AU2004201404B2 - Genes and proteins, and their use - Google Patents

Description

Regulation 3.2

AUSTRALIA

Patents Act 1990 COMPLETE SPECIFICATION STANDARD PATENT

APPLICANT:

Invention Title: Microscience Limited GENES AND PROTEINS, AND THEIR USE The following statement is a full description of this invention, including the best method of performing it known to me: GENES AND PROTEINS. AND THEIR USE Field of the Invention This invention relates to the identification c bacterial genes and proteins, and their use. MoZ particularly, it relates to their use in therapy, fc immunisation and in screening for drugs.

Background to the Invention Group B Streptococcus (GBS), also known a Streptococcus agalactlae, is the causative agent of variou conditions. In particular, GBS causes: Early onset neonatal Infection.

This infection usually begins In utero and cause severe septicaemia and pneumonia in infants, which i lethal if untreated and even with treatment is associate with a 10-20% mortality rate.

Late onset neonatal Infection.

This infection occurs in the period shortly afte birth until about 3 months of age. It causes septicaemia, which is complicated by meningitis in 90% c cases. Other focal infections also occur includi osteomyelitis, septic arthritis, abscesses an endopthalmitis.

Adult Infections.

These appear to be Increasingly common and occur mos frequently in women who have just delivered a baby, th elderly and the immunocompromised. They are characterise by septicaemia and focal infections includin osteomyelitis, septic arthritis, abscesses an endopthalmitis.

Urinary tract Infections.

GBS is a cause of urinary tract infections and i pregnancy accounts for about 10% of all infections.

Veterinary infections.

GBS causes chronic mastitis in cows. This, in turn leads to reduced milk production and is therefore o considerable economic importance.

GBS infections can be treated with antibiotic However, immunisation is preferable. It is therefoi desirable to develop an immunogen that could be used in therapeutically-effective vaccine.

Summary of the Invention The present invention is based on the identificatic of a series of genes in GBS, and also related organisms the products of which may be localised on the outer surfac of the organism and therefore may be used as a target fc immuno-therapy.

According to one aspect of the invention, a peptide i encoded by an operon including any of the genes identifie herein as phol-13, pho3-21, pho2-15, .pho3-18, pho3-22 pho3-3, pho3-17, pho2-2, phol-5, pho3-1, pho3-23, pho3-50 phol-14, pho2-10, pho3-14, pho3-24 and pho3-29, obtainabl from Group B Streptococcus, or a homologue or functiona fragment thereof. Such a peptide is suitable fo therapeutic use, e.g. when isolated.

The term "functional fragments" is used herein t define a part of the gene or peptide which retains thb activity of the whole gene or peptide. For example, functional fragment of the peptide may be used as ai antigenic determinant, useful in a vaccine or in the production of antibodies.

A gene fragment may be used to encode the activi peptide. Alternatively, the gene fragment may have utilit] in gene therapy, targetting the wild-type gene In vivo tc exert a therapeutic effect.

A peptide according to the present invention maj comprise any of the amino acid sequences identified hereir as SEQ ID NOS. 2, 4, 6, 8, 10, 11, 13, 15, 17, 19, 21, 23, 27, 29, 31, 33 and 35, or a functional fragment thereof.

Because of the extracellular or cell surface location, the peptides of the present invention may be suitable candidates for the production of therapeutically-effective vaccines against GBS. The term "therapeutically-effective" is intended to include the prophylactic effect of vaccine: or..example, a vaccine may comprise a peptide according the invention, or the means for its expression, for tl treatment of infection. The vaccine may be administered 1 females prior to or during pregnancy to protect mother as neonate against infection by GBS.

According to another aspect of the invention, t peptides or genes may be used for screening potentie antimicrobial drugs or for the detection of virulence.

A further aspect of this invention is the use of ar of the products identified herein, for the treatment prevention- of a condition associated with infection by Group B Streptococcal strain.

Although the protein has been described for use in' t treatment of patients, veterinary uses of the products C the invention are also considered to be within the scope c the present invention. In particular, the peptides or th vaccines may be used in the treatment of chronic mastitis especially in cows.

Description of the Invention The present invention is described with reference t Group B Streptococcal strain M732. However, all the GB strains and many other bacterial strains are likely t include related peptides or proteins having amino aci sequence homology with the peptide of M732. Organism likely to contain the peptides include, but are not limite to, S. pneumoniae, S. pyogenes, S. suis, S. milleri, Grou: C and Group G Streptococci and Enterococci. Vaccines t, each of these may be developed in the same way as describe for GBS.

Preferably, the peptides that may be useful for th production of vaccines have greater than 40% sequence similarity with the peptides identified herein. Mor preferably, the peptides have greater than 60% sequence similarity. Most preferably, the peptides have greates than 80% sequence similarity, e.g. 95% similarity.

a r5 WfI wfrJW II 4 Having characterised a gene according to t invention, it is possible to use the gene sequence establish homologies in other microorganisms. In this v it is possible to determine whether other microorganis have similar outer surface products. Sequence homologi may be established by searching in existing databases, e.

EMBL or Genbank.

Peptides or proteins according to the invention may purified and isolated by methods known in the art.

particular, having identified the gene sequence, it will possible to use recombinant techniques to express the gen in a suitable-host.- Active fragments and homologues can identified and may be useful in therapy. For example,' t peptides or their active fragments may be used as antigen determinants in a vaccine, to elicit an immune respons They may also be used in the preparation of antibodies, f' passive immunisation, or diagnostic applications. Suitab antibodies include monoclonal antibodies, or fragmenthereof, including single chain fv fragments. Methods f, the preparation of antibodies will be apparent to tho: skilled in the art.

The preparation of vaccines based on attenuat microorganisms is known to those skilled in the art Vaccine compositions can be formulated with suitab: carriers or adjuvants, e.g. alum, as necessary or desirec and used in therapy, to provide effective immunisati< against Group B Streptococci or other relat microorganisms. The preparation of vaccine. formulatioz will be apparent to the skilled person.

More generally, and as is well known to those skille in the art, a suitable amount of an active component of tt invention can be selected, for therapeutic use, as ce suitable carriers or excipients, and routes c administration. These factors will be chosen or determine according to known criteria such as the nature/severity c the condition to be treated, the type or health of tk subject etc.

The products of the present invention were identifit as follows: A partial gene library of GBS (strain M732 chromosomal DNA was prepared using the plasmid vectors pF phoAl, pFW-phoA2 and pFW-phoA3 (Podbielski, A. et al. 1996 Gene 177:137-147). These plasmids possess a constitutiv spectinomycin adenyltransferase antibiotic resistanc marker, which confers a high level of spectinomyci resistance and is therefore easily selected. Furthermore these vectors contain a truncated (leaderless) Escherichi coll phoA gene for alkaline phosphatase.. The three vector differ only with respect to the reading:frame in which th leaderless phoA gene exists, as compared to an, upstream in frame BamHI restriction enzyme site. .Because thi truncated X. coli phoA gene lacks the appropriate leade sequence for export of this enzyme across the bacteria membrane, extracellular alkaline phosphatase activity i absent when these plasmids are propagated in an col.

phoA mutant strain DH5). The chromogenic alkalin phosphatase substrate, XP (5-bromo-4-chloro-3-indolyl phosphate), does not enter intact bacterial cells an therefore only exported or surface associated alkalin, phosphatase activity can be detected. When exported o: surface associated alkaline phosphatase activity ii present, the chromogenic XP substrate is cleaved to yiel< a blue pigment and the corresponding bacterial colonies cal be identified by their blue colour.

Plasmid DNA was digested to completion with BamHI an< dephosphorylated using shrimp alkaline phosphatase. GBI genomic DNA was partially digested with Sau3AI, siz fractionated on a sucrose gradient and fragments <lkb ii size were ligated into the prepared pFW-phoA vectors. E.

coll strain DH5a was chosen as the cloning host since it lacks a functional phoA gene. Recombinant plasmids wer selected on Luria agar containing 100 pg/ml ol spectinomycin and 40 pg/ml of the chromogenic XP substrate.

E. coli transformants harbouring plasmids containing GBE rT. I IDWI V7J l I 6 insert DNA that complements the export signal sequence the leaderless phoA gene were identified by the blue colc of the colonies. Approximately 30000 different recombina plasmids containing GBS insert DNA were screened in th manner and 83 recombinant plasmids, which complemented t leaderless phoA, were chosen for further study.

From these experiments, several clones were select each containing a plasmid containing a gene (or pa.

thereof), which complemented the leaderless phoA.

Having identified the gene in each clone it is th.

possible to obtain the full-length gene sequence, follows. Using the identified and sequenced gene fragmeni oligonucleotide primers were designed for genomic DI sequencing. These primers were designed so as to sequent in an 'outwardO direction.from the obtained sequence. Ont read, the sequence obtained was checked to see if the and 3' termini of the gene had been reached. The present of these features was identified by checking agains homologous sequences, and for the 5' end the presence of a AUG start codon (or accepted equivalent) preceded by Shine-Dalgarno consensus sequence, and for the 3' end, tU presence of a translation termination (Stop) codon.

Upon identification of the full-length gene, primer were designed for amplification of full-length product Primers used included restriction enzyme recognition site (Ncol at the S'end and EcoO109I at the 3' end) to alla subsequent cloning of the product into the Lactococca expression system used.

PCR was carried out using the primers, and th products cloned into a pCR 2.1 cloning vector (I Vitrogen). Following confirmation of the presence of th cloned fragment, the DNA was excised using the restrictio enzymes NcoI and Eco0109I.

The vector into which this fragment was inserted wa; a modified version of pNZ8048 (Kuipers, O. P. et al. (1998 J. Biotech 64: 15-21). This vector, harbouring i lactococcal origin of replication, a chloramphenico resistance- marker, -an- inducible nisin promoter and multicloning site was altered by the replacement of th multicloning site with two 10X His tags, flanked on the most end with an Ncol site, split in the middle with multicloning site (including an EcoO109I site), and a Sto (termination) codon at the .3'end of the His tags.

The gene of interest was inserted so that a 10X Hi tag was in the 3' position relative to the coding region Following transformation of the recombinant plasmid int L.lactis (strain NZ9000 Kuipers, O. P. et al. (1998 supra), a 400 ml liquid culture was set up and translatio of the protein was. induced by the addition of nisin to th culture. After a 2 hour incubation, the cells wer harvested and lysed by bead beating. The resultant lysat was cleared by centrifugation, then passed over a meta affinity (Talon, Clonetech) column. The column was washe repeatedly before bound proteins were eluted wit Imidazole.

To identify fractions containing the His-tagge recombinant protein, an aliquot from each fraction wa analysed by SDS-PAGE, Western blotted and probed with anti His antibodies.

The recombinant protein obtained was then used t immunise New Zealand white rabbits, with pre-immune ser being harvested prior to immunisation. Following a boost the rabbits were sacrificed and sera collected. This ser was used in Western blots, ELISA and animal protectic models.

Using the sera obtained from the animal studies immunosorption studies were carried out.

Group B Streptococcus was grown in 20ml Todd Hewit broth (THB) for 8 hours, harvested and resuspended in Sn PBS. 50pl aliquots of this were used to coat wells in a 9 well plate (Nunc Immuno-Sorb). This was left at 4' overnight to allow for adsorbance of the bacteria onto th plate. Plates were washed twice with PBS, then blocke with 3%BSA in PBS for lhr at 37 0 C. Plates were aga washed. Serial 10 fold dilutions of the sera were made PBS and 501l of these dilutions were added to the wells i the plate, in duplicate. The plate was covered ai incubated for I hr at 37 0 C. The plate was washed, thi 501 anti-rabbit alkaline phosphatase conjugated secondal antibody at a concentration of 1:5000 was added to eat well. Following incubation at 37°C for an hour, the plal was washed again. 50Al substrate (PNPP) was added to eac well, and the reaction allowed to proceed for 30min befoz the adsorbance was read at 405 nm.

Animal protection studies were also carried out t test the effectiveness of protection'on the immunise rabbits.

GBS M732 was grown up in THB until mid-log phase :w reached approximately 5'hours. Cells were counted in counting chamber, and bacteria were diluted to give concentration of 2x10 7 bacteria per ml in pre-immune c test sera. 50pl of this was injected via tt intraperitoneal route into 0-1 day old mice. The mice wer observed for survival over 48 hours.

The following Examples illustrate the invention.

ExamlJe 1 A first clone contained a gene sequence identifie herein as SEQ ID NO. 1, with an amino acid sequenc identified as SEQ ID NO. 2, and classified as phol-13.

A comparison of the amino acid sequence of phol-13 wa performed.

Homologues to the GBS phol-13 gene product can b identified in Streptococcus pyogenes, S. pneumoniae, S salivarius, Escherichia coli, Yersinia enterocolitica Aguifex aeolicus, Hellcobacter pylori and Baemophilu Influenzae. The S. pyogenes and S. pneumoniae homologue were identified from genome sequence data and n annotations were available as to the identity of the gen or gene products. In all other cases, the above homologue 9 can be identified as ATP-dependent Clp protease proteolyt subunits. The catalytic activity of Clp proteases resul in the hydrolysis of proteins to small peptides in t presence of ATP and magnesium (Giffard, P.M. et al. 199 J. Gen. Microbiol. 139:913-920). Furthermore, the Cl component of Clp proteases has been shown to be induced part of the heat shock response (Kroh, H.E. and L.D. Simo 1990. J. Bacteriol. 172:6026-6034) and it is probable th this subunit or the complete proteolytic domain wou associated with the bacterial surface.

Immunisation studies, carried out as described abov yielded the following results.

Treatment No animals No animals surviving at time (hrs) 24 48 PBS 10 7 0 Pre-immunised 37 13 0 Immunised 38 17 9 Example 2 A second clone was selected containing a plasm designated phol-14. This plasmid contained a gene (or pa: thereof), which complemented the leaderless phoA. T2 nucleotide and deduced amino acid sequences.are shown SEQ ID NOS. 3 and 4, respectively.

A comparison of the amino acid sequence of phol-14 w.

performed.

Homologues to the GBS phol-14 gene product can identified in Streptococcus pyogenes, Enterococcus faecal.

and Streptococcus pneumoniae. These homologues we: identified from genome sequence data and no annotatiol were available as to the identity of the gene or gei products. Additionally, two possible homologues were ali identified from Shigella flexneri (SpaR) and Yersiz -pseudotuberculosls -These latter two homologues a related proteins, believed to be anchored in the bacteri membrane (Bergman, T. et al. 1994. J. Bacteriol. 176:261 2626). In S. flexneri, the product of the spaR gene h been shown to be important for invasion of epithelial cel (Sasakawa, C. et al. 1993. J. Bacteriol. 175:2334-2346 Furthermore, the product of the spaR gene is also requir for surface presentation of invasion plasmid antigens. I analogous protein in Y. pseudotuberculosls is a compone of the Yop. secretion system and is also important f virulence in this organism.

Example 3 A third clone was selected containing a plasn designated phol-5.. This plasmid contained a gene (or pa thereof), which complemented the leaderless phoA. 1 nucleotide and deduced amino acid sequences are shown SEQ ID NOS. 5 and 6.

A comparison of the amino acid sequence of phol-5 performed.

Homologues to the GBS phol-5 gene product can only identified in Streptococcus pyogenes and Staphylococc carnosus (sceA). The S. pyogenes homologue was identif from genome sequence data and no annotations were availak as to the identity of the gene or gene product Furthermore, little information is available on t function of the sceA gene product from S. carnosus. I sceA gene product shows some sequence similarity to t aggregation promoting protein from Lactobacillus gassex Based on analysis of the sceA gene product, this molecu contains a well-conserved signal sequence and is apparent secreted or associated with the bacterial cell surface.

Example 4 A further clone was selected containing a plasn designated pho3-3. This plasmid contained a gene (or pa thereof), which complemented the leaderless phoA. 1 nucleotide and deduced amino acid sequences are shown SEQ ID NOS. 7 and 8.

A comparison of the amino acid sequence of pho3-3 u performed.

Homologues to the GBS pho3-3 gene product can identified in Streptococcus mutans (rmic), (cpsM) pneumonlae and S. pyogenes. The S. pyogenes homologue w identified from genome sequence data and no annotatio were available as to the identity of the gene or ge product. In S. pneumoniae, the homologue can be identifi as dTDP-4-keto-6-deoxyglucose-3,5-epimerase. In the oth two cases, the above homologues can be identified as dTD 4-keto-L-rhamnose reductase (rmlC). In S. mutants, t gene encoding this enzyme, rmlC, is part of the rml locu The rml locus consists, of .three genes which exhib significant similarity to enzymes involved in t biosynthesis of dTDP-rhamnose, the immediate precursor the rhamnose component in the S. mutans polysacchari, capsule (Tsukioka, Y. et al. 1997. J. Bacteriol. 179:112 1134). An analogous locus has also been identified in pneumoniae (Coffey, T.J. et al. 1998. Mol. Micobiol. 17:7: 83). Almost all Streptococci characteristically posse rhamnose in their cell wall associated polysaccharid (Schleifer, K.H. and R. Kilper-Blz. 1987. Syst. App: Microbiol. 10:1-19), and it is highly probable that dTDPketo-L-rhamnose reductase would be associated with tl outer surface in Streptococci.

Example A further clone was selected containing a plasm: designated pho2-10. This plasmid contained a gene (or pai thereof), which complemented the leaderless phoA.

The nucleotide sequence is shown as SEQ ID NO.

From this, upstream and downstream coding regions wei identified, and the deduced amino acid sequences shown 9 SEQ ID NOS. 10 and 11.

A comparison of the amino acid sequences of pho2-3 was performed.

Homologues to the GBS pho2-10 gene product can I identified in Streptococcus pyogenes, Enterococcz faecalis, Debaryomyces occidentalis (hatl) and Escherlcha colil (trkD). The S. pyogenes and E. faecalls homologue were identified from genome sequence data and r annotations were available as to the identity of the ger or gene products. In the yeast D. occidentalis, the hal gene is a homologue of the trkD gene from E. col (Banuelos, M.A. et al. 1995. EMBO J. 14s3021-3027). Th trkD gene of E. coli is part of the kup potassium uptak system. The specific homolog identified here is the ku system potassium uptake protein. The kup system is constitutive potassium uptake system in EB doli. The ku system -potassium uptake protein contains a highl hydrophobic N-terminus that is predicted. to -span th membrane at least 12 times. Kup is not homologous to othe known membrane protein sequences. There is no indicatio of ATP binding, and it is proposed that the system i driven by a chemiosmotic gradient (schleyer, M. E.P Bakker, 1993. J. Bacteriol. 175:6925-6931).

Examale 6 A further clone was selected containing a plasmi designated pho2-15. This plasmid contained a gene (or par thereof), which complemented the leaderless phoA. Th nucleotide and deduced amino acid sequences-of the gene ar shown as SEQ ID NOS. 12 and 13.

A comparison of the amino acid sequence of pho2-15 wai performed.

Homologues to the GBS pho2-15 gene product can b4 identified in Streptococcus pyogenes, Streptococcus pneumoniae, Enterococcus faecalis and Escherichia col: (gatC and SgcC). The S. pyogenes, S. pneumonlae and E faecalls homologues were identified from genome sequenc data and no annotations were available as to the identit3 of the gene or gene products. In E. coli, the gatC anc sgcc gene products can be identified as being the II( component of phosphoenolypyruvate-dependent sugai 13 phosphotransferase systems (PTS), a major carbohydra -active-traniport system. I iPTS systems, the IIC compone is typically involved in binding of extracellul carbohydrates and forms a complex with the IID component constitute a membrane channel (Nobelmann, B. and J.

Lengeler. 1995. Biochim. Biophys. Acta 1262:69-72).

Example 7 A further clone was selected containing a plasm designated pho2-2. This plasmid contained a gene (or pa thereof), which complemented the leaderless phoA. T nucleotide and deduced amino acid sequences of the gene a: shown as SEQ ID NOS. 14 and 15, respectively.- A comparison of the amino acid sequence of pho2-2 w, performed.

15 Homologues to the GBS pho2-2 gene product can identified in Enterococcus faecalis, Escherichia coll (ma and afuC), Bacillus subtilis (glnO), Haemophilus influen (yebM and potA), Streptococcus pyogenes, Streptococc pneumoniae and Salmonella typhmiurlum (malK). The faecalis, S. pyogenes and S. pneumoniae homologues we: identified from genome sequence data and no annotatioi were available as to the identity of the gene or gel products. In all other cases, homologues represented AT; binding transport proteins that are part of ABC ty] transporters. Many of the components of ABC ty] transporters are membrane or cell surface associated, i these systems are involved in the transport 4 macromolecules from the extracellular environment to t intracellular compartment.

Example 8 A further clone was selected containing a plasm: designated pho3-14. This plasmid contained a gene (or pal thereof), which complemented the leaderless phoA. TI nucleotide and deduced amino acid sequences of the gene ai shown as SEQ ID NOS. 16 and 17.

A comparison of the amino acid sequence of pho3-14 wa performed and no homologues could be identified in any the public databases. One homologue to the GBS pho3gene-product can--be--identified--in Streptococcus -pyogene but this homologue was identified from genome sequence da and no annotations were available as to the identity of t gene or gene product. Using this S. pyogenes homologue search the public databases yielded no further informatio Since the pho3-14 product complemented the leaderless ph gene, it can be concluded that this protein (or pa thereof) would most probably be located extracellularly Examele 9 A further clone was selected containing: a plasm designated pho3-17. This, plasmid contained a gene. (or pa thereof), which complemented the leaderless phoA. -T nucleotide and deduced amino acid sequences of the gene a shown as SEQ ID NOS. 18 and 19.

A comparison of the amino acid sequence of pho3-17 w performed.

Homologues to the GBS Pho3-17 gene product can identified in Streptococcus mutans and Lactococcus lacti with similarity being shown to N-acetyl muramidas, Similarity is also seen with an unidentified gene, yu: from Bacillus subtilis.

N-acetylmuramidase is an autolysin that is involved.

cell division. Using this limited information along wi' the fact that pho3-17 complemented the leaderless phb gene, it can be concluded that the pho3-17 product wou: most probably be located extracellularly.

Example A further clone was selected containing a plasm: designated pho3-18. This plasmid contained a gene (or pal thereof), which complemented the leaderless phoA. TI nucleotide and deduced amino acid sequences of the gene a] shown as SEQ ID NOS. 20 and 21.

A comparison of the amino acid sequence of pho3-18 wi performed.

Homologues to the GBS pho3-18 gene product can I identified in Streptococcus pyogenes and Streptococct pneumoniae. These homologues were identified from genoa sequence data and no annotations were available as to tt identity of the gene or gene products. Using these S pyogenes and S. pneumoniae homologues to search the publi databases showed some similarity to outer surface ar membrane spanning proteins. Since the ORF3-18 produc complemented the leaderless phoA gene, it can be conclude that this protein (or part thereof) would most probably k located extracellularly.

Exalmle 11 A further clone was selected containing a plasmi designated pho3-1. This plasmid contained a gene (or par thereof), which complemented the leaderless phoA. Th nucleotide and deduced amino acid sequences of the gene ar shown as SEQ ID NOS. 22 and 23.- A comparison of the amino acid sequence of pho3-1 wa performed.

Homologues to the GBS pho3-1 gene product can t identified in Streptococcus pyogenes, Streptococcu pneumoniae, Bacillus subtills (yutD) and Enterococcu faecalis. The s. pyogenes, S. pneumonias and E. faecall homologues were identified from genome sequence data and V annotations were available as to the identity of the ger or gene products. In B. subtilis, the function of the yut gene product is unknown. It can be noted however, that tb yutD gene is located on the B. subtilis chromosome in region containing genes involved in cell wall synthesis The fact that this DNA sequence complemented the leaderles phoA gene suggests that this gene product 1 extracellularly located.

Example 12 A further clone was selected containing a plasmi designated pho3-21. This plasmid contained a gene (or par thereof), which complemented the leaderless phoA. Th nucleotide and deduced amino acid sequences of the gene ar shown as SEQ ID NOS. 24 and A comparison of the amino acid sequence of pho3-21 w performed. Homologues to the GBS pho3-21 gene product can identified in Streptococcus pyogenes, Streptococc pneumoniae, Lactobacillus fermentum (bspA) a: Lactobacillus reuteri (cnb). The S. pyogenes and pneumoniae homologues were identified from genome sequeni data and no annotations were available as to the identi" of the gene or gene products. In L. fermentum, the bs) gene product has been identified as being a basic ce: surface-located protein that has some sequence similaril to family III of the bacterial solute-binding proteil (Turner, M.S. et al. 1997. J. Bacterio1, 179:3310-3316; In:L. reuteri, the cnb gene product has been identified i a collagen binding protein that has some sequen( similarity to the solute-binding component of bacterial Al transporters (Roos, S. et al. 1996. FEMS Microbiol. Letl 144:33-38).

Examale 13 A further clone was selected containing a plasmJ designated pho3-22. This plasmid contained a gene (or pai thereof), which complemented the leaderless phoA. T1 nucleotide and deduced amino acid sequences of the gene ai shown as SEQ ID NOS. 26 and 27.

A comparison of the amino acid sequencb of pho3-22 wt performed.

Homologues to the GBS pho3-22 gene product can k identified in Enterococcus faecalis, Streptococct equisimilis (IppC), Pseudomonas fluorescens (oprI) ar Streptococcus thermophilus (orf142). The E. faecalI homolog was identified from genome sequence data and r annotations were available as to the identity of the ger or gene products. In S. equisiillis, the IppC gene produc has been identified as being a lipoprotein that i homologous to the E(P4) outer membrane protein frc Haemopbilus influensae (Gase, K. et al. 1997. Ned Microbiol. Immunol. 186:63-73). Likewise, the P 17 fluorescens opri gene encodes a major outer membral lipoprotein (Cornelis~,P.- et al. 1989. Mol. MicrobioJ 3:421-428). In S. thermophllus, the orf142 product hi been putatively identified as a cell surface expose lipoprotein that may act as a receptor for tU bacteriophages TP-J34 and Sfi21 (Neve, H. at al. 199E Virology 241:61-72). The ORF3-22 product showed goc similarity to the above homologues, particularly at th N-terminus. This is most likely the region required fc complementation of the leaderless phoA gene, and therefor serves as a leader sequence.

Examle 14 A further clone- was selected containing a plasmi designated pho3-23. This plasmid contained a gene (or par thereof), which complemented the leaderless phoA. Th nucleotide and deduced amino acid sequences of the gene are shown as SEQ ID NOS. 28 and 29.

A comparison of the amino acid sequence of pho3-23 wa performed.

Homologues to the GBS pho3-23 gene product can b identified in Streptococcus pyogenes, Streptococcu pneumonlae, Enterococcus faecalls and Streptococcus mutan (perM). The S. pyogenes, S. pneumoniae and B. faecall homologues were identified from genome sequence data and n annotations were available as to the identity of the gen or gene products. In S. mutans, the perM gene product ha been presumptively identified as a permease, but no othe: information is available as to the function of thi protein. Considering that the pho3-23 coding regio: complements the leaderless phoA gene, it can be conclude that the pho3-17 gene product would most probably b located extracellularly.

Example 1 A further clone was selected containing a plasmi designated pho3-24. This plasmid contained a gene (or parl thereof), which complemented the leaderless phoA. The nucleotide and deduced amino acid sequences of the gene are shown as SEQ ID NOS. 30 and 31.

A comparison of the amino acid sequence of pho3-24 wa performed.

Homologues to the GBS pho3-24 gene product can b identified in Streptococcus mutans (dltB), Streptococcus pneumoniae, Streptococcus pyogenes, Enterococcus faecalis, Lactobacillus casel (dltB) and Bacillus subtills (dltB).

The S. pneumoniae, S. pyogenes and B. fdecalis homologues were identified from genome sequence data and nc annotations were available as to the identity of the gene or gene products. In S. mutans, L. casei and B. subtilis, the dltB gene product has been identified as being'a basic membrane protein that is involved in the transpdrt of activated -D-alanine through the cell membrane. The dlt gene product is involved in the biosynthesis of D-alanyllipoteichoic acid (Heaton, M.P. and P.C. Neuhaus. 1992. J.

Bacteriol. 174:4707-4717). In L. casei and B. subtills, the dltB gene product is believed to contain at least 9 membrane spanning domains, indicating that the protein or portions thereof are exposed to the outside of the cell.

Example 16 A further clone was selected containing a plasmid designated pho3-29. This plasmid contained a gene (or part thereof), which complemented the leaderless phoA. The nucleotide and deduced amino acid sequences of the gene are shown as SEQ ID NOS. 32 and 33.

A comparison of the amino acid sequence of pho3-29 was performed.

Homologues to the GBS pho3-29 gene product can be identified in Borrelia burgdorferi (p23 or ospC), Bacillus brevis (owp) and Pseudomonas aeruginosa (oprI). Although these homologues are not related to each other, they all represent major outer surface proteins. In B. burgdorferi, the ospC gene product has been identified as being a 23-kDa protein that is the immunodominant antigen on the surface of this bacterium (Padula, S.J. et al. 1993. Infect. Immun.

19 61:5097-5105). The owp gene product from B. brevis is on of_tw. major cell wall proteins involved in the surfac layer lattice (Tsuboi, A. 1988. J. Bacteriol. 170:935-945) Finally, the opri gene from P. aeruginosa encodes a majo outer membrane lipoprotein precursor (Saint-Onge, A. et al 1992. J. Gen. Microbiol. 138:733-741).

Example 17 A further clone was selected containing a plasmil designated pho3-50. This plasmid contained a gene (or par thereof), which complemented the leaderless phoA. Thl nucleotide and deduced amino acid sequences of the gene ar, shown as SEQ ID NOS. 34 and A comparison of the amino acid sequence of pho3-50 wal performed..

Homologues to the GBS pho3-50 gene product can b identified in a variety of Streptococci (penA, pbp2B pbpB2), Borrelia burgdorferi (pbp2), Enterococcus faecalil (pbpC), Staphylococcus aureus (pbpA), Mycobacterlum leprai (pbpB) and Helicobacter pylori (pbp2). In all cases, th above homologues can be identified as penicillin bindini proteins (PBPs).. Genes encoding penicillin bindin< proteins are often located in a cluster of genes associatei with cell wall synthesis (Pucci, M.J. et al. 1997. J Bacteriol. 179:5632-5635). Furthermore, PBPs are typicall: integrated into the cell wall of a bacterium with some o0 all of the protein being exposed on the outer bacteria: surface.

Claims (14)

1. A peptide when used in therapy encoded by the gene identified herein as pho3-21, obtainable from Group B Streptococcus, or a homologous peptide having at least sequence similarity thereto or an immunogenic portion thereof, wherein the homologous peptide or the immunogenic portion elicits a protective immune response against Group B Streptococcus.

2. A peptide according to claim 1, comprising the amino acid sequence identified herein as SEQ ID NO.

3. A polynucleotide encoding a peptide according to claim 1 or claim 2, when used in therapy.

4. A host transformed to express a peptide according to claim 1 or claim 2. A vaccine comprising a peptide according to claim 1 or claim 2, or the means for its expression.

6. Use of a peptide according to claims 1 or 2, a polynucleotide according to claim 3 or a host according to claim 4, for screening potential drugs or for the detection of virulence.

7. Use of a peptide according to claims 1 or 2, a polynucleotide according to claim 3 or a host according to claim 4, for the manufacture of a medicament for use in the treatment or prevention of a condition associated with bacterial infection.

8. Use according to claim 7, wherein the infection is a Group B streptococcal infection.

9. Use according to claim 7 or claim 8, wherein the infection is a focal infection. IO 21 Use according to claim 7 or claim 8, wherein the infection is a urinary tract infection.

11. An isolated antibody raised against a peptide according to claim 1 or claim 2.

12. A peptide substantially as herein described with reference to Example 12.

13. The peptide according to claim 1, wherein the sequence similarity is at least SEQUENCE LISTING <110> Microscience Limited <120> GENES AND PROTEINS, AND THEIR USE <130> REP05973WO <140> <141> <160> <170> PatentIn Ver. 2.1 210>, 1 <211> 587 <212> DNA <213> "group B streptococcus <220> <221> CDs <222> (582) <400> 1 atg ate Met 11e. 1 cca gta gta atc gaa caa aca agt cgt ggt gaa cgt tct tat Pro Val Val Ile Glu Gin Thr Ser Arg Gly Glu Arg Ser Tyr gat att tac tea Asp Ile Tyr Ser cgt ctt tta aaa Arg Leu Leu Lys cgt att att atg Arg Ile Ile Met ttg aca ggc Leu Thr Gly caa gtt gag gat aat atg goc aat agt ate att gea cag tta ttg ttt Gin Val Glu Asp Asn Met Ala Asn Ser Ile Ile Ala Gln Leu Leu Phe ctc gat gca caa gat aat aca Leu Asp Ala Gln Asp An Thr aag gat att tac ctt tat gtc aat aca Lys Asp Ile Tyr Leu Tyr Val Ann Thr cca ggt ggt tca gta tog get gga ctt get att gtg gac acc Pro Gly Gly Ser Val Ser Ala Gly Leu Ala Ile Val Asp Thr 70 75 ttc att aaa tog gac gta cag acg att gtt atg ggg atg get Phe Ile Lys Ser Asp Val Gin Thr Ile Val Met Gly Met Ala atg aac Met Asn get tog Ala Ser atg gga acc Met Gly Thr tta cog sat Leu Pro Aa 115 gga ggt aca. Gly Gly Thr 130 ama acg cgt Lys Thr Arg 145 tot att gas Ser 1e Glu get cam gaa Ala Gin Gl sat gaa tax Asn Glu <210> 2 <211> 194 <212> PRT <213> group <400> 2 Net Ile Pro 1 Asp Ile Tyr Gia Val Gln Leu Asp Ala Pro Gly Sly a I 1 A ci 9; c t: as ILr a TI II att get tca Ile Ala ser gas tat atg Glu Tyr Met caa tot gat Gin Ser Asp 135 act tta gas Thx Leu Glu 150 gtc cat gat Val His Asp 165 ott gat tat Leu Asp Tyr ggt get aaa gga aaa Gly Ala Lys Gly Lys 105 cac caa cca atg gge His Gin Pro Met Gly 125 get ate get get gag Ala Ile Ala Ala Glu 140 ate tta get gat aat Ile Leu Ala Asp.Asn 155 gca gag cgt gat cgt Ala Glu Arg Asp Mrg 170 ttt att gat gaa ate Phe Ile Asp Glu Ile 185 A AL fw".7IVI.Fa0 I cgt ttt atg 336 Arg Phe Net 110 gga aca gge 384 Gly Thr Gly cat ott tta 432 His Leu Len tct ggt caa 480 Ser Gly Gin 160 tgg. atg agt 528 Trp Met Set 175 atg get aat 576 Met Ala Asn 190 B streptococcus Val Val Ile Glu 63 5 Ser Arg Len Len Ll Asp Asn Met Ala As 4 Gln Asp Asn Thr Ly 55 Ser Val Ser Ala G1 Thr Ser Asp Arg Set Ile Asp Ile Leu Ala Arg Gly Ile Ile Ile Ala Tyr Lou Ile Val WO 00/37646 Phe Ile Lys Hot Gly Thr Leu Pro Asn 115 Gly Gly Thr 130 Lys Thr Arg 145 Sex Ile Glu Ala Gln Glu As Glu Ser Ile 100 Ala Gin His Lys rhr Lso Asp 85 Ile Glu Gin Thr Val 165 Leu val Gln Thr Ala Ser Ser Tyr Met Ile 120 etr Asp Met 135 Leu Glu Lys 150 His Asp Asp Asp Tyr Gly Ile Gly 105 His Ala Ile Ala Phe 185 75 Val Met Gly Met Ala Ala Ala Lys Gly Lys Arg Phe 110 Gin Pro Met Gly Gly Thr 125 Ile Ala Ala Glu His Leu 140 Leu Ala Asp Asn Sec Gly 155 Glu Arg Asp Arg Trp Net' 170 175 1e Asp Glu Ile Met Ala 190 PCT/GB99/04377 ser Met Gly Leu Gln 160 Ser An <210> 3 <211> 218 <212> DNA <213> group B streptococcus <220> <221> CDS <222> <400> 3 ate aga gca tat tot ggt cct ctt tog gtt ttc ctg cca cgt ttt aaa Ile Arg Ala Tyr Ser Gly Pro Leu Ser Val Phe Leu Pro Arg Phe Lys 1 5 10 get tgt gat ate ata gtc aat gtg agg agg act'atc atg tta ttt aag Ala Cys Asp Ile Ile al Asn Val Arg Arg Thr Ile Met Leu Phe Lys 25 gaa aaa att cot gga cta ata tta tgc ttt att att get ata cca tot Glu Lys Ile Pro Gly Leu Ile Leu Cys Phe Ile Ile Ala Ile Pro Set 3 yIS UUIJJIrU tgg ttg ctt Trp Lou Leu rL Aitwyna I I ggg ctt tat ctc cct Gly. Lou Tyr Lou. Pro 55 tta atm ggm gca cca gto ttt got Leu Ile Gly--Ala Pro--Val- Phe Ala ito ttg att Ile Lou Ile <210> 4 <211> 72 <212> PRT <213> group gga atm att gtt ggm to Gly Ile Ie Val Gly B utroptococcus <400> 4 fle Arg 1 Ala Tyr Sor Sly Pro Lou Sot 5 rho Lewi% -Pro, Arg Phe Lys Ala Cys Asp Ile Ile Val Asn Val Arg Arg Thi le oet Leu Phe Lys Ile Pro 8cr Glu Lys Ile Pro Gly Lou Ile Lou Cyu Phe Ile Ile TXp Lou Ile Leu <210> Lou Sly Lou Tyr Lou Pro Lou fIl ely Pro Val Phe Aia Ile Sly lie Ilo Val Gly <211> 705 <212> DNA <213> group B streptococcus <220> <221> CDS <222> <400> atg aat ear aga aga aaa tta tea Met Asn LyS Arg -Arg Lys Lou 8er mum ttg mat gtm ama ar cam cat LYS Lou Asn Val Lys Lys Gin His is C CIYYI~ V~I tta get tat ggm get ate act tta gta gee ett ttt tea Leu Ala Tyr Gly Ala Ile Thr Leu Va1 Ala Len Phe Ser 25 tgt att ttg Cys Ile Len pet pta acg ptc ate ttt ama apt tea cam Ala Va1 Thr Val Iie Phe Lys Sr 5cr Gin 40 ptt act act gaa tet ttg Val Thr Thr Gin Set Len tea aa gca Ser Lys Ala pat aaa gtt Asp Lys Val gta gee aaa aaa Val Ala Lye Lys ama atp act aag Lys Met Thr Lys peg aca tet mai tea Ala Thr ser Lye Sex gta gam gat gta Val Glu Asp Val cap get cea aaa Gin Ala Pro Lys tot cap gem tet mat gam gee cca aaa 8cr Gin Aa Ser Ann Glu AlaPro Lye apt tet caa tt Sex Ser Gln Set moa ga 288; Thr, Glu get mat tot Ala Asn Sr cam gca gtt Gin AMa Val 115 cam cam gtt act Gin Gin Val Thr apt gaa gag acg Ser Glu Glu Thr get gta. ga Ala Val Glu 110 cac mac aac His Ann Asn *336 384 gta aca ga atm Val Thr Glu Iie ctg cta coi gte Len Len Pro Val agg Ar; 125 ctt tat Len Tyr 130 get gtt met gag Ala Val Thr Glu ect tn aac ct get cam cca cca ge Pro Tyr Asn Pro Ala Gin Pro Pro Asp 140 ptg gee mpg tat Val Ala Arg Tyr gag Gin 150 caa tg ama tac Gin ?rp Lys Tyr cap gcg gte Gin Aim Val ppm tct Giy 8cr 160 tct act ser Thr 175 get get gem gem cam atg get got gca acm gga gtc cet eag ely Val Pro Gin Ala Ala Ala Ala Gin 165 Net Ala Ala Aim. Thr 170 tgg gaa cat att att gee ept gam tea mat gpt mat Trp Gin, His Ile Ala Arg Glu Asn Giy Asn cct mat gtt pet Pro Asn Vai Ala 190 mat pee tea pga get tea gga ett tto caa ae atg eca gpt tpg gpt Asn Aim Ser Gly Ala Ser Gly Leu Phe Gin Thr Met Pro ely Trp Gly 195 200 205 tca aca got Ser Thr Ala 210 cgt gct ceaa Arg Ala Gin 225 <210> 6 <211> 234 <212> PRT <213> group aca gtt cag gat ceaa gta att cag cta tta aag ctt att 672 Thr Val Gin Asp Gin Val Ile Gin Leu Leu Lys Leu Ile 215 220 ggg tta toa got ggg tac cag tga 705 Gly Leu Ser Ala Gly Tyr Gin 230 235 B streptococcus <400> 6 Met Asan .1 Leu Ala Ala Val Ser Lys Ala Thr Ser Gin Ala Asn Gin Ala Leu Tyr 130 Gln Val 145 Lys Arg Arg 5 Tyr Gly Ala Thr Val Ile Ala Asp Lys Set Lys Ser Ala Ser Asn Ser Gin Gin 100 Val Val Thr 115 Ala Val Thr Ala Arg Tyr Lys Leu Ser Ile Thr Leu Phe Lys Set 40 Val Arg Val Lys Val Glu 70 Glu Ala Pro Gin Val Thr Glu Ile Pro 120 Glu Thr Pro 135 Glu Gln Trp 150 Lou Asn 10- Ala Lau Gin Val Lys Lys Val Lys 75 Set Ser 90 Ser Glu Lou Pro Asa Pro Tyr Cys 155 LyS Lys Gin Seercys Zle: Thr Gli Ser Lys Met Thr Ala Pro Lys Gin Ser Thr Thr Ala Val 110 Arg His Asn 125 Gin Pro Pro Ala Val Gly His, Leu ,Lys Pro Glu Glu Asn Asp Ser 160 Ala Ala Ala Ala Met Ala Ala Ala Thr 170 Gly Val Pro Gin ser Thr 175 Trp Glu His 10 180 Asn--Ala Sex Sly 195 I. Ala Arg Giu Sbr Ann Gly 195 Asn Pro Asn Val Ala 190 Ala Ser Gly Lau -Pho 200 Gin Thr Met Pro Gly Trp Gly 205 Leu Lys Lau Ile sex Thr Ala Thr Val Gin Asp 210 215 Arg Mla Gn Gly Lau Ser Mla 225 230 Gin Val Ile Gln Sly Tyr ln <210> 7 <211> .594 <212> D1A <213> group B streptococcus <220> <221> CDS <222> <400> 7 atg act gam cc& ttt Met Thr Gl Pro Phe 1 5 ttt gat aaa gaa Phe Asp Lys lu act tgt ago cca Thr Cys Atg Pro ctt gas Ile Glu gao att cat gas Ala Ile Pro Glu ogt ggs tgg ttc Arg Cly Txp Ph. ttg ttg gas ttc Lou Lou Glu Ph. stt ace gtt cgt Ile Thr Val Arg ggc gac sac ely Asp Ann aaa gag sac Lys Glu Ann cas ass gas aaa atg ate ccg ott Gin Lys lu Lys Met Ile Pro Lau ggt ttc ely Ph. cos gsa ago Pro GIM Ser ttc ttt Pho Ph. gag gca gsc aac Glu Ala Asp Lys cas ast sat att Gin Ann Ann Ile 192 tog ttt acs asa aaa cat act ttg cge ggt ctc Sex Ph. Thr Lye Lys Ann Thr Lou Arg Sly Leu 70 cat gca gag cot His Ala Glu Pro gat aaa tat gtt Asp Lys Tyr Val tcg atc got gat gsa gga cgt gtg Sex Xle Ala Asp Glu Sly Arg Val atc ggt act tgg Ile Gly Thr ?rp gtt Val ate Ile ttc Ph* tat Tyr 145 gat Asp' tea Ser ag Lye ctc Len Ala 115 gtt val 9ca Ala mat Ash Ala gaa 195 cgt Arg 100 tea Set ctt Leu Ctt Leu cta Leu gat Asp 160 gat Asp gas ggt Siu Sly aaa ggt Lys Sly tea got 8er Asp gas etc Glu Leu 150 ggc att Sly Ile 165 aag mat Lys Ran ttg tee Lou egt ttt Ser Phe 105 ttt qtt The Val 120 gem get Ala Ala caa aea Pro Lye tgg gaa Trp GIU aca ctt Pro Lou 185 ggt sac gtt tec caa Sly axn Val Tyr GIn 110 oca cgo gge gtt got Pro Arg Gly Val Ala 125 tat act tat ctc gtt Tyr Thr Tyr Lou Val 140 tat got ttc gtt aee Tyr Ala Phe Val Ran

155. mat etw gas gaa gca Asn Xa Glu Gin Ala 170 etc aa gat gte aaa Lou Lys Asp Val Lys 190 acg Thr aat Asn mac Anna tat Tyr gaa Glu 175 act. Pro att 336 Ile ggt 384 Gly get 432 Ap gca 480 Ala 160' gtc. 529 Val ttg 576 Leu <210> 8 <211> 197 <212> PRT <213> group <400> 8 Met Thr GlU 1 Ala Ile Pro Arg Sly Trp Sly The Pro 3cr Pbs Thr a streptococcus Pro Phe Phe Asp 5 Sin Lou Leu Slu Phe Lys Glu Asn GlU Set Phe Phe Lye Lye Ran Thr Lys Sin Phe Asp Phe Gin Ginu Ala Leu Arg 8 Ile Asp Pro Asn Pro Asp Val Ile Phe Tyr 145 Asp. Ser Lys Tyr Leu Ala 115 Val Ala ,Asn Ala Glu 195 Val Arg 100 Ser Leu Leu Leu Asp 180 Asp 70 ser Ile Glu Gly Lys Sly Ser Asp Glu Leu 150 Gly Ile 165 Lys Aan Leu 75 Gli Gly Arg Val 90 Phe Gly Asn Val 105 Val Pro Arg Gly Ala Tyr Thr Tyr 140 Lys Tyr Ala Phe 155 Glu Asn Xaa Glu 170 Leu Leu Lys Asp 185 Gly Thr Trp Gin Thr lHe 110 Ala Asn Gly Val Asn Asp Asn Tyr Ala 160 Ala Glu Val 175 Lys Pro Leu 190 <210> 9 <211> 1217 <212> DNA <213> group B streptococcus <220> <221> CDS <222> <220> <221> CDS <222> (679)..(945) <400> 9 tat tat tta ate gga ggg ttg gca gaa atg caa cat gtc aat cat tct Tyr Tyr Leu Ile Gly Gly Leu Ala Glu Met Gin His Val Asn His Ser 1 5 10 tct ttt gat aaa gca tea aaa gca gga ttt att att get tta ggc att Sex Phe Asp Lys Ala Ser Lys Ala Gly Phe Ile Ile Ala Leu Gly Ile 9 gtt tat gg&, gut att ggt mon Vul Tyr sly Asp Il. Gly Tb: ago oca atc tat ag utg aa ta ttg So: Pro Lou Tyr Tb: Mot Gin So: Lou gtt qua usc can ggt ggt att tot agt gtc aca gem tog ttt into tta Vul Giu Ann Gin Gly Sly ie Sex So: Val Tb: Giu So: Pb. Ile Lou tot at& tot ttu ato atm tgg Be: I1e Se: Lou Ile Ii. Trp uce ttg non Tmx Lou Th: ott utt uca act Lou Ile Tb: Tb: aug tat gtg ott gtu got ttu aag gog gut nut cac oao gnu Lys Tyr Val Lou Val Alu Lou Lys Ala Asp Asn His His (flu 09t ggt Gly 61y- utt ttt tat Ile Phe Ser gtt cog gat Val Pro Ala 115 ttm tat uco ctt Lou Tyr Thr Lou 100 gtt att gga ggt Val Ile Sly Gly qtt agu Val flg .105 uuu utg uce act Lys Not Tb: Pro tgg ttu utt Trp Lou Ile 110 gga got ttg Sly Ala Lou ace ttg ttg tca Tb: Lou Lou So: mog coo Tb: Pro 130 got gtu c gtu Ala Val Tb: Val ag cog tta egg Sin Pro Lou Arg mum gta gtt Cot Lys Val Val Pro ttg cag cat att Lou Gin His Ile aga ato age gte Arg Ile Azrg Val tat ttt gog inca Tyr Pbs Aim Thr tta ttt act gtt Lou Pb. Tb: Val act Tb: 165 ttt goc uto cam Phe Ala Ile Gin ttg gum ogg gtg Lou Slu Azrg Val tta ttg Lou Lou 175 guu ttu ttg Giu Lou Lou 900 Ala 180 att atg tta tat Ile Met Lou Tyr ggt Sly 185 ttg cat ttt ggt Lou Pro rho Sly ncggtctoot. tutagttttg coatacaga agttttoaag cattataca tactaoggtt 630 tgesettgtt atttagtcca gegaetoata aaggtetttt tettttag gut ca ttt 687 Asp Lou Pb. too tgg oga aa acg gga ga gina gon ca tao tot gao tta ggt cat 735 Trp Arg Gin Thr Gly Ala 195 200 gttggg-c-gt. gga aat a ta cat Val Gly Arg Gly Ann Ie His 210 215 att ata ott tot tat tgt ggg Ile Ile Lou Ber Tyr Cys Gly 230 ao gca gga mat gina ttg mat Ann Ala Gly Ann Giu Lou An 245 ttt so& atg oat gtc gtt att Phe Thr Met His Val Val Ile 260 tca cag gca ctg att tct gg* Set Gin Ala Lou Ile Set 275 ogactaaa tattccaca atttc' otaoatacct ggtattaatt ggtto taagacttoa gcgoaoatgg aagca' gaeaactatt tt&ctgtctt tctttl tt. <210> <211> 190 <212> PUT <213> group R streptococcus <400> Tyr Tyr Lou Ile Gly Gly Lou 1 5 Set Phe Amp Lys Ala So: Lys Vali Tyr Gly Asp Iie Gly iTh Giu Ala Lau Tyr Set Asp Lou Sly His 205 gtt toa tgg cog tto gtt aag gtt gcc Val 5cr Tip Pro Phe Vai Lys Val Ala 220 225 cam ggg ga tgg att tta gat aat sag Gin (fly Ala Trp Ile Lou Ala Ann Lys 235 240 ccc ttt ttt got sgt att coct tog cain Pro Pbs Pbs Ala Se Ile Pro Bor Gin 250 255 tta gat act ttg goa got Atca ito got' Lau Ala Thr Lou Ala.,Ala Ile Ile Ala 265 270 toaattt accttaagtt'ctgagctatg' gttca acttatcotg ttgacastat tgggtcainac ttatt tgocattaoa acotctattg gtttgotttt points tggattagog atamaatta cgatgotaat ttaat toasaaagga qtaaagagag gttttagcts 783 831 87 9 9227 975 1035 1095 1155 1215 1217 Ala (flu Met Gin His Val Ann His 8cr 10 1s Ala Sly Pb Ile Ile Ala Lou Sly Ile 25 Sex Pro Lou Tyr Th: Met Gin Ser Lou 40 11 Val Glu Asn Gin Gly Gly Ile Sex Ser Val Thr Glu Ser Phe lHe Leu 55 Gly Ser Ile Ser Leu lle Ile Trp Thr Leu Thr Leu Ile Thr Thr Ile 70 75 Lys Tyr Val Leu Val Ala Leu Lys Ala Asp Asnn His His Glu Gly Gly 90 Ile Phe Ser Leu Tyr Thr Leu Val Arg Lys Met Thr Pro Trp Leu Ile 100 105 110 Val Pro Ala Val Ile Gly Gly Ala Thr Leu Leu Ser Asp Gly Ala Leu 115 120 125 Thr Pro Ala Val Thr Val Leu Gin Pro Leu Arg Ile Lys Val Val Pro 130 135 140 Ser Leu Gin His Ile Ser Arg Ile Arg Val Cys Tyr Phe Ala Thr Leu 145 150 155 160 Leu Phe Thr Val Thr Phe Ala Ile Gin Gly Leu Glu Arg Val Leu Leu 165 170 175 Glu Leu Leu Ala Ile Met Leu Tyr Gly Leu Pro Phe Gly Leu 180 185 190 <210> 11 <211> 89 <212> PRT <213> group B streptococcus <400> 11 Asp Leu Phe Ser T rp Arg Gin Thr Gly Ala Glu Ala Leu Tyr Ser Asp 1 5 10 Leu Gly His Val Gly Arg Gly Asn Ile His Val Ser Trp Pro Phe Val 25 Lys Val Ala Ile Ile Leu Ser Tyr Cys Gly Gin Gly Ala Trp Ile Leu 40 Ala Asn Lys Asn Ala Gly Asn Glu Leu Asn Pro Phe Phe Ala Ser Ile 55 Pro Ser Gin Phe Thr Met His Val Val Ile Leu Ala Thr Leu Ala Ala 12 Ile Ile Ala Bcr Gin Ala Lou Ile 3cr <210> <211> <212> <213> <220> <221> <222> 12 378 DNA group B streptococcus C.3 (1)..(3718) <400> 12 ntg cag gta met Gin Val ttt tta sat att gtc mat nsa tto *ttt gat, cca Ph. Lou Asn Ile Val An Lye Phe Phe Asp Pro gtt att 48 yal Ile cat atg ggt tog gg gtt gtg stg His Met rly Ser Sly Val Val Met att gto atg aeA Ile Val Met Thr ggt tta -gcC Sly Lou Ala ggt att sag Gly Ile Lye atg ata ttt Met lie Phe ggn gtg sag ttt Sly Val Lye Phe tot ana gca 5cr Lye Ala 40 ott gas ggt Lou Glu Sly 144 tta got Lau Ala att gct ctt acg le Ala Lou Thr ggt att ggt got Sly Ile Sly Ala 55 ott ca got ttt Lou Gln Ala Ph. att att ggt stt tta act Ile TIe Sly Ile Lou Thr ggt got ttt too gas sly Ala Phe Scr Glu aaa &at &CA gga Lye Ann Thr Gly sat ota ago att Ann Lou er Ile gac gtt ggt tgg Asp Val Gly Trp con tta gon act Pro Lou Ala ThE att aoa Ile Thr tgg gga ton Trp Sly Set sat att gtt Arn le Val 115 tat acg ctt Tyr Thr Leu to tto Tyr Pho 105 ttn tta ato atg Lou Lou Ile Met ott stt gto Lou Ile Val 110 atg att gtt atg aaa ama an cgg ata Met Ile Val Met Lye Lye Lys Arg Ile 120 oct tag 378 Pro 125 <210> 13 <211> 125 <212> PRT <213> group B streptococcus <400> 13 Met Gin Val Phe Leu Asn Ile 1 5 His Met Gly Ser Gly Val Val Met Ile Phe Gly Val Lys Phe Leu Ala Ile Ala Leu. Thr Gly 55 Gly Ala Phe Ser Glu Set Leu 70 Asn Leu Ser Ile Ile Asp Val Trp Sly Ser Pro Tyr Thr Leu 100 As Ile Val Met Ile Val Met 115 Val Asn Met Leu 25 Ser Lys 40 Ile Gly Gin Ala Gly Trp Tyr Phe 105 Lys Lys 120 Lys P PhPhe Asp Ile Val Met Thr Ala Leu Glu Gly Ala Ile Ie Gly Phe Val Lys Asnn 75 Ala Pro Leu Ala Leu Lou Ile et Lys Arg Ile Pro 125 Pro Sly Gly Ile Thr Thr Leu 110 Ile Ala LYs Thr Hle Thr Val <210> 14 <211> 705 <212> DNA <213> group B streptococcus <220> <221> CDS <222> (118)..(705) <400> 14 ggatcgggcg caagcttaac gattcttttt aaaatcatta aattttaaaa caaatttcag acatattgcc aaagttttga tattattact ataatatagt ttgtagagga gaataat 117 14 mtp gpo can qua oct ate ate gnu tat cia sat ate Met Sly Gin Gin Pro Ile Ile Gin Tyr Gin Asn Ile nut ann gtgj tat Ann Lys Val Tyr pp9ga pin t. sly Gin An gat ttc gtt Asp Ph. Vai utg opt ntq Met Arg Met s0 gtt gap gtt gun gut att ac ott itt Val Ala Vai Gin Asp Ile Ann Lou Lys Ile tao cot ggt Tyr Pro Gly tgt tte ate pgt Cys Ph. Ile Sly apt 9ga tca ppt &nn non acou ttn 3cr Sly Soc Sly Lys Thr Thr Lou ptt nac cat ntg ttn ann cc& aoa sant ggt act eta tta Vai Ann His Hot Lou Lys Pro Thr Azn Sly Thr Leu Leu ttt Phe nag gqi put Lys Giy.Lye Asp tot act att ane Sec Thr Zle An att gai ttu Ile Gin Lou api cgc Arg Arp so atg &cc Met Thr spa att 9gn Arg Ile Sly att tac qa Ile Tyr Gin tat gtt Tyr Val ate cna ae att Ile Gin Asn Ile tta atg cot out Lau Met Pro His atu ptt ott gta XIe Val Leu Val aaa tta ttp aaa tpp tea pin Lys Leu Leu Lys Tzp Ser Gin 110 gun got nun api got aun gen app pan ott att na Gin Ala Lys Arg Aln Lys Ala Arg Gin Lou Ile Lys gtt pan tta Vai Gin Lou ccc gnu Pro Gin 130 can can Gin Gin 145 gnu tat ttg pat Giu Tyr Leu Asp tio cot apt pg Tyr Pro Sex Gin tot gpc ggt cap Ser Sly Gly Gin opt ate ggt Arg Ile Gly gtc Val 150 ntt cgo got Ile Arg Ala ott pci Lou Ala 155 gci gac cii pat Ala Asp Gin Asp itt tta atp pat Ile Lou Mot Asp cct ttt gn get Pro PhiG Gly Ala pat cot itt aot Asp Pro Ile Thir upa gnu Arg Gin 175 gqt utt eta gac ttt apt can pto tot tea 9g aga nit Sly Ile Gin Asp Pb. 3cr Gin Val 5cr 180 185 Sex Sly Arg An gqp ga aa Sly Sly Lys 190 cta tea tct tag Leu Ser Ser 195 <210> <211> 195 <212> PRT <213> group B.streptococcus <400> Met Gly Gin Glu Pro Ile Ile 1 5 Glu Tyr Gln Asa Ile Asn Lys Val Tyr Gly Asp .net Phe Arg Ile Glu Pro Gin 145 Ile Glu Asn Phe. Val Arg. Met Lys Gly Ile Gly Tyr Glu Ala Lys 115 Glu Glu 130 Gln Arg Leu Met Val Cys Val Lys Tyr Asn 100 Arg. Tyr lie Asp Ala Phe Asn Asp Val Ile Ala Leu Gly Val lie His Ile 70 Ile Val Lys Asp Val Glu Asp Gly Thr 40 Met Leu 55 Ser Thr Gln Asn Leu Val Ala Arg 120 Arg Tyr 135 Ile Arg Ile 25 Ser Lys Ile 110 Pro 105 Glu Pro Ala 10 Asn Gly Pro Asa Gly 90 Lys Leu Ser Leu Leu Leu Ser Thr Pro 75 Leou Leu Ile GIu Ala 155 Asp Lys Gly Arsn Ile Net Leu Lys Leu 140 Ala Pro Ile Lys Gly Glu Pro Lys Leu 125 Ser Asp Ile Tyr Pro Gly Thr-Thr Leu Thr Leu Leu Leu Arg Arg His Met Thr Trp Ser Glu 110 Val Glu Leu Gly Gly Gln Gin Asp Iie 160 Thr Arg Glu 175 Gly Gly Lys 190 150 Glu Pro Phe Gly Ala 165 170 Gin Asp 180 Ser Phe Ser G1n Val Ser 185 Set Gly Arg Asn <210> <211> <212> <213> <220> <221> <222> 26 361 group B streptococcus CDS (366) <400> 16 ate cot tat agt gat gtt ttt Ilo Pro Tyr 5cr Asp Val Pho get aca gga gga ttt tta tee tat gta Ala Tb: Gly Gly Phe Lou Tyr .Tyr Val cg att. got eta agt Tb: Ile Ala Len 5cr ta ctt tta Tyr Lou Lou ggg tat agt etc tggf tte ttt att Gly So: So: Iet Try Lou Ph. I1e 25 tet gga att tat ttt tat aae tte 5cr ely Ile Tyr Pbs Tyr Lys Lou gte cag ttt att Val Gin Phe Ilo get tao tat Ala Tyr Tyr gtt tat Val Tyr tat gtg gaa ae Tyr Val Ala Gin gas ett gta tog Glu Iie Val So: att ttc tat ate atg Ile Phe Tyr Ile Mot ate, ag tgg qca tta Ile Gin Try Ale Lou att gte tta gge Ile Val Lou ely ate ggcaetg ag ttg Ile Sly Met Tb: Lou ggt ggt etq tee cca Sly Sly Met Tyr Pro 00 eta tgg ttt tta att Lou Trp Phe Lou Ile 192 240 288 cat ttt etg ate Pro Pbs Net Lou att tag Ile So: s0 ac ttt tgt Lys rho Cys get eat etc gtt Asp Asn Ile Vel gsa ga ttt eta Glu Ala Phe Ile ttt tat ggt Pho Tyr Sly 110 ett tta ga gca ttc tee ota Ilo Lou Ala Ala Pb. So: Lou 115 ttt ate gat c Flt I.e Asp 120 <210> 17 <211> 122 <212> PRT <213> group <400> 17 Ile Pro Tyr 1 Thr Ile Ala Val Gin Phe Val Tyr Tyr Ile Phe Tyr 11e Gin Trp Lys Phe Cys Ile Leu Ala 115 B streptococcus Ser Asp Val Phe Ala 5 Leu Ser Tyr Leu Leu lle Ala Tyr Tyr Val Val Ala Gin Ser Glu Ile Met Asn Ile Val 70 Ala Leu Pro Phe Met Val Asp Asn Ile Val 100 Ala Phe Ser Leu Phe 120 Thr Gly Gly 10 Gly Ser Set 25 Ser Gly Ile Ile Val Ser Leu Gly Phe 75 Leu Ile Sert 90 Asp Glu Ala 105 Ile Asp Tyr Tyr Leu Phe Tyr Lys Met Thr Met Tyr Phe Leu Phe Tyr 110 Val lle Leu Leu Pro Ile Gly <210> 18 <211> 570 <212> DNA <213> group B streptococcus <220> <221> CDS <222> <400> 18 atg agg aaa cgt ttt tec ttg cta aat ttt att gtt gtt act ttt att Met Arg Lys Arg Phe Ser Leu Leu Asn Phe ie Val Val Thr Phe Ile 1 5 10 ttc ttt ttc ttt att ctt ttt ccg ctt tta aac cat aag gga aaa gta Phe Phe Phe Phe Ile Leu Phe Pro Leu Leu Asn His Lys Gly Lys Val 25 18 gat got sat tot agg cag agt gtt acc tac acc aaa gas gas ttt ate Asp Ala Asn Ser Arg Gin ser val Thr Tyr Thr Lys Glu Glu Phe Ile ca aaa Gin Lys cgt ct Arg Pro att gtg cca get Ile Val Pro Asp tea ttt att att ser Phe Ile Ile 70 gcg caa gat cta gga aag tog tac ggt att Ala Gin Asp Leu Gly Ly Ser Tyr Gly Ile 55 gca cag gcg got ttg gat tot get tte gga Ala Gin Ala Ale Leu Asp Ser Asp Phe Gly 75 gag aaa tat age Glu Lys Tyr Ser agt etc ata Ser Ile Ile atc tgt Ile Cys tgg ttg ctt gca Trp Leu Leu Ala gaa cca Glu Pro' 288

336. gg aecg ccc Gly Thi Pro gas ag caa Glu Lys Gln 115 tac ctt get Tyr Leu Ala 130 att acc tta sat Ile Thr Leu Asn gat agt agt aca gga aea aaa cag Asp ser ser Thr Gly Lys Lys Gin 105 110 ttt act cat tat Phe Thr His Tyr tot tgg aag tat Ser Trp Lys Tyr tea atg get get Ser Met Asp Asp 125 cat ate aaa His Ile Lye gga gcg aca ggc aaa aa get tea tat Gly Ale Thr Gly LyS Lye Asp Ser Tyr 140 eat cca aae act tta gtg caa aaa tta ate atg gtg Ile Met Val tot gtt aaa Set Val Lys 150 Asn Pro Lye Leu Val Gin Lys Leu 160 caa gat agt ggt Gin Asp Ser Gly gat aat gac aaa Asp Asn Asp Lye tac gt aa Tyr Ala Lys aaa atg acg Lys Met Thr 175 gaa atc att Glu Ile Ile ttg tat gat tta Leu Tyr Asp Leu aga tat gat aag tga Arg Tyr Asp Lys 190 <210> 19 <211> 189 <212> PRT <213> group B streptococcus <400> 19 Met Arg Lys Arg Phe Ser Leu Leu Asn Phe Ile Val Val Thr Phe Ile 1 5 10 Phe Phe Phe Phe Ile Leu Phe Pro Leu Leu Asn His Lys Gly Lys Val 25 Asp Ala Asn Ser Arg Gin Ser Val Thr Tyr Thr Lys Glu Glu Phe 11e 40 Gin Lys Ile Val Pro Asp Ala Gin Asp Lou Gly Lys ser Tyr Gly Ile 55 Arg Pro ser Phe Ile Ile Ala Gin Ala Ala Leu Asp Ser Asp Phe Gly 70 75 Glu Lys Tyr Ser Tyr Ser Ile Ile IIe Cys Trp Leu Leu Ala Glu Pro 90 Gly Thr Pro Ser Ile Thr Leu Asn Asp Ser ser Thr Gly Lys Lys..Gla 100 105 110 Glu Lys Gln Phe Thr His Tyr Lys ser Trp Ly Tyr ser Met Asp Asp 115 120 125 Tyr Leu Ala His Ile Lys Ser Gly Ala Thr Gly Lys Lys Asp Ser Tyr 130 135 140 Thr Ile Met Val Ser Val Lys Asn Pro Lys Thr Leu Val Gin Lys Leu 145 150 155 160 Gin Asp ser Gly Phe Asp Asn Asp Lys Lys Tyr Ala Lys Lys Met Thr 165 170 175 Glu Ile Ile Asp Leu Tyr Asp Leu Thr Arg Tyr Asp Lys 180 185 <210> <211> 978 <212> DNA <213> group B streptococcus <220> <221> CDS <222> <400> atg ott gtc ato att ttg ato stt gta ots get agt otg ace gtg Log Net Lou Val Ile Ile Lou Ile Iie Val Lou Ala So: Lou Thr Val Tb: sta att tot tao cor aaa stg Log gas tta aca aag Ile Ile Se: Tyr Pro Lys Met Thr Glu Lou Tb: Lye tee gtt 9am ass Ser Val Giu Lye ca Ctt gas Gin Lou Glu gat sat got gat nat ota tos gao ca ctg Los tat cag Asp Asn Ala Asp Asn Lou So: Asp Gin Lou Thr Tyr Gin ata gas Ile Glu gtg gog ca aaa Val Ala Gln Lys can ato tao gtg act aat- cag ota aac Gin Ile Tyr Val Thr Ann GlnLeu.-Ann gt stg 055 Arg Met Gin oag gaa stt Gin Glu Ile 70 etc sgt ago tts le Sex Arg Lou ccg, ata Pro. 110'; 75 tgc gt- cag s at Cys Val Gin"fAsf ass te ga tta acq gag sgt oga gat Lye So: Ala Lou Thr Glu Sex flg Asp tos -gac aaa. oga Ser Asp Lye Arg ttg gaa Lau Glu ttg att sac Lou Ile Amn sat tta tot cag tca gtt cag aaa atg caa get tos Asn Lou Sor Gin So: Val Gin Lys Met Gin Asp So: stg aaa apo got tgg ate aaa Met Lye Agn Ala Trp Ile Lys 115 gao aaa ctg ttg agg aa ago tgg Ala Lye Lou Lou Arg Lys Sc: Trp 125 aaa ass Lye Lys 130 Cgo tao aaa ago Axg Tyr Lye Arg goa aae tto ttt Ala Asn Phe Phe act gta tog ogt Thr Val So: Arg ota gag ago gto Leu Giu Be: Val ora ggt otg ggt Gin Gly Lou Gly tgg &as otg tgo Trp Lye Lou Cys gat gtt ggt ace act gaa aaa agt ctg Asp Val Sly Thr Thr Giu Gin Sec Lou 165 taa eat Se Asn 170 act sag ace Thr Lys Thr cgg ggs Arg Sly 175 ate tta ggg Ile Lou Sly gag ttc Glu Lou 180 caa atc gqt coa stt ats gac Sin Lou Sly Gin Ile Ile Glu 185 gat att atg aos Asp lie Met Thr 190 gtt egt caa tat gag aga gas ttt cct acg gtg tot ggc tot tat gag val Sor Gin Tyr Glu Arg Giu Phe Pro Thr val Sot Gly Sor So: Glu 195 200 205 cgt gtt gas tat gat att aaa tac ctg gas atg gte egg gag att sta Arg Val Glu Tyr Ala Ile Lys Tyr Lou Glu Met Val Arg Glu Ile Ile 210 215 220 tot stt tgc eta ttg act eta sgt tta tot aga ags tts tte Ser Ile 225 Cys Lou Leu Lou Sot Ph. So: Arg Arg Lou Lou 235 ttc aaa tgg aac Ph. Lys Trp Amn cog att Pro I1e 240 tet stt Set II 255 624 672 720 768

816- 864 ggg aga tgc tts Gly Arg Cys Lou sat tgg gtg sce Asn Trp Val Thr cgt, ant ctt Arg Asn Lou &at sag tac Asn Lys Tyr 275 tac tgg Tyr Trp 260 gcs agt stt cgt aaa ttt gee Ala 5cr-le Lys Pho Ala aa gat eta aae- Lyi Asp Ile Asn 270 tts at cot act Lou Awn Pro Pro gas (lu 280 acg ace sat ttt Thr Thr Asn Pho ,ggt sly 285 ate 'atg ttcl. le met Pho tts cca Lou Pro 290 act gas ggg cta Thr Glu Gly Lou tat tct Tyr Sot 295 gas gtg gt Slu Val Val sat gca aca tto Awn Ala Thr Phe ttt gst agt eta Pho Asp So: Lou 305 Aga cgt gac gaa sat stt gte Ar; Arg Asp Glu ^an Ile Val 310 315 gts got gga cog Val Ala Sly Pro tea Sa 320 scc tte tot got tao taA Thr Lou Sor Ala Tyr 325 <210> 21 <211> 325 <212> PRT <213> group <400> 21 Met Lou Val B streptococcus Ile Ile Lou Ile Ile Val Leu Ala Ser Leu Thr Val Thr Ile Ile Sar Tyr Pro Lye Met Thr Glu Lou Thr Lye Sot Val Glu Lys Gin Lou Glu Asp Asn Ala Asp Ann Lou So: Asp Gin Lau Thr Tyr Gin 40 Ile Arg Lye Lau met Lye Gln 145 Asp Ile Val Glu et Sec Ile Lys Lye 130 Lou Val Lou Se: Val Gin Ala Asn Ann 115 Arg Glu Gly Sly Gin Ala Gin Lou Bar Soc 100 Ai~k Tyr Sc Th 6hz 180 Tyr I Gin Glu *Thc Asn Trp Lye Val Thr 165 Lou Glu Lys Ile 70 GU G1u Lou lle Arg Ann 150 Glu Gin Mrg Asp Sec Lye Val 135 Gin Gin Lou Glu Lye 215 Lou Gin Ile Tyr Val Thr Ann Gin Lou Asn Sex Arg Gin cys 120 Ala Gly Ser Sly Phe 200 ryr 3er SArg Asp Sar 105 Ala Asn Lou Lou Gin 185 Pro Lou Phe Lou Pro Ile 75 Arg Ber Asp 90 Val Gin Lye Lye Lu.,Lou Phe Phe Glu 140 Sly Arg Yrp 155 So: Asn Thr 170 Ile Iio.Glu Thr Val Sot Glu Met Val 220 3cr Arg Ag 235 Mrg Phe Lye 250 Lye Ph. Ala Thr Ann Pho I Cys Val Lye Arg Met Gin 110. Arg Lye'- 125 Thr. Val Lye LOu Lye Tb: Asp Ile 190 Giy 8cr 205 Arg Glu Lou Lou Trp Asn Lys Asp 270 fly Ile e 285 Gin Lou Asp Bar- Cys Arg 175 Met Sex Ile Pro Sot 255 Ile Met Asn Glu Se: Trp. Ar; ln 160 Gly Th Glu lie Ile 240 Ile Asn Phe 195 225 230 Gly Arg Ann Mrg Cys Asn Lou Lye Tyr 275 Lou Trp 245 Tyr Trp 260 Lou Asn rrp Val Ser Ile Pro Glu 280 23 Leu Pro Thr Glu Gly Lou Tyr Sot Glu Val Val Arg Asn Ala Thr Ph. 290 295 300 Phe Asp 5cr Lou Arg Arg Asp Glu Ann Ile Val Val Al. ely Pro Ser 305 310 315 320 Thr Lou er Ala Tyr 325 <210> 22 <211> 579 <212> DNA <213> group B streptococcus <220> <221> CDS <222> <400> 22 at; ega aa gaa gtg ace coa gag at; ctt aac tat net Arg Lys Glu Val Thr Pro Glu Mot Lou Ann Tyr 1 5 ggc Sly gsa Glu ttt Ph. 990 Gly gct Ala ttt att Phe le ett gtt Len Val cgt ttt Arg Phe ggt aac Sly Asn att aga Ile Arg gaa Glu gaa Glu att Ile ttg Leu age Sot ate Ile tca Ser tt. Lou cta Lou att Ile gtt aa Val Lys got ttt Ala Phe aaa tat Lys Tyr aga ggc Arg Sly tca cgt Sr Arg att aaa gag tat tgt aac ttt ggt tgt get tat ttt gtg ttg gag sat Ile Lys Glu Tyr Cys Asn Phe Cys Ala Tyr Phe Val Lem Glu Asn cca aat Pro Asn cgt. ag Arg Lys 130 aat cgt Ran Arg 145 aaa gac Lye Asp sag gas Lys Glu taa 100 cet aga gat att aaa Pro Arg Asp Ile Lys 115 tea aga tee aaa tea Ser Arg Ser Lys Ser 135 tee cag tea aat gcc Ser Gln Se Asn Ala 150 aca aa cge cgt caa Thr Lys Arg Arg Gln 165 atg ace tot geea ag Met Thr Ser Ala Lys 180 105 get gat gas Asp Asp, Glu tea tea aag ser Ser Lys get cat ttt Ala His Phe 155 cgt cat att Arg His lie .170 cat ttg tta His Lau Leu 185 110 cct cat Pro His 125 caa act Gln Thr agt aaa SeX Lys gaa gag GlU Glu gta aga Val Arg 190 aag cgt Lys Arg aga aat Arg Asn sag cgt Lys Arg 160 caa gat. Gin Asp 175 aaa sat Lys An 384 432 480 528 576 <210> 23 <211> 192 <212> PRT <213> group B streptococcus <400> 23 Met Arg Lys Glu Val Thr Pro Glui Met 1 5 Gly Pro Gln Phe 1le His Phe Glu Ann 25 Gl Phe Gln Lou Val Ile Asn Glu Lys 40 Phe Gly Gln Arg Phe Ser Glu Ile Leu 55 Gly Asp Trp Gly Asn Glu Gin Leu Arg 70 Ala Ser Thr Ile Arg Lys Asn Ser Arg Leu An Tyr Asn Lys 10 1ie Val Lys Ser Asp Ser Ala Phe Asp Val Leu Lys Tyr Asp Phe Leu Arg Gly Phe Tyr 75 Ile Ser Arg Leu Glu Tyr Pro Asp Ile Thr Val Ile Val Lys Asp Asp Tyr Ile Pro Arg Asn 145 Lys Lys Lys Glu Tyr 100 Asa Pro Arg 115 Lys Ser Arg 130 Arg Set Gin Asp Thr Lys Glu Met Thr 180 Cys Asn Asp Ile Ser Lys Ser Asn 150 Arg Arg 165 Ser Ala 90 Gly Cys Ala 105 Phe Asp Asp 120 Gin Ser Ser Asa Ala His Glu Arg His 170 Gin His Leu .185 Val Leu 110 Pro His 125 Gin Thr set Lys Glu Glu Val Arg 190 Glu Asn Lys Arg Arg Asn Lys Arg 160 Gln Asp 175 Lys Asn-- <210> 24 <211> 609 <212> DNA <213> group B streptococcus <220> <221> CDS <222> <400> 24 atg aca ata aaa aaa gtg tta Met Thr Ile Lys Lys Val Leu 1 gta gcg tct cta get get tgt Val Ala Ser Leu Ala Ala Cys ggc aaa aaa gaa gtt aat ttt Gly Lys Lys Glu Val Ash Phe tct tat gtg sag gat ggg aaa Ser Tyr Val Lys Asp Gly Lys 55 agt gta Ser Val age tea Ser Ser 25 gca act Ala Thr 40 ctg act Leu Thr gga att att tta Gly Ile l1e Leu tct cat act act Ser His Thr Thr gga aca acg gca Gly Thr Thr Ala ttt gat att gaa Phe Asp Ile Glu aca Thr acg Thr ttt Phe gce Ala aaa get gtt ttt aaa ggt tca gat aac tat Lys Ala val Phe Lys Gly Ser Asp Ann Tyr gtc act ttt aaa Val Thr Phe Lys aca gaa tgg Thr Glu Trp atg ggt gga Met Gly Gly tta ttt tca Leu Phe Ser 115 tea teg Ser Ser aat aat Asn Ann 100 gta ttt acc ggc Val Phe Thr Gly get tca gga aag ttt caa Asp Ser Gly Lys Phe Gin att tct tat tea tca gag aga tot eaa aaa tay Ile Ser Tyr Ser Set Glu Arg Ser Gln Lys Tyr tac cca ata gg99 Tyr Pro Ile Gly tct act cot tea gtt tta gca gtt cct ser Thr Pro Ser Val Leu Ala Val Pro 120 125 aag aat agt sat atc aaa get Lys Asn' Ser Asn 1le Lys Ala 130 135 tat sat gat att Tyr Asn Asp Ies agt ggt,, cat aaa aca Set Gly His Lys Thr 140 cag gtt Gin Val 145 gt caa gga Val Gin Gly aca act geec aag Thr Thr Ala Lys tta gaa aat ttc Leu Glu Asn Phe aaa gag cat cag Lys Glu His Gin aat cct gtt act Ann Pro Val Thr aaa tat act aat Lys Tyr Thr Asn gaa aat Glu Asn 175 att aca cag att eta acg aat ttg Ile Thr Gln Ile Leu Thr Asn Leu 180 get gga aaa get Asp Gly Lys Ala get ttt aa Asp Phe Lys 190 ctt ttg acg Leu Leu Thr 195 gac caa ctg tta Asp Gln Leu Leu acg eta tta tea Thr Leu Leu 200 <210> <211> 202 <212> PRT <213> group B streptococcus <400> Met Thr Ile Lys Lys Val Leu Ser Val Thr Gly Ile Ile Lou Val Thr Val Ala Ser Leu Ala Ala Cys Ser Ser Lys Ser His Thr 25 Thr Lys Thr Gly Lys Lys Glu Val Asn Phe Ala Thr Val Gly Thr Thr Ala Pro Phe set Tyr Val Lys Lys Ala Val Phe Thr Glu Trp Soe Met Gly Gly Asn 100 Leu Phe Sex Tyr 115 Lys Asn ser Asn 130 G1n Val Val Gln 145 Lys Glu His Gin Ile Thr Gin Ile 180 Leou Leu Thr Asp 195 Asp Lys Set Asn Pro. Ile Gly Lys 165 Leu Gin Gly Lys Gly Ser 70 Val Phe Ile Sex Ile Gly Lys Ala 135 Thr Thr 150 Asn Pro Thr Asn Leu Leu 40 Leu Thr Asp Asn Thr Gly Tyr Ser 105 Ser Thr 120 Tyr Asn Thr Ala Val Thr Leu Ser 185 Thr Leu 200 Gly Tyr Ile 90 Ser Pro Asp Lys Leou 170 Asp Leu lie Glu Thr Phe Gly Lys Ser Gin 110 Leu Ala 125., 'Gly, Hid Glu Asn Thr Asn Ala Asp 190 Val Ala Lys Lys Phe Gin Lys Tyr Val -Pro 'Lys Thr Phe Asn 160 Glu Asn 175 Phe Lys <210> 26 <211> 357 <212> DNA <213> group B streptococcus <220> <221> CDS <222> <400> 26 atg aag aat ata aca aag cta tea act gtt got tta agoe tata a ctt 48 Met Lys Asn Ile Thr Lys Leu Sex Thr Val Ala. Leu Ser Leu Leu Leu 28 I tgt acg gcg Cys Thr Ala cat cat cct His His Pro get gca tca aac Ala Ala Ser Asn acg tot aca tct aaa aca cag tct Thr Ser Thr Ser Lys Thr Gin Ser aca gat aag caa aaa gaa gaa ccc Thr Asp Lys Gln Lys Glu Glu Pro aaa caa act Lys Gln Thr aaa atc Lys Leu aaa aac Lys Asn aa gaa get get Lys Glu Ala'Ala caa gag atg cat cc caa gge get gtt 192 Gin Gl Met His Pro Gin Gly Ala Val ttg aca aaa tat Leu Thr Lys Tyr gca aaa ccg gtc Ala Lys Pro Val aaa gat Lys Asp tAt gga aha Tyr Gly Lys anaa 240 Lys ate gat gtt ggt gat ggc aag aaa atg Ile Asp Val Gly Asp Gly Lys Lys Met 86 att tat gaa act Ile Tyr Glu Thr ggt cag 288 Gly Gin gga aaa att Gly Lys 1le get atg ctt Ala Met Leu 115 att gtt ttt att lie Val Phe Ile ggt caa get gag Gly Gln Ala Glu att cge cac Ile Arg His 110 ata aga att tea Ile Arg Ile B streptococcus <210> <211> <212> <213> 27 118 PRT group <400> 27 Met Lys Asn 1 Cys Thr Ala Ile Thr Lys Leu Ser Thr Val Ala 5 10 Leu Ser Leu Leu Leu Cys Ala Ala Ser Asn Thr 25 Ser Thr Ser Lys Thr Gln Ser His His Pro Lys Gln Thr Lys Leu Thr Asp Lys Gln Lys Glu Glu Pro Lys Asn Lys Glu Ala Ala Asp Gin Glu 55 Met His Pro Gin Gly Ala Val Asp Leu Thr Lys Tyr Lys Ala Lys Pro Val Lye Asp Tyr Gly Lys Lys 70 75 Ile Asp Val Gly Asp Gly Lys Lys Met Asn Ile Tyr Glu Thr Gly Gin 90 Gly Lys le Pro Ile Val Phe Ile Pro Gly Gln Ala Glu Ile Arg His 100 105 110 Ala Met Leu Ile Arg Ile 115 <210> 28 <211> 1191 <212> DNa <213> group B streptococcus <220> <221> CDS <222> (1)..(1191) <400> 28 gtg ast gaa tcg acc ate aga aaa Val Asn Glu Ser Thr Ile Arg Lys 1 5 tgg ate tta oat aat caa gca gtt Trp Ile Leu Asn Asn Gln Ala Val gta ttt tta acg att ttt att ttt Val Phe Leu Thr Ile Phe Ile Phe 40 cct gtg ttt gat ttt ctt get gtg Pro Val Phe Asp Phe Leu Ala Val 55 ggc ttg ctt tat tac eta tta aaa Gly Leu Leu Tyr Tyr Leu Leu Lys 70 cgg gga att aag cgt gta aca gcg Arg Gly Ile Lys Arg val Thr Ala gas ttt aaa ata Glu Phe Lys Ile 10 att gct ctc atg Ile Ala Lou Met 25 acc asa atc tot Thr Lys Ile Ser ctg ata ttg cog Leu Ile Leu Pro cct atg gtt aca Pro Met Val Thr ata tta tca gtt Ile Leu Ser Val 90 gtt Val att Ile ttt Phe ctt Leu ttt Phe ttt Phe gtt ttt Val Phe acc ttt Thr Phe atg ttt Met Phe gta att Val Ile tta gag Leu Glu act att Thr Ile ata ctt ctg tta att tgg gca at; tat apt ttt att ccc at; at; apt Ile Len Leu Leu Ile Trp Ala Met Sec set PMe Ile Pro Met Net Set aat ceaa tta cga cat ttt atg Asn Gin Le 115 gtp caa at; Val Gin Met 130 Arg His Phe Met gas pat etc cat tca tat gtg ant aaa Glu Asp Len Pro Ser Tyr Val Ann Lys 120 125 gin. aea apt Gin Thr Ser ttt atea gat cam Phe Ile Asp His ect tgg tta aaa Pro Trp Len Lys tat aaa gpg gac Tyr Lys ely Glu tcg aga atg tta Sor 5cr Met Lem tat act ate agt 8cr An Ile Sex 155 age cam 8cr Gln 160 gap qta tat tat pat Ale Val Scr Tyr Ale 165 gaa aaa ttt te Gin Lys Phe Sec aap aat qtt Lye Axn-Val 170 tta gat tgp pea Leu Asp Trp Ala 175 gga eat tta Giy An Leu gct ccc ttt Ala Pro Phe 195 gct agt Ala 8cr 180 ina gtt gc cgt gtg aca gta gca Thr Val Ala Arg val Thr Val Aln 185 ace atc at; Thr Ile Met 190 mac atg nag Asn Met Lys att ttp ttt tat Ile Lem Phe Tyr att Leu 200 tta aga pat egt Leu Arg Asp Set eat ggt Asn Gly 210 ttc tta at; gtt Phe Lem Met Val tta Lem 215 cca acc aaa cta Pro Thr Lys Lem cpa can ca gt Arg Gin Pro Ala 220 gm tat gtg can Sly Tyr Val Gln get Asp gga ly 240 att ttg cga ga at; Ile Len Arg Glu Met 230 aat mgt cam at; Asn ser Gin Met te Ser 235 672 720 768 cam atc att gtt pat Gin lie Ile Vel Aim 245 mtt tat gtt ;gt Ile Thr Val Gly ntt ttt tca ate Ile Phe Ser Ile at; tat Met Tyr 255 apt mtt eta Ser Ile Ile ggC ctt aga tat ggc gtg Sly Leu Arg Tyr Sly Val 260 265 aca tta ggp att Thr Leu Gly Ile ctt gao ggt Ile Ala Sly 270 ptg tta mat atg gtt ceo tat ttg Val Lem Asn Met Vel Pro Tyr Leu 275 280 gga apt ttt gta gca cam att cca Sly Sec Phe Val Ala Gin lie Pro 285 gtg ttt ata tta gag ctt gtc 9ca gga cat gtt atg gtt gtt aaa gtt Val Phe lie Leu Ala Lou Val Ala Gly Pro Val Met Val Val Lye Val gg Ala 305 att gtt ttt gtt lie Val Phe Val gag ca act eta gag 9ga cgC ttt gtc tca Giu Gin Thr Leu Glu Sly Arg Phe Val Sex cot ttg gtt tta Pro Lou Val Leu ggt mat Sly ABn 325 aaa ctt ago Lys Lou Ser cat cea att ace His Pro Ile Thr ntt etg lie Net 335 1009 1056 ttt att ttn Phe lie Leu otc agt att Lou 5cr Ile 355 tta Lou 340 ace tat gga 909 Thr Ser Siy Ala ttt ggt gtt tgg gma gte tto Phe Sly Val Trp Sly Vml Phe 350 Cog att tat 9ca Pro Il eTyr Ala tat Ser 360 ata aa gtt gtt Ile Lye Val Val gtt ama Oaa ttg Val Ly. (lu Lou 365 ate gat gtt gtt le Asp Val Val 1104 1152 ttt gut Ph. Asp 370 tgg tao ama get Trp Tyr Lye Ala ngt ggg ata tat Sex Gly Lou Tyr act gaa gaa agn agt gaa ga gtt aaa nat gtt gas tag Thr Giu (lu Arg So: Glu Glu Val Lye Ann Val Glu 1191 <210> 29 <211> 396 <212> PRT <213> group <400> 29 Val Ann Glu 1 Trp Ile Leu B streptococcus Sex Thr Ii Arg Lye Clu 5 Asn As Gin Ala Val Ile 25 Lys Ile Val Val Phe Lye Ala Leu Met lie Thr Phe Leu Met Phe Lys Val Phe Lou Thr lie Phe Ile Thr Lys Ile Ber Pro Val Ph. Asp Ph. Lou Ala Val Lou lie Leu Pro Lou Val Il eer Gly Leu Leu Arg Gly Iie Ile Lau Lou Asn Gln Leu 115 Val Geln Met 130 ser Tyr Lys 145 Ala Val Set Gly Awn Leu Ala Pro Phe 195 Asn Gly Phe 210 Arg Ile Leu 225 Gin Ile 1e Ser Ile le Val Leu Asn 275 Val Phe Ile 290 Tyr Lys Leu 100 Arg Glu Gly Tyr Ala 180 Ile Leu Arg Val Gly 260 Met Leu Ty SAr His Thr Glu Ala 165 Set Leu Met Glu Ala 245 Leu Val Ala r Leu 70 g Val Trp a Phe Ser I lie0 150 Glu Thr Phe Val Met 230 Ile Arg Pro Leu Ile 310 a Leu Lys Pro Thr Ala Ile Ala Met Set 105 Met Giu Asp 120 Ser Phe Ile 135 Ser Set Met Lys Phe Ser Val Ala Arg 185 Tyr Leu Leu 200 Leu Pro Thr 215 Asn Set Gin Thr Val Gly Tyr Gly Val 265 Tyr Leu Gly 280 Val Ala Gly 295 Glu Gin Thr Met Leu 90 Set Leu Asp Leu Lye 170 Val Arg Lys Met Val 250 Thr Set Pro Leu Val Thr 75 a Ser Val Phe Ile Pro Set His Asn 140 ser Asn 155 Aan Val Thr Val Asp Ser Leu Arg 220 ser Gly 235 Ile Phe Leu Gly Phe Val Val Met 300 Glu Gly 315 Phe Phe Pro Tyr 125 Pro Ile Lou Ala Arg 205 Gln Tyr Ser Ile Ala 285 Val A.rg Leu Thr Met 110 Val Trp Ser Asp Thr 190 Aan Pro Val Ile Ile 270 Gln Val Phe Glu Ile Met Asn Leou Ser Trp 175 Ile Met Ala Gin Met 255 Ala Iie Lys Val Lys Ile Set Lys Lys Gin 160 Ala Met Lys Asp Gly 240 Tyr Gly Pro Val Set 320 Ile Val Phe Val Pro Phe Leu Phe Thr 385 Val Leu Leu Lou 340 Ile Pro 355 Trp Tyr Glu Arg Gly Asn 325 Thr Set Ile Tyr Lys Ala Ser Glu 390 Lys Leu Gly Ala Ala Ser 360 Val Sex 375 Glu Val Ser Ile His Pro Ile Thr Ile Met 330 335 Met Phe Gly Val Trp Gly Val Phe 345 350 Ile Lys Val Val Val Lys Glu Leu 365 Gly Leu Tyr Thr Il Asp Val Val 380 Lys Asn Val Glu 395 <210> <211> 1230 <212> DNA <213> group B streptococcus <220> <221> CDS <222> (1)..(1230) <400> atg ttt atg gga ate cca caa Met Phe Met Gly Ile Pro Gin 1 5 eta cca att tac ate ggc tta Leu Pro Ile Tyr Ile Gly Leu gag att att ttt agt eta agt Glu Ile Ile Phe Ser Leu Ser act ttt aat caa ttg aag tca Thr Phe Asn Gln Leu Lys Ser 55 ctg eta gtt ttt ate tat aaa Leu Leu Val Phe Ile Tyr Lys 70 ttg gtC ttt tat gta acg gtt tat Tyr tte Phe ttt Phe 40 cta Leu get Ala tgt 34 ttc ttc tac Phe Phe Tyr ttt aag aag Phe Lys Lys 25 att gta atg Ile Val Met ttg gca tac Leu Ala Tyr tac egg aaa Tyr Arg Lys tta tct att ctt ate tta Leu Ile Leu cgt ttt gac Arg Phe Ala atg ttg act Met Leu Thr gtt gtc gga Val Val Gly cga ttt aat Arg Phe Ann ttt cog eta 48 96 144 192 240 288 Lou Vel Phe Tyr Vol Thr Val Cys Lou Ser Ile Ph. Pro Lou Ph. Lou gt aaa tte Val Lye Lou ggg ttt cta ely Ph. Lou 115 cca got eta tot gag get ggg cat cag Pro Ala Ile Sec Glu Asp ely His Gin 105 toa ctt ttt Sor Lou Pho 110 gga ett tot tao Gly Ile Ser Tyr act ttt age got gtt got etg ott Thr Phe Arg Ala Val Ala Met fle 125 att gaa Ile Glu 130 ctg ega gao Nt Arg Asp ggt gtc ely Val 135 ttg aaa gaa ttt act Lou Lye Glu Phe Thr 140 ttc tgg gac tta Lou Trp ClU Ph. 432 .480 aga ttt tta otc Arg Phe Lou Lou ttt cc& act tto Ph. Pro Thr Phe too agt 1er 5.r 255. gga cea att ely Pro noe agt ttt aaa aga Arg Phe Lys Arg ttc aat Phe Axn 165 gag get tao Glu Asp Tyr aat ate cc gat Asn Ile Pro.,Asp Ogo&q sac 52 Arg Ann 175 gam otc ota Clu Lou Lou ttt cto tat Phe Lou Tyr 195 ctg tta ggt caa Not Lou ely Gln ett oat tat Ile His Tyr ttg ctg tte ggt Lou met Lou Gly 190 576 624 eag ttt ett tte Lye Phe Ile Lou tat att ttt ggo Tyr Ile Pho Gly atg att etg Lou le Met cot cat Pro Pro 210 ota aae gee ttc Lou Lye Glu Lou gcg Ale 215 ota gaa cog ggt Leu Glu Gin Sly gtg ttt at tgg Vel Pho Asn Trp aca ott ggg gtt Thr Lou Gly Val tat gao ttt ggt Tyr Ala Phe Gly ttt gut Pho Asp 235 ttg ttc ttt Lou Phe Phe ttt gca ggt tac Phe Ala ely Tyr etg ttt gag Met Phe Ala ttg get Lou Ala 250 ott tot acc ota Ile Ser Asn Lou etg ggg Met Sly 255 ott aeg tct le Lys Ser ett eac ttt gao Ile Asn Phe Asp aaa cot tto tea ogo gac cta Pro Ph. Lys Soc Arg Asp Lou 270 aaa gaa ttt tgg cat aga tgg cat atg ago ctt tot ttc tgg ttt aga Lys Glu Phe Trp Asn Arg Trp His Met Sor Lou Ser Phe Trp Phe Arg 275 280 285 gac ttt Asp Phe 290 gtt ttc atg egg Val Ph. Met Arg gtt ang ctt tta gtt maa aat Val Lye Lou Lou Val Lys Ans 300 ttt ama eac cgt eat gtt act te Phe Lys Asn Arg Asn Val Thr Ser 305 310 gte get tat att ate Val Ala 315 Tyr Ile le aaa gtt Lye Val eat atg Asn Met 320 goc tat Ala Tyr 335 ctt tt atg ggm Leu Lou Met dly tgg cat gg tta trp His Giy Leu tgg tao tat ate ?rp Tyr Tyr Ile 1008 ggt ato ttt Sly Lou Phe nag mag aaa Lye Lys Lye 355 cat ggg His Gly 340 mtt ggc cta Ile Sly Leu mtt eat ga' got Ile Asn Asp Ala tgg gte cgt Trp Val Arg' 350 aaa ocm ott Lys Pro Lou 1056 1104 mat ayt mat Asn Xaa Asn Aaa gaa Lys Glu 360 .aga aga ttg get Arg Arg Lou Ala tta cca Leu Pro 3'70 gaa aac ama tgg Glu Asn Lys Trp tat got ttg ggt Tyr Ale Lou Gly ttc ate acc ttt Phe Ile Thr Phe 1152' gte gtt tg ttt Val Val Met Ph. ttc ttg att ttt tca ggm ttt tta gat ott Ph. Lou Ile Phe Ser Sly Phe Lou Asp Leu 395 400 1200 ttg tg tto cca Leu ?rp'Phs Pro cam cg Gin Pro 405 oat aac aaa tam His Ann Lye 1230 <210> 31 <211> 409 <212> PRT <213> group 5 streptococcus <400> 31 Met Phe Met Gly lie Pro Gin 1 5 Leu Pro Ile Tyr Ile Gly Lou Tyr Phe Ph. Tyr Lou Ile Leu Ala Val 10 Phe Phe Lys Lys Arg Phe Ala Lou Tyr 25 Glu Ile Ile Phe Ser Leu Ser Phe Ile Val Met Met Leu Thr Gly Set 40 Thr Leu Leu Val Gly Ile Leu 145 Arg Glu Phe Pro Pro 225 Phe Ile Phe Asn Leu Val Val Phe LyS Leu Phe Leu 115 Glu Met 130. Arg Phe Phe Lys Leu Leu Leu Tyr 195 Pro Leu 210 Thr Leu Ala Gly Lys Ser Gln Phe Tyr Ile 100 Gly Arg Leu Arg Asp 180 Lys Lys Gly Tyr Pro Leu Ile Val Pro le Asp Leu Phe 165 Met Phe Glu Val Thr 245 Ile Lys Tyr 70 Thr Ala Ser Gly Phe 150 An Leu Ile Leu Met 230 Met Asn Ser 55 Lys Val Ile Tyr Val •135 Phe Glu Gly Leu Ala 215 Tyr Phe Phe Leu Ala Cys Set Leu 120 Leu Pro Asp Gln Ala 200 Leu Ala Ala Asp ILeu Ala Tyr Arg Leu Ser 90 Glu Asp 105 Thr Phe Lys Glu Thr Phe Tyr Ile 170 Ala Ile 185 Tyr Ile Glu Gin Phe Gly Leou Ala 250 Lys Pro 265 Tyr Lys 75 Ile Gly Arg Phe Ser 155 Aan His Phe Gly Phe 235 Ile Phe Val Arg Phe His Ala Thr 140 Ser Ile Tyr Gly Gly 220 Asp Ser Lys Val Phe Pro Gin Val 125 Leu Gly Pro Leu Ser 205 Val Leu Asn Sex Gly Asn Leu Sex 110 Ala Tkp Pro Asp Met 190 Leu Phe Phe Leu Arg 270 Gln Set His Thr Phe Lou Leu Phe Met' 'Ile Glu Ph Ile Asp 160 Arg Asn 175 Leu Gly Ile Net Asn Trp Phe Asp 240 Met Gly 255 Asp Leu 260 Lys Glu Phe 275 Trp Asn Arg Trp Met Ser Leu Ser Trp Phe Arg Asp Phe Val 290 "Phe Lys Asn 305 Leu Leu Met Gly Leu Phe Lys Lys Lys 355 Leu Pro Glu 370 Asn Val Val 385 Leu Trp Phe Phe Met Arg Asa Gly Phe 325 His Gly 340 Asn Xaa Asn Lys Met Phe Pro Gln 405 Leu Val Lys 295 Thr Ser Ser His Gly Lou Gly Leu Val 345 Lys Glu Arg 360 Thr Tyr Ala 375 Phe Leu Ile His Asn Lys Leu Val Lys Asn Lys Val 300 Ala Tyr lie Ile Asn Met 315 320 Trp Tyr Tyr Ile Ala Tyr 335 Asn Asp Ala Trp Val Arg 350 Leu Ala Lys Lys Pro Leu 365 Gly Val Phe' I-le Thr Phe 380 Set Gly' Phe'Lu Asp Leu 395 400 <210> 32 <211> 100 <212> DNA <213> group B streptococcus <220> <221> CDS <222> <400> 32 atg aat aaa ata acg aca tta tca acc ate gqc ctg act tta atg ctt Met Asn Lys Ile Thr Thr Leu Ser Thr Ile Ala Leu Thr Leu Met Leu 1 5 10 tgc gtt gga tgt tot gcc aat aaa gat aat caa aaa act aaa act gag Cys Val Gly Cys Ser Ala Asn Lys Asp Asn Gln Lys Thr Lys Thr Glu 25 gat c Asp <210> 33 <211> 33 <212> PRT <213> group B streptococcus <400> 33 Met Asn Lys fle Thr Thr Leu Ser Thr l2e Ala Leu Thr Leu Met Leu 1 5 10 Cys Val Gly Cys Ser Ala Asn Lys Asp Asnn Gln Lys Thr Lys Thr Glu 25 Asp <210> 34 <211> 654 <212> DNA <213> group B streptococcus <220> <221> CDS <222> <400> 34 gat cga ggc tat caa gaa gca atg gct aaa cta agg aaa act tac ggc Asp Arg Gly Tyr Gin Glu Ala Met Ala Lys Lou Arg Lys Thr Tyr Gly 1 5 10 gaa tat ggt tta ggg gtt tct aca gga tta gat tta cot gaa tca gaa Glu Tyr Gly Leu Gly Val Ser Thr'Gly Leu Asp Leu Pro Glu Ser Glu 25 ggt tat gta Cct gga aaa tac age tta gga aca act cta atg gaa tog Gly Tyr Val Pro Gly Lys Tyr Ser Leu Gly Thr Thr Leu Met Glu Ser 40 ttc ggt cag tat gat gcc tat aca cca atg caa ctt ggt cag tat ate Phe Gly Gln Tyr Asp Ala Tyr Thr Pro Met Gln Leu Gly Gln Tyr Ile 55 tea act att gcg aat aat ggg aat cgt tta goa cct cac gtg gtt tca Ser Thr Ile Ala Asn Asn Gly Asn Arg Leu Ala Pro His Val Val Ser 70 75 gat atc tat gaa ggg aat gat tot aat aag ttc gct caa ttg gtt cgt 39 48 96 144 192 240 288 Asp Ile Tyr Sit' Sly Asn Asp 3cr Ann Lys Ph. Ala Gin Leu Val Arg 90 tea into act Sex Ile flit ttm gee mtt Lou Ala le 115 ggc tat gom Gly Tyr Mla 130 cot ain Pro Lys 100 mom ota mat Thr Lau Asn ata gct ate tee Ile Mla Ile Ser mtt oa 9am ggt Il@ Gin Sit' Sly tat mac gtt gte Tyr Ann Val Val gat cam gag Asp Gin Sit' 110 mgt gm ingt Ser Sly Bet moo mtt agy Thr Ile Xaa act ggt mog Thr ely flit tom atg Bar met 135 agg ggg mat Arg Sly An gtg inca Vinl Thx 140 mom act ggt, ace Lys Thr Sly flit gaa acm ttt got Sit' Thr Phe Ala ama at atm Rat. gga, Lys Asn Ile Ann-Sly 155- OCR gtt tct Thx Val Sex &ma at* gom Lys Ile Ala aaa tee cat Lys Her His 195 moo tao Thx Tyr 165 ao tta ae got Aush Lou Ann Ala gee tat get Ala Tyr Asp gog gta mtg tint Ala Val Net Tyr cat gtt mom mot His Val Thr 'Put mot mat cot Thr- Ann Arg 175 gat aca mom Asp Thr Thx 190 tat mtt tom Tyr Ile Sex 528 576 624 cam ttm gtt Gin Lou val gom cgm Alm Arg 200 gat atg mtt gat Asp Net Ile Asp Cag tom, cag gao mat mag age ggn cat tga Gin Sex Gin Asp Ann Lys Arg Siw His 210 215 <210> <211> 217 <212> PRT <213> group S streptococcus <400> Asp Arg Sly Tyx Gin Sit' Ala met Ala Lys Lou Axg Lys Thr Tr Sly (flu Tyx Sly Lou Sly Val Bar TI-a sly Lou Asp Lou Pro Sit' Sex Sit' V Gly Tyr Val Pro Gly b- 3 -Gln Tyr Asp Ser Thr Ile Ala Asn Asp Ile Tyr Glu Gly Sex Ile Thr Pro Lys 100 SLu Ala Ile Ile Gin 115 Gly Tyr Ala Thr Gly 130 Gly Lys Thr Gly Thr I 145 Thr Val Set Thr Tyr 165 Lys Ile Ala Val Ale 180 Lys Set His Gin Leu V 195 Gln Ser Gln Asp Asn L 210 Lys Tyr Ser Leu Gly Thr Thr 40 Ala Tyr Thr "Po Met 'Gln Lou 55 Asn Gly Asn Arg Leu Ala Pro 70 75 Asn Asp Ser Asn Lys Phe Ala 90 Thr Lou Asn Lys Ile Ala Il* 105 Glu Gly Phe Tyr Asn Vil Val 120 Thr Ser Met Arg Gly Asn Val. 135 140 Ala Glu Thr Phe Ala Lys Asn 150 155 sn Lou Asn Ala Ile Ala Tyr A 170 lal Met Tyr Pro His Val Thr T 185 al Ala Arg Asp Met Ile Asp G 200 2 ys Arg Gly His 215 Leu Met Gly Gln His Val Gin Leu Ser Asp 110 Asn Set 125 rhr Thr le Asn Lsp Thr 'hr. Asp 190 in Tyr I Gli Tyr Val Val Gln Gly Ile Gly Asn 175 rhr :le a Set Ile Set Arg Giu Set Xaa Gln 160 Arg Thr ser