Abstract:
The present invention relates to antigens, more particularly an antigen of  Streptococcus pygenes  (also called group A Streptococcus (GAS)) bacterial pathogen which is useful as vaccine component for therapy and/or prophylaxis.

Description:
FIELD OF THE INVENTION  
         [0001]    The present invention is related to antigens, more particularly a polypeptide antigen of  Streptococcus pyogenes  (also called group A Streptococcus (GAS)) bacterial pathogen which may be useful for prophylaxis, diagnostic and/or therapy of streptococcal infection.  
         BACKGROUND OF THE INVENTION  
         [0002]    Streptococci are gram (+) bacteria which are differentiated by group specific carbohydrate antigens A through O which are found at the cell surface.  Streptococcus pyogenes  isolates are further distinguished by type-specific M protein antigens. M proteins are important virulence factors which are highly variable both in molecular weights and in sequences. Indeed, more than 80-M protein types have been identified on the basis of antigenic differences.  
           [0003]    [0003] Streptococcus pyogenes  is responsible for many diverse infection types, including pharyngitis, erysipelas and impetigo, scarlet fever, and invasive diseases such as bacteremia and necrotizing fasciitis and also toxic shock. A resurgence of invasive disease in recent years has been documented in many countries, including those in North America and Europe. Although the organism is sensitive to antibiotics, the high attack rate and rapid onset of sepsis results in high morbidity and mortality.  
           [0004]    To develop a vaccine that will protect individuals from  Streptococcus pyogenes  infection, efforts have concentrated on virulence factors such as the type-specific M proteins. However, the amino-terminal portion of M proteins was found to induce cross-reactive antibodies which reacted with human myocardium, tropomyosin, myosin, and vimentin, which might be implicated in autoimmune diseases. Others have used recombinant techniques to produce complex hybrid proteins containing amino-terminal peptides of M proteins from different serotypes. However, a safe vaccine containing all  Streptococcus pyogenes  serotypes will be highly complex to produce and standardize.  
           [0005]    In addition to the serotype-specific antigens, other  Streptococcus pyogenes  proteins have generated interest as potential vaccine candidates. The C5a peptidase, which is expressed by at least  Streptococcus pyogenes  40 serotypes, was shown to be immunogenic in mice, but its capacity to reduce the level of nasopharyngeal colonization was limited. Other investigators have also focused on the streptococcal pyrogenic exotoxins which appear to play an important role in pathogenesis of infection. Immunization with these proteins prevented the deadly symptoms of toxic shock, but did not prevent colonization.  
           [0006]    Therefore there remains an unmet need for  Streptococcus pyogenes  antigens that may be used vaccine components for prophylaxis, diagnostic and/or therapy of Streptococcus infection.  
         SUMMARY OF THE INVENTION  
         [0007]    According to one aspect, the present invention provides an isolated polynucleotide encoding a polypeptide having at least 70% identity to a second polypeptide comprising a sequence chosen from SEQ ID NOs: 2, 4, 6, 8, 10, 12, 14, 16, 20 or fragments, analogues or derivatives thereof.  
           [0008]    According to one aspect, the present invention provides an isolated polynucleotide encoding a polypeptide having at least 95% identity to a second polypeptide comprising a sequence chosen from SEQ ID NOs: 2, 4, 6, 8, 10, 12, 14, 16, 20 or fragments, analogues or derivatives thereof.  
           [0009]    In other aspects, there are provided novel polypeptides encoded by polynucleotides of the invention, vectors comprising polynucleotides of the invention operably linked to an expression control region, as well as host cells transfected with said vectors, pharmaceutical or vaccine compositions and methods of producing polypeptides comprising culturing said host cells under conditions suitable for expression. 
       
    
    
     BRIEF DESCRIPTION OF THE DRAWINGS  
       [0010]    [0010]FIG. 1 is the DNA sequence of BVH-P1 gene from serotype 3  S. pyogenes  strain ATCC12384 with a secretion signal at position 1 to 75; SEQ ID NO:1.  
         [0011]    [0011]FIG. 2 is the amino acid sequence BVH-P1 protein from serotype 3  S. pyogenes  strain ATCC12384 with a secretion signal at position 1 to 25; SEQ ID NO:2.  
         [0012]    [0012]FIG. 3 is the DNA sequence of BVH-P1 gene from  S. pyogenes  strain LSPQ2699(ATCC19615) with a secretion signal at position 1 to 75; SEQ ID NO:3.  
         [0013]    [0013]FIG. 4 is the amino acid sequence BVH-P1 protein from  S. pyogenes  strain LSPQ2699(ATCC19615) with a secretion signal at position 1 to 25; SEQ ID NO:4.  
         [0014]    [0014]FIG. 5 is the DNA sequence of BVH-P1 gene from  S. pyogenes  strain SPY57 with a secretion signal at position 1 to 75; SEQ ID NO:5. FIG. 6 is the amino acid sequence BVH-P1 protein from  S. pyogenes  strain SPY57 with a secretion signal at position 1 to 25; SEQ ID NO:6.  
         [0015]    [0015]FIG. 7 is the DNA sequence of BVH-P1 gene from  S. pyogenes  strain B514 with a secretion signal at position 1 to 75; SEQ ID NO:7.  
         [0016]    [0016]FIG. 8 is the amino acid sequence BVH-P1 protein from  S. pyogenes  strain B514 with a secretion signal at position 1 to 25; SEQ ID NO:8.  
         [0017]    [0017]FIG. 9 is the DNA sequence BVH-P1 gene without a secretion signal from serotype 3  S. pyogenes  strain ATCC12384; SEQ ID NO:9.  
         [0018]    [0018]FIG. 10 is the amino acid sequence BVH-P1 protein without a secretion signal from serotype 3  S. pyogenes  strain ATCC12384 SEQ ID NO:10.  
         [0019]    [0019]FIG. 11 is the DNA sequence BVH-P1 gene without a secretion signal from serotype 3  S. pyogenes  strain LSPQ2699 (ATCC19615); SEQ ID NO:11.  
         [0020]    [0020]FIG. 12 is the amino acid sequence BVH-P1 protein without a secretion signal from serotype 3  S. pyogenes strain LSPQ 2699 (ATCC19615); SEQ ID NO:12.  
         [0021]    [0021]FIG. 13 is the DNA sequence BVH-P1 gene without a secretion signal from serotype 3  S. pyogenes  strain SPY57; SEQ ID NO:13.  
         [0022]    [0022]FIG. 14 is the amino acid sequence BVH-P1 protein without a secretion signal from serotype 3  S. pyogenes  strain SPY57; SEQ ID NO:14.  
         [0023]    [0023]FIG. 15 is the DNA sequence BVH-P1 gene without a secretion signal from serotype 3  S. pyogenes  strain B514; SEQ ID NO:15.  
         [0024]    [0024]FIG. 16 is the amino acid sequence BVH-P1 protein without a secretion signal from serotype 3  S. pyogenes  strain B514; SEQ ID NO:16.  
         [0025]    FIG. 17 depicts the comparison of the nucleotide sequences of the BVH-P1 genes from ATCC12384, LSPQ2699(ATCC19615), SPY57, B514, ATCC 70029 (Oklahoma) and T28/51/4 (UO9352)  S. pyogenes  strains by using the program Clustal W from MacVector sequence analysis software (version 6.5). Underneath the alignment, there is a consensus line. Shaded nucleotides are identical between every sequences and gaps in the sequence introduced by alignment are indicated by hyphens.  
         [0026]    FIG. 18 depicts the comparison of the predicted amino acid sequences of the BVH-P1 open reading frames from ATCC12384, LSPQ2699(ATCC19615), SPY57, B514, ATCC 70029 (Oklahoma) and T28/51/4 (UO9352)  S. pyogenes  strains by using the program Clustal W from MacVector sequence analysis software (version 6.5). Underneath the alignment, there is a consensus line. Shaded amino acid residues are identical between every sequences and gaps in the sequence introduced by alignment are indicated by hyphens.  
         [0027]    [0027]FIG. 19 is the DNA sequence of a gene from  S. pneumonia;  SEQ ID NO:17.  
         [0028]    [0028]FIG. 20 is the amino acid sequence of a protein from  S. pneumonia;  SEQ ID NO:18. 
     
    
     DETAILED DESCRIPTION OF THE INVENTION  
       [0029]    According to one aspect, the present invention provides an isolated polynucleotide encoding a polypeptide having at least 70% identity to a second polypeptide comprising a sequence chosen from SEQ ID NOs: 2, 4, 6, 8, 10, 12, 14, 16, 20 or fragments, analogues or derivatives thereof.  
         [0030]    According to one aspect, the present invention provides an isolated polynucleotide encoding a polypeptide having at least 95% identity to a second polypeptide comprising a sequence chosen from SEQ ID NOs: 2, 4, 6, 8, 10, 12, 14, 16, 20 or fragments, analogues or derivatives thereof.  
         [0031]    According to one aspect, the present invention relates to polypeptides characterized by the amino acid sequence comprising SEQ ID NOs: 2, 4, 6, 8, 10, 12, 14, 16, 20 or fragments, analogues or derivatives thereof.  
         [0032]    According to one aspect, the present invention provides an isolated polynucleotide encoding a polypeptide capable of generating antibodies having binding specificity for a polypeptide comprising a sequence chosen from SEQ ID NOs: 2, 4, 6, 8, 10, 12, 14, 16, 20 or fragments, analogues or derivatives thereof.  
         [0033]    In accordance with the present invention, there is provided a consensus nucleotide sequence depicted in FIG. 17. As can be seen by the alignement, the polynucleotide encoding the polypeptide of the invention is well conserved. Without restricting the scope of the invention, the following table 1 shows the possible modifications. SEQ ID NO:19 covers the consensus nucleotide sequence depicted in FIG. 17 with the modifications illustrated in Table 1: 
                                                   Position on alignement in                   Possible nucleotide                            21   C or T            53   C or T            69   G or A           103   G or C           149   C or T           150   A or T           195   G or A           244   T or C           273   A or C           282   T or C           302   C or A           318   A or G           334   G or T           394   C or T           400   G or A           415   C or T           428-448   [CTGATGTCCCAACGACACCAT] or               none           450   C or A           473   C or T           501   G or A           527   T or C           572   T or A           573   T or A           595   A or C           596   C or G           597   G or C           630   A or G           632   A or C           633   C or T           634   C or T           665   A or G           666   G or A           683   T or C           708   C or T           733   [CAGATGTTAACT] or none           798   T or C           883   G or none           927   T or A           930   T or C           943   T or none           952   T or A           955   G or A           964   T or C           973   G or A           976   T or G           978   A or T           979   A or T           981   A or G           982   T or C           986   G or A           988   T or G           1033    G or C           1034    C or G           1102    C or T           1143    A or T           1144    A or T           1145    A or T           1146    A or T                      
 
         [0034]    In accordance with the present invention, there is provided a consensus amino acid sequence depicted in FIG. 18. As can be seen by the alignement, the polypeptide of the invention is well conserved. Without restricting the scope of the invention, the following table 2 shows the possible modifications. SEQ ID NO:20 covers the consensus nucleotide sequence depicted in FIG. 18 with the modifications illustrated in Table 2: 
                                                   Position on alignement in                   Possible amino acid                            18   A or V            35   E or Q            50   T or I           101   T or N           112   A or S           132   P or S           134   V or I           139   S or P           143 to 149   SDVPTTP or none           150   F or L           158   S or P           176   L or S           191   V or E           199   T or P or S           211   D or A           212   P or S           222   E or G           228   V or A           242 to 245   ETSQ or none           246   E or M           247   T or L           248   S or T           295   A or L           296   S or L           297   A or P           298   F or L           299   G or V           300   I or L           301   T or R           302   S or H           303   F or L           304   S or V           305   G or V           306   Y or T           307   R or V           308   P or Q           309   G or E           310   D or I           311   P or Q           312   G or E           313   D or I           314   H or I           326   E or V           327   N or S           329   A or T           344   E or D           345   R or G           380   E or V           381   N or F                      
 
         [0035]    In accordance with the present invention, all polynucleotides encoding polypeptides are within the scope of the present invention.  
         [0036]    In a further embodiment, the polypeptides in accordance with the present invention are antigenic.  
         [0037]    In a further embodiment, the polypeptides in accordance with the present invention are immunogenic.  
         [0038]    In a further embodiment, the polypeptides in accordance with the present invention can elicit an immune response in an individual.  
         [0039]    In a further embodiment, the present invention also relates to polypeptides which are able to raise antibodies having binding specificity to the polypeptides of the present invention as defined above.  
         [0040]    An antibody that “has binding specificity” is an antibody that recognizes and binds the selected polypeptide but which does not substantially recognize and bind other molecules in a sample, e.g., a biological sample. Specific binding can be measured using an ELISA assay in which the selected polypeptide is used as an antigen.  
         [0041]    In accordance with the present invention, “protection” in the biological studies is defined by a significant increase in the survival curve, rate or period. Statistical analysis using the Log rank test to compare survival curves, and Fisher exact test to compare survival rates and numbers of days to death, respectively, might be useful to calculate P values and determine whether the difference between the two groups is statistically significant. P values of 0.05 are regarded as not significant.  
         [0042]    As used herein, “fragments”, “analogues” or “derivatives” of the polypeptides of the invention include those polypeptides in which one or more of the amino acid residues are substituted with a conserved or non-conserved amino acid residue (preferably conserved) and which may be natural or unnatural. In one embodiment, derivatives and analogues of polypeptides of the invention will have about 70% identity with those sequences illustrated in the figures or fragments thereof. That is, 70% of the residues are the same. In a further embodiment, polypeptides will have greater than 75% homology. In a further embodiment, polypeptides will have greater than 80% homology. In a further embodiment, polypeptides will have greater than 85% homology. In a further embodiment, polypeptides will have greater than 90% homology. In a further embodiment, polypeptides will have greater than 95% homology. In a further embodiment, polypeptides will have greater than 99% homology. In a further embodiment, derivatives and analogues of polypeptides of the invention will have less than about 20 amino acid residue substitutions, modifications or deletions and more preferably less than 10. Preferred substitutions are those known in the art as conserved i.e. the substituted residues share physical or chemical properties such as hydrophobicity, size, charge or functional groups.  
         [0043]    The skilled person will appreciate that fragments, analogues or derivatives of the proteins or polypeptides of the invention will also find use in the context of the present invention, i.e. as antigenic/immunogenic material. Thus, for instance proteins or polypeptides which include one or more additions, deletions, substitutions or the like are encompassed by the present invention. In addition, it may be possible to replace one amino acid with another of similar “type”. For instance replacing one hydrophobic amino acid with another hydropholic amino acid.  
         [0044]    One can use a program such as the CLUSTAL program to compare amino acid sequences. This program compares amino acid sequences and finds the optimal alignment by inserting spaces in either sequence as appropriate. It is possible to calculate amino acid identity or similarity (identity plus conservation of amino acid type) for an optimal alignment. A program like BLASTx will align the longest stretch of similar sequences and assign a value to the fit. It is thus possible to obtain a comparison where several regions of similarity are found, each having a different score. Both types of identity analysis are contemplated in the present invention.  
         [0045]    In an alternative approach, the analogues or derivatives could be fusion proteins, incorporating moieties which render purification easier, for example by effectively tagging the desired protein or polypeptide, it may be necessary to remove the “tag” or it may be the case that the fusion protein itself retains sufficient antigenicity to be useful.  
         [0046]    In an additional aspect of the invention there are provided antigenic/immunogenic fragments of the proteins or polypeptides of the invention, or of analogues or derivatives thereof.  
         [0047]    The fragments of the present invention should include one or more epitopic regions or be sufficiently similar to such regions to retain their antigenic/immunogenic properties. Thus, for fragments according to the present invention the degree of identity is perhaps irrelevant, since they may be 100% identical to a particular part of a protein or polypeptide, homologue or derivative as described herein. The key issue, once again, is that the fragment retains the antigenic/immunogenic properties.  
         [0048]    Thus, what is important for analogues, derivatives and fragments is that they possess at least a degree of the antigenicity/immunogenic of the protein or polypeptide from which they are derived.  
         [0049]    Also included are polypeptides which have fused thereto other compounds which alter the polypeptides biological or pharmacological properties i.e. polyethylene glycol (PEG) to increase half-life; leader or secretory amino acid sequences for ease of purification; prepro- and pro-sequences; and (poly)saccharides.  
         [0050]    Furthermore, in those situations where amino acid regions are found to be polymorphic, it may be desirable to vary one or more particular amino acids to more effectively mimic the different epitopes of the different streptococcus strains.  
         [0051]    Moreover, the polypeptides of the present invention can be modified by terminal —NH 2  acylation (eg. by acetylation, or thioglycolic acid amidation, terminal carbosy amidation, e.g. with ammonia or methylamine) to provide stability, increased hydrophobicity for linking or binding to a support or other molecule.  
         [0052]    Also contemplated are hetero and homo polypeptide multimers of the polypeptide fragments, analogues and derivatives. These polymeric forms include, for example, one or more polypeptides that have been cross-linked with cross-linkers such as avidin/biotin, gluteraldehyde or dimethylsuperimidate. Such polymeric forms also include polypeptides containing two or more tandem or inverted contiguous sequences, produced from multicistronic mRNAs generated by recombinant DNA technology.  
         [0053]    Preferably, a fragment, analog or derivative of a polypeptide of the invention will comprise at least one antigenic region i.e. at least one epitope.  
         [0054]    In order to achieve the formation of antigenic polymers (i.e. synthetic multimers), polypeptides may be utilized having bishaloacetyl groups, nitroarylhalides, or the like, where the reagents being specific for thio groups. Therefore, the link between two mercapto groups of the different peptides may be a single bond or may be composed of a linking group of at least two, typically at least four, and not more than 16, but usually not more than about 14 carbon atoms.  
         [0055]    In a particular embodiment, polypeptide fragments, analogues and derivatives of the invention do not contain a methionine (Met) starting residue. Preferably, polypeptides will not incorporate a leader or secretory sequence (signal sequence). The signal portion of a polypeptide of the invention may be determined according to established molecular biological techniques. In general, the polypeptide of interest may be isolated from a streptococcal culture and subsequently sequenced to determine the initial residue of the mature protein and therefore the sequence of the mature polypeptide.  
         [0056]    According to another aspect, there are provided vaccine compositions comprising one or more streptococcal polypeptides of the invention in admixture with a pharmaceutically acceptable carrier diluent or adjuvant. Suitable adjuvants include oils i.e. Freund&#39;s complete or incomplete adjuvant; salts i.e. AlK(SO 4 ) 2 , AlNa(SO 4 ) 2 , AlNH 4  (SO 4 ) 2 , silica, kaolin, carbon polynucleotides i.e. poly IC and poly AU. Preferred adjuvants include QuilA and Alhydrogel. Vaccines of the invention may be administered parenterally by injection, rapid infusion, nasopharyngeal absorption, dermoabsorption, or bucal or oral. Pharmaceutically acceptable carriers also include tetanus toxoid.  
         [0057]    The term vaccine is also meant to include antibodies. In accordance with the present invention, there is also provided the use of one or more antibodies having binding specificity for the polypeptides of the present invention for the treatment or prophylaxis of streptococcus infection and/or diseases and symptoms mediated by streptococcus infection.  
         [0058]    Vaccine compositions of the invention are used for the treatment or prophylaxis of streptococcal infection and/or diseases and symptoms mediated by streptococcal infection As described in P. R. Murray (Ed, in chief),E. J. Baron, M. A. Pfaller, F. C. Tenover and R. H. Yolken. Manual of Clinical Microbiology, ASM Press, Washington, D.C. sixth edition, 1995, 1482p which are herein incorporated by reference. In one embodiment, vaccine compositions of the present invention are used for the prophylaxis or treatment of pharyngitis, erysipelas and impetigo, scarlet fever, and invasive diseases such as bacteremia and necrotizing fasciitis and also toxic shock. In one embodiment, vaccine compositions of the invention are used for the prophylaxis or treatment of streptococcus infection and/or diseases and symptoms mediated by streptococcus infection, in particular group A streptococcus ( pyogenes ), group B streptococcus (GBS or agalactiae),  S. pneumoniae,  dysgalactiae, uberis, nocardia as well as  Staphylococcus aureus.  In a further embodiment, the streptococcus infection is  Streptococcus pyogenes.    
         [0059]    In a particular embodiment, vaccines are administered to those individuals at risk of streptococcus infection such as infants, elderly and immunocompromised individuals.  
         [0060]    As used in the present application, the term “individuals” include mammals. In a further embodiment, the mammal is human.  
         [0061]    Vaccine compositions are preferably in unit dosage form of about 0.001 to 100 μg/kg (antigen/body weight) and more preferably 0.01 to 10 μg/kg and most preferably 0.1 to 1 μg/kg 1 to 3 times with an interval of about 1 to 6 week intervals between immunizations.  
         [0062]    Vaccine compositions are preferably in unit dosage form of about 0.1 μg to 10 mg and more preferably 1 μg to 1 mg and most preferably 10 to 100 μg 1 to 3 times with an interval of about 1 to 6 week intervals between immunizations.  
         [0063]    According to another aspect, there are provided polynucleotides encoding polypeptides characterized by the amino acid sequence chosen from SEQ ID NOs: 2, 4, 6, 8, 10, 12, 14, 16, 20 or fragments, analogues or derivatives thereof.  
         [0064]    In one embodiment, polynucleotides are those illustrated in SEQ ID NOs: 1, 3, 5, 7, 9, 11, 13, 15, 19 which may include the open reading frames (ORF), encoding polypeptides of the invention.  
         [0065]    It will be appreciated that the polynucleotide sequences illustrated in the figures may be altered with degenerate codons yet still encode the polypeptides of the invention. Accordingly the present invention further provides polynucleotides which hybridize to the polynucleotide sequences herein above described (or the complement sequences thereof) having 50% identity between sequences. In one embodiment, at least 70% identity between sequences. In one embodiment, at least 75% identity between sequences. In one embodiment, at least 80% identity between sequences. In one embodiment, at least 85% identity between sequences. In one embodiment, at least 90% identity between sequences. In a further embodiment, polynucleotides are hybridizable under stringent conditions i.e. having at least 95% identity. In a further embodiment, more than 97% identity.  
         [0066]    Suitable stringent conditions for hybridation can be readily determined by one of skilled in the art (see for example Sambrook et al., (1989) Molecular cloning: A Laboratory Manual, 2 nd  ed, Cold Spring Harbor, N.Y.; Current Protocols in Molecular Biology, (1999) Edited by Ausubel F. M. et al., John Wiley &amp; Sons, Inc., N.Y.).  
         [0067]    In a further embodiment, the present invention provides polynucleotides that hybridise under stringent conditions to either  
         [0068]    (a) a DNA sequence encoding a polypeptide or  
         [0069]    (b) the complement of a DNA sequence encoding a polypeptide;  
         [0070]    wherein said polypeptide comprising a sequence chosen from SEQ ID NOs: 2, 4, 6, 8, 10, 12, 14, 16, 20 or fragments or analogues thereof.  
         [0071]    In a further embodiment, the present invention provides polynucleotides that hybridise under stringent conditions to either  
         [0072]    (a) a DNA sequence encoding a polypeptide or  
         [0073]    (b) the complement of a DNA sequence encoding a polypeptide;  
         [0074]    wherein said polypeptide comprises at least 10 contiguous amino acid residues from a polypeptide comprising a sequence chosen from SEQ ID NOs: 2, 4, 6, 8, 10, 12, 14, 16, 20 or fragments or analogues thereof.  
         [0075]    In a further embodiment, polynucleotides are those encoding polypeptides of the invention illustrated in SEQ ID NOs: 2, 4, 6, 8, 10, 12, 14, 16, 20.  
         [0076]    In a further embodiment, polynucleotides are those illustrated in SEQ ID NOs: 1, 3, 5, 7, 9, 11, 13, 15, 19 encoding polypeptides of the invention.  
         [0077]    As will be readily appreciated by one skilled in the art, polynucleotides include both DNA and RNA.  
         [0078]    The present invention also includes polynucleotides complementary to the polynucleotides described in the present application.  
         [0079]    In a further aspect, polynucleotides encoding polypeptides of the invention, or fragments, analogues or derivatives thereof, may be used in a DNA immunization method. That is, they can be incorporated into a vector which is replicable and expressible upon injection thereby producing the antigenic polypeptide in vivo. For example polynucleotides may be incorporated into a plasmid vector under the control of the CMV promoter which is functional in eukaryotic cells. Preferably the vector is injected intramuscularly.  
         [0080]    According to another aspect, there is provided a process for producing polypeptides of the invention by recombinant techniques by expressing a polynucleotide encoding said polypeptide in a host cell and recovering the expressed polypeptide product. Alternatively, the polypeptides can be produced according to established synthetic chemical techniques i.e. solution phase or solid phase synthesis of oligopeptides which are ligated to produce the full polypeptide (block ligation).  
         [0081]    General methods for obtention and evaluation of polynucleotides and polypeptides are described in the following references: Sambrook et al, Molecular Cloning: A Laboratory Manual, 2nd ed, Cold Spring Harbor, N.Y., 1989; Current Protocols in Molecular Biology, Edited by Ausubel F. M. et al., John Wiley and Sons, Inc. New York; PCR Cloning Protocols, from Molecular Cloning to Genetic Engineering, Edited by White B. A., Humana Press, Totowa, N.J., 1997, 490 pages; Protein Purification, Principles and Practices, Scopes R. K., Springer-Verlag, New York, 3rd Edition, 1993, 380 pages; Current Protocols in Immunology, Edited by Coligan J. E. et al., John Wiley &amp; Sons Inc., New York which are herein incorporated by reference.  
         [0082]    For recombinant production, host cells are transfected with vectors which encode the polypeptide, and then cultured in a nutrient media modified as appropriate for activating promoters, selecting transformants or amplifying the genes. Suitable vectors are those that are viable and replicable in the chosen host and include chromosomal, non-chromosomal and synthetic DNA sequences e.g. bacterial plasmids, phage DNA, baculovirus, yeast plasmids, vectors derived from combinations of plasmids and phage DNA. The polypeptide sequence may be incorporated in the vector at the appropriate site using restriction enzymes such that it is operably linked to an expression control region comprising a promoter, ribosome binding site (consensus region or Shine-Dalgarno sequence), and optionally an operator (control element). One can select individual components of the expression control region that are appropriate for a given host and vector according to established molecular biology principles (Sambrook et al, Molecular Cloning: A Laboratory Manual, 2nd ed, Cold Spring Harbor, N.Y., 1989; Current Protocols in Molecular Biology, Edited by Ausubel F. M. et al., John. Wiley and Sons, Inc. New York incorporated herein by reference). Suitable promoters include but are not limited to LTR or SV40 promoter,  E. coli  lac, tac or trp promoters and the phage lambda P L  promoter. Vectors will preferably incorporate an origin of replication as well as selection markers i.e. ampicilin resistance gene. Suitable bacterial vectors include pET, pQE70, pQE60, pQE-9, pbs, pD10 phagescript, psiX174, pbluescript SK, pbsks, pNH8A, pNH16a, pNH18A, pNH46A, ptrc99a, pKK223-3, pKK233-3, pDR540, pRIT5 and eukaryotic vectors pBlueBacIII, pWLNEO, pSV2CAT, pOG44, pXT1, pSG, pSVK3, pBPV, pMSG and pSVL. Host cells may be bacterial i.e.  E. coli, Bacillus subtilis,  Streptomyces; fungal i.e.  Aspergillus niger, Aspergillus nidulins;  yeast i.e. Saccharomyces or eukaryotic i.e. CHO, COS.  
         [0083]    Upon expression of the polypeptide in culture, cells are typically harvested by centrifugation then disrupted by physical or chemical means (if the expressed polypeptide is not secreted into the media) and the resulting crude extract retained to isolate the polypeptide of interest. Purification of the polypeptide from culture media or lysate may be achieved by established techniques depending on the properties of the polypeptide i.e. using ammonium sulfate or ethanol precipitation, acid extraction, anion or cation exchange chromatography, phosphocellulose chromatography, hydrophobic interaction chromatography, hydroxylapatite chromatography and lectin chromatography. Final purification may be achieved using HPLC.  
         [0084]    The polypeptide may be expressed with or without a leader or secretion sequence. In the former case the leader may be removed using post-translational processing (see U.S. Pat. No. 4,431,739; U.S. Pat. No. 4,425,437; and U.S. Pat. No. 4,338,397 incorporated herein by reference) or be chemically removed subsequent to purifying the expressed polypeptide.  
         [0085]    According to a further aspect, the streptococcal polypeptides of the invention may be used in a diagnostic test for streptococcus infection, in particular  Streptococcus pyogenes  infection. Several diagnostic methods are possible, for example detecting streptococcus organism in a biological sample, the following procedure may be followed:  
         [0086]    a) obtaining a biological sample from an individual;  
         [0087]    b) incubating an antibody or fragment thereof reactive with a streptococcus polypeptide of the invention with the biological sample to form a mixture; and  
         [0088]    c) detecting specifically bound antibody or bound fragment in the mixture which indicates the presence of streptococcus.  
         [0089]    Alternatively, a method for the detection of antibody specific to a streptococcus antigen in a biological sample containing or suspected of containing said antibody may be performed as follows:  
         [0090]    a) obtaining a biological sample from an individual;  
         [0091]    b) incubating one or more streptococcus polypeptides of the invention or fragments thereof with the biological sample to form a mixture; and  
         [0092]    c) detecting specifically bound antigen or bound fragment in the mixture which indicates the presence of antibody specific to streptococcus.  
         [0093]    One of skill in the art will recognize that this diagnostic test may take several forms, including an immunological test such as an enzyme-linked immunosorbent assay (ELISA), a radioimmunoassay or a latex agglutination assay, essentially to determine whether antibodies specific for the protein are present in an individual.  
         [0094]    The DNA sequences encoding polypeptides of the invention may also be used to design DNA probes for use in detecting the presence of streptococcus in a biological sample suspected of containing such bacteria. The detection method of this invention comprises:  
         [0095]    a) obtaining the biological sample from an individual;  
         [0096]    b) incubating one or more DNA probes having a DNA sequence encoding a polypeptide of the invention or fragments thereof with the biological sample to form a mixture; and  
         [0097]    c) detecting specifically bound DNA probe in the mixture which indicates the presence of streptococcus bacteria.  
         [0098]    The DNA probes of this invention may also be used for detecting circulating streptococcus i.e.  Streptococcus pyogenes  nucleic acids in a sample, for example using a polymerase chain reaction, as a method of diagnosing streptococcus infections. The probe may be synthesized using conventional techniques and may be immobilized on a solid phase, or may be labelled with a detectable label. A preferred DNA probe for this application is an oligomer having a sequence complementary to at least about 6 contiguous nucleotides of the  Streptococcus pyogenes  polypeptides of the invention.  
         [0099]    Another diagnostic method for the detection of streptococcus in an individual comprises:  
         [0100]    a) labelling an antibody reactive with a polypeptide of the invention or fragment thereof with a detectable label;  
         [0101]    b) administering the labelled antibody or labelled fragment to the patient; and  
         [0102]    c) detecting specifically bound labelled antibody or labelled fragment in the patient which indicates the presence of streptococcus.  
         [0103]    A further aspect of the invention is the use of the streptococcus polypeptides of the invention as immunogens for the production of specific antibodies for the diagnosis and in particular the treatment of streptococcus infection. Suitable antibodies may be determined using appropriate screening methods, for example by measuring the ability of a particular antibody to passively protect against streptococcus infection in a test model. One example of an animal model is the mouse model described in the examples herein. The antibody may be a whole antibody or an antigen-binding fragment thereof and may belong to any immunoglobulin class. The antibody or fragment may be of animal origin, specifically of mammalian origin and more specifically of murine, rat or human origin. It may be a natural antibody or a fragment thereof, or if desired, a recombinant antibody or antibody fragment. The term recombinant antibody or antibody fragment means antibody or antibody fragment which was produced using molecular biology techniques. The antibody or antibody fragments may be polyclonal, or preferably monoclonal. It may be specific for a number of epitopes associated with the  Streptococcus pyogenes  polypeptides but is preferably specific for one.  
         [0104]    A further aspect of the invention is the use of the antibodies directed to the streptococcus polypeptides of the invention for passive immunization. One could use the antibodies described in the present application. Suitable antibodies may be determined using appropriate screening methods, for example by measuring the ability of a particular antibody to passively protect against streptococcus infection in a test model. One example of an animal model is the mouse model described in the examples herein. The antibody may be a whole antibody or an antigen-binding fragment thereof and may belong to any immunoglobulin class. The antibody or fragment may be of animal origin, specifically of mammalian origin and more specifically of murine, rat or human origin. It may be a natural antibody or a fragment thereof, or if desired, a recombinant antibody or antibody fragment. The term recombinant antibody or antibody fragment means antibody or antibody fragment which was produced using molecular biology techniques. The antibody or antibody fragments may be polyclonal, or preferably monoclonal. It may be specific for a number of epitopes associated with the  streptococcus pneumoniae  polypeptides but is preferably specific for one.  
         [0105]    Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. All publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety. In case of conflict, the present specification, including definitions, will control. In addition, the materials, methods, and examples are illustrative only and not intended to be limiting.  
       EXAMPLE 1  
       [0106]    This example illustrates the cloning of  S. pyogenes  gene.  
         [0107]    The coding region of  S. pyogenes  gene BVH-P1 (SEQ ID NO:1) was amplified by PCR (DNA Thermal Cycler GeneAmp PCR system 2400 Perkin Elmer, San Jose, Calif.) from genomic DNA of serotype 3  S. pyogenes  strain ATCC12384 using the following oligos that contained base extensions for the addition of restriction sites NcoI (CCATGG) and XhoI (CTCGAG): DMAR16 (5′-CAGGCCATGGAGTGGACACCACGATCGGTTAC-3′); DMAR17 (5′-GCCGCTCGAGAGCATTAAAGGAGACATGAACATGATC-3′). PCR products were purified from agarose gel using a QIAquick gel extraction kit from QIAgen following the manufacturer&#39;s instructions (Chatsworth, Calif.), and digested with NcoI and XhoI (Pharmacia Canada Inc, Baie d&#39;Urfé, Canada). The pET-21d(+) vector (Novagen, Madison, Wis.) was digested with NcoI and XhoI and purified from agarose gel using a QIAquick gel extraction kit from QIAgen (Chatsworth, Calif.). The NcoI-XhoI PCR products were ligated to the NcoI-XhoI pET-21d(+)expression vector. The ligated products were transformed into  E. coli  strain  E. coli  strain DH5α [φ80dlacZΔM15 Δ(lacZYA-argF) U169 endA1 recA1 hsdR17(r K -m K +) deoR thi-1 supE44 λ − gyrA96 relA1] (Gibco BRL, Gaithersburg, Md.) according to the method of Simanis (Hanahan, D. DNA Cloning, 1985, D. M. Glover (ed), pp. 109-135). Recombinant pET-21d(+)plasmid (rpET21d(+)) containing BVH-P1 gene was purified using a QIAgen plasmid kit (Chatsworth, Calif.) and DNA insert was sequenced (Taq Dye Deoxy Terminator Cycle Sequencing kit, ABI, Foster City, Calif.).  
         [0108]    It was determined that the open reading frame (ORF) which codes for BVH-P1 contains 1170-bp and encodes a 389 amino acid residues polypeptide with a predicted pI of 4.37 and a predicted molecular mass of 41054 Da.  
         [0109]    Analysis of the predicted amino acid residues sequence (SEQ ID NO:2)using the Spscan sofware (Wisconsin Sequence Analysis Package; Genetics Computer Group) suggested the existence of a 25 amino acid residues signal peptide (MIITKKSLFVTSVALSLAPLATAQA), which ends with a cleavage site situated between an alanine and a glutamine residues. Analysis of this ORF did not revealed the presence of repetitive structures, cell wall anchoring motif (LPXTG), or IgA binding motif (MLKKIE).  
         [0110]    An ORF which shares 62% with the  S. pyogenes  BVH-P1 gene was initially presented in the patent application PCT/CA99/00114 which described Group B streptococcus antigens. BVH-PL gene was also found to share homology (62% identity) with an ORF present in the genome of  S. pneumoniae  (The Institute for Genomic Research).  
       EXAMPLE 2  
       [0111]    This example describes the PCR amplification and sequencing of BVH-P1 gene from other  S. pyogenes  strains and the evaluation of the level of molecular conservation of this gene.  
         [0112]    Lancefield&#39;s serogroup A  S. pyogenes  LSPQ2296 (ATCC 19615) was provided by the laboratoire de la santé publique du Québec, Sainte-Anne-de-Bellevue; serotype 1  S. pyogenes  SPY57 clinical isolate was provided by the centre de recherche en infectiologie du centre hospitalier de l&#39;universit{acute over (e )} Laval, Sainte-Foy; and  S. pyogenes  strain B514 which was initially isolated from a mouse was provided by Susan Hollingshead, from University of Alabama, Birmingham. The respective coding region of  S. pyogenes  gene BVH-P1 from strains ATCC 12384 (SEQ ID NO:1), LSPQ2699(ATCC19615) (SEQ ID NO:3), SPY57 (SEQ ID NO:5), and B514 (SEQ ID NO:7) were amplified by PCR (DNA Thermal Cycler GeneAmp PCR system 2400 Perkin Elmer, San Jose, Calif.) from bacterial cell lysates using the following oligos DMAR69 (5′-CTGGGAAGATTATCTAGCACATTAATAC-3′); DMAR72 (5′-CATAACGTTAAAACTGTCTAAAGGG-3′). PCR products were purified from agarose gel using a QIAquick gel extraction kit from QIAgen following the manufacturer&#39;s instructions (Chatsworth, Calif.) and the DNA insert were sequenced (Taq Dye Deoxy Terminator Cycle Sequencing kit, ABI, Foster City, Calif.). The predicted amino acid sequences from strains ATCC12384 (SEQ ID NO:2), LSPQ2699(ATCC19615) (SEQ ID NO:4), SPY57 (SEQ ID NO:6), and B514 (SEQ ID NO:8) were respectively presented in the following FIGS. 2, 4,  6 , and  8 .  
         [0113]    The FIGS. 17 and 18 respectively depict the consensus nucleotide and predicted amino acid sequences established for  S. pyogenes  BVH-P1. In addition to the sequences presented herewith, the BVH-P1 gene sequences from the genome sequencing project at the University of Oklahoma (serotype M1  S. pyogenes  strain ATCC 70029: http://dnal.chem.ou.edu/strep.html) and from (Kil et al. 1994. Infect. Immun. 62:2440-2449: GenBank accession number U09352) were also included. No function or role in the pathogenesis of the bacteria or protection against infection was described by Kil et al. for the sequence with GenBank accession number U09352. This latter sequence presented by Kil et al. was shown to be located upstream of a  S. pyogenes  67 kDa myosin-cross-reactive antigen.  
         [0114]    Pairwise comparison of the BVH-P1 predicted protein sequences revealed between 95 to 100% identity with the exception of the BVH-P1 sequence obtained from GenBank under the accesssion number U09352. Pairwise comparison of that particular sequence with the other five BVH-P1 sequences indicated identity between 87 to 91%. This lower homology can be explained by the presence of two regions (119-124 and 262-281) which are more divergent comparatively to the other BVH-P1 gene sequences. Beside these two regions in the BVH-P1 sequence obtained from GenBank under the accesssion number U09352, the BVH-P1 genes showed great similarity in overall organization.  
       EXAMPLE 3  
       [0115]    This example illustrates the cloning of  S. pyogenes  protein gene in CMV plasmid pCMV-GH.  
         [0116]    The DNA coding region of a  S. pyogenes  protein was inserted in phase downstream of a human growth hormone (hGH) gene which was under the transcriptional control of the cytomegalovirus (CMV) promotor in the plasmid vector pCMV-GH (Tang et al., Nature, 1992, 356:152). The CMV promotor is a non functional plasmid in  E. coli  cells but is active upon administration of the plasmid in eukaryotic cells. The vector also incorporated the ampicillin resistance gene.  
         [0117]    The coding region of BVH-P1 gene (SEQ ID NO:9) without its leader peptide region was amplified by PCR (DNA Thermal Cycler GeneAmp PCR system 2400 Perkin Elmer, San Jose, Calif.) from genomic DNA of serotype 3  S. pyogenes  strain ATCC12384 using the following oligos that contained base extensions for the addition of restriction sites BamHI (GGATCC) and SalI (GTCGAC): DMAR24 (5′-TACCCGGATCCCCAAGAGTGGACACCACGATCGG-3′); DMAR25 (5′-GCGCTCGTCGACGCGTATCTCAGCCTCTTATAGGGC-3′). The PCR product was purified from agarose gel using a QIAquick gel extraction kit from QIAgen (Chatsworth, Calif.), digested with restriction enzymes (Pharmacia Canada Inc, Baie d&#39;Urfe, Canada). The pCMV-GH vector (Laboratory of Dr. Stephen A. Johnston, Department of Biochemistry, The University of Texas, Dallas, Tex.) was digested with BamHI and SalI and purified from agarose gel using the QIAquick gel extraction kit from QIAgen (Chatsworth, Calif.). The BamHI-SalI DNA fragments were ligated to the BamHI-SalI pCMV-GH vector to create the hGH-BVH-P1 fusion protein under the control of the CMV promoter. The ligated products were transformed into  E. coli  strain DH5α [φ80dlacZΔM15 Δ(lacZYA-argF)U169 endA1 recA1 hsdR17(r K -m K +) deoR thi-1 supE44 λ − gyrA96 relA1] (Gibco BRL, Gaithersburg, Md.) according to the method of Simanis (Hanahan, D. DNA Cloning, 1985, D. M. Glover (ed), pp. 109-135). The recombinant pCMV plasmid was purified using a QIAgen plasmid kit (Chatsworth, Calif.) and the nucleotide sequence of the DNA insert was verified by DNA sequencing.  
       EXAMPLE 4  
       [0118]    This example illustrates the use of DNA to elicit an immune response to  S. pyogenes  antigens.  
         [0119]    A group of 8 female BALB/c mice (Charles River, St-Constant, Québec, Canada) were immunized by intramuscular injection of 100 μl three times at two- or three-week intervals with 50 μg of recombinant PCMV-GH encoding BVH-P1 gene in presence of 50 μg of granulocyte-macrophage colony-stimulating factor (GM-CSF)-expressing plasmid pCMV-GH-GM-CSF (Laboratory of Dr. Stephen A. Johnston, Department of Biochemistry, The University of Texas, Dallas, Tex.). As control, a group of mice were injected with 50 μg of pCMV-GH in presence of 50 μg of pCMV-GH-GM-CSF. Blood samples were collected from the orbital sinus prior to each immunization and seven days following the third injection and serum antibody responses were determined by ELISA using purified BVH-P1-His•Tag from SEQ ID NO:11  S. pyogenes  recombinant protein as coating antigen.  
       EXAMPLE 5  
       [0120]    This example illustrates the production and purification of recombinant  S. pyogenes  BVH-P1 protein.  
         [0121]    The recombinant pET-21d(+)plasmid with BVH-P1 gene corresponding to the SEQ ID NO:9 was used to transform by electroporation (Gene Pulser II apparatus, BIO-RAD Labs, Mississauga, Canada)  E. coli  strain BL21(DE3) (F − ompT hsdS B  (r −   B m −   B ) gal dcm (DE3)) (Novagen, Madison, Wis.). In this strain of  E. coli,  the T7 promotor controlling expression of the recombinant protein is specifically recognized by the T7 RNA polymerase (present on the λDE3 prophage) whose gene is under the control of the lac promotor which is inducible by isopropyl-β-d-thio-galactopyranoside (IPTG). The transformant BL21(DE3)/rpET was grown at 37° C. with agitation at 250 rpm in LB broth (peptone 10 g/L, yeast extract 5 g/L, NaCl 10 g/L) containing 100 μg of carbenicillin (Sigma-Aldrich Canada Ltd., Oakville, Canada) per ml until the A 600  reached a value of 0.6. In order to induce the production of  S. pyogenes  BVH-P1-His•Tag recombinant protein (from SEQ ID NO:10), the cells were incubated for 3 additional hours in the presence of IPTG at a final concentration of 1 mM. Induced cells from a 500 ml culture were pelleted by centrifugation and frozen at −70° C.  
         [0122]    The purification of the recombinant proteins from the soluble cytoplasmic fraction of IPTG-induced BL21(DE3)/rpET21b(+) was done by affinity chromatography based on the properties of the His•Tag sequence (6 consecutive histidine residues) to bind to divalent cations (Ni 2+ ) immobilized on the His•Bind metal chelation resin. Briefly, the pelleted cells obtained from a 500 mL culture induced with IPTG was resuspended in lysis buffer (20 mM Tris, 500 mM NaCl. 10 mM imidazole, pH 7.9) containing 1 mM PMSF, sonicated and centrifuged at 12,000×g for 20 min to remove debris. The supernatant was deposited on a Ni-NTA agarose column (Qiagen, Mississauga, Ontario, Canada). The  S. pyogenes  BVH-P1-His•Tag recombinant protein (from SEQ ID NO:10) was eluted with 250 mM imidazole-500 mM NaCl-20 mM Tris pH 7.9. The removal of the salt and imidazole from the sample was done by dialysis against PBS at 4° C. The quantities of recombinant protein obtained from the soluble fraction of  E. coli  was estimated by MicroBCA (Pierce, Rockford, Ill.).  
       EXAMPLE 6  
       [0123]    This example illustrates the accessibility to antibodies of the BVH-P1 protein at the surface of  S. pyogenes  strain.  
         [0124]    Bacteria were grown in Tood Hewitt (TH) broth (Difco Laboratories, Detroit Mich.) with 0.5% Yeast extract (Difco Laboratories) and 0.5% peptone extract (Merck, Darmstadt, Germany) at 37° C. in a 8% CO 2  atmosphere to give an OD 490nm  of 0.600 (˜10 8  CFU/ml). Dilutions of anti-BVH-P1 or control sera were then added and allowed to bind to the cells, which were incubated for 2 h at 4° C. Samples were washed 4 times in blocking buffer [phosphate-buffered saline (PBS) containing 2% bovine serum albumin (BSA)], and then 1 ml of goat fluorescein (FITC)-conjugated anti-mouse IgG+IgM diluted in blocking buffer was added. After an additional incubation of 60 min at room temperature, samples were washed 4 times in blocking buffer and fixed with 0.25% formaldehyde in PBS buffer for 18-24 h at 4° C. Cells were washed 2 times in PBS buffer and resuspended in 500 μl of PBS buffer. Cells were kept in the dark at 4° C. until analyzed by flow cytometry (Epics® XL; Beckman Coulter, Inc.). Flow cytometric analysis revealed that BVH-P1-specific antibodies efficiently recognized their corresponding surface exposed epitopes on both the homologous (ATCC12384; serotype3) and the heterologous (SPY57; seotype 1)  S. pyogenes  strains tested. It was determined that more than 90% of the 10,000  S. pyogenes  cells analyzed were labeled with the antobodies present in the BVH-MC1 specific anti-sera. These observations clearly demonstrate that the BVH-P1 protein is accessible at the surface where it can be easily recognized by antibodies. Anti- S. pyogenes  antibodies were shown to play an important role in the protection against  S. pyogenes  infection.  
       EXAMPLE 7  
       [0125]    This example illustrates the protection against fatal  S. pyogenes  infection induced by passive immunization of mice with rabbit hyper-immune sera.  
         [0126]    New Zealand rabbits (Charles River laboratories, Montreal, Canada) were injected subcutaneously at multiple sites with approximately 50 μg and 100 μg of BVH-P1-His•Tag protein (from SEQ ID NO:10) that was produced and purified as described in Example 5 and adsorbed to Alhydrogel adjuvant (Superfos Biosector a/s). Rabbits were immunized three times at three-week intervals with the BVH-P1-His•Tag protein (from SEQ ID NO:10). Blood samples were collected three weeks after the third injection. The antibodies present in the serum were purified by precipitation using 40% saturated ammonium sulfate. Groups of 10 female CD-1 mice (Charles River) were injected intravenously with 500 μl of purified serum collected either from BVH-P1-His•Tag (from SEQ ID NO:10) immunized rabbits or rabbits immunized with an unrelated control recombinant protein. Eighteen hours later the mice were challenged with approximately 2×10 7  CFU of the type 3  S. pyogenes  strain ATCC12384. Samples of the  S. pyogenes  challenge inoculum were plated on blood agar plates to determine the CFU and to verify the challenge dose. Deaths were recorded for a period of 5 days.  
       EXAMPLE 8  
       [0127]    This example illustrates the protection of mice against fatal  S. pyogenes  infection induced by immunization with BVH-P1 protein.  
         [0128]    Groups of 8 female CD-1 mice (Charles River) were immunized subcutaneously three times at three-week intervals with 20 μg of affinity purified  S. pyogenes  BVH-P1-His•Tag recombinant protein (from SEQ ID NO:10) in presence of 10 μg of QuilA adjuvant (Cedarlane Laboratories Ltd, Hornby, Canada) or, as control, with QuilA adjuvant alone in PBS. Blood samples were collected from the orbital sinus on day 1, 22 and 43 prior to each immunization and seven days (day 50) following the third injection. Analysis by ELISA using purified recombinant BVH-P1 protein (from SEQ ID NO:10) clearly indicated that this protein is highly immunogenic in animals. Indeed reciprocal ELISA titers higher than 10 6  were determined for the mice immunized with this recombinant protein. Two weeks later the mice were challenged with approximately 2×10 7  CFU of the type 3  S. pyogenes  strain ATCC12384. Samples of the  S. pyogenes  challenge inoculum were plated on blood agar plates to determine the CFU and to verify the challenge dose. Deaths were recorded for a period of 5 days. Five out of the 8 (62%) mice immunized with three injections of 20 μg of purified recombinant BVH-P1 (from SEQ ID NO:10) and QuilA adjuvant survived the bacterial challenge to only 2/7 (28%)in the control group.  
                                                           TABLE 3                           Immunization of CD-1 mice with purified recombinant       BVH-P1 protein confers protection against subsequent challenge       with  S. pyogenes  strain ATCC 12384                    Survival of the mice challenged with  S.                   pyogenes  strain ATCC 12384 (Day after               challenge: number of survivors/total               number of mice challenged))                Groups   1   2   3   4   5                       20 μg of   8/8   8/8   7/8   6/8   5/8           BVH-P1-           His•Tag           Control   7/7   6/7   3/7   2/7   2/7                      
 
         [0129]    [0129] 
     
       
       
         1 
         
           
             29  
           
           
             1  
             1170  
             DNA  
             S. pyogenes  
           
            1 

atgattatta ctaaaaagag cttatttgtg acaagtgtcg ctttgtcgtt agcacctttg     60 

gcgacagcac aggcacaaga gtggacacca cgatcggtta cagaaatcaa gtctgaactc    120 

gtcctagttg ataatgtttt tacttatact gtaaaatacg gtgacacttt aagcacaatt    180 

gctgaagcaa tgggaattga tgtgcatgtc ttaggagata ttaatcatat tgctaatatt    240 

gacttaattt ttccagacac gatcctaaca gccaactaca accaacacgg tcaggcaacg    300 

actttgacgg ttcaagcgcc tgcttctagt ccagctagcg ttagtcatgt acctagcagt    360 

gagccattac cccaagcatc tgccacctct caatcgactg ttcctatggc accatctgcg    420 

acaccatctg atgtcccaac gacaccattc gcatctgcaa agccagatag ttctgtgaca    480 

gcgtcatctg agctcacatc gtcaacgaat gatgtttcga ctgagttgtc tagcgaatca    540 

caaaagcagc cagaagtacc acaagaagca gttccaactc ctaaagcagc tgaaacgact    600 

gaagtcgaac ctaagacaga catctcagag gattcaactt cagctaatag gcctgtacct    660 

aacgagagtg cttcagaaga agtttcttct gcggccccag cacaagcccc agcagaaaaa    720 

gaagaaacct ctgcgccagc agcacaaaaa gctgtagctg acaccacaag tgttgcaacc    780 

tcaaatggcc tttcttacgc tccaaaccat gcctacaatc caatgaatgc agggcttcaa    840 

ccacaaacag cagccttcaa agaagaagtg gcttctgcct ttggtattac gtcatttagt    900 

ggttaccgtc caggtgatcc aggagatcat ggtaaaggtt tggccattga ttttatggtg    960 

cctgaaaatt ctgctcttgg tgatcaagtt gctcaatatg ccattgacca tatggcagag   1020 

cgtggtattt catacgttat ttggaaacag cgattctatg cgccatttgc aagtatttac   1080 

ggaccagcct acacatggaa ccccatgcca gatcgcggca gtattacaga aaaccattat   1140 

gatcatgttc atgtctcctt taatgcttaa                                    1170 

 
           
             2  
             389  
             PRT  
             S. pyogenes  
           
            2 

Met Ile Ile Thr Lys Lys Ser Leu Phe Val Thr Ser Val Ala Leu Ser 
 1               5                  10                  15 

Leu Ala Pro Leu Ala Thr Ala Gln Ala Gln Glu Trp Thr Pro Arg Ser 
            20                  25                  30 

Val Thr Glu Ile Lys Ser Glu Leu Val Leu Val Asp Asn Val Phe Thr 
        35                  40                  45 

Tyr Thr Val Lys Tyr Gly Asp Thr Leu Ser Thr Ile Ala Glu Ala Met 
    50                  55                  60 

Gly Ile Asp Val His Val Leu Gly Asp Ile Asn His Ile Ala Asn Ile 
65                  70                  75                  80 

Asp Leu Ile Phe Pro Asp Thr Ile Leu Thr Ala Asn Tyr Asn Gln His 
                85                  90                  95 

Gly Gln Ala Thr Thr Leu Thr Val Gln Ala Pro Ala Ser Ser Pro Ala 
            100                 105                 110 

Ser Val Ser His Val Pro Ser Ser Glu Pro Leu Pro Gln Ala Ser Ala 
        115                 120                 125 

Thr Ser Gln Ser Thr Val Pro Met Ala Pro Ser Ala Thr Pro Ser Asp 
    130                 135                 140 

Val Pro Thr Thr Pro Phe Ala Ser Ala Lys Pro Asp Ser Ser Val Thr 
145                 150                 155                 160 

Ala Ser Ser Glu Leu Thr Ser Ser Thr Asn Asp Val Ser Thr Glu Leu 
                165                 170                 175 

Ser Ser Glu Ser Gln Lys Gln Pro Glu Val Pro Gln Glu Ala Val Pro 
            180                 185                 190 

Thr Pro Lys Ala Ala Glu Thr Thr Glu Val Glu Pro Lys Thr Asp Ile 
        195                 200                 205 

Ser Glu Asp Ser Thr Ser Ala Asn Arg Pro Val Pro Asn Glu Ser Ala 
    210                 215                 220 

Ser Glu Glu Val Ser Ser Ala Ala Pro Ala Gln Ala Pro Ala Glu Lys 
225                 230                 235                 240 

Glu Glu Thr Ser Ala Pro Ala Ala Gln Lys Ala Val Ala Asp Thr Thr 
                245                 250                 255 

Ser Val Ala Thr Ser Asn Gly Leu Ser Tyr Ala Pro Asn His Ala Tyr 
            260                 265                 270 

Asn Pro Met Asn Ala Gly Leu Gln Pro Gln Thr Ala Ala Phe Lys Glu 
        275                 280                 285 

Glu Val Ala Ser Ala Phe Gly Ile Thr Ser Phe Ser Gly Tyr Arg Pro 
    290                 295                 300 

Gly Asp Pro Gly Asp His Gly Lys Gly Leu Ala Ile Asp Phe Met Val 
305                 310                 315                 320 

Pro Glu Asn Ser Ala Leu Gly Asp Gln Val Ala Gln Tyr Ala Ile Asp 
                325                 330                 335 

His Met Ala Glu Arg Gly Ile Ser Tyr Val Ile Trp Lys Gln Arg Phe 
            340                 345                 350 

Tyr Ala Pro Phe Ala Ser Ile Tyr Gly Pro Ala Tyr Thr Trp Asn Pro 
        355                 360                 365 

Met Pro Asp Arg Gly Ser Ile Thr Glu Asn His Tyr Asp His Val His 
    370                 375                 380 

Val Ser Phe Asn Ala 
385 

 
           
             3  
             1182  
             DNA  
             S. pyogenes  
           
            3 

atgattatta ctaaaaagag cttatttgtg acaagtgtcg ctttgtcgtt agcacctttg     60 

gcgacagcgc aggcacaaga gtggacacca cgatcggtta cagaaatcaa gtctgaactc    120 

gtcctagttg ataatgtttt tacttatata gtaaaatacg gtgacacttt aagcacaatt    180 

gctgaagcaa tggggattga tgtgcatgtc ttaggagata ttaatcatat tgctaatatt    240 

gacttaattt ttccagacac gatcctaaca gcaaactaca accaacacgg tcaggcaacg    300 

actttgacgg ttcaagcacc tgcttctagt ccatctagcg ttagtcatgt acctagcagt    360 

gagccattac cccaagcatc tgccacctct caaccgactg ttcctatggc accatctgcg    420 

acaccatctg atgtcccaac gacaccattc gcatctgcaa agccagatag ttctgtgaca    480 

gcgtcatctg agctcacatc gtcaacgaat gatgtttcga ctgagttgtc tagcgaatca    540 

caaaagcagc cagaagtacc acaagaagca gttccaactc ctaaagcagc tgaaccgact    600 

gaagtcgaac ctaagacaga catctcagaa gacccaactt cagctaatag gcctgtacct    660 

aacgagagtg cttcagaaga agcttcttct gcggccccag cacaagctcc agcagaaaaa    720 

gaagaaacct ctcagatgtt aactgcgcca gcagcacaaa aagctgtagc tgacaccaca    780 

agtgttgcaa cctcaaacgg cctttcttac gctccaaacc atgcctacaa tccaatgaat    840 

gcagggcttc aaccacaaac agcagccttc aaagaagaag tggcttctgc ctttggtatt    900 

acgtcattta gtggttaccg tccaggagat ccaggagatc atggtaaagg attagccatt    960 

gactttatgg taccggttag ctctacgctt ggtgatcaag ttgctcaata tgccattgac    020 

catatggcag agcgtggtat ttcatacgtt atttggaaac agcgattcta tgcgccattt    080 

gcaagtattt acggaccagc ctacacatgg aaccccatgc cagatcgcgg cagtattaca    140 

gaaaaccatt atgatcatgt tcatgtctcc tttaatgctt aa                       182 

 
           
             4  
             393  
             PRT  
             S. pyogenes  
           
            4 

Met Ile Ile Thr Lys Lys Ser Leu Phe Val Thr Ser Val Ala Leu Ser 
 1               5                  10                  15 

Leu Ala Pro Leu Ala Thr Ala Gln Ala Gln Glu Trp Thr Pro Arg Ser 
            20                  25                  30 

Val Thr Glu Ile Lys Ser Glu Leu Val Leu Val Asp Asn Val Phe Thr 
        35                  40                  45 

Tyr Ile Val Lys Tyr Gly Asp Thr Leu Ser Thr Ile Ala Glu Ala Met 
    50                  55                  60 

Gly Ile Asp Val His Val Leu Gly Asp Ile Asn His Ile Ala Asn Ile 
65                  70                  75                  80 

Asp Leu Ile Phe Pro Asp Thr Ile Leu Thr Ala Asn Tyr Asn Gln His 
                85                  90                  95 

Gly Gln Ala Thr Thr Leu Thr Val Gln Ala Pro Ala Ser Ser Pro Ser 
            100                 105                 110 

Ser Val Ser His Val Pro Ser Ser Glu Pro Leu Pro Gln Ala Ser Ala 
        115                 120                 125 

Thr Ser Gln Pro Thr Val Pro Met Ala Pro Ser Ala Thr Pro Ser Asp 
    130                 135                 140 

Val Pro Thr Thr Pro Phe Ala Ser Ala Lys Pro Asp Ser Ser Val Thr 
145                 150                 155                 160 

Ala Ser Ser Glu Leu Thr Ser Ser Thr Asn Asp Val Ser Thr Glu Leu 
                165                 170                 175 

Ser Ser Glu Ser Gln Lys Gln Pro Glu Val Pro Gln Glu Ala Val Pro 
            180                 185                 190 

Thr Pro Lys Ala Ala Glu Pro Thr Glu Val Glu Pro Lys Thr Asp Ile 
        195                 200                 205 

Ser Glu Asp Pro Thr Ser Ala Asn Arg Pro Val Pro Asn Glu Ser Ala 
    210                 215                 220 

Ser Glu Glu Ala Ser Ser Ala Ala Pro Ala Gln Ala Pro Ala Glu Lys 
225                 230                 235                 240 

Glu Glu Thr Ser Gln Met Leu Thr Ala Pro Ala Ala Gln Lys Ala Val 
                245                 250                 255 

Ala Asp Thr Thr Ser Val Ala Thr Ser Asn Gly Leu Ser Tyr Ala Pro 
            260                 265                 270 

Asn His Ala Tyr Asn Pro Met Asn Ala Gly Leu Gln Pro Gln Thr Ala 
        275                 280                 285 

Ala Phe Lys Glu Glu Val Ala Ser Ala Phe Gly Ile Thr Ser Phe Ser 
    290                 295                 300 

Gly Tyr Arg Pro Gly Asp Pro Gly Asp His Gly Lys Gly Leu Ala Ile 
305                 310                 315                 320 

Asp Phe Met Val Pro Val Ser Ser Thr Leu Gly Asp Gln Val Ala Gln 
                325                 330                 335 

Tyr Ala Ile Asp His Met Ala Glu Arg Gly Ile Ser Tyr Val Ile Trp 
            340                 345                 350 

Lys Gln Arg Phe Tyr Ala Pro Phe Ala Ser Ile Tyr Gly Pro Ala Tyr 
        355                 360                 365 

Thr Trp Asn Pro Met Pro Asp Arg Gly Ser Ile Thr Glu Asn His Tyr 
    370                 375                 380 

Asp His Val His Val Ser Phe Asn Ala 
385                 390 

 
           
             5  
             1170  
             DNA  
             S. pyogenes  
           
            5 

atgattatta ctaaaaagag cttatttgtg acaagtgtcg ctttgtcgtt agtacctttg     60 

gcgacagcgc aggcacaaga gtggacacca cgatcggtta cagaaatcaa gtctgaactc    120 

gtcctagttg ataatgtttt tacttatact gtaaaatacg gtgacacttt aagcacaatt    180 

gctgaagcaa tggggattga tgtgcatgtc ttaggagata ttaatcatat tgctaatatt    240 

gacctaattt ttccagacac gatcctaaca gcaaactaca atcaacacgg tcaggcaacg    300 

aatttgacgg ttcaagcacc tgcttctagt ccagctagcg ttagtcatgt acctagcagt    360 

gagccattac cccaagcatc tgccacctct caaccgactg ttcctatggc accacctgcg    420 

acaccatctg atgtcccaac gacaccattc gcatctgcaa agccagatag ttctgtgaca    480 

gcgtcatctg agctcacatc gtcaacgaat gatgtttcga ctgagttgtc tagcgaatca    540 

caaaagcagc cagaagtacc acaagaagca gttccaactc ctaaagcagc tgaaacgact    600 

gaagtcgaac ctaagacaga catctcagaa gccccaactt cagctaatag gcctgtacct    660 

aacgagagtg cttcagaaga agtttcttct gcggccccag cacaagcccc agcagaaaaa    720 

gaagaaacct ctgcgccagc agcacaaaaa gctgtagctg acaccacaag tgttgcaacc    780 

tcaaatggcc tttcttacgc tccaaaccat gcctacaatc caatgaatgc agggcttcaa    840 

ccacaaacag cagccttcaa agaagaagtg gcttctgcct ttggtattac gtcatttagt    900 

ggttaccgtc caggtgatcc aggagatcat ggtaaaggtt tggccattga ttttatggtg    960 

cgtggtattt catacgttat ttggaaacag cgattctatg cgccatttgc aagtatttac   1080 

ggaccagcct acacatggaa ccccatgcca gatcgcggca gtattacaga aaaccattat   1140 

gatcatgttc atgtctcctt taatgcttaa                                    1170 

 
           
             6  
             389  
             PRT  
             S. pyogenes  
           
            6 

Met Ile Ile Thr Lys Lys Ser Leu Phe Val Thr Ser Val Ala Leu Ser 
 1               5                  10                  15 

Leu Val Pro Leu Ala Thr Ala Gln Ala Gln Glu Trp Thr Pro Arg Ser 
            20                  25                  30 

Val Thr Glu Ile Lys Ser Glu Leu Val Leu Val Asp Asn Val Phe Thr 
        35                  40                  45 

Tyr Thr Val Lys Tyr Gly Asp Thr Leu Ser Thr Ile Ala Glu Ala Met 
    50                  55                  60 

Gly Ile Asp Val His Val Leu Gly Asp Ile Asn His Ile Ala Asn Ile 
65                  70                  75                  80 

Asp Leu Ile Phe Pro Asp Thr Ile Leu Thr Ala Asn Tyr Asn Gln His 
                85                  90                  95 

Gly Gln Ala Thr Asn Leu Thr Val Gln Ala Pro Ala Ser Ser Pro Ala 
            100                 105                 110 

Ser Val Ser His Val Pro Ser Ser Glu Pro Leu Pro Gln Ala Ser Ala 
        115                 120                 125 

Thr Ser Gln Pro Thr Val Pro Met Ala Pro Pro Ala Thr Pro Ser Asp 
    130                 135                 140 

Val Pro Thr Thr Pro Phe Ala Ser Ala Lys Pro Asp Ser Ser Val Thr 
145                 150                 155                 160 

Ala Ser Ser Glu Leu Thr Ser Ser Thr Asn Asp Val Ser Thr Glu Leu 
                165                 170                 175 

Ser Ser Glu Ser Gln Lys Gln Pro Glu Val Pro Gln Glu Ala Val Pro 
            180                 185                 190 

Thr Pro Lys Ala Ala Glu Thr Thr Glu Val Glu Pro Lys Thr Asp Ile 
        195                 200                 205 

Ser Glu Ala Pro Thr Ser Ala Asn Arg Pro Val Pro Asn Glu Ser Ala 
    210                 215                 220 

Ser Glu Glu Val Ser Ser Ala Ala Pro Ala Gln Ala Pro Ala Glu Lys 
225                 230                 235                 240 

Glu Glu Thr Ser Ala Pro Ala Ala Gln Lys Ala Val Ala Asp Thr Thr 
                245                 250                 255 

Ser Val Ala Thr Ser Asn Gly Leu Ser Tyr Ala Pro Asn His Ala Tyr 
            260                 265                 270 

Asn Pro Met Asn Ala Gly Leu Gln Pro Gln Thr Ala Ala Phe Lys Glu 
        275                 280                 285 

Glu Val Ala Ser Ala Phe Gly Ile Thr Ser Phe Ser Gly Tyr Arg Pro 
    290                 295                 300 

Gly Asp Pro Gly Asp His Gly Lys Gly Leu Ala Ile Asp Phe Met Val 
305                 310                 315                 320 

Pro Glu Asn Ser Ala Leu Gly Asp Gln Val Ala Gln Tyr Ala Ile Asp 
                325                 330                 335 

His Met Ala Glu Arg Gly Ile Ser Tyr Val Ile Trp Lys Gln Arg Phe 
            340                 345                 350 

Tyr Ala Pro Phe Ala Ser Ile Tyr Gly Pro Ala Tyr Thr Trp Asn Pro 
        355                 360                 365 

Met Pro Asp Arg Gly Ser Ile Thr Glu Asn His Tyr Asp His Val His 
    370                 375                 380 

Val Ser Phe Asn Ala 
385 

 
           
             7  
             1149  
             DNA  
             S. pyogenes  
           
            7 

atgattatta ctaaaaagag cttatttgtg acaagtgtcg ctttgtcgtt agcacctttg     60 

gcgacagcgc aggcacaaga gtggacacca cgatcggtta cagaaatcaa gtctgaactc    120 

gtcctagttg ataatgtttt tacttataca gtaaaatacg gtgacacttt aagcacaatt    180 

gctgaagcaa tggggattga tgtgcatgtc ttaggagata ttaatcatat tgctaatatt    240 

gacttaattt ttccagacac gatcctaaca gcaaactaca atcaacacgg tcaggcaacg    300 

actttgacgg ttcaagcacc tgcttctagt ccagctagcg ttagtcatgt acctagcagt    360 

gagccattac cccaagcatc tgccacctct caaccgactg ttcctatggc accatctgcg    420 

acaccattag catctgcaaa gccagatagt tctgtgacag cgtcatctga gctcacatcg    480 

tcaacgaatg atgtttcgac tgagtcgtct agcgaatcac aaaagcagcc agaagtacca    540 

caagaagcag ttccaactcc taaagcagct gaaacgactg aagtcgaacc taagacagac    600 

atctcagaag acccaacttc agctaatagg cctgtaccta acgagagtgc ttcagaagaa    660 

gtttcttctg cggccccagc acaagcccca gcagaaaaag aagaaacctc tgcgccagca    720 

gcacaaaaag ctgtagctga caccacaagt gttgcaacct caaacggcct ttcttacgct    780 

ccaaaccatg cctacaatcc aatgaatgca gggcttcaac cacaaacagc agccttcaaa    840 

gaagaagtgg cttctgcctt tggtattacg tcatttagtg gttaccgtcc aggtgaccca    900 

ggagatcatg gtaaaggttt ggccattgat tttatggtgc ctgaaaattc tgctcttggt    960 

gatcaagttg ctcaatatgc cattgaccat atggcagagc gtggtatttc atacgttatt   1020 

tggaaacagc gattctatgc gccatttgca agtatttacg gaccagctta cacatggaac   1080 

cccatgccag atcgcggcag tattacagaa aaccattatg atcatgttca tgtctccttt   1140 

aatgcttaa                                                           1149 

 
           
             8  
             382  
             PRT  
             S. pyogenes  
           
            8 

Met Ile Ile Thr Lys Lys Ser Leu Phe Val Thr Ser Val Ala Leu Ser 
 1               5                  10                  15 

Leu Ala Pro Leu Ala Thr Ala Gln Ala Gln Glu Trp Thr Pro Arg Ser 
            20                  25                  30 

Val Thr Glu Ile Lys Ser Glu Leu Val Leu Val Asp Asn Val Phe Thr 
        35                  40                  45 

Tyr Thr Val Lys Tyr Gly Asp Thr Leu Ser Thr Ile Ala Glu Ala Met 
    50                  55                  60 

Gly Ile Asp Val His Val Leu Gly Asp Ile Asn His Ile Ala Asn Ile 
65                  70                  75                  80 

Asp Leu Ile Phe Pro Asp Thr Ile Leu Thr Ala Asn Tyr Asn Gln His 
                85                  90                  95 

Gly Gln Ala Thr Thr Leu Thr Val Gln Ala Pro Ala Ser Ser Pro Ala 
            100                 105                 110 

Ser Val Ser His Val Pro Ser Ser Glu Pro Leu Pro Gln Ala Ser Ala 
        115                 120                 125 

Thr Ser Gln Pro Thr Val Pro Met Ala Pro Ser Ala Thr Pro Leu Ala 
    130                 135                 140 

Ser Ala Lys Pro Asp Ser Ser Val Thr Ala Ser Ser Glu Leu Thr Ser 
145                 150                 155                 160 

Ser Thr Asn Asp Val Ser Thr Glu Ser Ser Ser Glu Ser Gln Lys Gln 
                165                 170                 175 

Pro Glu Val Pro Gln Glu Ala Val Pro Thr Pro Lys Ala Ala Glu Thr 
            180                 185                 190 

Thr Glu Val Glu Pro Lys Thr Asp Ile Ser Glu Asp Pro Thr Ser Ala 
        195                 200                 205 

Asn Arg Pro Val Pro Asn Glu Ser Ala Ser Glu Glu Val Ser Ser Ala 
    210                 215                 220 

Ala Pro Ala Gln Ala Pro Ala Glu Lys Glu Glu Thr Ser Ala Pro Ala 
225                 230                 235                 240 

Ala Gln Lys Ala Val Ala Asp Thr Thr Ser Val Ala Thr Ser Asn Gly 
                245                 250                 255 

Leu Ser Tyr Ala Pro Asn His Ala Tyr Asn Pro Met Asn Ala Gly Leu 
            260                 265                 270 

Gln Pro Gln Thr Ala Ala Phe Lys Glu Glu Val Ala Ser Ala Phe Gly 
        275                 280                 285 

Ile Thr Ser Phe Ser Gly Tyr Arg Pro Gly Asp Pro Gly Asp His Gly 
    290                 295                 300 

Lys Gly Leu Ala Ile Asp Phe Met Val Pro Glu Asn Ser Ala Leu Gly 
305                 310                 315                 320 

Asp Gln Val Ala Gln Tyr Ala Ile Asp His Met Ala Glu Arg Gly Ile 
                325                 330                 335 

Ser Tyr Val Ile Trp Lys Gln Arg Phe Tyr Ala Pro Phe Ala Ser Ile 
            340                 345                 350 

Tyr Gly Pro Ala Tyr Thr Trp Asn Pro Met Pro Asp Arg Gly Ser Ile 
        355                 360                 365 

Thr Glu Asn His Tyr Asp His Val His Val Ser Phe Asn Ala 
    370                 375                 380 

 
           
             9  
             1095  
             DNA  
             S. pyogenes  
           
            9 

caagagtgga caccacgatc ggttacagaa atcaagtctg aactcgtcct agttgataat     60 

gtttttactt atactgtaaa atacggtgac actttaagca caattgctga agcaatggga    120 

attgatgtgc atgtcttagg agatattaat catattgcta atattgactt aatttttcca    180 

gacacgatcc taacagccaa ctacaaccaa cacggtcagg caacgacttt gacggttcaa    240 

gcgcctgctt ctagtccagc tagcgttagt catgtaccta gcagtgagcc attaccccaa    300 

gcatctgcca cctctcaatc gactgttcct atggcaccat ctgcgacacc atctgatgtc    360 

ccaacgacac cattcgcatc tgcaaagcca gatagttctg tgacagcgtc atctgagctc    420 

acatcgtcaa cgaatgatgt ttcgactgag ttgtctagcg aatcacaaaa gcagccagaa    480 

gtaccacaag aagcagttcc aactcctaaa gcagctgaaa cgactgaagt cgaacctaag    540 

acagacatct cagaggattc aacttcagct aataggcctg tacctaacga gagtgcttca    600 

gaagaagttt cttctgcggc cccagcacaa gccccagcag aaaaagaaga aacctctgcg    660 

ccagcagcac aaaaagctgt agctgacacc acaagtgttg caacctcaaa tggcctttct    720 

tacgctccaa accatgccta caatccaatg aatgcagggc ttcaaccaca aacagcagcc    780 

ttcaaagaag aagtggcttc tgcctttggt attacgtcat ttagtggtta ccgtccaggt    840 

gatccaggag atcatggtaa aggtttggcc attgatttta tggtgcctga aaattctgct    900 

cttggtgatc aagttgctca atatgccatt gaccatatgg cagagcgtgg tatttcatac    960 

gttatttgga aacagcgatt ctatgcgcca tttgcaagta tttacggacc agcctacaca   1020 

tggaacccca tgccagatcg cggcagtatt acagaaaacc attatgatca tgttcatgtc   1080 

tcctttaatg cttaa                                                    1095 

 
           
             10  
             364  
             PRT  
             S. pyogenes  
           
            10 

Gln Glu Trp Thr Pro Arg Ser Val Thr Glu Ile Lys Ser Glu Leu Val 
 1               5                  10                  15 

Leu Val Asp Asn Val Phe Thr Tyr Thr Val Lys Tyr Gly Asp Thr Leu 
            20                  25                  30 

Ser Thr Ile Ala Glu Ala Met Gly Ile Asp Val His Val Leu Gly Asp 
        35                  40                  45 

Ile Asn His Ile Ala Asn Ile Asp Leu Ile Phe Pro Asp Thr Ile Leu 
    50                  55                  60 

Thr Ala Asn Tyr Asn Gln His Gly Gln Ala Thr Thr Leu Thr Val Gln 
65                  70                  75                  80 

Ala Pro Ala Ser Ser Pro Ala Ser Val Ser His Val Pro Ser Ser Glu 
                85                  90                  95 

Pro Leu Pro Gln Ala Ser Ala Thr Ser Gln Ser Thr Val Pro Met Ala 
            100                 105                 110 

Pro Ser Ala Thr Pro Ser Asp Val Pro Thr Thr Pro Phe Ala Ser Ala 
        115                 120                 125 

Lys Pro Asp Ser Ser Val Thr Ala Ser Ser Glu Leu Thr Ser Ser Thr 
    130                 135                 140 

Asn Asp Val Ser Thr Glu Leu Ser Ser Glu Ser Gln Lys Gln Pro Glu 
145                 150                 155                 160 

Val Pro Gln Glu Ala Val Pro Thr Pro Lys Ala Ala Glu Thr Thr Glu 
                165                 170                 175 

Val Glu Pro Lys Thr Asp Ile Ser Glu Asp Ser Thr Ser Ala Asn Arg 
            180                 185                 190 

Pro Val Pro Asn Glu Ser Ala Ser Glu Glu Val Ser Ser Ala Ala Pro 
        195                 200                 205 

Ala Gln Ala Pro Ala Glu Lys Glu Glu Thr Ser Ala Pro Ala Ala Gln 
    210                 215                 220 

Lys Ala Val Ala Asp Thr Thr Ser Val Ala Thr Ser Asn Gly Leu Ser 
225                 230                 235                 240 

Tyr Ala Pro Asn His Ala Tyr Asn Pro Met Asn Ala Gly Leu Gln Pro 
                245                 250                 255 

Gln Thr Ala Ala Phe Lys Glu Glu Val Ala Ser Ala Phe Gly Ile Thr 
            260                 265                 270 

Ser Phe Ser Gly Tyr Arg Pro Gly Asp Pro Gly Asp His Gly Lys Gly 
        275                 280                 285 

Leu Ala Ile Asp Phe Met Val Pro Glu Asn Ser Ala Leu Gly Asp Gln 
    290                 295                 300 

Val Ala Gln Tyr Ala Ile Asp His Met Ala Glu Arg Gly Ile Ser Tyr 
305                 310                 315                 320 

Val Ile Trp Lys Gln Arg Phe Tyr Ala Pro Phe Ala Ser Ile Tyr Gly 
                325                 330                 335 

Pro Ala Tyr Thr Trp Asn Pro Met Pro Asp Arg Gly Ser Ile Thr Glu 
            340                 345                 350 

Asn His Tyr Asp His Val His Val Ser Phe Asn Ala 
        355                 360 

 
           
             11  
             1106  
             DNA  
             S. pyogenes  
           
            11 

caagagtgga caccacgatc ggttacagaa atcaagtctg aactcgtcct agttgataat     60 

gtttttactt atatagtaaa atacggtgac actttaagca caattgctga agcaatgggg    120 

attgatgtgc atgtcttagg agatattaat catattgcta atattgactt aatttttcca    180 

gacacgatcc taacagcaaa ctacaaccaa cacggtcagg caacgacttt gacggttcaa    240 

gcacctgctt ctagtccatc tagcgttagt catgtaccta gcagtgagcc attaccccaa    300 

gcatctgcca cctctcaacc gactgttcct atggcaccat ctgcgacacc atctgatgtc    360 

ccaacgacac cattcgcatc tgcaaagcca gatagttctg tgacagcgtc atctgagctc    420 

acatcgtcaa cgaatgatgt ttcgactgag ttgtctagcg aatcacaaaa gcagccagaa    480 

gtaccacaag aagcagttcc aactcctaaa gcagctgaac cgactgaagt cgaacctaag    540 

acagacatct cagaagaccc aacttcagct aataggcctg acctaacgag agtgcttcag    600 

aagaagcttc ttctgcggcc ccagcacaag ctccagcaga aaaagaagaa acctctcaga    660 

tgttaactgc gccagcagca caaaaagctg tagctgacac cacaagtgtt gcaacctcaa    720 

acggcctttc ttacgctcca aaccatgcct acaatccaat gaatgcaggg cttcaaccac    780 

aaacagcagc cttcaaagaa gaagtggctt ctgcctttgg tattacgtca tttagtggtt    840 

accgtccagg agatccagga gatcatggta aaggattagc cattgacttt atggtaccgg    900 

ttagctctac gcttggtgat caagttgctc aatatgccat tgaccatatg gcagagcgtg    960 

gtatttcata cgttatttgg aaacagcgat tctatgcgcc atttgcaagt atttacggac   1020 

cagcctacac atggaacccc atgccagatc gcggcagtat tacagaaaac cattatgatc   1080 

atgttcatgt ctcctttaat gcttaa                                        1106 

 
           
             12  
             368  
             PRT  
             S. pyogenes  
           
            12 

Gln Glu Trp Thr Pro Arg Ser Val Thr Glu Ile Lys Ser Glu Leu Val 
 1               5                  10                  15 

Leu Val Asp Asn Val Phe Thr Tyr Ile Val Lys Tyr Gly Asp Thr Leu 
            20                  25                  30 

Ser Thr Ile Ala Glu Ala Met Gly Ile Asp Val His Val Leu Gly Asp 
        35                  40                  45 

Ile Asn His Ile Ala Asn Ile Asp Leu Ile Phe Pro Asp Thr Ile Leu 
    50                  55                  60 

Thr Ala Asn Tyr Asn Gln His Gly Gln Ala Thr Thr Leu Thr Val Gln 
65                  70                  75                  80 

Ala Pro Ala Ser Ser Pro Ser Ser Val Ser His Val Pro Ser Ser Glu 
                85                  90                  95 

Pro Leu Pro Gln Ala Ser Ala Thr Ser Gln Pro Thr Val Pro Met Ala 
            100                 105                 110 

Pro Ser Ala Thr Pro Ser Asp Val Pro Thr Thr Pro Phe Ala Ser Ala 
        115                 120                 125 

Lys Pro Asp Ser Ser Val Thr Ala Ser Ser Glu Leu Thr Ser Ser Thr 
    130                 135                 140 

Asn Asp Val Ser Thr Glu Leu Ser Ser Glu Ser Gln Lys Gln Pro Glu 
145                 150                 155                 160 

Val Pro Gln Glu Ala Val Pro Thr Pro Lys Ala Ala Glu Pro Thr Glu 
                165                 170                 175 

Val Glu Pro Lys Thr Asp Ile Ser Glu Asp Pro Thr Ser Ala Asn Arg 
            180                 185                 190 

Pro Val Pro Asn Glu Ser Ala Ser Glu Glu Ala Ser Ser Ala Ala Pro 
        195                 200                 205 

Ala Gln Ala Pro Ala Glu Lys Glu Glu Thr Ser Gln Met Leu Thr Ala 
    210                 215                 220 

Pro Ala Ala Gln Lys Ala Val Ala Asp Thr Thr Ser Val Ala Thr Ser 
225                 230                 235                 240 

Asn Gly Leu Ser Tyr Ala Pro Asn His Ala Tyr Asn Pro Met Asn Ala 
                245                 250                 255 

Gly Leu Gln Pro Gln Thr Ala Ala Phe Lys Glu Glu Val Ala Ser Ala 
            260                 265                 270 

Phe Gly Ile Thr Ser Phe Ser Gly Tyr Arg Pro Gly Asp Pro Gly Asp 
        275                 280                 285 

His Gly Lys Gly Leu Ala Ile Asp Phe Met Val Pro Val Ser Ser Thr 
    290                 295                 300 

Leu Gly Asp Gln Val Ala Gln Tyr Ala Ile Asp His Met Ala Glu Arg 
305                 310                 315                 320 

Gly Ile Ser Tyr Val Ile Trp Lys Gln Arg Phe Tyr Ala Pro Phe Ala 
                325                 330                 335 

Ser Ile Tyr Gly Pro Ala Tyr Thr Trp Asn Pro Met Pro Asp Arg Gly 
            340                 345                 350 

Ser Ile Thr Glu Asn His Tyr Asp His Val His Val Ser Phe Asn Ala 
        355                 360                 365 

 
           
             13  
             1095  
             DNA  
             S. pyogenes  
           
            13 

caagagtgga caccacgatc ggttacagaa atcaagtctg aactcgtcct agttgataat     60 

gtttttactt atactgtaaa atacggtgac actttaagca caattgctga agcaatgggg    120 

attgatgtgc atgtcttagg agatattaat catattgcta atattgacct aatttttcca    180 

gacacgatcc taacagcaaa ctacaatcaa cacggtcagg caacgaattt gacggttcaa    240 

gcacctgctt ctagtccagc tagcgttagt catgtaccta gcagtgagcc attaccccaa    300 

gcatctgcca cctctcaacc gactgttcct atggcaccac ctgcgacacc atctgatgtc    360 

ccaacgacac cattcgcatc tgcaaagcca gatagttctg tgacagcgtc atctgagctc    420 

acatcgtcaa cgaatgatgt ttcgactgag ttgtctagcg aatcacaaaa gcagccagaa    480 

gtaccacaag aagcagttcc aactcctaaa gcagctgaaa cgactgaagt cgaacctaag    540 

acagacatct cagaagcccc aacttcagct aataggcctg tacctaacga gagtgcttca    600 

gaagaagttt cttctgcggc cccagcacaa gccccagcag aaaaagaaga aacctctgcg    660 

ccagcagcac aaaaagctgt agctgacacc acaagtgttg caacctcaaa tggcctttct    720 

tacgctccaa accatgccta caatccaatg aatgcagggc ttcaaccaca aacagcagcc    780 

ttcaaagaag aagtggcttc tgcctttggt attacgtcat ttagtggtta ccgtccaggt    840 

gatccaggag atcatggtaa aggtttggcc attgatttta tggtgcctga aaattctgct    900 

cttggtgatc aagttgctca atatgccatt gaccatatgg cagagcgtgg tatttcatac    960 

gttatttgga aacagcgatt ctatgcgcca tttgcaagta tttacggacc agcctacaca   1020 

tggaacccca tgccagatcg cggcagtatt acagaaaacc attatgatca tgttcatgtc   1080 

tcctttaatg cttaa                                                    1095 

 
           
             14  
             364  
             PRT  
             S. pyogenes  
           
            14 

Gln Glu Trp Thr Pro Arg Ser Val Thr Glu Ile Lys Ser Glu Leu Val 
 1               5                  10                  15 

Leu Val Asp Asn Val Phe Thr Tyr Thr Val Lys Tyr Gly Asp Thr Leu 
            20                  25                  30 

Ser Thr Ile Ala Glu Ala Met Gly Ile Asp Val His Val Leu Gly Asp 
        35                  40                  45 

Ile Asn His Ile Ala Asn Ile Asp Leu Ile Phe Pro Asp Thr Ile Leu 
    50                  55                  60 

Thr Ala Asn Tyr Asn Gln His Gly Gln Ala Thr Asn Leu Thr Val Gln 
65                  70                  75                  80 

Ala Pro Ala Ser Ser Pro Ala Ser Val Ser His Val Pro Ser Ser Glu 
                85                  90                  95 

Pro Leu Pro Gln Ala Ser Ala Thr Ser Gln Pro Thr Val Pro Met Ala 
            100                 105                 110 

Pro Pro Ala Thr Pro Ser Asp Val Pro Thr Thr Pro Phe Ala Ser Ala 
        115                 120                 125 

Lys Pro Asp Ser Ser Val Thr Ala Ser Ser Glu Leu Thr Ser Ser Thr 
    130                 135                 140 

Asn Asp Val Ser Thr Glu Leu Ser Ser Glu Ser Gln Lys Gln Pro Glu 
145                 150                 155                 160 

Val Pro Gln Glu Ala Val Pro Thr Pro Lys Ala Ala Glu Thr Thr Glu 
                165                 170                 175 

Val Glu Pro Lys Thr Asp Ile Ser Glu Ala Pro Thr Ser Ala Asn Arg 
            180                 185                 190 

Pro Val Pro Asn Glu Ser Ala Ser Glu Glu Val Ser Ser Ala Ala Pro 
        195                 200                 205 

Ala Gln Ala Pro Ala Glu Lys Glu Glu Thr Ser Ala Pro Ala Ala Gln 
    210                 215                 220 

Lys Ala Val Ala Asp Thr Thr Ser Val Ala Thr Ser Asn Gly Leu Ser 
225                 230                 235                 240 

Tyr Ala Pro Asn His Ala Tyr Asn Pro Met Asn Ala Gly Leu Gln Pro 
                245                 250                 255 

Gln Thr Ala Ala Phe Lys Glu Glu Val Ala Ser Ala Phe Gly Ile Thr 
            260                 265                 270 

Ser Phe Ser Gly Tyr Arg Pro Gly Asp Pro Gly Asp His Gly Lys Gly 
        275                 280                 285 

Leu Ala Ile Asp Phe Met Val Pro Glu Asn Ser Ala Leu Gly Asp Gln 
    290                 295                 300 

Val Ala Gln Tyr Ala Ile Asp His Met Ala Glu Arg Gly Ile Ser Tyr 
305                 310                 315                 320 

Val Ile Trp Lys Gln Arg Phe Tyr Ala Pro Phe Ala Ser Ile Tyr Gly 
                325                 330                 335 

Pro Ala Tyr Thr Trp Asn Pro Met Pro Asp Arg Gly Ser Ile Thr Glu 
            340                 345                 350 

Asn His Tyr Asp His Val His Val Ser Phe Asn Ala 
        355                 360 

 
           
             15  
             1074  
             DNA  
             S. pyogenes  
           
            15 

caagagtgga caccacgatc ggttacagaa atcaagtctg aactcgtcct agttgataat     60 

gtttttactt atacagtaaa atacggtgac actttaagca caattgctga agcaatgggg    120 

attgatgtgc atgtcttagg agatattaat catattgcta atattgactt aatttttcca    180 

gacacgatcc taacagcaaa ctacaatcaa cacggtcagg caacgacttt gacggttcaa    240 

gcacctgctt ctagtccagc tagcgttagt catgtaccta gcagtgagcc attaccccaa    300 

gcatctgcca cctctcaacc gactgttcct atggcaccat ctgcgacacc attagcatct    360 

gcaaagccag atagttctgt gacagcgtca tctgagctca catcgtcaac gaatgatgtt    420 

tcgactgagt cgtctagcga atcacaaaag cagccagaag taccacaaga agcagttcca    480 

actcctaaag cagctgaaac gactgaagtc gaacctaaga cagacatctc agaagaccca    540 

acttcagcta ataggcctgt acctaacgag agtgcttcag aagaagtttc ttctgcggcc    600 

ccagcacaag ccccagcaga aaaagaagaa acctctgcgc cagcagcaca aaaagctgta    660 

gctgacacca caagtgttgc aacctcaaac ggcctttctt acgctccaaa ccatgcctac    720 

aatccaatga atgcagggct tcaaccacaa acagcagcct tcaaagaaga agtggcttct    780 

gcctttggta ttacgtcatt tagtggttac cgtccaggtg acccaggaga tcatggtaaa    840 

ggtttggcca ttgattttat ggtgcctgaa aattctgctc ttggtgatca agttgctcaa    900 

tatgccattg accatatggc agagcgtggt atttcatacg ttatttggaa acagcgattc    960 

tatgcgccat ttgcaagtat ttacggacca gcttacacat ggaaccccat gccagatcgc   1020 

ggcagtatta cagaaaacca ttatgatcat gttcatgtct cctttaatgc ttaa         1074 

 
           
             16  
             357  
             PRT  
             S. pyogenes  
           
            16 

Gln Glu Trp Thr Pro Arg Ser Val Thr Glu Ile Lys Ser Glu Leu Val 
 1               5                  10                  15 

Leu Val Asp Asn Val Phe Thr Tyr Thr Val Lys Tyr Gly Asp Thr Leu 
            20                  25                  30 

Ser Thr Ile Ala Glu Ala Met Gly Ile Asp Val His Val Leu Gly Asp 
        35                  40                  45 

Ile Asn His Ile Ala Asn Ile Asp Leu Ile Phe Pro Asp Thr Ile Leu 
    50                  55                  60 

Thr Ala Asn Tyr Asn Gln His Gly Gln Ala Thr Thr Leu Thr Val Gln 
65                  70                  75                  80 

Ala Pro Ala Ser Ser Pro Ala Ser Val Ser His Val Pro Ser Ser Glu 
                85                  90                  95 

Pro Leu Pro Gln Ala Ser Ala Thr Ser Gln Pro Thr Val Pro Met Ala 
            100                 105                 110 

Pro Ser Ala Thr Pro Leu Ala Ser Ala Lys Pro Asp Ser Ser Val Thr 
        115                 120                 125 

Ala Ser Ser Glu Leu Thr Ser Ser Thr Asn Asp Val Ser Thr Glu Ser 
    130                 135                 140 

Ser Ser Glu Ser Gln Lys Gln Pro Glu Val Pro Gln Glu Ala Val Pro 
145                 150                 155                 160 

Thr Pro Lys Ala Ala Glu Thr Thr Glu Val Glu Pro Lys Thr Asp Ile 
                165                 170                 175 

Ser Glu Asp Pro Thr Ser Ala Asn Arg Pro Val Pro Asn Glu Ser Ala 
            180                 185                 190 

Ser Glu Glu Val Ser Ser Ala Ala Pro Ala Gln Ala Pro Ala Glu Lys 
        195                 200                 205 

Glu Glu Thr Ser Ala Pro Ala Ala Gln Lys Ala Val Ala Asp Thr Thr 
    210                 215                 220 

Ser Val Ala Thr Ser Asn Gly Leu Ser Tyr Ala Pro Asn His Ala Tyr 
225                 230                 235                 240 

Asn Pro Met Asn Ala Gly Leu Gln Pro Gln Thr Ala Ala Phe Lys Glu 
                245                 250                 255 

Glu Val Ala Ser Ala Phe Gly Ile Thr Ser Phe Ser Gly Tyr Arg Pro 
            260                 265                 270 

Gly Asp Pro Gly Asp His Gly Lys Gly Leu Ala Ile Asp Phe Met Val 
        275                 280                 285 

Pro Glu Asn Ser Ala Leu Gly Asp Gln Val Ala Gln Tyr Ala Ile Asp 
    290                 295                 300 

His Met Ala Glu Arg Gly Ile Ser Tyr Val Ile Trp Lys Gln Arg Phe 
305                 310                 315                 320 

Tyr Ala Pro Phe Ala Ser Ile Tyr Gly Pro Ala Tyr Thr Trp Asn Pro 
                325                 330                 335 

Met Pro Asp Arg Gly Ser Ile Thr Glu Asn His Tyr Asp His Val His 
            340                 345                 350 

Val Ser Phe Asn Ala 
        355 

 
           
             17  
             1113  
             DNA  
             S. pneumonia  
           
            17 

atgaagaaaa gaatgttatt agcgtcaaca gtagccttgt catttgcccc agtattggca     60 

actcaagcag aagaagttct ttggactgca cgtagtgttg agcaaatcca aaacgatttg    120 

actaaaacgg acaacaaaac aagttatacc gtacagtatg gtgatacttt gagcaccatt    180 

gcagaagcct tgggtgtaga tgtcacagtg cttgcgaatc tgaacaaaat cactaatatg    240 

gacttgattt tcccagaaac tgttttgaca acgactgtca atgaagcaga agaagtaaca    300 

gaagttgaaa tccaaacacc tcaagcagac tctagtgaag aagtgacaac tgcgacagca    360 

gatttgacca ctaatcaagt gaccgttgat gatcaaactg ttcaggttgc agacctttct    420 

caaccaattg cagaagttac aaagacagtg attgcttctg aagaagtggc accatctacg    480 

ggcacttctg tcccagagga gcaaacgacc gaaacaactc gcccagttga agaagcaact    540 

cctcaggaaa cgactccagc tgagaagcag gaaacacaag caagccctca agctgcatca    600 

gcagtggaag taactacaac aagttcagaa gcaaaagaag tagcatcatc aaatggagct    660 

acagcagcag tttctactta tcaaccagaa gagacgaaaa taatttcaac aacttacgag    720 

gctccagctg cgcccgatta tgctggactt gcagtagcaa aatctgaaaa tgcaggtctt    780 

caaccacaaa cagctgcctt taaagaagaa attgctaact tgtttggcat tacatccttt    840 

agtggttatc gtccaggaga cagtggagat cacggaaaag gtttggctat cgactttatg    900 

gtaccagaac gttcagaatt aggggataag attgcggaat atgctattca aaatatggcc    960 

agccgtggca ttagttacat catctggaaa caacgtttct atgctccatt cgatagcaaa   1020 

tatgggccag ctaacacttg gaacccaatg ccagaccgtg gtagtgtgac agaaaatcac   1080 

tatgatcacg ttcacgtttc aatgaatgga taa                                1113 

 
           
             18  
             370  
             PRT  
             S. pneumonia  
           
            18 

Met Lys Lys Arg Met Leu Leu Ala Ser Thr Val Ala Leu Ser Phe Ala 
 1               5                  10                  15 

Pro Val Leu Ala Thr Gln Ala Glu Glu Val Leu Trp Thr Ala Arg Ser 
            20                  25                  30 

Val Glu Gln Ile Gln Asn Asp Leu Thr Lys Thr Asp Asn Lys Thr Ser 
        35                  40                  45 

Tyr Thr Val Gln Tyr Gly Asp Thr Leu Ser Thr Ile Ala Glu Ala Leu 
    50                  55                  60 

Gly Val Asp Val Thr Val Leu Ala Asn Leu Asn Lys Ile Thr Asn Met 
65                  70                  75                  80 

Asp Leu Ile Phe Pro Glu Thr Val Leu Thr Thr Thr Val Asn Glu Ala 
                85                  90                  95 

Glu Glu Val Thr Glu Val Glu Ile Gln Thr Pro Gln Ala Asp Ser Ser 
            100                 105                 110 

Glu Glu Val Thr Thr Ala Thr Ala Asp Leu Thr Thr Asn Gln Val Thr 
        115                 120                 125 

Val Asp Asp Gln Thr Val Gln Val Ala Asp Leu Ser Gln Pro Ile Ala 
    130                 135                 140 

Glu Val Thr Lys Thr Val Ile Ala Ser Glu Glu Val Ala Pro Ser Thr 
145                 150                 155                 160 

Gly Thr Ser Val Pro Glu Glu Gln Thr Thr Glu Thr Thr Arg Pro Val 
                165                 170                 175 

Glu Glu Ala Thr Pro Gln Glu Thr Thr Pro Ala Glu Lys Gln Glu Thr 
            180                 185                 190 

Gln Ala Ser Pro Gln Ala Ala Ser Ala Val Glu Val Thr Thr Thr Ser 
        195                 200                 205 

Ser Glu Ala Lys Glu Val Ala Ser Ser Asn Gly Ala Thr Ala Ala Val 
    210                 215                 220 

Ser Thr Tyr Gln Pro Glu Glu Thr Lys Ile Ile Ser Thr Thr Tyr Glu 
225                 230                 235                 240 

Ala Pro Ala Ala Pro Asp Tyr Ala Gly Leu Ala Val Ala Lys Ser Glu 
                245                 250                 255 

Asn Ala Gly Leu Gln Pro Gln Thr Ala Ala Phe Lys Glu Glu Ile Ala 
            260                 265                 270 

Asn Leu Phe Gly Ile Thr Ser Phe Ser Gly Tyr Arg Pro Gly Asp Ser 
        275                 280                 285 

Gly Asp His Gly Lys Gly Leu Ala Ile Asp Phe Met Val Pro Glu Arg 
    290                 295                 300 

Ser Glu Leu Gly Asp Lys Ile Ala Glu Tyr Ala Ile Gln Asn Met Ala 
305                 310                 315                 320 

Ser Arg Gly Ile Ser Tyr Ile Ile Trp Lys Gln Arg Phe Tyr Ala Pro 
                325                 330                 335 

Phe Asp Ser Lys Tyr Gly Pro Ala Asn Thr Trp Asn Pro Met Pro Asp 
            340                 345                 350 

Arg Gly Ser Val Thr Glu Asn His Tyr Asp His Val His Val Ser Met 
        355                 360                 365 

Asn Gly 
    370 

 
           
             19  
             1183  
             DNA  
             S. pyogenes  
             
               misc_difference  
               (428)...(448)  
               nnnnnnnnnnnnnnnnnnnnn can be 
      ctgatgtccaacgacaccat or absent  
             
           
            19 

atgattatta ctaaaaagag yttatttgtg acaagtgtcg ctttgtcgtt agyacctttg     60 

gcgacagcrc aggcacaaga gtggacacca cgatcggtta casaaatcaa gtctgaactc    120 

gtcctagttg ataatgtttt tacttatayw gtaaaatacg gtgacacttt aagcacaatt    180 

gctgaagcaa tgggrattga tgtgcatgtc ttaggagata ttaatcatat tgctaatatt    240 

gacytaattt ttccagacac gatcctaaca gcmaactaca aycaacacgg tcaggcaacg    300 

amtttgacgg ttcaagcrcc tgcttctagt ccakctagcg ttagtcatgt acctagcagt    360 

gagccattac cccaagcatc tgccacctct caaycgactr ttcctatggc accayctgcg    420 

acaccatnnn nnnnnnnnnn nnnnnnnntm gcatctgcaa agccagatag ttytgtgaca    480 

gcgtcatctg agctcacatc rtcaacgaat gatgtttcga ctgagtygtc tagcgaatca    540 

caaaagcagc cagaagtacc acaagaagca gwwccaactc ctaaagcagc tgaamssact    600 

gaagtcgaac ctaagacaga catctcagar gmyycaactt cagctaatag gcctgtacct    660 

aacgrragtg cttcagaaga agyttcttct gcggccccag cacaagcycc agcagaaaaa    720 

gaagaaacct ctnnnnnnnn nnnngcgcca gcagcacaaa aagctgtagc tgacaccaca    780 

agtgttgcaa cctcaaaygg cctttcttac gctccaaacc atgcctacaa tccaatgaat    840 

gcagggcttc aaccacaaac agcagccttc aaagaagaag tgncttctgc ctttggtatt    900 

acgtcattta gtggttaccg tccaggwgay ccaggagatc atnggtaaag gwttrgccat    960 

tgaytttatg gtrcckgwwa rytctrckct tggtgatcaa gttgctcaat atgccattga   1020 

ccatatggca gassgtggta tttcatacgt tatttggaaa cagcgattct atgcgccatt   1080 

tgcaagtatt tacggaccag cytacacatg gaaccccatg ccagatcgcg gcagtattac   1140 

agwwwwccat tatgatcatg ttcatgtctc ctttaatgct taa                     1183 

 
           
             20  
             393  
             PRT  
             s. pyogenes  
             
               VARIANT  
               (18)...(18)  
               Xaa = Ala or Val  
             
           
            20 

Met Ile Ile Thr Lys Lys Ser Leu Phe Val Thr Ser Val Ala Leu Ser 
 1               5                  10                  15 

Leu Xaa Pro Leu Ala Thr Ala Gln Ala Gln Glu Trp Thr Pro Arg Ser 
            20                  25                  30 

Val Thr Glx Ile Lys Ser Glu Leu Val Leu Val Asp Asn Val Phe Thr 
        35                  40                  45 

Tyr Xaa Val Lys Tyr Gly Asp Thr Leu Ser Thr Ile Ala Glu Ala Met 
    50                  55                  60 

Gly Ile Asp Val His Val Leu Gly Asp Ile Asn His Ile Ala Asn Ile 
65                  70                  75                  80 

Asp Leu Ile Phe Pro Asp Thr Ile Leu Thr Ala Asn Tyr Asn Gln His 
                85                  90                  95 

Gly Gln Ala Thr Xaa Leu Thr Val Gln Ala Pro Ala Ser Ser Pro Xaa 
            100                 105                 110 

Ser Val Ser His Val Pro Ser Ser Glu Pro Leu Pro Gln Ala Ser Ala 
        115                 120                 125 

Thr Ser Gln Xaa Thr Xaa Pro Met Ala Pro Xaa Ala Thr Pro Xaa Xaa 
    130                 135                 140 

Xaa Xaa Xaa Xaa Xaa Xaa Ala Ser Ala Lys Pro Asp Ser Xaa Val Thr 
145                 150                 155                 160 

Ala Ser Ser Glu Leu Thr Ser Ser Thr Asn Asp Val Ser Thr Glu Xaa 
                165                 170                 175 

Ser Ser Glu Ser Gln Lys Gln Pro Glu Val Pro Gln Glu Ala Xaa Pro 
            180                 185                 190 

Thr Pro Lys Ala Ala Glu Xaa Thr Glu Val Glu Pro Lys Thr Asp Ile 
        195                 200                 205 

Ser Glu Xaa Xaa Thr Ser Ala Asn Arg Pro Val Pro Asn Xaa Ser Ala 
    210                 215                 220 

Ser Glu Glu Xaa Ser Ser Ala Ala Pro Ala Gln Ala Pro Ala Glu Lys 
225                 230                 235                 240 

Glu Xaa Xaa Xaa Xaa Xaa Xaa Xaa Ala Pro Ala Ala Gln Lys Ala Val 
                245                 250                 255 

Ala Asp Thr Thr Ser Val Ala Thr Ser Asn Gly Leu Ser Tyr Ala Pro 
            260                 265                 270 

Asn His Ala Tyr Asn Pro Met Asn Ala Gly Leu Gln Pro Gln Thr Ala 
        275                 280                 285 

Ala Phe Lys Glu Glu Val Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 
    290                 295                 300 

Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Gly Lys Gly Leu Ala Ile 
305                 310                 315                 320 

Asp Phe Met Val Pro Xaa Xaa Ser Xaa Leu Gly Asp Gln Val Ala Gln 
                325                 330                 335 

Tyr Ala Ile Asp His Met Ala Xaa Xaa Gly Ile Ser Tyr Val Ile Trp 
            340                 345                 350 

Lys Gln Arg Phe Tyr Ala Pro Phe Ala Ser Ile Tyr Gly Pro Ala Tyr 
        355                 360                 365 

Thr Trp Asn Pro Met Pro Asp Arg Gly Ser Ile Thr Xaa Xaa His Tyr 
    370                 375                 380 

Asp His Val His Val Ser Phe Asn Ala 
385                 390 

 
           
             21  
             32  
             DNA  
             Artificial Sequence  
             
               DMAR16 Oligonucleotide  
             
           
            21 

caggccatgg agtggacacc acgatcggtt ac                                   32 

 
           
             22  
             37  
             DNA  
             Artificial Sequence  
             
               DMAR17 Oligonucleotide  
             
           
            22 

gccgctcgag agcattaaag gagacatgaa catgatc                              37 

 
           
             23  
             25  
             PRT  
             Artificial Sequence  
             
               Signal peptide predicted from analysis of SEQ 
      ID NO2  
             
           
            23 

Met Ile Ile Thr Lys Lys Ser Leu Phe Val Thr Ser Val Ala Leu Ser 
 1               5                  10                  15 

Leu Ala Pro Leu Ala Thr Ala Gln Ala 
            20                  25 

 
           
             24  
             5  
             PRT  
             Artificial Sequence  
             
               VARIANT  
               (3)...(3)  
               Xaa = Any Amino Acid  
             
           
            24 

Leu Pro Xaa Thr Gly 
 1               5 

 
           
             25  
             6  
             PRT  
             Artificial Sequence  
             
               IgA binding motif  
             
           
            25 

Met Leu Lys Lys Ile Glu 
 1               5 

 
           
             26  
             28  
             DNA  
             Artificial Sequence  
             
               DMAR69 oligonucleotide  
             
           
            26 

ctgggaagat tatctagcac attaatac                                        28 

 
           
             27  
             25  
             DNA  
             Artificial Sequence  
             
               DMAR72 oligonucleotide  
             
           
            27 

cataacgtta aaactgtcta aaggg                                           25 

 
           
             28  
             34  
             DNA  
             Artificial Sequence  
             
               DMAR24 oligonucleotide  
             
           
            28 

tacccggatc cccaagagtg gacaccacga tcgg                                 34 

 
           
             29  
             36  
             DNA  
             Artificial Sequence  
             
               DMAR25 oligonucleotide  
             
           
            29 

gcgctcgtcg acgcgtatct cagcctctta tagggc                               36