350 360 orf100. pep FVESVRFLGEREQQKAIDFADAWLKEQPDNALLLMYLGRLAFGRKLWGKAKGYLEASIAL I II II I I I II M I I I I I I I II I II II I I II I I II I I I I I I : I II I II I II I II II I I I I orf100a FVESVRFLGERDQQKAIDFADAWLKEQPDNALLLXYLGRLAYGRKLWGKAKGYLEASIAL 310 320 330 340 350 360 370 380 orf100.pep KPSISARLVLTKVFDEIGEPQKAEAH I I I I I I I I II : I II I I I I II I I I I : orflOOa KPSISARLVLAKVFDETGEPQKAEAQRNLVLASVAEENRPSAETHX 370 380 390 400 The complete length ORFlOOa nucleotide sequence <SEQ ID 753> is: 1 ATGAAAACGG TAGTCTGGAT TGTCGTCCTG TTTGCCGCCG CNNTCGGGCT 51 GGCATTGGCG TCGGGCATTN ACACCGGCGA CGTGTATATC GTACTCGGAC 101 AGACCATGCT CAGAATCAAC CTGCACGCCT TTGTGTTAGG TTCGCTGATT 151 GCCGTCGTGG TGTGGTATTT CCTGTTCAAA TTCATCATCG GCGTACTCAA 201 TANCCCCGAA AAGATGCAGC GTTTCGGTTC GGCGCGTAAA GGCCGCAAGG 251 CCGCGCTTGC TTTGAACAAG GCGGGTTTGG CGTATTTTGA AGGGCGTTTT 301 GAAAAGGCGG AACTTGAAGC CTCGCGCGTA TTGGGAAACA AAGAGGCGGG 351 GGATAACCGG ACTTTGGCAT TGATGTTGGG CGCACATGCC GCCGGGCAGA 401 TGGAAAACAT CGAGCTGCGC GACCGTTATC TTGCGGAAAT CGCCAAACTG 451 CCGGAAAAGC AGCAGCTTTC CCGTTATCTT TTGTTGGCGG AATCGGCGTT 501 GAACCGGCGC GATTACGAAG CGGCGGAAGC CAATCTTCAT GCGGCGGCGA 551 AGATGAATGC CAACCTTACG CGCCTCGTGC GTCTGCAACT TCGTTACGCT 601 TTCGACAGGG GCGACGCGTT GCAGGTTCTG GCAAAAACCG AAAAANTTTC 651 CAAGGCGGGC GCGTNGGGCA AATCGGAAAT GGAACGGTAT CAAAATTGGG 701 CATACCGCCG CCAGCTGNCG GATGCTGCCG ATGCCGCCGC TTTGAAAACC 751 TGCCTGAAGC GGATTCCCGA CAGCCTCAAA AACGGGGAAT TGAGCGTATC 801 GGTTGCGGAA AAGTACGAAC GTTTGGGACT GTATGCCGAT GCGGTCAAAT 851 GGGTCAAACA GCATTATCCG CACAACCGCC GACCCGAACT TTTGGAAGCN 901 TTTGTCGAAA GCGTGCGCTT TTTGGGCGAA CGCGATCAGC AGAAAGCCAT 951 CGATTTTGCC GATGCTTGGC TGAAAGAACA GCCCGATAAT GCGCTTCTGC 1001 TGANGTATCT CGGTCGGCTC GCCTACGGCC GCAAACTTTG GGGCAAGGCA 1051 AAAGGCTACC TTGAAGCGAG CATTGCATTA AAGCCGAGTA TTTCCGCGCG 1101 TTTGGTTCTG GCAAAGGTTT TTGACGAAAC CGGAGAACCG CAGAAGGCGG 1151 AGGCGCAGCG CAACTTGGTT TTGGCAAGCG TTGCCGAGGA AAACCGNCCT 1201 TCCGCCGAAA CCCATTGA This encodes a protein having amino acid sequence <SEQ ID 754>: 1 MKTWWIWL FAAAXGLALA SGIXTGDVYI VLGQTMLRIN LHAFVLGSLI 51 AVWWYFLFK FIIGVLNXPE KMQRFGSARK GRKAALALNK AGLAYFEGRF 101 EKAELEASRV LGNKEAGDNR TLALMLGAHA AGQMENIELR DRYLAEIAKL 151 PEKQQLSRYL LLAESALNRR DYEAAEANLH AAAKMNANLT RLVRLQLRYA 201 FDRGDALQVL AKTEKXSKAG AXGKSEMERY QNWAYRRQLX DAADAAALKT 251 CLKRIPDSLK NGELSVSVAE KYERLGLYAD AVKWVKQHYP HNRRPELLEA 301 FVESVRFLGE RDQQKAIDFA DAWLKEQPDN ALLLXYLGRL AYGRKLWGKA 351 KGYLEASIAL KPSISARLVL AKVFDETGEP QKAEAQRNLV LASVAEENRP 401 SAETH* ORFlOOa and ORF 100-1 show 95.1% identity in 406 aa overlap: 10 20 30 40 50 60 orf100a . pep MKTWWIWLFAAAXGLALASGIXTGDVYIVLGQTMLRINLHAFVLGSLIAVWWYFLFK I I I I I I II I I I I I I I I I I I I I I II II II I I I I I I I II I I I I I I I I I II II I I I I I II I orf100-1 MKTWWIWLFAAAVGLALASGIYTGDVYIVLGQTMLRINLHAFVLGSLIAVWWYFLFK 10 20 30 40 50 60 70 80 90 100 110 120 orf100a . pep FIIGVLNXPEKMQRFGSARKGRKAALALNKAGLAYFEGRFEKAELEASRVLGNKEAGDNR II I I I I I II I I II I I I II I I II I I I I I II I I II II II I I I I II I I I II I I I I I I I II I orf100-1 FIIGVLNIPEKMQRFGSARKGRKAALALNKAGLAYFEGRFEKAELEASRVLVNKEAGDNR 70 80 90 100 110 120 130 140 150 160 170 180 orf100a . pep TLALMLGAHAAGQMENIELRDRYLAEIAKLPEKQQLSRYLLLAESALNRRDYEAAEANLH I I II I I I I I I I I I I II II I I II I I II II II I I I I II I I I II I II I I I I I I I I I I I I I II I orf100-1 TLALMLGAHAAGQMENIELRDRYLAEIAKLPEKQQLSRYLLLAESALNRRDYEAAEANLH 130 140 150 160 170 180 190 200 210 220 230 240 orf100a . pep AAAKMNANLTRLVRLQLRYAFDRGDALQVLAKTEKXSKAGAXGKSEMERYQNWAYRRQLX I I I I I I I I II I II I I I II I I I I I II I I I I I I II II I I I I I I I I II II I I II I I I I II orf100-1 AAAKMNANLTRLVRLQLRYAFDRGDALQVLAKTEKLSKAGALGKSEMERYQNWAYRRQLA 190 200 210 220 230 240 250 260 270 280 290 300 orf100a . pep DAADAAALKTCLKRIPDSLKNGELSVSVAEKYERLGLYADAVKWVKQHYPHNRRPELLEA I I I II I I I II I II I I I II I I I I I II I I I I II II I I I I I I I II II I I I I I I I I II I I I I II orf100-1 DAADAAALKTCLKRIPDSLKNGELSVSVAEKYERLGLYADAVKWVKQHYPHNRRPELLEA 250 260 270 280 290 300 310 320 330 340 350 360 orf100a . pep FVESVRFLGERDQQKAIDFADAWLKEQPDNALLLXYLGRLAYGRKLWGKAKGYLEASIAL II I II II II I M I I I I II I I I I I II I I I I I I I II I II I I I I I II I I I II II I I II II I I orf100-1 FVESVRFLGEREQQKAIDFADAWLKEQPDNALLLMYLGRLAYGRKLWGKAKGYLEASIAL 310 320 330 340 350 360 370 380 390 400 orf100a. pep KPSISARLVLAKVFDETGEPQKAEAQRNLVLASVAEENRPSA-ETHX II I I II I I I I I II I I I I I I I I I II I I I I I I : I ::: : I M I I orf100-1 KPSISARLVLAKVFDEIGEPQKAEAQRNLVLEAVSDDERHAALEQHSX 370 380 390 400 Homology with a predicted ORF from N.εonorrhoeae ORFIOO shows 93.3% identity over a 386 aa overlap with a predicted ORF (ORFlOOng) from N.gonorrhoeae: orf100. pep MKTWWIWLFAAAVGLALASGIYTGDVYIVLGQTMLRINLHAFVLGSLIAVWWYFLFK 60 II I I II I I I I I I II I I I I I I I I I I I II I II II I I I I I I I I I II I II I I I I II I I I I II I I orflOOng MKTWWIWLFAAAVGLALASGIYTGDVYIVLGQTMLRINLHAFVLGSLIAVWWYFLFK 60 orf100.pep FIIGVLNIPEKMQRFGSARKGXKXXLALNKAGLAYFEGRFEKAELEASRVLVNKVGRDNR 120 II I II I I I I I : I : I I I I II I I II I II II II I I II I I I II I II I I II I II : II I orflOOng FIIGVLNIPENMRRSGSARKGRKAALALNKAGLAYFEGRFEKAELEASRVLGNKEAGDNR 120 orf100.pep TLALMLXAHAAGQMENIXXRDRYLAEIAKLPEKQQLSRYLLLAESALNRRDYEAAEANLH 180 II I II I I I I I I II I I I I I I I I I II I II II I I I I I I I I I II I I II I II I I I I I I I II I orflOOng TLALMLGAHAAGQMENIELRDRYLAEIAKLPEKQQLSRYLLLAESALNRRDYEAAEANLH 180 orf100.pep AAAKMNANLTRLVRLXIRYAFDRGDALQVLAKTEKLSKAGALGKSEMERYQNWAYRRQLA 240 I I I I I I I I I II I I I I : II I II I I II I I I I I I I I I II II II I I I II I I I I I II I I II M I orflOOng AAAKMNANLTRLVRLQLRYAFDRGDALQVLAKTEKLSKAGALGKSEMERYQNWAYRRQMA 240 orf100.pep DAADAAALKTCLKRIPDSLKNGELSVSVAEKYERLGLYADAVKWVKQHYPXNRRPELLEA 300 II I II I I I I I I I II I I I I II I I I I I I I I I I I II I II I I I I II I I I I I I I I I I II II I I I orflOOng DAADAAALKTCLKRIPDSLKNGELSVSVAEKYERLGLYADAVKWVKQHYPHNRRPELLEA 300 orf100. pep FVESVRFLGEREQQKAIDFADAWLKEQPDNALLLMYLGRLAFGRKLWGKAKGYLEASIAL 360 I I I I I I I I I II I I I II I I I I I l I I I I I I II I I I I II I I I: I II I I II I I I I II I I I I I orflOOng FVESVRFLGEREQQKAIDFADSWLKEQPDNALLLMYLGRLAYGRKLWGKAKGYLEASIAL 360 orf100. pep KPSISARLVLTKVFDEIGEPQKAEAH 386 II I I I I I I I: I II I I :: I I I II: orflOOng KPSIPARLVLAKVFDETAQSQKAEAQRNLVLASVAGENRPSAETR 405 The complete length ORFlOOng nucleotide sequence <SEQ ID 755> is: 1 ATGAAAACGG TAGTCTGGAT TGTTGTCCTG TTTGCCGCCG CCGTCGGACT 51 GGCGCTGGCT TCGGGCATTT ACACCGGCGA CGTGTATATC GTACTCGGAC 101 AGACCATGCT CAGAATCAAC CTGCACGCCT TTGTGTTAGG TTCGCTGATT 151 GCCGTCGTGG TGTGGTATTT CCTGTTTAAA TTCATCATCG GCGTACTCAA 201 TATCCCCGAA AATATGCGGC GTTCCGGTTC GGCGCGGAAA GGCCGCAAGG 251 CCGCGCTTGC CTTGAATAAG GCGGGTTTGG CGTATTTCGA AGGGCGTTTT 301 GAAAAGGCGG AACTCGAAGC CTCTCGAGTG TTGGGCAACA AAGAGGCCGG 351 AGACAACCGG ACTTTGGCAT TGATGCTGGG CGCGCACGCG GCAGGACAGA 401 TGGAAAATAT CGAGCTGCGC GACCGTTATC TTGCGGAAAT CGCCAAACTG 451 CCGGAAAAAC AGCAGCTTTC CCGCTATCTT CTGCTGGCGG AATCGGCGTT 501 AAACCGGCGC GATTACGAAG CGGCGGAAGC CAATCTTCAT GCGGCGGCGA 551 AGATGAATGC CAACCTTACG CGCCTCGTGC GTCTGCAACT TCGTTACGCC 601 TTCGATCGGG GCGATGCGTT GCAGGTTCTG GCAAAAaccG AAAAACTTTC 651 CAAGGCGGGC GCGTTGGGCA AATCGGAAAT GGAACGGTAT CAAAATTGGG 701 CATACCGCCG CCAGATGGCG GATGCTGCCG ATGCCGCCGC TTTGAAAACC 751 TGCCTGAAGC GGATTCCCGA CAGCCTCAAA AACGGGGAAT TGagcGTATC 801 GGTTGCGGAA AAGTACGAAC GTTTGGGACT GTATGCCGAT GCGGTCAAAT 851 GGGTCAAACA GCATTATCCG CACAACCGCC GCCCCGAGCT TTTGGAAGCC 901 TTTGTCGAAA GCGTGCGCTT TTTGGGCGAG CGCGAACAGC AGAAAGCCAT 951 CGATTTTGCC GATTCTTGGC TGAAAGAACA GCCCGATAAC GCGCTTCTGC 1001 TGATGTATCT CGGCCGGCTC GCCTACGGCC GCAAACTTTG GGGTAAGGCA 1051 AAAGGCTACC TTGAAGCGAG TATTGCACTG AAGCCGAGTA TTCCGGCGCG 1101 TTTGGTGTTG GCAAAGGTTT TTGACGAAAC CGCACAGTCG CAAAAAGCCG 1151 AAGCACAGCG CAACTTGGTT TTGGCAAGCG TTGCCGGGGA AAACCGCCCT 1201 TCCGCCGAAA CCCGTTGA This encodes a protem having amino acid sequence <SEQ LD 756>: 1 MKTWWIWL FAAAVGLALA SGIYTGDVYI VLGQTMLRIN LHAFVLGSLI 51 AVWWYFLFK FIIGVLNIPE NMRRSGSARK GRKAALALNK AGLAYFEGRF 101 EKAELEASRV LGNKEAGDNR TLALMLGAHA AGQMENIELR DRYLAEIAKL 151 PEKQQLSRYL LLAESALNRR DYEAAEANLH AAAKMNANLT RLVRLQLRYA 201 FDRGDALQVL AKTEKLSKAG ALGKSEMERY QNWAYRRQMA DAADAAALKT 251 CLKRIPDSLK NGELSVSVAE KYERLGLYAD AVKWVKQHYP HNRRPELLEA 301 FVESVRFLGE REQQKAIDFA DSWLKEQPDN ALLLMYLGRL AYGRKLWGKA 351 KGYLEASIAL KPSIPARLVL AKVFDETAQS QKAEAQRNLV LASVAGENRP 401 SAETR* ORFlOOng and ORFlOO-1 show 95.3% identity in 402 aa overlap: 10 20 30 40 50 60 orf100-1. pep MKTWWIWLFAAAVGLALASGIYTGDVYIVLGQTMLRINLHAFVLGSLIAVWWYFLFK I I I I I II I I II I I I I II I I II I II I II I I I I I I I II I I II I I II I I I II I I I II I I I I I I orflOOng MKTWWIWLFAAAVGLALASGIYTGDVYIVLGQTMLRINLHAFVLGSLIAVWWYFLFK 10 20 30 40 50 60 70 80 90 100 110 120 orf100-1. pep FIIGVLNIPEKMQRFGSARKGRKAALALNKAGLAYFEGRFEKAELEASRVLVNKEAGDNR I I I I II I I II : I : I I I I I II I II I II II I I I I I I I I I I I II II I I II I II I II II II I orflOOng FIIGVLNIPENMRRSGSARKGRKAALALNKAGLAYFEGRFEKAELEASRVLGNKEAGDNR 70 80 90 100 110 120 130 140 150 160 170 180 orf100-1. pep TLALMLGAHAAGQMENIELRDRYLAEIAKLPEKQQLSRYLLLAESALNRRDYEAAEANLH I I I I I I II I II II I I I I I I I I I I II I II II I I I I I I II I I I I I I II I II I II II I I I I II orflOOng TLALMLGAHAAGQMENIELRDRYLAEIAKLPEKQQLSRYLLLAESALNRRDYEAAEANLH 130 140 150 160 170 180 190 200 210 220 230 240 orf100-1. pep AAAKMNANLTRLVRLQLRYAFDRGDALQVLAKTEKLSKAGALGKSEMERYQNWAYRRQLA I I II I I I II I I I II II I I I II I I I I I I I I I I I II I I I II II II II II I I II I I I I I II: I orflOOng AAAKMNANLTRLVRLQLRYAFDRGDALQVLAKTEKLSKAGALGKSEMERYQNWAYRRQMA 190 200 210 220 230 240 250 260 270 280 290 300 orf100-1. pep DAADAAALKTCLKRIPDSLKNGELSVSVAEKYERLGLYADAVKWVKQHYPHNRRPELLEA II I I I II I II II I I I II II I I I I II I I I II I II I I I II II I I I I II I I I I II II I I I I I I orflOOng DAADAAALKTCLKRIPDSLKNGELSVSVAEKYERLGLYADAVKWVKQHYPHNRRPELLEA 250 260 270 280 290 300 310 320 330 340 350 360 orf100-1. pep FVESVRFLGEREQQKAIDFADAWLKEQPDNALLLMYLGRLAYGRKLWGKAKGYLEASIAL II I II II I I I I I I I I II I II I: I I II I I I II I II II I II I I I II I II I I II I I I I II II I orflOOng FVESVRFLGEREQQKAIDFADSWLKEQPDNALLLMYLGRLAYGRKLWGKAKGYLEASIAL 310 320 330 340 350 360 370 380 390 400 orf100-1.pep KPSISARLVLAKVFDEIGEPQKAEAQRNLVLEAVSDDERHAALEQHSX II I I II I II I I I II I :: I II I II I II I I : M :: I M orflOOn KPSIPARLVLAKVFDETAQSQKAEAQRNLVLASVAGENRPSAETRX 370 380 390 400 Based on this analysis, including the presence of a putative leader sequence, a putative transmembrane domain, and a RGD motif, it is predicted that the proteins from N. meningitidis .and N.gonorrhoeae, and their epitopes, could be useful antigens for vaccines or diagnostics, or for raising antibodies. Example 90 The following DNA sequence, believed to be complete, was identified in N. meningitidis <SEQ ID 757> 1 ATGATGTTTT CTTGGTTCAA GCTGTTTCAC TTGTTTTTTG TCATTTCGTG 51 GTTTGCAGGG CTGTTTTACC TGCCGAGGAT TTTCGTCAAT ATGGCGATGA 101 TTGATGTGCC GCGCGGCAAT CCCGAGTATG TGCGTCTGTC GGGCATGGCG 151 GTGCGGCTGT ACCGTTTTAT GTCGCCGTTG GGCTTCGGCG CGGTCGTGTT 201 CGGCGCGGCG ATACCGTTTG CCGCCGGCTG GTGGGGCAGC GGCTGGGTAC 251 ACGTCAAACT GTGTTTGGGC TTGATGCTCT TGGCTTACCA GTTGTATTGC 301 GGCGTGCTGC TGCGCCGTTT TCAGGATTAC AGCAATGCTT TTTCACACCG 351 CTGGTACCGC GTGTTCAACG AAATCCCCGT GCTGCTGATG GTTGCCGCGC 401 TGTATsTGGT CGTGTTCAAA CCGTTTTGA This coπesponds to the amino acid sequence <SEQ ED 758; ORF102>: 1 MMFSWFKLFH LFFVISWFAG LFYLPRIFVN MAMIDVPRGN PEYVRLSGMA 51 VRLYRFMSPL GFGAWFGAA IPFAAGWWGS GWVHVKLCLG LMLLAYQLYC 101 GVLLRRFQDY SNAFSHRWYR VFNEIPVLLM VAALYXWFK PF* Further work revealed the complete nucleotide sequence <SEQ ID 759>: 1 ATGATGTTTT CTTGGTTCAA GCTGTTTCAC TTGTTTTTTG TCATTTCGTG 51 GTTTGCAGGG CTGTTTTACC TGCCGAGGAT TTTCGTCAAT ATGGCGATGA 101 TTGATGTGCC GCGCGGCAAT CCCGAGTATG TGCGTCTGTC GGGCATGGCG 151 GTGCGGCTGT ACCGTTTTAT GTCGCCGTTG GGCTTCGGCG CGGTCGTGTT 201 CGGCGCGGCG ATACCGTTTG CCGCCGGCTG GTGGGGCAGC GGCTGGGTAC 251 ACGTCAAACT GTGTTTGGGC TTGATGCTCT TGGCTTACCA GTTGTATTGC 301 GGCGTGCTGC TGCGCCGTTT TCAGGATTAC AGCAATGCTT TTTCACACCG 351 CTGGTACCGC GTGTTCAACG AAATCCCCGT GCTGCTGATG GTTGCCGCGC 401 TGTATCTGGT CGTGTTCAAA CCGTTTTGA This coπesponds to the amino acid sequence <SEQ ID 760; ORF102-1>: 1 MMFSWFKLFH LFFVISWFAG LFYLPRIFVN MAMIDVPRGN PEYVRLSGMA 51 VRLYRFMSPL GFGAWFGAA IPFAAGWWGS GWVHVKLCLG LMLLAYQLYC 101 GVLLRRFQDY SNAFSHRWYR VFNEIPVLLM VAALYLWFK PF* Computer analysis of this amino acid sequence gave the following results: Homology with HP1484 hypothetical integral membrane protein of H. pylori (accession number AE000647) ORF102 and HP1484 show 33% aa identity in 143aa overlap: orfl02 3 FSWFKLFHLFFVISWFAGLFYLPRIFVNMAMIDVPRGNPEYVRLSGMAVRLYRFMSPLGF 62 F W K FH+ VISW A LFYLPR+FV A + V++ +LY F++ HP1484 8 FLWVKAFHVIAVISWMAALFYLPRLFVYHAENAHKKEFVGWQIQEK--KLYSFIASPAM 65 orfl02 63 GAWFGAAIPFAAG WWGSGWVHVKLCLGLMLLAYQLYCGVLLRRFQDYSNAFSHRWY 119 G + + + GW+H KL L ++LLAY YC +R + + R+Y HP1484 66 GFTLITGILMLLIEPTLFKSGGWLHAKLALWLLLAYHFYCKKCMRELEKDPTRRNARFY 125 orfl02 120 RVFNEIPXXXXXXXXXXXXFKPF 142 RVFNE P KPF HP1484 126 RVFNEAPTILMILIVILVWKPF 148 Homology with a predicted ORF from N.meninsitidis (strain Al ORF102 shows 99.3% identity over a 142aa overlap with an ORF (ORF102a) from strain A of N meningitidis: 10 20 30 40 50 60 orf102. pep MMFSWFKLFHLFFVISWFAGLFYLPRIFVΝMAMIDVPRGΝPEYVRLSGMAVRLYRFMSPL I II I I I I I I I II I I I II II II I I I I I II I I I I I II I I I I I I I I I II I I I I I I I I II I I II orf102a MMFSWFKLFHLFFVISWFAGLFYLPRIFVΝMAMIDVPRGΝPEYVRLSGMAVRLYRFMSPL 10 20 30 40 50 60 70 80 90 100 110 120 orf102. pep GFGAWFGAAIPFAAGWWGSGWVHVKLCLGLMLLAYQLYCGVLLRRFQDYSΝAFSHRWYR I I I II I I II I I I I I I I I I I II I II I I I I I I I I I I I I I I I I I I I I II II II I I II I I I I I I orf102a GFGAWFGAAIPFAAGWWGSGWVHVKLCLGLMLLAYQLYCGVLLRRFQDYSΝAFSHRWYR 70 80 90 100 110 120 130 140 orfl02.pep VFΝEIPVLLMVAALYXWFKPFX I I II II II II I II II II II I II orfl02a VFΝEIPVLLMVAALYLWFKPFX 130 140 The complete length ORF 102a nucleotide sequence <SEQ ID 761 > is: 1 ATGATGTTTT CTTGGTTCAA GCTGTTTCAC TTGTTTTTTG TCATTTCGTG 51 GTTTGCAGGG CTGTTTTACC TGCCGAGGAT TTTCGTCAAT ATGGCGATGA 101 TTGATGTGCC GCGCGGCAAT CCCGAGTATG TGCGTCTGTC GGGCATGGCG 151 GTGCGGCTGT ACCGTTTTAT GTCGCCGTTG GGCTTCGGCG CGGTCGTGTT 201 CGGCGCGGCG ATACCGTTTG CCGCCGGCTG GTGGGGCAGC GGCTGGGTAC 251 ACGTCAAACT GTGTTTGGGC TTGATGCTCT TGGCTTACCA GTTGTATTGC 301 GGCGTGCTGC TGCGCCGTTT TCAGGATTAC AGCAATGCTT TTTCACACCG 351 CTGGTACCGC GTGTTCAACG AAATCCCCGT GCTGCTGATG GTTGCCGCGC 401 TGTATCTGGT CGTGTTCAAA CCGTTTTGA This encodes a protein having amino acid sequence <SEQ ED 762>: 1 MMFSWFKLFH LFFVISWFAG LFYLPRIFVN MAMIDVPRGN PEYVRLSGMA 51 VRLYRFMSPL GFGAWFGAA IPFAAGWWGS GWVHVKLCLG LMLLAYQLYC 101 GVLLRRFQDY SNAFSHRWYR VFNEIPVLLM VAALYLWFK PF* ORF 102a and ORF 102-1 show complete identity in 142 aa overlap: 10 20 30 40 50 60 orf102 . pep MMFSWFKLFHLFFVISWFAGLFYLPRIFVNMAMIDVPRGNPEYVRLSGMAVRLYRFMSPL I II II II I I I I I I II I I I I I I I II I I II I I I II I II I I I I I I I I I II I II I II II II II I orf102-1 MMFSWFKLFHLFFVISWFAGLFYLPRIFVNMAMIDVPRGNPEYVRLSGMAVRLYRFMSPL 10 20 30 40 50 60 70 80 90 100 110 120 orf102a . pep GFGAWFGAAIPFAAGWWGSGWVHVKLCLGLMLLAYQLYCGVLLRRFQDYSNAFSHRWYR I I I I I I I I II I I I I I I II I II I I II II II II I I II II I I I I II I II I I I II II I I I I I II orf102-1 GFGAWFGAAIPFAAGWWGSGWVHVKLCLGLMLLAYQLYCGVLLRRFQDYSNAFSHRWYR 70 80 90 100 110 120 130 140 orf102a. pep VFNEIPVLLMVAALYLWFKPFX I I I II II II I I I I I I I I I I I I I I orfl02-l VFNEIPVLLMVAALYLWFKPFX 130 140 Homology with a predicted ORF from N.εonorrhoeae ORF 102 shows 97.9% identity over a 142 aa overiap with a predicted ORF (ORF102ng) from N. gonorrhoeae: orf102. pep MMFSWFKLFHLFFVISWFAGLFYLPRIFVNMAMIDVPRGNPEYVRLSGMAVRLYRFMSPL 60 I II I II II I II II II II I II I I I I I II I I II I I I I : I I I I I II I I I I I I I I I I I II I II I orfl02ng MMFSWFKLFHLFFVISWFAGLFYLPRIFVNMAMIDAPRGNPEYVRLSGMAVRLYRFMSPL 60 orf102.pep GFGAWFGAAIPFAAGWWGSGWVHVKLCLGLMLLAYQLYCGVLLRRFQDYSNAFSHRWYR 120 II I II I I I I I I II II I I II II I I I II I I II I I I I II I I I II I II II I II I II I I II II I orfl02ng GFGAWFGAAIPFAAGRWGSGWVHVKLCLGLMLLAYQLYCGVLLRRFQDYSNAFSHRWYR 120 orf102. pep VFNEIPVLLMVAALYXWFKPF 142 II I II I I I II I II II I I II II orfl02ng VFNEIPVLLMVAALYLWFKPF 142 The complete length ORF102ng nucleotide sequence <SEQ LD 763> is: 1 ATGATGTTTT CTTGGTTCAA GCTGTTTCAC TTGTTTTTTG TCATTTCGTG 51 GTTTGCAGGG CTGTTTTACC TGCCGAGGAT TTTCGTCAAT ATGGCGATGA 101 TTGATGCGCC GCGCGGCAAT CCCGAGTATG TGCGCCTGTC GGGGATGGCG 151 GTGCGGTTGT ACCGTTTTAT GTCGCCTTTG GGTTTCGGCG CGGTCGTGTT 201 CGGCGCGGCG ATACCGTTTG CCGCcggccg GTGGGGCagc ggctggGTTC 251 ACGTCAAACT GTGTTTGGGC TTGATGCTCT TGGCTTATCA GTTGTATTGC 301 GGCGTGCTGC TGCGCCGTTT TCAGGATTAC AGCAATGCTT TTTCACACCG 351 CTGGTACCGC GTGTTCAAcg aAATCCCCGT GCTGCTGATG GTTGCCGCGC 401 TGTATCTGGT CGTGTTCAAA CCGTTTTGA This encodes a protein having amino acid sequence <SEQ ED 764>: 1 MMFSWFKLFH LFFVISWFAG LFYLPRIFVN MAMIDAPRGN PEYVRLSGMA 51 VRLYRFMSPL GFGAWFGAA IPFAAGRWGS GWVHVKLCLG LMLLAYQLYC 101 GVLLRRFQDY SNAFSHRWYR VFNEIPVLLM VAALYL FK PF* ORF102ng and ORF102-1 show 98.6% identity in 142 aa overlap: 10 20 30 40 50 60 orf102-1. pep MMFSWFKLFHLFFVISWFAGLFYLPRIFVNMAMIDVPRGNPEYVRLSGMAVRLYRFMSPL I I II II I II I I I I I I I II I II I I I I I I II I I I I I I : I II I I I I I I I I I II I II I I I I I I I orfl02ng MMFSWFKLFHLFFVISWFAGLFYLPRIFVNMAMIDAPRGNPEYVRLSGMAVRLYRFMSPL 10 20 30 40 50 60 70 80 90 100 110 120 orf102-1. pep GFGAWFGAAIPFAAGWWGSGWVHVKLCLGLMLLAYQLYCGVLLRRFQDYSNAFSHRWYR I I II II I I II I I I I I I II I I I I I I I I I I I I I I I I I I I I II II II I II I II I I II II II I orfl02ng GFGAWFGAAIPFAAGRWGSGWVHVKLCLGLMLLAYQLYCGVLLRRFQDYSNAFSHRWYR 70 80 90 100 110 120 130 140 orfl02-l.pep VFNEIPVLLMVAALYLWFKPFX I I I II II I II I II I I I I I I I I II orfl02ng VFNEIPVLLMVAALYLWFKPFX 130 140 In addition, ORF102ng shows significant homology to a membrane protein from H.pylori: gi 12314656 (AE000647) conserved hypothetical integral membrane protein [Helicobacter pylori] Length = 148 Score = 79.2 bits (192), Expect = le-14 Identities = 50/147 (34%), Positives = 68/147 (46%), Gaps = 13/147 (8%) Query: 3 FSWFKLFHLFFVISWFAGLFYLPRIFVNMAMIDAPRGNPEYVRLSGMAVRLYRFMSPLGF 62 F W K FH+ VISW A LFYLPR+FV A + V++ +LY F++ Sbjct: 8 FLWVKAFHVIAVISWMAALFYLPRLFVYHAENAHKKEFVGWQIQEK—KLYSFIASPAM 65 Query : 63 GAWFGAAIP FAAGRWGSGWVHVKLCLGLMLLAYQLYCGVLLRRFQDYSNAFS 115 G + + F +G GW+H KL L ++LLAY YC +R + + Sbj ct : 66 GFTLITGILMLLIEPTLFKSG GWLHAKLALWLLLAYHFYCKKCMRELEKDPTRRN 121 Query: 116 HRWYRVFNEIPXXXXXXXXXXXXFKPF 142 R+YRVFNE P KPF Sbjct: 122 ARFYRVFNEAPTILMILIVILVWKPF 148 Based on this analysis, it is predicted that these proteins from N. meningitidis and N.gonorrhoeae, and their epitopes, could be useful antigens for vaccines or diagnostics, or for raising antibodies. Example 91 The following partial DNA sequence was identified in N. meningitidis <SEQ ID 765>: 1 ATGGCAAAAA TGATGAAATG GGCGGCTGTT GCGGCGGTCG CGGCGGCAGC 51 GGTTTGGGGC GGATGGTCTT AACTGAAGCC CGAGCCGCAC GTGCTTGATA 101 TTACGGAAAC GGTCAGGCGC GGC // //.. ATTTCGTTTA CGATTTTGTC CGAACCGGAT ACGCCGATTA AGGCGAAGCT 51 CGACAGCGTC GACCCCGGGC TGACCACGAT GTCGTCGGGC GGTTACAACA 101 GCAGTACGGA TACGGCTTCC AATGCGGTCT ACTATTATGC CCGTTCGTTT 151 GTGCCGAATC CGGACGGCAA ACTCGCCACG GGGATGACGA CGCAGAAT C 201 GGTTGAAATC GACGGCGTGA AAAATGTGCT GATTATTCCG TCGCTGACCG 251 TGAAAAATCG CGGCGGCAAG GCGTTTGTGC GCGTGTTGGG TGCGGACGGC 301 AAGGCGGCGG AACGCGAAAT CCGGACCGGT ATGAGAGACA GTATGAATAC 351 CGAAGTAAAA AGCGGGTTGA AAGAGGGGGA CAAAGTGGTC ATCTCCGAAA 401 TAACCGCCGC CGAGCAACAG GAAAGCGGCG AACGCGCCCT AGGCGGCCCG 451 CCGCGCCGAT AA This coπesponds to the amino acid sequence <SEQ ID 766; ORF85>: 1 MAKMMKWAAV AAVAAAAVWG GWS.LKPEPH VLDITETVRR G 101 151 201 i SFΓILSEPDT 251 PIKAKLDSVD PGLTTMSSGG YNSSTDTASN AVYYYARSFV PNPDGKLATG 301 MTTQNTVEID GVKNVLIIPS LTVKNRGGKA FVRVLGADGK AAEREIRTGM 351 RDSMNTEVKS GLKEGDKWI SEITAAEQQE SGERALGGPP RR* Further work revealed the further partial nucleotide sequence <SEQ ID 767>: 1 .. GTATCGGTCG GCGCGCAGGC ATCGGGGCAG ATTAAGATAC TTTATGTCAA 51 ACTCGGGCAA CAGGTTAAAA AGGGCGATTT GATTGCGGAA ATCAATTCGA 101 CCTCGCAGAC CAATACGCTC AATACGGAAA AATCCAAGTT GGAAACGTAT 151 CAGGCGAAGC TGGTGTCGGC ACAGATTGCA TTGGGCAGCG CGGAGAAGAA 201 ATATAAGCGT CAGGCGGCGT TATGGAAGGA AAACGCGACT TCCAAAGAGG 251 ATTTGGAAAG CGCGCAGGAT GCGTTTGCCG CCGCCAAAGC CAATGTTGCC 301 GAGCTGAAGG CTTTAATCAG ACAGAGCAAA ATTTCCATCA ATACCGCCGA 351 GTCGGAATTG GGCTACACGC GCATTACCGC AACGATGGAC GGCACGGTGG 401 TGGCGATTCT CGTGGAAGAG GGGCAGACTG TGAACGCGGC GCAGTCTACG 451 CCGACGATTG TCCAATTGGC GAATCTGGAT ATGATGTTGA ACAAAATGCA 501 GATTGCCGAG GGCGATATTA CCAAGGTGAA GGCGGGGCAG GATATTTCGT 551 TTACGATTTT GTCCGAACCG GATACGCCGA TTAAGGCGAA GCTCGACAGC 601 GTCGACCCCG GGCTGACCAC GATGTCGTCG GGCGGTTACA ACAGCAGTAC 651 GGATACGGCT TCCAATGCGG TCTACTATTA TGCCCGTTCG TTTGTGCCGA 701 ATCCGGACGG CAAACTCGCC ACGGGGATGA CGACGCAGAA TACGGTTGAA 751 ATCGACGGCG TGAAAAATGT GCTGATTATT CCGTCGCTGA CCGTGAAAAA 801 TCGCGGCGGC AAGGCGTTTG TGCGCGTGTT GGGTGCGGAC GGCAAGGCGG 851 CGGAACGCGA AATCCGGACC GGTATGAGAG ACAGTATGAA TACCGAAGTA 901 AAAAGCGGGT TGAAAGAGGG GGACAAAGTG GTCATCTCCG AAATAACCGC 951 CGCCGAGCAA CAGGAAAGCG GCGAACGCGC CCTAGGCGGC CCGCCGCGCC 1001 GATAA This coπesponds to the amino acid sequence <SEQ ID 768; ORF85-1 > • 1 ..VSVGAQASGQ IKILYVKLGQ QVKKGDLIAE INSTSQTNTL NTEKSKLETY 51 QAKLVSAQIA LGSAEKKYKR .QAALWKENAT SKEDLESAQD AFAAAKANVA 101 ELKALIRQSK ISINTAESEL GYTRITATMD GTWAILVEE GQTVNAAQST 151 PTIVQLANLD MMLNKMQIAE GDITKVKAGQ DISFTILSEP DTPIKAKLDS 201 VDPGLTTMSS GGYNSSTDTA SNAVYYYARS FVPNPDGKLA TGMTTQNTVE 251 IDGVKNVLII PSLTVKNRGG KAFVRVLGAD GKAAEREIRT GMRDSMNTEV 301 KSGLKEGDKV VISEITAAEQ QESGERALGG PPRR* Computer analysis of this amino acid sequence gave the following results: Homology with a predicted ORF from N.meninsitidis (sfrain A) ORF85 shows 87.8% identity over a 41aa overlap and 99.3% identity over a 153aa overlap with an ORF (ORF85a) from sfrain A of N. meningitidis: 10 20 30 40 orf85. pep MAKMMKWAAVAAVAAAAVWGGWS-LKPEPHVLDITETVRRG II I I I II II II II I I I I I I I I II Mill:: II II I I I I orf85a MAKMMKWAAVAAVAAAAVWGGWSYLKPEPQAAYITETVRRGDISRTVSATGEISPSΝLVS 10 20 30 40 50 60 // 80 90 100 orf85. pep ISFTILSEPDTPIKAKLDSVDPGLTTMSSG I I I I I I I I I I I I I I I I I II I I I I II II I I I orf85a TIVQLAΝLDMMLΝKMQIAEGDITKVKAGQDISFTILSEPDTPIKAKLDSVDPGLTTMSSG 210 220 230 240 250 260 110 120 130 140 150 160 orf85.pep GYΝSSTDTASΝAVYYYARSFVPΝPDGKLATGMTTQΝTVEIDGVKΝVLIIPSLTVKΝRGGK I II I I I I I I I II I I I I II I I II I II I I I I I I I II I II I I I I II I I I I I I I I I I I I I I I I : orf85a GYΝSSTDTASΝAVYYYARSFVPΝPDGKLATGMTTQΝTVEIDGVKΝVLIIPSLTVKΝRGGR 270 280 290 300 310 320 170 180 190 200 210 220 orf85. pep AFVRVLGADGKAAEREIRTGMRDSMΝTEVKSGLKEGDKWISEITAAEQQESGERALGGP I I II II II II I I I I I I I I I I I II I I II I I I I II I I I I I II II II II I I I I I II I II I I II orf85a AFVRVLGADGKAAEREIRTGMRDSMΝTEVKSGLKEGDKWISEITAAEQQESGERALGGP 330 340 350 360 370 380 230 orf85. pep PRRX I I II orf85a PRRX 390 The complete length ORF85a nucleotide sequence <SEQ ID 769> is: 1 ATGGCAAAAA TGATGAAATG GGCGGCTGTT GCGGCGGTCG CGGCGGCAGC 51 GGTTTGGGGC GGATGGTCTT ATCTGAAGCC CGAGCCGCAG GCTGCTTATA 101 TTACGGAAAC GGTCAGGCGC GGCGACATCA GCCGGACGGT TTCTGCAACA 151 GGGGAGATTT CGCCGTCCAA CCTGGTATCG GTCGGCGCGC AGGCATCGGG 201 GCAGATTAAG AAACTTTATG TCAAACTCGG GCAACAGGTT AAAAAGGGCG 251 ATTTGATTGC GGAAATCAAT TCGACCTCGC AGACCAATAC GCTCAATACG 301 GAAAAATCCA AATTGGAAAC GTATCAGGCG AAGCTGGTGT CGGCACAGAT 351 TGCATTGGGC AGCGCGGAGA AGAAATATAA GCGTCAGGCG GCGTTGTGGA 401 AGGATGATGC GACCGCTAAA GAAGATTTGG AAAGCGCACA GGATGCGCTT 451 GCCGCCGCCA AAGCCAATGT TGCCGAGCTG AAGGCTCTAA TCAGACAGAG 501 CAAAATTTCC ATCAATACCG CCGAGTCGGA ATTGGGCTAC ACGCGCATTA 551 CCGCAACGAT GGACGGCACG GTGGTGGCGA TTCTCGTGGA AGAGGGGCAG 601 ACTGTGAACG CGGCGCAGTC TACGCCGACG ATTGTCCAAT TGGCGAATCT 651 GGATATGATG TTGAACAAAA TGCAGATTGC CGAGGGCGAT ATTACCAAGG 701 TGAAGGCGGG GCAGGATATT TCGTTTACGA TTTTGTCCGA ACCGGATACG 751 CCGATTAAGG CGAAGCTCGA CAGCGTCGAC CCCGGGCTGA CCACGATGTC 801 GTCGGGCGGC TACAACAGCA GTACGGATAC GGCTTCCAAT GCGGTCTACT 851 ATTATGCCCG TTCGTTTGTG CCGAATCCGG ACGGCAAACT CGCCACGGGG 901 ATGACGACGC AGAATACGGT TGAAATCGAC GGTGTGAAAA ATGTGCTGAT 951 TATTCCGTCG CTGACCGTGA AAAATCGCGG CGGCAGGGCG TTTGTGCGCG 1001 TGTTGGGTGC AGACGGCAAG GCGGCGGAAC GCGAAATCCG GACCGGTATG 1051 AGAGACAGTA TGAATACCGA AGTAAAAAGC GGGTTGAAAG AGGGGGACAA 1101 AGTGGTCATC TCCGAAATAA CCGCCGCCGA GCAGCAGGAA AGCGGCGAAC 1151 GCGCCCTAGG CGGCCCGCCG CGCCGATAA This encodes a protein having amino acid sequence <SEQ ID 770>: 1 MAKMMKWAAV AAVAAAAVWG GWSYLKPEPQ AAYITETVRR GDISRTVSAT 51 GEISPSΝLVS VGAQASGQIK KLYVKLGQQV KKGDLIAEIΝ STSQTΝTLΝT 101 EKSKLETYQA KLVSAQIALG SAEKKYKRQA ALWKDDATAK EDLESAQDAL 151 AAAKAΝVAEL KALIRQSKIS IΝTAESELGY TRITATMDGT WAILVEEGQ 201 TVΝAAQSTPT IVQLAΝLDMM LΝKMQIAEGD ITKVKAGQDI SFTILSEPDT 251 PIKAKLDSVD PGLTTMSSGG YΝSSTDTASΝ AVYYYARSFV PΝPDGKLATG 301 MTTQΝTVEID GVKΝVLIIPS LTVKΝRGGRA FVRVLGADGK AAEREIRTGM 351 RDSMNTEVKS GLKEGDKWI SEITAAEQQE SGERALGGPP RR* ORF85a and ORF85-1 show 98.2% identity in 334 aa overlap: 30 40 50 60 70 80 orf 85a . pep PQAAYITETVRRGDISRTVSATGEISPSNLVSVGAQASGQIKKLYVKLGQQVKKGDLIAE I I I I I I I I I I I I I I I I I I I I I I I I I I I I I orf 85-1 VSVGAQASGQIKILYVKLGQQVKKGDLIAE 10 20 30 90 100 110 120 130 140 orf85a. pep INSTSQTNTLNTEKSKLETYQAKLVSAQIALGSAEKKYKRQAALWKDDATAKEDLESAQD I II I I II I II I II II II II I I II I II I I I I I II I I I II I I I I II I I :: I I: I I II II II I orf85-1 INSTSQTNTLNTEKSKLETYQAKLVSAQIALGSAEKKYKRQAALWKENATSKEDLESAQD 40 50 60 70 80 90 150 160 170 180 190 200 orf85a . pep ALAAAKANVAELKALIRQSKISINTAESELGYTRITATMDGTWAILVEEGQTVNAAQST I : I II I II I II I I I II II I II I I I I I II I II I I II I I I I I I I I II I I I I I II I II II II I orf85-1 AFAAAKANVAELKALIRQSKISINTAESELGYTRITATMDGTWAILVEEGQTVNAAQST 100 110 120 130 140 150 210 220 230 240 250 260 orf85a. pep PTIVQLANLDMMLNKMQIAEGDITKVKAGQDISFTILSEPDTPIKAKLDSVDPGLTTMSS II I I II I II I I I II II I II I II I I II I I I I II II I II I I I I I I I I I II I I I I II II II I I orf85-1 PTIVQLANLDMMLNKMQIAEGDITKVKAGQDISFTILSEPDTPIKAKLDSVDPGLTTMSS 160 170 180 190 200 210 270 280 290 300 310 320 orf85a. pep GGYNSSTDTASNAVYYYARSFVPNPDGKLATGMTTQNTVEIDGVKNVLIIPSLTVKNRGG I I II I II I II I I I I II II I II I I II I II II I II I I II I I I I II I I I I I II I I II I I I I II orf85-1 GGYNSSTDTASNAVYYYARSFVPNPDGKLATGMTTQNTVEIDGVKNVLIIPSLTVKNRGG 220 230 240 250 260 270 330 340 350 360 370 380 orf85a . pep RAFVRVLGADGKAAEREIRTGMRDSMNTEVKSGLKEGDKWISEITAAEQQESGERALGG : M I M I M I II II II I II I I I II I II I I II II I II II I I I I I I II I I II II I I II II II orf85-1 KAFVRVLGADGKAAEREIRTGMRDSMNTEVKSGLKEGDKWISEITAAEQQESGERALGG 280 290 300 310 320 330 390 orf85a. pep PPRRX II I I I orf85-l PPRRX Figure 19D shows plots of hydrophilicity, antigenic index, and AMPHI regions for ORF85a.. Homology with a predicted ORF from N.εonorrhoeae ORF85 shows a high degree of identity with a predicted ORF (ORF85ng) from N.gonorrhoeae: ORF85 1 MAKMMKWAAVAAVAAAAVWGGWS . LKPEPHVLDITETVRRG 40 II II I II I II I II I II I I II I II MM : I II : II I I ORF85ng 1 MAKMMKWAAVAAVAAAAVWGGWSYLKPEPQAAYITEAVRRGDISRTVSAT 50 ORF85 ISFTILSEPDT 250 I I I I I I I I I I I ORF85ng 201 TVNAAQSTPTIVQLANLDMMLNKMQIAEGDITKVKAGQDISFTILSEPDT 250 ORF85 251 PIKAKLDSVDPGLTTMSSGGYNSSTDTASNAVYYYARSFVPNPDGKLATG 300 I I I I I II I II I II I II I I II I I I I I II II I I I I I I I I I I I I I I I I I I I I I ORF85ng 251 PIKAKLDSVDPGLTTMSSGGYNSSTDTASNAVYYYARSFVPNPDGKLATG 300 ORF85 301 MTTQNTVEIDGVKNVLIIPSLTVKNRGGKAFVRVLGADGKAAEREIRTGM 350 I II I I II I II I II I II : I II I I I II I I II II I I I I I II I I I I I I I I I II ORF85ng 301 MTTQNTVEIDGVKNVLLIPSLTVKNRGGKAFVRVLGADGKAVEREIRTGM 350 ORF85 152 RDSMNTEVKSGLKEGDKWISEITAAEQQESGERALGGPPRR 393 : II I II I I I I I I I I II I I II I I I I I II I I I II I II II I I I I I 0RF85ng 351 KDSMNTEVKSGLKEGDKWISEITAAEQQESGERALGGPPRR 393 The complete length ORF85ng nucleotide sequence <SEQ ID 771 > is: 1 ATGGCAAAAA TGATGAAATG GGCGGCTGTT GCGGCGGTCG CGGCGGCaac 51 GGTTTGGGGC GGATGGTCTT ATCTGAAGCC CGAACCGCAG GCTGCTTATA 101 TTACGGAaac ggTCAGGCGC GGCGATATCA GCCGGACGGT TTCCGCGACG 151 GgcgAGATTT CGCCGTCCAA CCTGGTATCG GTCGGCGCGC AGGCTTCGGG 201 GCAGATTAAA AAGCTTTATG TCAAACTCGG GCAACAGGTC AAAAAGGGCG 251 ATTTGATTGC GGAAATCAAT TCGACCACGC AGACCAACAC GATCGATATG 301 GAAAAATCCA AATTGGAAAC GTATCAGGCG AAGCTGGTGT CGGCACAGAT 351 TGCATTGGGC AGCGCGGAGA AGAAATATAA GCGTCAGGCG GCGTTGTGGA 401 AGGATGATGC GACCTCTAAA GAAGATTTGG AAAGCGCGCA GGATGCGCTT 451 GCCGCCGCCA AAGCCAATGT TGCCGAGTTG AAGGCTTTAA TCAGACAGAG 501 CAAAATTTCC ATCAATACCG CCGAGTCGGA TTTGGGCTAC ACGCGCATTA 551 CCGCGACGAT GGACGGCACG GTGGTGGCGA TTCCCGTGGA AGAGGGGCAG 601 ACTGTGAACG CGGCGCAGTC TACGCCGACG ATTGTCCAAT TGGCGAATCT 651 GGATATGATG TTGAACAAAA TGCAGATTGC CGAGGGCGAT ATTACCAAGG 701 TGAAGGCGGG GCAGGATATT TCGTTTACGA TTTTGTCCGA ACCGGATACG 751 CCGATTAAGG CGAAGCTCGA CAGCGTCGAC CCCGGGCTGA CCACGATGTC 801 GTCGGGCGGC TACAACAGCA GTACGGATAC GGCTTCCAAT GCGGTCTATT 851 ATTATGCCCG TTCGTTTGTG CCGAATCCGG ACGGCAAACT CGCCACGGGG 901 ATGACGACGC AGAATACGGT TGAAATCGAC GGTGTGAAAA ATGTGTTGCT 951 TATTCCGTCG CTGACCGTGA AAAATCGCGG CGGCAAGGCG TTCGTACGCG 1001 TGTTGGGTGC GGACGGCAAG GCAGTGGAAC GCGAAATCCG GACCGGTATG 1051 AAAGACAGTA TGAATACCGA AGTGAAAAGC GGGTTGAAAG AGGGGGACAA 1101 AGTGGTCATC TCCGAAATAA CCGCCGCCGA GCAGCAGGAA AGCGGCGAAC 1151 GCGCCCTAGG CGGCCCGCCG CGCCGATAA This encodes a protein having amino acid sequence <SEQ ID 772>: 1 MAKMMKWAAV AAVAAAAVWG GWSYLKPEPQ AAYITEAVRR_ GDISRTVSAT 51 GEISPSNLVS VGAQASGQIK KLYVKLGQQV KKGDLIAEIN STTQTNTIDM 101 EKSKLETYQA KLVSAQIALG SAEKKYKRQA ALWKDDATSK EDLESAQDAL 151 AAAKANVAEL KALIRQSKIS INTAESDLGY TRITATMDGT WAIPVEEGQ 201 TVNAAQSTPT IVQLANLDMM LNKMQIAEGD ITKVKAGQDI SFTILΞEPDT 251 PIKAKLDSVD PGLTTMSSGG YNSSTDTASN AVYYYARSFV PNPDGKLATG 301 MTTQNTVEID GVKNVLLIPS LTVKNRGGKA FVRVLGADGK AVEREIRTGM 351 KDSMNTEVKS GLKEGDKWI SEITAAEQQE SGERALGGPP RR* ORF85ng and ORF85-1 show 96.1% identity in 334 aa overlap: 30 40 50 60 70 80 orf85ng PQAAYITETVRRGDISRTVSATGEISPSNLVSVGAQASGQIKKLYVKLGQQVKKGDLIAE I II I I II II I I I I II II I I II II II I I I I or 85-1 VSVGAQASGQIKILYVKLGQQVKKGDLIAE 10 20 30 90 100 110 120 130 140 orf85ng INSTTQTNTIDMEKSKLETYQAKLVSAQIALGSAEKKYKRQAALWKDDATSKEDLESAQD II I I : I I I I:: I I I II II I I II I II I I II I I I I I II I I II I II II :: I I II II II I I I I orf85-1 INSTSQTNTLNTEKSKLETYQAKLVSAQIALGSAEKKYKRQAALWKENATSKEDLESAQD 40 50 60 70 80 90 150 160 170 180 190 200 orf85ng ALAAAKANVAELKALIRQSKISINTAESDLGYTRITATMDGTWAIPVEEGQTVNAAQST I : I I I I II I I I I I I I I I I I II I I II I I I : I II I I II I I I I I I II II I I I II II I I I I I I orf85-1 AFAAAKANVAELKALIRQSKISINTAESELGYTRITATMDGTWAILVEEGQTVNAAQST 100 110 120 130 140 150 210 220 230 240 250 260 orf85ng PTIVQLANLDMMLNKMQIAEGDITKVKAGQDISFTILSEPDTPIKAKLDSVDPGLTTMSS II I I I I I I I I I I I I I II I II I II I I I II I II I II I I II I I I I I II I I I I I I I II I I I II I orf85-1 PTIVQLANLDMMLNKMQIAEGDITKVKAGQDISFTILSEPDTPIKAKLDSVDPGLTTMSS 160 170 180 190 200 210 270 280 290 300 310 320 orf85ng GGYNSSTDTASNAVYYYARSFVPNPDGKLATGMTTQNTVEIDGVKNVLLIPSLTVKNRGG II I II II I II II I II I I I I II I I II I I II I II I I I II II I I I I I I I I I : I I II I II II I I orf85-1 GGYNSSTDTASNAVYYYARSFVPNPDGKLATGMTTQNTVEIDGVKNVLIIPSLTVKNRGG 220 230 240 250 260 270 330 340 350 360 370 380 orf85ng KAFVRVLGADGKAVEREIRTGMKDSMNTEVKSGLKEGDKWISEITAAEQQESGERALGG II I II I II II I MM II I I I II : II I I I I II I I I I I I I II I II I I II I I II I I I I I I I I I orf85-1 KAFVRVLGADGKAAEREIRTGMRDSMNTEVKSGLKEGDKWISEITAAEQQESGERALGG 280 290 300 310 320 330 390 orf85ng PPRRX II II I orf85-l PPRRX In addition, ORF85ng shows significant homology to an E.coli membrane fusion protein: gi I 1787104 (AE000189) o380; 27% identical (27 gaps) to 332 residues from membrane fusion protein precursor, MTRC_NEIGO SW: P43505 (412 aa) [Escherichia coli] Length = 380 Score = 193 bits (485), Expect = 2e-48 Identities = 120/345 (34%), Positives = 182/345 (51%), Gaps = 13/345 (3%) Query: 29 PQAAYITETVRRGDISRTVSATGEISPSNLVSVGAQASGQIKKLYVKLGQQVKKGDLIAE 88 P Y T VR GD+ ++V ATG++ V VGAQ SGQ+K L V +G +VKK L+ Sbjct: 41 PVPTYQTLIVRPGDLQQSVLATGKLDALRKVDVGAQVSGQLKTLSVAIGDKVKKDQLLGV 100 Query: 89 INSTTQTNTIDMEKSKLETYQAKLVSAQIALGSAEKKYKRQAALWKDDATSKEXXXXXXX 148 1+ N I ++ L +A+ A+ L A Y RQ L + A S++ Sbjct: 101 IDPEQAENQIKEVEATLMELRAQRQQAEAELKLARVTYSRQQRLAQTKAVSQQDLDTAAT 160 Query: 149 XXXXXXXXXXXXXXXIRQSKISINTAESDLGYTRITATMDGTWAIPVEEGQTVNAAQST 208 I++++ S++TA+++L YTRI A M G V I +GQTV AAQ Sbjct: 161 EMAVKQAQIGTIDAQIKRNQASLDTAKTNLDYTRIVAPMAGEVTQITTLQGQTVIAAQQA 220 Query: 209 PTIVQLANLDMMLNKMQIAEGDITKVKAGQDISFTILSEPDTPIKAKLDSVDPGLTTMSS 268 P 1+ LA++ ML K Q++E D+ +K GQ FT+L +P T + ++ V P Sbjct: 221 PNILTLADMSAMLVKAQVSEADVIHLKPGQKAWFTVLGDPLTRYEGQIKDVLP 273 Query: 269 GGYNSSTDTASNAVYYYARSFVPNPDGKLATGMTTQNTVEIDGVKNVLLIPSLTVKNRGG 328 + + ++A++YYAR VPNP+G L MT Q +++ VKNVL IP + + G Sbjct: 274 TPEKVNDAIFYYARFEVPNPNGLLRLDMTAQVHIQLTDVKNVLTIPLSALGDPVG 328 Query: 329 KAFVRV-LGADGKAVEREIRTGMKDSMNTEVKSGLKEGDKWISE 372 +V L +G+ ERE+ G ++ + E+ GL+ GD+VVI E Sbjct: 329 DNRYKVKLLRNGETREREVTIGARNDTDVEIVKGLEAGDEWIGE 373 Based on this analysis, it was predicted that the proteins from Nmeningitidis and N.gonorrhoeae, and their epitopes, could be useful antigens for vaccines or diagnostics, or for raising antibodies. ORF85-1 (40.4kDa) was cloned in the pGex vectors and expressed in E.coli, as described above. The products of protein expression and purification were analyzed by SDS-PAGE. Figure 19A shows the results of affinity purification of the GST-fusion protein. Purified GST-fusion protein was used to immunise mice, whose sera were used for Western blot (Figure 19B), FACS analysis (Figure 19C), and ELISA (positive result). These experiments confirm that ORF85-1 is a surface-exposed protein, and that it is a useful immunogen. Example 92 The following partial DNA sequence was identified inN meningitidis <SEQ LD 773>: 1 ..ATTCCCGCCA CGATGACATT TGAACGCAGC GGCAATGCTT ACAAAATCGT 51 TTCGACGATT AAAGTGCCGC TATACAATAT CCGTTTCGAG TCCGGCGGTA 101 CGGTTGTCGG CAATACCCTG CACCCTACCT ACTATAGAGA CATACGCAGG 151 GGCAAACTGT ATGCGGAAgc CAAATTCGCC GACgGcAGCG TAACTTACGG 201 CAAAGCGGGC GAGAGCAAAA CCGAGCAAAG CCCCAAGGCT ATGGATTTGT 251 TCACGCTTGC CTGGCAGTTG GCGGCAAATG ACGCGAAACT CCCCCCGGGG 301 CTGAAAATCA CCAACGGCAA AAAACTTTAT TCCGTCGGCG GTTTGAATAA 351 GGCGGGTACA GGAAAATACA GCATAGGCGG CGTGGAAACC GAAGTCGTCA 401 AATATCGGGT GCGGCGCGGC GACGATGCGG TAATGTATTT cTTCGCACCG 451 TCCCTGAACA ATATTCCGGC ACAAATCGGC TATACCGACG ACGGCAAAAC 501 CTATACGCTG AAACTCAAAT CGGTGCAGAT CAACGGCCAG GCAGCCAAAC 551 CGTAA This coπesponds to the .amino acid sequence <SEQ ID 774; ORF120>: 1 .. IPΛΪMTFERS GNAYKIVSTI KVPLYNIRFE SGGTWGNTL HPTYYRDIRR 51 GKLYAEAKFA DGSVTYGKAG ESKTEQSPKA MDLFTLAWQL AANDAKLPPG 101 LKITNGKKLY SVGGLNKAGT GKYSIGGVET EWKYRVRRG DDAVMYFFAP 151 SLNNIPAQIG YTDDGKTYTL KLKSVQINGQ AAKP* Further work revealed the complete nucleotide sequence <SEQ ID 775>: 1 ATGATGAAGA CTTTTAAAAA TATATTTTCC GCCGCCATTT TGTCCGCCGC 51 CCTGCCGTGC GCGTATGCGG CAGGGCTGCC CCAATCCGCC GTGCTGCACT 101 ATTCCGGCAG CTACGGCATT CCCGCCACGA TGACATTTGA ACGCAGCGGC 151 AATGCTTACA AAATCGTTTC GACGATTAAA GTGCCGCTAT ACAATATCCG 201 TTTCGAGTCC GGCGGTACGG TTGTCGGCAA TACCCTGCAC CCTACCTACT 251 ATAGAGACAT ACGCAGGGGC AAACTGTATG CGGAAGCCAA ATTCGCCGAC 301 GGCAGCGTAA CTTACGGCAA AGCGGGCGAG AGCAAAACCG AGCAAAGCCC 351 CAAGGCTATG GATTTGTTCA CGCTTGCCTG GCAGTTGGCG GCAAATGACG 401 CGAAACTCCC CCCGGGGCTG AAAATCACCA ACGGCAAAAA ACTTTATTCC 451 GTCGGCGGTT TGAATAAGGC GGGTACAGGA AAATACAGCA TAGGCGGCGT 501 GGAAACCGAA GTCGTCAAAT ATCGGGTGCG GCGCGGCGAC GATGCGGTAA 551 TGTATTTCTT CGCACCGTCC CTGAACAATA TTCCGGCACA AATCGGCTAT 601 ACCGACGACG GCAAAACCTA TACGCTGAAA CTCAAATCGG TGCAGATCAA 651 CGGCCAGGCA GCCAAACCGT AA This coπesponds to the amino acid sequence <SEQ ED 776; ORF120-1>: 1 MMKTFKNIFS AAILSAALPC AYAAGLPQSA VLHYSGSYGI PATMTFERSG 51 NAYKIVSTIK VPLYNIRFES GGTWGNTLH PTYYRDIRRG KLYAEAKFAD 101 GSVTYGKAGE SKTEQSPKAM DLFTLAWQLA ANDAKLPPGL KITNGKKLYS 151 VGGLNKAGTG KYSIGGVETE WKYRVRRGD DAVMYFFAPS LNNIPAQIGY 201 TDDGKTYTLK LKSVQINGQA AKP* Computer analysis of this amino acid sequence gave the following results: Homology with a predicted ORF from N. meningitidis (sfrain Al ORF120 shows 92.4% identity over a 184aa overlap with an ORF (ORF120a) from strain A of N. meningitidis: 10 20 30 orf120. pep IPATMTFERSGNAYKIVSTIKVPLYNIRFE MM : I I I I I I I I II I I II I I II orf120a SAAILSAALPCAYAAGLPXSAVLHYSGSYGIPATXXXXXXXNAXKIVSTIKVPLYNIRFE 10 20 30 40 50 60 40 50 60 70 80 90 orf120.pep SGGTWGNTLHPTYYRDIRRGKLYAEAKFADGSVTYGKAGESKTEQSPKAMDLFTLAWQL I II I I I I I I I II I I I I I II I I II I I II I II I II I II I I I : I II I II I II I II I I I orf120a SGGTWGNTLHPTYYRDIRRGKLYAEAKFADGSVTYGKAXXXXXXQSPKAMDLFTLAWQL 70 80 90 100 110 120 100 110 120 130 140 150 orf120. pep AANDAKLPPGLKITNGKKLYSVGGLNKAGTGKYSIGGVETEWKYRVRRGDDAVMYFFAP II I I I I I I I I II I I II I II I I II I II I I II II I I II I I I II I I I I I II I I I I II I I II I I orf120a AANDAKLPPGLKITNGKKLYSVGGLNKAGTGKYSIGGVETEWKYRVRRGDDAVMYFFAP 130 140 150 160 170 180 160 170 180 orf120.pep SLNNIPAQIGYTDDGKTYTLKLKSVQINGQAAKPX I I I I I II II I II II I I II I II I II I II I I II II I I orf120a SLNNIPAQIGYTDDGKTYTLKLKSVQINGQAAKPX 190 200 210 220 The complete length ORF 120a nucleotide sequence <SEQ ID 777> is: 1 ATGATGAAGA CTTTTAAAAA TATATTTTCC GCCGCCATTT TGTCCGCCGC 51 CCTGCCGTGC GCGTATGCGG CAGGGCTGCC CNAATCCGCC GTGCTGCACT 101 ATTCCGGCAG CTACGGCATT CCCGCCACNA NNANNTNNGN ACNNNGNGNC 151 AATGCTTNCA AAATCGTTTC GACGATTAAA GTGCCGCTAT ACAATATCCG 201 TTTCGAGTCC GGCGGTACGG TTGTCGGCAA TACCCTGCAC CCTACCTACT 251 ATAGAGACAT ACGCAGGGGC AAACTGTATG CGGAAGCCAA ATTCGCCGAC 301 GGCAGCGTAA CCTACGGCAA AGCGGNNNNN ANCNNNNNNG NGCAAAGCCC 351 CAAGGCTATG GATTTGTTCA CGCTTGCNTG GCAGTTGGCG GCAAATGACG 401 CGAAACTCCC CCCGGGGCTG AAAATCACCA ACGGCAAAAA ACTTTATTCC 451 GTCGGCGGTT TGAATAAGGC GGGTACAGGA AAATACAGCA TAGGCGGCGT 501 GGAAACCGAA GTCGTCAAAT ATCGGGTGCG GCGCGGCGAC GATGCGGTAA 551 TGTATTTCTT CGCACCGTCC CTGAACAATA TTCCGGCACA AATCGGCTAT 601 ACCGACGACG GCAAAACCTA TACGCTGAAA CTCAAATCGG TGCAGATCAA 651 CGGCCAGGCA GCCAAACCGT AA This encodes a protein having amino acid sequence <SEQ ID 778>: 1 MMKTFKNIFS AAILSAALPC AYAAGLPXSA VLHYSGSYGI PATXXXXXXX 51 NAXKIVSTIK VPLYNIRFES GGTWGNTLH PTYYRDIRRG KLYAEAKFAD 101 GSVTYGKAXX XXXXQSPKAM DLFTLAWQLA ANDAKLPPGL KITNGKKLYS 151 VGGLNKAGTG KYSIGGVETE WKYRVRRGD DAVMYFFAPS LNNIPAQIGY 201 TDDGKTYTLK LKSVQINGQA AKP* ORF120a and ORF120-1 show 93.3% identity in 223 aa overlap: 10 20 30 40 50 60 orf120a . pep MMKTFKNIFSAAILSAALPCAYAAGLPXSAVLHYSGSYGIPATXXXXXXXNAXKIVSTIK II I I I II I I I II I I II II II I II I I I I I I II I II I I I I I II I : II I I I I I II orf120-1 MMKTFKNIFSAAILSAALPCAYAAGLPQSAVLHYSGSYGIPATMTFERSGNAYKIVSTIK 10 20 30 40 50 60 70 80 90 100 110 120 orf120a . pep VPLYNIRFESGGTWGNTLHPTYYRDIRRGKLYAEAKFADGSVTYGKAXXXXXXQSPKAM I I I II I I I II I I I II II II I I I I I II II II I I I I I I I I I I I II II II I : I I II I I orf120-1 VPLYNIRFESGGTWGNTLHPTYYRDIRRGKLYAEAKFADGSVTYGKAGESKTEQSPKAM 70 80 90 100 110 120 130 140 150 160 170 180 orf120a . pep DLFTLAWQLAANDAKLPPGLKITNGKKLYSVGGLNKAGTGKYSIGGVETEWKYRVRRGD II I I I I I II I I I I I I I II I II I II I I I I II I I I I II II I I I I I I I I II I I I II II II I I I orf120-1 DLFTLAWQLAANDAKLPPGLKITNGKKLYSVGGLNKAGTGKYSIGGVETEWKYRVRRGD 130 140 150 160 170 180 190 200 210 220 orf120a.pep DAVMYFFAPSLNNIPAQIGYTDDGKTYTLKLKSVQINGQAAKPX I I I II I II I II I II II I I II I II I I I I I I I I II I I I II I I I I I I orf120-1 DAVMYFFAPSLNNIPAQIGYTDDGKTYTLKLKSVQINGQAAKPX 190 200 210 220 Homology with a predicted ORF from N.εonorrhoeae ORF120 shows 97.8% identity over 184 aa overlap with a predicted ORF (ORF120ng) from N.gonorrhoeae: orf 120 . pep IPATMTFERSGNAYKIVSTIKVPLYNIRFE 30 I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I orf l20ng SAAILSAALPCAYAARLPQSAVLHYSGSYGIPATMTFERSGNAYKIVSTIKVPLYNIRFE 69 orf 120 . pep SGGTWGNTLHPTYYRDIRRGKLYAEAKFADGSVTYGKAGESKTEQSPKAMDLFTLAWQL 90 I I I I I I I I I I I I M I : I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I orf 12 Ong SGGTWGNTLHPAYYKDIRRGKLYAEAKFADGSVTYGKAGESKTEQSPKAMDLFTLAWQL 129 orf120.pep AANDAKLPPGLKITNGKKLYSVGGLNKAGTGKYSIGGVETEWKYRVRRGDDAVMYFFAP 150 II I II I II II I II II II I I I II I I I I I I I II I I II I I II I II I I II I I I I I I : I I I I II orfl20ng AANDAKLPPGLKITNGKKLYSVGGLNKAGTGKYSIGGVETEWKYRVRRGDDTVTYFFAP 189 orf120. pep SLNNIPAQIGYTDDGKTYTLKLKSVQINGQAAKP 184 II I II I I I I I II I I I II I I I I I I II II I I II I I I orfl20ng ΞLNNIPAQIGYTDDGKTYTLKLKSVQINGQAAKP 223 The complete length ORF120ng nucleotide sequence <SEQ ID 779> is: 1 ATGATGAAGA CTTTTAAAAA TATATTTTCC GCCGCCATTT TGTCCGCCGC 51 CCTGCCGTGC GCGTATGCGG CAAGGCTACC CCAATCCGCC GTGCTGCACT 101 ATTCCGGCAG CTACGGCATT CCCGCCACGA TGACATTTGA ACGCAGCGGC 151 AATGCTTACA AAATCGTTTC GACGATTAAA GTGCCGCTAT ACAATATCCG 201 TTTCGAATCC GGCGGTACGG TTGTCGGCAA TACCCTGCAC CCTGCCTACT 251 ATAAAGACAT ACGCAGGGGC AAACTGTATG CGGAAGCCAA ATTCGCCGAC 301 GGCAGCGTAA CCTACGGCAA AGCGGGCGAG AGCAAAACCG AGCAAAGCCC 351 CAAGGCTATG GATTTGTTCA CGCTTGCCTG GCAGTTGGCG GCAAATGACG 401 CGAAACTCCC CCCGGGTCTG AAAATCACCA ACGGCAAAAA ACTTTATTCC 451 GTCGGCGGCC TGAATAAGGC GGGTACGGGA AAATACAGCA TaggCGGCGT 501 GGAAACCGAA GTCGTCAAAT ATCGGGTGCG GCGCGGCGAC GATACGGTAA 551 CGTATTTCTT CGCACCGTCC CTGAACAATA TTCCGGCACA AATCGGCTAT 601 ACCGACGACG GCAAAACCTA TACGCTGAAG CTCAAATCGG TGCAGATCAA 651 CGGACAGGCC GCCAAACCGT AA This encodes a protein having amino acid sequence <SEQ LD 780>: 1 MMKTFKNIFS AAILSAALPC AYAARLPQSA VLHYSGSYGI PATMTFERSG 51 NAYKIVSTIK VPLYNIRFES GGTWGNTLH PAYYKDIRRG KLYAEAKFAD 101 GSVTYGKAGE SKTEQSPKAM DLFTLAWQLA ANDAKLPPGL KITNGKKLYΞ 151 VGGLNKAGTG KYSIGGVETE WKYRVRRGD DTVTYFFAPS LNNIPAQIGY 201 TDDGKTYTLK LKSVQINGQA AKP* In comparison with ORF120-1, ORF120ng shows 97.8% identity in 223 aa overlap: 10 20 30 40 50 60 orf120-1.pep MMKTFKNIFSAAILSAALPCAYAAGLPQSAVLHYSGSYGIPATMTFERSGNAYKIVSTIK I I II II I II I I I I I I I I I I I I I I I I I I I I I II I I I I I I I II I II I II I I I II II II I I I orf12Ong MMKTFKNIFSAAILSAALPCAYAARLPQSAVLHYSGSYGIPATMTFERSGNAYKIVSTIK 10 20 30 40 50 60 70 80 90 100 110 120 orf120-1.pep VPLYNIRFESGGTWGNTLHPTYYRDIRRGKLYAEAKFADGSVTYGKAGESKTEQSPKAM I I I I II I II I II I II I I I I II : I IM I II II I I I I I I I I II I II II I I I II I I I I I I I I I orf12Ong VPLYNIRFESGGTWGNTLHPAYYKDIRRGKLYAEAKFADGSVTYGKAGESKTEQSPKAM 70 80 90 100 110 120 130 140 150 160 170 180 orf120-1.pep DLFTLAWQLAANDAKLPPGLKITNGKKLYSVGGLNKAGTGKYSIGGVETEWKYRVRRGD M I I II I I I I II I II I II I I I I II I I I I I II I I II I I I II I II I II I I II I I I I I I I II I orfl20ng DLFTLAWQLAANDAKLPPGLKITNGKKLYSVGGLNKAGTGKYSIGGVETEWKYRVRRGD 130 140 150 160 170 180 190 200 210 220 orf120-1.pep DAVMYFFAPSLNNIPAQIGYTDDGKTYTLKLKSVQINGQAAKPX I: I II I I I I II I II I I I I I I I II I I I I I I II I I I I I I I I I I I I orfl20ng DTVTYFFAPSLNNIPAQIGYTDDGKTYTLKLKSVQINGQAAKPX 190 200 210 220 This analysis, including the presence of a putative leader sequence in the gonococcal protein suggests that the proteins from N. meningitidis and N.gonorrhoeae, and their epitopes, could be useful antigens for vaccines or diagnostics, or for raising antibodies. Example 93 The following partial DNA sequence was identified in N meningitidis <SEQ LD 781>: 1 ATGTATCGGA GGAAAGGGCG GGGCATCAAG CCGTGGATGG GTGCCGGTGC 51 .GCGTTTGCC GCCTTGGTCT GGCTGGTTTT CGCGCTCGGC GATACTTTGA 101 CTCCGTTTGC GGTTGCGGCG GTGCTGGCGT ATGTATTGGA CCCTTTGGTC 151 GAATGGTTGC AGAAAAAGGG TTTGAACCGT GCATCCGCTT CGATGTCTGT 201 GATGGTGTTT TCCTTGATTT TGTTGTTGGC ATTATTGTTG ATTATCGTCC 251 CTATGCTGGT CGGGCAGTTC AACAATTTGG CATCGCGCCT GCCCCAATTA 301 ATCGGTTTTA TGCAGAACAC GCTGCTGCCG TGGTTGAAAA ATACAATCGG 351 CGGATATGTG GAAATCGATC AGGCATCTAT TATTGCGTGG CTTCAGGCGC 401 ATACGGGAGA GTTGAGCAAC GCGCTTAAGG CGTGGTTTCC CGTTTTGATG 451 AGGCAGGGCG GCAATATT .. This coπesponds to the .amino acid sequence <SEQ ID 782; ORF121 > • 1 MYRRKGRGIK PWMGAGXAFA ALVWLVFALG DTLTPFAVAA VLAYVLDPLV 51 EWLQKKGLNR ASASMSVMVF SLILLLALLL IIVPMLVGQF NNLASRLPQL 101 IGFMQNTLLP WLKNTIGGYV EIDQASIIAW LQAHTGELSN ALKAWFPVLM 151 RQGGNI.. Further work revealed the complete nucleotide sequence <SEQ ID 783>: 1 ATGTATCGGA GGAAAGGGCG GGGCATCAAG CCGTGGATGG GTGCCGGTGC 51 GGCGTTTGCC GCCTTGGTCT GGCTGGTTTT CGCGCTCGGC GATACTTTGA 101 CTCCGTTTGC GGTTGCGGCG GTGCTGGCGT ATGTATTGGA CCCTTTGGTC 151 GAATGGTTGC AGAAAAAGGG TTTGAACCGT GCATCCGCTT CGATGTCTGT 201 GATGGTGTTT TCCTTGATTT TGTTGTTGGC ATTATTGTTG ATTATCGTCC 251 CTATGCTGGT CGGGCAGTTC AACAATTTGG CATCGCGCCT GCCCCAATTA 301 ATCGGTTTTA TGCAGAACAC GCTGCTGCCG TGGTTGAAAA ATACAATCGG 351 CGGATATGTG GAAATCGATC AGGCATCTAT TATTGCGTGG CTTCAGGCGC 401 ATACGGGAGA GTTGAGCAAC GCGCTTAAGG CGTGGTTTCC CGTTTTGATG 451 AGGCAGGGCG GCAATATTGT CAGCAGTATC GGCAACCTGC TGCTGCTTCC 501 CTTGCTGCTT TACTATTTCC TGCTGGATTG GCAGCGGTGG TCGTGCGGCA 551 TTGCCAAACT GGTTCCGAgG CGTTTTGCCG GTGCTTATAC GCGCATTACA 601 GGCAATTTGA ACGAGGTATT GGGCGAATTT TTGCGCGGGC AGCTTCTGGT 651 AATGCTGATT ATGGGCTTGG TTTACGGTTT GGGATTGGTG CTGGTCGGGC 701 TGGATTCGGG GTTTGCCATC GGTATGCTTG CCGGTATTTT GGTGTTTGTC 751 CCTTATCTCG GGGCGTTTAC GGGATTGCTG CTTGCCACCG TCGCCGCCTT 801 GCTCCAGTTC GGTTCGTGGA ACGGCATCCT ATCGGTTTGG GCGGTTTTTG 851 CCGTAGGACA GTTTCTCGAA AGTTTTTTCA TTACGCCGAA AATCGTGGGA 901 GACCGTATCG GGCTGTCGCC GTTTTGGGTT ATCTTTTCGC TGATGGCGTT 951 CGGGCAGCTG ATGGGCTTTG TCGGAATGTT GGCGGGATTG CCTTTGGCCG 1001 CCGTAACCTT GGTCTTGCTT CGCGAGGGCG TGCAGAAATA TTTTGCCGGC 1051 AGTTTTTACC GGGGCAGGTA G This coπesponds to the amino acid sequence <SEQ LD 784; ORF121-l>: 1 MYRRKGRGIK PWMGAGAAFA ALVWLVFALG DTLTPFAVAA VLAYVLDPLV 51 EWLQKKGLNR ASASMSVMVF SLILLLALLL IIVPMLVGQF NNLASRLPQL 101 IGFMQNTLLP WLKNTIGGYV EIDQASIIAW LQAHTGELSN ALKAWFPVLM 151 RQGGNIVSSI GNLLLLPLLL YYFLLDWQRW SCGIAKLVPR RFAGAYTRIT 201 GNLNEVLGEF LRGQLLVMLI MGLVYGLGLV LVGLDSGFAI GMLAGILVFV 251 PYLGAFTGLL LATVAALLQF GSWNGILSVW AVFAVGQFLE SFFITPKIVG 301 DRIGLSPFWV IFSLMAFGQL MGFVGMLAGL PLAAVTLVLL REGVQKYFAG 351 SFYRGR* Computer analysis of this amino acid sequence gave the following results: Homology with a predicted ORF from N. meningitidis (strain Al ORF121 shows 98.7% identity over a 156aa overlap with an ORF (ORF121a) from strain A of N. meningitidis: 10 20 30 40 50 60 orf121. pep MYRRKGRGIKPWMGAGXAFAALVWLVFALGDTLTPFAVAAVLAYVLDPLVEWLQKKGLNR I II I I II I I II I I II I I I I I I I I I II I I II I I I I M I I I I I II I I I I I II I I I II I I I orf12la MYRRKGRGIKPWMDAGAAFAALVWLVFALGDTLTPFAVAAVLAYVLDPLVEWLQKKGLNR 10 20 30 40 50 60 70 80 90 100 110 120 orf121. pep ASASMSVMVFSLILLLALLLIIVPMLVGQFNNLASRLPQLIGFMQNTLLPWLKNTIGGYV ^Soil I I II II I II II I I I I I II II I I I II I I I II II I I I I II II II II I I I II I I I I I I I I I orf121a ASASMSVMVFSLILLLALLLIIVPMLVGQFNNLASRLPQLIGFMQNTLLPWLKNTIGGYV 70 80 90 100 110 120 130 140 150 orf121. pep EIDQASIIAWLQAHTGELSNALKAWFPVLMRQGGNI I I I II II I II II I I I I II I I I I I I II I II I II II I I orf121a EIDQASIIAWLQAHTGELSNALKAWFPVLMRQGGNIVSSIGNLLLLPLLLYYFLLDWQRW 130 140 150 160 170 180 orf12la SCGIAKLVPRRFAGAYTRITGNLNEVLGEFLRGQLLVMLIMGLVYGLGLVLVGLDSGFAI 190 200 210 220 230 240 The complete length ORF121 a nucleotide sequence <SEQ ID 785> is: 1 ATGTATCGGA GGAAAGGGCG GGGCATCAAG CCGTGGATGG ATGCCGGTGC 51 GGCGTTTGCC GCCTTGGTCT GGCTGGTTTT CGCGCTCGGC GATACTTTGA 101 CTCCGTTTGC GGTTGCGGCG GTGCTGGCGT ATGTATTGGA CCCTTTGGTC 151 GAATGGTTGC AGAAAAAGGG TTTGAACCGT GCATCCGCTT CGATGTCTGT 201 GATGGTGTTT TCCTTGATTT TGTTGTTGGC ATTATTGTTG ATTATTGTCC 251 CTATGCTGGT CGGGCAGTTC AACAATTTGG CATCGCGCCT GCCCCAATTA 301 ATCGGTTTTA TGCAGAACAC GCTGCTGCCG TGGTTGAAAA ATACAATCGG 351 CGGATATGTG GAAATCGATC AGGCATCTAT TATTGCGTGG CTTCAGGCGC 401 ATACGGGCGA GTTGAGCAAC GCGCTTAAGG CGTGGTTTCC CGTTTTGATG 451 AGGCAGGGCG GCAATATTGT CAGCAGTATC GGCAACCTGC TGCTGCTTCC 501 CTTGCTGCTT TACTATTTCC TGCTGGATTG GCAGCGGTGG TCGTGCGGCA 551 TTGCCAAACT GGTTCCGAGG CGTTTTGCCG GTGCTTATAC GCGCATTACA 601 GGCAATTTGA ACGAGGTATT GGGCGAATTT TTGCGCGGGC AGCTTCTGGT 651 GATGCTGATT ATGGGTTTGG TTTACGGCTT GGGGTTGGTG CTGGTCGGGC 701 TGGATTCGGG GTTTGCAATC GGTATGGTTG CCGGTATTTT GGTTTTTGTT 751 CCCTATTTGG GCGCGTTTAC AGGACTGCTG CTGGCAACCG TCGCCGCCTT 801 GCTCCAGTTC GGTTCGTGGA ACGGCATCTT GGCTGTTTGG GCGGTTTTTG 851 CCGTAGGACA GTTTCTCGAA AGTTTTTTCA TTACGCCGAA AATCGTGGGA 901 GACCGTATCG GCCTGTCGCC GTTTTGGGTT ATCTTTTCGC TGATGGCGTT 951 CGGGCAGCTG ATGGGCTTTG TCGGAATGTT GGCCGGATTG CCTTTGGCCG 1001 CCGTAACCTT GGTCTTGCTT CGCGAGGGCG TGCAGAAATA TTTTGCCGGC 1051 AGTTTTTACC GGGGCAGGTA G This encodes a protein having amino acid sequence <SEQ ID 786>: 1 MYRRKGRGIK PWMDAGAAFA ALVWLVFALG DTLTPFAVAA VLAYVLDPLV 51 EWLQKKGLNR ASASMSVMVF SLILLLALLL IIVPMLVGQF NNLASRLPQL 101 IGFMQNTLLP WLKNTIGGYV EIDQASIIAW LQAHTGELSN ALKAWFPVLM 151 RQGGNIVSSI GNLLLLPLLL YYFLLDWQRW SCGIAKLVPR RFAGAYTRIT 201 GNLNEVLGEF LRGQLLVMLI MGLVYGLGLV LVGLDSGFAI GMVAGILVFV 251 PYLGAFTGLL LATVAALLQF GSWNGILAVW AVFAVGQFLE SFFITPKIVG 301 DRIGLSPFWV IFSLMAFGQL MGFVGMLAGL PLAAVTLVLL REGVQKYFAG 351 SFYRGR* ORF121a and ORF121-1 show99.2% identity in 356 aa overlap: 10 20 30 40 50 60 orf121a .pep MYRRKGRGIKPWMDAGAAFAALVWLVFALGDTLTPFAVAAVLAYVLDPLVEWLQKKGLNR I I I I I I I I I I I I I I II I I II I II I I I I I I I I I I I II I I I I I I I II I II I II II I I II I I orf121-1 MYRRKGRGIKPWMGAGAAFAALVWLVFALGDTLTPFAVAAVLAYVLDPLVEWLQKKGLNR 10 20 30 40 50 60 70 80 90 100 110 120 orf121a . pep ASASMSVMVFSLILLLALLLIIVPMLVGQFNNLASRLPQLIGFMQNTLLPWLKNTIGGYV I I I I I I I I II II I I I II I I I II I II I I I I I II I II I I I I I I I I II I II I I I I I I I I I II I orf121-1 ASASMSVMVFSLILLLALLLIIVPMLVGQFNNLASRLPQLIGFMQNTLLPWLKNTIGGYV 70 80 90 100 110 120 130 140 150 160 170 180 orf121a . pep EIDQASIIAWLQAHTGELSNALKAWFPVLMRQGGNIVSSIGNLLLLPLLLYYFLLDWQRW I I I I I I I I I I II I II I II I I II I I II II II I I II I I I II II II I I I I I I II I I I I I I I II orf121-1 EIDQASIIAWLQAHTGELSNALKAWFPVLMRQGGNIVSSIGNLLLLPLLLYYFLLDWQRW 130 140 150 160 170 180 190 200 210 220 230 240 orf12la . pep SCGIAKLVPRRFAGAYTRITGNLNEVLGEFLRGQLLVMLIMGLVYGLGLVLVGLDSGFAI I II I II I II I I I I I II I I I II I I I I I I I I I II II I I II I I II II I I I I I II I I II II I II orf121-1 SCGIAKLVPRRFAGAYTRITGNLNEVLGEFLRGQLLVMLIMGLVYGLGLVLVGLDSGFAI 190 200 210 220 230 240 250 260 270 280 290 300 orf121a . pep GMVAGILVFVPYLGAFTGLLLATVAALLQFGSWNGILAVWAVFAVGQFLESFFITPKIVG I M I I I II I II I II I I I I I II I I I I I II I I II I I I I I : I I I II I I I II I I I II I I I I II I orf121-1 GMLAGILVFVPYLGAFTGLLLATVAALLQFGSWNGILSVWAVFAVGQFLESFFITPKIVG 250 260 270 280 290 300 310 320 330 340 350 orf121a . pep DRIGLSPFWVIFSLMAFGQLMGFVGMLAGLPLAAVTLVLLREGVQKYFAGSFYRGRX II I II I I I II I II I I I I I II I I I I I I II II I I I II I I I II I I I II I I I I I I I I II I I orf121-1 DRIGLSPFWVIFSLMAFGQLMGFVGMLAGLPLAAVTLVLLREGVQKYFAGSFYRGRX 310 320 330 340 350 Homology with a predicted ORF from N.sonorrhoeae ORF121 shows 97.4% identity over a 156 aa overlap with a predicted ORF (ORF121ng) from N.gonorrhoeae: orf121. pep MYRRKGRGIKPWMGAGXAFAALVWLVFALGDTLTPFAVAAVLAYVLDPLVEWLQKKGLNR 60 II I II I I I I I I I II I I II I I I I I I I: I I I II I II I I I I II I II II I I I II I II I I I I II orfl21ng MYRRKGRGIKPWMGAGAAFAALVWLVYALGDTLTPFAVAAVLAYVLDPLVEWLQKKGLNR 60 orf121. pep ASASMSVMVFSLILLLALLLIIVPMLVGQFNNLASRLPQLIGFMQNTLLPWLKNTIGGYV 120 II I II I I I I I I I II I II II I II I I I I I I I I I I I I II II II I II I I I II II I I I II I I I I I orfl21ng ASASMSVMVFSLILLLALLLIIVPMLVGQFNNLASRLPQLIGFMQNTLLPWLKNTIGGYV 120 orf121. pep EIDQASIIAWLQAHTGELSNALKAWFPVLMRQGGNI 156 II I II I I I I I : I I I I I I I I I II I I I I I I I I: II I I I orfl21ng EIDQASIIAWFQAHTGELSNALKAWFPVLMKQGGNIVSTIGNLLLPPLLLYYFLLDWHRW 180 An ORF121ng nucleotide sequence <SEQ LD 787> was predicted to encode a protein having amino acid sequence <SEQ LD 788>: 1 MYRRKGRGIK PWMGAGAAFA ALVWLVYALG DTLTPFAVAA VLAYVLDPLV 51 EWLQKKGLNR ASASMSVMVF SLILLLALLL IIVPMLVGQF NNLASRLPQL 101 IGFMQNTLLP WLKNTIGGYV EIDQASIIAW FQAHTGELSN ALKAWFPVLM 151 KQGGNIVSTI GNLLLPPLLL YYFLLDWHRW SCGIPKLVPR RFAGAYTRIT 201 GNLNKVWGKF LRGQLLGETE RGAWCRVGR ECWEGGGARS RPSDDGWPRW 251 GGG* Further work revealed the following gonoccocal DNA sequence <SEQ ID 789>: 1 ATGTATCGGA GAAAAGGACG GGGCATCAAG CCGTGGATGG GTGCCGGCGC 51 GGCGTTTGCC GCCTTGGTCT GGCTGGTTTA CGCGCTCGGC GATACTTTGA 101 CTCCGTTTGC GGTTGCGGCG GTGCTGGCGT ATGTGTTGGA CCCTTTGGTC 151 GAATGGTTGC AGAAAAAGGG TTTGAACCGT GCATCCGCTT CGATGTCTGT 201 GATGGTGTTT TCCTTGATTT TGTTGTTGGC ATTATTGTTG ATTATTGTCC 251 CTATGCTGGT CGGGCAGTTC AATAATTTGG CATCTCGCCT GCCCCAATTA 301 ATCGGTTTTA TGCAGAACAC GCTGCTGCCG TGGTTGAAAA ATACAATCGG 351 CGGATATGTG GAAATCGATC AGGCATCTAT TATTGCGTGG TTTCAGGCGC 401 ATACGGGCGA GTTGAGCAAC GCGCTTAAGG CGTGGTTTCC CGTTTTGATG 451 AAACAGGGCG GCAATATTGT CAGCAGTATC GGCAACCTGC TGCTGCCGCC 501 CTTGCTGCTT TACTATTTCC TGCTGGATTG GCAGCGGTGG TCGTGCGGCA 551 TCGCCAAACT GGTTCCGAGG CGTTTTGCCG GTGCTTATAC GCGCATTACG 601 GGTAATTTGA ACGAGGTATT GGGCGAATTT TTGCGCGGTC AGCTTCTGGT 651 GATGCTGATT ATGGGCTTGG TTTACGGTTT GGGATTGATG CTAGTCGGAC 701 TGGATTCGGG ATTTGCCATC GGTATGGTTG CCGGTATTTT GGTGTTTGTC 751 CCCTATTTGG GTGCGTTTAC GGGATTGCTG CTTGCCACTG TTGCAGCCTT 801 GCTCCAGTTC GGTTCGTGGA ACGGAATCTT GGCTGTTTGG GCGGTTTTTG 851 CCGTCGGTCA GTTTCTCGAA AGTTTTTTCA TTACGCCGAA AATTGTAGGA 901 GACCGTATCG GCCTGTCGCC GTTTTGGGTT ATCTTTTCGC TGATGGCGTT 951 CGGAGAGCTG ATGGGCTTTG TCGGAATGTT GGCCGGATTG CCTTTGGCCG 1001 CCGTAACCTT GGTCTTGCTT CGCGAGGGCG CGCAGAAATA TTTTGCCGGC 1051 AGTTTTTACC GGGGCAGGTA G This coπesponds to the .amino acid sequence <SEQ ID 790; ORF121ng-l>: 1 MYRRKGRGIK PWMGAGAAFA ALVWLVYALG DTLTPFAVAA VLAYVLDPLV 51 EWLQKKGLNR ASASMSVMVF SLILLLALLL IIVPMLVGQF NNLASRLPQL 101 IGFMQNTLLP WLKNTIGGYV EIDQASIIAW FQAHTGELSN ALKAWFPVLM 151 KQGGNIVSSI GNLLLPPLLL YYFLLDWQRW SCGIAKLVPR RFAGAYTRIT 201 GNLNEVLGEF LRGQLLVMLI MGLVYGLGLM LVGLDSGFAI GMVAGILVFV 251 PYLGAFTGLL LATVAALLQF GSWNGILAVW AVFAVGQFLE SFFITPKIVG 301 DRIGLSPFWV IFSLMAFGEL MGFVGMLAGL PLAAVTLVLL REGAQKYFAG 351 SFYRGR* ORF121ng-l and ORF121-1 show 97.5% identity in 356 aa overlap: 10 20 30 40 50 60 orf121-1. pep MYRRKGRGIKPWMGAGAAFAALVWLVFALGDTLTPFAVAAVLAYVLDPLVEWLQKKGLNR I II II I I I II I I I I I I I I I I I I I I I I : I I I II I I I II I I I I I II I I I I I I II I I I I I I II orf12lng-1 MYRRKGRGIKPWMGAGAAFAALVWLVYALGDTLTPFAVAAVLAYVLDPLVEWLQKKGLNR 10 20 30 40 50 60 70 80 90 100 110 120 orf121-1. pep ASASMSVMVFSLILLLALLLIIVPMLVGQFNNLASRLPQLIGFMQNTLLPWLKNTIGGYV II I I II I II I I I I II I I I II I II I II I II I I I II I I I I I I I I I I I I I I II II I I I I I I II orfl21ng-l ASASMSVMVFSLILLLALLLIIVPMLVGQFNNLASRLPQLIGFMQNTLLPWLKNTIGGYV 70 80 90 100 110 120 130 140 150 160 170 180 orf121-1. pep EIDQASIIAWLQAHTGELSNALKAWFPVLMRQGGNIVSSIGNLLLLPLLLYYFLLDWQRW I I I II I I I II M I I I II II I I I I II I I I II : II I I I II II I II II II I II I I I II II I I orfl21ng-l EIDQASIIAWFQAHTGELSNALKAWFPVLMKQGGNIVSSIGNLLLPPLLLYYFLLDWQRW 130 140 150 160 170 180 190 200 210 220 230 240 orf121-1.pep SCGIAKLVPRRFAGAYTRITGNLNEVLGEFLRGQLLVMLIMGLVYGLGLVLVGLDSGFAI II I I II I I I I I I I I I I II I I I I II I I I I I I I I I II I I I I II II II I I II : I I I II I I I I I orfl21ng-l SCGIAKLVPRRFAGAYTRITGNLNEVLGEFLRGQLLVMLIMGLVYGLGLMLVGLDSGFAI 190 200 210 220 230 240 250 260 270 280 290 300 orf121-1. pep GMLAGILVFVPYLGAFTGLLLATVAALLQFGSWNGILSVWAVFAVGQFLESFFITPKIVG I M I I I II I I II I II I I I II I I I I II I II I II I I I MM I I I I I I I I II I I I I II I I I I I orfl21ng-l GMVAGILVFVPYLGAFTGLLLATVAALLQFGSWNGILAVWAVFAVGQFLESFFITPKIVG 250 260 270 280 290 300 310 320 330 340 350 orf121-1. pep DRIGLSPFWVIFSLMAFGQLMGFVGMLAGLPLAAVTLVLLREGVQKYFAGSFYRGRX I I I I I I I I II I I I I I I I I : I I II I II I I I I II I I I II I I I I I 1:1 I I II II I I II I I orfl21ng-l DRIGLSPFWVIFSLMAFGELMGFVGMLAGLPLAAVTLVLLREGAQKYFAGSFYRGRX 310 320 330 340 350 In addition, ORF121ng-l shows homology to a permease from H.influenzae: sp|P43969|PERM_HAEIN PUTATIVE PERMEASE PERM HOMOLOG Length = 349 Score = 69.9 bits (168), Expect = 2e-ll Identities = 67/317 (21%), Positives = 120/317 (37%), Gaps = 7/317 (2%) Query: 26 VYALGDTLTPFAVAAVLAYVLDPLVEWL-QKKGLNRASASMSVMVFSXXXXXXXXXXXVP 84 +Y GD + P +A VL+Y+L+ + +L Q R A++ + VP Sbjct: 32 IYFFGDLIAPLLIALVLSYLLEIPINFLNQYLKCPRMLATILIFGSFIGLAAVFFLVLVP 91 Query: 85 MLVGQFNNLASRLPQLIGFMQNTLLPWLKNTIGGYVE-IDQASIIAWFQAHTGELSNALK 143 ML Q +L S LP + N WL N Y E ID + + + F + ++ + Sbjct: 92 MLWNQTISLLSDLPAMF NKSNEWLLNLPKNYPELIDYSMVDSIFNSVREKILGFGE 147 Query: 144 AWFPVLMKQGGNIVSSIGNXXXXXXXXXXXXXDWQRWSCGIAKLVPRRFAGAYTRITGNL 203 + + + N+VS D G+++ +P+ A+ R + Sbjct: 148 SAVKLSLASIMNLVSLGIYAFLVPLMMFFMLKDKSELLQGVSRFLPKNRNLAFXRWK-EM 206 Query: 204 NEVLGEFLRGQXXXXXXXXXXXXXXXXXXXXDSGFAIGMVAGILVFVPYXXXXXXXXXXX 263 + + ++ G+ + + G+ V VPY Sbjct: 207 QQQISNYIHGKLLEILIVTLITYIIFLIFGLNYPLLLAFAVGLSVLVPYIGAVIVTIPVA 266 Query: 264 XXXXXQFGSWNGILAVWAVFAVGQFLESFFITPKIVGDRIGLSPFWVIFSLMAFGELMGF 323 QFG + FAV Q L+ + P + + + L P +1 S++ FG L GF Sbjct: 267 LVALFQFGISPTFWYIIIAFAVSQLLDGNLLVPYLFSEAVNLHPLIIIISVLIFGGLWGF 326 Query: 324 VGMLAGLPLAAVTLVLL 340 G+ +PLA + ++ Sbjct: 327 WGVFFAIPLATLVKAVI 343 Based on this analysis, including the presence of a putative leader sequence and transmembrane domains in the two proteins, it is predicted that the proteins from N. meningitidis .and N.gonorrhoeae, and their epitopes, could be useful antigens for vaccines or diagnostics, or for raising antibodies. Example 94 The following partial DNA sequence was identified in N. meningitidis <SEQ ED 791>: 1 ..ACTGCTTTTT CGGCGGCGCT GCGCTTGAGT CCATCATGAC TCGTCATATT 51 TTTGTCCTTT GGGAAACCGT ATCAACAAAC AGCCGCCATC TTAACATTTT 101 TTTGCACGTC CTGCCCGCCG CGTTCAAATG CGTACCAGCA ATACCGCCGC 151 CTGCGCCTCT ATGCCTTCCA TCCGCCCGAG ATAGCCGAGT TTTTCGTTGG 201 TTTTGCCTTT GATGTTGACG CACGAAATGT CTATGCCCAA ATCGGCGGCG 251 ATGTTGGCAC GCATTTGCGG AATGTGCGGC GCGAGTGTGG GTTTCTGTGC 301 AATCACGGTC GTATCGACAT TGACCGCCTG CCAACCCTGC GCCTGAACGC 351 TTTGATACGC CGCACGCAAA AGGACGCGGC TGTCCGCATC TTTGAACTCT 401 GCGGCGGTGT CGGGGAAATG GCTGCCGATA TCGCCCAAAC CTGCCGCACC 451 GAGCAGCGCG TCGGTAACGG CGTGCAGCAG CGCATCGGCA TCGGAGTGTC 501 CGAGCAGCCC TTTTTCAAAT GGGATTTCAA CTCCGCCAAG TATCAG.. This coπesponds to the .amino acid sequence <SEQ ID 792; ORF122>: 1 ..TAFSAALRLS PSXLVIFLSF GKPYQQTAAI LTFFCTSCPP RSNAYQQYRR 51 LRLYAFHPPE IAEFFVGFAF DVDARNVYAQ IGGDVGTHLR NVRRECGFLC 101 NHGRIDIDRL PTLRLNALIR RTQKDAAVRI FELCGGVGEM AADIAQTCRT 151 EQRVGNGVQQ RIGIGVSEQP FFKWDFNSAK YQ.. Further work revealed the complete nucleotide sequence <SEQ ID 793>: 1 ATATCGTACT GGGCAAGCAG TTCGCCGGAT TTTTTGGAAG TAGATACCGC 51 GCCTTTGATT TTTTTGCCGC TCTTACCCAA GGCTTCGATG AAAAAGTTGA 101 TGGTCGAGCC GGTACCGATG CCGATATATT CATTTTCGGG TACGAATTCG 151 ACTGCTTTTT CGGCGGCGAT GCGCTTGAGT TCGTCTTGTG TCGTCATATT 201 TTTGTCCTTT GGGAAACCGT ATCAACAAAC AGCCGCCATC TTAACATTTT 251 TTTGCACGTC CTGCCCGCCG CGTTCAAATG CGTACCAGCA ATACCGCCGC 301 CTGCGCCTCT ATGCCTTCCA TCCGCCCGAG ATAGCCGAGT TTTTCGTTGG 351 TTTTGCCTTT GATGTTGACG CACGAAATGT CTATGCCCAA ATCGGCGGCG 401 ATGTTGGCAC GCATTTGCGG AATGTGCGGC GCGAGTTTGG GTTTCTGTGC 451 AATCACGGTC GTATCGACAT TGACCGCCTG CCAACCCTGC GCCTGAACGC 501 TTTGATACGC CGCACGCAAA AGGACGCGGC TGTCCGCATC TTTGAACTCT 551 GCGGCGGTGT CGGGGAAATG GCTGCCGATA TCGCCCAAAC CTGCCGCACC 601 GAGCAGCGCG TCGGTAACGG CGTGCAGCAG CGCATCGGCA TCGGAGTGTC 651 CGAGCAGCCC TTTTTCAAAT GGGATTTCAA CTCCGCCAAG TATCAGCTTT 701 CTGCCTTCGG TCAGTTGGTG GACATCGTAG CCCTGTCCGA TACGGATGTT 751 CGTCATCGTT TGTGTTCCTG A This coπesponds to the amino acid sequence <SEQ ID 794; ORF122-l>: 1 ISYWASSSPD FLEVDTAPLI FLPLLPKASM KKLMVEPVPM PIYSFSGTNS 51 TAFSAAMRLS SSCWIFLSF GKPYQQTAAI LTFFCTSCPP RSNAYQQYRR 101 LRLYAFHPPE IAEFFVGFAF DVDARNVYAQ IGGDVGTHLR NVRREFGFLC 151 NHGRIDIDRL PTLRLNALIR RTQKDAAVRI FELCGGVGEM AADIAQTCRT 201 EQRVGNGVQQ RIGIGVSEQP FFKWDFNSAK YQLSAFGQLV DIVALSDTDV 251 RHRLCS* Computer .analysis of this amino acid sequence gave the following results: Homology with a predicted ORF from N.meninεitidis (sfrain Al ORF122 shows 94.0% identity over a 182aa overlap with an ORF (ORF122a) from sfrain A of N meningitidis: 10 20 30 orf122.pep TAFSAALRLSPSXLVIFLSFGKPYQQTAAI I II II I : I I I I : II II I I I I II I I I II I orf122a FLPLLPKASMKKLMVEPVPMPMYSFSGTΝSTAFSAAMRLSSSCWIFLSFGKPYQQTAAI 30 40 50 60 70 80 40 50 60 70 80 90 orf122. pep LTFFCTSCPPRSΝAYQQYRRLRLYAFHPPEIAEFFVGFAFDVDARΝVYAQIGGDVGTHLR I I I I I I I II I II II II I I I I I II I I I I I : I II I I II I I I II II I I II I I II I I II I orf122a LTFFXTSCPPRSΝPYQQYRRLRLYAFHAPEITEFFVGFAFXVDARΝVYAQIGGDVGTHLR 90 100 110 120 130 140 100 110 120 130 140 150 orf122. pep ΝVRRECGFLCΝHGRIDIDRLPTLRLΝALIRRTQKDAAVRIFELCGGVGEMAADIAQTCRT Mill II I I I I II I I I I I II II I I I I I II II II I I I I II I I II II I I I I II I I I I I I I I orf122a ΝMRREFGFLCΝHGRIDIDRLPTLRLΝALIRRTQKDAAVRIFELCGGVGEMAADIAQTCRT 150 160 170 180 190 200 160 170 180 orf122. pep EQRVGΝGVQQRIGIGVSEQPFFKWDFΝSAKYQ I II II I II II I I I I I I I I I II II I I I I I I I II orf122a EQRVGΝGVQQRIGIGVSEQPFFKWDFΝSAKYQLSAFGQLVDIVALSDTDVRHRLCSX 210 220 230 240 250 The complete length ORF122a nucleotide sequence <SEQ ID 795> is: 1 ATATCATATT GGGCAAGCAG TTCACTGGAT TTTTTGGAAG TAGATACCGC 51 GCCTTTGATT TTTTTGCCGC TCTTACCCAA GGCTTCGATG AAAAAGTTGA 101 TGGTCGAACC GGTACCGATG CCGATGTATT CGTTTTCGGG TACGAATTCG 151 ACTGCΝTTTT CGGCGGCGAT GCGCTTGAGT TCGTCTTGTG TCGTCATATT 201 TTTGTCCTTT GGGAAACCGT ATCAACAAAC AGCCGCCATC TTAACATTTT 251 TTΝΝΝACGTC CTGCCCGCCG CGTTCAAATC CTTACCAGCA ATACCGCCGC 301 CTGCGACTCT ATGCCTTCCA TGCGCCCGAG ATAACCGAGT TTTTCGTTGG 351 TTTTGCCTTT GAΝGTTGACG CACGAAATGT CTATGCCCAA ATCGGCGGCG 401 ATGTTGGCAC GCATTTGCGG AATATGCGGC GCGAGTTTGG GTTTCTGTGC 451 AATCACGGTC GTATCGACAT TGACCGCCTG CCAACCCTGC GCCTGAACGC 501 TTTGATACGC CGCACGCAAA AGGACGCGGC TGTCCGCATC TTTGAACTCT 551 GCGGCGGTGT CGGGGAAATG GCTGCCGATA TCGCCCAAAC CTGCCGCACC 601 GAGCAGCGCG TCGGTAACGG CGTGCAGCAG CGCATCGGCA TCGGAGTGTC 651 CGAGCAGCCC TTTTTCAAAT GGGATTTCAA CTCCGCCAAG TATCAGCTTT 701 CTGCCTTCGG TCAGTTGGTG GACATCGTAG CCCTGTCCGA TACGGATGTT 751 CGTCATCGTT TGTGTTCCTG A This encodes a protein having amino acid sequence <SEQ ID 796>: 1 ISYWASSSLD FLEVDTAPLI FLPLLPKASM KKLMVEPVPM PMYSFSGTΝS 51 TAFSAAMRLS SSCWIFLSF GKPYQQTAAI LTFFXTSCPP RSΝPYQQYRR 101 LRLYAFHAPE ITEFFVGFAF XVDARΝVYAQ IGGDVGTHLR ΝMRREFGFLC 151 NHGRIDIDRL PTLRLNALIR RTQKDAAVRI FELCGGVGEM AADIAQTCRT 201 EQRVGNGVQQ RIGIGVSEQP FFKWDFNSAK YQLSAFGQLV DIVALSDTDV 251 RHRLCS* ORF122a and ORF122-1 show 96.9% identity in 256 aa overlap: 10 20 30 40 50 60 orf122a . pep ISYWASSSLDFLEVDTAPLIFLPLLPKASMKKLMVEPVPMPMYSFSGTNSTAFSAAMRLS I I I I I II I I I I II I I I II I II II I I II II I I I II II I I I I: I II II I I II I I I I I I II I orf122-1 ISYWASSSPDFLEVDTAPLIFLPLLPKASMKKLMVEPVPMPIYSFSGTNSTAFSAAMRLS 10 20 30 40 50 60 70 80 90 100 110 120 orf122a . pep SSCWIFLSFGKPYQQTAAILTFFXTSCPPRSNPYQQYRRLRLYAFHAPEITEFFVGFAF I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I M I I I I I I I orf 122-1 SSCWIFLSFGKPYQQTAAILTFFCTSCPPRSNAYQQYRRLRLYAFHPPEIAEFFVGFAF 70 30 90 100 110 120 130 140 150 160 170 180 orf122a. pep XVDARNVYAQIGGDVGTHLRNMRREFGFLCNHGRIDIDRLPTLRLNALIRRTQKDAAVRI I II II II I I I I I I I II II I I : I I II II II I I II I I I I II I II I I I I I I I I II I I I I I II orf122-1 DVDARNVYAQIGGDVGTHLRNVRREFGFLCNHGRIDIDRLPTLRLNALIRRTQKDAAVRI 130 140 150 160 170 180 190 200 210 220 230 240 orf122a . pep FELCGGVGEMAADIAQTCRTEQRVGNGVQQRIGIGVSEQPFFKWDFNSAKYQLSAFGQLV I I II II I II I I I I II I I I II I I I I I I I I II II I II II II I I I I II I I I II I I II II II I I orf122-1 FELCGGVGEMAADIAQTCRTEQRVGNGVQQRIGIGVSEQPFFKWDFNSAKYQLSAFGQLV 190 200 210 220 230 240 250 orf122a. pep DIVALSDTDVRHRLCSX I II I II I II I I I I II I I orfl22-l DIVALSDTDVRHRLCSX 250 Homology with a predicted ORF from N.εonorrhoeae ORF122 shows 89.6% identity over a 182 aa overlap with a predicted ORF (ORF122ng) from N.gonorrhoeae: orf122. pep TAFSAALRLSPSXLVIFLSFGKPYQQTAAI 30 I I II I I : II I I : I I I I I I I I I II I I I I I orfl22ng FLPLLPKASMKKLMVEPVPMPMYSFSGTNSTAFSAAMRLSSSCWIFLSFGKPYQQTAAI 30 orf122. pep LTFFCTSCPPRSNAYQQYRRLRLYAFHPPEIAEFFVGFAFDVDARNVYAQIGGDVGTHLR 90 I I I II I I I II I I I I I I II I I II I II I II II II I II I I I I : I I II : : I I II II I I I I I orfl22ng LTFFCTSWPPRSNPYQQYRRLRLYAFHPPEIAEFFVGFAFDIDARNIDTQIGGDVGTHLR 140 orf122. pep NVRRECGFLCNHGRIDIDRLPTLRLNALIRRTQKDAAVRIFELCGGVGEMAADIAQTCRT 150 III I I II I II I II I IM I I I I II I II I I I I I I I I I I II I I I II I II : I II I: II I II I orfl22ng NVRCEFGFLCNHGRIDIDHLPTLRLNALIRRTQKDAAVRIFELCGGVGKMAADVAQTCRT 200 orf122. pep EQRVGNGVQQRIGIGVSEQPFFKWDFNSAKYQ 182 I II I I II I I I M I I : II I I I I I I I II I II I orfl22ng EQRVGNGVQQRVGIRMPEQPFFKWDFNSAKYQLSAFGQLVDIVALSDTDIRHRLCS 256 The complete length ORF122ng nucleotide sequence <SEQ ID 797> is: 1 ATGTCGTACC GGGCAAGCAG TTCGCCGGAT TTTTTGGAGG TTGAAACCGC 51 GCCTTTGATT TTTTTACCGC TTTTGCCCAA GGCTTCGATG AAGAAATTGa 101 tgGTCGAACC GgtaCCGATG CCGATGTATT CGTTTTCGGG TACGAATTCG 151 ACTGCTTTTT CGGCGGCGAT GCGCttgAgt TCgtcttgcg TcgTCATATT 201 TTTAtccttt gGGAAaccct atcaAcaAAc agccgccatC TTAACATTTT 251 TTTGCACGtc ctggccgccg cgttcaAATc cgtaccaGca ataccgccgc 301 ctgcgcctCT AtgcCTTCCA TCCGCCCGAG ATAGCCGAGT TTTTCGTTGG 351 TTTTGCCTTT GATatTGACG CACGAAATAT CGatacCCAa atcggcgGCG 401 ATGTTGGCAC GCATTTGCGG AATGTGCGGT GCGAGTTTGG GTTTCTGTGC 451 AATCACGGTC GTATCGACAT TGACCACCTG CCAACCCTGC GCCTGAACGC 501 TTTGATACGC CGCACGCAAA AGGACGCGGC TGTCCGCATC TTTGAACTCT 551 GCGGCGGTGT CGGGAAAATG GCTGCCGATG TCGCCCAAAC CTGCCGCACC 601 GAGCAGCgcg tcggtaaCGG CGTGCAGCAG cgcgTcgGCA TCCGAATGCC 651 CGAGCAGCCC TTTTTCAAAT GGGATTTCAA CTCCGCCAAG TATCAGCTTT 701 CTGCCTTCGG TCAATTGGTG GACATCGTAG CCCTGTCCGA TACGGATATT 751 CGTCATCGTT TGTGTTCCTG A This encodes a protem having amino acid sequence <SEQ LD 798>: 1 MSYRASSSPD FLEVETAPLI FLPLLPKASM KKLMVEPVPM PMYSFSGTNS 51 TAFSAAMRLS SSCWIFLSF GKPYQQTAAI LTFFCTSWPP RSNPYQQYRR 101 LRLYAFHPPE IAEFFVGFAF DIDARNIDTQ IGGDVGTHLR NVRCEFGFLC 151 NHGRIDIDHL PTLRLNALIR RTQKDAAVRI FELCGGVGKM AADVAQTCRT 201 EQRVGNGVQQ RVGIRMPEQP FFKWDFNSAK YQLSAFGQLV DIVALSDTDI 251 RHRLCS* ORF122ng and ORF122-1 show 92.6% identity in 256 aa overlap: 10 20 30 40 50 60 orf122-1. pep ISYWASSSPDFLEVDTAPLIFLPLLPKASMKKLMVEPVPMPIYSFSGTNSTAFSAAMRLS Ml I I I I I I I II I : II II I I I I II I I I II II I II I I II I I : I II I II I I II I I I I I I I I orfl22ng MSYRASSSPDFLEVETAPLIFLPLLPKASMKKLMVEPVPMPMYSFSGTNSTAFSAAMRLS 10 20 30 40 50 60 70 80 90 100 110 120 orfl22-l.pep SSCWIFLSFGKPYQQTAAILTFFCTSCPPRSNAYQQYRRLRLYAFHPPEIAEFFVGFAF I I I I II I I I I I I I I II I I II I I I I I II I I I I I I I I I I I II II I I I II I II I I I I I I I I orfl22ng SSCWIFLSFGKPYQQTAAILTFFCTSWPPRSNPYQQYRRLRLYAFHPPEIAEFFVGFAF 70 80 90 100 110 120 130 140 150 160 170 180 orf122-1. pep DVDARNVYAQIGGDVGTHLRNVRREFGFLCNHGRIDIDRLPTLRLNALIRRTQKDAAVRI MMM: : I M II II I I II I II I II I I I I I II I I I I : I II I I I I II I I II I I I I I I II orfl22ng DIDARNIDTQIGGDVGTHLRNVRCEFGFLCNHGRIDIDHLPTLRLNALIRRTQKDAAVRI 130 140 150 160 170 180 190 200 210 220 230 240 orf122-1. pep FELCGGVGEMAADIAQTCRTEQRVGNGVQQRIGIGVSEQPFFKWDFNSAKYQLSAFGQLV I I I II II I : I I II : I I I I I I II I I I I II I II : II : I I I II II I I I II I II I I I I I I II orfl22ng FELCGGVGKMAADVAQTCRTEQRVGNGVQQRVGIRMPEQPFFKWDFNSAKYQLSAFGQLV 190 200 210 220 230 240 250 orf122-1. pep DIVALSDTDVRHRLCSX II II I I II I : II I I II I orfl22ng DIVALSDTDIRHRLCSX 250 Based on this analysis, it is predicted that the proteins from N. meningitidis and N.gonorrhoeae, and their epitopes, could be useful antigens for vaccines or diagnostics, or for raising antibodies. Example 95 The following partial DNA sequence was identified in N. meningitidis <SEQ ID 799>: 1 .. GCCGGCGCGA GTGCGAACAA CATTTCCGCG CGTTTTGCGG AAACACCCGT 51 CGCTGTCAGC GTTACCCTGA TCGGCACGGT ACTTGCCGTC ATGCTGCCCG 101 TTACCGAATA TGAAAACTTC CTGCTGCTTA TCGGCTCGGT ATTTGCGCCG 151 ATGgGGCGGA JJTTTGATTGC CGACTTTTTC GTCTTGAAAC GGCGTGA This coπesponds to the .amino acid sequence <SEQ ID 800; ORF125>: 1 ..AGASANNISA RFAETPVAVS VTLIGTVLAV MLPVTEYENF LLLIGSVFAP 51 MGGFDCRLFR LETA* Further work revealed the complete nucleotide sequence <SEQ ED 801>: 1 ATGTCGGGCA ATGCCTCCTC TCCTTCATCT TCCTCCGCCA TCGGGCTGAT 51 TTGGTTCGGC GCGGCGGTAT CGATTGCCGA AATCAGCACG GGTACGCTGC 101 TTGCGCCTTT GGGCTGGCAG CGCGGTCTGG CGGCTCTACT TTTGGGTCAT 151 GCCGTCGGCG GCGCGCTGTT TTTTGCGGCG GCGTATATCG GCGCACTGAC 201 CGGACGCAGC TCGATGGAAA GCGTGCGCCT GTCGTTCGGC AAACGCGGTT 251 CAGTGCTGTT TTCCGTGGCG AATATGCTGC AACTGGCCGG CTGGACGGCG 301 GTGATGATTT ACGCCGGCGC AACGGTCAGC TCCGCTTTGG GCAAAGTGTT 351 GTGGGACGGC GAATCTTTTG TCTGGTGGGC ATTGGCAAAC GGCGCGCTGA 401 TTGTGCTGTG GCTGGTTTTC GGCGCACGCA AAACAGGCGG GCTGAAAACC 451 GTTTCGATGC TGCTGATGCT GTTGGCGGTT CTGTGGCTGA GTGCCGAAGT 501 CTTTTCCACG GCAGGCAGCA CCGCCGCACA GGTTTCAGAC GGCATGAGTT 551 TCGGAACGGC AGTCGAGCTG TCCGCCGTGA TGCCGCTTTC CTGGCTGCCG 601 CTTGCCGCCG ACTACACGCG CCACGCGCGC CGCCCGTTTG CGGCAACCCT 651 GACGGCAACG CTCGCCTACA CGCTGACCGG CTGCTGGATG TATGCCTTGG 701 GTTTGGCAGC GGCGTTGTTC ACCGGAGAAA CCGACGTGGC AAAAATCCTG 751 CTGGGCGCAG GTTTGGGTGC GGCAGGCATT TTGGCGGTCG TCCTCTCCAC 801 CGTTACCACA ACGTTTCTCG ATGCCTATTC CGCCGGCGCG AGTGCGAACA 851 ACATTTCCGC GCGTTTTGCG GAAACACCCG TCGCTGTCGG CGTTACCCTG 901 ATCGGCACGG TACTTGCCGT CATGCTGCCC GTTACCGAAT ATGAAAACTT 951 CCTGCTGCTT ATCGGCTCGG TATTTGCGCC GATGGCGGCG GTTTTGATTG 1001 CCGACTTTTT CGTCTTGAAA CGGCGTGAGG AGATTGAAGG CTTTGACTTT 1051 GCCGGACTGG TTCTGTGGCT TGCGGGCTTC ATCCTCTACC GCTTCCTGCT 1101 CTCGTCCGGC TGGGAAAGCA GCATCGGTCT GACCGCCCCC GTAATGTCTG 1151 CCGTTGCCAT TGCCACCGTA TCGGTACGCC TTTTCTTTAA AAAAACCCAA 1201 TCTTTACAAA GGAACCCGTC ATGA This coπesponds to the amino acid sequence <SEQ ID 802; ORF125-l>: 1 MSGNASSPSS SSAIGLIWFG AAVSIAEIST GTLLAPLGWQ RGLAALLLGH 51 AVGGALFFAA AYIGALTGRS SMESVRLSFG KRGSVLFSVA NMLQLAGWTA 101 VMIYAGATVS SALGKVLWDG ESFVWWALAN GALIVLWLVF GARKTGGLKT 151 VSMLLMLLAV LWLSAEVFST AGSTAAQVSD GMSFGTAVEL SAVMPLSWLP 201 LAADYTRHAR RPFAATLTAT LAYTLTGCWM YALGLAAALF TGETDVAKIL 251 LGAGLGAAGI LAWLSTVTT TFLDAYSAGA SANNISARFA ETPVAVGVTL 301 IGTVLAVMLP VTEYENFLLL IGSVFAPMAA VLIADFFVLK RREEIEGFDF 351 AGLVLWLAGF ILYRFLLSSG WESSIGLTAP VMSAVAIATV SVRLFFKKTQ 401 SLQRNPS* Computer analysis of this amino acid sequence gave the following results: Homology with a predicted ORF from N.meninsitidis (sfrain A) ORF125 shows 76.5% identity over a 5 laa overlap with an ORF (ORF125a) from strain A of N. meningitidis: 10 20 30 orf 125 . pep AGASANNISARFAETPVAVSVTLIGTVLAV I I : I I I I I I I : : : I I : I I : I : : : I I : I I I orf 125a KILLGAGLGAAGILAWLSTVTTTFLDAYSAGVSANNISAKLSEIPIAVAVAWGTLLAV 250 260 270 280 290 300 40 50 60 orf125.pep MLPVTEYENFLLLIGSVFAPMGGFDCRLFRLETAX : I I I II I I II I II I II II I II: orf125a LLPVTEYENFLLLIGSVFAPMAAVLIADFFVLKRREEIEG 310 320 330 340 The ORF125a partial nucleotide sequence <SEQ ID 803> is: 1 ATGTCGGGCA ATGCCTCCTC TCNTTCATCT TCCGCCGCCA TCGGGCTGAT 51 TTGGTTCGGC GCGGCGGTAT CGATTGCCGA AATCAGCACG GGTACACTGC 101 TTGCGCCTTT GGGCTGGCAG CGCGGTCTGG CNGCTCTGCT TTTGGGTCAT 151 GCCGTCGGCG GCGCGCTGTT TTTTGCGGCG GCGTATATCG GCGCACTGAC 201 CGGACNCANC TCGATGGAAA GCGTGCGCCT GTCGTTCGGC AAACGCGGTT 251 CAGTGCTGTT TTCCGTGGCG AATATGCTGC AACTGGCCGG CTGGACGGCG 301 GTGATGATTT ACGCCGGCGC AACGGTCAGC TCCGCTTTGG GCAAAGTGTT 351 GTGGGACGGC GAATCTTTTG TCTGGTGGGC ATTGGCAAAC GGCGCGCTGA 401 TTGTGCTGTG GCTGGTTTTC GGCGCACGCA AAACAGGCGG GCTGAAAACC 451 GTTTCGATGC TGCTGATGCT GTTGGCGGTT CTGTGGCTGA GTGCCGAANT 501 NTTTTCCACG GCAGGCAGCA CCGCCGCANN GGTNNCAGAC GGCATGAGTT 551 TCGGAACGGC AGTCGAGCTG TCCGCCGTNA TGCCGCTTTC TTGGCTGCCG 601 CTGGCCGCCG ACTACACGCG CCACGCGCGC CGCCCGTTTG CGGCAACCCT 651 GACGGCAACG CTCGCCTACA CGCTGACCGG CTGCTGGATG TATGCCTTGG 701 GTTTGGCAGC GGCGTTGTTC ACCGGAGAAA CCGACGTGGC AAAAATCCTG 751 CTGGGCGCAG GTTTGGGTGC GGCAGGCATT TTGGCGGTCG TCCTGTCGAC 801 CGTTACCACC ACTTTTCTCG ATGCNTACTC CGCCGGCGTA AGTGCCAACA 851 ATATTTCCGC CAAACTTTCG GAAATACCNA TCGCCGTTGC CGTCGCCGTT 901 GTCGGCACAC TGCTTGCCGT CCTCCTGCCC GTTACCGAAT ATGAAAACTT 951 CCTGCTGCTT ATCGGCTCGG TATTTGCGCC GATGGCGGCG GTTTTGATTG 1001 CCGACTTTTT CGTCTTGAAA CGGCGTGAGG AGATTGAAGG C.. This encodes a protein having the partial amino acid sequence <SEQ ED 804>: 1 MSGNASSXSS SAAIGLIWFG AAVSIAEIST GTLLAPLGWQ RGLAALLLGH 51 AVGGALFFAA AYIGALTGXX SMESVRLSFG KRGSVLFSVA NMLQLAGWTA 101 VMIYAGATVS SALGKVLWDG ESFVWWALAN GALIVLWLVF GARKTGGLKT 151 VSMLLMLLAV LWLSAEXFST AGSTAAXVXD GMSFGTAVEL SAVMPLSWLP 201 LAADYTRHAR RPFAATLTAT LAYTLTGCWM YALGLAAALF TGETDVAKIL 251 LGAGLGAAGI LAWLSTVTT TFLDAYSAGV SANNISAKLS EIPIAVAVAV 301 VGTLLAVLLP VTEYENFLLL IGSVFAPMAA VLIADFFVLK RREEIEG.. ORF125a and ORF125-1 show 94.5% identity in 347 aa overlap: 10 20 30 40 50 60 orf125a . pep MSGNASSXSSSAAIGLIWFGAAVSIAEISTGTLLAPLGWQRGLAALLLGHAVGGALFFAA I I I I I I I I I I : I I I I II I II II I I II I I I I I I II I I I I I I I II II I I I I II I I II II II orf125-1 MSGNASSPSSSSAIGLIWFGAAVSIAEISTGTLLAPLGWQRGLAALLLGHAVGGALFFAA 10 20 30 40 50 60 70 80 90 100 110 120 orf125a . pep AYIGALTGXXSMESVRLSFGKRGSVLFSVANMLQLAGWTAVMIYAGATVSSALGKVLWDG I I I II I I I II I I I I II I I I I II I II I II I I I I II II I I I I I II I I I II I I II I I I I I I orf125-1 AYIGALTGRSSMESVRLSFGKRGSVLFSVANMLQLAGWTAVMIYAGATVSSALGKVLWDG 70 80 90 100 110 120 130 140 150 160 170 180 orf125a . pep ESFVWWALANGALIVLWLVFGARKTGGLKTVSMLLMLLAVLWLSAEXFSTAGSTAAXVXD I I II II I I I II II I II I I I I I II I I II I I I II I II I II I I I I I II I I I I I I I II I I I orf125-1 ESFVWWALANGALIVLWLVFGARKTGGLKTVSMLLMLLAVLWLSAEVFSTAGSTAAQVSD 130 140 150 160 170 180 190 200 210 220 230 240 orf125a . pep GMSFGTAVELSAVMPLSWLPLAADYTRHARRPFAATLTATLAYTLTGCWMYALGLAAALF I I I I II I I I II II I II I I I I I II I I II I II I I I I II I I II I I II I I I II I I I I I I II I I I orf125-1 GMSFGTAVELSAVMPLSWLPLAADYTRHARRPFAATLTATLAYTLTGCWMYALGLAAALF 190 200 210 220 230 240 250 260 270 280 290 300 orf125a . pep TGETDVAKILLGAGLGAAGILAWLSTVTTTFLDAYSAGVSANNISAKLSEIPIAVAVAV I II I II I I I II II I II II I I I I II I II I II I I II I II II : II II II I ::: I I : I I : I :: orf125-1 TGETDVAKILLGAGLGAAGILAWLSTVTTTFLDAYSAGASANNISARFAETPVAVGVTL 250 260 270 280 290 300 310 320 330 340 orf125a . pep VGTLLAVLLPVTEYENFLLLIGSVFAPMAAVLIADFFVLKRREEIEG : II : II I : I I I II I II I I I I I I II I II I I I I II I I I I I I II II I I I I orf125-1 IGTVLAVMLPVTEYENFLLLIGSVFAPMAAVLIADFFVLKRREEIEGFDFAGLVLWLAGF 310 320 330 340 350 360 Homology with a predicted ORF from N.εonorrhoeae ORF125 shows 86.2% identity over a 65aa overlap with a predicted ORF (ORF125ng) from N.gonorrhoeae: orf125.pep AGASANNISARFAETPVAVSVTLIGTVLAV 30 I II I I I I I I II II I I I I I : I I II I I I I I orfl25ng KILLGAGLGITGILAWLSTVTTTFLDTYSAGASANNISARFAEIPVAVGVTLIRTVLAV 308 orf125.pep MLPVTEYENFLLLIGSVFAPM-GGFDCRLFRLETA 64 I I I I I I I : I I I II I MMM I II I II I I MM orfl25ng MLPVTEYKNFLLLIRSVFGPMAGGFDCRLFCLKTA 343 An ORF125ng nucleotide sequence <SEQ LD 805> was pr-edicted to encode a protein having amino acid sequence <SEQ ID 806>: 1 MSGNASSPSS SAAIGLVWFG AAVSIAEIST GTLLAPLGWQ RGLAALLLGH 51 AVGGALFFAA AYIGALTGRS SMESVRLSFG KCGSVLFSVA NMLQLAGWTA 101 VMIYVGATVS SALGKVLWDG ESFVWWALAN GALIVLWLVF GARRTGGLKT 151 VSMLLMLLAV LWLSVEVFAS SGTNAAPAVS DGMTFGTAVE LSAVMPLSWL 201 PLAADYTRQA RRPFAATLTA TLAYTLTGCW MYALGLAAAL FTGETDVAKI_ 251 LLGAGLGITG ILAWLSTVT TTFLDTYSAG ASANNISARF AEIPVAVGVT 301 LIRTVLAVML PVTEYKNFLL LIRSVFGPMA GGFDCRLFCL KTA* Further work revealed the following gonococcal DNA sequence <SEQ ID 807>: 1 ATGTCGGGCA ATGCCTCCTC TCCTTCATCT TCCGCCGCCA TCGGGCTGGT 51 TTGGTTCGGC GCGGCGGTAT CGATTGCCGA AATCAGCACG GGTACGCTGC 101 TCGCCCCCTT GGGCTGGCAG CGCGGTCTGG CGGCCCTGCT TTTGGGTCAT 151 GCCGTCGGCG GCGCGCTGTT TTTTGCGGCG GCGTATATCG GCGCACTGAC 201 CGGACGCAGC TCGATGGAAA GTGTGCGCCT GTCGTTCGGC AAATGCGGTT 251 CAGTGCTGTT TTCCGTGGCG AATATGCTGC AACTGGCCGG CTGGACGGCG 301 GTGATGATTT ACGTCGGCGC AACGGTCAGC TCCGCTTTGG GCAAAGTGTT 351 GTGGGACGGC GAATCCTTTG TCTGGTGGGC ATTGGCAAAC GGCGCACTGA 401 TCGTGCTGTG GCTGGTTTTC GGCGCACGCA GAACGGGCGG GCTGAAAACC 451 GTTTCGATGC TGCTGATGCT GCTTGCCGTG TTGTGGTTGA GCGTCGAAGT 501 GTTCGCTTCG TCCGGCACAA ACGCCGCGCC CGCCGTTTCA GACGGCATGA 551 CCTTCGGAAC GGCAGTCGAA CTGTCCGCCG TCATGCCGCT TTCCTGGCTG 601 CCGCTGGCCG CCGACTACAC GCGCCAAGCA CGCCGCCCGT TTGCGGCAAC 651 CCTGACGGCA ACGCTCGCCT ATACGCTGAC GGGCTGCTGG ATGTATGCCT 701 TGGGTTTGGC GGCGGCTCTG TTTACCGGAG AAACCGACGT GGCGAAAATC 751 CTGTTGGGCG CGGGCTTGGG CATAACGGGC ATTCTGGCAG TCGTCCTCTC 801 CACCGTTACC ACAACGTTTC TCGATACCTA TTCCGCCGGC GCGAGTGCGA 851 ACAACATTTC CGCGCGTTTT GCGGAAATAC CCGTCGCTGT CGGCGTTACC 901 CTGATCGGCA CGGTGCTTGC CGTCATGCTG CCCGTTACCG AATATAAAAA 951 CTTCCTGCTG CTTATCGGCT CGGTATTTGC GCCGATGGCG GCGGTTTTGA 1001 TTGCCGACTT TTTCGTCTTA AAACGGCGTG AGGAGATTGA AGGCTTTGAC 1051 TTTGCCGGAC TGGTTCTGTG GCTGGCAGGC TTCATCCTCT ACCGCTTCCT 1101 GCTCTCGTCC GGTTGGGAAA GCAGCATCGG TCTGACCGCC CCCGTAATGT 1151 CTGCCGTTGC CATTGCCACC GTATCGGTAC GCCTTTTCTT TAAAAAAACC 1201 CAATCTTTAC AAAGGAACCC GTCATGA This coπesponds to the amino acid sequence <SEQ ID 808; ORF125ng-l>: 1 MSGNASSPSS SAAIGLVWFG AAVSIAEIST GTLLAPLGWQ RGLAALLLGH 51 AVGGALFFAA AYIGALTGRS SMESVRLSFG KCGSVLFSVA NMLQLAGWTA 101 VMIYVGATVS SALGKVLWDG ESFVWWALAN GALIVLWLVF GARRTGGLKT 151 VSMLLMLLAV LWLSVEVFAS SGTNAAPAVS DGMTFGTAVE LSAVMPLSWL 201 PLAADYTRQA RRPFAATLTA TLAYTLTGCW MYALGLAAAL FTGETDVAKI. 251 LLGAGLGITG ILAWLSTVT TTFLDTYSAG ASANNISARF AEIPVAVGVT 301 LIGTVLAVML PVTEYKNFLL LIGSVFAPMA AVLIADFFVL KRREEIEGFD 351 FAGLVLWLAG FILYRFLLSS GWESSIGLTA PVMSAVAIAT VSVRLFFKKT 401 QSLQRNPS* ORF125ng-l and ORF125-1 show 95.1% identity in 408 aa overlap: 10 20 30 40 50 60 orf125-1. pep MSGNASSPSSSSAIGLIWFGAAVSIAEISTGTLLAPLGWQRGLAALLLGHAVGGALFFAA I I I I II I I I II : I I I I : I I II II I II II II I I I II I I I I I I I I II I I I I I II I II I II I I orfl25ng-l MSGNASSPSSSAAIGLVWFGAAVSIAEISTGTLLAPLGWQRGLAALLLGHAVGGALFFAA 10 20 30 40 50 60 70 80 90 100 110 120 orf125-1. pep AYIGALTGRSSMESVRLSFGKRGSVLFSVANMLQLAGWTAVMIYAGATVSSALGKVLWDG I I I II I I I II I I II II I I II I I II I II II I I I II I I II I I I I I : I I I II II I I I I I I I I orfl25ng-l AYIGALTGRSSMESVRLSFGKCGSVLFSVANMLQLAGWTAVMIYVGATVSSALGKVLWDG 70 80 90 100 110 120 130 140 150 160 170 179 orf125-1. pep ESFVWWALANGALIVLWLVFGARKTGGLKTVSMLLMLLAVLWLSAEVFSTAGSTAAQ-VS I I I I I II II I II I II I I I I I I I I : I I I I I I II I II I I I I I I I I I : II I::: M: I I II orfl25ng-l ESFVWWALANGALIVLWLVFGARRTGGLKTVSMLLMLLAVLWLSVEVFASSGTNAAPAVS 130 140 150 160 170 180 180 190 200 210 220 230 239 orf125-1. pep DGMSFGTAVELSAVMPLSWLPLAADYTRHARRPFAATLTATLAYTLTGCWMYALGLAAAL II I : I I I I II I I II I I I I II I II I I I I I : I I I II I I I I II I I I I II II I II I II I II I II orfl25ng-l DGMTFGTAVELSAVMPLSWLPLAADYTRQARRPFAATLTATLAYTLTGCWMYALGLAAAL 190 200 210 220 230 240 240 250 260 270 280 290 299 orf125-1. pep FTGETDVAKILLGAGLGAAGILAWLSTVTTTFLDAYSAGASANNISARFAETPVAVGVT I II I II I I I I I I I I I I I :| I I I II I I I II II I IM I I I I II I I I I I II I I I I I I II I I orfl25ng-l FTGETDVAKILLGAGLGITGILAWLSTVTTTFLDTYSAGASANNISARFAEIPVAVGVT 250 260 270 280 290 300 300 310 320 330 340 350 359 orf125-1. pep LIGTVLAVMLPVTEYENFLLLIGSVFAPMAAVLIADFFVLKRREEIEGFDFAGLVLWLAG I II I II I II I I I I I I : I I I II I II II II II II II I I II I I I I I I I I I I II II I I II I I I I orfl25ng-l LIGTVLAVMLPVTEYKNFLLLIGSVFAPMAAVLIADFFVLKRREEIEGFDFAGLVLWLAG 310 320 330 340 350 360 360 370 380 390 400 orf125-1. pep FILYRFLLSSGWESSIGLTAPVMSAVAIATVSVRLFFKKTQSLQRNPSX I II I II I II I II I II I I II II I II I I I II I I I I I I I I I I I I I I II I I I I orfl25ng-l FILYRFLLSSGWESSIGLTAPVMSAVAIATVSVRLFFKKTQSLQRNPSX 370 380 390 400 Based on this analysis, including the presence of putative leader sequence and transmembrane domains in the gonococcal protein, it is predicted that the proteins from N. meningitidis .and N.gonorrhoeae, and their epitopes, could be useful antigens for vaccines or diagnostics, or for raising antibodies. Example 96 The following partial DNA sequence was identified in N. meningitidis <SEQ ID 809>: 1 ATGACCCGTA TCGCCATCCT CGGCGGCGGC CTCTCGGGAA GGCTGACCGC 51 GTTGCAGCTT GCAGAACAAG GTTATCAGAT TGCACTTTTC GATAAAAGCT 101 GCCGCCGGGG CGAACACGCC GCCGCCTATG TAGCCGCCGC CATGCTCGCG 151 CCTGCAGCGG A.ACGGTCGA AGCCACGCCC GAAGTGGTCA GGCTGGGCAG 201 GCAGAGCATC CCGCTTTGGC GCGGCATCCG ATGCCGTCTG AACACGCACA 251 CGATGATGCA GGAAAACGGC AGCCTGATTG TATGGCACGG GCAGGACAAG 301 CCATTATCCA GCGAGTTCGT CCGCCATCTC AAACGCGGCG GCGT.ACGGA 351 TGACGAAATC GTCCGTTGGC GCGCCGACGA CATCGCCGAA CGCGAACCGC 401 AACTCGGCGG ACGTTTTTAA GACGGCATCT ACCTGCCGAC CGAAGC.CAG 451 CTCGACGGGC GGCAATTATA GTCTGCACTT GCCGACGCTT TGGACGAACT 501 GAACGTCCCC TGCCATTGGG AACACGAATG CGTCCCCGAA GCCTGCAAG .. This coπesponds to the amino acid sequence <SEQ ID 810; ORF126>: 1 MTRIAILGGG LSGRLTALQL AEQGYQIALF DKSCRRGEHA AAYVAAAMLA 51 PAAXTVEATP E RLGRQSI PLWRGIRCRL NTHTMMQENG SLIVWHGQDK 101 PLSSEFVRHL KRGGXTDDEI VRWRADDIAE REPQLGGRFX DGIYLPTEXQ 151 LDGRQLXSAL ADALDELNVP CHWEHECVPE ACK... Further work revealed the complete nucleotide sequence <SEQ ID 811>: 1 ATGACCCGTA TCGCCATCCT CGGCGGCGGC CTCTCGGGAA GGCTGACCGC 51 GTTGCAGCTT GCAGAACAAG GTTATCAGAT TGCACTTTTC GATAAAGGCT 101 GCCGCCGGGG CGAACACGCC GCCGCCTATG TTGCCGCCGC CATGCTCGCG 151 CCTGCGGCGG AAGCGGTCGA AGCCACGCCC GAAGTGGTCA GGCTGGGCAG 201 GCAGAGCATC CCGCTTTGGC GCGGCATCCG ATGCCGTCTG AACACGCACA 251 CGATGATGCA GGAAAACGGC AGCCTGATTG TGTGGCACGG GCAGGACAAG 301 CCATTATCCA GCGAGTTCGT CCGCCATCTC AAACGCGGCG GCGTAGCGGA 351 TGACGAAATC GTCCGTTGGC GCGCCGACGA CATCGCCGAA CGCGAACCGC 401 AACTCGGCGG ACGTTTTTCA GACGGCATCT ACCTGCCGAC CGAAGGCCAG 451 CTCGACGGGC GGCAAATATT GTCTGCACTT GCCGACGCTT TGGACGAACT 501 GAACGTCCCC TGCCATTGGG AACACGAATG CGTCCCCGAA GGCCTGCAAG 551 CCCAATACGA CTGGCTGATC GACTGCCGCG GCTACGGCGC AAAAACCGCG 601 TGGAACCAAT CCCCCGAGCA CACCAGCACC CTGCGCGGCA TACGCGGCGA 651 AGTGGCGCGG GTTTACACAC CCGAAATCAC GCTCAACCGC CCCGTGCGTC 701 TGCTCCATCC GCGTTATCCG CTCTACATCG CCCCGAAAGA AAACCACGTC 751 TTCGTCATCG GCGCGACCCA AATCGAAAGC GAAAGCCAAG CCCCCGCCAG 801 CGTGCGTTCA GGGTTGGAAC TCTTGTCCGC ACTCTATGCC ATCCACCCCG 851 CCTTCGGCGA AGCCGACATC CTCGAAATCG CCACCGGCCT GCGCCCCACG 901 CTCAACCACC ACAACCCCGA AATCCGTTAC AACCGCGCCC GACGCCTGAT 951 TGAAATCAAC GGCCTTTTCC GCCACGGTTT CATGATCTCC CCCGCCGTAA 1001 CCGCCGCCGC CGCCAGATTG GCAGTGGCAC TGTTTGACGG AAAAGACGCG 1051 CCCGAACGCG ATAAAGAAAG CGGTTTGGCG TATATCCGAA GACAAGATTA 1101 A This coπesponds to the amino acid sequence <SEQ ID 812; ORF126-l>: 1 MTRIAILGGG LSGRLTALQL AEQGYQIALF DKGCRRGEHA AAYVAAAMLA 51 PAAEAVEATP EWRLGRQSI PLWRGIRCRL NTHTMMQENG SLIVWHGQDK 101 PLSSEFVRHL KRGGVADDEI VRWRADDIAE REPQLGGRFS DGIYLPTEGQ 151 LDGRQILSAL ADALDELNVP CHWEHECVPE GLQAQYDWLI DCRGYGAKTA 201 WNQSPEHTST LRGIRGEVAR VYTPEITLNR PVRLLHPRYP LYIAPKENHV 251 FVIGATQIES ESQAPASVRS GLELLSALYA IHPAFGEADI LEIATGLRPT 301 LNHHNPEIRY NRARRLIEIN GLFRHGFMIS PAVTAAAARL AVALFDGKDA 351 PERDKESGLA YIRRQD* Computer analysis of this amino acid sequence gave the following results: Homology with a predicted ORF from N.meninsitidis (strain Al ORF 126 shows 90.0% identity over a 180aa overlap with an ORF (ORF 126a) from strain A of N meningitidis: 10 20 30 40 50 60 orf126. pep MTRIAILGGGLSGRLTALQLAEQGYQIALFDKSCRRGEHAAAYVAAAMLAPAAXTVEATP I I II II II II I I II I I I I I I I I II II I I I I I I : I I I I I I I I I I I I I I I I II I I MMM orf126a MTRIAILGGGLSGRLTALQLAEQGYQIALFDKGCRRGEHAAAYVAAAMLAPAAEAVEATP 10 20 30 40 50 60 70 80 90 100 110 120 orf126. pep EWRLGRQSIPLWRGIRCRLΝTHTMMQEΝGSLIVWHGQDKPLSSEFVRHLKRGGXTDDEI I II I II I I I I I II I I I I: I: I Ml I I I II I II I I II I M I M II II II II I Ml I orf126a EWRLGRQXIPLWRGIRCHLKTPAMMXEΝGSLIVWHGQDKPLSΝEFVRHLKRGGVADDXI 70 80 90 100 110 120 130 140 150 160 170 180 orf126. pep VRWRADDIAEREPQLGGRFXDGIYLPTEXQLDGRQLXSALADALDELΝVPCHWEHECVPE II I II I I I II II II I I I I I I I II II I I IIIIIM I I II I II I II I I I I I I I I I I M I orf126a VRWRADDIAEREPQLGGRFSDGIYLPTEGQLDGRQILSALADALDELΝVPCHWEHECAPE 130 140 150 160 170 180 The complete length ORF126a nucleotide sequence <SEQ ID 813> is: 1 ATGACCCGTA TCGCCATCCT CGGCGGCGGC CTCTCΝGGAA GGCTGACCGC 51 ACTGCAGCTT GCAGAACAAG GTTATCAGAT TGCACTTTTC GATAAAGGCT 101 GCCGCCGGGG CGAACACGCC GCCGCCTATG TTGCCGCCGC CATGCTCGCG 151 CCTGCGGCGG AAGCGGTCGA AGCCACGCCT GAAGTGGTCA GGCTGGGCAG 201 GCAGAΝCATC CCGCTTTGGC GCGGCATCCG ATGCCATCTG AAAACGCCTG 251 CCATGATGCA ΝGAAAACGGC AGCCTGATTG TGTGGCACGG GCAGGACAAA 301 CCTTTATCCA ACGAGTTCGT CCGCCATCTC AAACGCGGCG GCGTAGCGGA 351 TGACΝAAATC GTCCGTTGGC GCGCCGACGA CATCGCCGAA CGCGAACCGC 401 AACTCGGCGG ACGTTTTTCA GACGGCATCT ACCTGCCGAC CGAAGGCCAG 451 CTCGACGGGC GGCAAATATT GTCTGCACTT GCCGACGCTT TGGACGAACT 501 GAACGTCCCC TGCCATTGGG AACACGAATG TGCCCCCGAA GACTTGCAAG 551 CCCAATACGA CTGGCTGATC GACTGCCGCG GCTACGGCGC AAAAACCGCG 601 TGGAACCAAT CCCCCGAΝΝA ΝACCAGCACC CTGCGCGGCA TACGCGGCGA 651 AGTGGCGCGG GTTTACACAC CCGAAATCAC GCTCAACCGC CCCGTGCGCC 701 TGCTACACCC GCGCTATCCG CTΝTACATCG CCCCGAAAGA AAACCΝCGTC 751 TTCGTCATCG GCGCGACCCA AATCGAAAGC GAAAGCCAAG CACCTGCCAG 801 CGTGCGTTCC GGGCTGGAAC TCTTATCCGC ACTCTATGCC GTCCACCCCG 851 CCTTCGGCGA AGCCGACATC CTCGAAATCG CCACCGGCCT GCGCCCCACG 901 CTCAATCACC ACAACCCCGA AATCCGTTAC AACCGCGCCC GACGCCTGAT 951 TGAAATCAAC GGCCTTTTCC GCCACGGTTT CATGATCTCC CCCGCCGTAA 1001 CCGCCGCCGC CGTCAGATTG GCAGTGGCAC TGTTTGACGG AAAAGAΝGCG 1051 CCCGAACGCG ATGAAGAAAG CGGTTTGGCG TATATCCGAA GACAAGATTA 1101 A This encodes a protein having amino acid sequence <SEQ ED 814>: 1 MTRIAILGGG LSGRLTALQL AEQGYQIALF DKGCRRGEHA AAYVAAAMLA 51 PAAEAVEATP EWRLGRQXI PLWRGIRCHL KTPAMMXEΝG SLIVWHGQDK 101 PLSΝEFVRHL KRGGVADDXI VRWRADDIAE REPQLGGRFS DGIYLPTEGQ 151 LDGRQILSAL ADALDELNVP CHWEHECAPE DLQAQYDWLI DCRGYGAKTA 201 WNQSPXXTST LRGIRGEVAR VYTPEITLNR PVRLLHPRYP LYIAPKENXV 251 FVIGATQIES ESQAPASVRS GLELLSALYA VHPAFGEADI LEIATGLRPT 301 LNHHNPEIRY NRARRLIEIN GLFRHGFMIS PAVTAAAVRL AVALFDGKXA 351 PERDEESGLA YIRRQD* ORF 126a and ORF 126-1 show 95.4% identity in 366 aa overlap: 10 20 30 40 50 60 orf126a . pep MTRIAILGGGLSGRLTALQLAEQGYQIALFDKGCRRGEHAAAYVAAAMLAPAAEAVEATP II I II II I II II II II I II II I II I II I II II I I II I II I II I I I I I I II I I I I I I I I II orf126-1 MTRIAILGGGLSGRLTALQLAEQGYQIALFDKGCRRGEHAAAYVAAAMLAPAAEAVEATP 10 20 30 40 50 60 70 80 90 100 110 120 orf 126a . pep EWRLGRQXIPLWRGIRCHLKTPAMMXENGSLIVWHGQDKPLSNEFVRHLKRGGVADDXI I I I I I I I I I I I I I I I I I : I : I M l I I I I I I I I I I I I I I I I : I I I I I I I I I I I I I I I orf 126-1 EWRLGRQSIPLWRGIRCRLNTHTMMQENGSLIVWHGQDKPLSSEFVRHLKRGGVADDEI 70 80 90 100 110 120 130 140 150 160 170 180 or f 126a . pep VRWRADDIAEREPQLGGRFSDGIYLPTEGQLDGRQILSALADALDELNVPCHWEHECAPE I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I M I I orf126-1 VRWRADDIAEREPQLGGRFSDGIYLPTEGQLDGRQILSALADALDELNVPCHWEHECVPE 130 140 150 160 170 180 190 200 210 220 230 240 orf126a. pep DLQAQYDWLIDCRGYGAKTAWNQSPXXTSTLRGIRGEVARVYTPEITLNRPVRLLHPRYP II I I I II I II II I II II I II I II I II II II I I II I II I I I II I I I II I I I I I I I I II orf126-1 GLQAQYDWLIDCRGYGAKTAWNQSPEHTSTLRGIRGEVARVYTPEITLNRPVRLLHPRYP 190 200 210 220 230 240 250 260 270 280 290 300 orf126a . pep LYIAPKENXVFVIGATQIESESQAPASVRSGLELLSALYAVHPAFGEADILEIATGLRPT I I II I I I I I I I II II I II I I I I I I I I I I I I I II I I I I I I: I II II I I I I I II II II I I I orf126-1 LYIAPKENHVFVIGATQIESESQAPASVRSGLELLSALYAIHPAFGEADILEIATGLRPT 250 260 270 280 290 300 310 320 330 340 350 360 orf126a. pep LNHHNPEIRYNRARRLIEINGLFRHGFMISPAVTAAAVRLAVALFDGKXAPERDEESGLA I I II II I II I I I II I II I I I I I I I II II II II I II I I : I I I II I I I I I II I I I : I II II orf126-1 LNHHNPEIRYNRARRLIEINGLFRHGFMISPAVTAAAARLAVALFDGKDAPERDKESGLA 310 320 330 340 350 360 orf126a. pep YIRRQDX I II I I I I orf126-1 YIRRQDX Homology with a predicted ORF from Nεonorrhoeae ORF126 shows 90% identity over a 180 aa overlap with a predicted ORF (ORF126ng) from N.gonorrhoeae: orf126. pep MTRIAILGGGLSGRLTALQLAEQGYQIALFDKSCRRGEHAAAYVAAAMLAPAAXTVEATP 60 I I II I : I I II I II I II II I II I I II I I MM: I : I I I II I II I I I II I I I I MMM orfl26ng MTRIAVLGGGLSGRLTALQLAEQGYQIELFDKGTRQGEHAAAYVAAAMLAPAAEAVEATP 60 orf126. pep EWRLGRQSIPLWRGIRCRLNTHTMMQENGSLIVWHGQDKPLSSEFVRHLKRGGXTDDEI 120 II : I I I II I I II I II II I I II I I II I I II II I II I I II I I I I I I II I II I I I I Mill orfl26ng EVIRLGRQSIPLWRGIRCRLNTLTMMQENGSLIVWHGQDKPLSSEFVRHLKRGGVADDEI 120 orf126. pep VRWRADDIAEREPQLGGRFXDGIYLPTEXQLDGRQLXSALADALDELNVPCHWEHECVPE 180 I I II I I : I II I I I I II I II II I II I I I IIIIIM I I I II I II I I I I I II I I I I I : I : orfl26ng VRWRADEIAEREPQLGGRFSDGIYLPTEGQLDGRQILSALADALDELNVPCHWEHECAPQ 180 An ORF126ng nucleotide sequence <SEQ LD 815> was predicted to encode a protein having amino acid sequence <SEQ ED 816>: 1 MTRIAVLGGG LSGRLTALQL AEQGYQIELF DKGTRQGEHA AAYVAAAMLA 51 PAAEAVEATP EVIRLGRQSI PLWRGIRCRL NTLTMMQENG SLIVWHGQDK 101 PLSSEFVRHL KRGGVADDEI VRWRADEIAE REPQLGGRFS DGIYLPTEGQ 151 LDGRQILSAL ADALDELNVP CHWEHECAPQ DLQAQYDWVI DCRGYGAKTA 201 WNQSPEHTST LRGIRGEVRG FTRPKSRSTA PCACCTRAIR STSPRKKTTS 251 SSSARPKSKA KAKPPPAYVP GWNSYPRSMP STPPSAKPTS SKWRPGLRPT 301 LNHHNPEIRY SRERRLIEIN GLFRHGFMIS PAVTAAAVRL AVALFDGKDA 351 PERDEESGLA YIGRQD* Further work revealed the following gonococcal DNA sequence <SEQ ID 817>: 1 ATGACCCGTA TCGCCGTCCT CGGAGGCGGC CTTTCCGGAA GGCTGACCGC 51 ATTGCAGCTT GCAGAACAAG GTTATCAGAT TGAACTTTTC GACAAGGGCA 101 CCCGCCAAGG CGAACACGCC GCCGCCTATG TTGCCGCCGC GATGCTCGCG 151 CCTGCGGCGG AAGCGGTCGA GGCAACGCCC GAAGTCATCA GGCTGGGCAG 201 GCAGAGCATT CCGCTTTGGC GCGGCATCCG ATGCCGTCTG AACACGCTCA 251 CGATGATGCA GGAAAACGGC AGCCTGATTG TGTGGCACGG GCAGGACAAG 301 CCATTATCCA GCGAGTTCGT CCGCCATCTC AAACGCGGCG GCGTAGCGGA 351 TGACGAAATC GTCCGTTGGC GCGCCGATGA AATCGCCGAA CGCGAACCGC 401 AACTCGGCGG ACGTTTTTCA GACGGCATCT ACCTGCCGAC CGAAGGCCAG 451 CTCGACGGGC GGCAAATATT GTCTGCACTT GCCGACGCTT TGGACGAACT 501 GAACGTCCCT TGCCATTGGG AACACGAATG CGCCCCCCAA GACCTGCAAG 551 CCCAATACGA CTGGGTAATC GACTGCCGGG GCTACGGCGC GAAAACCGCG 601 TGGAACCAAT CCCCCGAGCA CACCAGCACC TTGCGCGGCA TACGCGGCGA 651 AGTGGCGCGG GTTTACACGC CCGAAATCAC GCTCAACCGC CCCGTGCGCC 701 TGCTGCACCC GCGCTATCCG CTCTACATCG CCCCGAAAGA AAACCACGTC 751 TTCGTCATCG GCGCGACCCA AATCGAAAGC GAAAGCCAAG CCCCCGCCAG 801 CGTACGTTCC GGGCTGGAAC TCTTATCCGC GCTCTATGCC GTCCACCCCG 851 CCTTCGGCGA AGCCGACATC CTCGAAATCG CCGCCGGCCT GCGCCCCACG 901 CTCAACCACC ACAACCCCGA AATCCGCTAC AGCCGCGAAC GCCGCCTCAT 951 CGAAATCAAC GGCCTTTTCC GGCACGGCTT TATGATTTCC CCCGCCGTAA 1001 CCGCCGCCGC CGTCAGATTG GCAGTGGCAC TGTTTGACGG AAAAGACGCG 1051 CCCGAACGTG ATGAAGAAAG CGGTTTGGCG TATATCGGAA GACAAGATTA 1101 A This coπesponds to the amino acid sequence <SEQ ED 818; ORF126ng-l>: 1 MTRIAVLGGG LSGRLTALQL AEQGYQIELF DKGTRQGEHA AAYVAAAMLA 51 PAAEAVEATP EVIRLGRQSI PLWRGIRCRL NTLTMMQENG SLIVWHGQDK 101 PLSSEFVRHL KRGGVADDEI VRWRADEIAE REPQLGGRFS DGIYLPTEGQ 151 LDGRQILSAL ADALDELNVP CHWEHECAPQ DLQAQYDWVI DCRGYGAKTA 201 WNQSPEHTST LRGIRGEVAR VYTPEITLNR PVRLLHPRYP LYIAPKENHV 251 FVIGATQIES ESQAPASVRS GLELLSALYA VHPAFGEADI LEIAAGLRPT 301 LNHHNPEIRY SRERRLIEIN GLFRHGFMIS PAVTAAAVRL AVALFDGKDA 351 PERDEESGLA YIGRQD* ORF126ng-l and ORF126-1 show 95.1% identity in 366 aa overlap: 10 20 30 40 50 60 orf126-1. pep MTRIAILGGGLSGRLTALQLAEQGYQIALFDKGCRRGEHAAAYVAAAMLAPAAEAVEATP I I II M II I II I II II I II I I I I I I I I I I II I I: I II I I I I I I I I II I I II I I II II I orfl26ng-l MTRIAVLGGGLSGRLTALQLAEQGYQIELFDKGTRQGEHAAAYVAAAMLAPAAEAVEATP 10 20 30 40 50 60 70 80 90 100 110 120 orf 126-1 . pep EWRLGRQSIPLWRGIRCRLNTHTMMQENGSLIVWHGQDKPLSSEFVRHLKRGGVADDEI I I : I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I orf l26ng-l EVIRLGRQSIPLWRGIRCRLNTLTMMQENGSLIVWHGQDKPLSSEFVRHLKRGGVADDEI 70 80 90 100 110 120 130 140 150 160 170 180 orf126-1. pep VRWRADDIAEREPQLGGRFSDGIYLPTEGQLDGRQILSALADALDELNVPCHWEHECVPE I II I I I: I I II I I I I II I II I I I I I II I I II I II I I I I I I II II II I I II II I II II : I: orfl26ng-l VRWRADEIAEREPQLGGRFSDGIYLPTEGQLDGRQILSALADALDELNVPCHWEHECAPQ 130 140 150 160 170 180 190 200 210 220 230 240 orf126-1. pep GLQAQYDWLIDCRGYGAKTAWNQSPEHTSTLRGIRGEVARVYTPEITLNRPVRLLHPRYP I I II I I I: I I II I II II I I I I I I I I I I II I I II I I I I I I I I II I I I I II II I II II I I I orfl26ng-l DLQAQYDWVIDCRGYGAKTAWNQSPEHTSTLRGIRGEVARVYTPEITLNRPVRLLHPRYP 190 200 210 220 230 240 250 260 270 280 290 300 orf126-1. pep LYIAPKENHVFVIGATQIESESQAPASVRSGLELLSALYAIHPAFGEADILEIATGLRPT I II II II I I I I II II I I I I II I I I II I I I II I II II I II I: I I I I I I I I II I I M I I II I orfl26ng-l LYIAPKENHVFVIGATQIESESQAPASVRSGLELLSALYAVHPAFGEADILEIAAGLRPT 250 260 270 280 290 300 310 320 330 340 350 360 orf126-1. pep LNHHNPEIRYNRARRLIEINGLFRHGFMISPAVTAAAARLAVALFDGKDAPERDKESGLA I II II I I I I I : I II I I I I I II II I I II II II I I I I I : I I I I I II I I I I I I I I I : I I I I I orfl26ng-l LNHHNPEIRYSRERRLIEINGLFRHGFMISPAVTAAAVRLAVALFDGKDAPERDEESGLA 310 320 330 340 350 360 orf126-1. pep YIRRQDX 11 1111 orfl26ng-l YIGRQDX Furthermore, ORF126ng-l shows homology to a putative Rhizobium oxidase flavoprotein: gi 12627327 (AF004408) putative amino acid oxidase flavoprotein [Rhizobium etli] Length = 327 Score = 169 bits (423), Expect = 3e-41 Identities = 112/329 (34%), Positives = 163/329 (49%), Gaps = 25/329 (7%) Query: 3 RIAVLGGGLSGRLTALQLAEQGYQIELFDKGTRQGEHXXXXXXXXXXXXXXXXXXXXXXX 62 RI V G G++G A QL G+++ L ++ G Sbjct: 2 RILVNGAGVAGLTVAWQLYRHGFRVTLAERAGTVGA-GASGFAGGMLAPWCERESAEEPV 60 Query: 63 IRLGRQSIPLWRGIRCRLNTLTMMQENGSLIVWHGQDKPLSSEFVRHLKRGGVADDEIVR 122 + LGR + W + G+L+V G+D F R G DE+ Sbjct: 61 LTLGRLAADWWEAA LPGHVHRRGTLWAGGRDTGELDRFSRRTS-GWEWLDEVA- 113 Query: 123 WRADEIAEREPQLGGRFSDGIYLPTEGQLDGRQILSALADALDELNVPCHWEHECAPQDL 182 IA EP L GRF ++ E LD RQ L+ALA L++ + + Sbjct: 114 IAALEPDLAGRFRRALFFRQEAHLDPRQALAALAAGLEDARMRLTLG WGES 165 Query: 183 QAQYDWVIDCRGYGAKTAWNQSPEHTSTLRGIRGEVARVYTPEITLNRPVRLLHPRYPLY 242 +D V+DC G LRG+RGE+ V T E++L+RPVRLLHPR+P+Y Sbjct: 166 DVDHDRWDCTGAA QIGRLPGLRGVRGEMLCVETTEVSLSRPVRLLHPRHPIY 218 Query: 243 IAPKENHVFVIGATQIESESQAPASVRSGLELLSALYAVHPAFGEADILEIAAGLRPTLN 302 I P++ + F++GAT IES+ P + RS +ELL+A YA+HPAFGEA + E AG+RP Sbjct: 219 IVPRDKNRFMVGATMIESDDGGPITARSLMELLNAAYAMHPAFGEARVTETGAGVRPAYP 278 Query: 303 HHNPEIRYSRERRLIEINGLFRHGFMISP 331 + P R ++E R + +NGL+RHGF+++P Sbjct: 279 DNLP--RVTQEGRTLHVNGLYRHGFLLAP 305 This analysis suggests that the proteins from Nmeningitidis and N.gonorrhoeae, and their epitopes, could be useful .antigens for vaccines or diagnostics, or for raising antibodies. Example 97 The following DNA sequence, believed to be complete, was identified in Nmeningitidis <SEQ ID 819>: 1 ATGACTGATA ATCGGGGGTT TACGCTGGTT GAATTAATAT CAGTGGTCTT 51 GATATTGTCT GTACTTGCTT TAATTGTTTA TCCGAGCTAT CGCAATTATG 101 TTGAGAAAGC AAAGATAAAT GCAGTGCGGG CAGCCTTGTT AGAAAATGCA 151 CATTTTATGG AAAAGTTTTA TCTGCAGAAT GGGAGGTTTA AACAAACATC 201 TACCAAGTGG CCAAGTTTGC CGATTAAAGA GGCAGAAGGC TTTTGTATCC 251 GTTTGAATGG AATCGtCGCG CGGG..GCTT TAGACAGTAA ATTCATGTTG 301 AAGGCGGTAG CCATAGATAA AGATAAAAAT CCTTTTATTA TTAAGATGAA 351 TGAAAATCTA GTAACCTTTA aTTTGCAAGA AGTCCGCCAG TTCGTGTAGT 401 GACGGGCTGG ATTATTTTAA AGGAAATGAT AAGGACTGCA AGTTACTTAA 451 GTAG This coπesponds to the amino acid sequence <SEQ ID 820; ORF127>: 1 MTDNRGFTLV ELISWLILS VLALIVYPSY RNYVEKAKIN AVRAALLENA 51 HFMEKFYLQN GRFKQTSTKW PSLPIKEAEG FCIRLNGIVA RXALDSKFML 101 KAVAIDKDKN PFIIKMNENL VTFICKKSAS SCSDGLDYFK GNDKDCKLLK 151 * Further work revealed the following DNA sequence <SEQ ID 821>: 1 ATGACTGATA ATCGGGGGTT TACGCTGGTT GAATTAATAT CAGTGGTCTT 51 GATATTGTCT GTACTTGCTT TAATTGTTTA TCCGAGCTAT CGCAATTATG 101 TTGAGAAAGC AAAGATAAAT GCAGTGCGGG CAGCCTTGTT AGAAAATGCA 151 CATTTTATGG AAAAGTTTTA TCTGCAGAAT GGGAGGTTTA AACAAACATC 201 TACCAAGTGG CCAAGTTTGC CGATTAAAGA GGCAGAAGGC TTTTGTATCC 251 GTTTGAATGG AATCGCGCGC GGGGCTTTAG ACAGTAAATT CATGTTGAAG 301 GCGGTAGCCA TAGATAAAGA TAAAAATCCT TTTATTATTA AGATGAATGA 351 AAATCTAGTA ACCTTTATTT GCAAGAAGTC CGCCAGTTCG TGTAGTGACG 401 GGCTGGATTA TTTTAAAGGA AATGATAAGG ACTGCAAGTT ACTTAAGTAG This coπesponds to the amino acid sequence <SEQ ID 822; ORF127-l>: 1 MTDNRGFTLV ELISWLILS VLALIVYPSY RNYVEKAKIN AVRAALLENA 51 HFMEKFYLQN GRFKQTSTKW PSLPIKEAEG FCIRLNGIAR GALDSKFMLK 101 AVAIDKDKNP FIIKMNENLV TFICKKSASS CSDGLDYFKG NDKDCKLLK* Computer analysis of this amino acid sequence gave the following results: Homology with a predicted ORF from N. meningitidis (strain A) ORF127 shows 98.0% identity over a 150aa overlap with an ORF (ORF127a) from sfrain A of N. meningitidis: 10 20 30 40 50 60 orf127.pep MTDNRGFTLVELISWLILSVLALIVYPSYRNYVEKAKINAVRAALLENAHFMEKFYLQN I II I I I I I II I I II I II I I I I I II I I II I I I I II I I I I I I : I I I II II I I I II I II II I I orf127a MTDNRGFTLVELISWLILSVLALIVYPSYRNYVEKAKINTVRAALLENAHFMEKFYLQN 10 20 30 40 50 60 70 80 90 100 110 120 orf127.pep GRFKQTSTKWPSLPIKEAEGFCIRLNGIVARXALDSKFMLKAVAIDKDKNPFIIKMNENL I II I I II II I I II I I I I I I I I I II I I I I II II I I I I I I I I I I II II I I I II I II II I I orf127a GRFKQTSTKWPSLPIKEAEGFCIRLNGI-ARGALDSKFMLKAVAIDKDKNPFIIKMNENL 70 80 90 100 110 130 140 150 orf127.pep VTFICKKSASSCSDGLDYFKGNDKDCKLLKX II I II I I I I II I I I II II I II II I II I I I II orf127a VTFICKKSASSCSDGLDYFKGNDKDCKLLKX 120 130 140 150 The complete length ORF 127a nucleotide sequence <SEQ ID 823> is: 1 ATGACTGATA ATCGGGGGTT TACGCTGGTT GAATTAATAT CAGTGGTCTT 51 GATATTGTCT GTACTTGCTT TAATTGTTTA TCCGAGCTAT CGCAATTATG 101 TTGAGAAAGC AAAGATAAAT ACAGTGCGGG CAGCCTTGTT AGAAAATGCA 151 CATTTTATGG AAAAGTTTTA TCTGCAGAAT GGGAGATTTA AACAAACATC 201 TACCAAATGG CCAAGTTTGC CGATTAAAGA GGCAGAAGGC TTTTGTATCC 251 GTTTGAATGG AATCGCGCGC GGGGCCTTAG ACAGTAAATT CATGTTGAAG 301 GCGGTAGCCA TAGATAAAGA TAAAAATCCT TTTATTATTA AGATGAATGA 351 AAATCTAGTA ACCTTTATTT GCAAGAAGTC CGCCAGTTCG TGTAGTGACG 401 GGCTGGATTA TTTTAAAGGA AATGATAAGG ACTGCAAGTT ACTTAAGTAG This encodes a protein having amino acid sequence <SEQ ID 824>: 1 MTDNRGFTLV ELISWLILS VLALIVYPSY RNYVEKAKIN TVRAALLENA 51 HFMEKFYLQN GRFKQTSTKW PSLPIKEAEG FCIRLNGIAR GALDSKFMLK 101 AVAIDKDKNP FIIKMNENLV TFICKKSASS CSDGLDYFKG NDKDCKLLK* ORF127a and ORF127-1 show 99.3% identity in 149 aa overlap: 10 20 30 40 50 60 orf127a. pep MTDNRGFTLVELISWLILSVLALIVYPSYRNYVEKAKINTVRAALLENAHFMEKFYLQN II I I II II II II I II II II I I I I I I II II II I II I I I II M I II I I II I I I I II II I I I I orfl27-l MTDNRGFTLVELISWLILSVLALIVYPSYRNYVEKAKINAVRAALLENAHFMEKFYLQN 10 20 30 40 50 60 70 80 90 100 110 120 orf127a. pep GRFKQTSTKWPSLPIKEAEGFCIRLNGIARGALDSKFMLKAVAIDKDKNPFIIKMNENLV I I I I I I II I I I I I II II I I I I I I I I I II I I II I II I I I I M I I I I I I I I I II I I I I I II I orf127-1 GRFKQTSTKWPSLPIKEAEGFCIRLNGIARGALDSKFMLKAVAIDKDKNPFIIKMNENLV 70 80 90 100 110 120 130 140 150 orfl27a.pep TFICKKSASSCSDGLDYFKGNDKDCKLLKX II I I I II II I I I I I I I I I I I I I I I I I I II I orfl27-l TFICKKSASSCSDGLDYFKGNDKDCKLLKX 130 140 150 Homology with a predicted ORF from N.εonorrhoeae ORF127 shows 97.3% identity over a 150 aa overlap with a predicted ORF (ORF127ng) from N.gonorrhoeae: orf127.pep MTDNRGFTLVELISWLILSVLALIVYPSYRNYVEKAKINAVRAALLENAHFMEKFYLQN 60 I II I I I I I II I I II II I I II I II I I II II II I I I I I I I II I I I II: I I I I I I I II II II I orfl27ng MTDNRGFTLVELISWLILSVLALIVYPSYRNYVEKAKINAVRAAFLENAHFMEKFYLQN 60 orf127.pep GRFKQTSTKWPSLPIKEAEGFCIRLNGIVARXALDSKFMLKAVAIDKDKNPFIIKMNENL 120 I I II I II II I I I I I I II II I I I I I I I II II I I II I II I I II I I I II I I II II I I I I I I orfl27ng GRFKQTSTKWPSLPIKEAEGFCIRLNGI-ARGALDSKFMLKAVAIDKDKNPFIIKMNENL 119 orf127. pep VTFICKKSASSCSDGLDYFKGNDKDCKLLK 150 II I I I I I I II I I II I I I I I I I II I I I I I I orfl27ng VTFICKKSASSCSDRLDYFKGNDKDCKLLK 149 The complete length ORF127ng nucleotide sequence <SEQ ID 825> is: 1 ATGACTGATA ATCGGGGGTT TACACTGGTT GAATTAATAT CAGTGGTCTT 51 GATATTGTCT GTACTTGCTT TAATTGTTTA TCCGAGCTAT CGCAATTATG 101 TTGAGAAAGC AAAGATAAAT GCAGTGCGGG CAGCCTTGTT AGAAAATGCA 151 CATTTTATGG AAAAGTTTTA TCTGCAGAAT GGGAGATTTA AACAAACATC 201 TACCAAATGG CCAAGTTTGC CGATTAAAGA GGCAGAAGGC TTTTGTATCC 251 GTTTGAATGG AATCGCGCGC GGGGCTTTAG ACAGTAAATT CATGTTGAAG 301 GCGGTAGCCA TAGATAAAGA TAAAAATCCT TTTATTATTA AGATGAATGA 351 AAATCTAGTA ACCTTTATTT GCAAGAAGTC CGCCAGTTCG TGTAGTGACG 401 GGCTGGATTA TTTTAAAGGA AATGATAAGG ACTGCAAGTT ACTTAAGTAG This encodes a protein having amino acid sequence <SEQ LD 826>: 1 MTDNRGFTLV ELISWLILS VLALIVYPSY RNYVEKAKIN AVRAAFLENA 51 HFMEKFYLQN GRFKQTSTKW PSLPIKEAEG FCIRLNGIAR GALDSKFMLK 101 AVAIDKDKNP FIIKMNENLV TFICKKSASS CSDRLDYFKG NDKDCKLLK* ORF127ng and ORF127-1 show 100.0% identity in 149 aa overlap: 10 20 30 40 50 60 orf127-1.pep MTDNRGFTLVELISWLILSVLALIVYPSYRNYVEKAKINAVRAALLENAHFMEKFYLQN I II I I I I II I I I I I II II I I I I I II I II II I I II I I I II I I I I I I I I I I I I II I II I I I I orfl27ng-l MTDNRGFTLVELISWLILSVLALIVYPSYRNYVEKAKINAVRAALLENAHFMEKFYLQN 10 20 30 40 50 60 70 80 90 100 110 120 orf127-1. pep GRFKQTSTKWPSLPIKEAEGFCIRLNGIARGALDSKFMLKAVAIDKDKNPFIIKMNENLV III II II I II II II I I I II I II I I I I I I I I II I I II I I II II I I I I I I II II I I I I I II I orfl27ng-l GRFKQTSTKWPSLPIKEAEGFCIRLNGIARGALDSKFMLKAVAIDKDKNPFIIKMNENLV 70 80 90 100 110 120 130 140 150 orf127-1. pep TFICKKSASSCSDGLDYFKGNDKDCKLLKX I II I I I I I II I I I I I I II I II I I I I I I II I orfl27ng-l TFICKKSASSCSDGLDYFKGNDKDCKLLKX 130 140 150 This analysis, including the fact that the predicted transmembrane domain is shared by the meningococcal and gonococcal proteins, suggests that the proteins from N. meningitidis and N.gonorrhoeae, and their epitopes, could be useful antigens for vaccines or diagnostics, or for raising antibodies. Example 98 The following partial DNA sequence was identified in N. meningitidis <SEQ ID 827> 1 ..GTGTCGCTGG CTTCGGTGAT TGCCTCTCAA ATCTTCCTTT ACGAAGATTT 51 CAACCAAATG CGGAAAACC£ GTGGAGCTAT CTGCGGTTTT CTTGTCCAAT 101 ATTTATCTGG GGTTTCAGCA GGGGTATTTC GATTTGAGTG CCGACGAGAA 151 CCCCGTACTG CATATCTGGT CTTTGGCAGT AGAGGAACAG TATTACCTCC 201 TGTATCCCCT TTTGCTGATA TTTTGCTGCA AAAAAACCAA ATCGCTACGG 251 GTGCTGCGTA ACATCAGCAT CATCCTGTTT TTGATTTTGA CTGCCTCATC 301 GTTTTTGCCA AGCGGGTTTT ATACCGACAT CCTCAACCAA CCCAATACTT 351 ATTACCTTTC GACACTGAGG TTTCCCGAGC TGTTGGCAGG TTCGCTGCTG 401 GCGGTTTACG GGCAAACGCA AAACGGCAGA CGGCAAACAG CAAATGGAAA 451 ACGGCAGTTG CTTTCATCAC TCTGCTTCGG CGCATTGCTT GCCTGCCTGT 501 TCGTGATTGA CAAACACAAT CCGTTTATCC CGGGAATGAC CCTGCTCCTT 551 CCCTGCCTGC TGACGGCACT GCTTATCCGG AGTATGCAAT ACGGGACACT 601 TCCGACCCGC ATCCTGTCGG CAAGCCCCAT CGTATTTGTC GGCAAAATCT 651 CTTATTCCCT ATACCTGTAC CATTGGATTT TTATTGCTTT CGCTCCGCTC 701 ATTAGAGGCG GGAAACAGCT CGGACTGCCT GCCG.. This coπesponds to the amino acid sequence <SEQ ID 828; ORF128>: 1 .. VSLASVIASQ IFLYEDFNQM RKTVELSAVF LSNIYLGFQQ GYFDLSADEN 51 PVLHIWSLAV EEQYYLLYPL LLIFCCKKTK SLRVLRNISI ILFLILTASS 101 FLPSGFYTDI LNQPNTYYLS TLRFPELLAG SLLAVYGQTQ NGRRQTANGK 151 RQLLSSLCFG ALLACLFVID KHNPFIPGMT LLLPCLLTAL LIRSMQYGTL 201 PTRILSASPI VFVGKISYSL YLYHWIFIAF APLIRGGKQL GLPA. . Further work revealed the complete nucleotide sequence <SEQ ID 829>: 1 ATGCAAGCTG TCCGATACAG ACCGGAAATT GACGGATTGC GGGCCGTCGC 51 CGTGCTATCC GTCATGATTT TCCACCTGAA TAACCGCTGG CTGCCCGGAG 101 GATTCCTGGG GGTGGACATT TTCTTTGTCA TCTCAGGATT CCTCATTACC 151 GGCATCATTC TTTCTGAAAT ACAGAACGGT TCTTTTTCTT TCCGGGATTT 201 TTATACCCGC AGGATTAAGC GGATTTATCC TGCCTTTATT GCGGCCGTGT 251 CGCTGGCTTC GGTGATTGCC TCTCAAATCT TCCTTTACGA AGATTTCAAC 301 CAAATGCGGA AAACCGTGGA GCTTTCTGCG GTTTTCTTGT CCAATATTTA 351 TCTGGGGTTT CAGCAGGGGT ATTTCGATTT GAGTGCCGAC GAGAACCCCG 401 TACTGCATAT CTGGTCTTTG GCAGTAGAGG AACAGTATTA CCTCCTGTAT 451 CCCCTTTTGC TGATATTTTG CTGCAAAAAA ACCAAATCGC TACGGGTGCT 501 GCGTAACATC AGCATCATCC TGTTTTTGAT TTTGACTGCC TCATCGTTTT 551 TGCCAAGCGG GTTTTATACC GACATCCTCA ACCAACCCAA TACTTATTAC 601 CTTTCGACAC TGAGGTTTCC CGAGCTGTTG GCAGGTTCGC TGCTGGCGGT 651 TTACGGGCAA ACGCAAAACG GCAGACGGCA AACAGCAAAT GGAAAACGGC 701 AGTTGCTTTC ATCACTCTGC TTCGGCGCAT TGCTTGCCTG CCTGTTCGTG 751 ATTGACAAAC ACAATCCGTT TATCCCGGGA ATGACCCTGC TCCTTCCCTG 801 CCTGCTGACG GCACTGCTTA TCCGGAGTAT GCAATACGGG ACACTTCCGA 851 CCCGCATCCT GTCGGCAAGC CCCATCGTAT TTGTCGGCAA AATCTCTTAT 901 TCCCTATACC TGTACCATTG GATTTTTATT GCTTTCGCCC ATTACATTAC 951 AGGCGACAAA CAGCTCGGAC TGCCTGCCGT ATCGGCGGTT GCCGCGTTGA 1001 CGGCCGGATT TTCCCTGTTG AGTTATTATT TGATTGAACA GCCGCTTAGA 1051 AAACGGAAGA TGACCTTCAA AAAGGCATTT TTCTGCCTCT ATCTCGCCCC 1101 GTCCCTGATA CTTGTCGGTT ACAACCTGTA CGCAAGGGGG ATATTGAAAC 1151 AGGAACACCT CCGCCCGTTG CCCGGCGCGC CCCTTGCTGC GGAAAATCAT 1201 TTTCCGGAAA CCGTCCTGAC CCTCGGCGAC TCGCACGCCG GACACCTGAG 1251 GGGGTTTCTG GATTATGTCG GCAGCCGGGA AGGGTGGAAA GCCAAAATCC 1301 TGTCCCTCGA TTCGGAGTGT TTGGTTTGGG TAGATGAGAA GCTGGCAGAC 1351 AACCCGTTAT GTCGAAAATA CCGGGATGAA GTTGAAAAAG CCGAAGCCGT 1401 TTTCATTGCC CAATTCTATG ATTTGAGGAT GGGCGGCCAG CCTGTGCCGA 1451 GATTTGAAGC GCAATCCTTC CTAATACCCG GGTTCCCAGC CCGATTCAGG 1501 GAAACCGTCA AAAGGATAGC CGCCGTCAAA CCCGTCTATG TTTTTGCAAA 1551 CAACACATCA ATCAGCCGTT CGCCCCTGAG GGAGGAAAAA TTGAAAAGAT 1601 TTGCCGCAAA CCAATATCTC CGCCCCATTC AGGCTATGGG CGACATCGGC 1651 AAGAGCAATC AGGCGGTCTT TGATTTGATT AAAGATATTC CCAATGTGCA 1701 TTGGGTGGAC GCACAAAAAT ACCTGCCCAA AAACACGGTC GAAATATACG 1751 GCCGCTATCT TTACGGCGAC CAAGACCACC TGACCTATTT CGGTTCTTAT 1801 TATATGGGGC GGGAATTCCA CAAACACGAA CGCCTGCTTA AATCTTCCCA 1851 CGGCGGCGCA TTGCAGTAG This coπesponds to the amino acid sequence <SEQ ID 830; ORF128-l>: 1 MQAVRYRPEI DGLRAVAVLS VMIFHLNNRW LPGGFLGVDI FFVISGFLIT 51 GIILSEIQNG SFSFRDFYTR RIKRIYPAFI AAVSIASVIA SQIFLYEDFN 101 QMRKTVELSA VFLSNIYLGF QQGYFDLSAD ENPVLHIWSL AVEEQYYLLY 151 PLLLIFCCKK TKSLRVLRNI SIILFLILTA SSFLPSGFYT DILNQPNTYY 201 LSTLRFPELL AGSLLAVYGQ TQNGRRQTAN GKRQLLSSLC FGALLACLFV 251 IDKHNPFIPG MTLLLPCLLT ALLIRSMQYG TLPTRILSAS PIVFVGKISY 301 SLYLYHWIFI AFAHYITGDK QLGLPAVSAV AALTAGFSLL SYYLIEQPLR 351 KRKMTFKKAF FCLYLAPSLI LVGYNLYARG ILKQEHLRPL PGAPLAAENH 401 FPETVLTLGD SHAGHLRGFL DYVGSREGWK AKILSLDSEC LVWVDEKLAD 451 NPLCRKYRDE VEKAEAVFIA QFYDLRMGGQ PVPRFEAQSF LIPGFPARFR 501 ETVKRIAAVK PVYVFANNTS ISRSPLREEK LKRFAANQYL RPIQAMGDIG 551 KSNQAVFDLI KDIPNVHWVD AQKYLPKNTV EIYGRYLYGD QDHLTYFGSY 601 YMGREFHKHE RLLKSSHGGA LQ* Computer analysis of this amino acid sequence gave the following results: Homology with hypothetical integral membrane protein HI0392 of H.influenzae (accession number U32723) ORF 128 and HI0392 show 52% aa identity in 180aa overlap: Orfl28 1 VSLASVIASQIFLYEDFNQMRKTVELSAVFLSNIYLGFQQGYFDLSADENPVLHIWSLAV 60 ++L S IAS IF+Y DFN++RKT+EL+ FLSN YLG QGYFDLSA+ENPVLHIWSLAV HI0392 46 MALVSFIASAIFIYNDFNKLRKTIELAIAFLSNFYLGLTQGYFDLSANENPVLHIWSLAV 105 Orfl28 61 EEQXXXXXXXXXIFCCKKTKSLRVLRNISIILFLILTASSFLPSGFYTDILNQPNTYYLS 120 E Q I KK + ++VL I++ILF IL A+SF+ + FY ++L+QPN YYLS HI0392 106 EGQYYLIFPLILILAYKKFREVKVLFIITLILFFILLATSFVSANFYKEVLHQPNIYYLS 165 0rfl28 121 TLRFPELLAGSLLAVYGQTQNGRRQTANGKRQLLSSLCFGALLACLFVIDKHNPFIPGMT 180 LRFPELL GSLLA+Y N + Q + +L+ L L +CLF+++ + FIPG+T HI0392 166 NLRFPELLVGSLLAIYHNLSN-KVQLSKQVNNILAILSTLLLFSCLFLMNNNIAFIPGIT 224 Homology with a predicted ORF from N.meninεitidis (sfrain Al ORF128 shows 98.0% identity over a 244aa overlap with an ORF (ORF128a) from strain A of N. meningitidis: 10 20 30 orf128.pep VSLASVIASQIFLYEDFNQMRKTVELSAVF II I I I II II I I I I I I II I I II I I I II I I I I orf128a ILSEIQNGSFSFRDFYTRRIKRIYPAFIAAVSLASVIASQIFLYEDFNQMRKTVELSAVF 60 70 80 90 100 110 40 50 60 70 80 90 orf128. pep LSNIYLGFQQGYFDLSADENPVLHIWSLAVEEQYYLLYPLLLIFCCKKTKSLRVLRNISI I II I II II I I I I I II I I II II I I I II I I I I I II II I II I I II II II I I I I I II II I I I I I orf128a LSNIYLGFQQGYFDLSADENPVLHIWSLAVEEQYYLLYPLLLIFCCKKTKSLRVLRNISI 120 130 140 150 160 170 100 110 120 130 140 150 orf128. pep ILFLILTASSFLPSGFYTDILNQPNTYYLSTLRFPELLAGSLLAVYGQTQNGRRQTANGK I II I II I I : I I I I II I II I II I I I II I I I I I II I I I I I I II I II II I I I I I II II I I I I I orf128a ILFLILTATSFLPSGFYTDILNQPNTYYLSTLRFPELLAGSLLAVYGQTQNGRRQTANGK 180 190 200 210 220 230 160 170 180 190 200 210 orf128. pep RQLLSSLCFGALLACLFVIDKHNPFIPGMTLLLPCLLTALLIRSMQYGTLPTRILSASPI I I I I I I II I I I I II I I I I I II II I II I I I I I I I I I I II II II I II II I I I I II II I I I I I orf128a RQLLSSLCFGALLACLFVIDKHNPFIPGMTLLLPCLLTALLIRSMQYGTLPTRILSASPI 240 250 260 270 280 290 220 230 240 orf128.pep VFVGKISYSLYLYHWIFIAFAPLIRGGKQLGLPA II I II I I I II II II I II I I II I I I I I II I I orf128a VFVGKISYSLYLYHWIFIAFAHYITGDKQLGLPAVSAVAALTAGFSLLSYYLIEQPLRKR 300 310 320 330 340 350 orf128a KMTFKKAFFCLYLAPSLILVGYNLYARGILKQEHLRPLPGAPLAAENHFPETVLTLGDSH 360 370 380 390 400 410 The complete length ORF128a nucleotide sequence <SEQ LD 831> is: 1 ATGCAAGCTG TCCGATACAG ACCGGAAATT GACGGATTGC GGGCCGTCGC 51 CGTGCTATCC GTCATGATTT TCCACCTGAA TAACCGCTGG CTGCCCGGAG 101 GATTCCTGGG GGTGGACATT TTCTTTGTCA TCTCAGGATT CCTCATTACC 151 GGCATCATTC TTTCTGAAAT ACAGAACGGT TCTTTTTCTT TCCGGGATTT 201 TTATACCCGC AGGATTAAGC GGATTTATCC TGCTTTTATT GCGGCCGTGT 251 CGCTGGCTTC GGTGATTGCC TCTCAAATCT TCCTTTACGA AGATTTCAAC 301 CAAATGCGGA AAACCGTGGA GCTTTCTGCG GTTTTCTTGT CCAATATTTA 351 TCTGGGGTTT CAGCAGGGGT ATTTCGATTT GAGTGCCGAC GAGAACCCCG 401 TACTGCATAT CTGGTCTTTG GCAGTAGAGG AACAGTATTA CCTCCTGTAT 451 CCTCTTTTGC TGATATTTTG CTGCAAAAAA ACAAAATCGC TACGGGTGCT 501 GCGTAACATC AGCATCATCC TATTTCTGAT TTTGACTGCC ACATCGTTTT 551 TGCCAAGCGG GTTTTATACC GATATTCTCA ACCAACCCAA TACTTATTAC 601 CTTTCGACAC TGAGGTTTCC CGAGCTGTTG GCAGGTTCGC TGCTGGCGGT 651 TTACGGGCAA ACGCAAAACG GCAGACGGCA AACAGCAAAT GGAAAACGGC 701 AGTTGCTTTC ATCACTCTGC TTCGGCGCAT TGCTTGCCTG CCTGTTCGTG 751 ATTGACAAAC ACAATCCGTT TATCCCGGGA ATGACCCTGC TCCTTCCCTG 801 CCTGCTGACG GCACTGCTTA TCCGGAGTAT GCAATACGGG ACACTTCCGA 851 CCCGCATCCT GTCGGCAAGC CCCATCGTAT TTGTCGGCAA AATCTCTTAT 901 TCCCTATACC TGTACCATTG GATTTTTATT GCTTTCGCCC ATTACATTAC 951 AGGCGACAAA CAGCTCGGAC TGCCTGCCGT ATCGGCGGTT GCCGCGTTGA 1001 CGGCCGGATT TTCCCTGTTG AGTTATTATT TGATTGAACA GCCGCTTAGA 1051 AAACGGAAGA TGACCTTCAA AAAGGCATTT TTCTGCCTCT ATCTCGCCCC 1101 GTCCCTGATA CTTGTCGGTT ACAACCTGTA CGCAAGGGGG ATATTGAAAC 1151 AGGAACACCT CCGCCCGTTG CCCGGCGCGC CCCTTGCTGC GGAAAATCAT 1201 TTTCCGGAAA CCGTCCTGAC CCTCGGCGAC TCGCACGCCG GACACCTGCG 1251 GGGGTTTCTG GATTATGTCG GCAGCCGGGA AGGGTGGAAA GCCAAAATCC 1301 TGTCCCTCGA TTCGGAGTGT TTGGTTTGGG TAGATGAGAA GCTGGCAGAC 1351 AACCCGTTAT GTCGAAAATA CCGGGATGAA GTTGAAAAAG CCGAAGCCGT 1401 TTTCATTGCC CAATTCTATG ATTTGAGGAT GGGCGGCCAG CCCGTGCCGA 1451 GATTTGAAGC GCAATCCTTC CTAATACCCG GGTTCCCAGC CCGATTCAGG 1501 GAAACCGTCA AAAGGATAGC CGCCGTCAAA CCCGTCTATG TTTTTGCAAA 1551 CAACACATCA ATCAGCCGTT CGCCCCTGAG GGAGGAAAAA TTGAAAAGAT 1601 TTGCCGCAAA CCAATATCTC CGCCCCATTC AGGCTATGGG CGACATCGGC 1651 AAGAGCAATC AGGCGGTCTT TGATTTGATT AAAGATATTC CCAATGTGCA 1701 TTGGGTGGAC GCACAAAAAT ACCTGCCCAA AAACACGGTC GAAATATACG 1751 GCCGCTATCT TTACGGCGAC CAAGACCACC TGACCTATTT CGGTTCTTAT 1801 TATATGGGGC GGGAATTTCA CAAACACGAA CGCCTGCTTA AATCTTCTCG 1851 CGACGGCGCA TTGCAGTAG This encodes a protein having amino acid sequence <SEQ LD 832>: 1 MQAVRYRPEI_ DGLRAVAVLS VMIFHLNNRW LPGGFLGVDI FFVISGFLIT 51 GIILSEIQNG SFSFRDFYTR RIKRIYPAFI AAVSLASVIA SQIFLYEDFN 101 QMRKTVELSA VFLSNIYLGF QQGYFDLSAD ENPVLHIWSL AVEEQYYLLY 151 PLLLIFCCKK TKSLRVLRNI_ SIILFLILTA TSFLPSGFYT DILNQPNTYY 201 LSTLRFPELL AGSLLAVYGQ TQNGRRQTAN GKRQLLSSLC FGALLACLFV 251 IDKHNPFIPG MTLLLPCLLT ALLIRSMQYG TLPTRILSAS PIVFVGKISY 301 SLYLYHWIFI AFAHYITGDK QLGLPAVSAV AALTAGFSLL SYYLIEQPLR 351 KRKMTFKKAF FCLYLAPSLI LVGYNLYARG ILKQEHLRPL PGAPLAAENH 401 FPETVLTLGD SHAGHLRGFL DYVGSREGWK AKILSLDSEC LVWVDEKLAD 451 NPLCRKYRDE VEKAEAVFIA QFYDLRMGGQ PVPRFEAQSF LIPGFPARFR 501 ETVKRIAAVK PVYVFANNTS ISRSPLREEK LKRFAANQYL RPIQAMGDIG 551 KSNQAVFDLI KDIPNVHWVD AQKYLPKNTV EIYGRYLYGD QDHLTYFGSY 601 YMGREFHKHE RLLKSSRDGA LQ* ORF128a and ORF128-1 show 99.5% identity in 622 aa overlap: orfl28a .pep MQAVRYRPEIDGLRAVAVLSVMIFHLNNRWLPGGFLGVDIFFVISGFLITGIILSEIQNG I I I I II I II I I I I II I I I I II I II I II II II II II I II I II I I I I I I II I I II I I I I I II orfl28- 1 MQAVRYRPEIDGLRAVAVLSVMIFHLNNRWLPGGFLGVDIFFVISGFLITGIILSEIQNG orfl28a .pep SFSFRDFYTRRIKRIYPAFIAAVSLASVIASQIFLYEDFNQMRKTVELSAVFLSNIYLGF I I II II I II I I I I II II I I I I I I II II I II II I I I II I I I I I II I I I I I I II II II II I I orfl28- 1 SFSFRDFYTRRIKRIYPAFIAAVSLASVIASQIFLYEDFNQMRKTVELSAVFLSNIYLGF orfl28a .pep QQGYFDLSADENPVLHIWSLAVEEQYYLLYPLLLIFCCKKTKSLRVLRNISIILFLILTA II I II II I II I I II I I I I II I I I I II I I I I I II I I I I I I I I I I I I II I I I I I II II I I I I orfl28- 1 QQGYFDLSADENPVLHIWSLAVEEQYYLLYPLLLIFCCKKTKSLRVLRNISIILFLILTA orfl28a .pep TSFLPSGFYTDILNQPNTYYLSTLRFPELLAGSLLAVYGQTQNGRRQTANGKRQLLSSLC : I I I I II I I I I I I I II I I I II I II I II I I I II II II I I II I I I I I I I I I II II I I I I I II orfl28- 1 SSFLPSGFYTDILNQPNTYYLSTLRFPELLAGSLLAVYGQTQNGRRQTANGKRQLLSSLC orfl28a .pep FGALLACLFVIDKHNPFIPGMTLLLPCLLTALLIRSMQYGTLPTRILSASPIVFVGKISY I I I I I II I I II I I I II I I I II I II I I II II II II II I II I II I I I II I I I I I II II II I I orfl28- 1 FGALLACLFVIDKHNPFIPGMTLLLPCLLTALLIRSMQYGTLPTRILSASPIVFVGKISY orfl28a • pep SLYLYHWIFIAFAHYITGDKQLGLPAVSAVAALTAGFSLLSYYLIEQPLRKRKMTFKKAF II I I I I II I I I I I II II II I II I I II I I I II II II I II I I I I I I II I I I I I II I I I I I II orfl28- 1 SLYLYHWIFIAFAHYITGDKQLGLPAVSAVAALTAGFSLLSYYLIEQPLRKRKMTFKKAF orfl28a •pep FCLYLAPSLILVGYNLYARGILKQEHLRPLPGAPLAAENHFPETVLTLGDSHAGHLRGFL I II I II I II I I II I I I I I II I I I I I II II I I I I I II I I II I I I II I I I II I II I I I I I I I orfl28- 1 FCLYLAPSLILVGYNLYARGILKQEHLRPLPGAPLAAENHFPETVLTLGDSHAGHLRGFL orfl28a .pep DYVGSREGWKAKILSLDSECLVWVDEKLADNPLCRKYRDEVEKAEAVFIAQFYDLRMGGQ I I II I II I II I I II II I I I II I I I I I II II I I I I I I II I II I II I I I I I I II I I II II I I orfl28- 1 DYVGSREGWKAKILSLDSECLVWVDEKLADNPLCRKYRDEVEKAEAVFIAQFYDLRMGGQ orfl28a • pep PVPRFEAQSFLIPGFPARFRETVKRIAAVKPVYVFANNTSISRSPLREEKLKRFAANQYL II I II II I I II I I I I II II I II I II I I I I II I I I I II I II I II I I I I I I I I II I I I I II I orfl28- 1 PVPRFEAQSFLIPGFPARFRETVKRIAAVKPVYVFANNTSISRSPLREEKLKRFAANQYL orfl28a •pep RPIQAMGDIGKSNQAVFDLIKDIPNVHWVDAQKYLPKNTVEIYGRYLYGDQDHLTYFGSY I I I I I II I I II I I I I I I II I II I I II I I I II I I I I II I II I I I I I I I I II I II I I I I I II orfl28- 1 RPIQAMGDIGKSNQAVFDLIKDIPNVHWVDAQKYLPKNTVEIYGRYLYGDQDHLTYFGSY orfl28a •pep YMGREFHKHERLLKSSRDGALQX I I II II I I II I I I I II : I I II I orfl28- 1 YMGREFHKHERLLKSSHGGALQX Homology with a predicted ORF from Nεonorrhoeae ORF 128 shows 93.4% identity over 244 aa overlap with a predicted ORF (ORF128ng) from N. gonorrhoeae: orf128. pep VSLASVIASQIFLYEDFNQMRKTVELSAVF 30 II I II I I I I I II I II I II I I I II: I I I: I I orfl28ng ILSEIQNGSFSFRDFYTRRIKRIYPAFIAAVSLASVIASQIFLYEDFNQMRKTIELSTVF 112 orf128. pep LSNIYLGFQQGYFDLSADENPVLHIWSLAVEEQYYLLYPLLLIFCCKKTKSLRVLRNISI 90 I I I I II I I : II I I I I I II II I I I I I I II I II I II I I II I II I I I I I I I II I II I I I I I orfl28ng LSNIYLGFRLGYFDLSADENPVLHIWSLAVEEQYYLLYPLLLIFCYKKTKSLRVLRNISI 172 orfl28.pep ILFLILTASSFLPSGFYTDILNQPNTYYLSTLRFPELLAGSLLAVYGQTQNGRRQTANGK 150 I I II I I I I I I I I II II I I I I I II I II I II II I I II I: I I I I II I II I I I II II I III orfl28ng ILFLILTASSFLPAGFYTDILNQPNTYYLSTLRFPELLVGSLLAVYGQTQNGRRQTENGK 232 orf128.pep RQLLSSLCFGALLACLFVIDKHNPFIPGMTLLLPCLLTALLIRSMQYGTLPTRILSASPI 210 II I I I I I II I I I M I I I II II : I M I I: I I I I I II I I II I I I I I I II I I I II II I I I I I orfl28ng RQLLSLLCFGALLVCLFVIDKHDPFIPGITLLLPCLLTALLIRSMQYGTLPTRILSASPI 292 orfl28.pep VFVGKISYSLYLYHWIFIAFAPLIRGGKQLGLPA 244 II II I II II I I I I I I I II II I I I I II II II orfl28ng VFVGKISYSLYLYHWIFIAFAHYITGDKQLGLPAVSAVAALTAGFSLLSYYLIEQPLRKR 352 The complete length ORF128ng nucleotide sequence <SEQ ID 833> is: 1 ATGCAAGCTG TCCGATACAG GCCTGAAATT GACGGATTGC GGGCCGTCGC 51 CGTGCTATCC GTCATTATTT TCCACCTGAA TAACCGCTGG CTGCCCGGAG 101 GATTCCTGGG GGTGGACATT TTCTTTGTCA TCTCGGGATT CCTCATTACC 151 AACATCATTC TTTCTGAAAT ACAGAACGGT TCTTTTTCTT TCCGGGATTT 201 TTATACCCGC AGGATTAAGC GGATTTATCC TGCTTTTATT GCGGCCGTGT 251 CCCTGGCTTC GGTGATTGCT TCTCAAATCT TCCTTTACGA AGATTTCAAC 301 CAAATGAGGA AAACCATAGA GCTTTCTACG GTTTTTTTGT CCAATATTTA 351 TTTGGGGTTC CGATTGGGGT ATTTCGATTT GAGTGCCGAC GAGAACCCCG 401 TACTGCATAT CTGGTCTTTG GCGGTAGAGG AACAGTATTA CCTCCTGTAT 451 CCTCTTTTGC TGATATTCTG TTACAAAAAA ACCAAATCAC TACGGGTGCT 501 GCGTAATATC AGCATCATCC TGTTTCTGAT TTTGACCGCA TCATCGTTTT 551 TGCCGGCCGG GTTTTATACC GACATCCTCA ACCAACCcaa TACTTATTAC 601 CTTTCGACAC TGAGGTTTCC CGAGCTGTTG GTGGGTTCGC TGTTGGCGGT 651 TTACGGGCAA ACGCAAAACG GCAGACGGCA AACAGAAAAT GGAAAACGGC 701 AGTTGCTTTC ATTACTCTGT TTCGGCGCat tgCTTGTCTG CCTGTTCGTG 751 ATCGACAAAC ACGATCCGTT TATCCCGGGA ATAACCCTGC TCCTTCCCTG 801 CCTGCTGACG GCGCTGCTTA TCCGGAGTAT GCAATACGGG ACACTTCCGA 851 CCCGCATCCT GTCGGCAAGC CCCATCGTAT TTGTCGGCAA AATCTCTTAT 901 TCCCTATACC TGTACCATTG GATTTTTATT GCCTTCGCCC ATTACATTAC 951 AGGCGACAAA CAGCTCGGAC TGCCTGCCGT ATCGGCGGTT GCCGCGTTGA 1001 CGGCCGGATT TTCCCTGTTG AGCTATTATT TGATTGAACA GCCGCTTAGA 1051 AAACGGAAGA TGACCTTCAA AAAGGCATTT TTCTGCCTTT ATCTCGCCCC 1101 GTCCCTGATG CTTGTCGGTT ACAACCTGTA TTCAAGAGGG ATATTGAAAC 1151 AGGAACACCT CCGCCCGCTG CCCGGCACGC CCGTTGCTGC GGAAAATAAT 1201 TTTCCGGAAA CCGTCTTGAC CCTCGGCGAC TCGCACGCCG GACACCTGCG 1251 GGGGTTTCTG GATTATGTCG GCGGCAGGGA AGGGTGGAAA GCTAAAATCC 1301 TGTCCCTCGA TTCGGAGTGT TTGGTTTGGG TGGATGAGAA GCTGGCAGAC 1351 AACCCGTTGT GCCGAAAATA CCGGGATGAA GTTGAAAAAG CCGAAGCTGT 1401 TTTCATTGCC CAATTCTATG ATTTGAGGAT GGGCGGCCAG CCCGTGCCGA 1451 GATTTGAAGC GCAATCCTTC CTGATACCCG GGTTCAAAGC CCGATTCAGG 1501 GAAACCGTCA AGAGGATAGC CGCCGTCAAA CCTGTATATG TTTTTGCAAA 1551 CAATACATCA ATCAGCCGTT CTCCCTTGAG GGAGGAAAAA TTGAAAAGAT 1601 TTGCTATAAA CCAATACCTC CGGCCTATTC GGGCTATGGG CGACATCGGC 1651 AAGAGCAATC AGGCGGTCTT TGATTTGGTT AAAGATATTC CCAATGTGCA 1701 TTGGGTGGAC GCACAAAAAT ACCTGCCCAA AAACACGGTC GAAATACACG 1751 GACGCTATCT TTACGGCGAC CAAGACCACC TGACCTATTT CGGTTCTTAT 1801 TATATGGGGC GGGAATTTCA CAAACACGAA CGCCTGCTCA AGCATTCCCG 1851 AGGCGGCGCA TTGCAGTAG This encodes a protein having amino acid sequence <SEQ ED 834>: 1 MQAVRYRPEI DGLRAVAVLS VIIFHLNNRW LPGGFLGVDI FFVISGFLIT 51 NIILSEIQNG SFSFRDFYTR RIKRIYPAFI AAVSLASVIA SQIFLYEDFN 101 QMRKTIELST VFLSNIYLGF RLGYFDLSAD ENPVLHIWSL AVEEQYYLLY 151 PLLLIFCYKK TKSLRVLRNI SIILFLILTA SSFLPAGFYT DILNQPNTYY 201 LSTLRFPELL VGSLLAVYGQ TQNGRRQTEN GKRQLLSLLC FGALLVCLFV 251 IDKHDPFIPG ITLLLPCLLT ALLIRSMQYG TLPTRILSAS PIVFVGKISY 301 SLYLYHWIFI AFAHYITGDK QLGLPAVSAV AALTAGFSLL SYYLIEQPLR 351 KRKMTFKKAF FCLYLAPSLM LVGYNLYSRG ILKQEHLRPL PGTPVAAENN 401 FPETVLTLGD SHAGHLRGFL DYVGGREGWK AKILSLDSEC LVWVDEKLAD 451 NPLCRKYRDE VEKAEAVFIA QFYDLRMGGQ PVPRFEAQSF LIPGFKARFR 501 ETVKRIAAVK PVYVFANNTS ISRSPLREEK LKRFAINQYL RPIRAMGDIG 551 KSNQAVFDLV KDIPNVHWVD AQKYLPKNTV EIHGRYLYGD QDHLTYFGSY 601 YMGREFHKHE RLLKHSRGGA LQ* ORF128ng and ORF128-1 show 95.7% identity in 622 aa overlap: orf128-1. pep MQAVRYRPEIDGLRAVAVLSVMIFHLNNRWLPGGFLGVDIFFVISGFLITGIILSEIQNG I II I I II I I I I II I II II II I : II I I II I II I II I I I I I I I I I I I I II I I : I II I I I I I I orfl28ng MQAVRYRPEIDGLRAVAVLSVIIFHLNNRWLPGGFLGVDIFFVISGFLITNIILSEIQNG orf128-1. pep SFSFRDFYTRRIKRIYPAFIAAVSLASVIASQIFLYEDFNQMRKTVELSAVFLSNIYLGF II I I I I I II II I II II I I I II I I II I I II I I I I II I I I I I I I I I I: I I I : II I II II II I orfl28ng SFSFRDFYTRRIKRIYPAFIAAVSLASVIASQIFLYEDFNQMRKTIELSTVFLSNIYLGF orf128-1. pep QQGYFDLSADENPVLHIWSLAVEEQYYLLYPLLLIFCCKKTKSLRVLRNISIILFLILTA : I II II I I I I I I I I I I I I I I I I II II I I I II I I I II I I II II I I I I I II II I I I II II orfl28ng RLGYFDLSADENPVLHIWSLAVEEQYYLLYPLLLIFCYKKTKSLRVLRNISIILFLILTA orf128-1. pep SSFLPSGFYTDILNQPNTYYLSTLRFPELLAGSLLAVYGQTQNGRRQTANGKRQLLSSLC II I II: II II I I I II II I I II I I II I I I I I: I II II I I I I I II I I I I I I II I I I I I II orfl28ng SSFLPAGFYTDILNQPNTYYLSTLRFPELLVGSLLAVYGQTQNGRRQTENGKRQLLSLLC orf128-1. pep FGALLACLFVIDKHNPFIPGMTLLLPCLLTALLIRSMQYGTLPTRILSASPIVFVGKISY I I II I : I II I I I II : I I I II : I I II II I I I I I II II I I II I II I I I I II I II I I I I I I I I orfl28ng FGALLVCLFVIDKHDPFIPGITLLLPCLLTALLIRSMQYGTLPTRILSASPIVFVGKISY orf128-1. pep SLYLYHWIFIAFAHYITGDKQLGLPAVSAVAALTAGFSLLSYYLIEQPLRKRKMTFKKAF I I II I I I II I II II II I I I I I II I I II II I I I II II I I II I II I I II I I I II I I I I I I I I orfl28ng SLYLYHWIFIAFAHYITGDKQLGLPAVSAVAALTAGFSLLSYYLIEQPLRKRKMTFKKAF orf128-1. pep FCLYLAPSLILVGYNLYARGILKQEHLRPLPGAPLAAENHFPETVLTLGDSHAGHLRGFL I I I I I I I II: II II II I : I II I I II I I II I I I : I : I II I : I I I I I I I I I II II I I II II I orfl28ng FCLYLAPSLMLVGYNLYSRGILKQEHLRPLPGTPVAAENNFPETVLTLGDSHAGHLRGFL orf128-1. pep DYVGSREGWKAKILSLDSECLVWVDEKLADNPLCRKYRDEVEKAEAVFIAQFYDLRMGGQ I II I: I I I I II I I I II II II I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I II II II II I orfl28ng DYVGGREGWKAKILSLDSECLVWVDEKLADNPLCRKYRDEVEKAEAVFIAQFYDLRMGGQ orf128-1. pep PVPRFEAQSFLIPGFPARFRETVKRIAAVKPVYVFANNTSISRSPLREEKLKRFAANQYL I II I I I I I I II I II I II I II I I I I I I II II II I I II I I I II I I II I I II I I II I I II I orfl28ng PVPRFEAQSFLIPGFKARFRETVKRIAAVKPVYVFANNTSISRSPLREEKLKRFAINQYL orf128-1.pep RPIQAMGDIGKSNQAVFDLIKDIPNVHWVDAQKYLPKNTVEIYGRYLYGDQDHLTYFGSY I I I : I I I II I II II II I I I : I I II I I I I II I I I I I I I I I I I I : I I I I I I I II I I I I I I I I orfl28ng RPIRAMGDIGKSNQAVFDLVKDIPNVHWVDAQKYLPKNTVEIHGRYLYGDQDHLTYFGSY orf128-1. pep YMGREFHKHERLLKSSHGGALQX I II I II I II II I II I : I I II I I orfl28ng YMGREFHKHERLLKHSRGGALQX 610 620 En addition, ORF218ng shows homology to a hypothetical H.influenzae protein: sp|P43993|Y392_HAEIN HYPOTHETICAL PROTEIN HI0392 >gi | 1074385 Ipir | IB64007 hypothetical protein HI0392 - Haemophilus influenzae (strain Rd KW20) >gi 11573364 (U32723) H. influenzae predicted coding region HI0392 [Haemophilus influenzae] Length = 245 Score = 239 bits (604), Expect = 3e-62 Identities = 124/225 (55%), Positives = 152/225 (67%), Gaps = 1/225 (0%) Query: 38 VDIFFVISGFLITNIILSEIQNGSFSFRDFYTRRIKRIYPXXXXXXXXXXXXXXXXFLYE 97 +DIFFVISGFLIT II++EIQ SFS + FYTRRIKRIYP F+Y Sbjct: 1 MDIFFVISGFLITGIIITEIQQNSFSLKQFYTRRIKRIYPAFITVMALVSFIASAIFIYN 60 Query: 98 DFNQMRKTIELSTVFLSNIYLGFRLGYFDLSADENPVLHIWSLAVEEQXXXXXXXXXIFC 157 DFN++RKTIEL+ FLSN YLG GYFDLSA+ENPVLHIWSLAVE Q I Sbjct: 61 DFNKLRKTIELAIAFLSNFYLGLTQGYFDLSANENPVLHIWSLAVEGQYYLIFPLILILA 120 Query: 158 YKKTKSLRVLRNISIILFLILTASSFLPAGFYTDILNQPNTYYLSTLRFPELLVGSLLAV 217 YKK + ++VL I++ILF IL A+SF+ A FY ++L+QPN YYLS LRFPELLVGSLLA+ Sbjct: 121 YKKFREVKVLFIITLILFFILLATSFVSANFYKEVLHQPNIYYLSNLRFPELLVGSLLAI 180 Query: 218 YGQTQNGRRQTENGKRQLLSLLCFGALLVCLFVIDKHDPFIPGIT 262 Y N + Q +L++L L CLF+++ + FIPGIT Sbjct: 181 YHNLSN-KVQLSKQVNNILAILSTLLLFSCLFLMNNNIAFIPGIT 224 This analysis, including the identification of several putative transmembrane domains, suggests that these proteins from N. meningitidis and N.gonorrhoeae, and their epitopes, could be useful .antigens for vaccines or diagnostics, or for raising antibodies. Example 99 The following partial DNA sequence was identified in N. meningitidis <SEQ ED 835>: 1 ..ATTATTTACG AATACCGCTG GATGTTTCTT TACGGCGCAC TGACGACCTT 51 GGGGCTGACG GTCGTGGCAA C.GCGGGCGG TTCGGTATTG GGTCTGTTGT 101 TGGCGTTGGC GCGCCTGATT CACTTGGAAA AAGCCGGTGC GCCGATGCGC 151 GTGCTGGCGT GGGCGTTGCG TAAAGTTTCG CTGCTGTATG TTACGCTGTT 201 CCGGGGTACG CCGCTGTTTG TGCAGATTGT GATTTGGGCG TATGTGTGGT 251 TTCCGTTTTT CGTC .. This coπesponds to the amino acid sequence <SEQ LD 836; ORF129>: 1 .. IIΪΕYRWMFL YGALTTLGLT WAXAGGSVL GLLLALARLI HLEKAGAPMR 51 VLAWALRKVS LLYVTLFRGT PLFVQIVIWA YVWFPFFV.. Further work revealed the complete nucleotide sequence <SEQ ID 837>: 1 ATGGATTTTC GTTTTGACAT TATTTACGAA TACCGCTGGA TGTTTCTTTA 51 CGGCGCACTG ACGACCTTGG GGCTGACGGT CGTGGCAACG GCGGGCGGTT 101 CGGTATTGGG TCTGTTGTTG GCGTTGGCGC GCCTGATTCA CTTGGAAAAA 151 GCCGGTGCGC CGATGCGCGT GCTGGCGTGG GCGTTGCGTA AAGTTTCGCT 201 GCTGTATGTT ACGCTGTTCC GGGGTACGCC GCTGTTTGTG CAGATTGTGA 251 TTTGGGCGTA TGTGTGGTTT CCGTTTTTCG TCCATCCTTC AGACGGCATT 301 TTGGTCAGCG GCGAGGCGGC AATCGCGCTG CGTCGCGGAT ACGGGCCGCT 351 GATTGCCGGT TCTTTGGCAC TGATCGCCAA CTCGGGGGCG TATATCTGTG 401 AGATTTTCCG CGCGGGCATC CAGTCTATAG ACAAAGGACA GATGGAGGCG 451 GCGCGTTCTT TGGGGCTGAC CTATCCGCAG GCGATGCGCT ATGTGATTCT 501 GCCGCAGGCA TTGCGCCGCA TGCTGCCGCC TTTGGCGAGC GAGTTCATCA 551 CGCTCTTGAA AGACAGCTCG CTGCTGTCGG TCATTGCTGT GGCGGAGTTG 601 GCGTATGTTC AGAATACGAT TACGGGCCGG TATTCGGTTT ATGAAGAACC 651 GCTTTACACC GTCGCCCTGA TTTATCTGTT GATGACGACT TTCTTAGGCT 701 GGATATTCCT GCGTTTGGAA AAACGTTACA ATCCGCAACA CCGCTGA This coπesponds to the amino acid sequence <SEQ ID 838; ORF129-l>: 1 MDFRFDIIiΕ YRWMFLYGAL TTLGLTWAT AGGSVLGLLL ALARLIHLEK 51 AGAPMRVLAW ALRKVSLLYV TLFRGTPLFV QIVIWAYVWF PFFVHPSDGI 101 LVSGEAAIAL RRGYGPLIAG SLALIANSGA YICEIFRAGI QSIDKGQMEA 151 ARSLGLTYPQ AMRYVILPQA LRRMLPPLAS EFITLLKDSS LLSVIAVAEL 201 AYVQNTITGR YSVYEEPLYT VALIYLLMTT FLGWIFLRLE KRYNPQHR* Computer analysis of this amino acid sequence gave the following results: Homology with a predicted ORF from N.meninsitidis (sfrain A) ORF129 shows 98.9% identity over a 88aa overlap with an ORF (ORF129a) from strain A of N. meningitidis: 10 20 30 40 50 orf129.pep IIYEYRWMFLYGALTTLGLTWAXAGGSVLGLLLALARLIHLEKAGAPMRVLAW I I II I I II I II I I I II I I II I II : I I I I I I II II I I I II I II I I I II I I I II I I orf129a MDFRFDIIYEYRWMFLYGALTTLGLTWATAGGSVLGLLLALARLIHLEKAGAPMRVLAW 10 20 30 40 50 60 60 70 80 orf129. pep ALRKVSLLYVTLFRGTPLFVQIVIWAYVWFPFFV I II II I II I II I I II I II I I I II I I I I I II I II I orf129a ALRKVSLLYVTLFRGTPLFVQIVIWAYVWFPFFVHPSDGILVSGEAAIALRRGYGPLIAG 70 80 90 100 110 120 orfl29a SLALIANSGAYICEIFRAGIQSIDKGQMEAARSLGLTYPQAMRYVILPQALRRMLPPLAS 130 140 150 160 170 180 The complete length ORF129a nucleotide sequence <SEQ ID 839> is: 1 ATGGATTTTC GTTTTGACAT TATTTACGAA TACCGCTGGA TGTTTCTTTA 51 CGGCGCACTG ACGACCTTGG GGCTGACGGT CGTGGCGACG GCGGGCGGTT 101 CGGTATTGGG TCTGTTGTTG GCGTTGGCGC GCCTGATTCA CTTGGAAAAA 151 GCCGGTGCGC CGATGCGCGT GCTGGCGTGG GCGTTGCGTA AGGTTTCGCT 201 GCTGTATGTT ACGCTGTTCC GGGGTACGCC GCTGTTTGTG CAGATTGTGA 251 TTTGGGCGTA TGTGTGGTTT CCGTTTTTCG TCCATCCTTC AGACGGCATT 301 TTGGTTAGCG GCGAGGCGGC AATCGCGCTG CGTCGCGGAT ACGGGCCGCT 351 GATTGCCGGT TCTTTGGCAC TGATCGCCAA CTCGGGGGCG TATATCTGTG 401 AGATTTTCCG CGCGGGCATC CAGTCTATAG ACAAAGGACA GATGGAGGCG 451 GCGCGTTCTT TGGGGCTGAC CTATCCGCAG GCGATGCGCT ATGTGATTCT 501 GCCGCAGGCA TTGCGCCGTA TGCTGCCGCC TTTGGCGAGC GAGTTCATCA 555511 CGCTCTTGAA AGACAGCTCG CTGCTGTCGG TCATTGCTGT GGCGGAGTTG 601 GCGTATGTTC AGAATACGAT TACGGGCCGG TATTCGGTTT ATGAAGAACC 651 GCTTTACACC GTCGCCCTGA TTTATCTGTT GATGACGACT TTCTTAGGCT 701 GGATATTCCT GCGTTTGGAA AAACGTTACA ATCCGCAACA CCGCTGA This encodes a protein having amino acid sequence <SEQ ID 840>: 1 MDFRFDIIYE YRWMFLYGAL TTLGLTVVAT AGGSVLGLLL ALARLIHLEK 51 AGAPMRVLAW ALRKVSLLYV TLFRGTPLFV QIVIWAYVWF PFFVHPSDGI 101 LVSGEAAIAL RRGYGPLIAG SLALIANSGA YICEIFRAGI QSIDKGQMEA 151 ARSLGLTYPQ AMRYVILPQA LRRMLPPLAS EFITLLKDSS LLSVIAVAEL 201 AYVQNTITGR YSVYEEPLYT VALIYLLMTT FLGWIFLRLE KRYNPQHR* ORF129a and ORF129-1 show 100.0% identity in 248 aa overlap: orf129a . pep MDFRFDIIYEYRWMFLYGALTTLGLTWATAGGSVLGLLLALARLIHLEKAGAPMRVLAW I I I I II I I I I I I I I I I II II I II II I II II I I I I I I I II II I I I I I II II I II II II II I orf129-1 MDFRFDIIYEYRWMFLYGALTTLGLTWATAGGSVLGLLLALARLIHLEKAGAPMRVLAW orf129a . pep ALRKVSLLYVTLFRGTPLFVQIVIWAYVWFPFFVHPSDGILVSGEAAIALRRGYGPLIAG II I I II I I II I I I I I I II I I I II I I I I I I I II II I II I II II I I II I I I I II I I I I I I I I orf129-1 ALRKVSLLYVTLFRGTPLFVQIVIWAYVWFPFFVHPSDGILVSGEAAIALRRGYGPLIAG orf129a. pep SLALIANSGAYICEIFRAGIQSIDKGQMEAARSLGLTYPQAMRYVILPQALRRMLPPLAS I I I II I I II II I II I I I I I I II I I I I II II I I I I I II I I II II I II II I I I II I I II II I orf129-1 SLALIANSGAYICEIFRAGIQSIDKGQMEAARSLGLTYPQAMRYVILPQALRRMLPPLAS orf129a. pep EFITLLKDSSLLSVIAVAELAYVQNTITGRYSVYEEPLYTVALIYLLMTTFLGWIFLRLE I II I I I II I I I II II II I I I I I I I I I I II I I I I I I I I II I I I I I I I I I II I II I II II I I orf129-1 EFITLLKDSSLLSVIAVAELAYVQNTITGRYSVYEEPLYTVALIYLLMTTFLGWIFLRLE orf129a. pep KRYNPQHRX I I I I II I I I orf129-1 KRYNPQHRX Homology with a predicted ORF from N.gonorrhoeae ORF129 shows 98.9% identity over a 88 aa overlap with a predicted ORF (ORF129ng) from N.gonorrhoeae: orf 129 . pep IIYEYRWMFLYGALTTLGLTWAXAGGSVLGLLLALARLIHLEKAGAPMRVLAW 54 I I I I I I I I I I I I I I I I I I I I I I I : I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I orfl29ng MDFRFDIIYEYRWMFLYGALTTLGLTWATAGGSVLGLLLALARLIHLEKAGAPMRVLAW 60 orf 129 . pep ALRKVSLLYVTLFRGTPLFVQIVIWAYVWFPFFV 88 I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I orfl29ng ALRKVSLLYVTLFRGTPLFVQIVIWAYVWFPFFVILHTAFLGNAMRQSRRVPDKGRWIAG 120 An ORF129ng nucleotide sequence <SEQ ED 841 > was predicted to encode a protein having amino acid sequence <SEQ ED 842>: 1 MDFRFDIIYE YRWMFLYGAL TTLGLTVVAT AGGSVLGLLL ALARLIHLEK 51 AGAPMRVLAW ALRKVSLLYV TLFRGTPLFV QIVIWAYVWF PFFVILHTAF 101 LGNAMRQSRR VPDKGRWIAG SLELNCQPRG RKTRGEFPPG ESNLGTEPRN 151 PLSMGQRRFP GCENWYPPQN FIKK* Further work revealed the following gonococcal sequence <SEQ D 843>: 1 ATGGATTTTC gtTTTGACAT TATTTAcgaA TACCGCTGGA TGTTTCTTTA 51 CGGCGCACTG Acgaccttgg ggctgacggt cgtggcgacg gCGGGCGGTT 101 CGGtattggG TCTGTTGTTG GCGTTGGCGC GCCTGATTCA CTTGGAAAAA 151 GCCGGTGCGC CGATGCGCGT GCTGGCGTGG GCGTTGCGTA AGGTTTCGCT 201 GCTGTACGTT ACCCTGTTCC GGGGTACGCC GCTGTTTGTG CAGATTGTGA 251 TTTGGGCGTA TGTGTGGTTT CCGTTTTTCG TCCATCCTTC AGACGGCATT 301 TTGGTCAGCG GCGAGGCGGC AATCGCGCTG CGTCGCGGAT ACGGGCCGCT 351 GATTGCCGGT TCTTTGGCAC TGATCGCCAA CTCGGGGGCG TATATCTGTG 401 AGATTTTCCG CGCGGGCATC CAGTCTATAG ACAAAGGACA GATGGAGGCG 451 GCGTGTTCTT TGGGACTGAC CTATCCGCAG GCGATGCGCT ATGTGATTCT 5 50011 GCCGCAGGCA TTGCGCCGTA TGCTGCCGCC TTTGGCGAGC GAGTTCATCA 551 CGCTCTTGAA AGACAGCTCG CTGCTGTCGG TCATTGCTGT GGCGGAGTTG 601 GCGTATGTTC AGAATACGAT TACGGGCCGG TATTCGGTTT ATGAAGAACC 651 GCTTTACACC GCCGCCCTGA TTTATCTGTT GATGACGACT TTCTTAGGCT 701 GGATATTCCT GCGTTTGGAA AAACGTTACA ATCCGCAACA CCGCTGA This coπesponds to the amino acid sequence <SEQ ID 844; ORF129ng-l>: 1 MDFRFDIIYE YRWMFLYGAL TTLGLTVVAT AGGSVLGLLL ALARLIHLEK 51 AGAPMRVLAW ALRKVSLLYV TLFRGTPLFV QIVIWAYVWF PFFVHPSDGI 101 LVSGEAAIAL RRGYGPLIAG SLALIANSGA YICEIFRAGI QSIDKGQMEA 151 ARSLGLTYPQ AMRYVILPQA LRRMLPPLAS EFITLLKDSS LLSVIAVAEL 201 AYVQNTITGR YSVYEEPLYT VALIYLLMTT FLGWIFLRLE KRYNPQHR* ORF129ng-l and ORF129-1 show 99.2% identity in 248 aa overlap: orf129-1. pep MDFRFDIIYEYRWMFLYGALTTLGLTWATAGGSVLGLLLALARLIHLEKAGAPMRVLAW I II I I I I I I I I I I II I I I II I II I II I I I II I II I I I I I I I I II I I I I II I I I I II II II orfl29ng-l MDFRFDIIYEYRWMFLYGALTTLGLTWATAGGSVLGLLLALARLIHLEKAGAPMRVLAW orf129-1. pep ALRKVSLLYVTLFRGTPLFVQIVIWAYVWFPFFVHPSDGILVSGEAAIALRRGYGPLIAG I II I I I I I I I I I I II I I I II I II I II I II II I II I II II I I II I I I I II I I I II I I I I I I orfl29ng-l ALRKVSLLYVTLFRGTPLFVQIVIWAYVWFPFFVHPSDGILVSGEAAIALRRGYGPLIAG orf129-1.pep SLALIANSGAYICEIFRAGIQSIDKGQMEAARSLGLTYPQAMRYVILPQALRRMLPPLAS I II I I I I I I I I II II I II II I II I II I II II I II II I I I I II I II I I I I I II II I I I I I orfl29ng-l SLALIANSGAYICEIFRAGIQSIDKGQMEAACSLGLTYPQAMRYVILPQALRRMLPPLAS orf129-1. pep EFITLLKDSSLLSVIAVAELAYVQNTITGRYSVYEEPLYTVALIYLLMTTFLGWIFLRLE I I II I I I I I II II I I I I I I II I I II II I II I I I I II II II : I I I I II I I II II I II II II orfl29ng-l EFITLLKDSSLLSVIAVAELAYVQNTITGRYSVYEEPLYTAALIYLLMTTFLGWIFLRLE orf129-1. pep KRYNPQHRX I II I I I I I I orfl29ng-l KRYNPQHRX In addition, ORF129ng-l is homologous to an ABC transporter from A.fulgidus: 2650409 (AE001090) glutamine ABC transporter, permease protein (glnP) [Archaeoglobus fulgidus] ength = 224 Score = 132 bits (329) , Expect = 2e-30 Identities = 86/178 (48%), Positives = 103/178 (57%), Gaps = 18/178 (10%) Query: 65 VSLLYVTLFRGTPLFVQIVIWAYVWFPFFVHPSDGILVSGEAAIALRRGYGPLIAGSLAL 124 +S YV + RGTPL VQI+I +F P+ GI + E A G +AL Sbjct: 58 ISTAYVEVIRGTPLLVQILI VYFGLPAIGINLQPEPA GIIAL 99 Query: 125 IANSGAYICEIFRAGIQSIDKGQMEAACSLGLTYPQAMRYVILPQALRRMLPPLASEFIT 184 SGAYI El RAGI+SI GQMEAA SLG+TY QAMRYVI PQA R +LP L +EFI Sbjct: 100 SICSGAYIAEIVRAGIESIPIGQMEAARSLGMTYLQAMRYVIFPQAFRNILPALGNEFIA 159 Query: 185 LLKDSSLLSVIAVAELAYVQNTITGRYSVYEEPLYTAALIYLLMTTFLGWIFLRLEKR 242 LLKDSSLLSVI++ EL V I P AL YL+MT L + +K+ Sbjct: 160 LLKDSSLLSVISIVELTRVGRQIVNTTFNAWTPFLGVALFYLMMTIPLSRLVAYSQKK 217 This -analysis, including the identification of transmembrane domains in the two proteins, suggests that the proteins from Nmeningitidis and N.gonorrhoeae, and their epitopes, could be useful antigens for vaccines or diagnostics, or for raising antibodies. Example 100 The following partial DNA sequence was identified in N. meningitidis <SEQ ID 845>: 1 .. CTGAAAGAAT GCCGTCTGAA AGACCCTGTT TTTATTCCAA ATATCGTTTA 51 TAAGAACATC GCCATTACTT TCCTGCTCTT GCACGCCGCC GCCGAACTTT 101 GGCTGCCCGC GCAAACCGCC GGTTTTACCG CGCTCGCCGT CGGCTTCATC 151 CTGCTCGCCA AGCTGCGTGA gCTTCACCAT CACGAACTCT TACGTAAACA 201 cTACGTCCGC ACTTATTACy TGCTCCAACT CTTTGCCGCC GCAGgcTAgT 251 TTGTGGACAG GCGCGGCGwA ATTACAAAAC CTGCCCGCyT CCGCGCCCCT 301 GCACCTGATT ACCCTCGGCG GCATGATGGG CGGCGTGATG ATGGTGTGGc 351 TGACCGCCGG ACTGTGGCAC AGCGGCTTTA CCAAACTCGA CTACCCCAAA 401 CTCTGCCGCA TTGCCGTCCC CATCCTTTTC GCCGCCGCCG TCTCGCGCGC 451 TTTCTTGrTG AACGTGAACC CGrTATTTTT CATTACCGTT CCTGCGATTC 501 TGACCGCCGC CGTATTCGTA CTGTATCTTT TCrCGTTTAT ACCGATATTT 551 CGGGCGAATG CGTTTACAGA CGATCCGGAr TAr This coπesponds to the amino acid sequence <SEQ ID 846; ORF130>: 1 .. KECRLKDPV FIPNIVYKNI AITFLLLHAA AELWLPAQTA GFTALAVGFI 51 LLAKLRELHH HELLRKHYVR TYYLLQLFAA AGSLWTGAAX LQNLPASAPL 101 HLITLGGMMG GVMMVWLTAG LWHSGFTKLD YPKLCRIAVP ILFAAAVSRA 151 FLXNVNPXFF ITVPAILTAA VFVLYLFXFI PIFRANAFTD DPE* Further work revealed the complete nucleotide sequence <SEQ ID 847>: 1 ATGCGGCCGT TTTTCGTCGG CGCGGCGGTG CTTGCCATAC TCGGTGCGCT 51 GGTGTTTTTC ATCAACCCCG GTGCCATCGT CCTGCACCGC CAAATTTTCT 101 TGGAACTTAT GCTGCCGGCG GCATACGGCG GTTTTTTGAC TGCGGCTTTG 151 TTGGACTGGA CGGGTTTTTC GGGTAACCTG AAACCTGTCG CGACTTTGAT 201 GGCGGCATTA TTGCTCGCCG CATCCGCTAT ACTGCCCTTT TCGCCGCAAA 251 CTGCCTCGTT TTTCGTCGCC GCCTATTGGC TGGTGTTGCT GCTGTTCTGC 301 GCCCGGCTGA TTTGGCTAGA CCGAAACACC GACAACTTCG CCCTGCTAAT 351 GTTACTTGCC GCGTTCACTG TTTTTCAGAC GGCATATGCC GTCAGCGGCG 401 ATTTGAACCT GTTGCGCGCG CAAGTGCATC TAAATATGGC GGCGGTGATG 451 TTCGTATCCG TGCGCGTCAG TATTCTTTTG GGCGCGGAAG CCCTGAAAGA 501 ATGCCGTCTG AAAGACCCTG TTTTTATTCC AAATATCGTT TATAAAAACA 551 TCGCCATTAC TTTCCTGCTC TTGCACGCCG CCGCCGAACT TTGGCTGCCC 601 GCGCAAACCG CCGGTTTTAC CGCGCTCGCC GTCGGCTTCA TCCTGCTCGC 651 CAAGCTGCGT GAGCTTCACC ATCACGAACT CTTACGTAAA CACTACGTCC 701 GCACTTATTA CCTGCTCCAA CTCTTTGCCG CCGCAGGCTA TTTGTGGACA 751 GGCGCGGCGA AATTACAAAA CCTGCCCGCC TCCGCGCCCC TGCACCTGAT 801 TACCCTCGGC GGCATGATGG GCGGCGTGAT GATGGTGTGG CTGACCGCCG 851 GACTGTGGCA CAGCGGCTTT ACCAAACTCG ACTACCCCAA ACTCTGCCGC 901 ATTGCCGTCC CCATCCTTTT CGCCGCCGCC GTCTCGCGCG CTTTCTTGAT 951 GAACGTGAAC CCGATATTTT TCATTACCGT TCCTGCGATT CTGACCGCCG 1001 CCGTATTCGT ACTGTATCTT TTCACGTTTA TACCGATATT TCGGGCGAAT 1051 GCGTTTACAG ACGATCCGGA ATAA This coπesponds to the amino acid sequence <SEQ ED 848; ORF130-1>: 1 MRPFFVGAAV LAILGALVFF INPGAIVLHR QIFLELMLPA AYGGFLTAAL 51 LDWTGFSGNL KPVATLMAAL LLAASAILPF SPQTASFFVA AYWLVLLLFC 101 ARLIWLDRNT DNFALLMLLA AFTVFQTAYA VSGDLNLLRA QVHLNMAAVM 151 FVSVRVSILL GAEALKECRL KDPVFIPNIV YKNIAITFLL LHAAAELWLP 201 AQTAGFTALA VGFILLAKLR ELHHHELLRK HYVRTYYLLQ LFAAAGYLWT 251 GAAKLQNLPA SAPLHLITLG GMMGGVMMVW LTAGLWHSGF TKLDYPKLCR 301 IAVPILFAAA VSRAFLMNVN PIFFITVPAI LTAAVFVLYL FTFIPIFRAN 351 AFTDDPE* Computer analysis of this amino acid sequence gave the following results: Homology with a predicted ORF from N.meninsitidis (strain Al ORF130 shows 94.3% identity over a 193aa overlap with an ORF (ORF130a) from strain A of N. meningitidis: 10 20 30 orf130. pep LKECRLKDPVFIPNIVYKNIAITFLLLHAA II I I I I I I I I I II I : I I I I I I II I I I I I I I orf130a LNLLRAQVHLNMAAVMFVSVRVSILLGAEALKECRLKDPVFIPNWYKNIAITFLLLHAA 140 150 160 170 180 190 40 50 60 70 80 90 orf130. pep AELWLPAQTAGFTALAVGFILLAKLRELHHHELLRKHYVRTYYLLQLFAAAGSLWTGAAX II I II I I I II II I : I I I I I I I I I II II I I II II I I II I I I I I I II I I I I I II II I I I I orf130a AELWLPAQTAGFTSLAVGFILLAKLRELHHHELLRKHYVRTYYLLQLFAAAGYLWTGAAK 200 210 220 230 240 250 100 110 120 130 140 150 orf130. pep LQNLPASAPLHLITLGGMMGGVMMVWLTAGLWHSGFTKLDYPKLCRIAVPILFAAAVSRA I I II I II II II II I I II I I I: I I I I II I II II I I I I I I I I I I I I II I I I I II II I I I I II orf130a LQNLPASAPLHLITLGGMMGSVMMVWLTAGLWHSGFTKLDYPKLCRIAVPILFAAAVSRA 260 270 280 290 300 310 160 170 180 190 orf130.pep FLXNVNPXFFITVPAILTAAVFVLYLFXFIPIFRANAFTDDPEX I I I II I II I I I II I I I I II I II M: M I I I II I I II I II I I orf130a VLMNVNPIFFITVPAILTAAVFVLYLLTFVPIFRANAFTDDPEX 320 330 340 350 The complete length ORF130a nucleotide sequence <SEQ ID 849> is: 1 ATGCGGCCGT TTTTCGTCGG CGCGGCGGTG CTTGCCATAC TCGGTGCGCT 51 GGTGTTTTTC ATCAACCCCG GTGCCATCGT CCTGCACCGC CAAATTTTCT 101 TGGAACTTAT GCTGCCGGCG GCATACGGCG GTTTTTTGAC TGCGGCTTTG 151 TTGGACTGGA CGGGTTTTTC GGGTAACCTG AAACCTGTCG CGACTTTGAT 201 GGCGGCATTA TTGCTCGCCG CATCCGCTAT ACTGCCCTTT TCGCCGCAAA 251 CTGCCTCGTT TTTCGTCGCC GCCTATTGGC TGGTGTTGCT GCTGTTCTGC 301 GCCCGGCTGA TTTGGCTAGA CCGAAACACC GACAACTTCG CCCTGCTAAT 351 GTTACTTGCC GCGTTCACTG TTTTTCAGAC GGCATATGCC GTCAGCGGCG 401 ATTTGAACCT GTTGCGCGCG CAAGTGCATC TAAATATGGC GGCGGTGATG 451 TTCGTATCCG TGCGCGTCAG TATTCTTTTG GGCGCGGAAG CCCTGAAAGA 501 ATGCCGTCTG AAAGACCCAG TATTCATCCC CAATGTCGTC TATAAAAACA 551 TCGCCATTAC CTTCCTGCTC CTGCACGCCG CCGCCGAACT TTGGCTGCCT 601 GCGCAAACCG CCGGTTTTAC CTCGCTCGCC GTCGGCTTTA TCCTGCTTGC 651 CAAGCTGCGT GAGCTTCACC ATCACGAACT CCTGCGCAAA CACTACGTCC 701 GCACTTATTA CCTGCTCCAA CTCTTTGCCG CCGCAGGCTA TTTGTGGACA 751 GGCGCGGCGA AATTACAAAA CCTGCCCGCC TCCGCGCCCC TGCACCTGAT 801 TACCCTCGGT GGCATGATGG GCAGCGTGAT GATGGTGTGG CTGACTGCCG 851 GACTGTGGCA CAGCGGCTTT ACCAAGCTCG ACTACCCGAA ACTCTGCCGC 901 ATCGCCGTCC CCATCCTNTT CGCCGCCGCC GTTTCGCGCG CTGTTTTAAT 951 GAACGTAAAC CCGATATTCT TCATCACCGT CCCCGCAATT CTGACCGCCG 001 CCGTGTTCGT GCTTTACCTG CTGACATTCG TACCGATCTT TCGGGCGAAC .051 GCGTTTACAG ACGATCCGGA ATAA This encodes a protein having amino acid sequence <SEQ ID 850>: 1 MRPFFVGAAV LAILGALVFF INPGAIVLHR QIFLELMLPA AYGGFLTAAL 51 LDWTGFSGNL KPVATLMAAL LLAASAILPF SPQTASFFVA AYWLVLLLFC 101 ARLIWLDRNT DNFALLMLLA AFTVFQTAYA VSGDLNLLRA QVHLNMAAVM 151 FVSVRVSILL GAEALKECRL KDPVFIPNW YKNIAITFLL LHAAAELWLP 201 AQTAGFTSLA VGFILLAKLR ELHHHELLRK HYVRTYYLLQ LFAAAGYLWT 251 GAAKLQNLPA SAPLHLITLG GMMGSVMMVW LTAGLWHSGF TKLDYPKLCR 301 IAVPILFAAA VSRAVLMNVN PIFFITVPAI LTAAVFVLYL LTFVPIFRAN 351 AFTDDPE* ORF130a and ORF130-1 show 98.3% identity in 357 aa overlap: orf130a. pep MRPFFVGAAVLAILGALVFFINPGAIVLHRQIFLELMLPAAYGGFLTAALLDWTGFSGNL II II I II I I I I I I I II II I I I I II II I I I I I I II II I II I I I II I I I I II I I I II II II I orf130-1 MRPFFVGAAVLAILGALVFFINPGAIVLHRQIFLELMLPAAYGGFLTAALLDWTGFSGNL orf130a . pep KPVATLMAALLLAASAILPFSPQTASFFVAAYWLVLLLFCARLIWLDRNTDNFALLMLLA I II II I I I I I I I I I I I II I I I I II I I I I I I II I II I II I II I I I I I I II I I I I I I I I II I orf130-1 KPVATLMAALLLAASAILPFSPQTASFFVAAYWLVLLLFCARLIWLDRNTDNFALLMLLA orf130a . pep AFTVFQTAYAVSGDLNLLRAQVHLNMAAVMFVSVRVSILLGAEALKECRLKDPVFIPNW I II II I II I I I I II I I I I I I I I II II II I I I I I I I I I I I I I I I I I I I I I I I I I I I I II: I orfl30-l AFTVFQTAYAVSGDLNLLRAQVHLNMAAVMFVSVRVSILLGAEALKECRLKDPVFIPNIV orf130a. pep YKNIAITFLLLHAAAELWLPAQTAGFTSLAVGFILLAKLRELHHHELLRKHYVRTYYLLQ I M II I II I II I II II I I I I I I II II I : I I I I I I I I I I M I I I I II I I I II I I I I I I I I I orf130-1 YKNIAITFLLLHAAAELWLPAQTAGFTALAVGFILLAKLRELHHHELLRKHYVRTYYLLQ orf130a. pep LFAAAGYLWTGAAKLQNLPASAPLHLITLGGMMGSVMMVWLTAGLWHSGFTKLDYPKLCR I I I II I II I I I I II I I I II I I I II I I I I II II II : I I II I II I II I I I II II I II II I I I orf130-1 LFAAAGYLWTGAAKLQNLPASAPLHLITLGGMMGGVMMVWLTAGLWHSGFTKLDYPKLCR orf130a . pep IAVPILFAAAVSRAVLMNVNPIFFITVPAILTAAVFVLYLLTFVPIFRANAFTDDPΞ II I I I II I I I I II I I II I I I I II I I I I II II I I I I I II I : I I : I I I II I I II I II I orf130-1 IAVPILFAAAVSRAFLMNVNPIFFITVPAILTAAVFVLYLFTFIPIFRANAFTDDPE Homology with a predicted ORF from N.εonorrhoeae ORF130 shows 91.7% identity over a 193 aa overlap with a predicted ORF (ORF130ng) from N.gonorrhoeae: orf130. pep LKECRLKDPVFIPNIVYKNIAITFLLLHAA 30 I II II I II I II I II :: I I I I I I I I I I I I I orfl30ng LNLLRAQVHLNMAAVMFVSVRVSVLLGTETLKECRLKDPVFIPNVIYKNIAIT-LLLHAA 201 orf130.pep AELWLPAQTAGFTALAVGFILLAKLRELHHHELLRKHYVRTYYLLQLFAAAGSLWTGAAX 90 I I I I II II I I II I II I II I II II I II I I I I I I II II II I I I I II I I I II I I I II I II I orfl30ng AELWLPAQTAGFTALAVGFILLAKLRELHHHELLRKHYVRTYYLLQLFAAAGYLWTGAAK 261 orf130.pep LQNLPASAPLHLITLGGMMGGVMMVWLTAGLWHSGFTKLDYPKLCRIAVPILFAAAVSRA 150 I I I I I I I II I I I I I II II I I I II I II I II II II I I I I I I II I II II II I I I I: II II I orfl30ng LQNLPASAPLHLITLGGMTGGVMMVWLTAGLWHSGFTKLDYPKLCRIAVSILFASAVSRA 321 orf130. pep FLXNVNPXFFITVPAILTAAVFVLYLFXFIPIFRANAFTDDPE 193 I I I I I I I I I I I I I I I I I I : I I M : I : I I I I I I I I I I I I I orfl30ng VLMNVNPIFFITVPEILTAAVFMLYLLTFVPIFRANAFTDDPE 364 An ORF130ng nucleotide sequence <SEQ LD 851> was predicted to encode a protein having amino acid sequence <SEQ LD 852>: 1 MNKFFTHPMi PFFVGAAVLA ILGALVFFHQ PRRYHPAPPN FLGTYAAGCI 51 RRFFDYRFVG PDGFFRQPET CRYFDGGWA CCGCFIAVFT ATCRIFRRRL 101 LAGVAAVLRL ADLARRQHRT LRSVDVTAAF TVFQTAYAVS GDLNLLRAQV 151 HLNMAAVMFV SVRVSVLLGT ETLKECRLKD PVFIPNVIYK NIAITLLLHA 201 AAELWLPAQT AGFTALAVGF ILLAKLRELH HHELLRKHYV RTYYLLQLFA 251 AAGYLWTGAA KLQNLPASAP LHLITLGGMT GGVMMVWLTA GLWHSGFTKL 301 DYPKLCRIAV SILFASAVSR AVLMNVNPIF FITVPEILTA AVFMLYLLTF 351 VPIFRANAFT DDPE* Further work revealed the following gonococcal DNA sequence <SEQ LD 853>: 1 ATGCGCCCGT TTTTCGTCGG TGCGGCAGTA CTTGCCATAC TCGGTGCGTT 51 GGTGTTTTTT ATCAACCCCG GCGCTATCAT CCTGCACCGC CAAATTTTCT 101 TGGAACTTAT GCTGCCGGCT GCATACGGCG GTTTTTTGAC TACCGCTTTG 151 TTGGACCGGA CGGGTTTTTC AGGCAACCTG AAACCTGCCG CTACTTTGAT 201 GGCGGTGTTG TTGCTTGTTG CGGCTGTTTT ATTGCCGTTT TTACCGCAAC 251 TTGCCGCATT TTTCGTCGCC GCCTATTGGC TGGTGTTGCT GCTGTTCTGC 301 GCCTGGCTGA TTTGGCTCGA CCGCAACACC GACAACTTCG CTCTGTTGAT 351 GTTACTTGCC GCATTTACCG TTTTTCAGAC GGCCTATGCC GTCAGCGGCG 401 ATTTGAACTT ACTGCGCGCG CAAGTGCATT TGAATATGGC GGCGGTCATG 451 TTCGTATCCG TCCGCGTCAG CGTCCTTTTG GGCACGGAAA CCCTGAAAGA 501 ATGCCGTCTG AAAGACCCCG TATTCATCCC CAACGTTATC TATAAAAACA 551 TCGCCATCAC CCTGCTGCTG CACGCCGCCG CCGAACTTTG GCTGCCCGCG 601 CAAACCGCCG GTTTTACTGC GCTTGCCGTC GGCTTCATCC TGCTCGCCAA 651 GCTGCGCGAA CTGCACCATC ACGAACTCTT ACGCAAACAC TACGTCCGCA 701 CTTATTACCT GCTCCAGCTC TTTGCCGCCG CAGGTTATCT GTGGACAGGC 751 GCGGCGAAAC TGCAAAACCT GCCCGCCTCC GCGCCCCTGC ACCTGATTAC 801 CCTCGGCGGC ATGACGGGTG GCGTGATGAT GGTGTGGCTG ACTGCCGGAC 851 TGTGGCACAG CGGCTTTACC AAACTCGACT ACCCGAAACT CTGCCGCATC 901 GCCGTCTCCA TCCTTTTCGC CTCCGCCGTT TCGCGCGCTG TTTTAATGAA 951 CGTGAATCCG ATATTCTTCA TCACCGTTCC CGAGATTCTG ACCGCCGCCG 1001 TGTTCATGCT TTACCTGCTG ACGTTCGTAC CGATTTTTCG AGCGAACGCG 1051 TTTACAGACG ATCCGGAATA A This coπesponds to the .amino acid sequence <SEQ ID 854; ORF130ng-l>: 1 MRPFFVGAAV LAILGALVFF INPGAIILHR QIFLELMLPA AYGGFLTTAL 51 LDRTGFSGNL KPAATLMAVL LLVAAVLLPF LPQLAAFFVA AYWLVLLLFC 101 AWLIWLDRNT DNFALLMLLA AFTVFQTAYA VSGDLNLLRA QVHLNMAAVM 151 FVSVRVSVLL GTETLKECRL KDPVFIPNVI YKNIAITLLL HAAAELWLPA 201 QTAGFTALAV GFILLAKLRE LHHHELLRKH YVRTYYLLQL FAAAGYLWTG 251 AAKLQNLPAS APLHLITLGG MTGGVMMVWL TAGLWHSGFT KLDYPKLCRI 301 AVSILFASAV SRAVLMNVNP IFFITVPEIL TAAVFMLYLL TFVPIFRANA 351 FTDDPE* ORF130ng-l and ORF130-1 show 92.4% identity in 357 aa overlap: orf130-1.pep MRPFFVGAAVLAILGALVFFINPGAIVLHRQIFLELMLPAAYGGFLTAALLDWTGFSGNL I I I II I I I II II I I I II I II I II I I I : II I I I I II II I I I I I II I I I : I I I I I I I II I I orfl30ng-l MRPFFVGAAVLAILGALVFFINPGAIILHRQIFLELMLPAAYGGFLTTALLDRTGFSGNL orf130-1. pep KPVATLMAALLLAASAILPFSPQTASFFVAAYWLVLLLFCARLIWLDRNTDNFALLMLLA II : I I I I I : I I I : I : : : II I II I : I II II I I I I II I I I I II II I I II II II I II I I I orfl30ng-l KPAATLMAVLLLVAAVLLPFLPQLAAFFVAAYWLVLLLFCAWLIWLDRNTDNFALLMLLA orf130-1. pep AFTVFQTAYAVSGDLNLLRAQVHLNMAAVMFVSVRVSILLGAEALKECRLKDPVFIPNIV I I II II I I II I II I I I II I I I I II I I I I II I I II I II : II I : I : I I I I II II I II II I : : orfl30ng-l AFTVFQTAYAVSGDLNLLRAQVHLNMAAVMFVSVRVSVLLGTETLKECRLKDPVFIPNVI orf130-1. pep YKNIAITFLLLHAAAELWLPAQTAGFTALAVGFILLAKLRELHHHELLRKHYVRTYYLLQ I I I II I I II II I I I I I I I I I I II I I I I I I I I I I I II I II II II I I I I II II I I II I I II orfl30ng-l YKNIAIT-LLLHAAAELWLPAQTAGFTALAVGFILLAKLRELHHHELLRKHYVRTYYLLQ orf130-1. pep LFAAAGYLWTGAAKLQNLPASAPLHLITLGGMMGGVMMVWLTAGLWHSGFTKLDYPKLCR I I I I I II II I II I I I II I II I II I II I I II I I I II II I I I II II II I I I II I I II I I I I orfl30ng-l LFAAAGYLWTGAAKLQNLPASAPLHLITLGGMTGGVMMVWLTAGLWHSGFTKLDYPKLCR orf130-1.pep IAVPILFAAAVSRAFLMNVNPIFFITVPAILTAAVFVLYLFTFIPIFRANAFTDDPEX III I I I I : II I II II II II II I I I II I I II I II : I I I : II : I I I I I I II I I II I I orfl30ng-l IAVSILFASAVSRAVLMNVNPIFFITVPEILTAAVFMLYLLTFVPIFRANAFTDDPEX Based on this analysis, it is predicted that the proteins from N. meningitidis and N.gonorrhoeae, and their epitopes, could be useful antigens for vaccines or diagnostics, or for raising antibodies. Example 101 The following partial DNA sequence was identified in N. meningitidis <SEQ ID 855>: 1 ATGGAAATTC GGGCAATAAA ATATACGGCA ATGGCTGCGT TGCTTGCATT 51 TACGGTTGCA GGCTGCCGGC TGGCGGGGTG GTATGAGTGT TCGTCCCTCA 101 CCGGCTGGTG TAAGCCGAGA AAACCGGCTG CCATCGATTT TTGGGATATT 151 GGCGGCGAGA GTCCGCCGTC TTTAGGGGAC TACGAGATAC CGCTTTCAGA 201 CGGCAATAGT TCCGTCAGGG CAAACGAATA TGAATCCGCA CAACAATCTT 251 ACTTTTACAG GAAAATAGGG AAGTTTGAAG C.TGCGGGCT GGATTGGCGT 301 ACGCGTGACG GCAAACCTTT GATTGAGACG TTCAAACAGG GAGGATTTGA 351 CTGCTTGGAA AAG .. This coπesponds to the amino acid sequence <SEQ ID 856; ORF131>: 1 MEIRAIKYTA MAALLAFTVA GCRLAGWYEC SSLTGWCKPR KPAAIDFWDI 51 GGESPPSLGD YEIPLSDGNS SVRANEYESA QQSYFYRKIG KFEXCGLDWR 101 TRDGKPLIET FKQGGFDCLE K.. Further work revealed the complete nucleotide sequence <SEQ ID 857>: 1 ATGGAAATTC GGGCAATAAA ATATACGGCA ATGGCTGCGT TGCTTGCATT 51 TACGGTTGCA GGCTGCCGGC TGGCGGGGTG GTATGAGTGT TCGTCCCTCA 101 CCGGCTGGTG TAAGCCGAGA AAACCGGCTG CCATCGATTT TTGGGATATT 151 GGCGGCGAGA GTCCGCCGTC TTTAGGGGAC TACGAGATAC CGCTTTCAGA 201 CGGCAATCGT TCCGTCAGGG CAAACGAATA TGAATCCGCA CAACAATCTT 251 ACTTTTACAG GAAAATAGGG AAGTTTGAAG CCTGCGGGCT GGATTGGCGT 301 ACGCGTGACG GCAAACCTTT GATTGAGACG TTCAAACAGG GAGGATTTGA 351 CTGCTTGGAA AAGCAGGGGT TGCGGCGCAA CGGTCTGTCC GAGCGCGTCC 401 GATGGTAA This coπesponds to the amino acid sequence <SEQ ID 858; ORF131-l>: 1 MEIRAIKYTA MAALLAFTVA GCRLAGWYEC SSLTGWCKPR KPAAIDFWDI 51 GGESPPSLGD YEIPLSDGNR SVRANEYESA QQSYFYRKIG KFEACGLDWR 101 TRDGKPLIET FKQGGFDCLE KQGLRRNGLS ERVRW* Computer analysis of this amino acid sequence gave the following results: Homology with a predicted ORF from N.meninsitidis (sfrain Al ORF131 shows 95.0% identity over a 121aa overlap with an ORF (ORF131a) from strain A of N. meningitidis: 10 20 30 40 50 60 orf131. pep MEIRAIKYTAMAALLAFTVAGCRLAGWYECSSLTGWCKPRKPAAIDFWDIGGESPPSLGD I I II I I II II I I I II I I I I I II I II I I I I II IM I I I I I I I I I I II II I I II I I I I I I I orf131a MEIRAIKYTAMAALLAFTVAGCRLAGWYECSSLSGWCKPRKPAAIDFWDIGGESPPSLED 10 20 30 40 50 60 70 80 90 100 110 120 orf131. pep YEIPLSDGNSSVRANEYESAQQSYFYRKIGKFEXCGLDWRTRDGKPLIETFKQGGFDCLE II I I II II I I II I II II I I I II I II I II I I I I I I I I I I I II I I I I II I I II MMM orf131a YEIPLSDGNRSVRANEYESAQQSYFYRKIGKFEACGLDWRTRDGKPLIETFKQEGFDCLK 70 80 90 100 110 120 orf131. pep K l orfl31a KQGLRRNGLSERVRWX 130 The complete length ORF 13 la nucleotide sequence <SEQ ID 859> is: 1 ATGGAAATTC GGGCAATAAA ATATACGGCA ATGGCTGCGT TGCTTGCATT 51 TACGGTTGCA GGCTGCCGGT TGGCAGGTTG GTATGAGTGT TCGTCCCTGT 101 CCGGCTGGTG TAAGCCGAGA AAACCTGCCG CCATCGATTT TTGGGATATT 151 GGCGGCGAGA GTCCTCCGTC TTTAGAGGAC TACGAGATAC CGCTTTCAGA 201 CGGCAATCGT TCCGTCAGGG CAAACGAATA TGAATCCGCA CAACAATCTT 251 ACTTTTACAG GAAAATAGGG AAGTTTGAAG CCTGCGGGTT GGATTGGCGT 301 ACGCGTGACG GCAAACCTTT GATTGAGACG TTCAAACAGG AAGGTTTTGA 351 TTGTTTGAAA AAGCAGGGGT TGCGGCGCAA CGGTCTGTCC GAGCGCGTCC 401 GATGGTAA This encodes a protein having amino acid sequence <SEQ ID 860>: 1 MEIRAIKYTA MAALLAFTVA GCRLAGWYEC SSLSGWCKPR KPAAIDFWDI 51 GGESPPSLED YEIPLSDGNR SVRANEYESA QQSYFYRKIG KFEACGLDWR 101 TRDGKPLIET FKQEGFDCLK KQGLRRNGLS ERVRW* ORF131a and ORF131-1 show 97.0% identity in 135 aa overlap: orf131a . pep MEIRAIKYTAMAALLAFTVAGCRLAGWYECSSLSGWCKPRKPAAIDFWDIGGESPPSLED I II I II I II II I I II I II II I I II I I I II II I I : II II I I I I I I I II I I II II I I I I I I orf131-1 MEIRAIKYTAMAALLAFTVAGCRLAGWYECSSLTGWCKPRKPAAIDFWDIGGESPPSLGD orf131a. pep YEIPLSDGNRSVRANEYESAQQSYFYRKIGKFEACGLDWRTRDGKPLIETFKQEGFDCLK I I I I I I I I I I I II I II I I I I I I I I I I II I I I II I I I II I I II I I I I I II I I I I Mill: orf131-1 YEIPLSDGNRSVRANEYESAQQSYFYRKIGKFEACGLDWRTRDGKPLIETFKQGGFDCLE orf131a. pep KQGLRRNGLSERVRWX II I I II I II II I I II I orfl31-l KQGLRRNGLSERVRWX Homology with a predicted ORF from N.εonorrhoeae ORFl 31 shows 89.3% identity over 121 aa overlap with a predicted ORF (ORFl 3 lng) from N.gonorrhoeae: orf131.pep MEIRAIKYTAMAALLAFTVAGCRLAGWYECSSLTGWCKPRKPAAIDFWDIGGESPPSLGD 60 I II M I I I II I II : II II II I I II I II II I I : I I I I I I I I I II I I I I I II II I II I orfl31ng MEIRVIKYTATAALFAFTVAGCRLAGWYECLSLSGWCKPRKPAAIDFWDIGGESPLSLED 60 orf131. pep YEIPLSDGNSSVRANEYESAQQSYFYRKIGKFEXCGLDWRTRDGKPLIETFKQGGFDCLE 120 I I II I I II I I I I I I I I I I I I: II I II I I I I I I I I I I I I I I I II I I : I III II I I I I orf13lng YEIPLSDGNRSVRANEYESAQKSYFYRKIGKFEACGLDWRTRDGKPLVERFKQEGFDCLE 120 orfl31.pep K 121 I orfl31ng KQGLRRNGLSERVRW 134 A complete length ORF131ng nucleotide sequence <SEQ ID 861> was predicted to encode a protein having amino acid sequence <SEQ LD 862>: 1 MEIRVIKYTA TAALFAFTVA GCRLAGWYEC LSLSGWCKPR KPAAIDFWDI 51 GGESPLSLED YEIPLSDGNR SVRANEYESA QKSYFYRKIG KFEACGLDWR 101 TRDGKPLVER FKQEGFDCLE KQGLRRNGLS ERVRW* Further work revealed the following gonococcal DNA sequence <SEQ ID 863>: 1 ATGGAAATTC GGGTAATAAA ATATACGGCA ACGGCTGCGT TGTTTGCATT 51 TACGGTTGCA GGCTGCCGGC TGGCGGGGTG GTATGAGTGT TCGTCCTTGT 101 CCGGCTGGTG TAAGCCGAGA AAACCTGCCG CCATCGATTT TTGGGATATT 151 GGCGGCGAGA GtccgctGTC TTTAGAGGAC TACGAGATAC CGCTTTCAGA 201 CGGCAATCGT TCCGTCAGGG CAAACGAATA TGAATCCGCG CAAAAATCTT 251 ACTTTTATAG GAAAATAGGG AAGTTTGAAG CCTGCGGGTT GGATTGGCGT 301 ACGCGTGACG GCAAACCTTT GGTTGAGAGG TTCAAACAGG AAGGTTTCGA 351 CTGTTTGGAA AAGCAGGGGT TGCGGCGCAA CGGCCTGTCC GAGCGCGTCC 401 GATGGTAA This coπesponds to the amino acid sequence <SEQ ID 864; ORF131ng-l>: 1 MEIRVIKYTA TAALFAFTVA GCRLAGWYEC SSLSGWCKPR KPAAIDFWDI 51 GGESPLSLED YEIPLSDGNR SVRANEYESA QKSYFYRKIG KFEACGLDWR 101 TRDGKPLVER FKQEGFDCLE KQGLRRNGLS ERVRW* ORF131ng-l and ORF131-1 show 92.6% identity in 135 aa overlap: orfl31ng-l.pep MEIRVIKYTATAALFAFTVAGCRLAGWYECSSLSGWCKPRKPAAIDFWDIGGESPLSLED I I I I: I I I II II I : I II I I II I I II II I II I I : I II I II I II I I I 11 I II I I II II I orf131-1 MEIRAIKYTAMAALLAFTVAGCRLAGWYECSSLTGWCKPRKPAAIDFWDIGGESPPSLGD orfl31ng-l.pep YEIPLSDGNRSVRANEYESAQKSYFYRKIGKFEACGLDWRTRDGKPLVERFKQEGFDCLE I II II I I II I I I I I I II II I I : II I I I I II I II II I II I II I I I II I : I III I I II I I orf131-1 YEIPLSDGNRSVRANEYESAQQSYFYRKIGKFEACGLDWRTRDGKPLIETFKQGGFDCLE orfl31ng-l.pep KQGLRRNGLSERVRWX I I I I I I I I I I I I I I I I orfl31-l KQGLRRNGLSERVRWX Based on the presence of a predicted prokaryotic membrane lipoprotein lipid attachment site, it is predicted that the proteins from N. meningitidis and N.gonorrhoeae, and their epitopes, could be useful antigens for vaccines or diagnostics, or for raising antibodies. Example 102 The following partial DNA sequence was identified in N. meningitidis <SEQ ID 865> 1 ATGAAACACA TCCATATTAT CGGTATCGGC GGCACGTTTA TGGGCGGGCT 51 TGCCGCCATT GCCAAAGAAG CGGGGTTTGA AGTCAGCGGT TGCGACGCGA 101 AGATGTATCC GCCGATGAGC ACCCAGCTCG AAGCCTTGGG TATAGACGTG 151 TATGAAGGCT TCGATGCCGC TCAGTTGGAC GAATTTAAAG CCGACGTTTA 201 CGTTATCGGC AATGTCGCCA AGCGCGGGAT GGATGTGGTT GAAGCGATTT 251 TGAACCTCGG CCTGCCtTAT ATtTcCGGCC CGCAATGGCT GTCGGAAAAC 301 GTGCTGCACC ATCATTGGGT ACTCGGTGTG GCGGGGACgC ACGGCAAAAC 351 GACCACCGCC TCCATGCTCG CATGGGTCTT GGAATATgCC GGCCTCGCGC 401 CGGGCTTCCT TATtGGCGGC GTACC.GGAA AATttCGGCG TTTCCGCCCG 451 CCTGCCGCAA ACGCCGCGCC AAGACCCGAA CAGCCAATCG CCGTTTTTcG 501 TCATCGAAGC CGACGAATAC GACACCGCCT TTtTCGACAA ACGTTCTAAA 551 TtCGTGCATT ACCGTCCGCG TACCGCCGTG TTGAACAATC TGGAATTCGA 601 CCACGCCGAC ATCTTTGCCG ACTTGGGCGC GATACAGACc CAGTTCCACT 651 ACCTCGTGCG TACCGTGCCG TCTGAAGGCT TAATCGTCTG CAACGGACGG 701 CAGCAAAGCC TGCAAGATAC TTTGGACAAA GGCTGCTGGA CGCCGGTGGA 751 AAAATTCGGC ACGGAACACG GCTGGCA.. This coπesponds to the amino acid sequence <SEQ LD 866; ORF132>: 1 MKHIHIIGIG GTFMGGLAAI AKEAGFEVSG CDAKMYPPMS TQLEALGIDV 51 YEGFDAAQLD EFKADVYVIG NVAKRGMDW EAILNLGLPY ISGPQWLSEN 101 VLHHHWVLGV AGTHGKTTTA SMLAWVLEYA GLAPGFLIGG VXGKFRRFRP 151 PAANAAPRPE QPIAVFRHRS RRIRHRLFRQ TFXIRALPSA YRRVEQSGIR 201 PRRHLCRLGR DTDPVPLPRA YRAVXRLNRL QRTAAKPARY FGQRLLDAGG 251 KIRHGTRLA.. Further work revealed the complete nucleotide sequence <SEQ ID 867>: 1 ATGAAACACA TCCATATTAT CGGTATCGGC GGCACGTTTA TGGGCGGGCT 51 TGCCGCCATT GCCAAAGAAG CGGGGTTTGA AGTCAGCGGT TGCGACGCGA 101 AGATGTATCC GCCGATGAGC ACCCAGCTCG AAGCCTTGGG TATAGACGTG 151 TATGAAGGCT TCGATGCCGC TCAGTTGGAC GAATTTAAAG CCGACGTTTA 201 CGTTATCGGC AATGTCGCCA AGCGCGGGAT GGATGTGGTT GAAGCGATTT 251 TGAACCTCGG CCTGCCTTAT ATTTCCGGCC CGCAATGGCT GTCGGAAAAC 301 GTGCTGCACC ATCATTGGGT ACTCGGTGTG GCGGGGACGC ACGGCAAAAC 351 GACCACCGCC TCCATGCTCG CATGGGTCTT GGAATATGCC GGCCTCGCGC 401 CGGGCTTCCT TATTGGCGGC GTACCGGAAA ATTTCGGCGT TTCCGCCCGC 451 CTGCCGCAAA CGCCGCGCCA AGACCCGAAC AGCCAATCGC CGTTTTTCGT 501 CATCGAAGCC GACGAATACG ACACCGCCTT TTTCGACAAA CGTTCTAAAT 551 TCGTGCATTA CCGTCCGCGT ACCGCCGTGT TGAACAATCT GGAATTCGAC 601 CACGCCGACA TCTTTGCCGA CTTGGGCGCG ATACAGACCC AGTTCCACTA 651 CCTCGTGCGT ACCGTGCCGT CTGAAGGCTT AATCGTCTGC AACGGACGGC 701 AGCAAAGCCT GCAAGATACT TTGGACAAAG GCTGCTGGAC GCCGGTGGAA 751 AAATTCGGCA CGGAACACGG CTGGCAGGCC GGCGAAGCCA ATGCCGACGG 801 CTCGTTCGAC GTGTTGCTCG ACGGCAAAAC CGCCGGACGC GTCAAATGGG 851 ATTTGATGGG CAGGCACAAC CGCATGAACG CGCTCGCCGT CATTGCCGCC 901 GCGCGTCATG TCGGTGTCGA TATTCAGACC GCCTGCGAAG CCTTGGGCGC 951 GTTTAAAAAC GTCAAACGCC GGATGGAAAT CAAAGGCACG GCAAACGGCA 1001 TCACCGTTTA CGACGACTTC GCCCACCACC CGACCGCCAT CGAAACCACG 1051 ATTCAAGGTT TGCGCCAACG CGTCGGCGGC GCGCGCATCC TCGCCGTCCT 1101 CGAACCGCGT TCCAACACGA TGAAGCTGGG CACGATGAAG TCCGCCCTGC 1151 CTGTAAGCCT CAAAGAAGCC GACCAAGTGT TCTGCTACGC CGGCGGCGTG 1201 GACTGGGACG TCGCCGAAGC CCTCGCGCCT TTGGGCGGCA GGCTGAACGT 1251 CGGCAAAGAC TTCGATGCCT TCGTTGCCGA AATCGTGAAA AACGCCGAAG 1301 TAGGCGACCA TATTTTGGTG ATGAGCAACG GCGGTTTCGG CGGAATACAC 1351 GGAAAGCTGC TGGAAGCTTT GAGATAG This coπesponds to the amino acid sequence <SEQ LD 868; ORF132-l>: 1 MKHIHIIGIG GTFMGGLAAI AKEAGFEVSG CDAKMYPPMS TQLEALGIDV 51 YEGFDAAQLD EFKADVYVIG NVAKRGMDW EAILNLGLPY ISGPQWLSEN 101 VLHHHWVLGV AGTHGKTTTA SMLAWVLEYA GLAPGFLIGG VPENFGVSAR 151 LPQTPRQDPN SQSPFFVIEA DEYDTAFFDK RSKFVHYRPR TAVLNNLEFD 201 HADIFADLGA IQTQFHYLVR TVPSEGLIVC NGRQQSLQDT LDKGCWTPVE 251 KFGTEHGWQA GEANADGSFD VLLDGKTAGR VKWDLMGRHN RMNALAVIAA 301 ARHVGVDIQT ACEALGAFKN VKRRMEIKGT ANGITVYDDF AHHPTAIETT 351 IQGLRQRVGG ARILAVLEPR SNTMKLGTMK SALPVSLKEA DQVFCYAGGV 401 DWDVAEALAP LGGRLNVGKD FDAFVAEIVK NAEVGDHILV MSNGGFGGIH 451 GKLLEALR* Computer analysis of this amino acid sequence gave the following results: Homology with the hypothetical o457 protein of E.coli (accession number U140031 ORF132 and o457 show 58% aa identity in 140 aa overlap: Orf132: 4 IHIIGIGGTFMGGLAAIAKEAGFEVSGCDAKMYPPMSTQLEALGIDVYEGFDAAQLDEFK 63 IHI+GI GTFMGGLA +A++ G EV+G DA +YPPMST LE GI++ +G+DA+QL+ + o457: 3 IHILGICGTFMGGLAMLARQLGHEVTGSDANVYPPMSTLLEKQGIELIQGYDASQLEP-Q 61 Orf132: 64 ADVYVIGNVAKRGMDWEAILNLGLPYISGPQWLSENVLHHHWVLGVAGTHGKTTTASML 123 D+ +IGN RG VEA+L +PY+SGPQWL + VL WVL VAGTHGKTTTA M θ457: 62 PDLVIIGNAMTRGNPCVEAVLEKNIPYMSGPQWLHDFVLRDRWVLAVAGTHGKTTTAGMA 121 Orf132: 124 AWVLEYAGLAPGFLIGGVXG 143 W+LE G PGF+IGGV G o457: 122 TWILEQCGYKPGFVIGGVPG 141 Homology with a predicted ORF from N.meninεitidis (strain Al ORF132 shows 74.6% identity over a 189aa overlap with .an ORF (ORF132a) from strain A of N. meningitidis: 10 20 30 40 50 60 orf132. pep MKHIHIIGIGGTFMGGLAAIAKEAGFEVSGCDAKMYPPMSTQLEALGIDVYEGFDAAQLD I I II I II I II I I I I I I: I I I I I I I I II II I I I I I I I I I I I II I II I I I II II I : I I II orf132a MKHIHIIGIGGTFMGGIAAIAKEAGFEXSGCDAKMYPPMSTQLEALGIGVYEGFDTAQLD 10 20 30 40 50 60 70 80 90 100 110 120 orf132. pep EFKADVYVIGNVAKRGMDWEAILNLGLPYISGPQWLSENVLHHHWVLGVAGTHGKTTTA I II I I I I I II I I I I I I I I I I I I I II II II I II II II : I I I I I I I I I II II I II I I I orf132a EFKADVYVIGNVAKRGMDWEAILNRGLPYISGPQWLAENXLHHHWXLGVAXTHGKTTTA 70 80 90 100 110 120 130 140 150 160 orf132.pep SMLAWVLEYAGLAPGFLIGGVXGKFR RFRPPAANAAPRPEQPI AVFR II I I I I I II I II I I II II II : I M l : I :: I : I I orf132a SMLAWVLEYAGLAPGFXIGGVPENFΞVSARL-PQTPRQDPNSQSPFFVIEADEYDTAFFD 130 140 150 160 170 170 180 190 200 210 220 orf132. pep HRSRRIRHRLFRQTFXIRALPSAYRRVEQSGIRPRRHLCRLGRDTDPVPLPRAYRAVXRL : I I : ::: I orf132a KRSKFVHYRPRTAVLNNLEFDHADIFADLGAIQTQFHHLVRTVPSEGLIVCNGRQQSLQD 180 190 200 210 220 230 The complete length ORF132a nucleotide sequence <SEQ ID 869> is: 1 ATGAAACACA TCCACATTAT CGGTATCGGC GGCACGTTTA TGGGTGGGAT 51 TGCCGCCATT GCCAAAGAAG CAGGGTTTGA ANTCAGCGGT TGCGATGCGA 101 AGATGTATCC GCCGATGAGC ACCCAGCTCG AAGCCTTGGG CATAGGCGTG 151 TATGAAGGCT TCGACACCGC GCAGTTGGAC GAATTTAAAG CCGACGTTTA 201 CGTTATCGGC AATGTCGCCA AGCGCGGGAT GGATGTGGTT GAAGCGATTT 251 TGAACCGTGG GCTGCCTTAT ATTTCCGGCC CGCAATGGCT GGCTGAAAAC 301 NTGCTGCACC ATCATTGGNN ACTCGGCGTG GCGGNGACGC ACGGCAAAAC 351 GACCACCGCG TCTATGCTCG CGTGGGTTTT GGAATATGCC GGACTCGCAC 401 CGGGCTTCNT TATCGGCGGC GTACCGGAAA ACTTCAGCGT TTCCGCCCGC 451 CTGCCGCAAA CGCCGCGCCA AGACCCGAAC AGCCAATCGC CGTTTTTCGT 501 CATTGAAGCC GACGAATACG ACACCGCGTT TTTCGACAAA CGCTCCAAAT 551 TCGTGCATTA CCGTCCGCGT ACCGCCGTGT TGAACAATCT GGAATTCGAC 601 CACGCCGACA TCTTCGCCGA TTTGGGCGCG ATACAGACCC AGTTCCACCA 651 CCTCGTGCGT ACCGTGCCGT CTGAAGGCCT CATCGTCTGC AACGGACGGC 701 AGCAAAGCCT GCAAGACACT TTGGACAAAG GCTGCTGGAC GCCGGTGGAA 751 AAATTCGGCA CGGAACACGG CTGGCAGGCC GGCGAAGCCA ATGCCGATGG 801 CTCGTTCGAC GTGTTGCTTG ACGGCAAAAA AGCCGGACAC GTCGCTTGGA 851 GTTTGATGGG CGGACACAAC CGCATGAACG CGCTCGCNGT CATCGCCGCC 901 GCGCGTCATG CCGGAGTNGA CATTCAGACG GCCTGCGAAG CCTTGAGCAC 951 GTTTAAAAAC GTCAAACGCC GCATGGAAAT CAAAGGCACG GCAAACGGTA 1001 TCACCGTTTA CGACGACTTC GCCCACCATC CGACCGCTAT CGAAACCACG 1051 ATTCAAGGTT TGCGCCAGCG CGTCGGCGGC GCGCGCATCC TCGCCGTCCT 1101 CGAACCGCGT TCCAATACGA TGAAGCTGGG TACGATGAAA GCCGCCCTGC 1151 CCGCAAGCCT CAAAGAAGCC GACCAAGTGT TCTGNTACGC CGGCGGCGCG 1201 GACTGGGACG TTGCCGAAGC CCTCGCGCCT TTGGGCGGCA GGCTGCACGT 1251 CGGCAAAGAC TTCGATGCCT TCGTTGCCGA AATCGTGAAA AACGCCGAAG 1301 CAGGCGACCA TATTTTGGTG ATGAGCAACG GCGGTTTCGG CGGAATACAC 1351 ACCAAACTGC TGGACGCTTT GAGATAG This encodes a protein having amino acid sequence <SEQ LD 870>: 1 MKHIHIIGIG GTFMGGIAAI AKEAGFEXSG CDAKMYPPMS TQLEALGIGV 51 YEGFDTAQLD EFKADVYVIG NVAKRGMDW EAILNRGLPY ISGPQWLAEN 101 XLHHHWXLGV AXTHGKTTTA SMLAWVLEYA GLAPGFXIGG VPENFSVSAR 151 LPQTPRQDPN SQSPFFVIEA DEYDTAFFDK RSKFVHYRPR TAVLNNLEFD 201 HADIFADLGA IQTQFHHLVR TVPSEGLIVC NGRQQSLQDT LDKGCWTPVE 251 KFGTEHGWQA GEANADGSFD VLLDGKKAGH VAWSLMGGHN RMNALAVIAA 301 ARHAGVDIQT ACEALSTFKN VKRRMEIKGT ANGITVYDDF AHHPTAIETT 351 IQGLRQRVGG ARILAVLEPR SNTMKLGTMK AALPASLKEA DQVFXYAGGA 401 DWDVAEALAP LGGRLHVGKD FDAFVAEIVK NAEAGDHILV MSNGGFGGIH 451 TKLLDALR* ORF132a and ORF132-1 show 93.9% identity in 458 aa overlap: orf132a. pep MKHIHIIGIGGTFMGGIAAIAKEAGFEXSGCDAKMYPPMSTQLEALGIGVYEGFDTAQLD I II I I I II II I I II II : I I II I I I I II I I I II II I I I I II II I II I I I II I I I: I I I I orf132-1 MKHIHIIGIGGTFMGGLAAIAKEAGFEVSGCDAKMYPPMSTQLEALGIDVYEGFDAAQLD orf132a . pep EFKADVYVIGNVAKRGMDWEAILNRGLPYISGPQWLAENXLHHHWXLGVAXTHGKTTTA II I II II I I I I I II II II II I II I I I II II II I I II : I I I II II I II I I I I I I I I I orfl32-l EFKADVYVIGNVAKRGMDWEAI NLGLPYISGPQWLSENVLHHHWVLGVAGTHGKTTTA orf132a . pep SMLAWVLEYAGLAPGFXIGGVPENFSVSARLPQTPRQDPNSQSPFFVIEADEYDTAFFDK II II II I II II I II II I II I II I I : I I I I I II I I II II I I I I I II I I II II I I I I I I II orf132-1 SMLAWVLEYAGLAPGFLIGGVPENFGVSARLPQTPRQDPNSQSPFFVIEADEYDTAFFDK orf132a . pep RSKFVHYRPRTAVLNNLEFDHADIFADLGAIQTQFHHLVRTVPSEGLIVCNGRQQSLQDT I I II II I II II II I I I I I II I II I II I I I II II I I I: I I II II I I I I I I I II I I II I I II orf132-1 RSKFVHYRPRTAVLNNLEFDHADIFADLGAIQTQFHYLVRTVPSEGLIVCNGRQQSLQDT orf132a . pep LDKGCWTPVEKFGTEHGWQAGEANADGSFDVLLDGKKAGHVAWSLMGGHNRMNALAVIAA I I I II I I I I I I I I I II I II II I II I I II II II I I II MM Mill I II I I I I I I II I orf132-1 LDKGCWTPVEKFGTEHGWQAGEANADGSFDVLLDGKTAGRVKWDLMGRHNRMNALAVIAA orf132a . pep ARHAGVDIQTACEALSTFKNVKRRMEIKGTANGITVYDDFAHHPTAIETTIQGLRQRVGG I I I : I I I I I I I II I I : : I I I I II I I I I II I I II II I II I I I I I I I I I II I I II II I I I I I orf132-1 ARHVGVDIQTACEALGAFKNVKRRMEIKGTANGITVYDDFAHHPTAIETTIQGLRQRVGG orf132a . pep ARILAVLEPRSNTMKLGTMKAALPASLKEADQVFXYAGGADWDVAEALAPLGGRLHVGKD II I II I I I I I I I I I I II II I : I I I : I II I I I I I I II I I : I I II II I I II II I I I : I I II orf132-1 ARILAVLEPRSNTMKLGTMKSALPVSLKEADQVFCYAGGVDWDVAEALAPLGGRLNVGKD orf132a. pep FDAFVAEIVKNAEAGDHILVMSNGGFGGIHTKLLDALRX I II I II I I I II I I : II I I II I I I I I II I I I I I I : I I I I orf 132-1 FDAFVAEIVKNAEVGDHILVMSNGGFGGIHGKLLEALRX Homology with a predicted ORF from Nεonorrhoeae ORF 132 shows 89.6% identity over 259 aa overlap with a predicted ORF (ORF132ng) from N. gonorrhoeae: orf132.pep MKHIHIIGIGGTFMGGLAAIAKEAGFEVSGCDAKMYPPMSTQLEALGIDVYEGFDAAQLD 60 I II II I I I II I II II I : I I I II I I II: I I I II I I I I I II I I I I I I II I I : I II I I I I I: orfl32ng MKHIHIIGIGGTFMGGIAAIAKEAGFKVSGCDAKMYPPMSTQLEALGIGVHEGFDAAQLE 60 orf132.pep EFKADVYVIGNVAKRGMDWEAILNLGLPYISGPQWLSENVLHHHWVLGVAGTHGKTTTA 120 I I: I I: II I II I M II II II II I II I II I II II I I I : I I I II I I I II I I I I I I II I I II orfl32ng EFQADIYVIGNVARRGMDWEAILNRGLPYISGPQWLAENVLHHHWVLGVAGTHGKTTTA 120 orf132. pep SMLAWVLEYAGLAPGFLIGGVXGKFRRFRPPAANAAPRPEQPIAVFRHRSRRIRHRLFRQ 180 II I II I I I I I II I I I II II II II II I I I I I: I I I I II I I II I I I I I II I I II II II I orfl32ng SMLAWVLEYAGLAPGFLIGGVPGKFRRFRPPTANAASRPEQQIAVFRHRSRRIRHRLFRQ 180 orf132. pep TFXIRALPSAYRRVEQSGIRPRRHLCRLGRDTDPVPLPRAYRAVXRLNRLQRTAAKPARY 240 M MM II I II II I I II I II I I I I I I I II II I I I I M :: I : I II I II II II II orfl32ng TLQIRALSPAYRRVEQSGIRPRRHLRRLGRDTDPVPPPRAHRTIRRPHRLQRTAAKPARY 240 orf132. pep FGQRLLDAGGKIRHGTRLA 259 II I II I I I II II I I II I I orfl32ng FGQRLLDAGGKIRHRTRLADW 261 An ORF132ng nucleotide sequence <SEQ ED 871 > was predicted to encode a protein having amino acid sequence <SEQ LD 872>: 1 MKHIHIIGIG GTFMGGIAAI AKEAGFKVSG CDAKMYPPMS TQLEALGIGV 51 HEGFDAAQLE EFQADIYVIG NVARRGMDW EAILNRGLPY ISGPQWLAEN 101 VLHHHWVLGV AGTHGKTTTA SMLAWVLEYA GLAPGFLIGG VPGKFRRFRP 151 PTANAASRPE QQIAVFRHRS RRIRHRLFRQ TLQIRALSPA YRRVEQSGIR 201 PRRHLRRLGR DTDPVPPPRA HRTIRRPHRL QRTAAKPARY FGQRLLDAGG 251 KIRHRTRLAD W* Further work revealed the following gonococcal DNA sequence <SEQ ID 873>: 1 ATGAAACACA TCCACATTAT CGGTATCGGC GGCACGTTTA TGGGCGGGAT 51 TGCCGCCATT GCCAAAGAAG CCGGGTTCAA AGTCAGCGGT TGCGACGCGA 101 AGATGTATCC GCCGATGAGC ACCCAGCTCG AAGCCTTGGG CATAGGCGTA 151 CACGAAGGCT TCGATGCCGC GCAGTTGGAA GAATTTCAAG CCGATATTTA 201 CGTCATCGGC AATGTCGCCA GGCGCGGGAT GGATGTGGTC GAGGCGATTT 251 TGAACCGTGG GCTGCCTTAT ATTTCCGGCC CGCAATGGCT GGCTGAAAac 301 GTGCtgcacc atcaTTGGgt ACTCGGCGTG GcagggaCGC ACGGcaaAac 351 gaccaCcGcg tCCATGCTCG CCTGGGTCTT GGAATATGCC GGACTCGCGC 401 CGGGCTTCCT CATCGGCGGt gtaccggaAA ATTTCGGCGT TTCCGCCCGC 451 CTACCGCAAA CGCCGCGTCA AGACCCGAAC AGCAAATCGC CGTTTTTCGT 501 CATCGAAGCC GACGAATACG ACACCGCCTT TTTCGACAAA CGCTCCAAAT 551 TCGTGCATTA TCGCCCGCGT ACCGCCGTGT TGAACAATCT GGAATTCGAC 601 CACGCCGACA TCTTCGCCGA CTTGGGCGCG ATACAGACCC AGTTCCACCA 651 CCTCGTGCGC ACCGTACCAT CCGAAGGCCT CATCGTCTGC AACGGACAGC 701 AGCAAAGCCT GCAAGATACT TTGGACAAAG GCTGCTGGAC GCCGGTGGAA 751 AAATTCGGCA CCGGACACGG CTGGCAGATT GGTGAAGTCA ATGCCGACGG 801 CTCGTTCGAC GTATTGCTTG ACGGCAAAAA AGCCGGACAC GTCGCATGGG 851 ATTTGATGGG CGGACACAAC CGCATGAACG CGCTCGCCGT CATCGCTGCC 901 GCACGCCATG CCGGAGTCGA TGTTCAGACG GCCTGCGAAG CCTTGGGTGC 951 GTTTAAAAAC GTCAAACGCC GCATGGAAAT CAAAGGCACG GCAAACGGCA 1001 TCACCGTTTA CGACGATTTC GCCCACCACC CGACCGCCAT CGAAACCACG 1051 ATTCAAGGTT TGCGCCAACG TGTCGGCGGC GCGCGCATCC TCGCCGTCCT 1101 CGAGCCGCGT TCCAACACCA TGAAACTCGG CACGATGAAG TCCGCCCTGC 1151 CCGCAAGCCT CAAAGAAGCC GACCAAGTGT TCTGCTACGC CGGCGGCGCG 1201 GACTGGGACG TTGCCGAAGC CCTCGCGCCT TTGGGCTGCA GGCTGCGCGT 1251 CGGTAAAGAT TTCGATACCT TCGTTGCCGA AATTGTGAAA AACGCCCGAA 1301 CCGGCGACCA TATTTTGGTG ATGAGCAACG GCGGTTTCGG CGGAATACAC 1351 ACCAAACTGC TGGACGCTTT GAGATAG This coπesponds to the amino acid sequence <SEQ LD 874; ORF132ng-l>: 1 MKHIHIIGIG GTFMGGIAAI AKEAGFKVSG CDAKMYPPMS TQLEALGIGV 51 HEGFDAAQLE EFQADIYVIG NVARRGMDW EAILNRGLPY ISGPQWLAEN 101 VLHHHWVLGV AGTHGKTTTA SMLAWVLEYA GLAPGFLIGG VPENFGVSAR 151 LPQTPRQDPN SKSPFFVIEA DEYDTAFFDK RSKFVHYRPR TAVLNNLEFD 201 HADIFADLGA IQTQFHHLVR TVPSEGLIVC NGQQQSLQDT LDKGCWTPVE 251 KFGTGHGWQI GEVNADGSFD VLLDGKKAGH VAWDLMGGHN RMNALAVIAA 301 ARHAGVDVQT ACEALGAFKN VKRRMEIKGT ANGITVYDDF AHHPTAIETT 351 IQGLRQRVGG ARILAVLEPR SNTMKLGTMK SALPASLKEA DQVFCYAGGA 401 DWDVAEALAP LGCRLRVGKD FDTFVAEIVK NARTGDHILV MSNGGFGGIH 451 TKLLDALR* ORF132ng-l and ORF132-1 show 93.2% identity in 458 aa overlap: orfl32ng-l.pep MKHIHIIGIGGTFMGGIAAIAKEAGFKVSGCDAKMYPPMSTQLEALGIGVHEGFDAAQLE I II II I I II I I II II I : I I I I II I II : I I I I I II I I II I I I I I II I I I I : I II I I I I I : orf132-1 MKHIHIIGIGGTFMGGLAAIAKEAGFEVSGCDAKMYPPMSTQLEALGIDVYEGFDAAQLD orf 132ng-l . pep EFQADIYVIGNVARRGMDWEAILNRGLPYISGPQWLAENVLHHHWVLGVAGTHGKTTTA I I : I I : I I I I I I I : I I I I I I I I I I I I I I I I I I I I I I : I I I I I I I I I I I I I I I I I I I I I I orf 132-1 EFKADVYVIGNVAKRGMDVVEAILNLGLPYISGPQWLSENVLHHHWVLGVAGTHGKTTTA orfl32ng-l . pep SMLAWVLEYAGLAPGFLIGGVPENFGVSARLPQTPRQDPNSKSPFFVIEADEYDTAFFDK I I II I I II I II I II I I I I I I II I II I II I I I I II I I I I I I I : I I I I I I I I I I II II M I I orf132-1 SMLAWVLEYAGLAPGFLIGGVPENFGVSARLPQTPRQDPNSQSPFFVIEADEYDTAFFDK orfl32ng-l.pep RSKFVHYRPRTAVLNNLEFDHADIFADLGAIQTQFHHLVRTVPSEGLIVCNGQQQSLQDT I II I I II I II I II I I I II I II I II I I I I II II II II : I I I I I I II I I II I II: II I I I I I orf132-1 RSKFVHYRPRTAVLNNLEFDHADIFADLGAIQTQFHYLVRTVPSEGLIVCNGRQQSLQDT orf132ng-l . pep LDKGCWTPVEKFGTGHGWQIGEVNADGSFDVLLDGKKAGHVAWDLMGGHNRMNALAVIAA I II II I I II I I II I I II I I I: I I I I I II II II II MM I I I I I I I II I II I I I I I orfl32-l LDKGCWTPVEKFGTEHGWQAGEANADGSFDVLLDGKTAGRVKWDLMGRHNRMNALAVIAA orf132ng-l . pep ARHAGVDVQTACEALGAFKNVKRRMEIKGTANGITVYDDFAHHPTAIETTIQGLRQRVGG II M II I : I I I I I II I I I II I I I I I II I II II II II II I I I I I II I I I I I II II II II I I orfl32-1 ARHVGVDIQTACEALGAFKNVKRRMEIKGTANGITVYDDFAHHPTAIETTIQGLRQRVGG orf132ng-l . pep ARILAVLEPRSNTMKLGTMKSALPASLKEADQVFCYAGGADWDVAEALAPLGCRLRVGKD I I II II II I I II I II I I I II I I I I : I I I I I II II II II I : I I I I I I I I I I II II II I I orf132-1 ARILAVLEPRSNTMKLGTMKSALPVSLKEADQVFCYAGGVDWDVAEALAPLGGRLNVGKD orfl32ng-l.pep FDTFVAEIVKNARTGDHILVMSNGGFGGIHTKLLDALRX I I : I I I II I I II :: I I I I II I I I I I II I II I II : I I I I orf132-1 FDAFVAEIVKNAEVGDHILVMSNGGFGGIHGKLLEALRX In addition, ORF132ng-l is homologous to a hypothetical E.coli protein: pir||S56459 hypothetical protein o457 - Escherichia coli >gi 1537075 (U14003) 0RF_o457 [Escherichia coli] >gi 11790680 (AE000494) hypothetical 48.5 kD protein in fbp-p ba intergenic region [Escherichia coli] Length = 457 Score = 474 bits (1207), Expect = e-133 Identities = 249/439 (56%), Positives = 294/439 (66%), Gaps = 13/439 (2%) Query: 22 KEAGFKVSGCDAKMYPPMSTQLEALGIGVHEGFDAAQLEEFQADIYVIGNVARRGMDWE 81 ++ G +V+G DA +YPPMST LE GI + +G+DA+QLE Q D+ +IGN RG VE Sbjct: 21 RQLGHEVTGSDANVYPPMSTLLEKQGIELIQGYDASQLEP-QPDLVIIGNAMTRGNPCVE 79 Query: 82 AILNRGLPYISGPQWLAENVLHHHWVLGVAGTHGKTTTASMLAWVLEYAGLAPGFLIGGV 141 A+L + +PY+SGPQWL + VL WVL VAGTHGKTTTA M W+LE G PGF+IGGV Sbjct: 80 AVLEKNIPYMSGPQWLHDFVLRDRWVLAVAGTHGKTTTAGMATWILEQCGYKPGFVIGGV 139 Query: 142 PENFGVSARLPQTPRQDPNSKSPFFVIEADEYDTAFFDKRSKFVHYRPRTAVLNNLEFDH 201 P NF VSA L +S FFVIEADEYD AFFDKRSKFVHY PRT +LNNLEFDH Sbjct: 140 PGNFEVSAHL GESDFFVIEADEYDCAFFDKRSKFVHYCPRTLILNNLEFDH 190 Query: 202 ADIFADLGAIQTQFHHLVRTVPSEGLIVCNGQQQSLQDTLDKGCWTPVEKFGTGHGWQIG 261 ADIF DL AIQ QFHHLVR VP +G 1+ +L+ T+ GCW+ E G WQ Sbjct: 191 ADIFDDLKAIQKQFHHLVRIVPGQGRIIWPENDINLKQTMAMGCWSEQELVGEQGHWQAK 250 Query: 262 EVNADGS-FDVLLDGKKAGHVAWDLMGGHNRMNALAVIAAARHAGVDVQTACEALGAFKN 320 ++ D S ++VLLDG+K G V W L+G HN N L IAAARH GV A ALG+F N Sbjct: 251 KLTTDASEWEVLLDGEKVGEVKWSLVGEHNMHNGLMAIAAARHVGVAPADAANALGSFIN 310 Query: 321 VKRRMEIKGTANGITVYDDFAHHPTAIETTIQGLRQRVGG-ARILAVLEPRSNTMKLGTM 379 +RR+E++G ANG+TVYDDFAHHPTAI T+ LR +VGG ARI+AVLEPRSNTMK+G Sbjct: 311 ARRRLELRGEANGVTVYDDFAHHPTAILATLAALRGKVGGTARIIAVLEPRSNTMKMGIC 370 Query: 380 KSALPASLKEADQVF-CYAGGADWDVAEALAPLGCRLRVGKDFDTFVAEIVKNARTGDHI 438 K L SL AD+VF W VAE D DT +VK A+ GDHI Sbjct: 371 KDDLAPSLGRADEVFLLQPAHIPWQVAEVAEACVQPAHWSGDVDTLADMWKTAQPGDHI 430 Query: 439 LVMSNGGFGGIHTKLLDAL 457 LVMSNGGFGGIH KLLD L Sbjct: 431 LVMSNGGFGGIHQKLLDGL 449 Based on this analysis, it was predicted that these proteins from N. meningitidis .and N.gonorrhoeae, and their epitopes, could be useful antigens for vaccines or diagnostics, or for raising antibodies. ORF 132-1 (26.4kDa) was cloned in pET and pGex vectors and expressed in E.coli, as described above. The products of protein expression and purification were analyzed by SDS-PAGE. Figure 20A shows the results of affinity purification of the His-fusion protein, and Figure 20B shows the results of expression of the GST-fusion in E.coli. Purified His-fusion protein was used to immunise mice, whose sera were used for FACS analysis (Figure 20C) and ELISA (positive result). These experiments confirm that ORF132 is a surface-exposed protein, and that it is a useful immunogen. Example 103 The following partial DNA sequence was identified in N. meningitidis <SEQ LD 875> 1 .. CCGGGCTATT ACGGCTCGGA TGACGAATTT AAGCGGGCAT TCGGAGAAAA 51 CTCGCCGACA TmCAAGAAAC ATTGCAACCG GAGCTGCGGG ATTTATGAAC 101 CCGTATTGAA AAAATACGGC AAAAAGCGCG CCAACAACCA TTCGGTCAGC 151 ATTAGTGCGG ACTTCGGCGA TTATTTCATG CCGTTCGCCA GCTATTCGCG 201 CACACACCGT ATGCCCAACA TCCAAGAAAT GTATTTTTCC CAAATCGGCG 251 ACTCCGGCGT TCACACCGCC TTAAAACCAG AGCGCGCAAA CACTTGGCAA 301 TTTGGCTTCr ATACCTATAA AAAAGGATTG TTAAAACAAG ATGATACATT 351 AGGATTAAAA CTGGTCGGCT ACCGCAGCCG CATCGACAAC TACATCCACA 401 ACGTTTACGG GAAATGGTGG GATTTGAACG GGGATATTCC GAGCTGGGTC 451 AGCAGCACCG GGCTTGCCTA CACCATCCAA CATCGCrATT TCAwAGACAA 501 AGTGCATCAA nnnnnnnnnn nnnnnnnnnn nnnnTACGAT TATGGGCGTT 551 TTTTCACCAA CCTTTCTTAC GCCTATCAAA AAAGCACGCA ACCGACCAAC 601 TTCAGCGATG CGAGCGAATC GCCCAACAAT GCGTCCAAAG AAGACCAACT 651 CAAACAAGGT TATGGGTTGA GCAGGGTTTC CGCCCTGCCG CGAGATTACG 701 GACGTTTGGA AGTCGGTACG CGCTGGTTGG GCAACAAACT GACTTTGGGC 751 GGCGCGATGC GCTATTTCGG CAAGAGCATC CGCGCGACGG CTGAAGAACG 801 CTATATCGAC GGCACCAACG GGGGAAATAC CAGCAATTTC CGGCAACTGG 851 GCAAGCGTTC CATCAAACAA ACCGAAACTC TTGCCCGCCA GCCTTTGATT 901 TTwGATTTTa ACGCCGCTTA CGAGCCGAAG AAAAACCTTA TTTTCCGCGC 951 CGAAGTCAAA AATCTGTTCG ACAGGCGTTA TATCGATCCG CTCGATGCGG 1001 GCAATGATGC GGCAAC.GAG CGTTATTACA GCTCGTTCGA CCCGAAAGAC 1051 AAGGACrrAG ACGTAACGTG TAATGCTGAT AAAACGTTGT GCaACGGCAA 1101 ATACGGCGGC ACAAGCAAAA GCGTATTGAC CAATTTTGCA CGCGGACGCA 1151 CCTTTTTgAT GACGATGAGC TACAAGTTTT AA This coπesponds to the amino acid sequence <SEQ ID 876; ORF133>: 1 . . PGΪΥGSDDEF KRAFGENSPT XKKHCNRSCG IYEPVLKKYG KKRANNHSVS 51 ISADFGDYFM PFASYSRTHR MPNIQEMYFS QIGDSGVHTA LKPERANTWQ 101 FGFXTYKKGL LKQDDTLGLK LVGYRSRIDN YIHNVYGKWW DLNGDIPSWV 151 SSTGLAYTIQ HRXFXDKVHQ XXXXXXXXYD YGRFFTNLSY AYQKSTQPTN 201 FSDASESPNN ASKEDQLKQG YGLSRVSALP RDYGRLEVGT RWLGNKLTLG 251 GAMRYFGKSI RATAEERYID GTNGGNTSNF RQLGKRSIKQ TETLARQPLI 301 XDFNAAYEPK KNLIFRAEVK NLFDRRYIDP LDAGNDAAXE RYYSSFDPKD 351 KDXDVTCNAD KTLCNGKYGG TSKSVLTNFA RGRTFLMTMS YKF* Further work revealed the further partial DNA sequence <SEQ ID 877>: 1 GAGGCGCAGA TACAGGTTTT GGAAGATGTG CACGTCAAGG CGAAGCGCGT 51 ACCGAAAGAC AAAAAAGTGT TTACCGATGC GCGTGCCGTA TCGACCCGTC 101 AGGATATATT CAAATCCAGC GAAAACCTCG ACAACATCGT ACGCAGCATC 151 CCCGGTGCGT TTACACAGCA AGATAAAAGC TCGGGCATTG TGTCTTTGAA 201 TATTCGCGGC GACAGCGGGT TCGGGCGGGT CAATACGATG GTGGACGGCA 251 TCACGCAGAC CTTTTATTCG ACTTCTACCG ATGCGGGCAG GGCAGGCGGT 301 TCATCTCAAT TCGGTGCATC TGTCGACAGC AATTTTATTG CCGGACTGGA 351 TGTCGTCAAA GGCAGCTTCA GCGGCTCGGC AGGCATCAAC AGCCTTGCCG 401 GTTCGGCGAA TCTGCGGACT TTAGGCGTGG ATGACGTCGT TCAGGGCAAT 451 AATACCTACG GCCTGCTGCT AAAAGGTCTG ACCGGCACCA ATTCAACCAA 501 AGGTAATGCG ATGGCGGCGA TAGGTGCGCG CAAATGGCTG GAAAGCGGAG 551 CATCTGTCGG TGTGCTTTAC GGGCACAGCA GGCGCAGCGT GGCGCAAAAT 601 TACCGCGTGG GCGGCGGCGG GCAGCACATC GGAAATTTTG GCGCGGAATA 651 TTTGGAACGG CGCAAGCAGC GATATTTTGT ACAAGAGGGT GCTTTGAAAT 701 TCAATTCCGA CAGCGGAAAA TGGGAGCGGG ATTTACAAAG GCAACAGTGG 751 AAATACAAGC CGTATAAAAA TTACAACAAC CAAGAACTAC AaAAATACAT 801 CGAAGAGCAT GACAAAAGCT GGCGGGAAAA CCTg . CaCCG CAATACGACA 851 TTACCCCCAT CGATCCGTCC AGCCTGAAGC AGCAGTCGGC AGGCAATCTG 901 TTTAAATTGG AATACGACGG CGTATTCAAT AAATACACGG CGCAATTTCG 951 CGATTTAAAC ACCAAAATCG GCAGCCGCAA AATCATCAAC CGCAATTATC 1001 AGTTCAATTA CGGTTTGTCT TTGAACCCGT ATACCAACCT CAATCTGACC 1051 GCAGCCTACA ATTCGGGCAG GCAGAAATAT CCGAAAGGGT CGAAGTTTAC 1101 AGGCTGGGGG CTTTTAAAGG ATTTTGAAAC CTACAACAAC GCGAAAATCC 1151 TCGACCTCAA CAACACCGCC ACCTTCCGGC TGCCCCGCGA AACCGAGTTG 1201 CAAACCACTT TGGGCTTCAA TTATTTCCAC AACGAATACG GCAAAAACCG 1251 CTTTCCTGAA GAATTGGGGC TGTTTTTCGA CGGTCCTGAT CAGGACAACG 1301 GGCTTTATTC CTATTTGGGG CGGTTTAAGG GCGATAAAGG GCTGCTGCCC 1351 CAAAAATCAA CCATTGTCCA ACCGGCCGGC AGCCAATATT TCAACACGTT 1401 CTACTTCGAT GCCGCGCTCA AAAAAGACAT TTACCGCTTA AACTACAGCA 1451 CCAATACCGT CGGCTACCGT TTCGGCGGCG AATATACGGG CTATTACGGC 1501 TCGGATGACG AATTTAAGCG GGCATTCGGA GAAAACTCGC CGACATACAA 1551 GAAACATTGC AACCGGAGCT GCGGGATTTA TGAACCCGTA TTGAAAAAAT 1601 ACGGCAAAAA GCGCGCCAAC AACCATTCGG TCAGCATTAG TGCGGACTTC 1651 GGCGATTATT TCATGCCGTT CGCCAGCTAT TCGCGCACAC ACCGTATGCC 1701 CAACATCCAA GAAATGTATT TTTCCCAAAT CGGCGACTCC GGCGTTCACA 1751 CCGCCTTAAA ACCAGAGCGC GCAAACACTT GGCAATTTGG CTTCAATACC 1801 TATAAAAAAG GATTGTTAAA ACAAGATGAT ACATTAGGAT TAAAACTGGT 1851 CGGCTACCGC AGCCGCATCG ACAACTACAT CCACAACGTT TACGGGAAAT 1901 GGTGGGATTT GAACGGGGAT ATTCCGAGCT GGGTCAGCAG CACCGGGCTT 1951 GCCTACACCA TCCAACATCG CAATTTCAAA GACAAAGTGC ACAAACACGG 2001 TTTTGAGTTG GAGCTGAATT ACGATTATGG GCGTTTTTTC ACCAACCTTT 2051 CTTACGCCTA TCAAAAAAGC ACGCAACCGA CCAACTTCAG CGATGCGAGC 2101 GAATCGCCCA ACAATGCGTC CAAAGAAGAC CAACTCAAAC AAGGTTATGG 2151 GTTGAGCAGG GTTTCCGCCC TGCCGCGAGA TTACGGACGT TTGGAAGTCG 2201 GTACGCGCTG GTTGGGCAAC AAACTGACTT TGGGCGGCGC GATGCGCTAT 2251 TTCGGCAAGA GCATCCGCGC GACGGCTGAA GAACGCTATA TCGACGGCAC 2301 CAACGGGGGA AATACCAGCA ATTTCCGGCA ACTGGGCAAG CGTTCCATCA 2351 AACAAACCGA AACTCTTGCC CGCCAGCCTT TGATTTTTGA TTTTTACGCC 2401 GCTTACGAGC CGAAGAAAAA CCTTATTTTC CGCGCCGAAG TCAAAAATCT 2451 GTTCGACAGG CGTTATATCG ATCCGCTCGA TGCGGGCAAT GATGCGGCAA 2501 CGCAGCGTTA TTACAGCTCG TTCGACCCGA AAGACAAGGA CGAAGACGTA 2551 ACGTGTAATG CTGATAAAAC GTTGTGCAAC GGCAAATACG GCGGCACAAG 2601 CAAAAGCGTA TTGACCAATT TTGCACGCGG ACGCACCTTT TTGATGACGA 2651 TGAGCTACAA GTTTTAA This coπesponds to the amino acid sequence <SEQ ID 878; ORF133-1 > • 1 EAQIQVLEDV HVKAKRVPKD KKVFTDARAV STRQDIFKSS ENLDNIVRSI 51 PGAFTQQDKS SGIVSLNIRG DSGFGRVNTM VDGITQTFYS TSTDAGRAGG 101 SSQFGASVDS NFIAGLDWK GSFSGSAGIN SLAGSANLRT LGVDDWQGN 151 NTYGLLLKGL TGTNSTKGNA MAAIGARKWL ESGASVGVLY GHSRRSVAQN 201 YRVGGGGQHI GNFGAEYLER RKQRYFVQEG ALKFNSDSGK WERDLQRQQW 251 KYKPYKNYNN QELQKYIEEH DKSWRENLXP QYDITPIDPS SLKQQSAGNL 301 FKLEYDGVFN KYTAQFRDLN TKIGSRKIIN RNYQFNYGLS LNPYTNLNLT 351 AAYNSGRQKY PKGSKFTGWG LLKDFETYNN AKILDLNNTA TFRLPRETEL 401 QTTLGFNYFH NEYGKNRFPE ELGLFFDGPD QDNGLYSYLG RFKGDKGLLP 451 QKSTIVQPAG SQYFNTFYFD AALKKDIYRL NYSTNTVGYR FGGEYTGYYG 501 SDDEFKRAFG ENSPTYKKHC NRSCGIYEPV LKKYGKKRAN NHSVSISADF 551 GDYFMPFASY SRTHRMPNIQ EMYFSQIGDS GVHTALKPER ANTWQFGFNT 601 YKKGLLKQDD TLGLKLVGYR SRIDNYIHNV YGKWWDLNGD IPSWVSSTGL 651 AYTIQHRNFK DKVHKHGFEL ELNYDYGRFF TNLSYAYQKS TQPTNFSDAS 701 ESPNNASKED QLKQGYGLSR VSALPRDYGR LEVGTRWLGN KLTLGGAMRY 751 FGKSIRATAE ERYIDGTNGG NTSNFRQLGK RSIKQTETLA RQPLIFDFYA 801 AYEPKKNLIF RAEVKNLFDR RYIDPLDAGN DAATQRYYSS FDPKDKDEDV 851 TCNADKTLCN GKYGGTSKSV LTNFARGRTF LMTMSYKF* Computer analysis of this amino acid sequence gave the following results: Homology with with the probable TonB-dependent receptor HL121 of H.influenzae (accession number U32801) ORF133 and HI121 show 57% aa identity in 363aa overlap: Orf133: 31 IYEPVLKKYGKKRANNHSVSISADFGDYFMPFASYSRTHRMPNIQEMYFSQIGDSGVHTA 90 I EP+L K G K+A NHS ++SA+ DYFMPF +YSRTHRMPNIQEM+FSQ+ ++GV+TA HI121: 563 INEPILHKSGHKKAFNHSATLSAELSDYFMPFFTYSRTHRMPNIQEMFFSQVSNAGVNTA 622 Orf133: 91 LKPERANTWQFGFXTYKKGLLKQDDTLGLKLVGYRSRIDNYIHNVYGKWWDLNGDIPSWV 150 LKPE+++T+Q GF TYKKGL QDD LG+KLVGYRS I NYIHNVYG WW +P+W HI121: 623 LKPEQSDTYQLGFNTYKKGLFTQDDVLGVKLVGYRSFIKNYIHNVYGVWW—RDGMPTWA 680 Orf133: 151 SSTGLAYTIQHRXFXDKVHXXXXXXXXXYDYGRFFTNLSYAYQKSTQPTNFSDASESPNN 210 S G YTI H+ + V YD GRFF N+SYAYQ++ QPTN++DAS PNN HI121: 681 ESNGFKYTIAHQNYKPIVKKSGVELEINYDMGRFFANVSYAYQRTNQPTNYADASPRPNN 740 Orf133: 211 ASKEDQLKQGYGLSRVSALPRDYGRLEVGTRWLGNKLTLGGAMRYFGKSIRATAEERYID 270 AS+ED LKQGYGLSRVS LP+DYGRLE+GTRW KLTLG A RY+GKS RAT EE YI+ HI121: 741 ASQEDILKQGYGLSRVSMLPKDYGRLELGTRWFDQKLTLGLAARYYGKSKRATIEEEYIN 800 Orf133: 271 GTNGGNTSNFRQLGKRSIKQTETLARQPLIXDFNAAYEPKKNLIFRAEVKNLFDRRYIDP 330 G+ + R+ ++K+TE + +QP+I D + +YEP K+LI +AEV+NL D+RY+DP HI121: 801 GSR-FKKNTLRRENYYAVKKTEDIKKQPIILDLHVSYEPIKDLIIKAEVQNLLDKRYVDP 859 Orf133: 331 LDAGNDAAXERYYSSFDPKDKDXDVTCNADKTLCNGKYGGTSKSVLTNFARGRTFLMTMS 390 LDAGNDAA +RYYSS + + C D + C GG+ K+VL NFARGRT++++++ HI121: 860 LDAGNDAASQRYYSSL NNSIECAQDSSAC GGSDKTVLYNFARGRTYILSLN 910 Orf133: 391 YKF 393 YKF HI121: 911 YKF 913 Homology with a predicted ORF from N.meninsitidis (sfrain Al ORF133 shows 90.8% identity over a 392aa overlap with an ORF (ORF133a) from strain A of N. meningitidis: 10 20 30 orf133.pep PGYYGSDDEFKRAFGENSPTXKKHCNRSCGI III I I II I I II I I I I I I I II I I: II II orf133a FYFDAALKKDIYRLNYSTNTVGYRFGGXYTGYYXSDDEFKRAFGENSPTYXKHCNQSCGI 450 460 470 480 490 500 40 50 60 70 80 90 orf133. pep YEPVLKKYGKKRANNHSVSISADFGDYFMPFASYSRTHRMPNIQEMYFSQIGDSGVHTAL I I II II I II II I II I I II I I I I I I II I II II I II I I II II I I I I I II I I I I II II II II I orf133a YEPVLKKYGKKRANNHSVSISADFGDYFMPFASYSRTHRMPNIQEMYFSQIGDSGVHTAL 510 520 530 540 550 560 100 110 120 130 140 150 orf133. pep KPERANTWQFGFXTYKKGLLKQDDTLGLKLVGYRSRIDNYIHNVYGKWWDLNGDIPSWVS II I I I I I II I I I I I I I I I II II I I II I I I I I I II I I I I II I I I II I I I I I : I I I I I I orf133a KPERANTWQFGFNTYKKGLLKQDDILGLKLVGYRSRIDXYIHNVYGKWWDLNGNIPSWVS 570 580 590 600 610 620 160 170 180 190 200 210 orf133. pep STGLAYTIQHRXFXDKVHQXXXXXXXXYDYGRFFTNLSYAYQKSTQPTNFSDASESPNNA I II I I I I I I I I I I I II: III I I I I I I I I I I I II II I I I I II I I I I I I II orf133a STGLAYTIQHRNFKDKVHKHGFELELNYDYXRFFTNLSYAYQKSTQPTNFSDASESPNNA 630 640 650 660 670 680 220 230 240 250 260 270 orf133. pep SKEDQLKQGYGLSRVSALPRDYGRLEVGTRWLGNKLTLGGAMRYFGKSIRATAEERYIDG I I I I I I I I I I I II I II II I I I I I I II I I II I I I II I I II I I I I I I I II I I I I II I I I I I orf133a SKEDQLKQGYGLSRVSALPRDYGRLEVGTRWLGNKLTLGGAMRYFGKSIRATAEERYIDX 690 700 710 720 730 740 280 290 300 310 320 330 orf133.pep TNGGNTSNFRQLGKRSIKQTETLARQPLIXDFNAAYEPKKNLIFRAEVKNLFDRRYIDPL III II II I I I I I I I I I I I I I II II I I I I I I I I I I I II I II I II I I II I I I I I I orf133a TNGXXTSNFRQLGKRSIXQTETLARQPLIFDXYAAYEPKKXLIFRAEVKNLFDRRYIDPL 750 760 770 780 790 800 340 350 360 370 380 390 orf133. pep DAGNDAAXERYYSSFDPKDKDXDVTCNADKTLCNGKYGGTSKSVLTNFARGRTFLMTMSY I I I I II I :: II I II II I I I I I Mill I : I I I I I II I I I I II I I I I I I I I I MM I I I orf133a DAGNDAATQRYYSSFDPKDKDEEVTCNDDNTLCNGKYGGTSKSVLTNFARGXTFLITMSY 810 820 830 840 850 860 orf133. pep KFX I I I orfl33a KFX 870 A partial ORF 133a nucleotide sequence <SEQ ID 879> is: 1 AAAGACAAAA AAGTGTTTAC CGATGCGCGT GCCGTATCGA CCCGTCAGGA 51 TATATTCAAA TCCANCGAAA ACCTCGACAA CATCGTACGC ANCATCCCCG 101 GTGCGTTTAC ACANCAANAT AAAAGCTCGG GCNTTGTGTC TTTGAATATT 151 CGCNGCGACA GCGGGTTCGG GCGGGTCAAT ACNATGGTNG ACGGCATCAC 201 NCANACCTTT TATTCGACTT CTACCGATGC GGGCAGGGCA GGCGGTTCAT 251 CTCAATTCGG TGCATCTGTC GACAGCAATT TTATNGCCGG ACTGGATGTC 301 GTCAAAGGCA GCTTCAGCGG CTCGGCAGGC ATCAACAGCC TTGCCGGTTC 351 GGCGAATCTG CGGACTTTAN GCGTGGATGA TGTCGTTCAG GGCAATANTA 401 CNTACGGCCT GCTGCTAAAA GGTCTGACCG GCACCAATTC AACCAAAGGT 451 AATGCGATGG CGGCGATAGG TGCGCGCAAA TGGCTGGAAA GCGGAGCATC 501 TGTCGGTGTG CTTTACGGGC ACAGCAGGCG CAGCGTGGCG CAAAATTACC 551 GCGTGGGCGG CGGCGGGCAG CACATCGGAA ATTTTGGCGC GGAATATCTG 601 GAACGACGCA AGCAACGATA TTTTGAGCAA GAAGGCGGGT TGAAATTCAA 651 TTCCAACAGC GGAAAATGGG AGCGGGATTT CCAAAAGTCG TACTGGAAAA 701 CCAAGTGGTA TCAAAAATAC GATGCCCCCC AAGAACTGCA AAAATACATC 751 GAAGGTCATG ATAAAAGCTG GCGGGAAAAC CTGGCGCCGC AATACGACAT 801 CACCCCCATC GATCCGTCCA GCCTGAAGCN GCAGTCGGCA GGCAACCTGT 851 TTAAATTGGA ATACGACGGC GTATTCAATA AATACACGGC GCAATTTCGC 901 GATTTAAACA CCAAAATCGG CAGCCGCAAA ATCATCAACC GCAATTATCA 951 ATTCAATTAC GGTTTGTCTT TGAACCCGTA TACCAACCTC AATCTGACCG 1001 CAGCCTACAA TTCGGGCAGG CAGAAATATC CGAAAGGGTC GAAGTTTACA 1051 GGCTGGGGGC TTTTNAAAGA TTTTGAAACC TACAACAACG CAAAAATCCT 1101 CGACCTCANC AACACCTCCA CCTTCCGGCT GCCCCGTGAA ACCGAGTTGC 1151 AAACCACTTT GGGCTTCAAT TATTTCCACA ACGAATACGG CAAAAACCGC 1201 TTTCCTGAAG AATTGGGGCT GTTTTTCGAC GGTCCGGATC ANGACAACGG 1251 GCTTTATTCC TATTTGGGGC GGTTTAAGGG CGATAAAGGG CTGCTGCCCC 1301 AAAAATCAAC CATTGTCCAA CCGGCCGGCA GCCAATATTT CAACACGTTC 1351 TACTTCGATG CCGCGCTCAA AAAAGACATT TACCGCTTAA ACTACAGCAC 1401 CAATACCGTC GGCTACCGTT TCGGCGGCNA ATATACGGGC TATTACNGCT 1451 CGGATGACGA ATTTAAGCGG GCATTCGGAG AAAACTCGCC GACATACANG 1501 AAACATTGCA ACCAGAGCTG CGGAATTTAT GAACCCGTAT TGAAAAAATA 1551 CGGCAAAAAG CGCGCCAACA ACCATTCGGT CAGCATTAGT GCGGACTTCG 1601 GCGATTATTT CATGCCGTTC GCCAGCTATT CGCGCACACA CCGTATGCCC 1651 AACATCCAAG AAATGTATTT TTCCCAAATC GGCGACTCCG GCGTTCACAC 1701 CGCCTTAAAA CCAGAGCGCG CAAACACTTG GCAATTTGGC TTCAATACCT 1751 ATAAAAAAGG ATTGTTAAAA CAAGATGATA TATTAGGATT AAAACTGGTC 1801 GGCTACCGCA GCCGCATCGA CNACTACATC CACAACGTTT ACGGGAAATG 1851 GTGGGATTTG AACGGGAATA TTCCGAGCTG GGTCAGCAGC ACCGGGCTTG 1901 CCTACACCAT CCAACACCGC AATTTCAAAG ACAAAGTGCA CAAACACGGT 1951 TTTGAGTTGG AGCTGAATTA CGATTATNGG CGTTTTTTCA CCAACCTTTC 2001 TTACGCCTAT CAAAAAAGCA CGCAACCGAC CAACTTCAGC GATGCGAGCG 2051 AATCGCCCAA CAATGCGTCC AAAGAAGACC AACTCAAACA AGGTTATGGG 2101 TTGAGCAGGG TTTCCGCCCT GCCGCGAGAT TACGGACGTT TGGAAGTCGG 2151 TACGCGCTGG TTGGGCAACA AACTGACTTT GGGCGGCGCG ATGCGCTATT 2201 TCGGCAAGAG CATCCGCGCG ACGGCTGAAG AACGCTATAT CGACGNCACC 2251 AATGGGGNAN NTACCAGCAA TTTCCGGCAA CTGGGCAAGC GTTCCATCAN 2301 ACAAACCGAA ACCCTTGCCC GCCAGCCTTT GATTTTTGAT TTNTACGCCG 2351 CTTACGAGCC GAAGAAAAAN CTTATTTTCC GCGCCGAAGT CAAAAATCTG 2401 TTCGACAGGC GTTATATCGA TCCGCTCGAT GCGGGCAATG ATGCGGCAAC 2451 GCAGCGTTAT TACAGTTCGT TCGACCCGAA AGACAAGGAC GAAGAAGTAA 2501 CGTGTAATGA TGATAACACG TTATGCAACG GCAAATACGG CGGCACAAGC 2551 AAAAGCGTAT TGACCAATTT TGCACGCGGA CNCACCTTTT TGATAACGAT 2601 GAGCTACAAG TTTTAA This encodes a protein having (partial) amino acid sequence <SEQ ID 880>: 1 KDKKVFTDAR AVSTRQDIFK SXENLDNIVR XIPGAFTXQX KSSGXVSLNI 51 RXDSGFGRVN TMVDGITXTF YSTSTDAGRA GGSSQFGASV DSNFXAGLDV 101 VKGSFSGSAG INSLAGSANL RTLXVDDWQ GNXTYGLLLK GLTGTNSTKG 151 NAMAAIGARK WLESGASVGV LYGHSRRSVA QNYRVGGGGQ HIGNFGAEYL 201 ERRKQRYFEQ EGGLKFNSNS GKWERDFQKS YWKTKWYQKY DAPQELQKYI 251 EGHDKSWREN LAPQYDITPI DPSSLKXQSA GNLFKLEYDG VFNKYTAQFR 301 DLNTKIGSRK IINRNYQFNY GLSLNPYTNL NLTAAYNSGR QKYPKGSKFT 351 GWGLXKDFET YNNAKILDLX NTSTFRLPRE TELQTTLGFN YFHNEYGKNR 401 FPEELGLFFD GPDXDNGLYS YLGRFKGDKG LLPQKSTIVQ PAGSQYFNTF 451 YFDAALKKDI YRLNYSTNTV GYRFGGXYTG YYXSDDEFKR AFGENSPTYX 501 KHCNQSCGIY EPVLKKYGKK RANNHSVSIS ADFGDYFMPF ASYSRTHRMP 551 NIQEMYFSQI GDSGVHTALK PERANTWQFG FNTYKKGLLK QDDILGLKLV 601 GYRSRIDXYI HNVYGKWWDL NGNIPSWVSS TGLAYTIQHR NFKDKVHKHG 651 FELELNYDYX RFFTNLSYAY QKSTQPTNFS DASESPNNAS KEDQLKQGYG 701 LSRVSALPRD YGRLEVGTRW LGNKLTLGGA MRYFGKSIRA TAEERYIDXT 751 NGXXTSNFRQ LGKRSIXQTE TLARQPLIFD XYAAYEPKKX LIFRAEVKNL 801 FDRRYIDPLD AGNDAATQRY YSSFDPKDKD EEVTCNDDNT LCNGKYGGTS 851 KSVLTNFARG XTFLITMSYK F* ORF133a and ORF133-1 show 94.3% identity in 871 aa overlap: 10 20 30 40 orf133a . pep KDKKVFTDARAVSTRQDIFKSXENLDNIVRXIPGAFTXQXKS II I I II I I I I II II I II II I I I I I I I II I I II I I I I II orf133-1 EAQIQVLEDVHVKAKRVPKDKKVFTDARAVSTRQDIFKSSENLDNIVRSIPGAFTQQDKS 10 20 30 40 50 60 50 60 70 80 90 100 orf133a . pep SGXVSLNIRXDSGFGRVNTMVDGITXTFYSTSTDAGRAGGSSQFGASVDSNFXAGLDWK II I I I II I II I I I II I II I I I I I I I I I I II I I II II I II I I II I I I II I I I I I I II orf133-1 SGIVSLNIRGDSGFGRVNTMVDGITQTFYSTSTDAGRAGGSSQFGASVDSNFIAGLDWK 70 80 90 100 110 120 110 120 130 140 150 160 orf133a. pep GSFSGSAGINSLAGSANLRTLXVDDWQGNXTYGLLLKGLTGTNSTKGNAMAAIGARKWL I II I I I II II I I I I I I I I II I I I I I I I I I II I I I I II I I I II I I I I I II II II II I I I orfl33-1 GSFSGSAGINSLAGSANLRTLGVDDWQGNNTYGLLLKGLTGTNSTKGNAMAAIGARKWL 130 140 150 160 170 180 170 180 190 200 210 220 orf133a . pep ESGASVGVLYGHSRRSVAQNYRVGGGGQHIGNFGAEYLERRKQRYFEQEGGLKFNSNSGK II I I I II II I I II I II I II I I II I I I II II II I I I I I II I II II II I I I : II I I I : I II orf133-1 ESGASVGVLYGHSRRSVAQNYRVGGGGQHIGNFGAEYLERRKQRYFVQEGALKFNSDSGK 190 200 210 220 230 240 230 240 250 260 270 280 orf133a . pep WERDFQKSYWKTKWYQKYDAPQELQKYIEGHDKSWRENLAPQYDITPIDPSSLKXQSAGN MIMM: M I MM: I I II II I I II I II II II II I I I I I I I II II I I I I II orf133-1 WERDLQRQQWKYKPYKNYNN-QELQKYIEEHDKSWRENLXPQYDITPIDPSSLKQQSAGN 250 260 270 280 290 290 300 310 320 330 340 orf133a . pep LFKLEYDGVFNKYTAQFRDLNTKIGSRKIINRNYQFNYGLSLNPYTNLNLTAAYNSGRQK II II I II II I I II I II I I I I I II I I I I I I II I I I I I I I I I I II I I II I I I II I I I I I II I orf133-1 LFKLEYDGVFNKYTAQFRDLNTKIGSRKIINRNYQFNYGLΞLNPYTNLNLTAAYNSGRQK 300 310 320 330 340 350 350 360 370 380 390 400 orf133a . pep YPKGSKFTGWGLXKDFETYNNAKILDLXNTSTFRLPRETELQTTLGFNYFHNEYGKNRFP II I II II I I I I I I I I I I I I I I I I I II I I : I I II II II I I I II II I I II I II I I I I I II orf133-1 YPKGSKFTGWGLLKDFETYNNAKILDLNNTATFRLPRETELQTTLGFNYFHNEYGKNRFP 360 370 380 390 400 410 410 420 430 440 450 460 orf133a . pep EELGLFFDGPDXDNGLYSYLGRFKGDKGLLPQKSTIVQPAGSQYFNTFYFDAALKKDIYR II I II I I I I I I I I I I I I II II II I II I I I II I I II I I I I I I II II I II I I I II II I I I I orf133-1 EELGLFFDGPDQDNGLYSYLGRFKGDKGLLPQKSTIVQPAGSQYFNTFYFDAALKKDIYR 420 430 440 450 460 470 470 480 490 500 510 520 orf133a . pep LNYSTNTVGYRFGGXYTGYYXSDDEFKRAFGENSPTYXKHCNQSCGIYEPVLKKYGKKRA I I I I I I I I II I I II I I II I I II I I II I I I I I II II I I II : I I I I I II I I I II II I I I orf133-1 LNYSTNTVGYRFGGEYTGYYGSDDEFKRAFGENSPTYKKHCNRSCGIYEPVLKKYGKKRA 480 490 500 510 520 530 530 540 550 560 570 580 orf133a . pep NNHSVSISADFGDYFMPFASYSRTHRMPNIQEMYFSQIGDSGVHTALKPERANTWQFGFN I I II II I I I I I I I II I II II I II I II I I I II I I I I II I I I I I I I I I I I I I II I II I II I I orf133-1 NNHSVSISADFGDYFMPFASYSRTHRMPNIQEMYFSQIGDSGVHTALKPERANTWQFGFN 540 550 560 570 580 590 590 600 610 620 630 640 orf133a . pep TYKKGLLKQDDILGLKLVGYRSRIDXYIHNVYGKWWDLNGNIPSWVSSTGLAYTIQHRNF II II I II II II II I II I I II I II I I I I I I I I I II I I II : II I I I I I I II I II II I I I I orf133-1 TYKKGLLKQDDTLGLKLVGYRSRIDNYIHNVYGKWWDLNGDIPSWVSSTGLAYTIQHRNF 600 610 620 630 640 650 650 660 670 680 690 700 orf133 . pep KDKVHKHGFELELNYDYXRFFTNLSYAYQKSTQPTNFSDASESPNNASKEDQLKQGYGLS I II I I I I I I I II I I I I I I I I I I II II I I I I I I II I I I I II I I II I II I II I II I I I I II orf133-1 KDKVHKHGFELELNYDYGRFFTNLSYAYQKSTQPTNFSDASESPNNASKEDQLKQGYGLS 660 670 680 690 700 710 710 720 730 740 750 760 orf133a. pep RVSALPRDYGRLEVGTRWLGNKLTLGGAMRYFGKSIRATAEERYIDXTNGXXTSNFRQLG I I II I I II II I II I II II I II I I I I I II II II I I I II I I I II II II III I I I I I I II orf133-1 RVSALPRDYGRLEVGTRWLGNKLTLGGAMRYFGKSIRATAEERYIDGTNGGNTSNFRQLG 720 730 740 750 760 770 770 780 790 800 810 820 orf133a . pep KRSIXQTETLARQPLIFDXYAAYEPKKXLIFRAEVKNLFDRRYIDPLDAGNDAATQRYYS orf133-1 KRSIKQTETLARQPLIFDFYAAYEPKKNLIFRAEVKNLFDRRYIDPLDAGNDAATQRYYS 780 790 800 810 820 830 830 840 850 860 870 orf133a . pep SFDPKDKDEEVTCNDDNTLCNGKYGGTSKSVLTNFARGXTFLITMSYKFX II II II I II : I I I I I : I II I I I I I I II I I II I II I II I I I : II II I II orfl33-l SFDPKDKDEDVTCNADKTLCNGKYGGTSKSVLTNFARGRTFLMTMSYKFX 840 850 860 870 880 Homology with a predicted ORF from N.gonorrhoeae ORF133 shows 92.3% identity over 392 aa overlap with a predicted ORF (ORF133ng) from N. gonorrhoeae: orf133. pep PGYYGSDDEFKRAFGENSPTXKKHCNRSCGI 31 I I I II :: I I I I I II I I I I : IMM MM orfl33ng FYFDAALKKDIYRLNYSTNAINYRFGGEYTGYYGSENEFKRAFGENSPAYKEHCDPSCGL 560 orf133. pep YEPVLKKYGKKRANNHSVSISADFGDYFMPFASYSRTHRMPNIQEMYFSQIGDSGVHTAL 91 I I II I II I I II I II I I I II I I II I II I II I I I : I I I I II I I II I I I I I II I I I I I II I II orfl33ng YEPVLKKYGKKRANNHSVSISADFGDYFMPFAGYSRTHRMPNIQEMYFSQIGDSGVHTAL 620 orf133.pep KPERANTWQFGFXTYKKGLLKQDDTLGLKLVGYRSRIDNYIHNVYGKWWDLNGDIPSWVS 151 II I I I I I I I I II II II I I II I I I II I II II I I I II I I I II I II I I I I I I I I II I I I I: orfl33ng KPERANTWQFGFNTYKKGLLKQDDILGLKLVGYRSRIDNYIHNVYGKWWDLNGDIPSWVG 680 orf133.pep STGLAYTIQHRXFXDKVHQXXXXXXXXYDYGRFFTNLSYAYQKSTQPTNFSDASESPNNA 211 II I I I II I : I I I IM I I II II II I I I I I I I II II II II I II I II I I I I orfl33ng STGLAYTIRHRNFKDKVHKHGFELELNYDYGRFFTNLSYAYQKSTQPTNFSDASESPNNA 740 orf133.pep SKEDQLKQGYGLSRVSALPRDYGRLEVGTRWLGNKLTLGGAMRYFGKSIRATAEERYIDG 271 II II I II II I II II I II I I I I I I II I I I I II I I I I I I II II I I I I I I I II I I I I I II I I I orfl33ng SKEDQLKQGYGLSRVSALPRDYGRLEVGTRWLGNKLTLGGAMRYFGKSIRATAEERYIDG 800 orf133.pep TNGGNTSNFRQLGKRSIKQTETLARQPLIXDFNAAYEPKKNLIFRAEVKNLFDRRYIDPL 331 I I II I I II II I II I I I I I I I I I II I I I I II I I I I II I I I I I II I I II I I II II II II orfl33ng TNGGNTSNVRQLGKRSIKQTETLARQPLIFDFYAAYEPKKNLIFRAEVKNLFDRRYIDPL 860 orf133.pep DAGNDAAXERYYSSFDPKDKDXDVTCNADKTLCNGKYGGTSKSVLTNFARGRTFLMTMSY 391 II I II IM: II I I I II I I I I I I II I I I I II II II I I I I II I II I II I I I I I I I I II I I I orfl33ng DAGNDAATQRYYSSFDPKDKDEDVTCNADKTLCNGKYGGTSKSVLTNFARGRTFLMTMSY 920 orfl33.pep KF 393 I I orfl33ng KF 922 The complete length ORF133ng nucleotide sequence <SEQ ID 881> is predicted to encode a protein having amino acid sequence <SEQ ID 882>: 1 MRSSFRLKPI CFYLMGVMLY HHSYAEDAGR AGSEAQIQVL EDVHVKAKRV 51 PKDKKVFTDA RAVSTRQDVF KSGENLDNIV RSIPGAFTQQ DKSSGIVSLN 101 IRGDSGFGRV NTMVDGITQT FYSTSTDAGR AGGSSQFGAS VDSNFIAGLD 151 WKGSFSGSA GINSLAGSAN LRTLGVDDW QGNNTYGLLL KGLTGTNSTK 201 GNAMAAIGAR KWLESGASVG VLYGHSRRGV AQNYRVGGGG QHIGNFGEEY 251 LERRKQQYFV QEGGLKFNAG SGKWERDLQR QYWKTKWYKK YEDPQELQKY 301 IEEHDKSWRE NLAPQYDITP IDPSGLKQQS AGNLINLEYD GVFNKYTAQF 351 RDLNTRIGSR KIINRNYQFN YGLSLNPYTN LNLTAAYNSG RQKYPKGAKF 401 TGWGLLKDFE TYNNAKILDL NNTATFRLPR ETELQTTLGF NYFHNEYGKN 451 RFPEELGLFF DGPDQDNGLY SYLGRFKGDK GLLPQKΞTIV QPAGSQYFNT 501 FYFDAALKKD IYRLNYSTNA INYRFGGEYT GYYGSENEFK RAFGENSPAY 551 KEHCDPSCGL YEPVLKKYGK KRANNHSVSI SADFGDYFMP FAGYSRTHRM 601 PNIQEMYFSQ IGDSGVHTAL KPERANTWQF GFNTYKKGLL KQDDILGLKL 651 VGYRSRIDNY IHNVYGKWWD LNGDIPSWVG STGLAYTIRH RNFKDKVHKH 701 GFELELNYDY GRFFTNLSYA YQKSTQPTNF SDASESPNNA SKEDQLKQGY 751 GLSRVSALPR DYGRLEVGTR WLGNKLTLGG AMRYFGKSIR ATAEERYIDG 801 TNGGNTSNVR QLGKRSIKQT ETLARQPLIF DFYAAYEPKK NLIFRAEVKN 851 LFDRRYIDPL DAGNDAATQR YYSSFDPKDK DEDVTCNADK TLCNGKYGGT 901 SKSVLTNFAR GRTFLMTMSY KF* A variant was also identified, being encoded by the gonococcal DNA sequence <SEQ LD 883>: 1 ATGAGATCTT CTTTCCGGTT GAAGCCGATT TGTTTTTATC TTATGGGTGT 51 TATGCTATAT CATCATAGTT ATGCCGAAGA TGCAGGGCGC GCGGGCAGCG 101 AGGCGCAGAT ACAGGTTTTG GAAGATGTGC ACGTCAAGGC GAAGCGCGTA 151 CCGAAAGACA AAAAAGTGTT TACCGATGCG CGTGCCGTAT CGACCCGTca 201 gGATGTGTTC AAATCCGGCG AAAACCTCGA CAACATCGTA CGCAGCATAC 251 CCGGTGCGTT TACACAGCAA GATAAAAGCT CGGGCATTGT GTCTTTGAAT 301 ATTCGCGGCG ACAGCGGGTT CGGGCGGGTC AATACGATGG TGGACGGCAT 351 CACGCAGACC TTTTATTCGA CTTCTACCGA TGCGGGCAGG GCAGGCGGTT 401 CATCTCAATT CGGTGCATCT GTCGACAGCA ATTTTATTGC CGGACTGGAT 451 GTCGTCAAAG GCAGCTTCAG CGGCTCGGCA GGCATCAACA GCCTTGCCGG 501 TTCGGCGAAT CTGCGGACTT TAGGCGTGGA TGACGTCGTT CAGGGCAATA 551 ATACCTACGG CCTGCTGCTA AAAGGTCTGA CCGGCACCAA TTCAACCAAA 601 GGTAATGCGA TGGCGGCGAT AGGTGCGCGC AAATGGCTGG AAAGCGGAGC 651 GTCTGTCGGT GTGCTTTACG GGCACAGCAG GCGCGGCGTG GCGCAAAATT 701 ACCGCGTGGG CGGCGGCGGG CAGCACATCG GAAATTTTGG TGAAGAATAT 751 CTGGAACGGC GCAAACAGCA ATATTTTGTA CAAGAGGGTG GTTTGAAATT 801 CAATGCCGGC AGCGGAAAAT GGGAACGGGA TTTGCAAAGG CAATACTGGA 851 AAACAAAGTG GTATAAAAAA TACGAAGACC CCCAAGAACT GCAAAAATAC 901 ATCGAAGAGC ATGATAAAAG CTGGCGGGAA AACCTGGCGC CGCAATACGA 951 CATCACCCCC ATCGATCCGT CCGGCCTGAA GCAGCAGTCG GCAGGCAATC 1001 TGTTTAAATT GGAATACGAC GGCGTATTCA ATAAATACAC GGCGCAATTT 1051 CGCGATTTAA ACACCAGAAT CGGCAGCCGC AAAATCATCA ACCGCAATTA 1101 TCAATTCAAT TACGGTTTGT CTTTGAACCC GTATACCAAC CTCAATCTGA 1151 CCGCAGCCTA CAATTCGGGC AGGCAGAAAT ATCCGAAAGG GGCGAAGTTT 1201 ACAGGCTGGG GGCTTTTAAA AGATTTTGAA ACCTACAACA ACGCGAAAAT 1251 CCTCGACCTC AACAACACCG CCACCTTCCG GCTGCCCCGC GAAACCGAGT 1301 TGCAAACCAC TTTGGGCTTC AATTATTTCC ACAACGAATA CGGCAAAAAC 1351 CGCTTTCCTG AAGAATTGGG GCTGTTTTTC GACGGTCCTG ATCAGGACAA 1401 CGGGCTTTAT TCCTATTTGG GGCGGTTTAA GGGCGATAAA GGGCTGTTGC 1451 CTCAAAAATC AACCATTGTC CAACCGGCCG GCAGCCAATA TTTCAACACG 1501 TTCTACTTCG ATGCCGCGCT CAAAAAAGAC ATTTACCGCT TAAACTACAG 1551 CACCAATGCA ATCAACTACC GTTTCGGCGG CGAATATACG GGCTATTACG 1601 GCTCGGAAAA CGAATTTAAG CGGGCATTCG GAGAAAACTC GCCGGCATAC 1651 AAGGAACATT GCGACCCGAG CTGCGGGCTT TATGAACCCG TATTGAAAAA 1701 ATACGGCAAA AAGCGCGCCA ACAACCATTC GGTCAGCATT AGTGCGGACT 1751 TCGGCGATTA TTTCATGCCG TTCGCCGGCT ATTCGCGCAC ACACCGTATG 1801 CCCAACATCC AAGAAATGTA TTTTTCCCAA ATCGGCGACT CCGGCGTTCA 1851 CACCGCCTTA AAACCAGAGC GCGCAAACAC TTGGCAATTT GGCTTCAATA 1901 CCTATAAAAA AGGATTGTTA AAACAAGATG ATATATTAGG ATTGAAACTG 1951 GTCGGCTACC GCAGCCGCAT TGACAACTAC ATCCACAACG TTTACGGGAA 2001 ATGGTGGGAT TTGAACGGGG ATATTCCGAG CTGGGTCGGC AGCACCGGGC 2051 TTGCCTACAC CATCCGACAC CGCAATTTCA AAGACAAAGT GCACAAACAC 2101 GGTTTTGAGC TGGAGCTGAA TTACGATTAT GGGCGTTTTT TCACCAACCT 2151 TTCTTACGCC TATCAAAAAA GCACGCAACC GACCAATTTC AGCGATGCGA 2201 GCGAATCGCC CAACAATGCC tccaaAGAAG ACCAACTCAA ACAAGGTTAT 2251 GGGCTGAGCA GGGTTTCCGC CCTGCCGCGA GATTACGGAC GTTTGGAAGT 2301 CGGTACGCGC TGGTTGGGCA ACAAACTGAC TTTGGGCGGC GCGAtgcGCT 2351 ATTTCGGCAA GAGCATCCGC GCGACGGCTG AAGAACGCTA TATCGACGGC 2401 ACCAACGGGG GAAATACCAG CAATGTCCGG CAACTGGGCA AGCGTTCCAT 2451 CAAACAAACC GAAACCCTTG CCCGACAGCC TTTGATTTTT GATTTTTACG 2501 CCGCTTACGA GCCGAAGAAA AACCTTATTT TCCGCGCCGA AGTCAAAAAC 2551 CTGTTCGACA GGCGTTATAT CGATCCGCTC GATGCGGGCA ATGATGCGGC 2601 AACGCAGCGT TATTACAGCT CGTTCGACCC GAAAGACAAG GACGAAGACG 2651 TAACGTGTAA TGCTGATAAA ACGTTGTGCA ACGGCAAATA CGGCGGCACA 2701 AGCAAAAGCG TATTGACCAA TTTCGCACGC GGACGCACCT TCTTGATGAC 2751 GATGAGCTAC AAGTTTTAA This coπesponds to the .amino acid sequence <SEQ ID 884; ORF133ng-l>: 1 MRSSFRLKPI CFYLMGVMLY HHSYAEDAGR AGSEAQIQVL EDVHVKAKRV 51 PKDKKVFTDA RAVSTRQDVF KSGENLDNIV RSIPGAFTQQ DKSSGIVSLN 101 IRGDSGFGRV NTMVDGITQT FYSTSTDAGR AGGSSQFGAS VDSNFIAGLD 151 VVKGSFSGSA GINSLAGSAN LRTLGVDDW QGNNTYGLLL KGLTGTNSTK 201 GNAMAAIGAR KWLESGASVG VLYGHSRRGV AQNYRVGGGG QHIGNFGEEY 251 LERRKQQYFV QEGGLKFNAG SGKWERDLQR QYWKTKWYKK YEDPQELQKY 301 IEEHDKSWRE NLAPQYDITP IDPSGLKQQS AGNLFKLEYD GVFNKYTAQF 351 RDLNTRIGSR KIINRNYQFN YGLSLNPYTN LNLTAAYNSG RQKYPKGAKF 401 TGWGLLKDFE TYNNAKILDL NNTATFRLPR ETELQTTLGF NYFHNEYGKN 451 RFPEELGLFF DGPDQDNGLY SYLGRFKGDK GLLPQKSTIV QPAGSQYFNT 501 FYFDAALKKD IYRLNYSTNA INYRFGGEYT GYYGSENEFK RAFGENSPAY 551 KEHCDPSCGL YEPVLKKYGK KRANNHSVSI SADFGDYFMP FAGYSRTHRM 601 PNIQEMYFSQ IGDSGVHTAL KPERANTWQF GFNTYKKGLL KQDDILGLKL 651 VGYRSRIDNY IHNVYGKWWD LNGDIPSWVG STGLAYTIRH RNFKDKVHKH 701 GFELELNYDY GRFFTNLSYA YQKSTQPTNF SDASESPNNA SKEDQLKQGY 751 GLSRVSALPR DYGRLEVGTR WLGNKLTLGG AMRYFGKSIR ATAEERYIDG 801 TNGGNTSNVR QLGKRSIKQT ETLARQPLIF DFYAAYEPKK NLIFRAEVKN 851 LFDRRYIDPL DAGNDAATQR YYSSFDPKDK DEDVTCNADK TLCNGKYGGT 901 SKSVLTNFAR GRTFLMTMSY KF* ORF133ng-l and ORF133-1 show 96.2% identity in 889 aa overlap: 10 20 30 40 50 60 orfl33ng-l.pep SFRLKPICFYLMGVMLYHHSYAEDAGRAGSEAQIQVLEDVHVKAKRVPKDKKVFTDARAV I I I II I I I I I I II II I I I I II II I II I I I I orf133-1 EAQIQVLEDVHVKAKRVPKDKKVFTDARAV 10 20 30 70 80 90 100 110 120 orf133ng-l . ep STRQDVFKSGENLDNIVRSIPGAFTQQDKSSGIVSLNIRGDSGFGRVNTMVDGITQTFYS I II I M I I M I I II II II I II I II II II II I I I I I I I I II I II II II I I II II I I II I II orf133-1 STRQDIFKSSENLDNIVRSIPGAFTQQDKSSGIVSLNIRGDSGFGRVNTMVDGITQTFYS 40 50 60 70 80 90 130 140 150 160 170 180 orfl33ng-l.pep TSTDAGRAGGSSQFGASVDSNFIAGLDWKGSFSGSAGINSLAGSANLRTLGVDDWQGN II I I I I I II I I I I I I II I I I I M I I II I I I I I I I I I I I I I I I I I I I II I I I I I I I I I I II orf133-1 TSTDAGRAGGSSQFGASVDSNFIAGLDWKGSFSGSAGINSLAGSANLRTLGVDDWQGN 100 110 120 130 140 150 190 200 210 220 230 240 orf133ng-l . pep NTYGLLLKGLTGTNSTKGNAMAAIGARKWLESGASVGVLYGHSRRGVAQNYRVGGGGQHI II I I I II I I I II I I II I I II I I II I I I I II I I I M II II I I I I IM I I II I I I II I II I I orf133-1 NTYGLLLKGLTGTNSTKGNAMAAIGARKWLESGASVGVLYGHSRRSVAQNYRVGGGGQHI 160 170 180 190 200 210 250 260 270 280 290 300 orf133ng-l . pep GNFGEEYLERRKQQYFVQEGGLKFNAGSGKWERDLQRQYWKTKWYKKYEDPQELQKYIEE I II I I I I II I II : II I II I: I II I : I II I II I I II I II I I M: II I I I I I II orf133-1 GNFGAEYLERRKQRYFVQEGALKFNSDSGKWERDLQRQQWKYKPYKNYNN-QELQKYIEE 220 230 240 250 260 310 320 330 340 350 360 orfl33ng-l.pep HDKSWRENLAPQYDITPIDPSGLKQQSAGNLFKLEYDGVFNKYTAQFRDLNTRIGSRKII I II I II I II I II I I II I II I: I I II II I II I I II I I I I I I I II I I I I II I I : I I I I I II orf133-1 HDKSWRENLXPQYDITPIDPSSLKQQSAGNLFKLEYDGVFNKYTAQFRDLNTKIGSRKII 270 280 290 300 310 320 370 380 390 400 410 420 orfl33ng-l.pep NRNYQFNYGLSLNPYTNLNLTAAYNSGRQKYPKGAKFTGWGLLKDFETYNNAKILDLNNT I II I I I II I I I II I I II I II I I I I II I I I I I II I M I I II I II I I I I II II I II I II I I I orf133-1 NRNYQFNYGLSLNPYTNLNLTAAYNSGRQKYPKGSKFTGWGLLKDFETYNNAKILDLNNT 330 340 350 360 370 380 430 440 450 460 470 480 orf133ng-l . pep ATFRLPRETELQTTLGFNYFHNEYGKNRFPEELGLFFDGPDQDNGLYSYLGRFKGDKGLL I I I I II I II I II I I II I II I I I I II II II I I I I I I I I I I II I II II I II I I I I I I I I I I I orf133-1 ATFRLPRETELQTTLGFNYFHNEYGKNRFPEELGLFFDGPDQDNGLYSYLGRFKGDKGLL 390 400 410 420 430 440 490 500 510 520 530 540 orfl33ng-l.pep PQKSTIVQPAGSQYFNTFYFDAALKKDIYRLNYSTNAINYRFGGEYTGYYGSENEFKRAF I II I II I I I II I II I I I II I II II I II II I II I II I ::: II II I II I II I I I :: I II I I I orfl33-1 PQKSTIVQPAGSQYFNTFYFDAALKKDIYRLNYSTNTVGYRFGGEYTGYYGSDDEFKRAF 450 460 470 480 490 500 550 560 570 580 590 600 orf133ng-l .pep GENSPAYKEHCDPSCGLYEPVLKKYGKKRANNHSVSISADFGDYFMPFAGYSRTHRMPNI I II I I: IM I I : I I I : I I II I II I I I I I I I I I II I I I I I I I I I II I I I : I II I I I I I I I orf133-1 GENSPTYKKHCNRSCGIYEPVLKKYGKKRANNHSVSISADFGDYFMPFASYSRTHRMPNI 510 520 530 540 550 560 610 620 630 640 650 660 orf133ng-l . pep QEMYFSQIGDSGVHTALKPERANTWQFGFNTYKKGLLKQDDILGLKLVGYRSRIDNYIHN II I I I I I I II I II I I II I I II I I I I I I I I I II I I I I I I I II I I I I I I I I I I II I I I I I I orf133-1 QEMYFSQIGDSGVHTALKPERANTWQFGFNTYKKGLLKQDDTLGLKLVGYRSRIDNYIHN 570 580 590 600 610 620 670 680 690 700 710 720 orf133ng-l . pep VYGKWWDLNGDIPSWVGSTGLAYTIRHRNFKDKVHKHGFELELNYDYGRFFTNLSYAYQK I II I I I II I II I II I I : II I I I I II : I I II I I II I I I I I I II I I I II I II I I I I I I I I II orf133-1 VYGKWWDLNGDIPSWVSSTGLAYTIQHRNFKDKVHKHGFELELNYDYGRFFTNLSYAYQK 630 640 650 660 670 680 730 740 750 760 770 780 orf133ng-l . pep STQPTNFSDASESPNNASKEDQLKQGYGLSRVSALPRDYGRLEVGTRWLGNKLTLGGAMR I I II I I I I I I II I I II II II I I I I I I I I I I I I I II I I I I I I II II I II I II I II II II I I orf133-1 STQPTNFSDASESPNNASKEDQLKQGYGLSRVSALPRDYGRLEVGTRWLGNKLTLGGAMR 690 700 710 720 730 740 790 800 810 820 830 840 orfl33ng-l.pep YFGKSIRATAEERYIDGTNGGNTSNVRQLGKRSIKQTETLARQPLIFDFYAAYEPKKNLI I II I II I II I I I I I II II II II I I I I I I II II I I II II I I I I I I I II I I II I II II II I orf133-1 YFGKSIRATAEERYIDGTNGGNTSNFRQLGKRSIKQTETLARQPLIFDFYAAYEPKKNLI 750 760 770 780 790 800 850 860 870 880 890 900 orf133ng-l .pep FRAEVKNLFDRRYIDPLDAGNDAATQRYYSSFDPKDKDEDVTCNADKTLCNGKYGGTSKS I I I I II I II I I I I II II I I II I I I I I I I I I I I II II II I I II II I I I I II I II I I I I I II orf133-1 FRAEVKNLFDRRYIDPLDAGNDAATQRYYSSFDPKDKDEDVTCNADKTLCNGKYGGTSKS 810 820 830 840 850 860 910 920 orfl33ng-l .pep VLTNFARGRTFLMTMSYKFX I II I II I I I I II I I I II I I I orfl33-l VLTNFARGRTFLMTMSYKFX 870 880 ition, ORF 133ng- 1 is homologous to a TonB-dependent receptor in H. influenzae: sp|P45114 |YC17_HAEIN PROBABLE TONB-DEPENDENT RECEPTOR HI1217 PRECURSOR >gι 11075372 |pιr||G64110 transferrin binding protein 1 precursor (tbpl) homolog - Haemophilus influenzae (strain Rd KW20) >gι 11574147 (U32801) transferrin binding protein 1 precursor (tbpl) [Haemophilus influenzae] Length = 913 Score = 930 bits (2377), Expect = 0.0 Identities = 476/921 (51%), Positives = 619/921 (66%), Gaps = 72/921 (7%) Query: 38 QVLEDVHVKAKRVPKDKKVFTDARAVSTRQDVFKSGENLDNIVRSIPGAFTQQDKSSGIV 97 + L + V K + DKK FT+A+A STR++VFK + +D ++RSIPGAFTQQDK SG+V Sb j ct: 29 ETLGQIDWEKVISNDKKPFTEAKAKSTRENVFKETQTIDQVIRSIPGAFTQQDKGSGW 88 Query: 98 SLNIRGDSGFGRVNTMVDGITQTFYSTSTDAGRAGGSSQFGASVDSNFIAGLDWKGSFS 157 S+NIRG++G GRVNTMVDG+TQTFYST+ D+G++GGSSQFGA++D NFIAG+DV K +FS Sbjct: 89 SVNIRGENGLGRVNTMVDGVTQTFYSTALDSGQSGGSSQFGAAIDPNFIAGVDVNKSNFS 148 Query: 158 GSAGINSLAGSANLRTLGVDDWQXXXXXXXXXXXXXXXXXXXXXAMAAIGARKWLESGA 217 G++GIN+LAGSAN RTLGV+DV+ M RKWL++G Sbjct: 149 GASGINALAGSANFRTLGVNDVITDDKPFGIILKGMTGSNATKSNFMTMAAGRKWLDNGG 208 Query: 218 SVGVLYGHSRRGVAQNYRVGGGGQHIGNFGEEYLERRKQQYFVQEGGLKFNAGSGKWERD 277 VGV+YG+S+R V+Q+YR+ GGG+ + + G++ L + K+ YF + G N G+W D Sbjct: 209 YVGWYGYSQREVSQDYRI-GGGERLASLGQDILAKEKEAYF-RNAGYILNP-EGQWTPD 265 Query: 278 LQRQYWK TKWY KKYEDPQELQK YIEE 303 L +++W +Y KK +D ++LQK IEE Sbjct: 266 LSKKHWSCNKPDYQKNGDCSYYRIGSAAKTRREILQELLTNGKKPKDIEKLQKGNDGIEE 325 Query: 304 HDKSWRENLAPQYDITPIDPSGLKQQSAGNLFKLEYDGVFNKYTAQFRDLNTRIGSRKII 363 DKS+ N QY + PI+P L+ +S +L K EY AQ R L+ +IGSRKI Sbjct: 326 TDKSFERN-KDQYSVAPIEPGSLQSRSRSHLLKFEYGDDHQNLGAQLRTLDNKIGSRKIE 384 Query: 364 NRNYQFNYGLSLNPYTNLNLTAAYNSGRQKYPKGAKFTGWGLLKDFETYNNAKILDLNNT 423 NRNYQ NY + N Y +LNL AA+N G+ YPKG F GW + T N A I+D+NN+ Sbjct: 385 NRNYQVNYNFNNNSYLDLNLMAAHNIGKTIYPKGGFFAGWQVADKLITKNVANIVDINNS 444 Query: 424 ATFRLPRETELQTTLGFNYFHNEYGKNRFPEELGLFFDGPDQDNGLYSY— GRFKGDKG 481 TF LP+E +L+TTLGFNYF NEY KNRFPEEL LF++ D GLYS+ GR+ G K Sbjct: 445 HTFLLPKEIDLKTTLGFNYFTNEYSKNRFPEELSLFYNDASHDQGLYSHSKRGRYSGTKS 504 Query: 482 LLPQKSTIVQPAGSQYFNTFYFDAALKKDIYRLNYSTNAINYRFGGEYTGYYGSENEFKR 541 LLPQ+S I+QP+G Q F T YFD AL K IY LNYS N +Y F GEY GY Sbjct: 505 LLPQRSVILQPSGKQKFKTVYFDTALSKGIYHLNYSVNFTHYAFNGEYVGY 555 Query: 542 AFGENSPAYKEHCDPSCGLYEPVLKKYGKKRANNHSVSISADFGDYFMPFAGYSRTHRMP 601 EN+ + + EP+L K G K+A NHS ++SA+ DYFMPF YSRTHRMP Sbjct: 556 ENTAGQQ INEPILHKSGHKKAFNHSATLSAELSDYFMPFFTYSRTHRMP 604 Query: 602 NIQEMYFSQIGDSGVHTALKPERANTWQFGFNTYKKGLLKQDDILGLKLVGYRSRIDNYI 661 NIQEM+FSQ+ ++GV+TALKPE+++T+Q GFNTYKKGL QDD+LG+KLVGYRS I NYI Sbjct: 605 NIQEMFFSQVSNAGVNTALKPEQSDTYQLGFNTYKKGLFTQDDVLGVKLVGYRSFIKNYI 664 Query: 662 HNVYGKWWDLNGDIPSWVGSTGLAYTIRHRNFKDKVHKHGFELELNYDYGRFFTNLSYAY 721 HNVYG WW +P+W S G YTI H+N+K V K G ELE+NYD GRFF N+SYAY Sbjct: 665 HNVYGVWW—RDGMPTWAESNGFKYTIAHQNYKPIVKKSGVELEINYDMGRFFANVSYAY 722 Query: 722 QKSTQPTNFSDASESPNNASKEDQLKQGYGLSRVSALPRDYGRLEVGTRWLGNKLTLGGA 781 Q++ QPTN++DAS PNNAS+ED LKQGYGLSRVS LP+DYGRLE+GTRW KLTLG A Sbjct: 723 QRTNQPTNYADASPRPNNASQEDILKQGYGLSRVSMLPKDYGRLELGTRWFDQKLTLGLA 782 Query: 782 MRYFGKSIRATAEERYIDGTNGGNTSNVRQLGKRSIKQTETLARQPLIFDFYAAYEPKKN 841 RY+GKS RAT EE YI+G+ + +R+ ++K+TE + +QP+I D + +YEP K+ Sbjct: 783 ARYYGKSKRATIEEEYINGSR-FKKNTLRRENYYAVKKTEDIKKQPIILDLHVSYEPIKD 841 Query: 842 LIFRAEVKNLFDRRYIDPLDAGNDAATQRYYSSFDPKDKDEDVTCNADKTLCNGKYGGTS 901 LI +AEV+NL D+RY+DPLDAGNDAA+QRYYSS + + C D + C GG+ Sbjct: 842 LIIKAEVQNLLDKRYVDPLDAGNDAASQRYYSSL NNSIECAQDSSAC GGSD 892 Query: 902 KSVLTNFARGRTFLMTMSYKF 922 K+VL NFARGRT++++++YKF Sbjct: 893 KTVLYNFARGRTYILSLNYKF 913 The underlined motif in the gonococcal protein (also present in the meningococcal protein) is predicted to be an ATP/GTP-binding site motif A (P-loop), and the analysis suggests that these proteins from N. meningitidis and N.gonorrhoeae, and their epitopes, could be useful antigens for vaccines or diagnostics, or for raising antibodies. Example 104 The following partial DNA sequence was identified in N. meningitidis <SEQ ID 885> 1 ATGAACCTGA TTTCACGTTA CATCATCCGT CAAATGGCGG TTATGGCGGT 51 TTACGCGCTC CTTGCCTTCC TCGCTTTGTA CAGCTTTTTT GAAATCCTGT 101 ACGAAACCGG CAACCTCGGC AAAGGCAGTT ACGGCATATG GGAAATGCTG 151 GGCTACACCG CCCTCAAAAT GCCCGCCCGC GCCTACGAAC TGATTCCCCT 201 CGCCGTCCTT ATCGGCGGAC TGGTCTCCCT CAGCCAGCTT GCCGCCGGCA 251 GCGAACTGAC CGTCATCAAA GCCAGCGGCA TGAGCACCAA AAAGCTGCTG 301 TTGATTCTGT CGCAGTTCGG TTTTATTTTT GCTATTGCCA CCGTCGCGCT 351 CGGCGAATGG GTTGCGCCCA CACTGAGCCA AAAAGCCGAA AACATCAAAG 401 CCGCCGCCAT CAACGGCAAA ATCAGCACCG GCAATACCGG CCTTTGGCTG 451 AAAGAAAAAA ACAGCGTGAT CAATGTGCGC GAAATGTTGC CCGACCAT .. This coπesponds to the amino acid sequence <SEQ ID 886; ORFl 12>: 1 MNLISRYIIR QMAVMAVYAL LAFLALYSFF EILYETGNLG KGSYGIWEML 51 GYTALKMPAR AYELIPLAVL IGGLVSLSQL AAGSELTVIK ASGMSTKKLL 101 LILSQFGFIF AIATVALGEW VAPTLSQKAE NIKAAAINGK ISTGNTGLWL 151 KEKNSVINVR EMLPDH... Further work revealed further partal nucleotide sequence <SEQ ED 887>: 1 ATGAACCTGA TTTCACGTTA CATCATCCGT CAAATGGCGG TTATGGCGGT 51 TTACGCGCTC CTTGCCTTCC TCGCTTTGTA CAGCTTTTTT GAAATCCTGT 101 ACGAAACCGG CAACCTCGGC AAAGGCAGTT ACGGCATATG GGAAATGCTG 151 gGCTACACCG CCCTCAAAAT GCCCGCCCGC GCCTACGAAC TGATTCCCCT 201 CGCCGTCCTT ATCGGCGGAC TGGTCTCCCT CAGCCAGCTT GCCGCCGGCA 251 GCGAACTGAC CGTCATCAAA GCCAGCGGCA TGAGCACCAA AAAGCTGCTG 301 TTGATTCTGT CGCAGTTCGG TTTTATTTTT GCTATTGCCA CCGTCGCGCT 351 CGGCGAATGG GTTGCGCCCA CACTGAGCCA AAAAGCCGAA AACATCAAAG 401 CCGCCGCCAT CAACGGCAAA ATCAGCACCG GCAATACCGG CCTTTGGCTG 451 AAAGAAAAAA ACAGCrTkAT CAATGTGCGC GAAATGTTGC CCGACCATAC 501 GCTTTTGGGC ATCAAAATTT GGGCGCGCAA CGATAAAAAC GAATTGGCAG 551 AGGCAGTGGA AGCCGATTCC GCCGTTTTGA ACAGCGACGG CAGTTGGCAG 601 TTGAAAAACA TCCGCCGCAG CACGCTTGGC GAAGACAAAG TCGAGGTCTC 651 TATTGCGGCT GAAGAAAACT GGCCGATTTC CGTCAAACGC AACCTGATGG 701 ACGTATTGCT CGTCAAACCC GACCAAATGT CCGTCGGCGA ACTGACCACC 751 TACATCCGCC ACCTCCAAAA CAACAGCCAA AACACCCGAA TCTACGCCAT 801 CGCATGGTGG CGCAAATTGG TTTACCCCGC CGCAGCCTGG GTGATGGCGC 851 TCGTCGCCTT TGCCTTTACC CCGCAAACCA CCCGCCACGG CAATATGGGC 901 TTAAAACTCT TCGGCGGCAT CTGTsTCGGA TTGCTGTTCC ACCTTGCCGG 951 ACGGCTCTTT GGGTTTACCA GCCAACTCGG... This coπesponds to the amino acid sequence <SEQ ED 888; ORFl 12-1>: 1 MNLISRYIIR QMAVMAVYAL LAFLALYSFF EILYETGNLG KGSYGIWEML 51 GYTALKMPAR AYELIPLAVL IGGLVSLSQL AAGSELTVIK ASGMSTKKLL 101 LILSQFGFIF AIATVALGEW VAPTLSQKAE NIKAAAINGK ISTGNTGLWL 151 KEKNSXINVR EMLPDHTLLG IKIWARNDKN ELAEAVEADS AVLNSDGSWQ 201 LKNIRRSTLG EDKVEVSIAA EENWPISVKR NLMDVLLVKP DQMSVGELTT 251 YIRHLQNNSQ NTRIYAIAWW RKLVYPAAAW VMALVAFAFT PQTTRHGNMG 301 LKLFGGICXG LLFHLAGRLF GFTSQL... Computer analysis of this amino acid sequence predicts two transmembrane domains and gave the following results: Homologv with a predicted ORF from N.meninsitidis (sfrain A) ORFl 12 shows 96.4% identity over a 166aa overlap with an ORF (ORFl 12a) from strain A of N. meningitidis: 10 20 30 40 50 60 orf112. pep MΝLISRYIIRQMAVMAVYALLAFLALYSFFEILYETGΝLGKGSYGIWEMLGYTALKMPAR I I I I I I II I II I I I II II I I II I I I I I I I II I II I II I I I I I I I I II I I I I II I I I II orf112a MΝLISRYIIRQMAVMAVYALLAFLALYSFFEILYETGΝLGKGSYGIWEMXGYTALKMXAR 10 20 30 40 50 60 70 80 90 100 110 120 orf112. pep AYELIPLAVLIGGLVSLSQLAAGSELTVIKASGMSTKKLLLILSQFGFIFAIATVALGEW II I I : I II I I I I II I I I II I II I II : I I I I II I II II I I I I I I I I II I I I I II I I I I II orf112a AYELMPLAVLIGGLVSXSQLAAGSELXVIKASGMSTKKLLLILSQFGFIFAIATVALGEW 70 80 90 100 110 120 130 140 150 160 orf112.pep VAPTLSQKAEΝIKAAAIΝGKISTGΝTGLWLKEKΝSVIΝVREMLPDH I I I I I I II I I I I II I I I I I II I I I II I I I I II II I: I I I I I II I I I orf112a VAPTLSQKAEΝIKAAAIΝGKISTGΝTGLWLKEKΝSIIΝVREMLPDHTLLGIKIWARΝDKΝ 130 140 150 160 170 180 orfll2a ELAEAVEADSAVLΝSDGSWQLKΝIRRSTLGEDKVEVSIAAEEXWPISVKRΝLMDVLLVKP 190 200 210 220 230 240 The ORFl 12a nucleotide sequence <SEQ ID 889> is: 1 ATGAACCTGA TTTCACGTTA CATCATCCGT CAAATGGCGG TTATGGCGGT 51 TTACGCGCTC CTTGCCTTCC TCGCTTTGTA CAGCTTTTTT GAAATCCTGT 101 ACGAAACCGG CAACCTCGGC AAAGGCAGTT ACGGCATATG GGAAATGΝTG 151 GGΝTACACCG CCCTCAAAAT GΝCCGCCCGC GCCTACGAAC TGATGCCCCT 201 CGCCGTCCTT ATCGGCGGAC TGGTCTCTΝT CAGCCAGCTT GCCGCCGGCA 251 GCGAACTGAΝ CGTCATCAAA GCCAGCGGCA TGAGCACCAA AAAGCTGCTG 301 TTGATTCTGT CGCAGTTCGG TTTTATTTTT GCTATTGCCA CCGTCGCGCT 351 CGGCGAATGG GTTGCGCCCA CACTGAGCCA AAAAGCCGAA AACATCAAAG 401 CCGCGGCCAT CAACGGCAAA ATCAGTACCG GCAATACCGG CCTTTGGCTG 451 AAAGAAAAAA ACAGCATTAT CAATGTGCGC GAAATGTTGC CCGACCATAC 501 CCTGCTGGGC ATTAAAATCT GGGCCCGCAA CGATAAAAAC GAACTGGCAG 551 AGGCAGTGGA AGCCGATTCC GCCGTTTTGA ACAGCGACGG CAGTTGGCAG 601 TTGAAAAACA TCCGCCGCAG CACGCTTGGC GAAGACAAAG TCGAGGTCTC 651 TATTGCGGCT GAAGAAAAΝT GGCCGATTTC CGTCAAACGC AACCTGATGG 701 ACGTATTGCT CGTCAAACCC GACCAAATGT CCGTCGGCGA ACTGACCACC 751 TACATCCGCC ACCTCCAAAΝ ΝΝACAGCCAA AACACCCGAA TCTACGCCAT 801 CGCATGGTGG CGCAAATTGG TTTACCCCGC CGCAGCCTGG GTGATGGCGC 851 TCGTCGCCTT TGCCTTTACC CCGCAAACCA CCCGCCACGG CAATATGGGC 901 TTAAAAΝTCT TCGGCGGCAT CTGTCTCGGA TTGCTGTTCC ACCTTGCCGG 951 ΝCGGCTCTTC ΝGGTTTACCA GCCAACTCTA CGGCATCCCG CCCTTCCTCG 1001 ΝCGGCGCACT ACCTACCATA GCCTTCGCCT TGCTCGCCGT TTGGCTGATA 1051 CGCAAACAGG AAAAACGCTA A This encodes a protein having the amino acid sequence <SEQ ID 890>: 1 MNLISRYIIR QMAVMAVYAL LAFLALYSFF EILYETGNLG KGSYGIWEMX 51 GYTALKMXAR AYELMPLAVL IGGLVSXSQL AAGSELXVIK ASGMSTKKLL 101 LILSQFGFIF AIATVALGEW VAPTLSQKAE NIKAAAINGK ISTGNTGLWL 151 KEKNSIINVR EMLPDHTLLG IKIWARNDKN ELAEAVEADS AVLNSDGSWQ 201 LKNIRRSTLG EDKVEVSIAA EEXWPISVKR NLMDVLLVKP DQMSVGELTT 251 YIRHLQXXSQ NTRIYAIAWW RKLVYPAAAW VMALVAFAFT PQTTRHGNMG 301 LKXFGGICLG LLFHLAGRLF XFTSQLYGIP PFLXGALPTI AFALLAVWLI 351 RKQEKR* ORFl 12a and ORFl 12-1 show 96.3% identity in 326 aa overlap: orf112a . pep MNLISRYIIRQMAVMAVYALLAFLALYSFFEILYETGNLGKGSYGIWEMXGYTALKMXAR I I I I I II I I I I II I I I II I II I I I I I I I I I I I I II I I II I II I I II I I I II I I I I I II orf112-1 MNLISRYIIRQMAVMAVYALLAFLALYSFFEILYETGNLGKGSYGIWEMLGYTALKMPAR orf112a. pep AYELMPLAVLIGGLVSXSQLAAGSELXVIKASGMSTKKLLLILSQFGFIFAIATVALGEW I I I I : I I II I I I II I I I I II I II I I : I I I I II II I I I I I I I II I I I I I II II I II II I I orfl12-1 AYELIPLAVLIGGLVSLSQLAAGSELTVIKASGMSTKKLLLILSQFGFIFAIATVALGEW orf112a . pep VAPTLSQKAENIKAAAINGKISTGNTGLWLKEKNSIINVREMLPDHTLLGIKIWARNDKN II II II II I II II II I II I II I I I I I I I I I I II II I I I I I I I I I I I I II I I II II II I I orf112-1 VAPTLSQKAENIKAAAINGKISTGNTGLWLKEKNSXINVREMLPDHTLLGIKIWARNDKN orf112a . pep ELAEAVEADSAVLNSDGSWQLKNIRRSTLGEDKVEVSIAAEEXWPIΞVKRNLMDVLLVKP I I II I I II I I I II II I I I II I II I I I II I I I I I II I I I I II I II I I I II I I I II I I II I orf112-1 ELAEAVEADSAVLNSDGSWQLKNIRRSTLGEDKVEVSIAAEENWPISVKRNLMDVLLVKP orf112a. pep DQMSVGELTTYIRHLQXXSQNTRIYAIAWWRKLVYPAAAWVMALVAFAFTPQTTRHGNMG I II I I I I I II I I I I I I II I I I II I II II II I II I I I I I I I I I I I I I II I I II II II I I orfl12-1 DQMSVGELTTYIRHLQNNSQNTRIYAIAWWRKLVYPAAAWVMALVAFAFTPQTTRHGNMG orf112a . pep LKXFGGICLGLLFHLAGRLFXFTSQLYGIPPFLXGALPTIAFALLAVWLIRKQEKRX II II II I I II I I I I I I I I II II I orf112-1 LKLFGGICXGLLFHLAGRLFGFTSQL Homology with a predicted ORF from Nεonorrhoeae ORFl 12 shows 95.8% identity over 166aa overlap with a predicted ORF (ORF112ng) from N. gonorrhoeae: orf112.pep MNLISRYIIRQMAVMAVYALLAFLALYSFFEILYETGNLGKGSYGIWEMLGYTALKMPAR 60 I II II II I I I II II I II I II I II I I I I II I I I II I II I I I I II II II I II I I II I I I II I orfll2ng MNLISRYIIRQMAVMAVYALLAFLALYSFFEILYETGNLGKGSYGIWEMLGYTALKMPAR 60 orf112.pep AYELIPLAVLIGGLVSLSQLAAGSELTVIKASGMSTKKLLLILSQFGFIFAIATVALGEW 120 II I I : I I I I I I I I I : I I II I II I I I I : I I II II I I I I I I I I I I II II I II I I I : I I I I I I orfll2ng AYELMPLAVLIGGLASLSQLAAGSELAVIKASGMSTKKLLLILSQFGFIFAIAAVALGEW 120 orf112. pep VAPTLSQKAENIKAAAINGKISTGNTGLWLKEKNSVINVREMLPDH 166 I I I I I I II II I I I I II I I I I I I I II II I II I II: I : I I I I I I I I I orfll2ng VAPTLSQKAENIKAAAINGKISTGNTGLWLKEKTSIINVRGMLPDHTLLGIKIWARNDKN 180 The complete length ORFl 12ng nucleotide sequence <SEQ ID 891 > is: 1 ATGAACCTGA TTTCACGTTA CATCATCCGC CAAATGGCGG TTATGGCGGT 51 TTACGCGCTC CTTGCCTTCC TCGCTTTGTA CAGCTTTTTT GAAATCCTGT 101 ACGAAACCGG CAACCTCGGC AAAGGCAGTT ACGGCATATG GGAAATGCTG 151 GGCTACACCG CCCTCAAAAT GCCCGCCCGC GCCTACGAAC TCATGCCCCT 201 CGCCGTCCTC ATCGGCGGAC TGGCCTCTCT CAGCCAGCTT GCCGCCGGCA 251 GCGAACTGGC CGTCATCAAA GCCAGCGGCA TGAGCACCAA AAAGCTGCTG 301 TTGATTCTGT CTCAGTTCGG TTTTATTTTT GCTATTGCCG CCGTCGCGCT 351 CGGCGAATGG GTTGCGCCCA CGCTGAGCCA AAAAGCCGAA AACATCAAag 401 cCGCCGCCAt taacggCAAA ATCAGCAccg gcAATACCGG CCTTTggcTG 451 AAAGAAAAAa ccAGCATTAT CAATGTGcGc GGAATGTTGC CCGACCATAC 501 GCTTTTGGGC ATCAAAATTT GGGCGCGCAA CGATAAAAAC GAATTGGCAG 551 AGGCAGTGGA AGCCGATTCC GCCGTTTTGA ACAGCGACGG CAGCTGGCAG 601 TTGAAAAACA TCCGCCGCAG CATCATGGGT ACAGACAAAA TCGAAACATC 651 cgCCGCCGCC GAAGAAACTT gGCCGATTGC CGTCAGACGC AACCTGATGG 701 ACGTATTGCT CGTCAAGCCC GACCAAATGT CCGTCGGCGA GCTGACCACC 751 TACATCCGCC ACCTCCAAAA CAACAGCCAA AACACCCAAA TCTACGCCAT 801 CGCATGGTGG CGTAAACTCG TTTACCCCGT CGCCGCATGG GTCATGGCGC 851 TCGTTGCCTT CGCCTTTACG CCGCAAACCA CGCGCCACGG CAATATGGGC 901 TTAAAACTCT TCGGCGGCAT CTGTCTCGGA TTGCTGTTCC ACCTTGCCGG 951 CAGGCTCTTC GGGTTTACCA GCCAACTCTA CGGCACCCCA CCCTTCCTCG 1001 CCGGCGCACT GCCTACCATA GCCTTCGCCT TGCTCGCTGT TTGGCTGATA 1051 CGCAAACAGG AAAAACGTTG A This encodes a protein having amino acid sequence <SEQ ID 892>: 1 MNLISRYIIR QMAVMAVYAL LAFLALYSFF EILYETGNLG KGSYGIWEML 51 GYTALKMPAR AYELMPLAVL IGGLASLSQL AAGSELAVIK ASGMSTKKLL 101 LILSQFGFIF AIAAVALGEW VAPTLSQKAE NIKAAAINGK ISTGNTGLWL 151 KEKTSIINVR GMLPDHTLLG IKIWARNDKN ELAEAVEADS AVLNSDGSWQ 201 LKNIRRSIMG TDKIETSAAA EETWPIAVRR NLMDVLLVKP DQMSVGELTT 251 YIRHLQNNSQ NTQIYAIAWW RKLVYPVAAW VMALVAFAFT PQTTRHGNMG 301 LKLFGGICLG LLFHLAGRLF GFTSQLYGTP PFLAGALPTI AFALLAVWLI 351 RKQEKR* ORFl 12ng and ORFl 12-1 show 94.2% identity in 326 aa overlap: 10 20 30 40 50 60 orfll2ng MNLISRYIIRQMAVMAVYALLAFLALYSFFEILYETGNLGKGSYGIWEMLGYTALKMPAR I I I I I I I I II II II I II I II I I I I I I I II I II I II I I I I II II II I I II I I II II II I I I orf112-1 MNLISRYIIRQMAVMAVYALLAFLALYSFFEILYETGNLGKGSYGIWEMLGYTALKMPAR 10 20 30 40 50 60 70 80 90 100 110 120 orfll2ng AYELMPLAVLIGGLASLSQLAAGSELAVIKASGMSTKKLLLILSQFGFIFAIAAVALGEW I I I I: II II I I I I I : I II II I I I I II I I I I I II II I I II II II I II I II I I: I I I II I orfl12-1 AYELIPLAVLIGGLVSLSQLAAGSELTVIKASGMSTKKLLLILSQFGFIFAIATVALGEW 70 80 90 100 110 120 130 140 150 160 170 180 orfll2ng VAPTLSQKAENIKAAAINGKISTGNTGLWLKEKTSIINVRGMLPDHTLLGIKIWARNDKN I I I I I I II II I II I I I II I I I II II II I I I I I I: I I I I I I I I I II I I I I I I I I I I I I I orf112-1 VAPTLSQKAENIKAAAINGKISTGNTGLWLKEKNSXINVREMLPDHTLLGIKIWARNDKN 130 140 150 160 170 180 190 200 210 220 230 240 orfll2ng ELAEAVEADSAVLNSDGSWQLKNIRRSIMGTDKIETSAAAEETWPIAVRRNLMDVLLVKP II I II I I I II II I I I I I I I I I I I II II : I MMM I II I : II I : I : I II I I I I I I I I orf112-1 ELAEAVEADSAVLNSDGSWQLKNIRRSTLGEDKVEVSIAAEENWPISVKRNLMDVLLVKP 190 200 210 220 230 240 250 260 270 280 290 300 orfll2ng DQMSVGELTTYIRHLQNNSQNTQIYAIAWWRKLVYPVAAWVMALVAFAFTPQTTRHGNMG II I I I II I I I II I I I II II I I I : II II I I I I I I II I : I II I I I I I I I I I I II I I I I I I II orfl12-1 DQMSVGELTTYIRHLQNNSQNTRIYAIAWWRKLVYPAAAWVMALVAFAFTPQTTRHGNMG 250 260 270 280 290 300 310 320 330 340 350 orfll2ng LKLFGGICLGLLFHLAGRLFGFTSQLYGTPPFLAGALPTIAFALLAVWLIRKQEKRX I II I II II I I I I II II II I II I II I orf112-1 LKLFGGICXGLLFHLAGRLFGFTSQL 310 320 This analysis suggests that these proteins from N. meningitidis .and N.gonorrhoeae, and their epitopes, could be useful antigens for vaccines or diagnostics, or for raising antibodies. It will be appreciated that the invention has been described by means of example only, and that modifications may be made whilst remaining within the spirit and scope of the invention. TABLE I - PCR primers ORF Primer Sequence Restriction sites ORF l Forward CGCGGATCCGCTAGC-GGACACACTTATTTCGG BamHI-Nhel Reverse CCCGCTCGAG-CCAGCGGTAGCCTAATT Xhol ORF 2 Forward GCGGATCCCATATG-TTTGATTTCGGTTTGGG BamHI-Ndel Reverse CCCGCTCGAG-GACGGCATAACGGCG Xhol ORF 2-1 Forward GCGGATCCCATATG-TTTGATTTCGGTTTGGG BamHI-Ndel Reverse CCCGCTCGAG-TGATTTACGGACGCGCA Xhol ORF 4 Forward GCGGATCCCATATG-TGCGGAGGTCAAAAAGAC BamHI-Ndel Reverse CCCGCTCGAG-TTTGGCTGCGCCTTC Xhol ORF 5 Forward GGAATTCCATATGGCCATGG-TGGAAGGCGCACAACC Ndel-Ncol Forward CGGGATCC-ATGGAAGGCGCACAAC BamHl Reverse CCCGCTCGAG-GACTGTGCAAAAACGG Xhol ORF 6 Forward CGCGGATCCCATATG-ACCCGTCAATCTCTGCA BamHI-Ndel Reverse CCCGCTCGAG-TGCGCCGAACACTTTC Xhol ORF 7 Forward CGCGGATCCGCTAGC-GCGCTGCTTTTTGTTCC BamHI-Nhel Reverse CCCGCTCGAG-TTTCAAAATATATTTGCGGA Xhol ORF 8 Forward GCGGATCCCATATG-GCTCAACTGCTTCGTAC BamHI-Ndel Reverse CCCGCTCGAG-AGCAGGCTTTGGCGC Xhol ORF 9 Forward CGCGGATCCCATATG-CCGAAGGAAGTCGGAAA BamHI-Ndel Reverse CCCGCTCGAG-TTTCCGAGGTTTTCGGG Xhol ORF IO Forward GCGGATCCCATATG-GACACAAAAGAAATCCTC BamHI-Ndel Reverse CCCGCTCGAG- TAATGGGAAACCTTGTTTT Xhol ORF ll Forward GCGGATCCCATATG-GCGGTCAACCTCTACG BamHI-Ndel Reverse CCCGCTCGAG-GGAAACGACTTCGCC Xhol ORF 13 Forward CGCGGATCCCATATG-GCTCTGCTTTCCGCGC BamHI-Ndel Reverse CCCGCTCGAG-AGGGTGTGTGATAATAAG Xhol ORF 15 Forward GGAATTCCATATGGCCATGG-GCGGGACACTGACAG Ndel-Ncol Forward CGGGATCC-TGCGGGACACTGACAGG BamHl Reverse CCCGCTCGAG-AGGTTGGCCTTGTCTATG Xhol ORF 17 Forward GGAATTCCATATGGCCATGG -TTGCCGGCCTGTTCG Ndel-Ncol Forward CGGGATCC-ATTGCCGGCCTGTTCG BamHl Reverse CCCGCTCGAG-AAGCAGGTTGTACAGC Xhol ORF 18 Forward GCGGATCCCATATG-ATTTTGCTGCATTTGGAT BamHI-Ndel Reverse CCCGCTCGAG-TCTTCCAATTTCTGAAAGC Xhol ORF 19 Forward GGAATTCCATATGGCCATGG -TCGCCAGTGTTTTTACC Ndel-Ncol Forward CGGGATCC-TTCGCCAGTGTTTTTACCG BamHl Reverse CCCGCTCGAG-GGTGTTTTTGAAGCTGCC Xhol ORF 20 Forward GGAATTCCATATGGCCATGG -TCGGCGCGGGTATG Ndel-Ncol Forward CGGGATCC-TTCGGCGCGGGTATG BamHl Reverse CCCGCTCGAG-CGGCGAGCGAGAGCA Xhol ORF 22 Forward GGAATTCCATATGGCCATGG-TGATTAAAATCAAAAAAGGTCT Ndel-Ncol Forward CGGGATCC-ATGATTAAAATCAAAAAAGGTCTAAACC BamHl Reverse CCCGCTCGAG-ATTATGATAGCGGCCC Xhol ORF 23 Forward CGCGGATCCCATATG-GATGTTTCTGTTTCAGAC BamHI-Ndel Reverse CCCGCTCGAG-TTTAAACCGATAGGTAAACG Xhol ORF 24 Forward GGAATTCCATATGGCCATGG -TGATGCCGGAAATGGTG Ndel-Ncol Forward CGGGATCC-ATGATGCCGGAAATGGTG BamHl Reverse CCCGCTCGAG-TGTCAGCGTGGCGCA Xhol ORF 25 Forward GCGGATCCCATATG-TATCGCAAACTGATTGC BamHI-Ndel Reverse CCCGCTCGAG-ATCGATGGAATAGCCG Xhol ORF 26 Forward GCGGATCCCATATG -CAGCTGATCGACTATTC BamHI-Ndel Reverse CCCGCTCGAG-GACATCGGCGCGTTTT Xhol ORF 27 Forward GGAATTCCATATGGCCATGG-AGACCTATTCTGTTTA Ndel-Ncol Forward CGGGATCC- CAGACCTATTCTGTTTATTTTAATC BamHl Reverse CCCGCTCGAG-GGGTTCGATTAAATAACCAT Xhol ORF 28 Forward GGAATTCCATATGGCCATGG-ACGGCTGTACGTTGATGT Ndel-Ncol Forward CGGGATCC-AACGGCTGTACGTTGATG BamHl Reverse CCCGCTCGAG-TTTGTCAGAGGAATTCGCG Xhol ORF 29 Forward GCGGATCCCATATG -AACGGTTTGGATGCCCG BamHI-Ndel Forward CGCGGATCCGCTAGC-AACGGTTTGGATGCCCG BamHI-Nhel Reverse CCCGCTCGAG-TTTGTCTAAGTTCCTGATATG Xhol ORF 32 Forward CGCGGATCCCATATG-AATACTCCTCCTTTTG BamHI-Ndel Reverse CCCGCTCGAG-GCGTATTTTTTGATGCTTTG Xhol ORF 33 Forward GCGGATCCCATATG -ATTGATAGGGATCGTATG BamHI-Ndel Reverse CCCGCTCGAG-TTGATCTTTCAAACGGCC Xhol ORF 35 Forward GCGGATCCCATATG-TTCAGAGCTCAGCTT BamHI-Ndel Forward CGCGGATCCGCTAGC-TTCAGAGCTCAGCTT BamHI-Nhel Reverse CCCGCTCGAG-AAACAGCCATTTGAGCGA Xhol ORF 37 Forward GCGGATCCCATATG-GATGACGTATCGGATTTT BamHI-Ndel Reverse CCCGCTCGAG-ATAGCCCGCTTTCAGG Xhol ORF 58 Forward CGCGGATCCGCTAGC-TCCGAACGCGAGTGGAT BamHI-Nhel Reverse CCCGCTCGAG-AGCATTGTCCAAGGGGAC Xhol ORF 65 Forward GGAATTCCATATGGCCATGG -TGCTGTATCTGAATCAAG Ndel-Ncol Forward CGGGATCC-TTGCTGTATCTGAATCAAGG BamHl Reverse CCCGCTCGAG-CCGCATCGGCAGACA Xhol ORF 66 Forward GCGGATCCCATATG-TACGCATTTACCGCCG BamHI-Ndel Reverse CCCGCTCGAG-TGGATTTTGCAGAGATGG Xhol ORF 72 Forward CGCGGATCCCATATG- AATGCAGTAAAAATATCTGA BamHI-Ndel Reverse CCCGCTCGAG-GCCTGAGACCTTTGCAA Xhol ORF 73 Forward GCGGATCCCATATG-AGATTTTTCGGTATCGG BamHI-Ndel Reverse CCCGCTCGAG-TTCATCTTTTTCATGTTCG Xhol ORF 75 Forward GCGGATCCCATATG- TCTGTCTTTCAAACGGC BamHI-Ndel Reverse CCCGCTCGAG-TTTGTTTTTGCAAGACAG Xhol ORF 76 Forward GATCAGCTAGCCATATG-AAACAGAAAAAAACCGC Nhel-Ndel Reverse CGGGATCC-TTACGGTTTGACACCGTT BamHl ORF 79 Forward CGCGGATCCCATATG-GTTTCCGCCGCCG BamHI-Ndel Reverse CCCGCTCGAG-GTGCTGATGCGCTTCG Xhol ORF 83 Forward GCGGATCCCATATG-AAAACCCTGCTGCTGC BamHI-Ndel Reverse CCCGCTCGAG-GCCGCCTTTGCGGC Xhol ORF 84 Forw.ard GCGGATCCCATATG-GCAGAGATCTGTTTG BamHI-Ndel Reverse CCCGCTCGAG-GTTTGCCGATCCGACCA Xhol ORF 85 Forward CGCGGATCCCATATG- GCGGTTTGGGGCGGA BamHI-Ndel Reverse CCCGCTCGAG-TCGGCGCGGCGGGC Xhol ORF 89 Forward GGAATTCCATATGGCCATGG-CCATACCTTCTTATCA Ndel-Ncol Forward CGGGATCC-GCCATACCTTCTTATCAGAG BamHl Reverse CCCGCTCGAG-TTTTTTGCGATTAGAAAAAGC Xhol ORF 97 Forward GCGGATCCCATATG-CATCCTGCCAGCGAAC BamHI-Ndel Reverse CCCGCTCGAG-TTCGCCTACGGTTTTTTG Xhol ORF 98 Forward GCGGATCCCATATG-ACGGTAACTGCGG BamHI-Ndel Reverse CCCGCTCGAG-TTGTTGTTCGGGCAAATC Xhol ORF IOO Forward GCGGATCCCATATG-TCGGGCATTTACACCG BamHI-Ndel Reverse CCCGCTCGAG-ACGGGTTTCGGCGGAA Xhol ORF lOl Forward GCGGATCCCATATG-ATTTATCAAAGAAACCTC BamHI-Ndel Reverse CCCGCTCGAG-TTTTCCGCCTTTCAATGT Xhol ORF 102 Forward GCGGATCCCATATG-GCAGGGCTGTTTTACC BamHI-Ndel Reverse CCCGCTCGAG-AAACGGTTTGAACACGAC Xhol ORF 103 Forward GCGGATCCCATATG-AACCACGACATCAC BamHI-Ndel Reverse CCCGCTCGAG-CAGCCACAGGACGGC Xhol ORF 104 Forward GCGGATCCCATATG-ACGTGGGGAACGC BamHI-Ndel Reverse CCCGCTCGAG-GCGGCGTTTGAACGGC Xhol ORF 105 Forward GCGGATCCCATATG-ACCAAATTTCAAACCCCTC BamHI-Ndel Reverse CCCGCTCGAG-TAAACGAATGCCGTCCAG Xhol ORF 106 Forward GCGGATCCCATATG-AGGATAACCGACGGCG BamHI-Ndel Reverse CCCGCTCGAG-TTTGTTCCCGATGATGTT Xhol ORF 109 Forward GCGGATCCCATATG-GAAGATTTATATATAATACTCG BamHI-Ndel Reverse CCCGCTCGAG-ATCAGCTTCGAACCGAAG Xhol ORFl 10 Forward AAAGAATTC-ATGAGTAAATCCCGTAGATCTCCC EcoRI Reverse AAACTGCAG-GGAAAACCACATCCGCACTCTGCC PstI ORFl 11 Forward AAAGAATTC-GCACCGCAAAAGGCAAAAACCGCA EcoRI Reverse AAACTGCAG-TCTGCGCGTTTTCGGGCAGGGTGG PstI ORFl 13 Forward AAAGAATTC-ATGAACAAAACCCTCTATCGTGTGATTTTCAACCG EcoRI Reverse AAACTGCAG-TTACGAATGCCTGCTTGCTCGACCGTACTG PstI ORF115 Forward AAAGAATTC-TTGCTTGTGCAAACAGAAAAAGACGG EcoRI Reverse AAAAAAGTCGAC-CTATTTTTTAGGGGCITTTGCrTGTTTGAAAAGCCTGCC Sail ORF119 Forward AAAGAATTC-TACAACATGTATCAGGAAAACCAATACCG EcoRI Reverse AAACTGCAG-TTATGAAAACAGGCGCAGGGCGGTTTTGCC PstI ORF120 Forward AAAGAATTC-GCAAGGCTACCCCAATCCGCCGTG EcoRI Reverse AAACTGCAG-CGGTTTGGCTGCCTGGCCGTTGAT PstI ORF121 Forward AAAGAATTC-GCCTTGGTCTGGCTGGTTTTCGC EcoRI Reverse AAACTGCAG-TCATCCGCCACCCCACCTCGGCCATCCATC PstI ORF 140 Forward GCGGATCCCATATG-TTGCCCACAGGCAGC BamHI-Ndel Reverse CCCGCTCGAG-GACGATGGCAAACAGC Xhol ORF 141 Forward GCGGATCCCATATG-CCGTCTGAAGCAGTCT BamHI-Ndel Reverse CCCGCTCGAG-ATCTGTTGTTTTTAAAATATT Xhol ORF 142 Forward GCGGATCCCATATG-GATAATTCTGGTAGTGAAG BamHI-Ndel Reverse CCCGCTCGAG-AAACGTATAGCCTACCT Xhol ORF 143 Forward GCGGATCCCATATG-GATACCGCTTTGAACCT BamHI-Ndel Reverse CCCGCTCGAG-AATGGCTTCCGCAATATG Xhol ORF 144 Forward GCGGATCCCATATG-ACCTTTTTACAACGTTTGC BamHI-Ndel Reverse CCCGCTCGAG-AGATTGTTGTTGTTTTTTCG Xhol ORF 147 Forward GCGGATCCCATATG-TCTGTCTTTCAAACGGC BamHI-Ndel Reverse CCCGCTCGAG-TTTGTTTTTGCAAGACAG Xhol NB: - restriction sites are underlined - for ORFs 110-130, where the ORF itself carries an EcoRI site (eg. ORF122), a Sail site was used in the forward primer instead. Similarly, where the ORF carries a PstI site (eg. ORFs 115 .and 127), a Sail site was used in the reverse primer. TABLE II - Summary of cloning, expression and purification

Claims: CLAIMS 1. A protein comprising .an amino acid sequence selected from the group consisting of SEQ IDs 2, 4, 6, and 8. 2. A nucleic acid molecule which encodes a protein according to claim 1. 3. A nucleic acid molecule according to claim 2, comprising a nucleotide sequence selected from the group consisting of SEQ IDs 1, 3, 5, and 7. 4. A protein comprising an amino acid sequence selected from the group consisting of SEQ IDs 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100, 102, 104, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 126, 128, 130, 132, 134, 136, 138, 140, 142, 144, 146, 148, 150, 152, 154, 156, 158, 160, 162, 164, 166, 168, 170, 172, 174, 176, 178, 180, 182, 184, 186, 188, 190, 192, 194, 196, 198, 200, 202, 204, 206, 208, 210, 212, 214, 216, 218, 220, 222, 224, 226, 228, 230, 232, 234, 236, 238, 240, 242, 244, 246, 248, 250, 252, 254, 256, 258, 260, 262, 264, 266, 268, 270, 272, 274, 276, 278, 280, 282, 284, 286, 288, 290, 292, 294, 296, 298, 300, 302, 304, 306, 308, 310, 312, 314, 316, 318, 320, 322, 324, 326, 328, 330, 332, 334, 336, 338, 340, 342, 344, 346, 348, 350, 352, 354, 356, 358, 360, 362, 364, 366, 368, 370, 372, 374, 376, 378, 380, 382, 384, 386, 388, 390, 392, 394, 396, 398, 400, 402, 404, 406, 408, 410, 412, 414, 416, 418, 420, 422, 424, 426, 428, 430, 432, 434, 436, 438, 440, 442, 444, 446, 448, 450, 452, 454, 456, 458, 460, 462, 464, 466, 468, 470, 472, 474, 476, 478, 480, 482, 484, 486, 488, 490, 492, 494, 496, 498, 500, 502, 504, 506, 508, 510, 512, 514, 516, 518, 520, 522, 524, 526, 528, 530, 532, 534, 536, 538, 540, 542, 544, 546, 548, 550, 552, 554, 556, 558, 560, 562, 564, 566, 568, 570, 572, 574, 576, 578, 580, 582, 584, 586, 588, 590, 592, 594, 596, 598, 600, 602, 604, 606, 608, 610, 612, 614, 616, 618, 620, 622, 624, 626, 628, 630, 632, 634, 636, 638, 640, 642, 644, 646, 648, 650, 652, 654, 656, 658, 660, 662, 664, 666, 668, 670, 672, 674, 676, 678, 680, 682, 684, 686, 688, 690, 692, 694, 696, 698, 700, 702, 704, 706, 708, 710, 712, 714, 716, 718, 720, 722, 724, 726, 728, 730, 732, 734, 736, 738, 740, 742, 744, 746, 748, 750, 752, 754, 756, 758, 760, 762, 764, 766, 768, 770, 772, 774, 776, 778, 780, 782, 784, 786, 788, 790, 792, 794, 796, 798, 800, 802, 804, 806, 808, 810, 812, 814, 816, 818, 820, 822, 824, 826, 828, 830, 832, 834, 836, 838, 840, 842, 844, 846, 848, 850, 852, 854, 856, 858, 860, 862, 864, 866, 868, 870, 872, 874, 876, 878, 880, 882, 884, 886, 888, 890, & 892.. 5. A protein having 50% or greater sequence identity to a protein according to claim 4. 6. A protein comprising a fragment of an amino acid sequence selected from the group consisting of SEQ IDs 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100, 102, 104, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 126, 128, 130, 132, 134, 136, 138, 140, 142, 144, 146, 148, 150, 152, 154, 156, 158, 160, 162, 164, 166, 168, 170, 172, 174, 176, 178, 180, 182, 184, 186, 188, 190, 192, 194, 196, 198, 200, 202, 204, 206, 208, 210, 212, 214, 216, 218, 220, 222, 224, 226, 228, 230, 232, 234, 236, 238, 240, 242, 244, 246, 248, 250, 252, 254, 256, 258, 260, 262, 264, 266, 268, 270, 272, 274, 276, 278, 280, 282, 284, 286, 288, 290, 292, 294, 296, 298, 300, 302, 304, 306, 308, 310, 312, 314, 316, 318, 320, 322, 324, 326, 328, 330, 332, 334, 336, 338, 340, 342, 344, 346, 348, 350, 352, 354, 356, 358, 360, 362, 364, 366, 368, 370, 372, 374, 376, 378, 380, 382, 384, 386, 388, 390, 392, 394, 396, 398, 400, 402, 404, 406, 408, 410, 412, 414, 416, 418, 420, 422, 424, 426, 428, 430, 432, 434, 436, 438, 440, 442, 444, 446, 448, 450, 452, 454, 456, 458, 460, 462, 464, 466, 468, 470, 472, 474, 476, 478, 480, 482, 484, 486, 488, 490, 492, 494, 496, 498, 500, 502, 504, 506, 508, 510, 512, 514, 516, 518, 520, 522, 524, 526, 528, 530, 532, 534, 536, 538, 540, 542, 544, 546, 548, 550, 552, 554, 556, 558, 560, 562, 564, 566, 568, 570, 572, 574, 576, 578, 580, 582, 584, 586, 588, 590, 592, 594, 596, 598, 600, 602, 604, 606, 608, 610, 612, 614, 616, 618, 620, 622, 624, 626, 628, 630, 632, 634, 636, 638, 640, 642, 644, 646, 648, 650, 652, 654, 656, 658, 660, 662, 664, 666, 668, 670, 672, 674, 676, 678, 680, 682, 684, 686, 688, 690, 692, 694, 696, 698, 700, 702, 704, 706, 708, 710, 712, 714, 716, 718, 720, 722, 724, 726, 728, 730, 732, 734, 736, 738, 740, 742, 744, 746, 748, 750, 752, 754, 756, 758, 760, 762, 764, 766, 768, 770, 772, 774, 776, 778, 780, 782, 784, 786, 788, 790, 792, 794, 796, 798, 800, 802, 804, 806, 808, 810, 812, 814, 816, 818, 820, 822, 824, 826, 828, 830, 832, 834, 836, 838, 840, 842, 844, 846, 848, 850, 852, 854, 856, 858, 860, 862, 864, 866, 868, 870, 872, 874, 876, 878, 880, 882, 884, 886, 888, 890, & 892.. 7. An antibody which binds to a protein according to any one of claims 4 to 6. 8. A nucleic acid molecule which encodes a protein according to any one of claims 4 to 6. 9. A nucleic acid molecule according to claim 8, comprising a nucleotide sequence selected from the group consisting of SEQ EDs 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101, 103, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123, 125, 127, 129, 131, 133, 135, 137, 139, 141, 143, 145, 147, 149, 151, 153, 155, 157, 159, 161, 163, 165, 167, 169, 171, 173, 175, 177, 179, 181, 183, 185, 187, 189, 191, 193, 195, 197, 199, 201, 203, 205, 207, 209, 211, 213, 215, 217, 219, 221, 223, 225, 227, 229, 231, 233, 235, 237, 239, 241, 243, 245, 247, 249, 251, 253, 255, 257, 259, 261, 263, 265, 267, 269, 271, 273, 275, 277, 279, 281, 283, 285, 287, 289, 291 293, 295, 297, 299, 301, 303, 305, 307,309,311 313, 315, 317, 319, 321 323, 325, 327, 329, 331 333, 335, 337, 339, 341, 343, 345, 347,349,351 353, 355, 357, 359, 361 363, 365, 367, 369, 371 373, 375, 377, 379, 381,383,385, 387, 389, 391 393, 395, 397, 399, 401 403, 405, 407, 409, 411 413,415, 417,419,421, 423, 425, 427,429,431 433, 435, 437, 439, 441 443, 445, 447, 449, 451 453, 455, 457, 459, 461, 463, 465, 467, 469, 471 473, 475, 477, 479, 481 483, 485, 487, 489, 491 493, 495, 497, 499, 501, 503, 505, 507,509,511 513, 515, 517, 519, 521 523, 525, 527, 529, 531 533, 535, 537, 539, 541, 543, 545, 547,549,551 553, 555, 557, 559, 561 563, 565, 567, 569, 571 573, 575, 577, 579, 581, 583, 585, 587, 589, 591 593, 595, 597, 599, 601 603, 605, 607, 609, 611 613,615, 617, 619, 621, 623, 625, 627,629,631 633, 635, 637, 39, 641 643, 645, 647, 649, 651 653, 655, 657, 659, 661, 663, 665, 667, 669, 671 673, 675, 677, 679, 681 683, 685, 687, 689, 691 693, 695, 697, 699, 701, 703, 705, 707,709,711 713, 715, 717, 719, 721 723, 725, 727, 729, 731 733, 735, 737, 739, 741, 743, 745, 747,749,751 753, 755, 757, 759, 761 763, 765, 767, 769, 771 773, 775, 777, 779, 781, 783, 785, 787, 789, 791 793, 795, 797, 799, 801 803, 805, 807, 809, 811 813,815, 817,819,821, 823, 825, 827, 829, 831 833, 835, 837, 839, 841 843, 845, 847, 849, 851 853, 855, 857, 859, 861, 863, 865, 867, 869, 871 873, 875, 877, 879, 881 883, 885, 887, 889, & 891. 10. A nucleic acid molecule comprising a fragment of a nucleotide sequence selected from the group consisting of SEQ LDs 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101, 103, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123, 125, 127, 129, 131, 133, 135, 137, 139, 141, 143, 145, 147, 149, 151, 153, 155, 157, 159, 161, 163, 165, 167, 169, 171, 173, 175, 177, 179, 181, 183, 185, 187, 189, 191, 193, 195, 197, 199, 201, 203, 205, 207, 209, 211, 213, 215, 217, 219, 221, 223, 225, 227, 229, 231, 233, 235, 237, 239, 241, 243, 245, 247, 249, 251, 253, 255, 257, 259, 261, 263, 265, 267, 269, 271, 273, 275, 277, 279, 281, 283, 285, 287, 289, 291, 293, 295, 297, 299, 301, 303, 305, 307, 309, 311, 313, 315, 317, 319, 321, 323, 325, 327, 329, 331, 333, 335, 337, 339, 341, 343, 345, 347, 349, 351, 353, 355, 357, 359, 361, 363, 365, 367, 369, 371, 373, 375, 377, 379, 381, 383, 385, 387, 389, 391, 393, 395, 397, 399, 401, 403, 405, 407, 409, 411, 413, 415, 417, 419, 421, 423, 425, 427, 429, 431, 433, 435, 437, 439, 441, 443, 445, 447, 449, 451, 453, 455, 457, 459, 461, 463, 465, 467, 469, 471, 473, 475, 477, 479, 481, 483, 485, 487, 489, 491, 493, 495, 497, 499, 501, 503, 505, 507, 509, 511, 513, 515, 517, 519, 521, 523, 525, 527, 529, 531, 533, 535, 537, 539, 541, 543, 545, 547, 549, 551, 553, 555, 557, 559, 561, 563, 565, 567, 569, 571, 573, 575, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, 629, 631, 633, 635, 637, 639, 641, 643, 645, 647, 649, 651, 653, 655, 657, 659, 661, 663, 665, 667, 669, 671, 673, 675, 677, 679, 681, 683, 685, 687, 689, 691, 693, 695, 697, 699, 701, 703, 705, 707, 709, 711, 713, 715, 717, 719, 721, 723, 725, 727, 729, 731, 733, 735, 737, 739, 741, 743, 745, 747, 749, 751, 753, 755, 757, 759, 761, 763, 765, 767, 769, 771, 773, 775, 777, 779, 781, 783, 785, 787, 789, 791, 793, 795, 797, 799, 801, 803, 805, 807, 809, 811, 813, 815, 817, 819, 821, 823, 825, 827, 829, 831, 833, 835, 837, 839, 841, 843, 845, 847, 849, 851, 853, 855, 857, 859, 861, 863, 865, 867, 869, 871, 873, 875, 877, 879, 881, 883, 885, 887, 889, & 891.. 11. A nucleic acid molecule comprising a nucleotide sequence complementary to a nucleic acid molecule according to any one of claims 8 to 10. 12. A nucleic acid molecule comprising a nucleotide sequences having 50% or greater sequence identity to a nucleic acid molecule according to any one of claims 8-11. 13. A nucleic acid molecule which can hybridise to a nucleic acid molecule according to any one of claims 8-12 under high stringency conditions. 14. A composition comprising a protein, a nucleic acid molecule, or an .antibody according to any preceding claim. 15. A composition according to claim 14 being a vaccine composition or a diagnostic composition. 16. A composition according to claim 14 or claim 15 for use as a pharmaceutical. 17. The use of a composition according to claim 14 in the manufacture of a medicament for the treatment or prevention of infection due to Neisserial bacteria.

Inventors: GRANDI GUIDO, MASIGNANI VEGA, PIZZA MARIAGRAZIA, RAPPUOLI RINO, SCARLATO VINCENZO, GRANDI, GUIDO, MASIGNANI, VEGA, PIZZA, MARIAGRAZIA, RAPPUOLI, RINO, SCARLATO, VINCENZO
Applicants: CHIRON SPA, GRANDI GUIDO, MASIGNANI VEGA, PIZZA MARIAGRAZIA, RAPPUOLI RINO, SCARLATO VINCENZO, CHIRON S.P.A., GRANDI, GUIDO, MASIGNANI, VEGA, PIZZA, MARIAGRAZIA, RAPPUOLI, RINO, SCARLATO, VINCENZO
Application year: 1998
Main classification: C12N15
IPC codes: A61K38, A61K39, A61K48, A61P31, C07K14, C07K16, C12N15, C12Q1