Patent application title: Genes and proteins, and their use
Inventors:
Martin John Glenton Hughes (Berkshire, GB)
Joseph David Santangelo (Berkshire, GB)
Jonathan Douglas Lane (Berkshire, GB)
Robert Feldman (Berkshire, GB)
Joanne Christine Moore (Berkshire, GB)
Richard James Dobson (Berkshire, GB)
Paul Everest (Berkshire, GB)
Gordon Dougan (Berkshire, GB)
Rebecca Kerry Wilson (Berkshire, GB)
Assignees:
Emergent Product Development UK Limited
IPC8 Class: AA61K3909FI
USPC Class:
4241901
Class name: Antigen, epitope, or other immunospecific immunoeffector (e.g., immunospecific vaccine, immunospecific stimulator of cell-mediated immunity, immunospecific tolerogen, immunospecific immunosuppressor, etc.) amino acid sequence disclosed in whole or in part; or conjugate, complex, or fusion protein or fusion polypeptide including the same disclosed amino acid sequence derived from bacterium (e.g., mycoplasma, anaplasma, etc.)
Publication date: 2009-11-05
Patent application number: 20090274717
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Patent application title: Genes and proteins, and their use
Inventors:
Martin John Glenton Hughes
Joseph David Santangelo
Jonathan Douglas Lane
Robert Feldman
Joanne Christine Moore
Richard James Dobson
Paul Everest
Gordon Dougan
Rebecca Kerry Wilson
Agents:
Sterne, Kessler, Goldstein & Fox, P.L.L.C.
Assignees:
Emergent Product Development UK Limited
Origin: WASHINGTON, DC US
IPC8 Class: AA61K3909FI
USPC Class:
4241901
Patent application number: 20090274717
Abstract:
According to the present invention, a series of genes are identified in
Group B Streptococcus, the products of which may be associated with the
outer surface of the organism. The genes, or functional fragments
thereof, may be useful in the preparation of therapeutics, e.g. vaccines
to immunize a patient against microbial infection.Claims:
1-19. (canceled)
20. An isolated polypeptide comprising an amino acid sequence which has greater than 95% similarity to SEQ ID NO: 23.
21. The polypeptide of claim 20, wherein said amino acid sequence comprises SEQ ID NO: 23.
22. The polypeptide of claim 21, wherein said amino acid sequence is encoded by SEQ ID NO: 22
23. The polypeptide of claim 20, further comprising a heterologous polypeptide.
24. A composition comprising the polypeptide of claim 20 and a carrier.
25. The composition of claim 20, further comprising an adjuvant.
26. The composition of claim 25, wherein said adjuvant is alum.
27. The composition of claim 20, which induces an immune response against said polypeptide when administered in an animal.
28. The composition of claim 27, wherein said immune response is a humoral immune response.
29. The composition of claim 27, wherein said immune response is a cell mediated immune response.
Description:
FIELD OF THE INVENTION
[0001]This invention relates to the identification of bacterial genes and proteins, and their use. More particularly, it relates to their use in therapy, for immunisation and in screening for drugs.
BACKGROUND TO THE INVENTION
[0002]Group B Streptococcus (GBS), also known as Streptococcus agalactiae, is the causative agent of various conditions. In particular, GBS causes:
Early Onset Neonatal Infection.
[0003]This infection usually begins in utero and causes severe septicaemia and pneumonia in infants, which is lethal if untreated and even with treatment is associated with a 10-20% mortality rate.
Late Onset Neonatal Infection.
[0004]This infection occurs in the period shortly after birth until about 3 months of age. It causes a septicaemia, which is complicated by meningitis in 90% of cases. Other focal infections also occur including osteomyelitis, septic arthritis, abscesses and endopthalmitis.
Adult Infections.
[0005]These appear to be increasingly common and occur most frequently in women who have just delivered a baby, the elderly and the immunocompromised. They are characterised by septicaemia and focal infections including osteomyelitis, septic arthritis, abscesses and endopthalmitis.
Urinary Tract Infections.
[0006]GBS is a cause of urinary tract infections and in pregnancy accounts for about 10% of all infections.
Veterinary Infections.
[0007]GBS causes chronic mastitis in cows. This, in turn, leads to reduced milk production and is therefore of considerable economic importance.
[0008]GBS infections can be treated with antibiotics. However, immunisation is preferable. It is therefore desirable to develop an immunogen that could be used in a therapeutically-effective vaccine.
SUMMARY OF THE INVENTION
[0009]The present invention is based on the identification of a series of genes in GBS, and also related organisms, the products of which may be localised on the outer surface of the organism and therefore may be used as a target for immuno-therapy.
[0010]According to one aspect of the invention, a peptide is encoded by an operon including any of the genes identified herein as pho1-13, pho3-21, pho2-15, pho3-18, pho3-22, pho3-3, pho3-17, pho2-2, pho1-5, pho3-1, pho3-23, pho3-50, pho1-14, pho2-10, pho3-14, pho3-24 and pho3-29, obtainable from Group B Streptococcus, or a homologue or functional fragment thereof. Such a peptide is suitable for therapeutic use, e.g. when isolated.
[0011]The term "functional fragments" is used herein to define a part of the gene or peptide which retains the activity of the whole gene or peptide. For example, a functional fragment of the peptide may be used as an antigenic determinant, useful in a vaccine or in the production of antibodies.
[0012]A gene fragment may be used to encode the active peptide. Alternatively, the gene fragment may have utility in gene therapy, targeting the wild-type gene in vivo to exert a therapeutic effect.
[0013]A peptide according to the present invention may comprise any of the amino acid sequences identified herein as SEQ ID NOS. 2, 4, 6, 8, 10, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33 and 35, or a functional fragment thereof.
[0014]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 vaccines. For example, a vaccine may comprise a peptide according to the invention, or the means for its expression, for the treatment of infection. The vaccine may be administered to females prior to or during pregnancy to protect mother and neonate against infection by GBS.
[0015]According to another aspect of the invention, the peptides or genes may be used for screening potential antimicrobial drugs or for the detection of virulence.
[0016]A further aspect of this invention is the use of any of the products identified herein, for the treatment or prevention of a condition associated with infection by a Group B Streptococcal strain.
[0017]Although the protein has been described for use in the treatment of patients, veterinary uses of the products of the invention are also considered to be within the scope of the present invention. In particular, the peptides or the vaccines may be used in the treatment of chronic mastitis, especially in cows.
DESCRIPTION OF THE INVENTION
[0018]The present invention is described with reference to Group B Streptococcal strain M732. However, all the GBS strains and many other bacterial strains are likely to include related peptides or proteins having amino acid sequence homology with the peptide of M732. Organisms likely to contain the peptides include, but are not limited to, S. pneumoniae, S. pyogenes, S. suis, S. milleri, Group C and Group G Streptococci and Enterococci. Vaccines to each of these may be developed in the same way as described for GBS.
[0019]Preferably, the peptides that may be useful for the production of vaccines have greater than 40% sequence similarity with the peptides identified herein. More preferably, the peptides have greater than 60% sequence similarity. Most preferably, the peptides have greater than 80% sequence similarity, e.g. 95% similarity.
[0020]Having characterised a gene according to the invention, it is possible to use the gene sequence to establish homologies in other microorganisms. In this way it is possible to determine whether other microorganisms have similar outer surface products. Sequence homologies may be established by searching in existing databases, e.g. EMBL or Genbank.
[0021]Peptides or proteins according to the invention may be purified and isolated by methods known in the art. In particular, having identified the gene sequence, it will be possible to use recombinant techniques to express the genes in a suitable host. Active fragments and homologues can be identified and may be useful in therapy. For example, the peptides or their active fragments may be used as antigenic determinants in a vaccine, to elicit an immune response. They may also be used in the preparation of antibodies, for passive immunisation, or diagnostic applications. Suitable antibodies include monoclonal antibodies, or fragments thereof, including single chain fv fragments. Methods for the preparation of antibodies will be apparent to those skilled in the art.
[0022]The preparation of vaccines based on attenuated microorganisms is known to those skilled in the art. Vaccine compositions can be formulated with suitable carriers or adjuvants, e.g. alum, as necessary or desired, and used in therapy, to provide effective immunisation against Group B Streptococci or other related microorganisms. The preparation of vaccine formulations will be apparent to the skilled person.
[0023]More generally, and as is well known to those skilled in the art, a suitable amount of an active component of the invention can be selected, for therapeutic use, as can suitable carriers or excipients, and routes of administration. These factors will be chosen or determined according to known criteria such as the nature/severity of the condition to be treated, the type or health of the subject etc.
[0024]The products of the present invention were identified as follows:
[0025]A partial gene library of GBS (strain M732) chromosomal DNA was prepared using the plasmid vectors pFW-phoA1, pFW-phoA2 and pFW-phoA3 (Podbielski, A. et al. 1996. Gene 177:137-147). These plasmids possess a constitutive spectinomycin adenyltransferase antibiotic resistance marker, which confers a high level of spectinomycin resistance and is therefore easily selected. Furthermore, these vectors contain a truncated (leaderless) Escherichia coli phoA gene for alkaline phosphatase. The three vectors differ only with respect to the reading frame in which the leaderless phoA gene exists, as compared to an upstream in-frame BamHI restriction enzyme site. Because this truncated E. coli phoA gene lacks the appropriate leader sequence for export of this enzyme across the bacterial membrane, extracellular alkaline phosphatase activity is absent when these plasmids are propagated in an E. coli phoA mutant (e.g. strain DH5α). The chromogenic alkaline phosphatase substrate, XP (5-bromo-4-chloro-3-indolyl-phosphate), does not enter intact bacterial cells and therefore only exported or surface associated alkaline phosphatase activity can be detected. When exported or surface associated alkaline phosphatase activity is present, the chromogenic XP substrate is cleaved to yield a blue pigment and the corresponding bacterial colonies can be identified by their blue colour.
[0026]Plasmid DNA was digested to completion with BamHI and dephosphorylated using shrimp alkaline phosphatase. GBS genomic DNA was partially digested with Sau3AI, size fractionated on a sucrose gradient and fragments <1 kb in size were ligated into the prepared pFW-phoA vectors. E. coli strain DH5α was chosen as the cloning host since it lacks a functional phoA gene. Recombinant plasmids were selected on Luria agar containing 100 μg/ml of spectinomycin and 40 μg/ml of the chromogenic XP substrate. E. coli transformants harbouring plasmids containing GBS insert DNA that complements the export signal sequence of the leaderless phoA gene were identified by the blue colour of the colonies. Approximately 30000 different recombinant plasmids containing GBS insert DNA were screened in this manner and 83 recombinant plasmids, which complemented the leaderless phoA, were chosen for further study.
[0027]From these experiments, several clones were selected each containing a plasmid containing a gene (or part thereof), which complemented the leaderless phoA.
[0028]Having identified the gene in each clone it is then possible to obtain the full-length gene sequence, as follows.
[0029]Using the identified and sequenced gene fragment, oligonucleotide primers were designed for genomic DNA sequencing. These primers were designed so as to sequence in an `outward` direction from the obtained sequence. Once read, the sequence obtained was checked to see if the 5' and 3' termini of the gene had been reached. The presence of these features was identified by checking against homologous sequences, and for the 5' end the presence of an AUG start codon (or accepted equivalent) preceded by a Shine-Dalgarno consensus sequence, and for the 3' end, the presence of a translation termination (Stop) codon.
[0030]Upon identification of the full-length gene, primers were designed for amplification of full-length product. Primers used included restriction enzyme recognition sites (NcoI at the 5' end and EcoO109I at the 3' end) to allow subsequent cloning of the product into the Lactococcal expression system used.
[0031]PCR was carried out using the primers, and the products cloned into a pCR 2.1 cloning vector (In Vitrogen). Following confirmation of the presence of the cloned fragment, the DNA was excised using the restriction enzymes NcoI and EcoO109I.
[0032]The vector into which this fragment was inserted was a modified version of pNZ8048 (Kuipers, O. P. et al. (1998) J. Biotech 64: 15-21). This vector, harbouring a lactococcal origin of replication, a chloramphenicol resistance marker, an inducible nisin promoter and a multicloning site was altered by the replacement of the multicloning site with two 10× His tags, flanked on the 5-most end with an NcoI site, split in the middle with a multicloning site (including an EcoO109I site), and a Stop (termination) codon at the 3' end of the His tags.
[0033]The gene of interest was inserted so that a 10× His tag was in the 3' position relative to the coding region. Following transformation of the recombinant plasmid into L. lactis (strain NZ9000-Kuipers, O. P. et al. (1998) supra), a 400 ml liquid culture was set up and translation of the protein was induced by the addition of nisin to the culture. After a 2 hour incubation, the cells were harvested and lysed by bead beating. The resultant lysate was cleared by centrifugation, then passed over a metal affinity (Talon, Clonetech) column. The column was washed repeatedly before bound proteins were eluted with Imidazole.
[0034]To identify fractions containing the His-tagged recombinant protein, an aliquot from each fraction was analysed by SDS-PAGE, Western blotted and probed with anti-His antibodies.
[0035]The recombinant protein obtained was then used to immunize New Zealand white rabbits, with pre-immune sera being harvested prior to immunisation. Following a boost, the rabbits were sacrificed and sera collected. This sera was used in Western blots, ELISA and animal protection models.
[0036]Using the sera obtained from the animal studies, immunosorption studies were carried out.
[0037]Group B Streptococcus was grown in 20 ml Todd Hewitt broth (THB) for 8 hours, harvested and resuspended in 5 ml PBS. 50 μl aliquots of this were used to coat wells in a 96 well plate (Nunc Immuno-Sorb). This was left at 4° C. overnight to allow for absorbance of the bacteria onto the plate. Plates were washed twice with PBS, then blocked with 3% BSA in PBS for 1 hr at 37° C. Plates were again washed. Serial 10 fold dilutions of the sera were made in PBS and 50 μl of these dilutions were added to the wells of the plate, in duplicate. The plate was covered and incubated for 1 hr at 37° C. The plate was washed, then 50 μl anti-rabbit alkaline phosphatase conjugated secondary antibody at a concentration of 1:5000 was added to each well. Following incubation at 37° C. for an hour, the plate was washed again. 50 μl substrate (PNPP) was added to each well, and the reaction allowed to proceed for 30 min before the absorbance was read at 405 nm.
[0038]Animal protection studies were also carried out to test the effectiveness of protection on the immunised rabbits.
[0039]GBS M732 was grown up in THB until mid-log phase was reached--approximately 5 hours. Cells were counted in a counting chamber, and bacteria were diluted to give a concentration of 2×107 bacteria per ml in pre-immune or test sera. 50 μl of this was injected via the intraperitoneal route into 0-1 day old mice. The mice were observed for survival over 48 hours.
[0040]The following Examples illustrate the invention.
EXAMPLE 1
[0041]A first clone contained a gene sequence identified herein as SEQ ID NO. 1, with an amino acid sequence identified as SEQ ID NO. 2, and classified as pho1-13.
[0042]A comparison of the amino acid sequence of pho1-13 was performed.
[0043]Homologues to the GBS pho1-13 gene product can be identified in Streptococcus pyogenes, S. pneumoniae, S. salivarius, Escherichia coli, Yersinia enterocolitica, Aquifex aeolicus, Helicobacter pylori and Haemophilus influenzae. The S. pyogenes and S. pneumoniae homologues were identified from genome sequence data and no annotations were available as to the identity of the gene or gene products. In all other cases, the above homologues can be identified as ATP-dependent Clp protease proteolytic subunits. The catalytic activity of Clp proteases results in the hydrolysis of proteins to small peptides in the presence of ATP and magnesium (Giffard, P. M. et al. 1993. J. Gen. Microbiol. 139:913-920). Furthermore, the ClpP component of Clp proteases has been shown to be induced as part of the heat shock response (Kroh, H. E. and L. D. Simon. 1990. J. Bacteriol. 172:6026-6034) and it is probable that this subunit or the complete proteolytic domain would associated with the bacterial surface.
[0044]Immunisation studies, carried out as described above, yielded the following results.
TABLE-US-00001 No animals surviving at time (hrs) Treatment No animals 24 48 PBS 10 7 0 Pre-immunised 37 13 0 Immunised 38 17 9
EXAMPLE 2
[0045]A second clone was selected containing a plasmid designated pho1-14. This plasmid contained a gene (or part thereof), which complemented the leaderless phoA. The nucleotide and deduced amino acid sequences are shown as SEQ ID NOS. 3 and 4, respectively.
[0046]A comparison of the amino acid sequence of pho1-14 was performed.
[0047]Homologues to the GBS pho1-14 gene product can be identified in Streptococcus pyogenes, Enterococcus faecalis and Streptococcus pneumoniae. These homologues were identified from genome sequence data and no annotations were available as to the identity of the gene or gene products. Additionally, two possible homologues were also identified from Shigella flexneri (SpaR) and Yersinia pseudotuberculosis (YscT). These latter two homologues are related proteins, believed to be anchored in the bacterial membrane (Bergman, T. et al. 1994. J. Bacteriol. 176:2619-2626). In S. flexneri, the product of the spaR gene has been shown to be important for invasion of epithelial cells (Sasakawa, C. et al. 1993. J. Bacteriol. 175:2334-2346). Furthermore, the product of the spaR gene is also required for surface presentation of invasion plasmid antigens. The analogous protein in Y. pseudotuberculosis is a component of the Yop secretion system and is also important for virulence in this organism.
EXAMPLE 3
[0048]A third clone was selected containing a plasmid designated pho1-5. This plasmid contained a gene (or part thereof), which complemented the leaderless phoA. The nucleotide and deduced amino acid sequences are shown as SEQ ID NOS. 5 and 6.
[0049]A comparison of the amino acid sequence of pho1-5 was performed.
[0050]Homologues to the GBS pho1-5 gene product can only be identified in streptococcus pyogenes and Staphylococcus carnosus (sceA). The S. pyogenes homologue was identified from genome sequence data and no annotations were available as to the identity of the gene or gene products. Furthermore, little information is available on the function of the sceA gene product from S. carnosus. The sceA gene product shows some sequence similarity to the aggregation promoting protein from Lactobacillus gasseri. Based on analysis of the sceA gene product, this molecule contains a well-conserved signal sequence and is apparently secreted or associated with the bacterial cell surface.
EXAMPLE 4
[0051]A further clone was selected containing a plasmid designated pho3-3. This plasmid contained a gene (or part thereof), which complemented the leaderless phoA. The nucleotide and deduced amino acid sequences are shown as SEQ ID NOS. 7 and 8.
[0052]A comparison of the amino acid sequence of pho3-3 was performed.
[0053]Homologues to the GBS pho3-3 gene product can be identified in Streptococcus mutans (rmiC), (cpsM) S. pneumoniae and S. pyogenes. The S. pyogenes homologue was identified from genome sequence data and no annotations were available as to the identity of the gene or gene product. In S. pneumoniae, the homologue can be identified as dTDP-4-keto-6-deoxy glucose-3,5-epimerase. In the other two cases, the above homologues can be identified as dTDP-4-keto-L-rhamnose reductase (rmlC). In S. mutants, the gene encoding this enzyme, rmlC, is part of the rml locus. The rml locus consists of three genes which exhibit significant similarity to enzymes involved in the biosynthesis of dTDP-rhamnose, the immediate precursor of the rhamnose component in the S. mutans polysaccharide capsule (Tsukioka, Y. et al. 1997. J. Bacteriol. 179:1126-1134). An analogous locus has also been identified in S. pneumoniae (Coffey, T. J. et al. 1998. Mol. Micobiol. 17:73-83). Almost all Streptococci characteristically possess rhamnose in their cell wall associated polysaccharides (Schleifer, K. H. and R. Kilper-Balz. 1987. Syst. Appl. Microbiol. 10:1-19), and it is highly probable that dTDP-4-keto-L-rhamnose reductase would be associated with the outer surface in Streptococci.
EXAMPLE 5
[0054]A further clone was selected containing a plasmid designated pho2-10. This plasmid contained a gene (or part thereof), which complemented the leaderless phoA.
[0055]The nucleotide sequence is shown as SEQ ID NO. 9. From this, upstream and downstream coding regions were identified, and the deduced amino acid sequences shown as SEQ ID NOS. 10 and 11.
[0056]A comparison of the amino acid sequences of pho2-10 was performed.
[0057]Homologues to the GBS pho2-10 gene product can be identified in Streptococcus pyogenes, Enterococcus faecalis, Debaryomyces occidentalis (hatI) and Escherichia coli (trkD). The S. pyogenes and E. faecalis homologues were identified from genome sequence data and no annotations were available as to the identity of the gene or gene products. In the yeast D. occidentalis, the hak1 gene is a homologue of the trkD gene from E. coli (Banuelos, M. A. et al. 1995. EMBO J. 14:3021-3027). The trkd gene of E. coli is part of the kup potassium uptake system. The specific homolog identified here is the kup system potassium uptake protein. The kup system is a constitutive potassium uptake system in E. coli. The kup system potassium uptake protein contains a highly hydrophobic N-terminus that is predicted to span the membrane at least 12 times. Kup is not homologous to other known membrane protein sequences. There is no indication of ATP binding, and it is proposed that the system is driven by a chemiosmotic gradient (Schleyer, M. & E. P. Bakker, 1993. J. Bacteriol. 175:6925-6931).
EXAMPLE 6
[0058]A further clone was selected containing a plasmid designated pho2-15. 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. 12 and 13.
[0059]A comparison of the amino acid sequence of pho2-15 was performed.
[0060]Homologues to the GBS pho2-15 gene product can be identified in Streptococcus pyogenes, Streptococcus pneumoniae, Enterococcus faecalis and Escherichia coli (gatC and SgcC). The S. pyogenes, S. pneumoniae and E. faecalis homologues were identified from genome sequence data and no annotations were available as to the identity of the gene or gene products. In E. coli, the gatC and sgcC gene products can be identified as being the IIC component of phosphoenolypyruvate-dependent sugar phosphotransferase systems (PTS), a major carbohydrate active-transport system. In PTS systems, the IIC component is typically involved in binding of extracellular carbohydrates and forms a complex with the IID component to constitute a membrane channel (Nobelmann, B. and J. W. Lengeler. 1995. Biochim. Biophys. Acta 1262:69-72).
EXAMPLE 7
[0061]A further clone was selected containing a plasmid designated pho2-2. 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. 14 and 15, respectively.
[0062]A comparison of the amino acid sequence of pho2-2 was performed.
[0063]Homologues to the GBS pho2-2 gene product can be identified in Enterococcus faecalis, Escherichia coli (malK and afuC), Bacillus subtilis (glnO), Haemophilus influenzae (yebM and potA), Streptococcus pyogenes, Streptococcus pneumoniae and Salmonella typhimurium (malK). The E. faecalis, S. pyogenes and S. pneumoniae homologues were identified from genome sequence data and no annotations were available as to the identity of the gene or gene products. In all other cases, homologues represented ATP-binding transport proteins that are part of ABC type transporters. Many of the components of ABC type transporters are membrane or cell surface associated, as these systems are involved in the transport of macromolecules from the extracellular environment to the intracellular compartment.
EXAMPLE 8
[0064]A further clone was selected containing a plasmid designated pho3-14. 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. 16 and 17.
[0065]A comparison of the amino acid sequence of pho3-14 was performed and no homologues could be identified in any of the public databases. One homologue to the GBS pho3-14 gene product can be identified in Streptococcus pyogenes, but this homologue was identified from genome sequence data and no annotations were available as to the identity of the gene or gene product. Using this S. pyogenes homologue to search the public databases yielded no further information. Since the pho3-14 product complemented the leaderless phoA gene, it can be concluded that this protein (or part thereof) would most probably be located extracellularly.
EXAMPLE 9
[0066]A further clone was selected containing a plasmid designated pho3-17. 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. 18 and 19.
[0067]A comparison of the amino acid sequence of pho3-17 was performed.
[0068]Homologues to the GBS Pho3-17 gene product can be identified in Streptococcus mutans and Lactococcus lactis, with similarity being shown to N-acetyl muramidase. Similarity is also seen with an unidentified gene, yubE from Bacillus subtilis.
[0069]N-acetylmuramidase is an autolysin that is involved in cell division. Using this limited information along with the fact that pho3-17 complemented the leaderless phoA gene, it can be concluded that the pho3-17 product would most probably be located extracellularly.
EXAMPLE 10
[0070]A further clone was selected containing a plasmid designated pho3-18. 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. 20 and 21.
[0071]A comparison of the amino acid sequence of pho3-18 was performed.
[0072]Homologues to the GBS pho3-18 gene product can be identified in Streptococcus pyogenes and Streptococcus pneumoniae. These homologues were identified from genome sequence data and no annotations were available as to the identity of the gene or gene products. Using these S. pyogenes and S. pneumoniae homologues to search the public databases showed some similarity to outer surface and membrane spanning proteins. Since the ORF3-18 product complemented the leaderless phoA gene, it can be concluded that this protein (or part thereof) would most probably be located extracellularly.
EXAMPLE 11
[0073]A further clone was selected containing a plasmid designated pho3-1. 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. 22 and 23.
[0074]A comparison of the amino acid sequence of pho3-1 was performed.
[0075]Homologues to the GBS pho3-1 gene product can be identified in Streptococcus pyogenes, streptococcus pneumoniae, Bacillus subtilis (yutD) and Enterococcus faecalis. The S. pyogenes, S. pneumoniae and E. faecalis homologues were identified from genome sequence data and no annotations were available as to the identity of the gene or gene products. In B. subtilis, the function of the yutD gene product is unknown. It can be noted however, that the yutD gene is located on the B. subtilis chromosome in a region containing genes involved in cell wall synthesis. The fact that this DNA sequence complemented the leaderless phoA gene suggests that this gene product is extracellularly located.
EXAMPLE 12
[0076]A further clone was selected containing a plasmid designated pho3-21. 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. 24 and 25.
[0077]A comparison of the amino acid sequence of pho3-21 was performed.
[0078]Homologues to the GBS pho3-21 gene product can be identified in Streptococcus pyogenes, Streptococcus pneumoniae, Lactobacillus fermentum (bspA) and Lactobacillus reuteri (cnb). The S. pyogenes and S. pneumoniae homologues were identified from genome sequence data and no annotations were available as to the identity of the gene or gene products. In L. fermentum, the bspA gene product has been identified as being a basic cell surface-located protein that has some sequence similarity to family III of the bacterial solute-binding proteins (Turner, M. S. et al. 1997. J. Bacteriol. 179:3310-3316). In L. reuteri, the cnb gene product has been identified as a collagen binding protein that has some sequence similarity to the solute-binding component of bacterial ABC transporters (Roos, S. et al. 1996. FEMS Microbiol. Lett. 144:33-38).
EXAMPLE 13
[0079]A further clone was selected containing a plasmid designated pho3-22. 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. 26 and 27.
[0080]A comparison of the amino acid sequence of pho3-22 was performed.
[0081]Homologues to the GBS pho3-22 gene product can be identified in Enterococcus faecalis, Streptococcus equisimilis (lppC), Pseudomonas fluorescens (oprI) and Streptococcus thermophilus (orf142). The E. faecalis homolog was identified from genome sequence data and no annotations were available as to the identity of the gene or gene products. In S. equisimilis, the lppC gene product has been identified as being a lipoprotein that is homologous to the E(P4) outer membrane protein from Haemophilus influenzae (Gase, K. et al. 1997. Med. Microbiol. Immunol. 186:63-73). Likewise, the P. fluorescens oprI gene encodes a major outer membrane lipoprotein (Cornelis, P. et al. 1989. Mol. Microbiol. 3:421-428). In S. thermophilus, the orf142 product has been putatively identified as a cell surface exposed lipoprotein that may act as a receptor for the bacteriophages TP-J34 and Sfi21 (Neve, H. et al. 1998. Virology 241:61-72). The ORF3-22 product showed good similarity to the above homologues, particularly at the N-terminus. This is most likely the region required for complementation of the leaderless phoA gene, and therefore serves as a leader sequence.
EXAMPLE 14
[0082]A further clone was selected containing a plasmid designated pho3-23. This plasmid contained a gene (or part thereof), which complemented the leaderless phoA. The nucleotide and deduced amino acid sequences of the genes are shown as SEQ ID NOS. 28 and 29.
[0083]A comparison of the amino acid sequence of pho3-23 was performed.
[0084]Homologues to the GBS pho3-23 gene product can be identified in Streptococcus pyogenes, Streptococcus pneumoniae, Enterococcus faecalis and Streptococcus mutans (perM). The S. pyogenes, S. pneumoniae and E. faecalis homologues were identified from genome sequence data and no annotations were available as to the identity of the gene or gene products. In S. mutans, the perM gene product has been presumptively identified as a permease, but no other information is available as to the function of this protein. Considering that the pho3-23 coding region complements the leaderless phoA gene, it can be concluded that the pho3-17 gene product would most probably be located extracellularly.
EXAMPLE 15
[0085]A further clone was selected containing a plasmid designated pho3-24. 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. 30 and 31.
[0086]A comparison of the amino acid sequence of pho3-24 was performed.
[0087]Homologues to the GBS pho3-24 gene product can be identified in Streptococcus mutans (dltB), Streptococcus pneumoniae, Streptococcus pyogenes, Enterococcus faecalis, Lactobacillus casei (dltB) and Bacillus subtilis (dltB). The S. pneumoniae, S. pyogenes and E. faecalis homologues were identified from genome sequence data and no 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 transport of activated D-alanine through the cell membrane. The dltB gene product is involved in the biosynthesis of D-alanyl-lipoteichoic acid (Heaton, M. P. and F. C. Neuhaus. 1992. J. Bacteriol. 174:4707-4717). In L. casei and B. subtilis, 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
[0088]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.
[0089]A comparison of the amino acid sequence of pho3-29 was performed.
[0090]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. 61:5097-5105). The owp gene product from B. brevis is one of two major cell wall proteins involved in the surface layer lattice (Tsuboi, A. 1988. J. Bacteriol. 170:935-945). Finally, the oprI gene from P. aeruginosa encodes a major outer membrane lipoprotein precursor (Saint-Onge, A. et al. 1992. J. Gen. Microbiol. 138:733-741).
EXAMPLE 17
[0091]A further clone was selected containing a plasmid designated pho3-50. 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. 34 and 35.
[0092]A comparison of the amino acid sequence of pho3-50 was performed.
[0093]Homologues to the GBS pho3-50 gene product can be identified in a variety of Streptococci (penA, pbp2B, pbpB2), Borrelia burgdorferi (pbp2), Enterococcus faecalis(pbpC), Staphylococcus aureus (pbpA), Mycobacterium laprae(pbpB) and Helicobacter pylori (pbp2). In all cases, the above homologues can be identified as penicillin binding proteins (PBPs). Genes encoding penicillin binding proteins are often located in a cluster of genes associated with cell wall synthesis (Pucci, M. J. et al. 1997. J. Bacteriol. 179:5632-5635). Furthermore, PBPs are typically integrated into the cell wall of a bacterium with some or all of the protein being exposed on the outer bacterial surface.
Sequence CWU
1
351587DNAStreptococcus agalactiaeCDS(1)..(582) 1atg atc cca gta gta atc
gaa caa aca agt cgt ggt gaa cgt tct tat 48Met Ile Pro Val Val Ile
Glu Gln Thr Ser Arg Gly Glu Arg Ser Tyr1 5
10 15gat att tac tca cgt ctt tta aaa gat cgt att att
atg ttg aca ggc 96Asp Ile Tyr Ser Arg Leu Leu Lys Asp Arg Ile Ile
Met Leu Thr Gly 20 25 30caa
gtt gag gat aat atg gcc aat agt atc att gca cag tta ttg ttt 144Gln
Val Glu Asp Asn Met Ala Asn Ser Ile Ile Ala Gln Leu Leu Phe 35
40 45ctc gat gca caa gat aat aca aag gat
att tac ctt tat gtc aat aca 192Leu Asp Ala Gln Asp Asn Thr Lys Asp
Ile Tyr Leu Tyr Val Asn Thr 50 55
60cca ggt ggt tca gta tcg gct gga ctt gct att gtg gac acc atg aac
240Pro Gly Gly Ser Val Ser Ala Gly Leu Ala Ile Val Asp Thr Met Asn65
70 75 80ttc att aaa tcg gac
gta cag acg att gtt atg ggg atg gct gct tcg 288Phe Ile Lys Ser Asp
Val Gln Thr Ile Val Met Gly Met Ala Ala Ser 85
90 95atg gga acc att att gct tca agt ggt gct aaa
gga aaa cgt ttt atg 336Met Gly Thr Ile Ile Ala Ser Ser Gly Ala Lys
Gly Lys Arg Phe Met 100 105
110tta ccg aat gca gaa tat atg atc cac caa cca atg ggc gga aca ggc
384Leu Pro Asn Ala Glu Tyr Met Ile His Gln Pro Met Gly Gly Thr Gly
115 120 125gga ggt aca cag caa tct gat
atg gct atc gct gct gag cat ctt tta 432Gly Gly Thr Gln Gln Ser Asp
Met Ala Ile Ala Ala Glu His Leu Leu 130 135
140aaa acg cgt cat act tta gaa aaa atc tta gct gat aat tct ggt caa
480Lys Thr Arg His Thr Leu Glu Lys Ile Leu Ala Asp Asn Ser Gly Gln145
150 155 160tct att gaa aaa
gtc cat gat gat gca gag cgt gat cgt tgg atg agt 528Ser Ile Glu Lys
Val His Asp Asp Ala Glu Arg Asp Arg Trp Met Ser 165
170 175gct caa gaa aca ctt gat tat ggc ttt att
gat gaa atc atg gct aat 576Ala Gln Glu Thr Leu Asp Tyr Gly Phe Ile
Asp Glu Ile Met Ala Asn 180 185
190aat gaa taagg
587Asn Glu 2194PRTStreptococcus agalactiae 2Met Ile Pro Val Val Ile Glu
Gln Thr Ser Arg Gly Glu Arg Ser Tyr1 5 10
15Asp Ile Tyr Ser Arg Leu Leu Lys Asp Arg Ile Ile Met
Leu Thr Gly 20 25 30Gln Val
Glu Asp Asn Met Ala Asn Ser Ile Ile Ala Gln Leu Leu Phe 35
40 45Leu Asp Ala Gln Asp Asn Thr Lys Asp Ile
Tyr Leu Tyr Val Asn Thr 50 55 60Pro
Gly Gly Ser Val Ser Ala Gly Leu Ala Ile Val Asp Thr Met Asn65
70 75 80Phe Ile Lys Ser Asp Val
Gln Thr Ile Val Met Gly Met Ala Ala Ser 85
90 95Met Gly Thr Ile Ile Ala Ser Ser Gly Ala Lys Gly
Lys Arg Phe Met 100 105 110Leu
Pro Asn Ala Glu Tyr Met Ile His Gln Pro Met Gly Gly Thr Gly 115
120 125Gly Gly Thr Gln Gln Ser Asp Met Ala
Ile Ala Ala Glu His Leu Leu 130 135
140Lys Thr Arg His Thr Leu Glu Lys Ile Leu Ala Asp Asn Ser Gly Gln145
150 155 160Ser Ile Glu Lys
Val His Asp Asp Ala Glu Arg Asp Arg Trp Met Ser 165
170 175Ala Gln Glu Thr Leu Asp Tyr Gly Phe Ile
Asp Glu Ile Met Ala Asn 180 185
190Asn Glu3218DNAStreptococcus agalactiaeCDS(1)..(216) 3atc aga gca tat
tct ggt cct ctt tcg gtt ttc ctg cca cgt ttt aaa 48Ile Arg Ala Tyr
Ser Gly Pro Leu Ser Val Phe Leu Pro Arg Phe Lys1 5
10 15gct tgt gat ata ata gtc aat gtg agg agg
act atc atg tta ttt aag 96Ala Cys Asp Ile Ile Val Asn Val Arg Arg
Thr Ile Met Leu Phe Lys 20 25
30gaa aaa att cct gga cta ata tta tgc ttt att att gct ata cca tct
144Glu Lys Ile Pro Gly Leu Ile Leu Cys Phe Ile Ile Ala Ile Pro Ser
35 40 45tgg ttg ctt ggg ctt tat ctc cct
tta ata gga gca cca gtc ttt gct 192Trp Leu Leu Gly Leu Tyr Leu Pro
Leu Ile Gly Ala Pro Val Phe Ala 50 55
60atc ttg att gga ata att gtt gga tc
218Ile Leu Ile Gly Ile Ile Val Gly65
70472PRTStreptococcus agalactiae 4Ile Arg Ala Tyr Ser Gly Pro Leu Ser Val
Phe Leu Pro Arg Phe Lys1 5 10
15Ala Cys Asp Ile Ile Val Asn Val Arg Arg Thr Ile Met Leu Phe Lys
20 25 30Glu Lys Ile Pro Gly Leu
Ile Leu Cys Phe Ile Ile Ala Ile Pro Ser 35 40
45Trp Leu Leu Gly Leu Tyr Leu Pro Leu Ile Gly Ala Pro Val
Phe Ala 50 55 60Ile Leu Ile Gly Ile
Ile Val Gly65 705705DNAStreptococcus
agalactiaeCDS(1)..(705) 5atg aat aaa aga aga aaa tta tca aaa ttg aat gta
aaa aar caa cat 48Met Asn Lys Arg Arg Lys Leu Ser Lys Leu Asn Val
Lys Lys Gln His1 5 10
15tta gct tat gga gct atc act tta gta gcc ctt ttt tca tgt att ttg
96Leu Ala Tyr Gly Ala Ile Thr Leu Val Ala Leu Phe Ser Cys Ile Leu
20 25 30gct gta acg gtc atc ttt aaa
agt tca caa gtt act act gaa tct ttg 144Ala Val Thr Val Ile Phe Lys
Ser Ser Gln Val Thr Thr Glu Ser Leu 35 40
45tca aaa gca gat aaa gtt cgc gta gcc aaa aaa tca aaa atg act
aag 192Ser Lys Ala Asp Lys Val Arg Val Ala Lys Lys Ser Lys Met Thr
Lys 50 55 60gcg aca tct aaa tca aaa
gta gaa gat gta aaa cag gct cca aaa cct 240Ala Thr Ser Lys Ser Lys
Val Glu Asp Val Lys Gln Ala Pro Lys Pro65 70
75 80tct cag gca tct aat gaa gcc cca aaa tca agt
tct caa tct aca gaa 288Ser Gln Ala Ser Asn Glu Ala Pro Lys Ser Ser
Ser Gln Ser Thr Glu 85 90
95gct aat tct cag caa caa gtt act gcg agt gaa gag acg gct gta gaa
336Ala Asn Ser Gln Gln Gln Val Thr Ala Ser Glu Glu Thr Ala Val Glu
100 105 110caa gca gtt gta aca gaa
ata ccc ctg cta cca gtc agg cac aac aac 384Gln Ala Val Val Thr Glu
Ile Pro Leu Leu Pro Val Arg His Asn Asn 115 120
125ctt tat gct gtt act gag aca cct tac aac cct gct caa cca
cca gac 432Leu Tyr Ala Val Thr Glu Thr Pro Tyr Asn Pro Ala Gln Pro
Pro Asp 130 135 140caa gtg gcc agg tat
gag caa tgg aaa tac tgc cag gcg gtc gga tct 480Gln Val Ala Arg Tyr
Glu Gln Trp Lys Tyr Cys Gln Ala Val Gly Ser145 150
155 160gct gct gca gca caa atg gct gct gca aca
gga gtc cct cag tct act 528Ala Ala Ala Ala Gln Met Ala Ala Ala Thr
Gly Val Pro Gln Ser Thr 165 170
175tgg gaa cat att att gcc cgt gaa tca aat ggt aat cct aat gtt gct
576Trp Glu His Ile Ile Ala Arg Glu Ser Asn Gly Asn Pro Asn Val Ala
180 185 190aat gcc tca gga gct tca
gga ctt ttc caa acg atg cca ggt tgg ggt 624Asn Ala Ser Gly Ala Ser
Gly Leu Phe Gln Thr Met Pro Gly Trp Gly 195 200
205tca aca gct aca gtt cag gat caa gta att cag cta tta aag
ctt att 672Ser Thr Ala Thr Val Gln Asp Gln Val Ile Gln Leu Leu Lys
Leu Ile 210 215 220cgt gct caa ggg tta
tca gct ggg tac cag tga 705Arg Ala Gln Gly Leu
Ser Ala Gly Tyr Gln225 2306234PRTStreptococcus agalactiae
6Met Asn Lys Arg Arg Lys Leu Ser Lys Leu Asn Val Lys Lys Gln His1
5 10 15Leu Ala Tyr Gly Ala Ile
Thr Leu Val Ala Leu Phe Ser Cys Ile Leu 20 25
30Ala Val Thr Val Ile Phe Lys Ser Ser Gln Val Thr Thr
Glu Ser Leu 35 40 45Ser Lys Ala
Asp Lys Val Arg Val Ala Lys Lys Ser Lys Met Thr Lys 50
55 60Ala Thr Ser Lys Ser Lys Val Glu Asp Val Lys Gln
Ala Pro Lys Pro65 70 75
80Ser Gln Ala Ser Asn Glu Ala Pro Lys Ser Ser Ser Gln Ser Thr Glu
85 90 95Ala Asn Ser Gln Gln Gln
Val Thr Ala Ser Glu Glu Thr Ala Val Glu 100
105 110Gln Ala Val Val Thr Glu Ile Pro Leu Leu Pro Val
Arg His Asn Asn 115 120 125Leu Tyr
Ala Val Thr Glu Thr Pro Tyr Asn Pro Ala Gln Pro Pro Asp 130
135 140Gln Val Ala Arg Tyr Glu Gln Trp Lys Tyr Cys
Gln Ala Val Gly Ser145 150 155
160Ala Ala Ala Ala Gln Met Ala Ala Ala Thr Gly Val Pro Gln Ser Thr
165 170 175Trp Glu His Ile
Ile Ala Arg Glu Ser Asn Gly Asn Pro Asn Val Ala 180
185 190Asn Ala Ser Gly Ala Ser Gly Leu Phe Gln Thr
Met Pro Gly Trp Gly 195 200 205Ser
Thr Ala Thr Val Gln Asp Gln Val Ile Gln Leu Leu Lys Leu Ile 210
215 220Arg Ala Gln Gly Leu Ser Ala Gly Tyr
Gln225 2307594DNAStreptococcus agalactiaeCDS(1)..(594)
7atg act gaa cca ttt ttt gat aaa gaa tta act tgt cgc cca att gaa
48Met Thr Glu Pro Phe Phe Asp Lys Glu Leu Thr Cys Arg Pro Ile Glu1
5 10 15gcc att cct gaa ttg ttg
gaa ttc gat att acc gtt cgt gga gac aac 96Ala Ile Pro Glu Leu Leu
Glu Phe Asp Ile Thr Val Arg Gly Asp Asn 20 25
30cgt gga tgg ttc aaa gag aac ttt caa aaa gaa aaa atg
ata ccg ctt 144Arg Gly Trp Phe Lys Glu Asn Phe Gln Lys Glu Lys Met
Ile Pro Leu 35 40 45ggt ttc cca
gaa agc ttc ttt gag gca gac aaa cta caa aat aat att 192Gly Phe Pro
Glu Ser Phe Phe Glu Ala Asp Lys Leu Gln Asn Asn Ile 50
55 60tcg ttt aca aaa aaa aat act ttg cga ggt ctc cat
gca gag cct tgg 240Ser Phe Thr Lys Lys Asn Thr Leu Arg Gly Leu His
Ala Glu Pro Trp65 70 75
80gat aaa tat gtt tcg atc gct gat gaa gga cgt gtg atc ggt act tgg
288Asp Lys Tyr Val Ser Ile Ala Asp Glu Gly Arg Val Ile Gly Thr Trp
85 90 95gtt gac ctc cgt gaa ggt
gac agt ttt ggt aac gtt tac caa acg att 336Val Asp Leu Arg Glu Gly
Asp Ser Phe Gly Asn Val Tyr Gln Thr Ile 100
105 110atc gat gcc tca aaa ggt att ttt gtt cca cgc ggc
gtt gct aat ggt 384Ile Asp Ala Ser Lys Gly Ile Phe Val Pro Arg Gly
Val Ala Asn Gly 115 120 125ttc caa
gtt ctt tca gat aaa gca gct tat act tat ctc gtt aac gat 432Phe Gln
Val Leu Ser Asp Lys Ala Ala Tyr Thr Tyr Leu Val Asn Asp 130
135 140tat tgg gca ctt gaa ctc aaa cca aaa tat gct
ttc gtt aac tat gca 480Tyr Trp Ala Leu Glu Leu Lys Pro Lys Tyr Ala
Phe Val Asn Tyr Ala145 150 155
160gat cca aat cta ggc att cag tgg gaa aat ctw gaa gaa gca gaa gtc
528Asp Pro Asn Leu Gly Ile Gln Trp Glu Asn Leu Glu Glu Ala Glu Val
165 170 175tca gaa gca gat aag
aat cac cca ctt ctc aaa gat gta aaa cct ttg 576Ser Glu Ala Asp Lys
Asn His Pro Leu Leu Lys Asp Val Lys Pro Leu 180
185 190aag aag gaa gat ttg taa
594Lys Lys Glu Asp Leu 1958197PRTStreptococcus
agalactiae 8Met Thr Glu Pro Phe Phe Asp Lys Glu Leu Thr Cys Arg Pro Ile
Glu1 5 10 15Ala Ile Pro
Glu Leu Leu Glu Phe Asp Ile Thr Val Arg Gly Asp Asn 20
25 30Arg Gly Trp Phe Lys Glu Asn Phe Gln Lys
Glu Lys Met Ile Pro Leu 35 40
45Gly Phe Pro Glu Ser Phe Phe Glu Ala Asp Lys Leu Gln Asn Asn Ile 50
55 60Ser Phe Thr Lys Lys Asn Thr Leu Arg
Gly Leu His Ala Glu Pro Trp65 70 75
80Asp Lys Tyr Val Ser Ile Ala Asp Glu Gly Arg Val Ile Gly
Thr Trp 85 90 95Val Asp
Leu Arg Glu Gly Asp Ser Phe Gly Asn Val Tyr Gln Thr Ile 100
105 110Ile Asp Ala Ser Lys Gly Ile Phe Val
Pro Arg Gly Val Ala Asn Gly 115 120
125Phe Gln Val Leu Ser Asp Lys Ala Ala Tyr Thr Tyr Leu Val Asn Asp
130 135 140Tyr Trp Ala Leu Glu Leu Lys
Pro Lys Tyr Ala Phe Val Asn Tyr Ala145 150
155 160Asp Pro Asn Leu Gly Ile Gln Trp Glu Asn Leu Glu
Glu Ala Glu Val 165 170
175Ser Glu Ala Asp Lys Asn His Pro Leu Leu Lys Asp Val Lys Pro Leu
180 185 190Lys Lys Glu Asp Leu
19591217DNAStreptococcus agalactiaemisc_feature(571)..(571)N can be A, G,
C, or T. 9tat tat tta atc gga ggg ttg gca gaa atg caa cat gtc aat cat tct
48Tyr Tyr Leu Ile Gly Gly Leu Ala Glu Met Gln His Val Asn His Ser1
5 10 15tct ttt gat aaa
gca tca aaa gca gga ttt att att gct tta ggc att 96Ser Phe Asp Lys
Ala Ser Lys Ala Gly Phe Ile Ile Ala Leu Gly Ile 20
25 30gtt tat gga gat att ggt aca agc cca ctc tat
acg atg caa tca ttg 144Val Tyr Gly Asp Ile Gly Thr Ser Pro Leu Tyr
Thr Met Gln Ser Leu 35 40 45gtt
gaa aac caa ggt ggt att tct agt gtc aca gaa tcg ttt atc tta 192Val
Glu Asn Gln Gly Gly Ile Ser Ser Val Thr Glu Ser Phe Ile Leu 50
55 60ggt tct ata tct tta atc ata tgg acc ttg
aca ctt att aca act atc 240Gly Ser Ile Ser Leu Ile Ile Trp Thr Leu
Thr Leu Ile Thr Thr Ile65 70 75
80aag tat gtg ctt gta gct tta aag gcg gat aat cac cac gaa ggt
ggt 288Lys Tyr Val Leu Val Ala Leu Lys Ala Asp Asn His His Glu Gly
Gly 85 90 95att ttt tct
tta tat acc ctt gtt aga aaa atg aca cct tgg tta att 336Ile Phe Ser
Leu Tyr Thr Leu Val Arg Lys Met Thr Pro Trp Leu Ile 100
105 110gtt ccg gct gtt att gga ggt gca acc ttg
ttg tca gat gga gct ttg 384Val Pro Ala Val Ile Gly Gly Ala Thr Leu
Leu Ser Asp Gly Ala Leu 115 120
125acg cca gct gta acc gta ctt cag ccg tta agg att aaa gta gtt cct
432Thr Pro Ala Val Thr Val Leu Gln Pro Leu Arg Ile Lys Val Val Pro 130
135 140agt ttg cag cat att tcc aga atc
aga gta tgt tat ttt gcg acc ttg 480Ser Leu Gln His Ile Ser Arg Ile
Arg Val Cys Tyr Phe Ala Thr Leu145 150
155 160tta ttt act gtt act ttt gcc atc caa ggt ttg gaa
cgg gtg tta ttg 528Leu Phe Thr Val Thr Phe Ala Ile Gln Gly Leu Glu
Arg Val Leu Leu 165 170
175gaa tta ttg gcc att atg tta tat ggt ttg cct ttt ggt tta
570Glu Leu Leu Ala Ile Met Leu Tyr Gly Leu Pro Phe Gly Leu 180
185 190ncggtctcct tatagttttg cccatccaga
agttttcaag cattaatcca tactacggtt 630tgaaattgtt atttagtcca gagaatcata
aaggtatttt tattttag gat cta ttt 687
Asp Leu Phetcc tgg cga caa acg gga gca gaa gca cta tac
tct gac tta ggt cat 735Ser Trp Arg Gln Thr Gly Ala Glu Ala Leu Tyr
Ser Asp Leu Gly His 195 200 205gtt ggg
cgt gga aat ata cat gtt tca tgg ccg ttc gtt aag gtt gcc 783Val Gly
Arg Gly Asn Ile His Val Ser Trp Pro Phe Val Lys Val Ala210
215 220 225att ata ctt tct tat tgt ggg
caa ggg gca tgg att tta gct aat aag 831Ile Ile Leu Ser Tyr Cys Gly
Gln Gly Ala Trp Ile Leu Ala Asn Lys 230
235 240aac gca gga aat gaa ttg aat ccc ttt ttt gct agt
att cct tcg caa 879Asn Ala Gly Asn Glu Leu Asn Pro Phe Phe Ala Ser
Ile Pro Ser Gln 245 250 255ttt
aca atg cat gtc gtt att tta gct act ttg gca gct atc atc gct 927Phe
Thr Met His Val Val Ile Leu Ala Thr Leu Ala Ala Ile Ile Ala 260
265 270tca cag gca ctg att tct ggatcaattt
accttaagtt ctgagctatg 975Ser Gln Ala Leu Ile Ser
275cgactaaaaa tattcccaca atttcgttca acttatcctg ttgacaatat tgggtcaaac
1035ctacatacct ggtattaatt ggttcttatt tgccattaca acctctattg gtttgctttt
1095taagacttca gcgcacatgg aagcagcata tggattagcg ataacaatta cgatgctaat
1155gacaactatt ttactgtctt tctttttaat tcaaaaagga gtaaagagag gttttagcta
1215tt
121710190PRTStreptococcus agalactiae 10Tyr Tyr Leu Ile Gly Gly Leu Ala
Glu Met Gln His Val Asn His Ser1 5 10
15Ser Phe Asp Lys Ala Ser Lys Ala Gly Phe Ile Ile Ala Leu
Gly Ile 20 25 30Val Tyr Gly
Asp Ile Gly Thr Ser Pro Leu Tyr Thr Met Gln Ser Leu 35
40 45Val Glu Asn Gln Gly Gly Ile Ser Ser Val Thr
Glu Ser Phe Ile Leu 50 55 60Gly Ser
Ile Ser Leu Ile Ile Trp Thr Leu Thr Leu Ile Thr Thr Ile65
70 75 80Lys Tyr Val Leu Val Ala Leu
Lys Ala Asp Asn His His Glu Gly Gly 85 90
95Ile Phe Ser Leu Tyr Thr Leu Val Arg Lys Met Thr Pro
Trp Leu Ile 100 105 110Val Pro
Ala Val Ile Gly Gly Ala Thr Leu Leu Ser Asp Gly Ala Leu 115
120 125Thr Pro Ala Val Thr Val Leu Gln Pro Leu
Arg Ile Lys Val Val Pro 130 135 140Ser
Leu Gln His Ile Ser Arg Ile Arg Val Cys Tyr Phe Ala Thr Leu145
150 155 160Leu Phe Thr Val Thr Phe
Ala Ile Gln Gly Leu Glu Arg Val Leu Leu 165
170 175Glu Leu Leu Ala Ile Met Leu Tyr Gly Leu Pro Phe
Gly Leu 180 185
1901189PRTStreptococcus agalactiae 11Asp Leu Phe Ser Trp Arg Gln Thr Gly
Ala Glu Ala Leu Tyr Ser Asp1 5 10
15Leu Gly His Val Gly Arg Gly Asn Ile His Val Ser Trp Pro Phe
Val 20 25 30Lys Val Ala Ile
Ile Leu Ser Tyr Cys Gly Gln Gly Ala Trp Ile Leu 35
40 45Ala Asn Lys Asn Ala Gly Asn Glu Leu Asn Pro Phe
Phe Ala Ser Ile 50 55 60Pro Ser Gln
Phe Thr Met His Val Val Ile Leu Ala Thr Leu Ala Ala65 70
75 80Ile Ile Ala Ser Gln Ala Leu Ile
Ser 8512378DNAStreptococcus agalactiaeCDS(1)..(378) 12atg
cag gta ttt tta aat att gtc aat aaa ttc ttt gat cca gtt att 48Met
Gln Val Phe Leu Asn Ile Val Asn Lys Phe Phe Asp Pro Val Ile1
5 10 15cat atg ggt tcg gga gtt gtg
atg cta att gtc atg aca ggt tta gcc 96His Met Gly Ser Gly Val Val
Met Leu Ile Val Met Thr Gly Leu Ala 20 25
30atg ata ttt gga gtg aag ttt tct aaa gca ctt gaa ggt ggt
att aag 144Met Ile Phe Gly Val Lys Phe Ser Lys Ala Leu Glu Gly Gly
Ile Lys 35 40 45tta gct att gct
ctt acg ggt att ggt gct att att ggt att tta act 192Leu Ala Ile Ala
Leu Thr Gly Ile Gly Ala Ile Ile Gly Ile Leu Thr 50 55
60ggt gct ttt tcc gaa tca ctt caa gct ttt gtt aaa aat
aca gga atc 240Gly Ala Phe Ser Glu Ser Leu Gln Ala Phe Val Lys Asn
Thr Gly Ile65 70 75
80aat cta agc att att gac gtt ggt tgg gct cca tta gca act att aca
288Asn Leu Ser Ile Ile Asp Val Gly Trp Ala Pro Leu Ala Thr Ile Thr
85 90 95tgg gga tca cca tat acg
ctt tac ttc tta tta atc atg ctt att gtc 336Trp Gly Ser Pro Tyr Thr
Leu Tyr Phe Leu Leu Ile Met Leu Ile Val 100
105 110aat att gtt atg att gtt atg aaa aaa aaa cgg ata
cct tag 378Asn Ile Val Met Ile Val Met Lys Lys Lys Arg Ile
Pro 115 120
12513125PRTStreptococcus agalactiae 13Met Gln Val Phe Leu Asn Ile Val Asn
Lys Phe Phe Asp Pro Val Ile1 5 10
15His Met Gly Ser Gly Val Val Met Leu Ile Val Met Thr Gly Leu
Ala 20 25 30Met Ile Phe Gly
Val Lys Phe Ser Lys Ala Leu Glu Gly Gly Ile Lys 35
40 45Leu Ala Ile Ala Leu Thr Gly Ile Gly Ala Ile Ile
Gly Ile Leu Thr 50 55 60Gly Ala Phe
Ser Glu Ser Leu Gln Ala Phe Val Lys Asn Thr Gly Ile65 70
75 80Asn Leu Ser Ile Ile Asp Val Gly
Trp Ala Pro Leu Ala Thr Ile Thr 85 90
95Trp Gly Ser Pro Tyr Thr Leu Tyr Phe Leu Leu Ile Met Leu
Ile Val 100 105 110Asn Ile Val
Met Ile Val Met Lys Lys Lys Arg Ile Pro 115 120
12514705DNAStreptococcus agalactiaeCDS(118)..(705)
14ggatcgggcg caagcttaac gattcttttt aaaatcatta aattttaaaa caaatttcag
60acatattgcc aaagttttga tattattact ataatatagt ttgtagagga gaataat
117atg ggc caa gaa cct atc atc gaa tat caa aat atc aat aaa gtg tat
165Met Gly Gln Glu Pro Ile Ile Glu Tyr Gln Asn Ile Asn Lys Val Tyr1
5 10 15ggg gaa aat gtt gcg gtt
gaa gat att aac ctt aaa att tac cct ggt 213Gly Glu Asn Val Ala Val
Glu Asp Ile Asn Leu Lys Ile Tyr Pro Gly 20 25
30gat ttc gtt tgt ttc atc ggt acg agt gga tca ggt aaa
aca aca tta 261Asp Phe Val Cys Phe Ile Gly Thr Ser Gly Ser Gly Lys
Thr Thr Leu 35 40 45atg cgt atg
gtt aac cat atg tta aaa cca aca aat ggt act cta tta 309Met Arg Met
Val Asn His Met Leu Lys Pro Thr Asn Gly Thr Leu Leu 50
55 60ttt aag gga aaa gat atc tct act att aac ccc att
gaa tta aga cgc 357Phe Lys Gly Lys Asp Ile Ser Thr Ile Asn Pro Ile
Glu Leu Arg Arg65 70 75
80aga att gga tat gtt atc caa aac att ggt tta atg cct cat atg acc
405Arg Ile Gly Tyr Val Ile Gln Asn Ile Gly Leu Met Pro His Met Thr
85 90 95att tac gaa aat ata gtt
ctt gta cca aaa tta ttg aaa tgg tca gaa 453Ile Tyr Glu Asn Ile Val
Leu Val Pro Lys Leu Leu Lys Trp Ser Glu 100
105 110gaa gct aaa aga gct aaa gca agg gaa ctt att aaa
tta gtt gaa tta 501Glu Ala Lys Arg Ala Lys Ala Arg Glu Leu Ile Lys
Leu Val Glu Leu 115 120 125ccc gaa
gaa tat ttg gat cgc tac cct agt gag ttg tct ggc ggt cag 549Pro Glu
Glu Tyr Leu Asp Arg Tyr Pro Ser Glu Leu Ser Gly Gly Gln 130
135 140caa caa cgt atc ggt gtc att cgc gct ctt gca
gca gac caa gat att 597Gln Gln Arg Ile Gly Val Ile Arg Ala Leu Ala
Ala Asp Gln Asp Ile145 150 155
160att tta atg gat gag cct ttt gga gct ctg gat cct att act aga gaa
645Ile Leu Met Asp Glu Pro Phe Gly Ala Leu Asp Pro Ile Thr Arg Glu
165 170 175ggt att caa gac ttt
agt caa gtc tct tca gga aga aat ggg gga aaa 693Gly Ile Gln Asp Phe
Ser Gln Val Ser Ser Gly Arg Asn Gly Gly Lys 180
185 190cta tca tct tag
705Leu Ser Ser 19515195PRTStreptococcus
agalactiae 15Met Gly Gln Glu Pro Ile Ile Glu Tyr Gln Asn Ile Asn Lys Val
Tyr1 5 10 15Gly Glu Asn
Val Ala Val Glu Asp Ile Asn Leu Lys Ile Tyr Pro Gly 20
25 30Asp Phe Val Cys Phe Ile Gly Thr Ser Gly
Ser Gly Lys Thr Thr Leu 35 40
45Met Arg Met Val Asn His Met Leu Lys Pro Thr Asn Gly Thr Leu Leu 50
55 60Phe Lys Gly Lys Asp Ile Ser Thr Ile
Asn Pro Ile Glu Leu Arg Arg65 70 75
80Arg Ile Gly Tyr Val Ile Gln Asn Ile Gly Leu Met Pro His
Met Thr 85 90 95Ile Tyr
Glu Asn Ile Val Leu Val Pro Lys Leu Leu Lys Trp Ser Glu 100
105 110Glu Ala Lys Arg Ala Lys Ala Arg Glu
Leu Ile Lys Leu Val Glu Leu 115 120
125Pro Glu Glu Tyr Leu Asp Arg Tyr Pro Ser Glu Leu Ser Gly Gly Gln
130 135 140Gln Gln Arg Ile Gly Val Ile
Arg Ala Leu Ala Ala Asp Gln Asp Ile145 150
155 160Ile Leu Met Asp Glu Pro Phe Gly Ala Leu Asp Pro
Ile Thr Arg Glu 165 170
175Gly Ile Gln Asp Phe Ser Gln Val Ser Ser Gly Arg Asn Gly Gly Lys
180 185 190Leu Ser Ser
19516367DNAStreptococcus agalactiaeCDS(1)..(366) 16atc cct tat agt gat
gtt ttt gct aca gga gga ttt tta tac tat gta 48Ile Pro Tyr Ser Asp
Val Phe Ala Thr Gly Gly Phe Leu Tyr Tyr Val1 5
10 15acg att gct cta agt tac ctt tta ggg tct agt
atc tgg tta ttt att 96Thr Ile Ala Leu Ser Tyr Leu Leu Gly Ser Ser
Ile Trp Leu Phe Ile 20 25
30gta cag ttt att gct tac tat gta tct gga att tat ttt tat aaa tta
144Val Gln Phe Ile Ala Tyr Tyr Val Ser Gly Ile Tyr Phe Tyr Lys Leu
35 40 45gtt tat tat gtg gca caa agt gaa
att gtc tcg ata ggc atg acg ttg 192Val Tyr Tyr Val Ala Gln Ser Glu
Ile Val Ser Ile Gly Met Thr Leu 50 55
60att ttc tat ata atg aat att gtc tta gga ttc ggt ggt atg tac cca
240Ile Phe Tyr Ile Met Asn Ile Val Leu Gly Phe Gly Gly Met Tyr Pro65
70 75 80ata cag tgg gca tta
cct ttt atg ctc att tcg cta tgg ttt tta att 288Ile Gln Trp Ala Leu
Pro Phe Met Leu Ile Ser Leu Trp Phe Leu Ile 85
90 95aaa ttt tgt gtc gat aat atc gtt gat gaa gca
ttt ata ttt tat ggt 336Lys Phe Cys Val Asp Asn Ile Val Asp Glu Ala
Phe Ile Phe Tyr Gly 100 105
110att tta gca gca ttc tca cta ttt ata gat c
367Ile Leu Ala Ala Phe Ser Leu Phe Ile Asp 115
12017122PRTStreptococcus agalactiae 17Ile Pro Tyr Ser Asp Val Phe Ala Thr
Gly Gly Phe Leu Tyr Tyr Val1 5 10
15Thr Ile Ala Leu Ser Tyr Leu Leu Gly Ser Ser Ile Trp Leu Phe
Ile 20 25 30Val Gln Phe Ile
Ala Tyr Tyr Val Ser Gly Ile Tyr Phe Tyr Lys Leu 35
40 45Val Tyr Tyr Val Ala Gln Ser Glu Ile Val Ser Ile
Gly Met Thr Leu 50 55 60Ile Phe Tyr
Ile Met Asn Ile Val Leu Gly Phe Gly Gly Met Tyr Pro65 70
75 80Ile Gln Trp Ala Leu Pro Phe Met
Leu Ile Ser Leu Trp Phe Leu Ile 85 90
95Lys Phe Cys Val Asp Asn Ile Val Asp Glu Ala Phe Ile Phe
Tyr Gly 100 105 110Ile Leu Ala
Ala Phe Ser Leu Phe Ile Asp 115
12018570DNAStreptococcus agalactiaeCDS(1)..(570) 18atg agg aaa cgt ttt
tcc ttg cta aat ttt att gtt gtt act ttt att 48Met Arg Lys Arg Phe
Ser Leu Leu Asn Phe Ile Val Val Thr Phe Ile1 5
10 15ttc ttt ttc ttt att ctt ttt ccg ctt tta aac
cat aag gga aaa gta 96Phe Phe Phe Phe Ile Leu Phe Pro Leu Leu Asn
His Lys Gly Lys Val 20 25
30gat gct aat tct agg cag agt gtt acc tac acc aaa gaa gaa ttt ata
144Asp Ala Asn Ser Arg Gln Ser Val Thr Tyr Thr Lys Glu Glu Phe Ile
35 40 45caa aaa att gtg cca gat gcg caa
gat cta gga aag tcg tac ggt att 192Gln Lys Ile Val Pro Asp Ala Gln
Asp Leu Gly Lys Ser Tyr Gly Ile 50 55
60cgt cct tca ttt att att gca cag gcg gct ttg gat tct gat ttc gga
240Arg Pro Ser Phe Ile Ile Ala Gln Ala Ala Leu Asp Ser Asp Phe Gly65
70 75 80gag aaa tat agc tat
agt atc ata atc tgt tgg ttg ctt gca gaa cca 288Glu Lys Tyr Ser Tyr
Ser Ile Ile Ile Cys Trp Leu Leu Ala Glu Pro 85
90 95gga acg ccc tca att acc tta aat gat agt agt
aca gga aaa aaa cag 336Gly Thr Pro Ser Ile Thr Leu Asn Asp Ser Ser
Thr Gly Lys Lys Gln 100 105
110gaa aag caa ttt act cat tat aaa tct tgg aag tat tca atg gat gat
384Glu Lys Gln Phe Thr His Tyr Lys Ser Trp Lys Tyr Ser Met Asp Asp
115 120 125tac ctt gct cat ata aaa tct
gga gcg aca ggc aaa aaa gat tca tat 432Tyr Leu Ala His Ile Lys Ser
Gly Ala Thr Gly Lys Lys Asp Ser Tyr 130 135
140act ata atg gtg tct gtt aaa aat cca aaa act tta gtg caa aaa tta
480Thr Ile Met Val Ser Val Lys Asn Pro Lys Thr Leu Val Gln Lys Leu145
150 155 160caa gat agt ggt
ttt gat aat gac aaa aag tac gct aaa aaa atg acg 528Gln Asp Ser Gly
Phe Asp Asn Asp Lys Lys Tyr Ala Lys Lys Met Thr 165
170 175gaa atc att gat ttg tat gat tta aca aga
tat gat aag tga 570Glu Ile Ile Asp Leu Tyr Asp Leu Thr Arg
Tyr Asp Lys 180 18519189PRTStreptococcus
agalactiae 19Met Arg Lys Arg Phe Ser Leu Leu Asn Phe Ile Val Val Thr Phe
Ile1 5 10 15Phe Phe Phe
Phe Ile Leu Phe Pro Leu Leu Asn His Lys Gly Lys Val 20
25 30Asp Ala Asn Ser Arg Gln Ser Val Thr Tyr
Thr Lys Glu Glu Phe Ile 35 40
45Gln Lys Ile Val Pro Asp Ala Gln Asp Leu Gly Lys Ser Tyr Gly Ile 50
55 60Arg Pro Ser Phe Ile Ile Ala Gln Ala
Ala Leu Asp Ser Asp Phe Gly65 70 75
80Glu Lys Tyr Ser Tyr Ser Ile Ile Ile Cys Trp Leu Leu Ala
Glu Pro 85 90 95Gly Thr
Pro Ser Ile Thr Leu Asn Asp Ser Ser Thr Gly Lys Lys Gln 100
105 110Glu Lys Gln Phe Thr His Tyr Lys Ser
Trp Lys Tyr Ser Met Asp Asp 115 120
125Tyr Leu Ala His Ile Lys Ser Gly Ala Thr Gly Lys Lys Asp Ser Tyr
130 135 140Thr Ile Met Val Ser Val Lys
Asn Pro Lys Thr Leu Val Gln Lys Leu145 150
155 160Gln Asp Ser Gly Phe Asp Asn Asp Lys Lys Tyr Ala
Lys Lys Met Thr 165 170
175Glu Ile Ile Asp Leu Tyr Asp Leu Thr Arg Tyr Asp Lys 180
18520978DNAStreptococcus agalactiaeCDS(1)..(978) 20atg ctt
gtc atc att ttg atc att gta cta gct agt ctg aca gtg acg 48Met Leu
Val Ile Ile Leu Ile Ile Val Leu Ala Ser Leu Thr Val Thr1 5
10 15ata att tct tac cca aaa atg acg
gaa tta aca aag tcc gtt gaa aaa 96Ile Ile Ser Tyr Pro Lys Met Thr
Glu Leu Thr Lys Ser Val Glu Lys 20 25
30caa ctt gaa gat aat gct gat aat cta tca gac caa ctg aca tat
cag 144Gln Leu Glu Asp Asn Ala Asp Asn Leu Ser Asp Gln Leu Thr Tyr
Gln 35 40 45ata gaa gtg gcg caa
aaa gat caa atc tac gtg act aat cag cta aac 192Ile Glu Val Ala Gln
Lys Asp Gln Ile Tyr Val Thr Asn Gln Leu Asn 50 55
60cgt atg caa cag gaa att atc agt cgc tta ccg ata tgc gta
cag aat 240Arg Met Gln Gln Glu Ile Ile Ser Arg Leu Pro Ile Cys Val
Gln Asn65 70 75 80aaa
tca gca tta acg gag agt cga gat cga tca gac aaa cgc ttg gaa 288Lys
Ser Ala Leu Thr Glu Ser Arg Asp Arg Ser Asp Lys Arg Leu Glu
85 90 95ttg att aac tcc aat tta tct
cag tca gtt cag aaa atg caa gat tca 336Leu Ile Asn Ser Asn Leu Ser
Gln Ser Val Gln Lys Met Gln Asp Ser 100 105
110atg aaa aac gct tgg atc aaa tgc gcc aaa ctg ttg agg aaa
agc tgg 384Met Lys Asn Ala Trp Ile Lys Cys Ala Lys Leu Leu Arg Lys
Ser Trp 115 120 125aaa aaa cgc tac
aaa cgc gtt gca aac ttc ttt gaa act gta tcg cgt 432Lys Lys Arg Tyr
Lys Arg Val Ala Asn Phe Phe Glu Thr Val Ser Arg 130
135 140caa cta gag agc gtc aat caa ggt ctg ggt aga tgg
aaa ctg tgc caa 480Gln Leu Glu Ser Val Asn Gln Gly Leu Gly Arg Trp
Lys Leu Cys Gln145 150 155
160gat gtt ggt acc act gaa caa agt ctg tca aat act aag aca agg gga
528Asp Val Gly Thr Thr Glu Gln Ser Leu Ser Asn Thr Lys Thr Arg Gly
165 170 175ata tta ggg gag tta
caa ctc ggt caa att ata gaa gat att atg aca 576Ile Leu Gly Glu Leu
Gln Leu Gly Gln Ile Ile Glu Asp Ile Met Thr 180
185 190gtt agt caa tat gag aga gaa ttt cct acg gtg tct
ggc tct tct gag 624Val Ser Gln Tyr Glu Arg Glu Phe Pro Thr Val Ser
Gly Ser Ser Glu 195 200 205cgt gtt
gaa tat gct att aaa tac ctg gaa atg gtc agg gag att ata 672Arg Val
Glu Tyr Ala Ile Lys Tyr Leu Glu Met Val Arg Glu Ile Ile 210
215 220tct att tgc cta ttg act cta agt ttc tct aga
aga tta tta ccg att 720Ser Ile Cys Leu Leu Thr Leu Ser Phe Ser Arg
Arg Leu Leu Pro Ile225 230 235
240ggg aga tgc tta tgg aat tgg gtg acc agg ttc aaa tgg aac tct att
768Gly Arg Cys Leu Trp Asn Trp Val Thr Arg Phe Lys Trp Asn Ser Ile
245 250 255cgt aat ctt tac tgg
gca agt att cgt aaa ttt gca aaa gat ata aac 816Arg Asn Leu Tyr Trp
Ala Ser Ile Arg Lys Phe Ala Lys Asp Ile Asn 260
265 270aat aag tac tta aat cct cct gaa acg aca aat ttt
ggt atc atg ttc 864Asn Lys Tyr Leu Asn Pro Pro Glu Thr Thr Asn Phe
Gly Ile Met Phe 275 280 285tta cca
act gaa ggg ctc tat tct gaa gtg gta aga aat gca aca ttc 912Leu Pro
Thr Glu Gly Leu Tyr Ser Glu Val Val Arg Asn Ala Thr Phe 290
295 300ttt gat agt cta aga cgt gac gaa aat att gta
gta gct gga ccg tca 960Phe Asp Ser Leu Arg Arg Asp Glu Asn Ile Val
Val Ala Gly Pro Ser305 310 315
320acc tta tct gct tac taa
978Thr Leu Ser Ala Tyr 32521325PRTStreptococcus
agalactiae 21Met Leu Val Ile Ile Leu Ile Ile Val Leu Ala Ser Leu Thr Val
Thr1 5 10 15Ile Ile Ser
Tyr Pro Lys Met Thr Glu Leu Thr Lys Ser Val Glu Lys 20
25 30Gln Leu Glu Asp Asn Ala Asp Asn Leu Ser
Asp Gln Leu Thr Tyr Gln 35 40
45Ile Glu Val Ala Gln Lys Asp Gln Ile Tyr Val Thr Asn Gln Leu Asn 50
55 60Arg Met Gln Gln Glu Ile Ile Ser Arg
Leu Pro Ile Cys Val Gln Asn65 70 75
80Lys Ser Ala Leu Thr Glu Ser Arg Asp Arg Ser Asp Lys Arg
Leu Glu 85 90 95Leu Ile
Asn Ser Asn Leu Ser Gln Ser Val Gln Lys Met Gln Asp Ser 100
105 110Met Lys Asn Ala Trp Ile Lys Cys Ala
Lys Leu Leu Arg Lys Ser Trp 115 120
125Lys Lys Arg Tyr Lys Arg Val Ala Asn Phe Phe Glu Thr Val Ser Arg
130 135 140Gln Leu Glu Ser Val Asn Gln
Gly Leu Gly Arg Trp Lys Leu Cys Gln145 150
155 160Asp Val Gly Thr Thr Glu Gln Ser Leu Ser Asn Thr
Lys Thr Arg Gly 165 170
175Ile Leu Gly Glu Leu Gln Leu Gly Gln Ile Ile Glu Asp Ile Met Thr
180 185 190Val Ser Gln Tyr Glu Arg
Glu Phe Pro Thr Val Ser Gly Ser Ser Glu 195 200
205Arg Val Glu Tyr Ala Ile Lys Tyr Leu Glu Met Val Arg Glu
Ile Ile 210 215 220Ser Ile Cys Leu Leu
Thr Leu Ser Phe Ser Arg Arg Leu Leu Pro Ile225 230
235 240Gly Arg Cys Leu Trp Asn Trp Val Thr Arg
Phe Lys Trp Asn Ser Ile 245 250
255Arg Asn Leu Tyr Trp Ala Ser Ile Arg Lys Phe Ala Lys Asp Ile Asn
260 265 270Asn Lys Tyr Leu Asn
Pro Pro Glu Thr Thr Asn Phe Gly Ile Met Phe 275
280 285Leu Pro Thr Glu Gly Leu Tyr Ser Glu Val Val Arg
Asn Ala Thr Phe 290 295 300Phe Asp Ser
Leu Arg Arg Asp Glu Asn Ile Val Val Ala Gly Pro Ser305
310 315 320Thr Leu Ser Ala Tyr
32522579DNAStreptococcus agalactiaeCDS(1)..(579) 22atg cga aaa gaa
gtg aca cca gag atg ctt aac tat aat aag tat cct 48Met Arg Lys Glu
Val Thr Pro Glu Met Leu Asn Tyr Asn Lys Tyr Pro1 5
10 15ggc cca cag ttt att cac ttt gaa aat atc
gtt aaa agt gat gat att 96Gly Pro Gln Phe Ile His Phe Glu Asn Ile
Val Lys Ser Asp Asp Ile 20 25
30gaa ttt caa ctt gtt att aat gaa aaa tca gct ttt gat gtg act gtc
144Glu Phe Gln Leu Val Ile Asn Glu Lys Ser Ala Phe Asp Val Thr Val
35 40 45ttt gga caa cgt ttt tct gag att
tta tta aaa tat gat ttt atc gtt 192Phe Gly Gln Arg Phe Ser Glu Ile
Leu Leu Lys Tyr Asp Phe Ile Val 50 55
60ggc gat tgg ggt aac gag cag ttg agg cta aga ggc ttt tac aaa gat
240Gly Asp Trp Gly Asn Glu Gln Leu Arg Leu Arg Gly Phe Tyr Lys Asp65
70 75 80gct agt acg att aga
aaa aat agc cgg att tca cgt tta gaa gat tat 288Ala Ser Thr Ile Arg
Lys Asn Ser Arg Ile Ser Arg Leu Glu Asp Tyr 85
90 95att aaa gag tat tgt aac ttt ggt tgt gct tat
ttt gtg ttg gag aat 336Ile Lys Glu Tyr Cys Asn Phe Gly Cys Ala Tyr
Phe Val Leu Glu Asn 100 105
110cca aat cct aga gat att aaa ttt gat gat gaa aga cct cat aag cgt
384Pro Asn Pro Arg Asp Ile Lys Phe Asp Asp Glu Arg Pro His Lys Arg
115 120 125cgt aag tca aga tcc aaa tca
caa tca tca aag tca caa act aga aat 432Arg Lys Ser Arg Ser Lys Ser
Gln Ser Ser Lys Ser Gln Thr Arg Asn 130 135
140aat cgt tcc cag tca aat gcc aat gct cat ttt aca agt aaa aag cgt
480Asn Arg Ser Gln Ser Asn Ala Asn Ala His Phe Thr Ser Lys Lys Arg145
150 155 160aaa gac aca aaa
cgc cgt caa gaa cgt cat att aaa gaa gag caa gat 528Lys Asp Thr Lys
Arg Arg Gln Glu Arg His Ile Lys Glu Glu Gln Asp 165
170 175aag gaa atg acc tct gca aag cag cat ttg
tta ttc gta aga aaa aat 576Lys Glu Met Thr Ser Ala Lys Gln His Leu
Leu Phe Val Arg Lys Asn 180 185
190taa
57923192PRTStreptococcus agalactiae 23Met Arg Lys Glu Val Thr Pro Glu Met
Leu Asn Tyr Asn Lys Tyr Pro1 5 10
15Gly Pro Gln Phe Ile His Phe Glu Asn Ile Val Lys Ser Asp Asp
Ile 20 25 30Glu Phe Gln Leu
Val Ile Asn Glu Lys Ser Ala Phe Asp Val Thr Val 35
40 45Phe Gly Gln Arg Phe Ser Glu Ile Leu Leu Lys Tyr
Asp Phe Ile Val 50 55 60Gly Asp Trp
Gly Asn Glu Gln Leu Arg Leu Arg Gly Phe Tyr Lys Asp65 70
75 80Ala Ser Thr Ile Arg Lys Asn Ser
Arg Ile Ser Arg Leu Glu Asp Tyr 85 90
95Ile Lys Glu Tyr Cys Asn Phe Gly Cys Ala Tyr Phe Val Leu
Glu Asn 100 105 110Pro Asn Pro
Arg Asp Ile Lys Phe Asp Asp Glu Arg Pro His Lys Arg 115
120 125Arg Lys Ser Arg Ser Lys Ser Gln Ser Ser Lys
Ser Gln Thr Arg Asn 130 135 140Asn Arg
Ser Gln Ser Asn Ala Asn Ala His Phe Thr Ser Lys Lys Arg145
150 155 160Lys Asp Thr Lys Arg Arg Gln
Glu Arg His Ile Lys Glu Glu Gln Asp 165
170 175Lys Glu Met Thr Ser Ala Lys Gln His Leu Leu Phe
Val Arg Lys Asn 180 185
19024609DNAStreptococcus agalactiaeCDS(1)..(609) 24atg aca ata aaa aaa
gtg tta agt gta aca gga att att tta gtg aca 48Met Thr Ile Lys Lys
Val Leu Ser Val Thr Gly Ile Ile Leu Val Thr1 5
10 15gta gcg tct cta gct gct tgt agc tca aaa tct
cat act act aag acg 96Val Ala Ser Leu Ala Ala Cys Ser Ser Lys Ser
His Thr Thr Lys Thr 20 25
30ggc aaa aaa gaa gtt aat ttt gca act gtt gga aca acg gca cct ttt
144Gly Lys Lys Glu Val Asn Phe Ala Thr Val Gly Thr Thr Ala Pro Phe
35 40 45tct tat gtg aag gat ggg aaa ctg
act ggc ttt gat att gaa gta gcc 192Ser Tyr Val Lys Asp Gly Lys Leu
Thr Gly Phe Asp Ile Glu Val Ala 50 55
60aaa gct gtt ttt aaa ggt tca gat aac tat aaa gtc act ttt aaa aaa
240Lys Ala Val Phe Lys Gly Ser Asp Asn Tyr Lys Val Thr Phe Lys Lys65
70 75 80aca gaa tgg tca tcg
gta ttt acc ggc att gat tca gga aag ttt caa 288Thr Glu Trp Ser Ser
Val Phe Thr Gly Ile Asp Ser Gly Lys Phe Gln 85
90 95atg ggt gga aat aat att tct tat tca tca gag
aga tct caa aaa tay 336Met Gly Gly Asn Asn Ile Ser Tyr Ser Ser Glu
Arg Ser Gln Lys Tyr 100 105
110tta ttt tca tac cca ata ggc tct act cct tca gtt tta gca gtt cct
384Leu Phe Ser Tyr Pro Ile Gly Ser Thr Pro Ser Val Leu Ala Val Pro
115 120 125aag aat agt aat atc aaa gct
tat aat gat att agt ggt cat aaa aca 432Lys Asn Ser Asn Ile Lys Ala
Tyr Asn Asp Ile Ser Gly His Lys Thr 130 135
140cag gtt gtc caa gga acg aca act gcc aag caa tta gaa aat ttc aat
480Gln Val Val Gln Gly Thr Thr Thr Ala Lys Gln Leu Glu Asn Phe Asn145
150 155 160aaa gag cat cag
aaa aat cct gtt act cta aaa tat act aat gaa aat 528Lys Glu His Gln
Lys Asn Pro Val Thr Leu Lys Tyr Thr Asn Glu Asn 165
170 175att aca cag att cta acg aat ttg agt gat
gga aaa gct gat ttt aaa 576Ile Thr Gln Ile Leu Thr Asn Leu Ser Asp
Gly Lys Ala Asp Phe Lys 180 185
190ctt ttg acg gac caa ctg tta acg cta tta taa
609Leu Leu Thr Asp Gln Leu Leu Thr Leu Leu 195
20025202PRTStreptococcus agalactiae 25Met Thr Ile Lys Lys Val Leu Ser Val
Thr Gly Ile Ile Leu Val Thr1 5 10
15Val Ala Ser Leu Ala Ala Cys Ser Ser Lys Ser His Thr Thr Lys
Thr 20 25 30Gly Lys Lys Glu
Val Asn Phe Ala Thr Val Gly Thr Thr Ala Pro Phe 35
40 45Ser Tyr Val Lys Asp Gly Lys Leu Thr Gly Phe Asp
Ile Glu Val Ala 50 55 60Lys Ala Val
Phe Lys Gly Ser Asp Asn Tyr Lys Val Thr Phe Lys Lys65 70
75 80Thr Glu Trp Ser Ser Val Phe Thr
Gly Ile Asp Ser Gly Lys Phe Gln 85 90
95Met Gly Gly Asn Asn Ile Ser Tyr Ser Ser Glu Arg Ser Gln
Lys Tyr 100 105 110Leu Phe Ser
Tyr Pro Ile Gly Ser Thr Pro Ser Val Leu Ala Val Pro 115
120 125Lys Asn Ser Asn Ile Lys Ala Tyr Asn Asp Ile
Ser Gly His Lys Thr 130 135 140Gln Val
Val Gln Gly Thr Thr Thr Ala Lys Gln Leu Glu Asn Phe Asn145
150 155 160Lys Glu His Gln Lys Asn Pro
Val Thr Leu Lys Tyr Thr Asn Glu Asn 165
170 175Ile Thr Gln Ile Leu Thr Asn Leu Ser Asp Gly Lys
Ala Asp Phe Lys 180 185 190Leu
Leu Thr Asp Gln Leu Leu Thr Leu Leu 195
20026357DNAStreptococcus agalactiaeCDS(1)..(357) 26atg aag aat ata aca
aag cta tca act gtt gct tta agc cta cta ctt 48Met Lys Asn Ile Thr
Lys Leu Ser Thr Val Ala Leu Ser Leu Leu Leu1 5
10 15tgt acg gcg tgt gct gca tca aac acg tct aca
tct aaa aca cag tct 96Cys Thr Ala Cys Ala Ala Ser Asn Thr Ser Thr
Ser Lys Thr Gln Ser 20 25
30cat cat cct aaa caa act aaa ctc aca gat aag caa aaa gaa gaa ccc
144His His Pro Lys Gln Thr Lys Leu Thr Asp Lys Gln Lys Glu Glu Pro
35 40 45aaa aac aaa gaa gct gct gat caa
gag atg cat ccc caa ggc gct gtt 192Lys Asn Lys Glu Ala Ala Asp Gln
Glu Met His Pro Gln Gly Ala Val 50 55
60gat ttg aca aaa tat aag gca aaa ccg gtc aaa gat tat gga aaa aaa
240Asp Leu Thr Lys Tyr Lys Ala Lys Pro Val Lys Asp Tyr Gly Lys Lys65
70 75 80atc gat gtt ggt gat
ggc aag aaa atg aac att tat gaa act ggt cag 288Ile Asp Val Gly Asp
Gly Lys Lys Met Asn Ile Tyr Glu Thr Gly Gln 85
90 95gga aaa att cca att gtt ttt att cct ggt caa
gct gag att cgc cac 336Gly Lys Ile Pro Ile Val Phe Ile Pro Gly Gln
Ala Glu Ile Arg His 100 105
110gct atg ctt ata aga att taa
357Ala Met Leu Ile Arg Ile 11527118PRTStreptococcus agalactiae
27Met Lys Asn Ile Thr Lys Leu Ser Thr Val Ala Leu Ser Leu Leu Leu1
5 10 15Cys Thr Ala Cys Ala Ala
Ser Asn Thr Ser Thr Ser Lys Thr Gln Ser 20 25
30His His Pro Lys Gln Thr Lys Leu Thr Asp Lys Gln Lys
Glu Glu Pro 35 40 45Lys Asn Lys
Glu Ala Ala Asp Gln Glu Met His Pro Gln Gly Ala Val 50
55 60Asp Leu Thr Lys Tyr Lys Ala Lys Pro Val Lys Asp
Tyr Gly Lys Lys65 70 75
80Ile Asp Val Gly Asp Gly Lys Lys Met Asn Ile Tyr Glu Thr Gly Gln
85 90 95Gly Lys Ile Pro Ile Val
Phe Ile Pro Gly Gln Ala Glu Ile Arg His 100
105 110Ala Met Leu Ile Arg Ile
115281191DNAStreptococcus agalactiaeCDS(1)..(1191) 28gtg aat gaa tcg acc
atc aga aaa gaa ttt aaa ata gtt gtt ttt aaa 48Val Asn Glu Ser Thr
Ile Arg Lys Glu Phe Lys Ile Val Val Phe Lys1 5
10 15tgg atc tta aat aat caa gca gtt att gct ctc
atg att acc ttt ttg 96Trp Ile Leu Asn Asn Gln Ala Val Ile Ala Leu
Met Ile Thr Phe Leu 20 25
30gta ttt tta acg att ttt att ttt acc aaa atc tct ttt atg ttt aaa
144Val Phe Leu Thr Ile Phe Ile Phe Thr Lys Ile Ser Phe Met Phe Lys
35 40 45cct gtg ttt gat ttt ctt gct gtg
ctg ata ttg ccg ctt gta att tct 192Pro Val Phe Asp Phe Leu Ala Val
Leu Ile Leu Pro Leu Val Ile Ser 50 55
60ggc ttg ctt tat tac cta tta aaa cct atg gtt aca ttt tta gag aag
240Gly Leu Leu Tyr Tyr Leu Leu Lys Pro Met Val Thr Phe Leu Glu Lys65
70 75 80cgg gga att aag cgt
gta aca gcg ata tta tca gtt ttt act att ata 288Arg Gly Ile Lys Arg
Val Thr Ala Ile Leu Ser Val Phe Thr Ile Ile 85
90 95atc ctt ctg tta att tgg gca atg tct agt ttt
att ccc atg atg agt 336Ile Leu Leu Leu Ile Trp Ala Met Ser Ser Phe
Ile Pro Met Met Ser 100 105
110aat caa tta cgc cat ttt atg gaa gat ctc cct tca tat gtg aat aaa
384Asn Gln Leu Arg His Phe Met Glu Asp Leu Pro Ser Tyr Val Asn Lys
115 120 125gtg caa atg gaa aca agt tcg
ttt ata gat cac aac cct tgg tta aaa 432Val Gln Met Glu Thr Ser Ser
Phe Ile Asp His Asn Pro Trp Leu Lys 130 135
140tct tat aaa ggg gaa ata tcg agc atg tta tct aat atc agt agc caa
480Ser Tyr Lys Gly Glu Ile Ser Ser Met Leu Ser Asn Ile Ser Ser Gln145
150 155 160gcg gtc tct tat
gct gaa aaa ttt tca aag aat gtt tta gat tgg gca 528Ala Val Ser Tyr
Ala Glu Lys Phe Ser Lys Asn Val Leu Asp Trp Ala 165
170 175gga aat tta gct agt aca gtt gca cgt gtg
aca gta gca aca atc atg 576Gly Asn Leu Ala Ser Thr Val Ala Arg Val
Thr Val Ala Thr Ile Met 180 185
190gct ccc ttt att ttg ttt tat ctt tta aga gat agt cgc aac atg aag
624Ala Pro Phe Ile Leu Phe Tyr Leu Leu Arg Asp Ser Arg Asn Met Lys
195 200 205aat ggt ttc tta atg gtt tta
cca acc aaa cta cgc caa cca gct gat 672Asn Gly Phe Leu Met Val Leu
Pro Thr Lys Leu Arg Gln Pro Ala Asp 210 215
220cgt att ttg cga gaa atg aat agt caa atg tca gga tat gtg caa gga
720Arg Ile Leu Arg Glu Met Asn Ser Gln Met Ser Gly Tyr Val Gln Gly225
230 235 240caa atc att gtt
gct att act gtt ggt gtt att ttt tca ata atg tat 768Gln Ile Ile Val
Ala Ile Thr Val Gly Val Ile Phe Ser Ile Met Tyr 245
250 255agt att ata ggc ctt aga tat ggc gtg aca
tta ggg att att gcc ggt 816Ser Ile Ile Gly Leu Arg Tyr Gly Val Thr
Leu Gly Ile Ile Ala Gly 260 265
270gtg tta aat atg gtt ccc tat ttg gga agt ttt gtc gcc caa att cca
864Val Leu Asn Met Val Pro Tyr Leu Gly Ser Phe Val Ala Gln Ile Pro
275 280 285gtg ttt atc tta gcg ctt gtc
gca gga cct gtt atg gtt gtt aaa gtt 912Val Phe Ile Leu Ala Leu Val
Ala Gly Pro Val Met Val Val Lys Val 290 295
300gcg att gtt ttt gtt att gag caa act cta gag gga cgc ttt gtc tca
960Ala Ile Val Phe Val Ile Glu Gln Thr Leu Glu Gly Arg Phe Val Ser305
310 315 320cct ttg gtt tta
ggt aat aaa ctt agc att cat cca att aca att atg 1008Pro Leu Val Leu
Gly Asn Lys Leu Ser Ile His Pro Ile Thr Ile Met 325
330 335ttt att tta tta acc tct gga gcg atg ttt
ggt gtt tgg gga gta ttc 1056Phe Ile Leu Leu Thr Ser Gly Ala Met Phe
Gly Val Trp Gly Val Phe 340 345
350ctc agt att ccg att tat gca tct atc aaa gtt gtt gtt aaa gaa ttg
1104Leu Ser Ile Pro Ile Tyr Ala Ser Ile Lys Val Val Val Lys Glu Leu
355 360 365ttt gat tgg tac aaa gct gtc
agt ggg cta tat aca ata gat gtt gtt 1152Phe Asp Trp Tyr Lys Ala Val
Ser Gly Leu Tyr Thr Ile Asp Val Val 370 375
380act gaa gaa aga agt gaa gaa gtt aaa aat gtt gaa tag
1191Thr Glu Glu Arg Ser Glu Glu Val Lys Asn Val Glu385
390 39529396PRTStreptococcus agalactiae 29Val Asn Glu Ser
Thr Ile Arg Lys Glu Phe Lys Ile Val Val Phe Lys1 5
10 15Trp Ile Leu Asn Asn Gln Ala Val Ile Ala
Leu Met Ile Thr Phe Leu 20 25
30Val Phe Leu Thr Ile Phe Ile Phe Thr Lys Ile Ser Phe Met Phe Lys
35 40 45Pro Val Phe Asp Phe Leu Ala Val
Leu Ile Leu Pro Leu Val Ile Ser 50 55
60Gly Leu Leu Tyr Tyr Leu Leu Lys Pro Met Val Thr Phe Leu Glu Lys65
70 75 80Arg Gly Ile Lys Arg
Val Thr Ala Ile Leu Ser Val Phe Thr Ile Ile 85
90 95Ile Leu Leu Leu Ile Trp Ala Met Ser Ser Phe
Ile Pro Met Met Ser 100 105
110Asn Gln Leu Arg His Phe Met Glu Asp Leu Pro Ser Tyr Val Asn Lys
115 120 125Val Gln Met Glu Thr Ser Ser
Phe Ile Asp His Asn Pro Trp Leu Lys 130 135
140Ser Tyr Lys Gly Glu Ile Ser Ser Met Leu Ser Asn Ile Ser Ser
Gln145 150 155 160Ala Val
Ser Tyr Ala Glu Lys Phe Ser Lys Asn Val Leu Asp Trp Ala
165 170 175Gly Asn Leu Ala Ser Thr Val
Ala Arg Val Thr Val Ala Thr Ile Met 180 185
190Ala Pro Phe Ile Leu Phe Tyr Leu Leu Arg Asp Ser Arg Asn
Met Lys 195 200 205Asn Gly Phe Leu
Met Val Leu Pro Thr Lys Leu Arg Gln Pro Ala Asp 210
215 220Arg Ile Leu Arg Glu Met Asn Ser Gln Met Ser Gly
Tyr Val Gln Gly225 230 235
240Gln Ile Ile Val Ala Ile Thr Val Gly Val Ile Phe Ser Ile Met Tyr
245 250 255Ser Ile Ile Gly Leu
Arg Tyr Gly Val Thr Leu Gly Ile Ile Ala Gly 260
265 270Val Leu Asn Met Val Pro Tyr Leu Gly Ser Phe Val
Ala Gln Ile Pro 275 280 285Val Phe
Ile Leu Ala Leu Val Ala Gly Pro Val Met Val Val Lys Val 290
295 300Ala Ile Val Phe Val Ile Glu Gln Thr Leu Glu
Gly Arg Phe Val Ser305 310 315
320Pro Leu Val Leu Gly Asn Lys Leu Ser Ile His Pro Ile Thr Ile Met
325 330 335Phe Ile Leu Leu
Thr Ser Gly Ala Met Phe Gly Val Trp Gly Val Phe 340
345 350Leu Ser Ile Pro Ile Tyr Ala Ser Ile Lys Val
Val Val Lys Glu Leu 355 360 365Phe
Asp Trp Tyr Lys Ala Val Ser Gly Leu Tyr Thr Ile Asp Val Val 370
375 380Thr Glu Glu Arg Ser Glu Glu Val Lys Asn
Val Glu385 390 395301230DNAStreptococcus
agalactiaeCDS(1)..(1230) 30atg ttt atg gga atc cca caa tat ttc ttc tac
ctt atc tta gct gtc 48Met Phe Met Gly Ile Pro Gln Tyr Phe Phe Tyr
Leu Ile Leu Ala Val1 5 10
15cta cca att tac atc ggc tta ttc ttt aag aag cgt ttt gcc tta tat
96Leu Pro Ile Tyr Ile Gly Leu Phe Phe Lys Lys Arg Phe Ala Leu Tyr
20 25 30gag att att ttt agt cta agt
ttt att gta atg atg ttg act ggt agt 144Glu Ile Ile Phe Ser Leu Ser
Phe Ile Val Met Met Leu Thr Gly Ser 35 40
45act ttt aat caa ttg aag tca cta ttg gca tac gtt gtc gga cag
tct 192Thr Phe Asn Gln Leu Lys Ser Leu Leu Ala Tyr Val Val Gly Gln
Ser 50 55 60ctg cta gtt ttt atc tat
aaa gct tac cgg aaa cga ttt aat cat act 240Leu Leu Val Phe Ile Tyr
Lys Ala Tyr Arg Lys Arg Phe Asn His Thr65 70
75 80ttg gtc ttt tat gta acg gtt tgt tta tct att
ttt ccg cta ttt ttg 288Leu Val Phe Tyr Val Thr Val Cys Leu Ser Ile
Phe Pro Leu Phe Leu 85 90
95gta aaa tta att cca gct ata tct gag gat ggg cat cag tca ctt ttt
336Val Lys Leu Ile Pro Ala Ile Ser Glu Asp Gly His Gln Ser Leu Phe
100 105 110ggg ttt cta gga att tct
tac ctt act ttt aga gct gtt gct atg att 384Gly Phe Leu Gly Ile Ser
Tyr Leu Thr Phe Arg Ala Val Ala Met Ile 115 120
125att gaa atg aga gac ggt gtc ttg aaa gaa ttt act tta tgg
gaa ttc 432Ile Glu Met Arg Asp Gly Val Leu Lys Glu Phe Thr Leu Trp
Glu Phe 130 135 140tta aga ttt tta ctc
ttc ttt cca act ttc tca agt gga cca att gat 480Leu Arg Phe Leu Leu
Phe Phe Pro Thr Phe Ser Ser Gly Pro Ile Asp145 150
155 160cgt ttt aaa cga ttc aat gag gat tac att
aat atc cca gat cga aac 528Arg Phe Lys Arg Phe Asn Glu Asp Tyr Ile
Asn Ile Pro Asp Arg Asn 165 170
175gaa ctc cta gat atg tta ggt caa gcg att cat tat ttg atg tta ggt
576Glu Leu Leu Asp Met Leu Gly Gln Ala Ile His Tyr Leu Met Leu Gly
180 185 190ttt ctc tat aag ttt att
tta gcc tat att ttt gga agt ctg att atg 624Phe Leu Tyr Lys Phe Ile
Leu Ala Tyr Ile Phe Gly Ser Leu Ile Met 195 200
205cct cct cta aaa gaa tta gcg cta gaa cag ggt ggt gtg ttt
aat tgg 672Pro Pro Leu Lys Glu Leu Ala Leu Glu Gln Gly Gly Val Phe
Asn Trp 210 215 220cca aca ctt ggg gtt
atg tat gcc ttt ggt ttt gat ttg ttc ttt gat 720Pro Thr Leu Gly Val
Met Tyr Ala Phe Gly Phe Asp Leu Phe Phe Asp225 230
235 240ttt gca ggt tac aca atg ttt gcg ttg gct
att tct aac cta atg ggg 768Phe Ala Gly Tyr Thr Met Phe Ala Leu Ala
Ile Ser Asn Leu Met Gly 245 250
255att aag tct ccg att aac ttt gac aaa cct ttc aaa tca cgc gac cta
816Ile Lys Ser Pro Ile Asn Phe Asp Lys Pro Phe Lys Ser Arg Asp Leu
260 265 270aaa gaa ttt tgg aat aga
tgg cat atg agc ctt tct ttc tgg ttt aga 864Lys Glu Phe Trp Asn Arg
Trp His Met Ser Leu Ser Phe Trp Phe Arg 275 280
285gac ttt gtt ttc atg agg ctt gtt aag ctt tta gtt aaa aat
aaa gtt 912Asp Phe Val Phe Met Arg Leu Val Lys Leu Leu Val Lys Asn
Lys Val 290 295 300ttt aaa aac cgt aat
gtt act tca agt gta gct tat att atc aat atg 960Phe Lys Asn Arg Asn
Val Thr Ser Ser Val Ala Tyr Ile Ile Asn Met305 310
315 320ctt ctt atg gga ttc tgg cat ggg tta act
tgg tac tat ata gcc tat 1008Leu Leu Met Gly Phe Trp His Gly Leu Thr
Trp Tyr Tyr Ile Ala Tyr 325 330
335ggt ctc ttt cat ggg att ggc cta gtt att aat gac gct tgg gta cgt
1056Gly Leu Phe His Gly Ile Gly Leu Val Ile Asn Asp Ala Trp Val Arg
340 345 350aag aag aaa aat ayt aat
aaa gaa aga aga ttg gct aaa aaa cca ctt 1104Lys Lys Lys Asn Xaa Asn
Lys Glu Arg Arg Leu Ala Lys Lys Pro Leu 355 360
365tta cca gaa aac aaa tgg act tat gct ttg ggt gtc ttc atc
acc ttt 1152Leu Pro Glu Asn Lys Trp Thr Tyr Ala Leu Gly Val Phe Ile
Thr Phe 370 375 380aat gta gtt atg ttt
tct ttc ttg att ttt tca gga ttt tta gat ctt 1200Asn Val Val Met Phe
Ser Phe Leu Ile Phe Ser Gly Phe Leu Asp Leu385 390
395 400ttg tgg ttc cca caa ccg cat aac aaa taa
1230Leu Trp Phe Pro Gln Pro His Asn Lys
40531409PRTStreptococcus agalactiaemisc_feature(357)..(357)The
'Xaa' at location 357 stands for Thr, or Ile. 31Met Phe Met Gly Ile
Pro Gln Tyr Phe Phe Tyr Leu Ile Leu Ala Val1 5
10 15Leu Pro Ile Tyr Ile Gly Leu Phe Phe Lys Lys
Arg Phe Ala Leu Tyr 20 25
30Glu Ile Ile Phe Ser Leu Ser Phe Ile Val Met Met Leu Thr Gly Ser
35 40 45Thr Phe Asn Gln Leu Lys Ser Leu
Leu Ala Tyr Val Val Gly Gln Ser 50 55
60Leu Leu Val Phe Ile Tyr Lys Ala Tyr Arg Lys Arg Phe Asn His Thr65
70 75 80Leu Val Phe Tyr Val
Thr Val Cys Leu Ser Ile Phe Pro Leu Phe Leu 85
90 95Val Lys Leu Ile Pro Ala Ile Ser Glu Asp Gly
His Gln Ser Leu Phe 100 105
110Gly Phe Leu Gly Ile Ser Tyr Leu Thr Phe Arg Ala Val Ala Met Ile
115 120 125Ile Glu Met Arg Asp Gly Val
Leu Lys Glu Phe Thr Leu Trp Glu Phe 130 135
140Leu Arg Phe Leu Leu Phe Phe Pro Thr Phe Ser Ser Gly Pro Ile
Asp145 150 155 160Arg Phe
Lys Arg Phe Asn Glu Asp Tyr Ile Asn Ile Pro Asp Arg Asn
165 170 175Glu Leu Leu Asp Met Leu Gly
Gln Ala Ile His Tyr Leu Met Leu Gly 180 185
190Phe Leu Tyr Lys Phe Ile Leu Ala Tyr Ile Phe Gly Ser Leu
Ile Met 195 200 205Pro Pro Leu Lys
Glu Leu Ala Leu Glu Gln Gly Gly Val Phe Asn Trp 210
215 220Pro Thr Leu Gly Val Met Tyr Ala Phe Gly Phe Asp
Leu Phe Phe Asp225 230 235
240Phe Ala Gly Tyr Thr Met Phe Ala Leu Ala Ile Ser Asn Leu Met Gly
245 250 255Ile Lys Ser Pro Ile
Asn Phe Asp Lys Pro Phe Lys Ser Arg Asp Leu 260
265 270Lys Glu Phe Trp Asn Arg Trp His Met Ser Leu Ser
Phe Trp Phe Arg 275 280 285Asp Phe
Val Phe Met Arg Leu Val Lys Leu Leu Val Lys Asn Lys Val 290
295 300Phe Lys Asn Arg Asn Val Thr Ser Ser Val Ala
Tyr Ile Ile Asn Met305 310 315
320Leu Leu Met Gly Phe Trp His Gly Leu Thr Trp Tyr Tyr Ile Ala Tyr
325 330 335Gly Leu Phe His
Gly Ile Gly Leu Val Ile Asn Asp Ala Trp Val Arg 340
345 350Lys Lys Lys Asn Xaa Asn Lys Glu Arg Arg Leu
Ala Lys Lys Pro Leu 355 360 365Leu
Pro Glu Asn Lys Trp Thr Tyr Ala Leu Gly Val Phe Ile Thr Phe 370
375 380Asn Val Val Met Phe Ser Phe Leu Ile Phe
Ser Gly Phe Leu Asp Leu385 390 395
400Leu Trp Phe Pro Gln Pro His Asn Lys
40532100DNAStreptococcus agalactiaeCDS(1)..(99) 32atg aat aaa ata acg aca
tta tca acc atc gcc ctg act tta atg ctt 48Met Asn Lys Ile Thr Thr
Leu Ser Thr Ile Ala Leu Thr Leu Met Leu1 5
10 15tgc gtt gga tgt tct gcc aat aaa gat aat caa aaa
act aaa act gag 96Cys Val Gly Cys Ser Ala Asn Lys Asp Asn Gln Lys
Thr Lys Thr Glu 20 25 30gat
c
100Asp3333PRTStreptococcus agalactiae 33Met Asn Lys Ile Thr Thr Leu Ser
Thr Ile Ala Leu Thr Leu Met Leu1 5 10
15Cys Val Gly Cys Ser Ala Asn Lys Asp Asn Gln Lys Thr Lys
Thr Glu 20 25
30Asp34654DNAStreptococcus agalactiaeCDS(1)..(654) 34gat cga ggc tat caa
gaa gca atg gct aaa cta agg aaa act tac ggc 48Asp Arg Gly Tyr Gln
Glu Ala Met Ala Lys Leu Arg Lys Thr Tyr Gly1 5
10 15gaa tat ggt tta ggg gtt tct aca gga tta gat
tta cct gaa tca gaa 96Glu Tyr Gly Leu Gly Val Ser Thr Gly Leu Asp
Leu Pro Glu Ser Glu 20 25
30ggt tat gta cct gga aaa tac agc tta gga aca act cta atg gaa tcg
144Gly Tyr Val Pro Gly Lys Tyr Ser Leu Gly Thr Thr Leu Met Glu Ser
35 40 45ttc ggt cag tat gat gcc tat aca
cca atg caa ctt ggt cag tat atc 192Phe Gly Gln Tyr Asp Ala Tyr Thr
Pro Met Gln Leu Gly Gln Tyr Ile 50 55
60tca act att gcg aat aat ggg aat cgt tta gca cct cac gtg gtt tca
240Ser Thr Ile Ala Asn Asn Gly Asn Arg Leu Ala Pro His Val Val Ser65
70 75 80gat atc tat gaa ggg
aat gat tct aat aag ttc gct caa ttg gtt cgt 288Asp Ile Tyr Glu Gly
Asn Asp Ser Asn Lys Phe Ala Gln Leu Val Arg 85
90 95tca atc act cct aaa aca cta aat aag ata gct
atc tca gat caa gag 336Ser Ile Thr Pro Lys Thr Leu Asn Lys Ile Ala
Ile Ser Asp Gln Glu 100 105
110tta gcc att att caa gaa ggt ttt tat aac gtt gtc aat agt gga agt
384Leu Ala Ile Ile Gln Glu Gly Phe Tyr Asn Val Val Asn Ser Gly Ser
115 120 125ggc tat gca act ggt acg tca
atg agg ggg aat gtg aca acc att agy 432Gly Tyr Ala Thr Gly Thr Ser
Met Arg Gly Asn Val Thr Thr Ile Ser 130 135
140ggt aaa act ggt acc gct gaa aca ttt gct aaa aat ata aat gga caa
480Gly Lys Thr Gly Thr Ala Glu Thr Phe Ala Lys Asn Ile Asn Gly Gln145
150 155 160aca gtt tct acc
tac aac tta aac gct att gcc tac gat act aat cgt 528Thr Val Ser Thr
Tyr Asn Leu Asn Ala Ile Ala Tyr Asp Thr Asn Arg 165
170 175aaa ata gca gta gcg gta atg tat ccg cat
gtt aca act gat aca aca 576Lys Ile Ala Val Ala Val Met Tyr Pro His
Val Thr Thr Asp Thr Thr 180 185
190aaa tcc cat caa tta gtt gca cga gat atg att gat caa tat att tca
624Lys Ser His Gln Leu Val Ala Arg Asp Met Ile Asp Gln Tyr Ile Ser
195 200 205cag tca cag gac aat aag aga
gga cat tga 654Gln Ser Gln Asp Asn Lys Arg
Gly His 210 21535217PRTStreptococcus agalactiae 35Asp
Arg Gly Tyr Gln Glu Ala Met Ala Lys Leu Arg Lys Thr Tyr Gly1
5 10 15Glu Tyr Gly Leu Gly Val Ser
Thr Gly Leu Asp Leu Pro Glu Ser Glu 20 25
30Gly Tyr Val Pro Gly Lys Tyr Ser Leu Gly Thr Thr Leu Met
Glu Ser 35 40 45Phe Gly Gln Tyr
Asp Ala Tyr Thr Pro Met Gln Leu Gly Gln Tyr Ile 50 55
60Ser Thr Ile Ala Asn Asn Gly Asn Arg Leu Ala Pro His
Val Val Ser65 70 75
80Asp Ile Tyr Glu Gly Asn Asp Ser Asn Lys Phe Ala Gln Leu Val Arg
85 90 95Ser Ile Thr Pro Lys Thr
Leu Asn Lys Ile Ala Ile Ser Asp Gln Glu 100
105 110Leu Ala Ile Ile Gln Glu Gly Phe Tyr Asn Val Val
Asn Ser Gly Ser 115 120 125Gly Tyr
Ala Thr Gly Thr Ser Met Arg Gly Asn Val Thr Thr Ile Ser 130
135 140Gly Lys Thr Gly Thr Ala Glu Thr Phe Ala Lys
Asn Ile Asn Gly Gln145 150 155
160Thr Val Ser Thr Tyr Asn Leu Asn Ala Ile Ala Tyr Asp Thr Asn Arg
165 170 175Lys Ile Ala Val
Ala Val Met Tyr Pro His Val Thr Thr Asp Thr Thr 180
185 190Lys Ser His Gln Leu Val Ala Arg Asp Met Ile
Asp Gln Tyr Ile Ser 195 200 205Gln
Ser Gln Asp Asn Lys Arg Gly His 210 215
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