Abstract:
Nucleic acids (SEQ ID NOs: 1-3) encoding a  Plasmodium falciparum  liver stage antigen, the LSA-3 immunogenic polypeptide, recombinant vectors containing the nucleic acids, and methods of making the polypeptide using the nucleic acids and vectors are disclosed.

Description:
BACKGROUND OF THE INVENTION 
     The parasites responsible for malaria in man display different morphologies in the human host and express different antigens depending on their location in the body. The morphological and antigenic differences of these parasites during their life cycles in man enable different stages of development in the liver and in the blood to be defined: the sporozoite, the infectious form injected by the vector mosquito, transforms rapidly into a schizont in the host&#39;s hepatocytes and thereafter infects the erythrocytes. The intrahepatic localization of  P.falciparum  manifests itself in the expression of a group of antigens specific to this stage of development and which are highly immunogenic under the natural conditions of exposure to the disease. This clinically silent phase is at present the only one against which a very strong, sterilizing immunity can be induced experimentally in man, by injecting irradiated sporozoites capable of entering the hepatocyte and of developing therein but without being able to lead on to the blood stage of the disease. Accordingly, the inventors have concentrated the bulk of their efforts on these two pre-erythrocytic stages. However, these stages are also the most intricate ones to study, and hence the least understood, since it is difficult or even impossible to obtain biological material, the only in vitro study model affords a very low yield and the best animal model remains the chimpanzee, the use of which is limited and expensive. 
     In order to gain access to the antigens of the pre-erythrocytic stages, the inventors used sera of individuals who had resided for 25 years in a region where the disease is endemic but who were on permanent prophylaxis with chloroquine. These individuals were regularly subjected to infected mosquito bites but did not develop any complete blood infection. Their serum hence contained antibodies directed essentially against the pre-erythrocytic stages, which was verified by immunofluorescence (IF) and western blotting on the 3 stages of the parasite. 
     The use of these sera for screening a library of genomic DNA of the parasitic clone of  P.falciparum,  the library being constructed in expression vectors in a phage lambda gt11 (V. Rosario, Science 212, 1981, pp. 1037-1038; and Thaithong et al., Transactions of Royal Society of Tropical Medicine and Hygiene, 1984, 78:242-245), led to the demonstration of polypeptides of the pre-erythrocytic stage, in particular the SALSA (sporozoite liver stage antigen) polypeptides described in EP A-0,407,230 and LSA-1 (liver stage antigen) described in WO 92/13884. The present invention relates to new polypeptide molecules specific to the pre-erythrocytic stage, and to their use as active principle of antimalarial vaccine or in methods of diagnosis of the disease. 
     SUMMARY OF THE INVENTION 
     The invention is the outcome of the demonstration by the inventors of the special properties of a particular antigen referred to as LSA-3 and of its fragments, which are seen to be candidates with a strong potential for producing an antimalarial vaccine, for the following reasons: 
     a) when a fraction of LSA-3 was used in combination with another antigen of the same stage of development of the parasite, such as LSA-1, to immunize chimpanzees, the animal responding to both molecules or only to LSA-3 displays the feature of not having parasites in the blood, of having a substantial decrease of the parasites in the liver and of manifesting a substantial recruitment of mononuclear cells indicating a response in terms of cellular immunity; 
     b) in regions where the disease is endemic, a very clear correlation is observed between the protection of individuals against natural infection by sporozoites and their responses in terms of antibodies against LSA-3; 
     c) in eight human volunteers immunized by injection of irradiated sporozoites, antibodies directed against LSA-3 are found in each of the four individuals resisting sporozoite infection and in none of the other four volunteers who developed a blood infection; 
     d) antibodies obtained against the peptide DG729 in WO 92/13884, already described, give a cross-reaction with the sporozoite and liver stages of the murine parasite  P.yoelii,  which permits a significant exploitation of the mouse model. In vitro, the human antibodies immunopurified on DG729 are capable, even at very low concentrations, of blocking the entry of  P.yoelii  sporozoites into mouse hepatocytes. In vivo, mice immunized with DG729 are fully or partially protected against infection by  P.yoelii  sporozoites; 
     e) lastly, some epitopes, in particular in the non-repeat portions of the molecule, stimulate the secretion of interferon-γ by monocytes, this mediator enabling the intrahepatic development of the parasite to be inhibited (S. Mellouk et al., The Jour. Of Immun. 139, 4192-4195, 1987); 
     f) the sequence of the region of LSA-3 corresponding to a (lipo)peptide NR2 was analysed in 27 samples: 4 laboratory strains (NF54, K1, Palo Alto, T9/96), 3 Madagascan isolates, 3 Burmese isolates, 5 Brazilian isolates, 7 isolates from the Ivory Coast and 5 Thai isolates. No mutation was observed on the 300 base pairs analysed, that is to say 100% conservation in this immunologically important region containing one or more B, Th and CTL epitopes; 
     g) information about the structure of the antigen, and in particular of a peptide RE, and more especially about the central repeat region from which the peptide RE was designed and which contains one or more major B epitopes, was obtained from the hydrophobic cluster plot of the sequence available in the clone T9/96 (630 amino acids) (Gaboriot et al., (1987): Hydrophobic cluster analysis: an efficient new way to compare and analyse amino acid sequences, FEES Letters, 224: 149-155); this method predicts a very strong propensity for α-helical organization. The repeat region displays remarkable regularity in the spacing of the valine and isoleucine residues, alternating with acid or proline residues. The arrangement of the hydrophobic groups at the surface of this helix is reminiscent of a hydrophobic border gradually shifting from one face of the helix to the other according to a constant general orientation along the molecule, and probably related to a coiled-coil structure or packaging as seen in FIG. 4 b  which depicts the HCP (hydrophobic cluster plot) of the peptide sequence of the clone DG729; 
     h) after demonstrating that there was a very wide range of immune responses to the LSA-3 antigen, we analysed the capacity of the responder cells to localize around the parasites in the liver. In mice immunized with the recombinant antigens, intraportal injection of each of the peptides absorbed on 10 μm polystyrene beads enables an afflux of lymphocytes around the antigen (mimicking the parasite) to be visualized after 48 hours, followed on the 5th day by a substantial recruitment of cells belonging to the macrophage line. 
     All these properties, some of which will be demonstrated in detail in the experiments described later, show that the LSA-3 antigen displays both good antigenicity and good immunogenicity. 
     The inventors were able to confirm and define the specificity of the stages of expression of the molecule; in the sporozoites, this expression was confirmed by the surface immunofluorescence of several strains and isolates. In western blot (or immunoblot) analysis, the LSA-3 molecule appears as a protein of molecular weight 200,000 daltons. While the messenger RNAs of sporozoites could not be obtained in sufficient amounts for a northern blot analysis, reverse PCR experiments confirmed the expression of LSA-3 at this stage. In infected hepatocytes, LSA-3 is observed in the parasitophorous vacuole of the parasite by immunofluorescence using antibodies against the repeat and non-repeat regions of the protein, as well as by electron microscopy. 
     A fragment of LSA-3 designated 729S, as well as three peptides designated NRI and NRII included in the non-repeat portion and 729R included in the repeat portion, have been described in Application WO 92/13884. Nevertheless, this document does not mention the special properties mentioned above, or other fragments of LSA-3 which could be either longer or shorter, included or combined with these fragments, which might display especially advantageous properties for use in vaccines. 
     The subject of the invention is polypeptide molecules containing at least ten consecutive amino acids of the amino acid sequence shown in FIG.  2  and designated SEQ ID No. 8, and representing LSA-3, the following polypeptides being excluded SEQ ID NO: 10-15: 
     RDELFNELLNSVDVNGEVKENILEESQVNDDIFNSLVKSVQQEQQHNVEE VEESVEENDEESVEENVEENVENNDDGSVASSVEESIASSVDESIDSSIE-ENVAPTVEEIVAPTVEEIVAPSVVEKCAPSVEESVAPSVEESVAEMLKER (729S) 
     RDELFNELLNSVDVNGEVKENILEESQVNDDIFNSLVKSVQQEQQHN DELFNELLNSVDVNGEVKENILEESQ, (NRI) 
     LEESQVNDDIFSNSLVKSVQQEQQHNV, (NRII) 
     VESVAPSVEESVAPSVEESVAENVEESV. (729RE) 
     Other molecules according to the invention contain at least 20 consecutive amino acids or at least 50. 
     This set of polypeptides and the LSA-3 molecule are, throughout hereinafter, “polypeptides of the invention”. 
     The experimental results and the comparisons of non-repeat sequences between different  P.falciparum  isolates indicate the existence of at least 70% homology between equivalent antigens of the liver stage of the parasite. Thus any peptide molecule displaying at least 70% homology with any one of the molecules defined above forms part of the invention, as do those displaying at least 70% homology with the following sequence (SEQ ID NO: 16): 
     Leu Leu Ser Asn Ile Glu Glu Pro Lys Glu Asn Ile Ile Asp Asn Leu Leu Asn Asn Ile (CT1) 
     lying between amino acids 140 and 159 of K1 or 23 and 42 of T9/96. 
     Likewise forming part of the invention are the polypeptide molecules displaying at least 70% homology with the sequence depicted in FIG. 3, which depicts a portion of LSA-3 in T9/96: the DNA of this  P.falciparum  isolate was digested with restriction enzymes, then cloned into lambda gt11 and thus enabled the gene library of this isolate, already described above, to be constituted. 
     Conjugates consisting of a polypeptide originating from LSA-3 linked covalently via a lysine bridge to saturated or unsaturated lipid residues also form part of the invention, more especially when the lipid residue is a palmitoyl or a palmityl or an oleyl. C 16  or C 18  residues were thus coupled via a lysine bridge to the peptides NRI, NRII, 729RE and CT1 already depicted above. The method of synthesis used for these conjugates is described in Bourgault, Journal of Immunology, 149, 3416 (1992) and Rouaix, Vaccine, 12, 1209 (1994). 
     The invention also covers immunogenic compositions containing at least one polypeptide molecule or one conjugate described above, as well as the vaccines containing these immunogenic compositions. Other immunogenic epitopes, in particular LSA-1, SALSA and STARP, have already been described in EP A-0407230 and in WO 92/13884. The vaccine compositions according to the invention can advantageously contain a mixture of immunogenic peptides originating from LSA-3 and of the peptides or antigens originating from LSA-1, SALSA or STARP; a more especially advantageous mixture could be the one consisting, on the one hand of NRI, NRII or whole LSA-3, these being coupled or otherwise to a lipid residue, and on the other hand the peptides SALSA-1, SALSA-2 or the SALSA antigen coupled or otherwise to a lipid residue. 
     All polypeptide molecules corresponding to the above definition and displaying at least 70% homology with the polypeptides LSA-3, CT1, NRI, NRII or 729RE may be combined in homologous or heterologous fashion with other peptide sequences or sequences originating from another antigen of the different stages of  P.falciparum.    
     70% Homology of sequences should be clearly understood to refer to a sequence homology with respect to any one of the isolates whose sequence is known or capable of being known, and not an overall homology between the collective isolates. In effect, the central repeat region of LSA-3 (block 2 of FIG. 4) displays a variable number of repeat sequences responsible for a variability from one isolate to another, as seen, moreover, in the diagram of FIG. 4, in which the difference in length between the repeat portions of block 2 of the isolates T9/96 and K1 is blatant although the tetrapeptides which constitute this repeat region (VEES, VEEN, VEEI, VAPS, VAPT SEQ ID NO: 17-20 and the like) are very well conserved. In contrast, the repeat sequences of block 1 are fully conserved between the two isolates. Thus, bearing in mind the intrinsic variability of this block 2 from one isolate to another, the definition 70% homology applies to the LSA-3 antigen of the different isolates excluding the repeat sequences of block 2. 
     The invention also covers the polyclonal or monoclonal antibodies which specifically recognize the polypeptide molecules of the invention. 
     These molecules of the invention may be used for carrying out diagnostic methods and producing kits enabling the existence of  P.falciparum  infection to be detected; this method can be either an assay of circulating specific antibodies, by carrying out standard serological methods by bringing one of the above antigens into contact with a biological fluid of the individual in question, or methods of assay of antigens using polyclonal or monoclonal antibodies obtained by standard methods for obtaining such antibodies with the corresponding antigens. In the diagnostic outfits or kits of the invention, the reagents enabling the antigen/antibody complexes produced to be detected, which can also carry a label or be capable of being recognized in their turn by a labelled reagent, are present. Depending on whether it is desired to carry out an antigen test or a serological test, the kit comprises either the antibodies or the antigens of the invention. 
     The invention also covers all the nucleotide sequences coding for a polypeptide of the invention, as well as any recombinant nucleic acid containing at least one nucleotide sequence of the invention, inserted into a nucleic acid which is heterologous with respect to the said nucleotide sequence. 
     The nucleic acid sequences coding for LSA-3 or its immunogenic fragments and corresponding to one of the following definitions form part of the invention: 
     (a) the linked succession of nucleotides as depicted in SEQ ID No. 1 of FIG. 1, or 
     (b) the linked succession of nucleotides depicted in SEQ ID No. 2 of FIG. 2, 
     (c) a linked succession displaying at least 70% homology with that of FIG. 1 or of FIG. 2, or 
     (d) a linked succession of nucleotides which are complementary to those presented in (a), (b) or (c). 
     The expression “coding for LSA-3” is understood to refer both to the gene depicted in SEQ ID No. 1 of FIG.  1  and the cDNA depicted in SEQ ID No. 2 of FIG.  2 . 
     The invention relates more especially to a recombinant nucleic acid in which the nucleotide sequence of the invention is preceded by a promoter (in particular an inducible promoter), under the control of which the transcription of the said sequence is capable of being performed, and, where appropriate, followed by a sequence coding for transcription termination signals. 
     The invention also covers the coding sequence originating from the clone T9/96 depicted in FIG. 3 by SEQ ID No. 3. 
     In this sequence, the fragment CT1 lies in [sic] nucleotide [sic] 67 and 126, the fragment 679 how [sic] at nucleotide 206 and the fragment 729RE lies between nucleotides 547 and 630. 
     Lastly, the invention covers any recombinant vector used especially for the cloning of a nucleotide sequence of the invention, and/or for the expression of the polypeptide encoded by this sequence, and characterized in that it contains a recombinant nucleic acid as defined above in one of its sites which is not essential for its replication. 
     As an example of an abovementioned vector, plasmids, cosmids, phages or viruses may be mentioned. 
     As such, the invention relates more especially to the plasmid pK 1.2. deposited at the Collection Nationale De Cultures De Microorganismes (CNCM), Paris, France, on May 10, 1995 under the Accession Number No. I-1573. 
     The subject of the invention is also a method for preparing a polypeptide of the invention, by transformation of a cell host using a recombinant vector of the abovementioned type, followed by the culturing of the cell host thus transformed and the recovery of the polypeptide in the culture medium. 
     Thus, the invention relates to any cell host transformed by a recombinant vector as defined above, and comprising the regulatory elements permitting the expression of the nucleotide sequence coding for a polypeptide according to the invention. 
     The invention likewise covers DNA (or RNA) primers which can be used in the context of the synthesis of nucleotide and/or polypeptide sequences of the invention, by the PCR (polymerase chain reaction) technique or any other method known at the present time for amplifying nucleic acids, such as LCR, CPR, ERA, SPA, NASBA, and the like. 
     The invention relates to any DNA or RNA primer, characterized in that it consists of approximately 10 to 25 nucleotides which are identical or complementary to the first 10 to 25 nucleotides of the nucleotide sequence coding for a peptide sequence according to the invention, or identical to the last 10 to 25 nucleotides of the said sequence. 
     Thus, the present invention also covers a method for preparing a polypeptide of the invention comprising the following steps: 
     where appropriate, the prior amplification by standard techniques of the amount of nucleotide sequences coding for the said polypeptide using two suitably chosen DNA primers, 
     the culturing, in a suitable culture medium, of a cell host previously transformed by a vector containing a nucleic acid according to the invention comprising the nucleotide sequence coding for the said polypeptide, and 
     the recovery from the abovementioned culture medium of the polypeptide produced by the said transformed cell host. 
     By way of example of DNA or RNA primers according to the invention, the following pairs of sequences may be mentioned: 
     S1: GTGATGAACTTTTTAATGAATTATTAAA (SEQ ID No. 4) 
     S2: TGTTGTTCTTGTTGAACACTTTTTACTAA (SEQ ID No. 5) 
     whose respective positions on the LSA-3/K1 gene depicts [sic] in FIG. 1 are from 695 to 722 and from 829 to 799 (reading in the reverse direction), or the pair: 
     6.1: GGTATCGAAACTGAGGAAATAAAGG (SEQ ID No. 6) 
     6.2: CATAGCAGGAACATCAACATCCAC (SEQ ID No. 7) 
     whose respective positions are 2668 to 2692 for 6.1 and 3456 to 3433 for 6.2 (reading in the reverse direction). 
     The information regarding the sequences ID No. 4, ID No. 5, ID No. 6 and ID No. 7 are detailed at the end of the description. 
     The peptides of the invention may also be prepared by the standard techniques of peptide synthesis. This synthesis may be carried out in homogeneous solution or in the solid phase. For example, use may be made of the technique of synthesis in homogeneous solution described by Houben-Weyl in the work entitled “Methoden der Organischen Chemie” (Methods in Organic Chemistry) edited by E. Wunsch, vol. 15-I and II. Thieme, Stuttgart 1974, or that described by R. D. Merrifield in the paper entitled “Solid phase peptide synthesis” (J. Am. Chem. Soc., 45, 2149-2154). 
     The invention also covers the water-soluble oligomers of the abovementioned monomeric peptides. 
     Oligomerization can cause an enhancement of the immunogenicity of the monomeric peptides according to the invention. While such numerical information cannot be regarded as limiting, it may nevertheless be mentioned that these oligomers can, for example, contain from 2 to 10 monomer units. 
     To carry out the oligomerization, use may be made of any polymerization technique commonly used in the peptide field, this polymerization being conducted until an oligomer or polymer containing the requisite number of monomer motifs for acquiring the desired immunogenicity is obtained. 
     One method of oligomerization or polymerization of the monomer consists in reacting the latter with a crosslinking agent such as glutaraldehyde. 
     Use may also be made of other oligomerization or coupling methods, for example the one employing successive couplings of monomer units via their carboxy- and amino-terminal functions in the presence of homo- or heterobifunctional coupling agents. 
     The invention also relates to the conjugates obtained by covalent coupling of the peptides according to the invention (or of the abovementioned oligomers) to physiologically acceptable and non-toxic (natural or synthetic) carrier molecules that enable, in particular, the immunogenicity to be argued [sic], via complementary reactive groups carried, respectively, by the carrier molecule and the peptide. By way of example of macromolecular carrier molecules or supports which participate in the constitution of the conjugates according to the invention, there may be mentioned natural proteins such as tetanus toxoid, ovalbumin, serum albumins, haemocyanins, tuberculin PPD (PPD: purified protein derivative), and the like. 
     By way of synthetic macromolecular supports, my [sic] may be mentioned, for example, polylysines or poly(DL-alanine)-poly(L-lysine)s. 
     By way of hydrocarbon or lipid supports, there may be mentioned saturated or unsaturated fatty acids, and preferably C 16  or C 18  acids of the oleyl [sic] or palmitoleyl [sic] type. 
     Lastly and without implied limitation, the antigens or peptides according to the invention may be coupled to traditional supports or adsorbed on such supports, in particular latex or polystyrene micro-spheres or beads, or incorporated in Ty1 particles. 
     To synthesize the conjugates according to the invention, use may be made of methods which are known per se, such as the one described by Frantz and Robertson in Infect. and Immunity, 33, 193-198 (1981), or the one described in Applied and Environmental Microbiology (October 1981), vol. 42, No. 4, 611-614 by P. E. Kauffman, using the peptide and the appropriate carrier molecule. 
     The nucleic acids of the invention may be prepared either by a chemical method or by other methods. 
     A suitable method of preparing the nucleic acids of the invention containing not more than 200 nucleotides (or 200 bp in the case of double-stranded nucleic acids) comprises the following steps: 
     DNA synthesis using the automated P-cyanoethyl-phosphoramidite method described in Bioorganic Chemistry 4; 274-325 (1986), 
     cloning of the nucleic acids thereby obtained into a suitable vector and recovery of the nucleic acid by hybridization with a suitable probe. 
     A chemical method of preparation of nucleic acids of length greater than 200 nucleotides has already been described in WO 92/13884. 
     The invention also relates to diagnostic kits which contain one or more amplification primers specific for the LSA-3 gene and which enable the presence of the gene or of the mRNA to be detected in an individual likely to be infected by  P.falciparum.    
     The invention also covers pharmaceutical or vaccine compositions in which at least one of the products according to the invention is present in combination with solid or liquid, pharmaceutically acceptable excipients suitable for the construction of oral, ocular or nasal dosage forms, or excipients suitable for the construction of dosage forms for rectal administration, or alternatively with gelatinous excipients for vaginal administration. It also relates to isotonic liquid compositions containing at least one of the conjugates according to the invention, suitable for administration to the mucosae, in particular the ocular or nasal or pulmonary mucosae. 
     Advantageously, the vaccine compositions according to the invention contain, in addition, a vehicle such as polyvinylpyrrolidone which facilitates the administration of the vaccine. In place of polyvinylpyrrolidone, it is possible to use any other type of adjuvant, in the traditional sense which was formerly given to this expression, that is to say a substance which enables a medicinal product to be absorbed more readily or which facilitates its action in the body. By way of examples of other adjuvants of this latter type, there may also be mentioned carboxymethylcellulose, aluminium hydroxides and phosphates, saponin or all other adjuvants of this type which are well known to a person skilled in the art. Lastly, they contain, if necessary, an immunological adjuvant, in particular of the muramyl peptide type. 
     The invention also relates to pharmaceutical compositions containing as active substance at least one of the polyclonal or monoclonal antibodies defined above, in combination with a pharmaceutically acceptable vehicle. 
     Lastly, the invention covers a method of immunization of an individual likely to be infected by  P.falciparum,  by injection of a peptide molecule or an oligomer as described above, alone or in combination with other types of molecules capable of protecting the said individual against subsequent infection; the polypeptide or antigenic molecule or the natural or recombinant lipopeptides are either used alone or adsorbed or coupled to latex or polystyrene micro-spheres or beads. 
     Additional features of the invention will also become apparent in the examples illustrated with the figures which follow, and show the special features of the molecules of the invention relative to other antigens of the pre-erythrocytic stage of the parasite. 
    
    
     BRIEF DESCRIPTION OF THE DRAWINGS 
     FIGS. 1A-1F depicts the genomic DNA sequence SEQ ID No. 1 of 6152 base pairs of the LSA-3 gene; it originates from the clone K1.2, which itself originates from a Thai isolate. 
     FIGS. 2A-2L depicts the cDNA sequence ID No. 2 and the polypeptide sequence of the LSA-3 antigen. The DNA sequence represents 5361 base pairs. The amino acid sequence shown in FIG. 2 is listed in the Sequence Listing as SEQ ID NO: 8. 
     FIG. 3 depicts the sequence ID No. 3 of the portion sequenced in the parasite clone T9/96 (1890 base pairs), the upper line being the nucleotide sequence and the lower line the peptide sequence. In this clone, the CT1 sequence lies between nucleotides 67 and 126, the actual fragment DG679 beginning at nucleotide 207. The fragment 729RE lies between nucleotides 547 and 629. The amino acid sequence shown in FIG. 3 is listed in the Sequence Listing as SEQ ID NO: 9. 
     FIG. 4 a  depicts diagrammatically the relative positions of the repeat and non-repeat sequences, the introns and the exons in strains K1 and T9/96, the clones 679 and 729 originating from the latter. 
     FIG. 4 b  depicts the HCP (hydrophobic cluster plot) of the peptide sequence of the clone DG729. 
     FIG. 5 depicts the amounts of immunoglobulins produced in the serum of chimpanzee Nuria before and after immunization with different LSA-3 peptides. 
     FIG. 6 shows the specific antibody titre of different species of mice immunized either with a peptide or with a corresponding lipopeptide. 
     FIG. 7 shows the inhibition of the sporozoite invasion of liver cells by hyperimmune sera obtained after immunization with different peptides [lacuna] immunopurified against whole LSA-3. 
     FIG. 8 depicts the comparison of an antigen originating from LSA-3 with two other antigens with respect to type T immunity. 
     FIGS. 9A,  9 B and  9 C depicts the induction of interferon-γ in the chimpanzees Gerda and Dirk with the peptides originating from the LSA-3 molecule. 
     FIGS. 10A and 10B depicts the results of lymphoproliferation of the PBMC of an individual protected by an injection of irradiated sporozoites against peptides originating from the LSA-1 and LSA-3 antigens. 
    
    
     DETAILED DESCRIPTION OF THE INVENTION 
     EXAMPLE 1 
     Cloning and sequencing of the LSA-3 gene 
     1) Sequencing 
     Initial screening of the gene library originating from the parasite clone T9/96 with the serum of a missionary treated continuously by prophylaxis enabled us to isolate 120 clones corresponding to molecules expressed at the sporozoite and/or liver stage of the  P.falciparum  cycle. The clone 729S was used as probe to screen a genomic library of the Thai strain K1 already mentioned above, which contains large EcoR I fragments cloned into phage lambda gt10. A 6.85-kilobase insert containing the whole gene was purified from this gene library and recloned into a pUC18 plasmid for sequencing and characterization (plasmid pk 1.2). In  P.falciparum,  the genome of which is very rich in bases A:T(80%), this approach is often rendered difficult by the rarity of restriction sites which can be used, and by the instability or even the impossibility of cloning certain fragments when they are inserted into plasmid vectors. 
     The structure of the gene is depicted in FIG.  4  and displays the following features: 
     a) a mini-exon 1 coding at its 3′ end for a hydrophobic signal peptide; 
     b) a short intron (168 base pairs) included between consensus splicing donor and acceptor sites; 
     c) a second exon of five kilobases which codes for an organized region of 1.8 kilobases, and composed of an arrangement of 7 blocks of 4 amino acids and a 3′ hydrophobic region which might correspond to an inking [sic] of the glycosylphosphatidylinositol (GPI) type. 
     A detailed investigation of the polymorphism of LSA-3 was carried out by sequencing the clone 679, which contains the bulk of the repeat sequences of the LSA-3 gene and a 1-kilobase portion of the 3′ non-repeat fraction, the sequence of this fragment being depicted in FIG. 3 between nucleotides 207 and 1890. 
     The strain K1 repeats are the following: 
     
       
         
               
             
               
               
               
               
               
               
             
               
               
               
               
               
             
               
             
               
               
             
               
             
               
               
               
               
               
               
               
               
               
               
             
               
               
               
               
               
             
               
               
               
               
               
               
             
               
             
           
               
                   
               
             
             
               
                   Block 1:  (aa223) VEEK VEES VEEN DEES VEEN VEEN VEEN 
               
               
                 DDGS VASS VEES IASS VDES IDSS IEEN (aa278) (SEQ ID NO:21) 
               
             
          
           
               
                 
                   Block 2: 
                 
                 (aa279) 
                 VAPT 
                 VEEIVAPS 
                 VVESVAPS 
                 VEESVEEN 
               
               
                 VEESVAEN 
                 VEESVAEN 
                 VEESVAEN 
                 VEESVAEN 
               
               
                 VEEIVAPT 
                 VEEIVAPT 
                 VEEIVAPS 
                 VVESVAPS 
                 VEESVEEN 
               
               
                 VEESVAEN 
                 VEESVAEN 
                 VEESVAEN 
                 VEESVAEN 
                 VEESVAEN 
               
               
                 VEEIVAPT 
                 VEEIVAPT 
                 VEEIVAPS 
                 VVESVAPS 
                 VEESVEEN 
               
               
                 VEESVAEN 
                 VEESVAEN 
                 VEESVAEN 
                 VEESVAEN 
                 VEESVAEN 
               
               
                 VEESVAEN 
                 VEESVAEN 
               
               
                 VEEIVAPT 
                 VEEIVAPT 
                 VEEIVAPS 
                 VVESVAPS 
                 VEESVEEN 
               
               
                 VEESVAEN 
                 VEESVAEN 
                 VEESVAEN 
                 VEESVAEN 
               
               
                 VEEIVAPT 
                 VEEIVAPT 
                 VEEIVAPS 
                 VVESVAPS 
                 VEESVEEN 
               
               
                 VEESVAEN 
                 VEESVAEN 
                 VEESVAEN 
               
               
                 VEEIVAPT 
                 VEEIVAPT 
                 VEEIVAPS 
                 VVESVAPS 
                 VEESVEEN 
               
               
                 VEESVAEN 
                 VEESVAEN 
                 VEESVAEN 
                 VEESVAEN 
               
               
                 VEEIVAPT 
                 VEEIVAPT 
                 VEEIVAPS 
                 VVESVAPS 
                 VEESVEEN 
               
               
                 VEESVAEN 
                 VEESVAEN 
                 VEESVAEN 
               
             
          
           
               
                 VEESVAPT 
                 VEEIVAPS 
                 VEESVAPS 
                 VEESVAEN 
                 (aa818) (SEQ ID NO:22) 
               
             
          
           
               
                 
                   Block 3: 
                 
               
               
                 (aa1537) DEDI EEDV EEDI EEDI EEDK VEDI DEDI DEDI GEDK 
               
               
                 DEVI (aa1576) (SEQ ID NO:23) 
               
             
          
           
               
                   
                 The repeats in the clone T9/96 as determined in 
               
             
          
           
               
                 Patent Application No. FR 91/01286 of 5th February 1991 
               
               
                 are the following: 
               
             
          
           
               
                 
                   Block 1: 
                 
                 VEEK 
                 VEES 
                 VEEN 
                 DEES 
                 VEEN 
                 VEEN 
                 VEEN 
                 DDGS 
                 VASS 
               
             
          
           
               
                 VEES 
                 IASS 
                 VDES 
                 IDSS 
                 IEEN (SEQ ID NO:24) 
               
             
          
           
               
                 
                   Block 2: 
                 
                 VAPT 
                 VEEIVAPT 
                 VEEIVAPS 
                 VVESVAPS 
                 VEESVAPS 
               
               
                 VEESVAEN 
                 VEESVAEN 
               
               
                 VEEIVAPS 
               
               
                 VEESVAEN 
                 VEESVAEN 
                 VEESVAEN 
                 VEESVAEN 
                 VEESVAEN 
               
               
                 VEEIVAPT 
                 VEESVAPT 
                 VEEIVAPT 
                 VEESVAPT 
                 VEEIVVPS 
                 VEESVAPS 
               
               
                 VEESVAEN 
                 VEESVAEN 
                 VEESVAEN 
                 VEESVAEN 
                 VEESVAEN 
               
               
                 VEEIVAPS 
                 VEEIVAPT 
               
             
          
           
               
                 VEESVAEN (SEQ ID NO:25) 
               
               
                   
               
             
          
         
       
     
     Exon 2 of the LSA-3 gene contains 2 repeat regions which can be split into 3 blocks as shown in FIG.  4 : 
     Block 1, coding for a linked succession of 14 tetra-peptides. This block is 100% conserved with respect to amino acids and nucleic acids between T9/96 and K1. Only the tetrapeptides VEES and VEEN are to be found in block 2. 
     Block 2 codes, in K1, for 127 tetrapeptides corresponding to the linked succession of different octapeptides which are themselves formed by combination of 2 of the 7 basic tetrapeptides or motifs (VEES, VVES, VEEN, VEEI, VAEN, VAPS, VAPT (SEQ ID NO: 26-28)). The number of repeats and the arrangement of these octapeptides vary according to the motifs and appear to be specific to the clone K1.2. In effect, in the clone T9/96, block 2 (53 tetrapeptides) also corresponds to the linked succession of octapeptides formed from the same 7 basic tetrapeptides (and an 8th motif VVPS (SEQ ID NO: 29) which does not exist in K1), but with a different number and arrangement of these repeats. 
     Block 3 consists of the linked succession of 10 degenerate tetrapeptides different from those of blocks 1 and 2. This block was sequenced only in the strain K1. However, preliminary results obtained by PCR with the clone T9/96 and several other laboratory strains indicate that there is no size polymorphism in this region. 
     The non-repeat regions of exon 2 are especially well conserved. In effect, sequence comparison between T9/96 and K1 could be done on 315 bp at the 5′ end of block 1 and on 763 bp at the 3′ end of block 2. The homology is 99.4% with respect to nucleic acids and 98.6% with respect to amino acids. 
     Comparison of the sequences of the clone 679 originating from  P.falciparum  clone T9/96, and of the corresponding sequence of LSA-3 originating from the isolate K1, shows that the gene is well conserved, the most significant differences being observed in the repeat region where the blocks of 4 amino acids are well conserved but vary in their number and organization. 
     In contrast, the non-repeat 5′ and 3′ portions apear to be especially well conserved, showing up to 100% homology in the 5′ region where B and T epitopes have already been identified. 
     DNA amplifications, in particular by PCR of different  P.falciparum  strains with 8 primer pairs distributed over the whole of the LSA-3 gene, showed that, except with the ones surrounding the repeat regions, the whole of the genome gives PCR products of similar size, suggesting that the LSA-3 antigen is well conserved. 
     Various LSA-3 probes, chosen in the repeat and non-repeat regions, were hybridized at low stringency with the DNAs of different species of Plasmodium, and did not enable any gene homologous to LSA-3 to be identified except in the chimpanzee parasite  P.reichenowi,  confirming the close kinship of this species with  P.falciparum.    
     Surprisingly, the antigen analogous to LSA-3 found in  P.yoelii,  which gives clear immunological cross-reactions at the surface of the sporozoite with antibodies against the fragment 729S, does not appear to be conserved at the level of the nucleotide sequence. Lastly, comparison of the LSA-3 sequences with the data bases did not reveal any homology with known molecules, except for the repeat region, some of the motifs of which display a strong analogy with the repeats of a Staphylococcus xylosis gene, but also with two  P.falciparum  antigens, RESA and Pf11.1, which are both expressed during the blood stage of the parasite. This homology is essentially due to the large amount of “Glu-Glu” sequences in these antigens and in the repeats of LSA-3. 
     2) Cloning 
     The insert DG729 and other regions of exon 2 of the strain K1 were cloned into a prokaryotic expression vector pGEX, a vector marketed by the company InVitrogen Corp (San Diego USA). This vector produces a fusion protein with the  Schistosoma mansoni  glutathione S-transferase (GST), and enables the recombinant proteins to be purified readily by affinity for glutathione-agarose beads. The expression peptides from these vectors are designated: 
     for the whole LSA-3 protein: REC protein, 
     or for the fragment 729S:729PGEX. 
     Attempts at cloning other fragments, in particular the fragment 1-5 3NSREP, 3NFREP, 5NR and 5SNREP, caused difficulties related either to the cloning or to the production and purification of the proteins in sufficient amounts for immunization experiments. 
     Only the fragments 729, NN and 3PC enabled corresponding recombinant polypeptides to be produced and purified in sufficient amounts for analysis of the antigenicity of the molecule. 
     EXAMPLE 2 
     Protection of immunized chimpanzees against challenge infections at low or high dose 
     2.1. A chimpanzee Dirk previously immunized with a fraction of LSA-3 in combination with another antigen of the same stage of development of the parasite, and displaying the effects described above in point a), was reimmunized a few years later with peptides and recombinant proteins corresponding to the same combination of antigens. Once again, this chimpanzee proves to be protected against a challenge infection at low dose (2×10 4  sporozoites) and then a challenge infection at high dose (5×10 6  sporozoites). As during the first challenge, a substantial reduction is observed in the number of schizonts detected in the liver after the challenge at high dose, as well as a lymphocytic-monocytic infiltrate around the few schizonts that are detectable (testifying to a local defence). 
     2.2 Partial protection of the chimpanzee Gerda: another chimpanzee was immunized only with the LSA-3 antigen (animal described in Examples 7 and 8 below), namely the lipopeptide NR2 and then recombinant proteins (GST-729, GST-NN, GST-3PC) which, the three of them collectively, cover 950 of the LSA-3 molecule and which are adsorbed on latex microspheres. This animal proves to be partially protected against a challenge infection at high dose (8×10 6  sporozoites), since it displays a very low blood parasitaemia and a 90% reduction in the number of liver schizonts relative to the control following the challenge infection. 
     2.3. Partial protection of the chimpanzee Nuria: a chimpanzee immunized with a fraction of the LSA-3 antigen alone, namely a combination of peptides, of lipopeptides and then of recombinant proteins corresponding to 95% of the LSA-3 molecule and emulsified in Montanide ISA-51 (SEPPIC, 75 Quai d&#39;Orsay, France), proves to be partially protected against a challenge infection at moderate dose (1×10 5  sporozoites). In effect, this animal displays a significant delay in the appearance of the parasites in the blood relative to 4 controls (chimpanzees immunized with the pre-erythrocytic antigens LSA-1, SALSA or STARP, and 1 unimmunized control animal), a lower maximum blood parasitaemia and a faster fall in parasitaemia (24 hours instead of 3 days), which results reflect a large reduction in the number of liver forms induced in this animal by the challenge infection and in agreement with the results obtained in Gerda. In this case, examination of the liver forms was not carried out. 
     2.4. B and T immunogenicity in the chimpanzees Demi, Karlien and Iris: three chimpanzees immunized with the peptides LSA-3-NR1 and -RE and the lipopeptides -NR2 and -CT1, as well as with peptides corresponding, for each animal, to another pre-erythrocytic antigen (LSA-1, SALSA or STARP), display, all 3 of them: 
     high humoral responses against the B epitopes present on the peptides NR1, NR2 and RE. The antibodies recognize not only the peptides and the recombinants but are also strongly positive on the native molecules of the parasite, which is assessed by immunofluorescence on the sporozoites and the liver stages of  Plasmodium falciparum  (but negative with respect to the erythrocytic stages); 
     high and specific lymphoproliferative responses against the 4 LSA-3 peptides, as well as the native T epitopes present at the surface of the sporozoites of  Plasmodium falciparum  and of  Plasmodium yoelii,  in which LSA-3 possesses a homologue (not yet characterized). 
     The B and T responses with respect to the native antigens are an important point since: 
     a) they prove that the synthetic molecules are indeed representative; 
     b) they signify that, at the time of infection, there are good prospects for obtaining an anamnestic secondary response; this is, in fact, what was observed in the chimpanzee Nuria at the time of the challenge. The importance of this observation is enhanced by the fact that the same secondary response was not obtained in respect of the other antigens such as LSA-1 and STARP. 
     2.5. Immunogenicity in Aotus: an owl monkey ( Aotus trivirgatus ) immunized with the 2 peptides LSA-3-NR1 and -RE and the 2 lipopeptides -NR2 and -CT1, and then restimulated with the recombinant proteins corresponding to 95% of the LSA-3 molecule and adsorbed on microspheres as described above, displays high and specific lymphoproliferative responses against the T epitopes present on these same peptides. 
     As regards the in vivo response of the different chimpanzees preimmunized in this way, the results underline the excellent immunogenicity (B and T) of LSA-3 in peptide, lipopeptide and recombinant form, and in all the animal models tested to date, namely 6/6 (outbred) chimpanzees and 1/1 Aotus, and in all the immunized mice (&gt;20). It may be noted that the results of the lipopeptide formulations (which can be used in man) were obtained by subcutaneous injection in the absence of any adjuvant. 
     EXAMPLE 3 
     Identification of CTL epitope 
     The method used to identify CTLs is the one described by Fidock et al., (1994), J. Immunol. 153: 190, or by Bottius et al., (1996), J. Immunol. 156: 2874-2884. 
     CTL (cytotoxic T lymphocyte) epitopes were identified in the peptides NR2, RE and CT1 by means of cytotoxicity tests performed on the PBMCs of the chimpanzees Dirk, Gerda, Nuria, Demi, Karlien and Iris described above. 
     In man, 8 additional CTL epitopes, 7 of them located in the 3′ non-repeat region, could be demonstrated on the PBMCs of individuals belonging to 3 different haplotypes (MHC class I-A2, -B8 and -B53) and living in a region where the disease is endemic (Gambia) (unpublished results). Furthermore, sequencing of the 2 B53-restricted CTL epitopes demonstrated a complete conservation of their nucleotide and peptide sequences in several strains from Kenya and from Gambia. 
     In total, we identified 11 CTL epitopes in the LSA-3 molecule, which is considerable. Moreover, 5/6 chimpanzees developed CTL responses against the peptide NR2 after immunization with the lipopeptide NR2 without adjuvant, which is a remarkable result for non-consanguineous animals. In addition, since the antibodies developed by Nuria did not display any inhibitory activity with respect to the invasion of  Plasmodium falciparum  sporozoites, it may be surmised that the observed protection depended on cellular responses, especially on the CTLs. 
     EXAMPLE 4 
     Comparison of the antibody titres before and after immunization with different peptides 4.1. Comparison of the antibody responses induced by different peptides in different immunized animals. 
     The method used is the one described in Behr et al., (1992), J. Immunol. 149: 3321. 
     The reactivity is expressed as an ELISA ratio, that is to say the optical density measured at 496 nanometers of the serum after immunization referred to the optical density of the same serum before immunization. The first column shows the animal immunized, the second column the immunogen received by the animal, the 3rd column shows the number of injections carried out as well as the support accompanying the peptide injected: RP denotes recombinant protein, RP/B denotes recombinant protein adsorbed on latex beads, P denotes peptide and LP lipopeptide. It should be pointed out, in addition, that the lipopeptides are injected in physiological saline, the peptides and the recombinant proteins are adsorbed on latex beads or in an emulsion with an adjuvant Montanide ISA-51. 
     
       
         
               
             
               
               
             
               
               
               
               
               
               
               
               
               
               
             
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
             
               
             
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
             
               
             
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
             
           
               
                 TABLE I 
               
             
             
               
                   
               
               
                 ANTIBODY REACTIVITY OF THE DIFFERENT PEPTIDES EXPRESSED AS AN ELISA RATIO 
               
             
          
           
               
                   
                 LSA-3 
               
             
          
           
               
                   
                 Injection 
                 LSA-1 
                 SALSA 
                 STARP 
                 LSA 
                 LSA 
                 LSA 
                 LSA 
                 R32T 
               
             
          
           
               
                   
                   
                 No. and 
                 LSA- 
                 LSA- 
                 LSA- 
                 LSA- 
                 SALSA 
                 SALSA 
                 STARP 
                 STARP 
                 -3- 
                 -3- 
                 -3- 
                 -3- 
                 and 
               
               
                 Chimpanzee 
                 Immunogen 
                 type 
                 REP 
                 J 
                 NR 
                 TER 
                 -1 
                 -2 
                 -M 
                 -R 
                 CT1 
                 NR1 
                 NRII 
                 REP 
                 32 
               
               
                   
               
             
          
           
               
                 Immunized animals 
               
             
          
           
               
                 DIRK 
                 LSA-3 and 
                 3RP(d) 
                 7.4 
                 9.0 
                 0.9 
                 0.8 
                 nd 
                 nd 
                 0.5 
                 0.7 
                 1.7 
                 1.0 
                 1.1 
                  8.8 
                 0.7 
               
               
                   
                 LSA-1 
                 3RP + 
                 20.0  
                 10.0  
                 0.1 
                 0.4 
                 0.2 
                 1.1 
                 1.0 
                 0.6 
                 1.0 
                 1.1 
                 3.1 
                 17.0 
                 0.8 
               
               
                   
                   
                 3 (P + LP) 
               
               
                 GERDA 
                 LSA-3 
                 3LP 
                 nd 
                 nd 
                 nd 
                 nd 
                 nd 
                 nd 
                 nd 
                 nd 
                 nd 
                 nd 
                 3.9 
                 nd 
                 0.6 
               
               
                   
                   
                 3LP + 
                 nd 
                 nd 
                 nd 
                 nd 
                 nd 
                 nd 
                 nd 
                 nd 
                 0.7 
                 1.1 
                 3.0 
                 12.3 
                 0.9 
               
               
                   
                   
                 3RP/B 
               
               
                 DEMI 
                 LSA-3 and 
                 2 (P + LP) 
                 8.0 
                 14.1  
                 0.7 
                 16.4  
                 0.6 
                 1.1 
                 nd 
                 nd 
                 0.7 
                 1.5 
                 11.7  
                 19.1 
                 0.7 
               
               
                   
                 LSA-1 
                 3 (P + LP) 
                 8.4 
                 14.5  
                 1.6 
                 21.5  
                 0.8 
                 0.2 
                 nd 
                 nd 
                 0.8 
                 5.1 
                 14.2  
                 20.7 
                 1.2 
               
               
                   
                   
                 3 (P + LP) + 
               
               
                   
                   
                 3RP/B 
               
               
                 KARLIEN 
                 LSA-3 and 
                 2 (P + LP) 
                 0.5 
                 1.2 
                 1.0 
                 1.0 
                 1.1 
                 2.1 
                 nd 
                 nd 
                 1.0 
                 3.6 
                 3.1 
                 10.3 
                 0.9 
               
               
                   
                 SALSA 
                 3 (P + LP) 
                 1.1 
                 0.2 
                 0.5 
                 0.2 
                 1.8 
                 2.5 
                 nd 
                 nd 
                 1.4 
                 4.7 
                 6.8 
                 14.1 
                 0.6 
               
               
                   
                   
                 3 (P + LP) + 
               
               
                   
                   
                 3RP/B 
               
               
                 IRIS 
                 LSA-3 and 
                 2 (P + LP) 
                 nd 
                 nd 
                 nd 
                 nd 
                 nd 
                 nd 
                 10.1  
                 15.9  
                 0.7 
                 2.4 
                 6.7 
                 12.5 
                 0.6 
               
               
                   
                 STARP 
                 3 (P + LP) 
                 nd 
                 nd 
                 nd 
                 nd 
                 nd 
                 nd 
                 10.5  
                 16.4  
                 1.3 
                 3.1 
                 6.8 
                 15.3 
                 0.5 
               
               
                   
                   
                 3 (P + LP) + 
               
               
                   
                   
                 3RP/B 
               
             
          
           
               
                 Unimmunized controls 
               
             
          
           
               
                 COR 
                 β-GAL 
                 3RP 
                 0.6 
                 0.7 
                 0.8 
                 0.9 
                 0.5 
                 1.0 
                 1.2 
                 0.8 
                 1.1 
                 1.0 
                 0.6 
                  1.1 
                 1.2 
               
               
                 PEER 
                 β-GAL 
                 6RP 
                 1.1 
                 0.8 
                 0.7 
                 0.9 
                 0.8 
                 1.2 
                 1.0 
                 0.9 
                 1.1 
                 0.6 
                 0.9 
                  0.9 
                 0.3 
               
               
                 BRAM 
                 GST 
                 2RP 
                 1.1 
                 0.6 
                 0.5 
                 1.1 
                 0.3 
                 0.8 
                 0.9 
                 1.2 
                 1.1 
                 0.3 
                 0.4 
                  0.7 
                 1.0 
               
               
                   
                   
                 3RP 
                 0.8 
                 0.3 
                 0.8 
                 1.3 
                 0.7 
                 1.2 
                 1.1 
                 1.2 
                 1.6 
                 0.2 
                 1.3 
                  0.6 
                 0.4 
               
               
                 FOUAD 
                 PBS 
                   
                 0.9 
                 0.5 
                 1.0 
                 0.6 
                 0.8 
                 1.3 
                 1.0 
                 0.3 
                 1.9 
                 1.3 
                 0.3 
                  0.2 
                 0.9 
               
               
                   
               
             
          
         
       
     
     4.2. Titre of the antibodies obtained: 
     Table II shows the antibody titres of the sera obtained in the chimpanzees by immunofluorescence on the native antigens present at the surface of the different stages (sporozoite, liver and blood) of  P.falciparum, P.yoelii  and  P.berghei.    
     
       
         
               
             
               
               
               
               
             
               
               
               
               
               
               
             
               
               
               
               
               
               
               
               
             
               
             
               
               
               
               
               
               
               
               
             
               
             
               
               
               
               
               
               
               
               
             
           
               
                 TABLE II 
               
             
             
               
                   
               
               
                 TITRE OF IMMUNOFLUORESCENT ANTIBODIES 
               
             
          
           
               
                   
                 
                   P. falciparum 
                 
                   
                 (17XL 
               
             
          
           
               
                   
                 SS 
                 LS 
                 BS 
                 
                   P. yoelii 
                 
                 and 17XNL) 
               
             
          
           
               
                 CHIMPANZEE 
                 Antigen 
                 (NF54) 
                 (NF54 and 73OX1) 
                 (150) 
                 SS 
                 LS 
                 BS 
               
               
                   
               
             
          
           
               
                 Immunized animals 
               
             
          
           
               
                 DIRK 
                 LSA-3 and 
                     800 
                     200 
                 −(&lt;100) 
                     200 
                     200 
                 −(&lt;100) 
               
               
                   
                 LSA-1 
               
               
                 GERDA 
                 LSA-3 
                     400 
                     200 
                 −(&lt;100) 
                     400 
                     200 
                 −(&lt;100) 
               
               
                 DEMI 
                 LSA-3 and 
                     100 
                     400 
                 −(&lt;100) 
               
               
                   
                 LSA-1 
               
               
                 KARLIEN 
                 LSA-3 and 
                     100 
                     200 
                 −(&lt;100) 
               
               
                   
                 SALSA 
               
               
                 IRIS 
                 LSA-3 and 
                     400 
                     100 
                 −(&lt;100) 
               
               
                   
                 STARP 
               
             
          
           
               
                 Control animals 
               
             
          
           
               
                 COR 
                 β-GAL 
                 −(&lt;100) 
                 −(&lt;100) 
                 −(&lt;100) 
                 −(&lt;100) 
                 −(&lt;100) 
                 −(&lt;100) 
               
               
                 BRAM 
                 GST 
                 −(&lt;100) 
                 −(&lt;100) 
                 −(&lt;100) 
                 −(&lt;100) 
                 −(&lt;100) 
                 −(&lt;100) 
               
               
                 FOUAD 
                 PBS 
                 −(&lt;100) 
                 −(&lt;100) 
                 −(&lt;100) 
                 −(&lt;100) 
                 −(&lt;100) 
                 −(&lt;100) 
               
               
                   
               
             
          
         
       
     
     4.3. Lymphoproliferative response of the PBMCs of the different chimpanzees after stimulation in vitro either with the different peptides or with the native antigens present at the surface of the sporozoites. This response was measured by incorporation of tritiated thymidine into PBMCs (peripheral blood cells) either after stimulation with the LSA-3 peptides (Table III) or after stimulation in vitro with sporozoites (Table IV). 
     
       
         
               
             
               
               
               
               
               
               
               
               
             
               
             
               
               
               
               
               
               
               
               
             
               
             
               
               
               
               
               
               
               
               
             
           
               
                 TABLE III 
               
             
             
               
                   
               
               
                 INCORPORATION OF TRITIATED THYMIDINE INTO 
               
               
                 PBMCs AFTER STIMULATION WITH THE LSA-3 PEPTIDES 
               
             
          
           
               
                 Chimpanzee 
                 Immunogen 
                 LSA-3-CT1 
                 LSA-3-NRI 
                 LSA-3-NRII 
                 LSA-3-REP 
                 MSP3-C (a) 
                 PPD (b) 
               
               
                   
               
             
          
           
               
                 Immunized animals 
               
             
          
           
               
                 DIRK 
                 LSA-3 and 
                 94,256 
                 27,125 
                 32,455  
                 69,321 
                 796 
                 89,338 
               
               
                   
                 LSA-1 
                 (4.0) 
                 (8.5) 
                 (10.7)  
                 (32.3) 
                 (1.0) 
                 (50.3) 
               
               
                 GERDA 
                 LSA-3 
                 13,359 
                  1,429 
                 13,236  
                 14,883 
                 485 
                 29,355 
               
               
                   
                   
                 (25.1)  
                 (2.8) 
                 (25.6)  
                 (28.6) 
                 (0.9) 
                 (132.3)  
               
               
                 DEMI 
                 LSA-3 and 
                 30,036 
                 17,221 
                 4,178 
                 52,301 
                 689 
                 167,277  
               
               
                   
                 LSA-1 
                 (46.8)  
                 (27.4)  
                 (7.3) 
                 (81.2) 
                 (1.1) 
                 (113.3)  
               
               
                 KARLIEN 
                 LSA-3 and 
                 30,025 
                 10,039 
                 18,365  
                 31,312 
                 575 
                 96.212 
               
               
                   
                 SALSA 
                 (36.4)  
                 (12.8)  
                 (23.1)  
                 (38.0) 
                 (0.7) 
                 (82.3) 
               
               
                 IRIS 
                 LSA-3 and 
                 53,312 
                 25,223 
                 6,458 
                 35,078 
                 799 
                 196,223  
               
               
                   
                 STARP 
                 (62.6)  
                 (34.8)  
                 (9.7) 
                 (47.5) 
                 (0.9) 
                 (62.3) 
               
             
          
           
               
                 Unimmunized animals 
               
             
          
           
               
                 COR 
                 β-GAL 
                  1,399 
                  2,599 
                 3,625 
                   786 
                 2,600   
                 19,395 
               
               
                   
                   
                 (0.6) 
                 (1.0) 
                 (1.3) 
                  (0.3) 
                 (1.1) 
                 (22.3) 
               
               
                 PEER 
                 β-GAL 
                  1,225 
                  1,369 
                 3,251 
                 2,960 
                 3,962   
                 59,399 
               
               
                   
                   
                 (0.2) 
                 (0.3) 
                 (1.2) 
                  (0.9) 
                 (1.5) 
                 (22.3) 
               
               
                 BRAM 
                 GST 
                  1,201 
                   509 
                 2,501 
                 2,659 
                 2,745   
                 39,399 
               
               
                   
                   
                 (0.4) 
                 (0.2) 
                 (0.7) 
                  (0.6) 
                 (0.7) 
                 (22.3) 
               
               
                 FOUAD 
                 PBS 
                  1,211 
                  1,310 
                   956 
                   688 
                 655 
                 136,258  
               
               
                   
                   
                 (1.2) 
                 (1.3) 
                 (0.9) 
                  (0.6) 
                 (0.5) 
                 (82.3) 
               
               
                   
               
               
                 (a) Control peptide from the MSP3 antigen in the blood  
               
               
                 (b) PPD = Purified protein derivative from Mycobacterium tuberculosis  
               
             
          
         
       
     
     
       
         
               
             
               
               
               
               
               
             
               
             
               
               
               
               
               
             
               
             
               
               
               
               
               
             
           
               
                 TABLE IV 
               
             
             
               
                   
               
               
                 INCORPORATION OF TRITIATED THYMIDINE INTO 
               
               
                 PBMCs AFTER STIMULATION IN VITRO WITH SPOROZOITES 
               
             
          
           
               
                   
                   
                 
                   P. falciparum 
                 
                 
                   P. yoelii 
                 
                 
                   P. berghei 
                 
               
               
                 Chimpanzee 
                 Antigen 
                 sporozoites 
                 sporozoites 
                 sporozoites 
               
               
                   
               
             
          
           
               
                 Immunized animals 
               
             
          
           
               
                 DIRK 
                 LSA-3 and LSA-1 
                 10,402 (12.1) 
                 5,552 (5.6) 
                 2,110 (2.0)   
               
               
                 GERDA 
                 LSA-3 
                 24,021 (20.5) 
                 18,228 (18.6) 
                 2,430 (0.7)   
               
               
                 DEMI 
                 LSA-3 and LSA-1 
                 2,111 (3.2) 
                   935 (1.4) 
                 214 (0.1) 
               
               
                 KARLIEN 
                 LSA-3 and SALSA 
                 4,402 (6.5) 
                 2,228 (3.6) 
                 914 (2.1) 
               
               
                 IRIS 
                 LSA-3 and STARP 
                  9,816 (14.2) 
                 5,304 (8.1) 
                 614 (2.0) 
               
             
          
           
               
                 Control animals 
               
             
          
           
               
                 BRAM 
                 GST 
                   245 (0.4) 
                 1,295 (1.6) 
                 514 (1.2) 
               
               
                 FOUAD 
                 PBS 
                   997 (1.5) 
                   828 (1.6) 
                 714 (1.1) 
               
               
                   
               
             
          
         
       
     
     The lymphoproliferative responses are shown as a counting difference in counts per minute (Δ CPM) between the number of counts obtained in the presence of antigen minus the number of counts in the absence of antigen. The figures in brackets show the stimulation indices, that is to say the ratio of the number of counts obtained in the presence of antigens to the number of counts obtained in the absence of antigens. 
     The results are considered to be positive when Δ CPM is greater than 1000 and when the stimulation index is greater than 3. 
     4.4. Comparison of the antibody responses of chimpanzee Nuria before and after immunization with different peptides 
     FIG. 5 depicts the amounts of immunoglobulins present in the serum of chimpanzee Nuria before and after immunization with the peptides 729NR1 and 729RE, and the lipopeptides 729NR2 and CT1. 
     This experiment shows the superiority as regards B immunity of the R antigen, most particularly when it is conjugated to a lipid residue. 
     FIG. 6 shows that the level of specific antibodies measured by ELISA against the peptide 729NR2 in mice immunized with either the peptide 729NRII or the lipopeptide 729NRII is markedly higher when the lipopeptide is used, irrespective of the species of mouse. 
     EXAMPLE 5 
     Lymphoproliferation of the PBMCs of an individual protected by injection of irradiated sporozoites against peptides originating from the LSA-1 and LSA-3 antigens 
     In eight human volunteers immunized by injection of irradiated sporozoites, anti-LSA-3 antibodies are found in each of the four individuals resistant to an infection by sporozoites; and none in the other four volunteers who developed a blood infection. 
     Furthermore, for the only one of these four protected individuals whose cells were accessible, the PBMCs were removed six months after the challenge infection and incubated in the presence of the peptides originating from the LSA-1 and LSA-3 antigens. 
     FIG. 10 depicts the results of lymphoproliferation of the PBMCs of an individual protected by injection of irradiated sporozoites against peptides originating from the LSA-1 and LSA-3 antigens. 
     Considerable lymphoproliferation was observed with each of the three peptides LSA-3 (NR1, NR2 and RE) but with none of the LSA-1 peptides. There was an especially high level of secretion of IFN-γ (100 IU/ml) after stimulation with the peptide NR1 and, to a lesser extent, with the peptide NR2 (IFN-γ: the cytokine having the strongest blocking effect on liver schizogony). 
     EXAMPLE 6 
     Effects of the antibodies against the LSA-3 peptides on the inhibition of the entry of sporozoites in mice 
     The techniques used to prepare the primary hepatocyte cultures, the sporozoites, the antibodies and the indirect fluorescence test are described in detail by S. Mellouk et al., Bulletin of the World Health Organization, 68: 52-59, 1990. Table V below compares the results obtained in immunofluorescence, either with antibodies against the fragment 679 or with antibodies obtained against fragments originating from other peptides. The left-hand column shows the number of schizonts detected after 48 h of culture in hepatocytes of Balb/c mice infected by  P.yoelii  and the right-hand column the same parameters after infection by  P.berghei.    
     
       
         
               
               
               
               
             
               
               
               
               
               
             
               
               
               
               
               
               
             
               
               
               
               
               
               
               
             
           
               
                   
                 TABLE V 
               
             
             
               
                   
                   
               
               
                   
                 
                   P. yoelii 
                 
                   
                 
                   P. berghei 
                 
               
             
          
           
               
                 Antibody 
                   
                 No. of LS at 48 h 
                   
                 No. of LS 
               
             
          
           
               
                 clones 
                 IFA 
                 a) 
                 b) 
                 IFA 
                 at 48 h 
               
               
                   
               
             
          
           
               
                 Control 
                   
                 88  
                 110  
                   
                 119 
                 108  
               
               
                 679 
                 ++ 
                   
                 0 
                 − 
                   
                 47 
               
               
                   
                 ++ 
                   
                 0 
                 − 
                   
                 ND 
               
               
                 679 
                 ++ 
                 1 
                   
                 − 
                 105 
               
               
                  679b 
                 ++ 
                 1 
                   
                 − 
                 133 
               
               
                  679c 
                 ++ 
                 1 
                   
                 − 
                  30 
               
               
                  32 
                 ++ 
                 8 
                   
                 ± 
                 103 
               
               
                 222 
                 + 
                   
                 5 
                 ± 
                   
                 26 
               
               
                 667 
                 ++ 
                 276  
                 143  
                 ND 
                 502 
               
               
                 362 
                 + 
                 3 
               
               
                 493 
                 ++ 
                 55  
                   
                 ND 
                 508 
               
               
                 α P.b. CSP Mab 
                   
                   
                 82 
                 +++ 
                   
                 30  
               
               
                 α P.y. CSP Mab 
                 +++ 
                   
                 171  
                   
                 138 
               
               
                   
               
             
          
         
       
     
     It is clearly apparent that the antibody against the peptide 679 has an almost complete inhibitory effect on the number of what they [sic] observed at 48 h in the liver cells. Likewise, FIG. 7 shows the inhibition of the sporozoite invasion of liver cells by hyperhuman [sic] sera obtained after immunization with different peptides and immunopurified against whole LSA-3. 
     As regards the protection of mice, the best results were obtained by immunization with the recombinants, or antigens prepared according to the invention, adsorbed on latex or polystyrene microspheres 0.5 μm in diameter: 
     3/3 mice are protected against an administration with 10 times the minimum infectious dose 
     3/3 mice are protected against the second challenge 
     2/3 mice are protected against the third challenge. 
     The microspheres used are Polybead® polystyrene microspheres (Polysciences, Inc.) 0.50 μm in diameter (ref. 07307) on which the recombinants or the peptides are adsorbed passively. In practice, in mice, per injection, 50 μg of antigens are brought into contact with 50 μl of microbeads; the exact amount of antigens adsorbed is not determined. In chimpanzees, the same procedure is performed with 200 μg of antigens and 200 μl of beads. 
     Furthermore, recently, the immunization of mice with the recombinant GST-3PC (corresponding to the non-repeat 3′ region from amino acid No. 869 to the stop codon at the 3′ end) has enabled sera to be obtained which react very strongly in immunofluorescence with  Plasmodium falciparum  sporozoites. This result is the first demonstration of the presence of one or several B epitopes in this region of the molecule. 
     EXAMPLE 7 
     Cytotoxicity test against the peptide 729NRII in the chimpanzee Gerda 
     The chimpanzee Gerda was immunized via the i.v. route with the lipopeptide 729NRII originating from the LSA-3 antigen. Blood is drawn 9 days after the 4th injection. The PBMCs were incubated in vitro with 5 μg/ml of the peptide 729NRII (addition of recombinant IL-2, 10 U/ml, on day 3). On day 15, the cytotoxic activity was studied against autologous blasts generated with PHA at a concentration of 0.5 μg/ml. The blasts were preincubated overnight with 5 μg/ml of the peptide 729NRII, and with a control peptide, namely RESA, or without a peptide. The peptides are not added during the test (8 hours). The number of targets per well is 5000. 
     PBMCs from Gerda incubated for the same period with 5 μg/ml of a control peptide or the peptide 729NRI (originating from the same antigen) do not bring about the lysis of autologous blasts preincubated or otherwise with the above peptides. 
     FIG. 8 shows the results obtained for an E/T (effector to target) ratio varying from 12 to 0.03. It is seen that the target cells presensitized with the peptide 729NRII are lysed in the presence of effector cells, indicating a cytotoxic T type immune response specific to this antigen. 
     The lipopeptide NRII injected via the i.v. route is capable, without adjuvant, of inducing a specific cytotoxic response. 
     EXAMPLE 8 
     Effect of the peptide NRI on interferon-γ production 
     Interferons have been shown to have an inhibitory activity in the development of  P.falciparum  in human hepatocytes in culture (Sylvie Mellouk et al., The Journal of Immunology, vol. 139 No. 12: 41-92, 41-95, 1987). The results obtained with the peptides of the invention are as follows: 
     The chimpanzee Gerda, immunized with the polypeptide NR2 and boosted with the recombinant DG729, carries PBMCs capable of secreting high levels of IFN-γ in the presence of the LSA-3 peptides, especially the peptide 729NRI. The result was confirmed in the chimpanzee Dirk, immunized with the same protein. The chimpanzee BRAN, an unimmunized control, does not show any interferon in the blood against the LSA-3 peptides. 
     
       
         
           
             29 
           
           
             1 
             6152 
             DNA 
             P. falciparum 
           
            1
atttatttat ttttattgtt ttatttcttt tttttcttta aattgtatat ttataaatat     60
tttaaaaagt tagaaaatga caaatagtaa ttacaaatca aataataaaa catataatga    120
aaataataat gaacaaataa ctaccatatt taatagaaca aatatgaatc cgataaaaaa    180
atgtcatatg agagaaaaaa taaataagta cttttttttg atcaaaattt tgacatgcac    240
cattttaata tgggctgtac aatatgataa taacgtaaga taaaaaacta aataataaat    300
ataaataaaa aaaaaaaaaa aaaaaaaaaa atcaactata tagtatgtat aatatatata    360
tatatatata tatatatata tatatatata tatttatttt tatttattta ttaatttttt    420
tttttttata ttatcttttt agtctgatat aaacaagagt tggaaaaaaa atacgtatgt    480
agataagaaa ttgaataaac tatttaacag aagtttagga gaatctcaag taaatggtga    540
attagctagt gaagaagtaa aggaaaaaat tcttgactta ttagaagaag gaaatacatt    600
aactgaaagt gtagatgata ataaaaattt agaagaagcc gaagatataa aggaaaatat    660
cttattaagt aatatagaag aaccaaaaga aaatattatt gacaatttat taaataatat    720
tggacaaaat tcagaaaaac aagaaagtgt atcagaaaat gtacaagtca gtgatgaact    780
ttttaatgaa ttattaaata gtgtagatgt taatggagaa gtaaaagaaa atattttgga    840
ggaaagtcaa gttaatgacg atatttttaa tagtttagta aaaagtgttc aacaagaaca    900
acaacacaat gttgaagaaa aagttgaaga aagtgtagaa gaaaatgacg aagaaagtgt    960
agaagaaaat gtagaagaaa atgtagaaga aaatgacgac ggaagtgtag cctcaagtgt   1020
tgaagaaagt atagcttcaa gtgttgatga aagtatagat tcaagtattg aagaaaatgt   1080
agctccaact gttgaagaaa tcgtagctcc aagtgttgta gaaagtgtgg ctccaagtgt   1140
tgaagaaagt gtagaagaaa atgttgaaga aagtgtagct gaaaatgttg aagaaagtgt   1200
agctgaaaat gttgaagaaa gtgtagctga aaatgttgaa gaaagtgtag ctgaaaatgt   1260
tgaagaaatc gtagctccaa ctgttgaaga aatcgtagct ccaactgttg aagaaattgt   1320
agctccaagt gttgtagaaa gtgtggctcc aagtgttgaa gaaagtgtag aagaaaatgt   1380
tgaagaaagt gtagctgaaa atgttgaaga aagtgtagct gaaaatgttg aagaaagtgt   1440
agctgaaaat gttgaagaaa gtgtagctga aaatgttgaa gaaagtgtag ctgaaaatgt   1500
tgaagaaatc gtagctccaa ctgttgaaga aatcgtagct ccaactgttg aagaaattgt   1560
agctccaagt gttgtagaaa gtgtggctcc aagtgttgaa gaaagtgtag aagaaaatgt   1620
tgaagaaagt gtagctgaaa atgttgaaga aagtgtagct gaaaatgttg aagaaagtgt   1680
agctgaaaat gttgaagaaa gtgtagctga aaatgttgaa gaaagtgtag ctgaaaatgt   1740
tgaagaaagt gtagctgaaa atgttgaaga aagtgtagct gaaaatgttg aagaaatcgt   1800
agctccaact gttgaagaaa tcgtagctcc aactgttgaa gaaattgtag ctccaagtgt   1860
tgtagaaagt gtggctccaa gtgttgaaga aagtgtagaa gaaaatgttg aagaaagtgt   1920
agctgaaaat gttgaagaaa gtgtagctga aaatgttgaa gaaagtgtag ctgaaaatgt   1980
tgaagaaagt gtagctgaaa atgttgaaga aatcgtagct ccaactgttg aagaaatcgt   2040
agctccaact gttgaagaaa ttgtagctcc aagtgttgta gaaagtgtgg ctccaagtgt   2100
tgaagaaagt gtagaagaaa atgttgaaga aagtgtagct gaaaatgttg aagaaagtgt   2160
agctgaaaat gttgaagaaa gtgtagctga aaatgttgaa gaaatcgtag ctccaactgt   2220
tgaagaaatc gtagctccaa ctgttgaaga aattgtagct ccaagtgttg tagaaagtgt   2280
ggctccaagt gttgaagaaa gtgtagaaga aaatgttgaa gaaagtgtag ctgaaaatgt   2340
tgaagaaagt gtagctgaaa atgttgaaga aagtgtagct gaaaatgttg aagaaagtgt   2400
agctgaaaat gttgaagaaa tcgtagctcc aactgttgaa gaaatcgtag ctccaactgt   2460
tgaagaaatt gtagctccaa gtgttgtaga aagtgtggct ccaagtgttg aagaaagtgt   2520
agaagaaaat gttgaagaaa gtgtagctga aaatgttgaa gaaagtgtag ctgaaaatgt   2580
tgaagaaagt gtagctgaaa atgttgaaga aagtgtagct ccaactgttg aagaaattgt   2640
agctccaagt gttgaagaaa gtgtagctcc aagtgttgaa gaaagtgttg ctgaaaacgt   2700
tgcaacaaat ttatcagaca atcttttaag taatttatta ggtggtatcg aaactgagga   2760
aataaaggac agtatattaa atgagataga agaagtaaaa gaaaatgtag tcaccacaat   2820
actagaaaac gtagaagaaa ctacagctga aagtgtaact acttttagta acatattaga   2880
ggagatacaa gaaaatacta ttactaatga tactatagag gaaaaattag aagaactcca   2940
cgaaaatgta ttaagtgccg ctttagaaaa tacccaaagt gaagaggaaa agaaagaagt   3000
aatagatgta attgaagaag taaaagaaga ggtcgctacc actttaatag aaactgtgga   3060
acaggcagaa gaaaagagcg caaatacaat tacggaaata tttgaaaatt tagaagaaaa   3120
tgcagtagaa agtaatgaaa atgttgcaga gaatttagag aaattaaacg aaactgtatt   3180
taatactgta ttagataaag tagaggaaac agtagaaatt agcggagaaa gtttagaaaa   3240
caatgaaatg gataaagcat tttttagtga aatatttgat aatgtaaaag gaatacaaga   3300
aaatttatta acaggtatgt ttcgaagtat agaaaccagt atagtaatcc aatcagaaga   3360
aaaggttgat ttgaatgaaa atgtggttag ttcgatttta gataatatag aaaatatgaa   3420
agaaggttta ttaaataaat tagaaaatat ttcaagtact gaaggtgttc aagaaactgt   3480
aactgaacat gtagaacaaa atgtatatgt ggatgttgat gttcctgcta tgaaagatca   3540
atttttagga atattaaatg aggcaggagg gttgaaagaa atgtttttta atttggaaga   3600
tgtatttaaa agtgaaagtg atgtaattac tgtagaagaa attaaggatg aaccggttca   3660
aaaagaggta gaaaaagaaa ctgttagtat tattgaagaa atggaagaaa atattgtaga   3720
tgtattagag gaagaaaaag aagatttaac agacaagatg atagatgcag tagaagaatc   3780
catagaaata tcttcagatt ctaaagaaga aactgaatct attaaagata aagaaaaaga   3840
tgtttcacta gttgttgaag aagttcaaga caatgatatg gatgaaagtg ttgagaaagt   3900
tttagaattg aaaaatatgg aagaggagtt aatgaaggat gctgttgaaa taaatgacat   3960
tactagcaaa cttattgaag aaactcaaga gttaaatgaa gtagaagcag atttaataaa   4020
agatatggaa aaattaaaag aattagaaaa agcattatca gaagattcta aagaaataat   4080
agatgcaaaa gatgatacat tagaaaaagt tattgaagag gaacatgata taacgacgac   4140
gttggatgaa gttgtagaat taaaagatgt cgaagaagac aagatcgaaa aagtatctga   4200
tttaaaagat cttgaagaag atatattaaa agaagtaaaa gaaatcaaag aacttgaaag   4260
tgaaatttta gaagattata aagaattaaa aactattgaa acagatattt tagaagagaa   4320
aaaagaaata gaaaaagatc attttgaaaa attcgaagaa gaagctgaag aaataaaaga   4380
tcttgaagca gatatattaa aagaagtatc ttcattagaa gttgaagaag aaaaaaaatt   4440
agaagaagta cacgaattaa aagaagaggt agaacatata ataagtggtg atgcgcatat   4500
aaaaggtttg gaagaagatg atttagaaga agtagatgat ttaaaaggaa gtatattaga   4560
catgttaaag ggagatatgg aattagggga tatggataag gaaagtttag aagatgtaac   4620
aacaaaactt ggagaaagag ttgaatcctt aaaagatgtt ttatctagtg cattaggcat   4680
ggatgaagaa caaatgaaaa caagaaaaaa agctcaaaga cctaagttgg aagaagtatt   4740
attaaaagaa gaggttaaag aagaaccaaa gaaaaaaata acaaaaaaga aagtaaggtt   4800
tgatattaag gataaggaac caaaagatga aatagtagaa gttgaaatga aagatgaaga   4860
tatagaagaa gatgtagaag aagatataga agaagatata gaagaagata aagttgaaga   4920
tatagatgaa gatatagatg aagatatagg tgaagacaaa gatgaagtta tagatttaat   4980
agtccaaaaa gagaaacgca ttgaaaaggt taaagcgaaa aagaaaaaat tagaaaaaaa   5040
agttgaagaa ggtgttagtg gtcttaaaaa acacgtagac gaagtaatga aatatgttca   5100
aaaaattgat aaagaagttg ataaagaagt atctaaagct ttagaatcaa aaaatgatgt   5160
tactaatgtt ttaaaacaaa atcaagattt ttttagtaaa gttaaaaact tcgtaaaaaa   5220
atataaagta tttgctgcac cattcatatc tgccgttgca gcatttgcat catatgtagt   5280
tgggttcttt acattttctt tattttcatc atgtgtaaca atagcttctt caacttactt   5340
attatcaaaa gttgacaaaa ctataaataa aaataaggag agaccgtttt attcatttgt   5400
atttgatatc tttaagaatt taaaacatta tttacaacaa atgaaagaaa aatttagtaa   5460
agaaaaaaat aataatgtaa tagaagtaac aaacaaagct gagaaaaaag gtaatgtaca   5520
ggtaacaaat aaaaccgaga aaacaactaa agttgataaa aataataaag taccgaaaaa   5580
aagaagaacg caaaaatcaa aataaaaaat tgcagaagag tgaaatgatt ggagcgaaca   5640
ataaaattaa tcgataaaaa atataaaaat gtatatatta tgtaaatata tataaataaa   5700
taaataaata catacatata tatatatata tatatgtatc tttttacaaa attttaaaat   5760
tttaaaattt atatatatta atatttatat ttttccatat ataattttat tttcaatatt   5820
ttatttttaa ttataaatgt tttttacaga gtttatgttt tttaattaat atatagattt   5880
ctgtaagaaa ctgtatatta ttcatacgat atatgtaata ttaattattt gtgttttatt   5940
aaaatttata ttatataata tatatatata tatatatgta tatatattag aagataaaaa   6000
tttagcttat tttgcttgtt atgcaaataa gctttttttt tttttttttt tttttttttc   6060
atataaacga tgtttaattt ttaattttta atattttata taaaatattt ttcctaaaaa   6120
aaaaaaaaat taaaaaaaac ttatatttcg aa                                 6152 
           
             2 
             5361 
             DNA 
             P. falciparum 
             
               CDS 
               (1)..(5361) 
             
           
            2
atg aca aat agt aat tac aaa tca aat aat aaa aca tat aat gaa aat       48
Met Thr Asn Ser Asn Tyr Lys Ser Asn Asn Lys Thr Tyr Asn Glu Asn
  1               5                  10                  15
aat aat gaa caa ata act acc ata ttt aat aga aca aat atg aat ccg       96
Asn Asn Glu Gln Ile Thr Thr Ile Phe Asn Arg Thr Asn Met Asn Pro
             20                  25                  30
ata aaa aaa tgt cat atg aga gaa aaa ata aat aag tac ttt ttt ttg      144
Ile Lys Lys Cys His Met Arg Glu Lys Ile Asn Lys Tyr Phe Phe Leu
         35                  40                  45
atc aaa att ttg aca tgc acc att tta ata tgg gct gta caa tat gat      192
Ile Lys Ile Leu Thr Cys Thr Ile Leu Ile Trp Ala Val Gln Tyr Asp
     50                  55                  60
aat aac tct gat ata aac aag agt tgg aaa aaa aat acg tat gta gat      240
Asn Asn Ser Asp Ile Asn Lys Ser Trp Lys Lys Asn Thr Tyr Val Asp
 65                  70                  75                  80
aag aaa ttg aat aaa cta ttt aac aga agt tta gga gaa tct caa gta      288
Lys Lys Leu Asn Lys Leu Phe Asn Arg Ser Leu Gly Glu Ser Gln Val
                 85                  90                  95
aat ggt gaa tta gct agt gaa gaa gta aag gaa aaa att ctt gac tta      336
Asn Gly Glu Leu Ala Ser Glu Glu Val Lys Glu Lys Ile Leu Asp Leu
            100                 105                 110
tta gaa gaa gga aat aca tta act gaa agt gta gat gat aat aaa aat      384
Leu Glu Glu Gly Asn Thr Leu Thr Glu Ser Val Asp Asp Asn Lys Asn
        115                 120                 125
tta gaa gaa gcc gaa gat ata aag gaa aat atc tta tta agt aat ata      432
Leu Glu Glu Ala Glu Asp Ile Lys Glu Asn Ile Leu Leu Ser Asn Ile
    130                 135                 140
gaa gaa cca aaa gaa aat att att gac aat tta tta aat aat att gga      480
Glu Glu Pro Lys Glu Asn Ile Ile Asp Asn Leu Leu Asn Asn Ile Gly
145                 150                 155                 160
caa aat tca gaa aaa caa gaa agt gta tca gaa aat gta caa gtc agt      528
Gln Asn Ser Glu Lys Gln Glu Ser Val Ser Glu Asn Val Gln Val Ser
                165                 170                 175
gat gaa ctt ttt aat gaa tta tta aat agt gta gat gtt aat gga gaa      576
Asp Glu Leu Phe Asn Glu Leu Leu Asn Ser Val Asp Val Asn Gly Glu
            180                 185                 190
gta aaa gaa aat att ttg gag gaa agt caa gtt aat gac gat att ttt      624
Val Lys Glu Asn Ile Leu Glu Glu Ser Gln Val Asn Asp Asp Ile Phe
        195                 200                 205
aat agt tta gta aaa agt gtt caa caa gaa caa caa cac aat gtt gaa      672
Asn Ser Leu Val Lys Ser Val Gln Gln Glu Gln Gln His Asn Val Glu
    210                 215                 220
gaa aaa gtt gaa gaa agt gta gaa gaa aat gac gaa gaa agt gta gaa      720
Glu Lys Val Glu Glu Ser Val Glu Glu Asn Asp Glu Glu Ser Val Glu
225                 230                 235                 240
gaa aat gta gaa gaa aat gta gaa gaa aat gac gac gga agt gta gcc      768
Glu Asn Val Glu Glu Asn Val Glu Glu Asn Asp Asp Gly Ser Val Ala
                245                 250                 255
tca agt gtt gaa gaa agt ata gct tca agt gtt gat gaa agt ata gat      816
Ser Ser Val Glu Glu Ser Ile Ala Ser Ser Val Asp Glu Ser Ile Asp
            260                 265                 270
tca agt att gaa gaa aat gta gct cca act gtt gaa gaa atc gta gct      864
Ser Ser Ile Glu Glu Asn Val Ala Pro Thr Val Glu Glu Ile Val Ala
        275                 280                 285
cca agt gtt gta gaa agt gtg gct cca agt gtt gaa gaa agt gta gaa      912
Pro Ser Val Val Glu Ser Val Ala Pro Ser Val Glu Glu Ser Val Glu
    290                 295                 300
gaa aat gtt gaa gaa agt gta gct gaa aat gtt gaa gaa agt gta gct      960
Glu Asn Val Glu Glu Ser Val Ala Glu Asn Val Glu Glu Ser Val Ala
305                 310                 315                 320
gaa aat gtt gaa gaa agt gta gct gaa aat gtt gaa gaa agt gta gct     1008
Glu Asn Val Glu Glu Ser Val Ala Glu Asn Val Glu Glu Ser Val Ala
                325                 330                 335
gaa aat gtt gaa gaa atc gta gct cca act gtt gaa gaa atc gta gct     1056
Glu Asn Val Glu Glu Ile Val Ala Pro Thr Val Glu Glu Ile Val Ala
            340                 345                 350
cca act gtt gaa gaa att gta gct cca agt gtt gta gaa agt gtg gct     1104
Pro Thr Val Glu Glu Ile Val Ala Pro Ser Val Val Glu Ser Val Ala
        355                 360                 365
cca agt gtt gaa gaa agt gta gaa gaa aat gtt gaa gaa agt gta gct     1152
Pro Ser Val Glu Glu Ser Val Glu Glu Asn Val Glu Glu Ser Val Ala
    370                 375                 380
gaa aat gtt gaa gaa agt gta gct gaa aat gtt gaa gaa agt gta gct     1200
Glu Asn Val Glu Glu Ser Val Ala Glu Asn Val Glu Glu Ser Val Ala
385                 390                 395                 400
gaa aat gtt gaa gaa agt gta gct gaa aat gtt gaa gaa agt gta gct     1248
Glu Asn Val Glu Glu Ser Val Ala Glu Asn Val Glu Glu Ser Val Ala
                405                 410                 415
gaa aat gtt gaa gaa atc gta gct cca act gtt gaa gaa atc gta gct     1296
Glu Asn Val Glu Glu Ile Val Ala Pro Thr Val Glu Glu Ile Val Ala
            420                 425                 430
cca act gtt gaa gaa att gta gct cca agt gtt gta gaa agt gtg gct     1344
Pro Thr Val Glu Glu Ile Val Ala Pro Ser Val Val Glu Ser Val Ala
        435                 440                 445
cca agt gtt gaa gaa agt gta gaa gaa aat gtt gaa gaa agt gta gct     1392
Pro Ser Val Glu Glu Ser Val Glu Glu Asn Val Glu Glu Ser Val Ala
    450                 455                 460
gaa aat gtt gaa gaa agt gta gct gaa aat gtt gaa gaa agt gta gct     1440
Glu Asn Val Glu Glu Ser Val Ala Glu Asn Val Glu Glu Ser Val Ala
465                 470                 475                 480
gaa aat gtt gaa gaa agt gta gct gaa aat gtt gaa gaa agt gta gct     1488
Glu Asn Val Glu Glu Ser Val Ala Glu Asn Val Glu Glu Ser Val Ala
                485                 490                 495
gaa aat gtt gaa gaa agt gta gct gaa aat gtt gaa gaa agt gta gct     1536
Glu Asn Val Glu Glu Ser Val Ala Glu Asn Val Glu Glu Ser Val Ala
            500                 505                 510
gaa aat gtt gaa gaa atc gta gct cca act gtt gaa gaa atc gta gct     1584
Glu Asn Val Glu Glu Ile Val Ala Pro Thr Val Glu Glu Ile Val Ala
        515                 520                 525
cca act gtt gaa gaa att gta gct cca agt gtt gta gaa agt gtg gct     1632
Pro Thr Val Glu Glu Ile Val Ala Pro Ser Val Val Glu Ser Val Ala
    530                 535                 540
cca agt gtt gaa gaa agt gta gaa gaa aat gtt gaa gaa agt gta gct     1680
Pro Ser Val Glu Glu Ser Val Glu Glu Asn Val Glu Glu Ser Val Ala
545                 550                 555                 560
gaa aat gtt gaa gaa agt gta gct gaa aat gtt gaa gaa agt gta gct     1728
Glu Asn Val Glu Glu Ser Val Ala Glu Asn Val Glu Glu Ser Val Ala
                565                 570                 575
gaa aat gtt gaa gaa agt gta gct gaa aat gtt gaa gaa atc gta gct     1776
Glu Asn Val Glu Glu Ser Val Ala Glu Asn Val Glu Glu Ile Val Ala
            580                 585                 590
cca act gtt gaa gaa atc gta gct cca act gtt gaa gaa att gta gct     1824
Pro Thr Val Glu Glu Ile Val Ala Pro Thr Val Glu Glu Ile Val Ala
        595                 600                 605
cca agt gtt gta gaa agt gtg gct cca agt gtt gaa gaa agt gta gaa     1872
Pro Ser Val Val Glu Ser Val Ala Pro Ser Val Glu Glu Ser Val Glu
    610                 615                 620
gaa aat gtt gaa gaa agt gta gct gaa aat gtt gaa gaa agt gta gct     1920
Glu Asn Val Glu Glu Ser Val Ala Glu Asn Val Glu Glu Ser Val Ala
625                 630                 635                 640
gaa aat gtt gaa gaa agt gta gct gaa aat gtt gaa gaa atc gta gct     1968
Glu Asn Val Glu Glu Ser Val Ala Glu Asn Val Glu Glu Ile Val Ala
                645                 650                 655
cca act gtt gaa gaa atc gta gct cca act gtt gaa gaa att gta gct     2016
Pro Thr Val Glu Glu Ile Val Ala Pro Thr Val Glu Glu Ile Val Ala
            660                 665                 670
cca agt gtt gta gaa agt gtg gct cca agt gtt gaa gaa agt gta gaa     2064
Pro Ser Val Val Glu Ser Val Ala Pro Ser Val Glu Glu Ser Val Glu
        675                 680                 685
gaa aat gtt gaa gaa agt gta gct gaa aat gtt gaa gaa agt gta gct     2112
Glu Asn Val Glu Glu Ser Val Ala Glu Asn Val Glu Glu Ser Val Ala
    690                 695                 700
gaa aat gtt gaa gaa agt gta gct gaa aat gtt gaa gaa agt gta gct     2160
Glu Asn Val Glu Glu Ser Val Ala Glu Asn Val Glu Glu Ser Val Ala
705                 710                 715                 720
gaa aat gtt gaa gaa atc gta gct cca act gtt gaa gaa atc gta gct     2208
Glu Asn Val Glu Glu Ile Val Ala Pro Thr Val Glu Glu Ile Val Ala
                725                 730                 735
cca act gtt gaa gaa att gta gct cca agt gtt gta gaa agt gtg gct     2256
Pro Thr Val Glu Glu Ile Val Ala Pro Ser Val Val Glu Ser Val Ala
            740                 745                 750
cca agt gtt gaa gaa agt gta gaa gaa aat gtt gaa gaa agt gta gct     2304
Pro Ser Val Glu Glu Ser Val Glu Glu Asn Val Glu Glu Ser Val Ala
        755                 760                 765
gaa aat gtt gaa gaa agt gta gct gaa aat gtt gaa gaa agt gta gct     2352
Glu Asn Val Glu Glu Ser Val Ala Glu Asn Val Glu Glu Ser Val Ala
    770                 775                 780
gaa aat gtt gaa gaa agt gta gct cca act gtt gaa gaa att gta gct     2400
Glu Asn Val Glu Glu Ser Val Ala Pro Thr Val Glu Glu Ile Val Ala
785                 790                 795                 800
cca agt gtt gaa gaa agt gta gct cca agt gtt gaa gaa agt gtt gct     2448
Pro Ser Val Glu Glu Ser Val Ala Pro Ser Val Glu Glu Ser Val Ala
                805                 810                 815
gaa aac gtt gca aca aat tta tca gac aat ctt tta agt aat tta tta     2496
Glu Asn Val Ala Thr Asn Leu Ser Asp Asn Leu Leu Ser Asn Leu Leu
            820                 825                 830
ggt ggt atc gaa act gag gaa ata aag gac agt ata tta aat gag ata     2544
Gly Gly Ile Glu Thr Glu Glu Ile Lys Asp Ser Ile Leu Asn Glu Ile
        835                 840                 845
gaa gaa gta aaa gaa aat gta gtc acc aca ata cta gaa aac gta gaa     2592
Glu Glu Val Lys Glu Asn Val Val Thr Thr Ile Leu Glu Asn Val Glu
    850                 855                 860
gaa act aca gct gaa agt gta act act ttt agt aac ata tta gag gag     2640
Glu Thr Thr Ala Glu Ser Val Thr Thr Phe Ser Asn Ile Leu Glu Glu
865                 870                 875                 880
ata caa gaa aat act att act aat gat act ata gag gaa aaa tta gaa     2688
Ile Gln Glu Asn Thr Ile Thr Asn Asp Thr Ile Glu Glu Lys Leu Glu
                885                 890                 895
gaa ctc cac gaa aat gta tta agt gcc gct tta gaa aat acc caa agt     2736
Glu Leu His Glu Asn Val Leu Ser Ala Ala Leu Glu Asn Thr Gln Ser
            900                 905                 910
gaa gag gaa aag aaa gaa gta ata gat gta att gaa gaa gta aaa gaa     2784
Glu Glu Glu Lys Lys Glu Val Ile Asp Val Ile Glu Glu Val Lys Glu
        915                 920                 925
gag gtc gct acc act tta ata gaa act gtg gaa cag gca gaa gaa aag     2832
Glu Val Ala Thr Thr Leu Ile Glu Thr Val Glu Gln Ala Glu Glu Lys
    930                 935                 940
agc gca aat aca att acg gaa ata ttt gaa aat tta gaa gaa aat gca     2880
Ser Ala Asn Thr Ile Thr Glu Ile Phe Glu Asn Leu Glu Glu Asn Ala
945                 950                 955                 960
gta gaa agt aat gaa aat gtt gca gag aat tta gag aaa tta aac gaa     2928
Val Glu Ser Asn Glu Asn Val Ala Glu Asn Leu Glu Lys Leu Asn Glu
                965                 970                 975
act gta ttt aat act gta tta gat aaa gta gag gaa aca gta gaa att     2976
Thr Val Phe Asn Thr Val Leu Asp Lys Val Glu Glu Thr Val Glu Ile
            980                 985                 990
agc gga gaa agt tta gaa aac aat gaa atg gat aaa gca ttt ttt agt     3024
Ser Gly Glu Ser Leu Glu Asn Asn Glu Met Asp Lys Ala Phe Phe Ser
        995                1000                1005
gaa ata ttt gat aat gta aaa gga ata caa gaa aat tta tta aca ggt     3072
Glu Ile Phe Asp Asn Val Lys Gly Ile Gln Glu Asn Leu Leu Thr Gly
   1010                1015                1020
atg ttt cga agt ata gaa acc agt ata gta atc caa tca gaa gaa aag     3120
Met Phe Arg Ser Ile Glu Thr Ser Ile Val Ile Gln Ser Glu Glu Lys
1025               1030                1035                1040
gtt gat ttg aat gaa aat gtg gtt agt tcg att tta gat aat ata gaa     3168
Val Asp Leu Asn Glu Asn Val Val Ser Ser Ile Leu Asp Asn Ile Glu
               1045                1050                1055
aat atg aaa gaa ggt tta tta aat aaa tta gaa aat att tca agt act     3216
Asn Met Lys Glu Gly Leu Leu Asn Lys Leu Glu Asn Ile Ser Ser Thr
           1060                1065                1070
gaa ggt gtt caa gaa act gta act gaa cat gta gaa caa aat gta tat     3264
Glu Gly Val Gln Glu Thr Val Thr Glu His Val Glu Gln Asn Val Tyr
       1075                1080                1085
gtg gat gtt gat gtt cct gct atg aaa gat caa ttt tta gga ata tta     3312
Val Asp Val Asp Val Pro Ala Met Lys Asp Gln Phe Leu Gly Ile Leu
   1090                1095                1100
aat gag gca gga ggg ttg aaa gaa atg ttt ttt aat ttg gaa gat gta     3360
Asn Glu Ala Gly Gly Leu Lys Glu Met Phe Phe Asn Leu Glu Asp Val
1105               1110                1115                1120
ttt aaa agt gaa agt gat gta att act gta gaa gaa att aag gat gaa     3408
Phe Lys Ser Glu Ser Asp Val Ile Thr Val Glu Glu Ile Lys Asp Glu
               1125                1130                1135
ccg gtt caa aaa gag gta gaa aaa gaa act gtt agt att att gaa gaa     3456
Pro Val Gln Lys Glu Val Glu Lys Glu Thr Val Ser Ile Ile Glu Glu
           1140                1145                1150
atg gaa gaa aat att gta gat gta tta gag gaa gaa aaa gaa gat tta     3504
Met Glu Glu Asn Ile Val Asp Val Leu Glu Glu Glu Lys Glu Asp Leu
       1155                1160                1165
aca gac aag atg ata gat gca gta gaa gaa tcc ata gaa ata tct tca     3552
Thr Asp Lys Met Ile Asp Ala Val Glu Glu Ser Ile Glu Ile Ser Ser
   1170                1175                1180
gat tct aaa gaa gaa act gaa tct att aaa gat aaa gaa aaa gat gtt     3600
Asp Ser Lys Glu Glu Thr Glu Ser Ile Lys Asp Lys Glu Lys Asp Val
1185               1190                1195                1200
tca cta gtt gtt gaa gaa gtt caa gac aat gat atg gat gaa agt gtt     3648
Ser Leu Val Val Glu Glu Val Gln Asp Asn Asp Met Asp Glu Ser Val
               1205                1210                1215
gag aaa gtt tta gaa ttg aaa aat atg gaa gag gag tta atg aag gat     3696
Glu Lys Val Leu Glu Leu Lys Asn Met Glu Glu Glu Leu Met Lys Asp
           1220                1225                1230
gct gtt gaa ata aat gac att act agc aaa ctt att gaa gaa act caa     3744
Ala Val Glu Ile Asn Asp Ile Thr Ser Lys Leu Ile Glu Glu Thr Gln
       1235                1240                1245
gag tta aat gaa gta gaa gca gat tta ata aaa gat atg gaa aaa tta     3792
Glu Leu Asn Glu Val Glu Ala Asp Leu Ile Lys Asp Met Glu Lys Leu
   1250                1255                1260
aaa gaa tta gaa aaa gca tta tca gaa gat tct aaa gaa ata ata gat     3840
Lys Glu Leu Glu Lys Ala Leu Ser Glu Asp Ser Lys Glu Ile Ile Asp
1265               1270                1275                1280
gca aaa gat gat aca tta gaa aaa gtt att gaa gag gaa cat gat ata     3888
Ala Lys Asp Asp Thr Leu Glu Lys Val Ile Glu Glu Glu His Asp Ile
               1285                1290                1295
acg acg acg ttg gat gaa gtt gta gaa tta aaa gat gtc gaa gaa gac     3936
Thr Thr Thr Leu Asp Glu Val Val Glu Leu Lys Asp Val Glu Glu Asp
           1300                1305                1310
aag atc gaa aaa gta tct gat tta aaa gat ctt gaa gaa gat ata tta     3984
Lys Ile Glu Lys Val Ser Asp Leu Lys Asp Leu Glu Glu Asp Ile Leu
       1315                1320                1325
aaa gaa gta aaa gaa atc aaa gaa ctt gaa agt gaa att tta gaa gat     4032
Lys Glu Val Lys Glu Ile Lys Glu Leu Glu Ser Glu Ile Leu Glu Asp
   1330                1335                1340
tat aaa gaa tta aaa act att gaa aca gat att tta gaa gag aaa aaa     4080
Tyr Lys Glu Leu Lys Thr Ile Glu Thr Asp Ile Leu Glu Glu Lys Lys
1345               1350                1355                1360
gaa ata gaa aaa gat cat ttt gaa aaa ttc gaa gaa gaa gct gaa gaa     4128
Glu Ile Glu Lys Asp His Phe Glu Lys Phe Glu Glu Glu Ala Glu Glu
               1365                1370                1375
ata aaa gat ctt gaa gca gat ata tta aaa gaa gta tct tca tta gaa     4176
Ile Lys Asp Leu Glu Ala Asp Ile Leu Lys Glu Val Ser Ser Leu Glu
           1380                1385                1390
gtt gaa gaa gaa aaa aaa tta gaa gaa gta cac gaa tta aaa gaa gag     4224
Val Glu Glu Glu Lys Lys Leu Glu Glu Val His Glu Leu Lys Glu Glu
       1395                1400                1405
gta gaa cat ata ata agt ggt gat gcg cat ata aaa ggt ttg gaa gaa     4272
Val Glu His Ile Ile Ser Gly Asp Ala His Ile Lys Gly Leu Glu Glu
   1410                1415                1420
gat gat tta gaa gaa gta gat gat tta aaa gga agt ata tta gac atg     4320
Asp Asp Leu Glu Glu Val Asp Asp Leu Lys Gly Ser Ile Leu Asp Met
1425               1430                1435                1440
tta aag gga gat atg gaa tta ggg gat atg gat aag gaa agt tta gaa     4368
Leu Lys Gly Asp Met Glu Leu Gly Asp Met Asp Lys Glu Ser Leu Glu
               1445                1450                1455
gat gta aca aca aaa ctt gga gaa aga gtt gaa tcc tta aaa gat gtt     4416
Asp Val Thr Thr Lys Leu Gly Glu Arg Val Glu Ser Leu Lys Asp Val
           1460                1465                1470
tta tct agt gca tta ggc atg gat gaa gaa caa atg aaa aca aga aaa     4464
Leu Ser Ser Ala Leu Gly Met Asp Glu Glu Gln Met Lys Thr Arg Lys
       1475                1480                1485
aaa gct caa aga cct aag ttg gaa gaa gta tta tta aaa gaa gag gtt     4512
Lys Ala Gln Arg Pro Lys Leu Glu Glu Val Leu Leu Lys Glu Glu Val
   1490                1495                1500
aaa gaa gaa cca aag aaa aaa ata aca aaa aag aaa gta agg ttt gat     4560
Lys Glu Glu Pro Lys Lys Lys Ile Thr Lys Lys Lys Val Arg Phe Asp
1505               1510                1515                1520
att aag gat aag gaa cca aaa gat gaa ata gta gaa gtt gaa atg aaa     4608
Ile Lys Asp Lys Glu Pro Lys Asp Glu Ile Val Glu Val Glu Met Lys
               1525                1530                1535
gat gaa gat ata gaa gaa gat gta gaa gaa gat ata gaa gaa gat ata     4656
Asp Glu Asp Ile Glu Glu Asp Val Glu Glu Asp Ile Glu Glu Asp Ile
           1540                1545                1550
gaa gaa gat aaa gtt gaa gat ata gat gaa gat ata gat gaa gat ata     4704
Glu Glu Asp Lys Val Glu Asp Ile Asp Glu Asp Ile Asp Glu Asp Ile
       1555                1560                1565
ggt gaa gac aaa gat gaa gtt ata gat tta ata gtc caa aaa gag aaa     4752
Gly Glu Asp Lys Asp Glu Val Ile Asp Leu Ile Val Gln Lys Glu Lys
   1570                1575                1580
cgc att gaa aag gtt aaa gcg aaa aag aaa aaa tta gaa aaa aaa gtt     4800
Arg Ile Glu Lys Val Lys Ala Lys Lys Lys Lys Leu Glu Lys Lys Val
1585               1590                1595                1600
gaa gaa ggt gtt agt ggt ctt aaa aaa cac gta gac gaa gta atg aaa     4848
Glu Glu Gly Val Ser Gly Leu Lys Lys His Val Asp Glu Val Met Lys
               1605                1610                1615
tat gtt caa aaa att gat aaa gaa gtt gat aaa gaa gta tct aaa gct     4896
Tyr Val Gln Lys Ile Asp Lys Glu Val Asp Lys Glu Val Ser Lys Ala
           1620                1625                1630
tta gaa tca aaa aat gat gtt act aat gtt tta aaa caa aat caa gat     4944
Leu Glu Ser Lys Asn Asp Val Thr Asn Val Leu Lys Gln Asn Gln Asp
       1635                1640                1645
ttt ttt agt aaa gtt aaa aac ttc gta aaa aaa tat aaa gta ttt gct     4992
Phe Phe Ser Lys Val Lys Asn Phe Val Lys Lys Tyr Lys Val Phe Ala
   1650                1655                1660
gca cca ttc ata tct gcc gtt gca gca ttt gca tca tat gta gtt ggg     5040
Ala Pro Phe Ile Ser Ala Val Ala Ala Phe Ala Ser Tyr Val Val Gly
1665               1670                1675                1680
ttc ttt aca ttt tct tta ttt tca tca tgt gta aca ata gct tct tca     5088
Phe Phe Thr Phe Ser Leu Phe Ser Ser Cys Val Thr Ile Ala Ser Ser
               1685                1690                1695
act tac tta tta tca aaa gtt gac aaa act ata aat aaa aat aag gag     5136
Thr Tyr Leu Leu Ser Lys Val Asp Lys Thr Ile Asn Lys Asn Lys Glu
           1700                1705                1710
aga ccg ttt tat tca ttt gta ttt gat atc ttt aag aat tta aaa cat     5184
Arg Pro Phe Tyr Ser Phe Val Phe Asp Ile Phe Lys Asn Leu Lys His
       1715                1720                1725
tat tta caa caa atg aaa gaa aaa ttt agt aaa gaa aaa aat aat aat     5232
Tyr Leu Gln Gln Met Lys Glu Lys Phe Ser Lys Glu Lys Asn Asn Asn
   1730                1735                1740
gta ata gaa gta aca aac aaa gct gag aaa aaa ggt aat gta cag gta     5280
Val Ile Glu Val Thr Asn Lys Ala Glu Lys Lys Gly Asn Val Gln Val
1745               1750                1755                1760
aca aat aaa acc gag aaa aca act aaa gtt gat aaa aat aat aaa gta     5328
Thr Asn Lys Thr Glu Lys Thr Thr Lys Val Asp Lys Asn Asn Lys Val
               1765                1770                1775
ccg aaa aaa aga aga acg caa aaa tca aaa taa                         5361
Pro Lys Lys Arg Arg Thr Gln Lys Ser Lys
           1780                1785 
           
             3 
             1891 
             DNA 
             P. falciparum 
             
               CDS 
               (2)..(1891) 
             
           
            3
t aca tta act gaa agt gta gat gat aat aaa aat tta gaa gaa gcc gaa     49
  Thr Leu Thr Glu Ser Val Asp Asp Asn Lys Asn Leu Glu Glu Ala Glu
    1               5                  10                  15
gat ata aag gaa aat atc tta tta agt aat ata gaa gaa cca aaa gaa       97
Asp Ile Lys Glu Asn Ile Leu Leu Ser Asn Ile Glu Glu Pro Lys Glu
             20                  25                  30
aat att att gac aat tta tta aat aat att gga caa aat tca gaa aaa      145
Asn Ile Ile Asp Asn Leu Leu Asn Asn Ile Gly Gln Asn Ser Glu Lys
         35                  40                  45
caa gaa agt gta tca gaa aat gta caa gtc agt gat gaa ctt ttt aat      193
Gln Glu Ser Val Ser Glu Asn Val Gln Val Ser Asp Glu Leu Phe Asn
     50                  55                  60
gaa tta tta aat agt gta gat gtt aat gga gaa gta aaa gaa aat att      241
Glu Leu Leu Asn Ser Val Asp Val Asn Gly Glu Val Lys Glu Asn Ile
 65                  70                  75                  80
ttg gag gaa agt caa gtt aat gac gat att ttt aat agt tta gta aaa      289
Leu Glu Glu Ser Gln Val Asn Asp Asp Ile Phe Asn Ser Leu Val Lys
                 85                  90                  95
agt gtt caa caa gaa caa caa cac aat gtt gaa gaa aaa gtt gaa gaa      337
Ser Val Gln Gln Glu Gln Gln His Asn Val Glu Glu Lys Val Glu Glu
            100                 105                 110
agt gta gaa gaa aat gac gaa gaa agt gta gaa gaa aat gta gaa gaa      385
Ser Val Glu Glu Asn Asp Glu Glu Ser Val Glu Glu Asn Val Glu Glu
        115                 120                 125
aat gta gaa gaa aat gac gac gga agt gta gcc tca agt gtt gaa gaa      433
Asn Val Glu Glu Asn Asp Asp Gly Ser Val Ala Ser Ser Val Glu Glu
    130                 135                 140
agt ata gct tca agt gtt gat gaa agt ata gat tca agt att gaa gaa      481
Ser Ile Ala Ser Ser Val Asp Glu Ser Ile Asp Ser Ser Ile Glu Glu
145                 150                 155                 160
aat gta gct cca act gtt gaa gaa atc gta gct cca act gtt gaa gaa      529
Asn Val Ala Pro Thr Val Glu Glu Ile Val Ala Pro Thr Val Glu Glu
                165                 170                 175
att gta gct cca agt gtt gta gaa agt gtg gct cca agt gtt gaa gaa      577
Ile Val Ala Pro Ser Val Val Glu Ser Val Ala Pro Ser Val Glu Glu
            180                 185                 190
agt gta gct cca agt gtt gaa gaa agt gta gct gaa aat gtt gaa gaa      625
Ser Val Ala Pro Ser Val Glu Glu Ser Val Ala Glu Asn Val Glu Glu
        195                 200                 205
agt gta gct gaa aat gtt gaa gaa atc gta gct cca agt gtt gaa gaa      673
Ser Val Ala Glu Asn Val Glu Glu Ile Val Ala Pro Ser Val Glu Glu
    210                 215                 220
agt gta gct gaa aat gtt gaa gaa agt gta gct gaa aat gtt gaa gaa      721
Ser Val Ala Glu Asn Val Glu Glu Ser Val Ala Glu Asn Val Glu Glu
225                 230                 235                 240
agt gta gct gaa aat gtt gaa gaa agt gta gct gaa aat gtt gaa gaa      769
Ser Val Ala Glu Asn Val Glu Glu Ser Val Ala Glu Asn Val Glu Glu
                245                 250                 255
agt gta gct gaa aat gtt gaa gaa atc gta gct cca act gtt gaa gaa      817
Ser Val Ala Glu Asn Val Glu Glu Ile Val Ala Pro Thr Val Glu Glu
            260                 265                 270
agt gta gct cca act gtt gaa gaa att gta gct cca act gtt gaa gaa      865
Ser Val Ala Pro Thr Val Glu Glu Ile Val Ala Pro Thr Val Glu Glu
        275                 280                 285
agt gta gct cca act gtt gaa gaa att gta gtt cca agt gtt gaa gaa      913
Ser Val Ala Pro Thr Val Glu Glu Ile Val Val Pro Ser Val Glu Glu
    290                 295                 300
agt gta gct cca agt gtt gaa gaa agt gta gct gaa aat gtt gaa gaa      961
Ser Val Ala Pro Ser Val Glu Glu Ser Val Ala Glu Asn Val Glu Glu
305                 310                 315                 320
agt gta gct gaa aat gtt gaa gaa agt gta gct gaa aat gtt gaa gaa     1009
Ser Val Ala Glu Asn Val Glu Glu Ser Val Ala Glu Asn Val Glu Glu
                325                 330                 335
agt gta gct gaa aat gtt gaa gaa agt gta gct gaa aat gtt gaa gaa     1057
Ser Val Ala Glu Asn Val Glu Glu Ser Val Ala Glu Asn Val Glu Glu
            340                 345                 350
atc gta gct cca agt gtt gaa gaa atc gta gct cca act gtt gaa gaa     1105
Ile Val Ala Pro Ser Val Glu Glu Ile Val Ala Pro Thr Val Glu Glu
        355                 360                 365
agt gtt gct gaa aac gtt gca aca aat tta tca gac aat ctt tta agt     1153
Ser Val Ala Glu Asn Val Ala Thr Asn Leu Ser Asp Asn Leu Leu Ser
    370                 375                 380
aat tta tta ggt ggt atc gaa act gag gaa ata aag gac agt ata tta     1201
Asn Leu Leu Gly Gly Ile Glu Thr Glu Glu Ile Lys Asp Ser Ile Leu
385                 390                 395                 400
aat gag ata gaa gaa gta aaa gaa aat gta gtc acc aca ata cta gaa     1249
Asn Glu Ile Glu Glu Val Lys Glu Asn Val Val Thr Thr Ile Leu Glu
                405                 410                 415
aaa gta gaa gaa act aca gct gaa agt gta act act ttt agt aat ata     1297
Lys Val Glu Glu Thr Thr Ala Glu Ser Val Thr Thr Phe Ser Asn Ile
            420                 425                 430
tta gag gag ata caa gaa aat act att act aat gat act ata gag gaa     1345
Leu Glu Glu Ile Gln Glu Asn Thr Ile Thr Asn Asp Thr Ile Glu Glu
        435                 440                 445
aaa tta gaa gaa ctc cac gaa aat gta tta agt gcc gct tta gaa aat     1393
Lys Leu Glu Glu Leu His Glu Asn Val Leu Ser Ala Ala Leu Glu Asn
    450                 455                 460
acc caa agt gaa gag gaa aag aaa gaa gta ata gat gta att gaa gaa     1441
Thr Gln Ser Glu Glu Glu Lys Lys Glu Val Ile Asp Val Ile Glu Glu
465                 470                 475                 480
gta aaa gaa gag gtc gct acc act tta ata gaa act gtg gaa cag gca     1489
Val Lys Glu Glu Val Ala Thr Thr Leu Ile Glu Thr Val Glu Gln Ala
                485                 490                 495
gaa gaa gag agc gaa agt aca att acg gaa ata ttt gaa aat tta gaa     1537
Glu Glu Glu Ser Glu Ser Thr Ile Thr Glu Ile Phe Glu Asn Leu Glu
            500                 505                 510
gaa aat gca gta gaa agt aat gaa aaa gtt gca gag aat tta gag aaa     1585
Glu Asn Ala Val Glu Ser Asn Glu Lys Val Ala Glu Asn Leu Glu Lys
        515                 520                 525
tta aac gaa act gta ttt aat act gta tta gat aaa gta gag gaa aca     1633
Leu Asn Glu Thr Val Phe Asn Thr Val Leu Asp Lys Val Glu Glu Thr
    530                 535                 540
gta gaa att agc gga gaa agt tta gaa aac aat gaa atg gat aaa gca     1681
Val Glu Ile Ser Gly Glu Ser Leu Glu Asn Asn Glu Met Asp Lys Ala
545                 550                 555                 560
ttt ttt agt gaa ata ttt gat aat gta aaa gga ata caa gaa aat tta     1729
Phe Phe Ser Glu Ile Phe Asp Asn Val Lys Gly Ile Gln Glu Asn Leu
                565                 570                 575
tta aca ggt atg ttt cga agt ata gaa acc agt ata gta atc caa tca     1777
Leu Thr Gly Met Phe Arg Ser Ile Glu Thr Ser Ile Val Ile Gln Ser
            580                 585                 590
gaa gaa aag gtt gat ttg aat gaa aat gtg gtt agt tcg att tta gat     1825
Glu Glu Lys Val Asp Leu Asn Glu Asn Val Val Ser Ser Ile Leu Asp
        595                 600                 605
aat ata gaa aat atg aaa gaa ggt tta tta aat aaa tta gaa aat att     1873
Asn Ile Glu Asn Met Lys Glu Gly Leu Leu Asn Lys Leu Glu Asn Ile
    610                 615                 620
tca agt act gaa ggc gaa                                             1891
Ser Ser Thr Glu Gly Glu
625                 630 
           
             4 
             28 
             DNA 
             Artificial Sequence 
             
               Description of Artificial SequencePrimer 
             
           
            4
gtgatgaact ttttaatgaa ttattaaa                                        28 
           
             5 
             29 
             DNA 
             Artificial Sequence 
             
               Description of Artificial SequencePrimer 
             
           
            5
tgttgttctt gttgaacact ttttactaa                                       29 
           
             6 
             25 
             DNA 
             Artificial Sequence 
             
               Description of Artificial SequencePrimer 
             
           
            6
ggtatcgaaa ctgaggaaat aaagg                                           25 
           
             7 
             24 
             DNA 
             Artificial Sequence 
             
               Description of Artificial Sequence
      Syntheticoligonucleotide 
             
           
            7
catagcagga acatcaacat ccac                                            24 
           
             8 
             1786 
             PRT 
             Artificial Sequence 
             
               Description of Artificial SequencePolypeptide 
             
           
            8
Met Thr Asn Ser Asn Tyr Lys Ser Asn Asn Lys Thr Tyr Asn Glu Asn
  1               5                  10                  15
Asn Asn Glu Gln Ile Thr Thr Ile Phe Asn Arg Thr Asn Met Asn Pro
             20                  25                  30
Ile Lys Lys Cys His Met Arg Glu Lys Ile Asn Lys Tyr Phe Phe Leu
         35                  40                  45
Ile Lys Ile Leu Thr Cys Thr Ile Leu Ile Trp Ala Val Gln Tyr Asp
     50                  55                  60
Asn Asn Ser Asp Ile Asn Lys Ser Trp Lys Lys Asn Thr Tyr Val Asp
 65                  70                  75                  80
Lys Lys Leu Asn Lys Leu Phe Asn Arg Ser Leu Gly Glu Ser Gln Val
                 85                  90                  95
Asn Gly Glu Leu Ala Ser Glu Glu Val Lys Glu Lys Ile Leu Asp Leu
            100                 105                 110
Leu Glu Glu Gly Asn Thr Leu Thr Glu Ser Val Asp Asp Asn Lys Asn
        115                 120                 125
Leu Glu Glu Ala Glu Asp Ile Lys Glu Asn Ile Leu Leu Ser Asn Ile
    130                 135                 140
Glu Glu Pro Lys Glu Asn Ile Ile Asp Asn Leu Leu Asn Asn Ile Gly
145                 150                 155                 160
Gln Asn Ser Glu Lys Gln Glu Ser Val Ser Glu Asn Val Gln Val Ser
                165                 170                 175
Asp Glu Leu Phe Asn Glu Leu Leu Asn Ser Val Asp Val Asn Gly Glu
            180                 185                 190
Val Lys Glu Asn Ile Leu Glu Glu Ser Gln Val Asn Asp Asp Ile Phe
        195                 200                 205
Asn Ser Leu Val Lys Ser Val Gln Gln Glu Gln Gln His Asn Val Glu
    210                 215                 220
Glu Lys Val Glu Glu Ser Val Glu Glu Asn Asp Glu Glu Ser Val Glu
225                 230                 235                 240
Glu Asn Val Glu Glu Asn Val Glu Glu Asn Asp Asp Gly Ser Val Ala
                245                 250                 255
Ser Ser Val Glu Glu Ser Ile Ala Ser Ser Val Asp Glu Ser Ile Asp
            260                 265                 270
Ser Ser Ile Glu Glu Asn Val Ala Pro Thr Val Glu Glu Ile Val Ala
        275                 280                 285
Pro Ser Val Val Glu Ser Val Ala Pro Ser Val Glu Glu Ser Val Glu
    290                 295                 300
Glu Asn Val Glu Glu Ser Val Ala Glu Asn Val Glu Glu Ser Val Ala
305                 310                 315                 320
Glu Asn Val Glu Glu Ser Val Ala Glu Asn Val Glu Glu Ser Val Ala
                325                 330                 335
Glu Asn Val Glu Glu Ile Val Ala Pro Thr Val Glu Glu Ile Val Ala
            340                 345                 350
Pro Thr Val Glu Glu Ile Val Ala Pro Ser Val Val Glu Ser Val Ala
        355                 360                 365
Pro Ser Val Glu Glu Ser Val Glu Glu Asn Val Glu Glu Ser Val Ala
    370                 375                 380
Glu Asn Val Glu Glu Ser Val Ala Glu Asn Val Glu Glu Ser Val Ala
385                 390                 395                 400
Glu Asn Val Glu Glu Ser Val Ala Glu Asn Val Glu Glu Ser Val Ala
                405                 410                 415
Glu Asn Val Glu Glu Ile Val Ala Pro Thr Val Glu Glu Ile Val Ala
            420                 425                 430
Pro Thr Val Glu Glu Ile Val Ala Pro Ser Val Val Glu Ser Val Ala
        435                 440                 445
Pro Ser Val Glu Glu Ser Val Glu Glu Asn Val Glu Glu Ser Val Ala
    450                 455                 460
Glu Asn Val Glu Glu Ser Val Ala Glu Asn Val Glu Glu Ser Val Ala
465                 470                 475                 480
Glu Asn Val Glu Glu Ser Val Ala Glu Asn Val Glu Glu Ser Val Ala
                485                 490                 495
Glu Asn Val Glu Glu Ser Val Ala Glu Asn Val Glu Glu Ser Val Ala
            500                 505                 510
Glu Asn Val Glu Glu Ile Val Ala Pro Thr Val Glu Glu Ile Val Ala
        515                 520                 525
Pro Thr Val Glu Glu Ile Val Ala Pro Ser Val Val Glu Ser Val Ala
    530                 535                 540
Pro Ser Val Glu Glu Ser Val Glu Glu Asn Val Glu Glu Ser Val Ala
545                 550                 555                 560
Glu Asn Val Glu Glu Ser Val Ala Glu Asn Val Glu Glu Ser Val Ala
                565                 570                 575
Glu Asn Val Glu Glu Ser Val Ala Glu Asn Val Glu Glu Ile Val Ala
            580                 585                 590
Pro Thr Val Glu Glu Ile Val Ala Pro Thr Val Glu Glu Ile Val Ala
        595                 600                 605
Pro Ser Val Val Glu Ser Val Ala Pro Ser Val Glu Glu Ser Val Glu
    610                 615                 620
Glu Asn Val Glu Glu Ser Val Ala Glu Asn Val Glu Glu Ser Val Ala
625                 630                 635                 640
Glu Asn Val Glu Glu Ser Val Ala Glu Asn Val Glu Glu Ile Val Ala
                645                 650                 655
Pro Thr Val Glu Glu Ile Val Ala Pro Thr Val Glu Glu Ile Val Ala
            660                 665                 670
Pro Ser Val Val Glu Ser Val Ala Pro Ser Val Glu Glu Ser Val Glu
        675                 680                 685
Glu Asn Val Glu Glu Ser Val Ala Glu Asn Val Glu Glu Ser Val Ala
    690                 695                 700
Glu Asn Val Glu Glu Ser Val Ala Glu Asn Val Glu Glu Ser Val Ala
705                 710                 715                 720
Glu Asn Val Glu Glu Ile Val Ala Pro Thr Val Glu Glu Ile Val Ala
                725                 730                 735
Pro Thr Val Glu Glu Ile Val Ala Pro Ser Val Val Glu Ser Val Ala
            740                 745                 750
Pro Ser Val Glu Glu Ser Val Glu Glu Asn Val Glu Glu Ser Val Ala
        755                 760                 765
Glu Asn Val Glu Glu Ser Val Ala Glu Asn Val Glu Glu Ser Val Ala
    770                 775                 780
Glu Asn Val Glu Glu Ser Val Ala Pro Thr Val Glu Glu Ile Val Ala
785                 790                 795                 800
Pro Ser Val Glu Glu Ser Val Ala Pro Ser Val Glu Glu Ser Val Ala
                805                 810                 815
Glu Asn Val Ala Thr Asn Leu Ser Asp Asn Leu Leu Ser Asn Leu Leu
            820                 825                 830
Gly Gly Ile Glu Thr Glu Glu Ile Lys Asp Ser Ile Leu Asn Glu Ile
        835                 840                 845
Glu Glu Val Lys Glu Asn Val Val Thr Thr Ile Leu Glu Asn Val Glu
    850                 855                 860
Glu Thr Thr Ala Glu Ser Val Thr Thr Phe Ser Asn Ile Leu Glu Glu
865                 870                 875                 880
Ile Gln Glu Asn Thr Ile Thr Asn Asp Thr Ile Glu Glu Lys Leu Glu
                885                 890                 895
Glu Leu His Glu Asn Val Leu Ser Ala Ala Leu Glu Asn Thr Gln Ser
            900                 905                 910
Glu Glu Glu Lys Lys Glu Val Ile Asp Val Ile Glu Glu Val Lys Glu
        915                 920                 925
Glu Val Ala Thr Thr Leu Ile Glu Thr Val Glu Gln Ala Glu Glu Lys
    930                 935                 940
Ser Ala Asn Thr Ile Thr Glu Ile Phe Glu Asn Leu Glu Glu Asn Ala
945                 950                 955                 960
Val Glu Ser Asn Glu Asn Val Ala Glu Asn Leu Glu Lys Leu Asn Glu
                965                 970                 975
Thr Val Phe Asn Thr Val Leu Asp Lys Val Glu Glu Thr Val Glu Ile
            980                 985                 990
Ser Gly Glu Ser Leu Glu Asn Asn Glu Met Asp Lys Ala Phe Phe Ser
        995                1000                1005
Glu Ile Phe Asp Asn Val Lys Gly Ile Gln Glu Asn Leu Leu Thr Gly
   1010                1015                1020
Met Phe Arg Ser Ile Glu Thr Ser Ile Val Ile Gln Ser Glu Glu Lys
1025               1030                1035                1040
Val Asp Leu Asn Glu Asn Val Val Ser Ser Ile Leu Asp Asn Ile Glu
               1045                1050                1055
Asn Met Lys Glu Gly Leu Leu Asn Lys Leu Glu Asn Ile Ser Ser Thr
           1060                1065                1070
Glu Gly Val Gln Glu Thr Val Thr Glu His Val Glu Gln Asn Val Tyr
       1075                1080                1085
Val Asp Val Asp Val Pro Ala Met Lys Asp Gln Phe Leu Gly Ile Leu
   1090                1095                1100
Asn Glu Ala Gly Gly Leu Lys Glu Met Phe Phe Asn Leu Glu Asp Val
1105               1110                1115                1120
Phe Lys Ser Glu Ser Asp Val Ile Thr Val Glu Glu Ile Lys Asp Glu
               1125                1130                1135
Pro Val Gln Lys Glu Val Glu Lys Glu Thr Val Ser Ile Ile Glu Glu
           1140                1145                1150
Met Glu Glu Asn Ile Val Asp Val Leu Glu Glu Glu Lys Glu Asp Leu
       1155                1160                1165
Thr Asp Lys Met Ile Asp Ala Val Glu Glu Ser Ile Glu Ile Ser Ser
   1170                1175                1180
Asp Ser Lys Glu Glu Thr Glu Ser Ile Lys Asp Lys Glu Lys Asp Val
1185               1190                1195                1200
Ser Leu Val Val Glu Glu Val Gln Asp Asn Asp Met Asp Glu Ser Val
               1205                1210                1215
Glu Lys Val Leu Glu Leu Lys Asn Met Glu Glu Glu Leu Met Lys Asp
           1220                1225                1230
Ala Val Glu Ile Asn Asp Ile Thr Ser Lys Leu Ile Glu Glu Thr Gln
       1235                1240                1245
Glu Leu Asn Glu Val Glu Ala Asp Leu Ile Lys Asp Met Glu Lys Leu
   1250                1255                1260
Lys Glu Leu Glu Lys Ala Leu Ser Glu Asp Ser Lys Glu Ile Ile Asp
1265               1270                1275                1280
Ala Lys Asp Asp Thr Leu Glu Lys Val Ile Glu Glu Glu His Asp Ile
               1285                1290                1295
Thr Thr Thr Leu Asp Glu Val Val Glu Leu Lys Asp Val Glu Glu Asp
           1300                1305                1310
Lys Ile Glu Lys Val Ser Asp Leu Lys Asp Leu Glu Glu Asp Ile Leu
       1315                1320                1325
Lys Glu Val Lys Glu Ile Lys Glu Leu Glu Ser Glu Ile Leu Glu Asp
   1330                1335                1340
Tyr Lys Glu Leu Lys Thr Ile Glu Thr Asp Ile Leu Glu Glu Lys Lys
1345               1350                1355                1360
Glu Ile Glu Lys Asp His Phe Glu Lys Phe Glu Glu Glu Ala Glu Glu
               1365                1370                1375
Ile Lys Asp Leu Glu Ala Asp Ile Leu Lys Glu Val Ser Ser Leu Glu
           1380                1385                1390
Val Glu Glu Glu Lys Lys Leu Glu Glu Val His Glu Leu Lys Glu Glu
       1395                1400                1405
Val Glu His Ile Ile Ser Gly Asp Ala His Ile Lys Gly Leu Glu Glu
   1410                1415                1420
Asp Asp Leu Glu Glu Val Asp Asp Leu Lys Gly Ser Ile Leu Asp Met
1425               1430                1435                1440
Leu Lys Gly Asp Met Glu Leu Gly Asp Met Asp Lys Glu Ser Leu Glu
               1445                1450                1455
Asp Val Thr Thr Lys Leu Gly Glu Arg Val Glu Ser Leu Lys Asp Val
           1460                1465                1470
Leu Ser Ser Ala Leu Gly Met Asp Glu Glu Gln Met Lys Thr Arg Lys
       1475                1480                1485
Lys Ala Gln Arg Pro Lys Leu Glu Glu Val Leu Leu Lys Glu Glu Val
   1490                1495                1500
Lys Glu Glu Pro Lys Lys Lys Ile Thr Lys Lys Lys Val Arg Phe Asp
1505               1510                1515                1520
Ile Lys Asp Lys Glu Pro Lys Asp Glu Ile Val Glu Val Glu Met Lys
               1525                1530                1535
Asp Glu Asp Ile Glu Glu Asp Val Glu Glu Asp Ile Glu Glu Asp Ile
           1540                1545                1550
Glu Glu Asp Lys Val Glu Asp Ile Asp Glu Asp Ile Asp Glu Asp Ile
       1555                1560                1565
Gly Glu Asp Lys Asp Glu Val Ile Asp Leu Ile Val Gln Lys Glu Lys
   1570                1575                1580
Arg Ile Glu Lys Val Lys Ala Lys Lys Lys Lys Leu Glu Lys Lys Val
1585               1590                1595                1600
Glu Glu Gly Val Ser Gly Leu Lys Lys His Val Asp Glu Val Met Lys
               1605                1610                1615
Tyr Val Gln Lys Ile Asp Lys Glu Val Asp Lys Glu Val Ser Lys Ala
           1620                1625                1630
Leu Glu Ser Lys Asn Asp Val Thr Asn Val Leu Lys Gln Asn Gln Asp
       1635                1640                1645
Phe Phe Ser Lys Val Lys Asn Phe Val Lys Lys Tyr Lys Val Phe Ala
   1650                1655                1660
Ala Pro Phe Ile Ser Ala Val Ala Ala Phe Ala Ser Tyr Val Val Gly
1665               1670                1675                1680
Phe Phe Thr Phe Ser Leu Phe Ser Ser Cys Val Thr Ile Ala Ser Ser
               1685                1690                1695
Thr Tyr Leu Leu Ser Lys Val Asp Lys Thr Ile Asn Lys Asn Lys Glu
           1700                1705                1710
Arg Pro Phe Tyr Ser Phe Val Phe Asp Ile Phe Lys Asn Leu Lys His
       1715                1720                1725
Tyr Leu Gln Gln Met Lys Glu Lys Phe Ser Lys Glu Lys Asn Asn Asn
   1730                1735                1740
Val Ile Glu Val Thr Asn Lys Ala Glu Lys Lys Gly Asn Val Gln Val
1745               1750                1755                1760
Thr Asn Lys Thr Glu Lys Thr Thr Lys Val Asp Lys Asn Asn Lys Val
               1765                1770                1775
Pro Lys Lys Arg Arg Thr Gln Lys Ser Lys
           1780                1785 
           
             9 
             630 
             PRT 
             Artificial Sequence 
             
               Description of Artificial SequencePolypeptide 
             
           
            9
Thr Leu Thr Glu Ser Val Asp Asp Asn Lys Asn Leu Glu Glu Ala Glu
  1               5                  10                  15
Asp Ile Lys Glu Asn Ile Leu Leu Ser Asn Ile Glu Glu Pro Lys Glu
             20                  25                  30
Asn Ile Ile Asp Asn Leu Leu Asn Asn Ile Gly Gln Asn Ser Glu Lys
         35                  40                  45
Gln Glu Ser Val Ser Glu Asn Val Gln Val Ser Asp Glu Leu Phe Asn
     50                  55                  60
Glu Leu Leu Asn Ser Val Asp Val Asn Gly Glu Val Lys Glu Asn Ile
 65                  70                  75                  80
Leu Glu Glu Ser Gln Val Asn Asp Asp Ile Phe Asn Ser Leu Val Lys
                 85                  90                  95
Ser Val Gln Gln Glu Gln Gln His Asn Val Glu Glu Lys Val Glu Glu
            100                 105                 110
Ser Val Glu Glu Asn Asp Glu Glu Ser Val Glu Glu Asn Val Glu Glu
        115                 120                 125
Asn Val Glu Glu Asn Asp Asp Gly Ser Val Ala Ser Ser Val Glu Glu
    130                 135                 140
Ser Ile Ala Ser Ser Val Asp Glu Ser Ile Asp Ser Ser Ile Glu Glu
145                 150                 155                 160
Asn Val Ala Pro Thr Val Glu Glu Ile Val Ala Pro Thr Val Glu Glu
                165                 170                 175
Ile Val Ala Pro Ser Val Val Glu Ser Val Ala Pro Ser Val Glu Glu
            180                 185                 190
Ser Val Ala Pro Ser Val Glu Glu Ser Val Ala Glu Asn Val Glu Glu
        195                 200                 205
Ser Val Ala Glu Asn Val Glu Glu Ile Val Ala Pro Ser Val Glu Glu
    210                 215                 220
Ser Val Ala Glu Asn Val Glu Glu Ser Val Ala Glu Asn Val Glu Glu
225                 230                 235                 240
Ser Val Ala Glu Asn Val Glu Glu Ser Val Ala Glu Asn Val Glu Glu
                245                 250                 255
Ser Val Ala Glu Asn Val Glu Glu Ile Val Ala Pro Thr Val Glu Glu
            260                 265                 270
Ser Val Ala Pro Thr Val Glu Glu Ile Val Ala Pro Thr Val Glu Glu
        275                 280                 285
Ser Val Ala Pro Thr Val Glu Glu Ile Val Val Pro Ser Val Glu Glu
    290                 295                 300
Ser Val Ala Pro Ser Val Glu Glu Ser Val Ala Glu Asn Val Glu Glu
305                 310                 315                 320
Ser Val Ala Glu Asn Val Glu Glu Ser Val Ala Glu Asn Val Glu Glu
                325                 330                 335
Ser Val Ala Glu Asn Val Glu Glu Ser Val Ala Glu Asn Val Glu Glu
            340                 345                 350
Ile Val Ala Pro Ser Val Glu Glu Ile Val Ala Pro Thr Val Glu Glu
        355                 360                 365
Ser Val Ala Glu Asn Val Ala Thr Asn Leu Ser Asp Asn Leu Leu Ser
    370                 375                 380
Asn Leu Leu Gly Gly Ile Glu Thr Glu Glu Ile Lys Asp Ser Ile Leu
385                 390                 395                 400
Asn Glu Ile Glu Glu Val Lys Glu Asn Val Val Thr Thr Ile Leu Glu
                405                 410                 415
Lys Val Glu Glu Thr Thr Ala Glu Ser Val Thr Thr Phe Ser Asn Ile
            420                 425                 430
Leu Glu Glu Ile Gln Glu Asn Thr Ile Thr Asn Asp Thr Ile Glu Glu
        435                 440                 445
Lys Leu Glu Glu Leu His Glu Asn Val Leu Ser Ala Ala Leu Glu Asn
    450                 455                 460
Thr Gln Ser Glu Glu Glu Lys Lys Glu Val Ile Asp Val Ile Glu Glu
465                 470                 475                 480
Val Lys Glu Glu Val Ala Thr Thr Leu Ile Glu Thr Val Glu Gln Ala
                485                 490                 495
Glu Glu Glu Ser Glu Ser Thr Ile Thr Glu Ile Phe Glu Asn Leu Glu
            500                 505                 510
Glu Asn Ala Val Glu Ser Asn Glu Lys Val Ala Glu Asn Leu Glu Lys
        515                 520                 525
Leu Asn Glu Thr Val Phe Asn Thr Val Leu Asp Lys Val Glu Glu Thr
    530                 535                 540
Val Glu Ile Ser Gly Glu Ser Leu Glu Asn Asn Glu Met Asp Lys Ala
545                 550                 555                 560
Phe Phe Ser Glu Ile Phe Asp Asn Val Lys Gly Ile Gln Glu Asn Leu
                565                 570                 575
Leu Thr Gly Met Phe Arg Ser Ile Glu Thr Ser Ile Val Ile Gln Ser
            580                 585                 590
Glu Glu Lys Val Asp Leu Asn Glu Asn Val Val Ser Ser Ile Leu Asp
        595                 600                 605
Asn Ile Glu Asn Met Lys Glu Gly Leu Leu Asn Lys Leu Glu Asn Ile
    610                 615                 620
Ser Ser Thr Glu Gly Glu
625                 630 
           
             10 
             50 
             PRT 
             Artificial Sequence 
             
               Description of Artificial SequencePolypeptide 
             
           
            10
Arg Asp Glu Leu Phe Asn Glu Leu Leu Asn Ser Val Asp Val Asn Gly
  1               5                  10                  15
Glu Val Lys Glu Asn Ile Leu Glu Glu Ser Gln Val Asn Asp Asp Ile
             20                  25                  30
Phe Asn Ser Leu Val Lys Ser Val Gln Gln Glu Gln Gln His Asn Val
         35                  40                  45
Glu Glu
     50 
           
             11 
             100 
             PRT 
             Artificial Sequence 
             
               Description of Artificial SequencePolypeptide 
             
           
            11
Val Glu Glu Ser Val Glu Glu Asn Asp Glu Glu Ser Val Glu Glu Asn
  1               5                  10                  15
Val Glu Glu Asn Val Glu Asn Asn Asp Asp Gly Ser Val Ala Ser Ser
             20                  25                  30
Val Glu Glu Ser Ile Ala Ser Ser Val Asp Glu Ser Ile Asp Ser Ser
         35                  40                  45
Ile Glu Glu Asn Val Ala Pro Thr Val Glu Glu Ile Val Ala Pro Thr
     50                  55                  60
Val Glu Glu Ile Val Ala Pro Ser Val Val Glu Lys Cys Ala Pro Ser
 65                  70                  75                  80
Val Glu Glu Ser Val Ala Pro Ser Val Glu Glu Ser Val Ala Glu Met
                 85                  90                  95
Leu Lys Glu Arg
            100 
           
             12 
             47 
             PRT 
             Artificial Sequence 
             
               Description of Artificial SequencePolypeptide 
             
           
            12
Arg Asp Glu Leu Phe Asn Glu Leu Leu Asn Ser Val Asp Val Asn Gly
  1               5                  10                  15
Glu Val Lys Glu Asn Ile Leu Glu Glu Ser Gln Val Asn Asp Asp Ile
             20                  25                  30
Phe Asn Ser Leu Val Lys Ser Val Gln Gln Glu Gln Gln His Asn
         35                  40                  45 
           
             13 
             26 
             PRT 
             Artificial Sequence 
             
               Description of Artificial SequencePolypeptide 
             
           
            13
Asp Glu Leu Phe Asn Glu Leu Leu Asn Ser Val Asp Val Asn Gly Glu
  1               5                  10                  15
Val Lys Glu Asn Ile Leu Glu Glu Ser Gln
             20                  25 
           
             14 
             27 
             PRT 
             Artificial Sequence 
             
               Description of Artificial SequencePolypeptide 
             
           
            14
Leu Glu Glu Ser Gln Val Asn Asp Asp Ile Phe Ser Asn Ser Leu Val
  1               5                  10                  15
Lys Ser Val Gln Gln Glu Gln Gln His Asn Val
             20                  25 
           
             15 
             28 
             PRT 
             Artificial Sequence 
             
               Description of Artificial SequencePolypeptide 
             
           
            15
Val Glu Ser Val Ala Pro Ser Val Glu Glu Ser Val Ala Pro Ser Val
  1               5                  10                  15
Glu Glu Ser Val Ala Glu Asn Val Glu Glu Ser Val
             20                  25 
           
             16 
             20 
             PRT 
             Artificial Sequence 
             
               Description of Artificial SequencePolypeptide 
             
           
            16
Leu Leu Ser Asn Ile Glu Glu Pro Lys Glu Asn Ile Ile Asp Asn Leu
  1               5                  10                  15
Leu Asn Asn Ile
             20 
           
             17 
             4 
             PRT 
             Artificial Sequence 
             
               Description of Artificial SequencePolypeptide 
             
           
            17
Val Glu Glu Ser
  1 
           
             18 
             4 
             PRT 
             Artificial Sequence 
             
               Description of Artificial SequencePolypeptide 
             
           
            18
Val Glu Glu Asn
  1 
           
             19 
             4 
             PRT 
             Artificial Sequence 
             
               Description of Artificial SequencePolypeptide 
             
           
            19
Val Glu Glu Ile
  1 
           
             20 
             4 
             PRT 
             Artificial Sequence 
             
               Description of Artificial SequencePolypeptide 
             
           
            20
Val Ala Pro Ser
  1 
           
             21 
             56 
             PRT 
             Artificial Sequence 
             
               Description of Artificial SequencePolypeptide 
             
           
            21
Val Glu Glu Lys Val Glu Glu Ser Val Glu Glu Asn Asp Glu Glu Ser
  1               5                  10                  15
Val Glu Glu Asn Val Glu Glu Asn Val Glu Glu Asn Asp Asp Gly Ser
             20                  25                  30
Val Ala Ser Ser Val Glu Glu Ser Ile Ala Ser Ser Val Asp Glu Ser
         35                  40                  45
Ile Asp Ser Ser Ile Glu Glu Asn
     50                  55 
           
             22 
             540 
             PRT 
             Artificial Sequence 
             
               Description of Artificial SequencePolypeptide 
             
           
            22
Val Ala Pro Thr Val Glu Glu Ile Val Ala Pro Ser Val Val Glu Ser
  1               5                  10                  15
Val Ala Pro Ser Val Glu Glu Ser Val Glu Glu Asn Val Glu Glu Ser
             20                  25                  30
Val Ala Glu Asn Val Glu Glu Ser Val Ala Glu Asn Val Glu Glu Ser
         35                  40                  45
Val Ala Glu Asn Val Glu Glu Ser Val Ala Glu Asn Val Glu Glu Ile
     50                  55                  60
Val Ala Pro Thr Val Glu Glu Ile Val Ala Pro Thr Val Glu Glu Ile
 65                  70                  75                  80
Val Ala Pro Ser Val Val Glu Ser Val Ala Pro Ser Val Glu Glu Ser
                 85                  90                  95
Val Glu Glu Asn Val Glu Glu Ser Val Ala Glu Asn Val Glu Glu Ser
            100                 105                 110
Val Ala Glu Asn Val Glu Glu Ser Val Ala Glu Asn Val Glu Glu Ser
        115                 120                 125
Val Ala Glu Asn Val Glu Glu Ser Val Ala Glu Asn Val Glu Glu Ile
    130                 135                 140
Val Ala Pro Thr Val Glu Glu Ile Val Ala Pro Thr Val Glu Glu Ile
145                 150                 155                 160
Val Ala Pro Ser Val Val Glu Ser Val Ala Pro Ser Val Glu Glu Ser
                165                 170                 175
Val Glu Glu Asn Val Glu Glu Ser Val Ala Glu Asn Val Glu Glu Ser
            180                 185                 190
Val Ala Glu Asn Val Glu Glu Ser Val Ala Glu Asn Val Glu Glu Ser
        195                 200                 205
Val Ala Glu Asn Val Glu Glu Ser Val Ala Glu Asn Val Glu Glu Ser
    210                 215                 220
Val Ala Glu Asn Val Glu Glu Ser Val Ala Glu Asn Val Glu Glu Ile
225                 230                 235                 240
Val Ala Pro Thr Val Glu Glu Ile Val Ala Pro Thr Val Glu Glu Ile
                245                 250                 255
Val Ala Pro Ser Val Val Glu Ser Val Ala Pro Ser Val Glu Glu Ser
            260                 265                 270
Val Glu Glu Asn Val Glu Glu Ser Val Ala Glu Asn Val Glu Glu Ser
        275                 280                 285
Val Ala Glu Asn Val Glu Glu Ser Val Ala Glu Asn Val Glu Glu Ser
    290                 295                 300
Val Ala Glu Asn Val Glu Glu Ile Val Ala Pro Thr Val Glu Glu Ile
305                 310                 315                 320
Val Ala Pro Thr Val Glu Glu Ile Val Ala Pro Ser Val Val Glu Ser
                325                 330                 335
Val Ala Pro Ser Val Glu Glu Ser Val Glu Glu Asn Val Glu Glu Ser
            340                 345                 350
Val Ala Glu Asn Val Glu Glu Ser Val Ala Glu Asn Val Glu Glu Ser
        355                 360                 365
Val Ala Glu Asn Val Glu Glu Ile Val Ala Pro Thr Val Glu Glu Ile
    370                 375                 380
Val Ala Pro Thr Val Glu Glu Ile Val Ala Pro Ser Val Val Glu Ser
385                 390                 395                 400
Val Ala Pro Ser Val Glu Glu Ser Val Glu Glu Asn Val Glu Glu Ser
                405                 410                 415
Val Ala Glu Asn Val Glu Glu Ser Val Ala Glu Asn Val Glu Glu Ser
            420                 425                 430
Val Ala Glu Asn Val Glu Glu Ser Val Ala Glu Asn Val Glu Glu Ile
        435                 440                 445
Val Ala Pro Thr Val Glu Glu Ile Val Ala Pro Thr Val Glu Glu Ile
    450                 455                 460
Val Ala Pro Ser Val Val Glu Ser Val Ala Pro Ser Val Glu Glu Ser
465                 470                 475                 480
Val Glu Glu Asn Val Glu Glu Ser Val Ala Glu Asn Val Glu Glu Ser
                485                 490                 495
Val Ala Glu Asn Val Glu Glu Ser Val Ala Glu Asn Val Glu Glu Ser
            500                 505                 510
Val Ala Pro Thr Val Glu Glu Ile Val Ala Pro Ser Val Glu Glu Ser
        515                 520                 525
Val Ala Pro Ser Val Glu Glu Ser Val Ala Glu Asn
    530                 535                 540 
           
             23 
             39 
             PRT 
             Artificial Sequence 
             
               Description of Artificial SequencePolypeptide 
             
           
            23
Asp Glu Asp Ile Glu Glu Asp Val Glu Glu Asp Ile Glu Glu Asp Ile
  1               5                  10                  15
Glu Glu Asp Lys Val Glu Asp Ile Asp Glu Asp Ile Asp Glu Asp Ile
             20                  25                  30
Gly Glu Asp Lys Asp Glu Val
         35 
           
             24 
             56 
             PRT 
             Artificial Sequence 
             
               Description of Artificial SequencePolypeptide 
             
           
            24
Val Glu Glu Lys Val Glu Glu Ser Val Glu Glu Asn Asp Glu Glu Ser
  1               5                  10                  15
Val Glu Glu Asn Val Glu Glu Asn Val Glu Glu Asn Asp Asp Gly Ser
             20                  25                  30
Val Ala Ser Ser Val Glu Glu Ser Ile Ala Ser Ser Val Asp Glu Ser
         35                  40                  45
Ile Asp Ser Ser Ile Glu Glu Asn
     50                  55 
           
             25 
             212 
             PRT 
             Artificial Sequence 
             
               Description of Artificial SequencePolypeptide 
             
           
            25
Val Ala Pro Thr Val Glu Glu Ile Val Ala Pro Thr Val Glu Glu Ile
  1               5                  10                  15
Val Ala Pro Ser Val Val Glu Ser Val Ala Pro Ser Val Glu Glu Ser
             20                  25                  30
Val Ala Pro Ser Val Glu Glu Ser Val Ala Glu Asn Val Glu Glu Ser
         35                  40                  45
Val Ala Glu Asn Val Glu Glu Ile Val Ala Pro Ser Val Glu Glu Ser
     50                  55                  60
Val Ala Glu Asn Val Glu Glu Ser Val Ala Glu Asn Val Glu Glu Ser
 65                  70                  75                  80
Val Ala Glu Asn Val Glu Glu Ser Val Ala Glu Asn Val Glu Glu Ser
                 85                  90                  95
Val Ala Glu Asn Val Glu Glu Ile Val Ala Pro Thr Val Glu Glu Ser
            100                 105                 110
Val Ala Pro Thr Val Glu Glu Ile Val Ala Pro Thr Val Glu Glu Ser
        115                 120                 125
Val Ala Pro Thr Val Glu Glu Ile Val Val Pro Ser Val Glu Glu Ser
    130                 135                 140
Val Ala Pro Ser Val Glu Glu Ser Val Ala Glu Asn Val Glu Glu Ser
145                 150                 155                 160
Val Ala Glu Asn Val Glu Glu Ser Val Ala Glu Asn Val Glu Glu Ser
                165                 170                 175
Val Ala Glu Asn Val Glu Glu Ser Val Ala Glu Asn Val Glu Glu Ile
            180                 185                 190
Val Ala Pro Ser Val Glu Glu Ile Val Ala Pro Thr Val Glu Glu Ser
        195                 200                 205
Val Ala Glu Asn
    210 
           
             26 
             4 
             PRT 
             Artificial Sequence 
             
               Description of Artificial SequencePolypeptide 
             
           
            26
Val Val Glu Ser
  1 
           
             27 
             4 
             PRT 
             Artificial Sequence 
             
               Description of Artificial SequencePolypeptide 
             
           
            27
Val Ala Glu Asn
  1 
           
             28 
             4 
             PRT 
             Artificial Sequence 
             
               Description of Artificial SequencePolypeptide 
             
           
            28
Val Ala Pro Thr 
           
             29 
             4 
             PRT 
             Artificial Sequence 
             
               Description of Artificial SequencePolypeptide 
             
           
            29
Val Val Pro Ser