Abstract:
The invention relates to  Streptococcus suis  infection in pigs, vaccines directed against those infections and tests for diagnosing  Streptococcus suis  infections. The invention provides an isolated or recombinant nucleic acid encoding a capsular gene cluster of  Streptococcus suis  or a gene or gene fragment derivated thereof. The invention further provides a nucleic acid probe or primer allowing species or serotype specific detection of  Streptococcus suis.  The invention also provides a  Streptococcus suis  antigen and vaccine derived thereof.

Description:
CROSS-REFERENCE TO RELATED APPLICATIONS  
       [0001]    This application claims priority to, and is a continuation of, International Application No. PCT/NL99/00460, filed on Jul. 19, 1999, designating the United States of America, the contents of which are incorporated herein by this reference, the PCT International Patent Application itself claiming priority from European Patent Office Application Ser. No. 98202465.5 filed Jul. 22, 1998 and European Patent Office Application Ser. No. 98202467.1 filed Jul. 22, 1998. 
     
    
     
       TECHNICAL FIELD  
         [0002]    The invention relates to  Streptococcus  infections in pigs, vaccines directed against those infections, tests for diagnosing  Streptococcus  infections and bacterial vaccines. More particularly, the invention relates to vaccines directed against  Streptococcus  infections.  
         BACKGROUND OF THE INVENTION  
         [0003]    [0003] Streptococcus species,  of which a large variety cause infections in domestic animals and man, are often grouped according to Lancefield&#39;s groups. Typing according to Lancefield occurs on the basis of serological determinants or antigens that are, among others, present in the capsule of the bacterium, and allows for only an approximate determination. Often, bacteria from different groups show cross-reactivity with each other, while other Streptococci cannot be assigned a group-determinant at all. Within groups, further differentiation is often possible on the basis of serotyping. These serotypes further contribute to the large antigenic variability of Streptococci, a fact that creates an array of difficulties within diagnosis of and vaccination against Streptococcal infections.  
           [0004]    Lancefield group A  Streptococcus species  (Group A streptococci “GAS”,  Streptococcus pyogenes ) are common in children, causing nasopharyngeal infections and complications thereof. Among animals, cattle are especially susceptible to GAS, and the resulting mastitis.  
           [0005]    Group A streptococci are the etiologic agents of streptococcal pharyngitis and impetigo, two of the most common bacterial infections in children, as well as a variety of less common, but potentially life-threatening, infections including soft tissue infections, bacteremia, and pneumonia. In addition, GAS are uniquely associated with the post-infectious autoimmune syndromes of acute rheumatic fever and post streptococcal glomerulonephritis.  
           [0006]    Several recent reports suggest that the incidence of both serious infections due to GAS and acute rheumatic fever has increased during the past decade, focusing renewed interest on defining the attributes or virulence factors of the organism that may play a role in the pathogenesis of these diseases.  
           [0007]    GAS produce several surface components and extracellular products that may be important in virulence. The major surface protein, M protein, has been studied in the most detail and has been convincingly shown to play a role in both virulence and immunity. Isolates rich in M protein are able to grow in human blood, a property thought to reflect the capacity of M protein to interfere with phagocytosis, and these isolates tend to be virulent in experimental animals.  
           [0008]    Lancefield group B  Streptococcus  (“GBS”) are most often seen in cattle, causing mastitis; however, human infants are susceptible as well, often with fatal consequences. Group B streptococci (GBS) constitute a major cause of bacterial sepsis and meningitis among human neonates born in the United States and Western Europe and are emerging as significant neonatal pathogens in developing countries as well.  
           [0009]    It is estimated that GBS strains are responsible for 10,000 to 15,000 cases of invasive infection in neonates in the United States alone. Despite advances in early diagnosis and treatment, neonatal sepsis due to GBS continues to carry a mortality rate of 15 to 20%. In addition, survivors of GBS meningitis have 30 to 50% incidence of long-term neurologic sequelae. Over the past two decades, increasing recognition of GBS as an important pathogen for human infants has generated renewed interest in defining the bacterial and host factors important in virulence of GBS and in the immune response to GBS infection.  
           [0010]    Particular attention has focused on the capsular polysaccharide as the predominant surface antigen of the organisms. In a modification of the system originally developed by Rebecca Lancefield, GBS strains are serotyped on the basis of antigenic differences in their capsular polysaccharides and the presence or absence of serologically defined C proteins. While GBS isolated from nonhuman sources often lack a serologically detectable capsule, a large majority of strains associated with neonatal infection belong to one of four major capsular serotypes, 1 a , 1 b , II or III. The capsular polysaccharide forms the outermost layer around the exterior of the bacterial cell, superficial to the cell wall. The capsule is distinct from the cell wall-associated group B carbohydrate. It has been suggested that the presence of sialic acid, in the capsule of bacteria that causes meningitis, is important for allowing these bacteria to breach the blood-brain barrier. Indeed, in  S. agalactiae,  sialic acid has been shown to be critical for the virulence function of the type III capsule. The capsule of  S. suis  serotype is composed of glucose, galactose, N-acetylglucosamine, rhamnose and sialic acid.  
           [0011]    The group B polysaccharide, in contrast to the type-specific capsule, is present on all GBS strains and is the basis for serogrouping the organisms into Lancefield&#39;s group B. Early studies by Lancefield and co-workers showed that antibodies raised in rabbits against whole GBS organisms protected mice against challenge with strains of homologous capsular type, demonstrating the central role of the capsular polysaccharide as a protective antigen. Studies in the 1970s by Baker and Kasper demonstrated that cord blood of human infants with type III GBS sepsis uniformly had low or undetectable levels of antibodies directed against the type III capsule, suggesting that a deficiency of anticapsular antibody was a key factor in susceptibility of human neonates to GBS disease.  
           [0012]    Lancefield group C infections, such as those with  S. equi, S. zooepidemicus, S. dysgalactiae,  and others, are mainly seen in horses, cattle and pigs, but can also cross the species barrier to humans. Lancefield group D ( S. bovis ) infections are found in all mammals and some birds, sometimes resulting in endocarditis or septicemia.  
           [0013]    Lancefield groups E, G, L, P, U and V ( S. porcinus, S. canis, S. dysgalactiae ) are found in various hosts, causing neonatal infections, nasopharyngeal infections or mastitis.  
           [0014]    Within Lancefield groups R, S, and T (and with ungrouped types),  Streptococcus suis  is an important cause of meningitis, septicemia, arthritis and sudden death in young pigs (4, 46). Incidentally, it can also cause meningitis in man (1).  S. suis  strains are usually identified and classified by their morphological, biochemical and serological characteristics (58, 59, 46). Serological classification is based on the presence of specific antigenic polysaccharides. So far, 35 different serotypes have been described (9, 56, 14). In several European countries,  S. suis  serotype 2 is the most prevalent type isolated from diseased pigs, followed by serotypes 9 and 1. Serological typing of  S. suis  is performed using different types of agglutination tests. In these tests, isolated and biochemically characterized  S. suis  cells are agglutinated with a panel of 35 specific sera. These methods are very laborious and time-consuming.  
           [0015]    Little is known about the pathogenesis of the disease caused by  S. suis,  let alone about its various serotypes such as type 2. Various bacterial components, such as extracellular and cell-membrane associated proteins, fimbriae, hemagglutinins, and hemolysin have been suggested as virulence factors (9, 10, 11, 15, 16, 47, 49). However, the precise role of these protein components in the pathogenesis of the disease remains unclear (37). It is well known that the polysaccharide capsule of various Streptococci and other Gram-positive bacteria plays an important role in pathogenesis (3, 6, 35, 51, 52). The capsule enables these microorganisms to resist phagocytosis and is therefore regarded as an important virulence factor. Recently, a role of the capsule of  S. suis  in the pathogenesis was suggested as well (5). However, the structure, organization and function of the genes responsible for capsule polysaccharide synthesis (“ cps ”) in  S. suis  is unknown. Within S. suis, serotype 1 and 2, strains can differ in virulence for pigs (41, 45, 49). Some type 1 and 2 strains are virulent, other strains are not. Because both virulent and nonvirulent strains of serotype 1 and 2 strains are fully encapsulated, it may even be that the capsule is not a relevant factor required for virulence.  
           [0016]    Attempts to control  S. suis  infections or disease are still hampered by the lack of knowledge about the epidemiology of the disease and the lack of effective vaccines and sensitive diagnostics. It is well known and generally accepted that the polysaccharide capsule of various Streptococci and other gram-positive bacteria plays an important role in pathogenesis. The capsule enables these microorganisms to resist phagocytosis and is therefore regarded as an important virulence factor.  
           [0017]    Compared to encapsulated  S. suis  strains, non-encapsulated  S. suis  strains are phagocytosed by murine polymorphonuclear leucocytes to a greater degree. Moreover, an increase in thickness of capsule was noted for in vivo grown virulent strains while no increase was observed for avirulent strains. Therefore, these data again demonstrate the role of the capsule in the pathogenesis for  S. suis  as well. Ungrouped  Streptoccus species,  such as  S. mutans,  causing caries in humans,  S. uberis.  causing mastitis in cattle, and  S. pneumonia,  causing major infections in humans, and  Enterococcusfaecilalis  and  E. faecium,  further contribute to the large group of Streptococci.  
           [0018]    [0018] Streptococcus pneumoniae  (the pneumococcus) is a human pathogen causing invasive diseases, such as pneumonia, bacteremia, and meningitis. Despite the availability of antibiotics, pneumococcal infections remain common and can still be fatal, especially in high-risk groups, such as young children and elderly people. Particularly in developing countries, many children under the age of five years die each year from pneumococcal pneumonia.  S. pneumoniae  is also the leading cause of otitis media and sinusitis. These infections are less serious, but nevertheless incur substantial medical costs, especially when leading to complications, such as permanent deafness. The normal ecological niche of the pneumococcus is the nasopharynx of man. The entire human population is colonized by the pneumococcus at one time or another, and at a given time, up to 60% of individuals may be carriers. Nasopharyngeal carriage of pneumococci by man is often accompanied by the development of protection against infection by the same serotype. Most infections do not occur after prolonged carriage but follow exposure to recently acquired strains. Many bacteria contain surface polysaccharides that act as a protective layer against the environment. Surface polysaccharides of pathogenic bacteria usually make the bacteria resistant to the defense mechanisms of the host, for example, the lytic action of serum or phagocytosis. In this respect, the serotype-specific capsular polysaccharide (° CP”) of  Streptococcus pneumoniae,  is an important virulence factor. Unencapsulated strains are avirulent, and antibodies directed against the CP are protective. Protection is serotype specific; each serotype has its own, specific CP structure. Ninety different capsular serotypes have been identified. Currently, CPs of 23 serotypes are included in a vaccine.  
           [0019]    Vaccines directed against  Streptococcus  infections typically aim to utilize an immune response directed against the polysaccharide capsule of the various  Streptococcus species,  especially since the capsule is considered a primary virulence factor for these bacteria. During infection, the capsule provides resistance against phagocytosis and thus protects the bacteria from the immune system of the host, and from elimination by macrophages and neutrophils.  
           [0020]    The capsule particularly confers the bacterium resistance to complement-mediated opsonophagocytosis. In addition, some bacteria express capsular polysaccharides (CPs) that mimic host molecules, thereby avoiding the immune system of the host. Also, even when the bacteria have been phagocytosed, intracellular killing is hampered by the presence of a capsule.  
           [0021]    It is generally thought that the bacterium will be recognized by the immune system through the anticapsular-antibodies or serum-factors bound to its capsule, and will, through opsonization, be phagocytosed and killed only when the host has antibodies or other serum factors directed against capsule antigens.  
           [0022]    However, these antibodies are serotype-specific, and will often only confer protection against only one of the many serotypes known within a group of  Streptococci.    
           [0023]    For example, current commercially available  S. suis  vaccines, which are generally based on whole-cell-bacterial preparations, or on capsule-enriched fractions of  S. suis,  confer only limited protection against heterologous strains. Also, the current pneumococcal vaccine, which was licensed in the United states in 1983, consists of purified CPs of 23 pneumococcal serotypes whereas at least 90 CP types exist.  
           [0024]    The composition of this pneumococcal vaccine was based on the frequency of the occurrence of disease isolates in the US and cross-reactivity between various serotypes. Although this vaccine protects healthy adults against infections caused by serotypes included in the vaccine, it fails to raise a protective immune response in infants younger than 18 months and it is less effective in elderly people. In addition, the vaccine confers only limited protection in patients with immunodeficiencies and hematology malignancies.  
           [0025]    Thus, improved vaccines are needed against  Streptococcus  infections. Much attention is directed toward producing CP vaccines by producing the relevant polysaccharides via chemical or recombinant means. However, chemical synthesis of polysaccharides is costly, and capsular polysaccharide synthesis by recombinant means necessitates knowledge about the relevant genes, which is not always available, and needs to be determined for every relevant serotype.  
         DISCLOSURE OF THE INVENTION  
         [0026]    The invention provides an isolated or recombinant nucleic acid encoding a capsular ( cps ) gene cluster of  Streptococcus suis.  Biosynthesis of capsule polysaccharides has generally been studied in a number of Gram-positive and Gram-negative bacteria (32). In Gram-negative bacteria, but also in a number of Gram-positive bacteria, genes which are involved in the biosynthesis of polysaccharides are clustered at a single locus.  
           [0027]    [0027] Streptococcus suis  capsular genes, as provided by the invention, show a common genetic organization involving three distinct regions. The central region is serotype specific and encodes enzymes responsible for the synthesis and polymerization of the polysaccharides. The central region is flanked by two regions conserved in  Streptococcus suis  which encode proteins for common functions, such as transport of the polysaccharide across the cellular membrane. However, between species, only low homologies exist, hampering easy comparison and detection of seemingly similar genes. Knowing the nucleic acid encoding the flanking regions allows type-specific determination of nucleic acid of the central region of  Streptococcus suis  serotypes, as, for example, described herein.  
           [0028]    The invention provides an isolated or recombinant nucleic acid encoding a capsular gene cluster of  Streptococcus suis  or a gene or gene fragment derived thereof. Such a nucleic acid is, for example, provided by hybridizing chromosomal DNA derived from any one of the  Streptococcus suis  serotypes to a nucleic acid encoding a gene derived from a  Streptococcus suis  serotype 1, 2 or 9 capsular gene cluster, as provided by the invention (see for example, Tables 4 and 5) and cloning of (type-specific) genes as, for example, described herein. At least 14 open reading frames are identified. Most of the genes belong to a single transcriptional unit, identifying a coordinate control of these genes. The genes and the enzymes and proteins they encode, act in concert to provide the capsule with the relevant polysaccharides.  
           [0029]    The invention provides cps genes and proteins encoded thereof involved in regulation (CpsA), chain length determination (CpsB, C), export (CpsC) and biosynthesis (CpsE, F, G, H, J, K). Although, at first glance, the overall organization seemed to be similar to that of the  cps  and  eps  gene clusters of a number of Gram-positive bacteria (19, 32, 42), overall homologies are low (see, table 3). The region involved in biosynthesis is located at the center of the gene cluster and is flanked by two regions containing genes with more common functions.  
           [0030]    The invention provides an isolated or recombinant nucleic acid encoding a capsular gene cluster of  Streptococcus  suis serotype 2, or a gene or gene fragment derived thereof, preferably as identified in FIG. 3. Genes in this gene cluster are involved in polysaccharide biosynthesis of capsular components and antigens. For a further description of such genes see, for example, Table 2. For example, a cpsA gene is provided functionally encoding regulation of capsular polysaccharide synthesis, whereas cpsB and cpsC are functionally involved in chain-in-chain length determination. Other genes, such as cpsD, E, F, G, H, I, J, K and related genes, are involved in polysaccharide synthesis, functioning, for example, as glucosyl or glycosyltransferase. The cpsF, G, H, I, J genes encode more type-specific proteins than the flanking genes which are found more-or-less conserved throughout the species and can serve as a base for selection of primers or probes in PCR-amplification or cross-hybridization experiments for subsequent cloning.  
           [0031]    The invention further provides an isolated or recombinant nucleic acid encoding a capsular gene cluster of  Streptococcus suis  serotype 1 or a gene or gene fragment derived thereof, preferably as identified in FIG. 4.  
           [0032]    In addition, the invention provides an isolated or recombinant nucleic acid encoding a capsular gene cluster of  Streptococcus suis  serotype 9 or a gene or gene fragment derived thereof, preferably as identified in FIG. 5.  
           [0033]    Furthermore, the invention provides, for example, a fragment of the  cps  locus or parts thereof, involved in the capsular polysaccharide biosynthesis, of  S. suis,  exemplified herein for serotypes 1, 2 or 9, and allows easy identification or detection of related fragments derived of other serotypes of  S. suis.    
           [0034]    The invention provides a nucleic acid probe or primer derived from a nucleic acid according to the invention allowing species or serotype specific detection of  Streptococcus suis.  Such a probe or primer (used interchangeably herein) is, for example, a DNA, RNA or PNA (peptide nucleic acid) probe hybridizing with capsular nucleic acid as provided by the invention. Species-specific detection is provided preferably by selecting a probe or primer sequence from a species-specific region (e.g. flanking region) whereas serotype-specific detection is provided preferably by selecting a probe or primer sequence from a type-specific region (e.g. central region) of a capsular gene cluster as provided by the invention. Such a probe or primer can be used in a further unmodified form, for example, in cross-hybridization or polymerase-chain reaction (PCR) experiments as, for example, described in the experimental part herein. The invention provides the isolation and molecular characterization of additional type-specific  cps  genes of  S. suis  types 1 and 9. In addition, we describe the genetic diversity of the  cps  loci of serotypes 1, 2 and 9 among the 35  S. suis  serotypes known. Type-specific probes are identified. Also, a type-specific PCR, for example, for serotype 9, is provided, being a rapid, reliable and sensitive assay used directly on nasal or tonsillar swabs or other samples of infected or carrier animals.  
           [0035]    The invention also provides a probe or primer according to the invention with at least one reporter molecule. Examples of reporter molecules are manifold and known in the art; for example, a reporter molecule can include additional nucleic acid provided with a specific sequence (e.g. oligo-dT) hybridizing to a corresponding sequence in which hybridization can easily be detected, for example, because it has been immobilized to a solid support.  
           [0036]    Yet other reporter molecules include chromophores, e.g. fluorochromes for visual detection, for example, by light microscopy or fluorescent in situ hybridization (“FISH”) techniques, or include an enzyme such as horseradish peroxidase for enzymatic detection, for example in enzyme-linked assays (“EIA”). Yet other reporter molecules include radioactive compounds for detection in radiation-based assays.  
           [0037]    In a preferred embodiment of the invention, at least one probe or primer according to the invention is provided (labeled) with a reporter molecule and a quencher molecule, together with an unlabeled probe or primer in a PCR-based test allowing rapid detection of specific hybridization.  
           [0038]    The invention further provides a diagnostic test or test kit including a probe or primer as provided by the invention. Such a test or test kit is, for example, a cross-hybridization test or PCR-based test advantageously used in rapid detection and/or serotyping of  Streptococcus suis.    
           [0039]    The invention further provides a protein or fragment thereof encoded by a nucleic acid according to the invention. Examples of such a protein or fragment are proteins described in Table 2. For example, a cpsA protein is provided that functionally encodes regulation of capsular polysaccharide synthesis, whereas cpsB and cpsC are functionally involved in chain-in-chain length determination. Other proteins or functional fragments thereof, as provided by the invention, such as cpsD, E, F, G, H, I, J, K and related proteins, are involved in polysaccharide biosynthesis, functioning, for example, as glucosyl or glycosyltransferase in polysaccharide biosynthesis of  Streptococcus suis  capsular antigen.  
           [0040]    The invention also provides a method of producing a  Streptococcus suis  capsular antigen including using a protein or functional fragment thereof as provided by the invention, and provides therewith a  Streptococcus suis  capsular antigen obtainable by such a method.  
           [0041]    A comparison of the predicted amino acid sequences of the  cps 2 genes with sequences found in the databases allowed the assignment of functions to the open reading frames. The central region contains the type-specific glycosyltransferases and the putative polysaccharide polymerase. This region is flanked by two regions encoding for proteins with common functions, such as regulation and transport of polysaccharide across the membrane. Biosynthesis of Streptococcus capsular polysaccharide antigen using a protein or functional fragment thereof is advantageously used in chemo-enzymatic synthesis and the development of vaccines which offer protection against serotype-specific Streptococcal disease, and is also advantageously used in the synthesis and development of multivalent vaccines against Streptococcal infections. Such vaccines elicit ariticapsular antibodies which confer protection.  
           [0042]    Furthermore, the invention provides an acapsular  Streptococcus  mutant for use in a vaccine, a vaccine strain derived thereof and a vaccine derived thereof. Surprisingly, and against the grain of common doctrine, the invention provides use of a  Streptococcus  mutant deficient in capsular expression in a vaccine.  
           [0043]    Acapsular  Streptococcus  mutants have long been known in the art and can be found in nature. Griffith ( J. Hyg.  27:113-159, 1928) demonstrated that pneumococci could be transformed from one type to another. If he injected live rough (acapsular or unencapsulated) type 2 pneumococci into mice, the mice would survive. If, however, he injected the same dose of live rough type 2 mixed with heat-killed smooth (encapsulated) type 1 into a mouse, the mouse would die, and, from the blood, he could isolate live smooth type 1 pneumococci. At that time, the significance of this transforming principle was not understood. However, understanding came when it was shown that DNA constituted the genetic material responsible for phenotypic changes during transformation.  
           [0044]    [0044] Streptococcus  mutants deficient in capsular expression are found in several forms. Some are fully deficient and have no capsule at all, others form a deficient capsule, characterized by a mutation in a capsular gene cluster. Deficiency can, for instance, include capsular formation wherein the organization of the capsular material has been rearranged, as, for example, demonstrable by electron microscopy. Yet others have a nearly fully developed capsule which is only deficient in a particular sugar component.  
           [0045]    Now, after much advance of biotechnology and despite the fact that little is still known about the exact localization and sequence of genes involved in capsular synthesis in Streptococci, it is possible to create mutants of Streptococci, for example, by homologous recombination or transposon mutagenesis, which has, for example, been done for GAS (Wessels et al.,  PNAS  88:8317-8321, 1991), for GBS (Wessels et al.,  PNAS  86: 8983-8987, 1989), for  S. suis  (Smith, ID-DLO Annual report 1996, page 18-19; Charland et al., Microbiol. 144:325-332, 1998) and  S. pneumoniae  (Kolkman et al., J. Bact. 178:3736-3741, 1996). Such recombinant derived mutants, or isogenic mutants, can easily be compared with the wild-type strains from which they have been derived.  
           [0046]    In a preferred embodiment, the invention provides use of a recombinant-derived  Streptococcus  mutant deficient in capsular expression in a vaccine. Recombinant techniques useful in producing such mutants are, for example, homologous recombination, transposon mutagenesis, and others, wherein deletions, insertions or (point) mutations are introduced in the genome. Advantages of using recombinant techniques include the stability of the obtained mutants (especially with homologous recombination and double crossover techniques), and the knowledge about the exact site of the deletion, mutation or insertion.  
           [0047]    In another embodiment, the invention provides a stable mutant deficient in capsular expression obtained, for example, through homologous recombination or crossover integration events. Examples of such a mutant can be found herein, for example, mutants 1OcpsB or 10cpsEF are stable mutants as provided by the invention.  
           [0048]    The invention also provides a  Streptococcus  vaccine strain and vaccine that has been derived from a  Streptococcus  mutant deficient in capsular expression. In general, the strain or vaccine is applicable within the whole range of Streptococcal infections, including animals or man or with zoonotic infections. It is, of course, now possible to first select a common vaccine strain and derive a  Streptococcus  mutant deficient in capsular expression thereof for the selection of a vaccine strain and use in a vaccine according to the invention.  
           [0049]    In a preferred embodiment, the invention provides use of a  Streptococcus  mutant deficient in capsular expression in a vaccine wherein the  Streptococcus  mutant is selected from the group composed of  Streptococcus  group A,  Streptococcus  group B,  Streptococcus suis  and  Streptococcus pneumoniae.  Herewith the invention provides vaccine strains and vaccines for use with these notoriously heterologous Streptococci, of which a multitude of serotypes exist. With a vaccine, as provided by the invention, that is derived from a specific  Streptococcus  mutant that is deficient in capsular expression, the difficulties relating to lack of heterologous protection can be circumvented since these mutants do not rely on capsular antigens, per se, to induce protection.  
           [0050]    In a preferred embodiment, the vaccine strain is selected for its ability to survive, or even replicate, in an immune-competent host or host cells and thus can persist for a certain period, varying from 1-2 days to more than one or two weeks, in a host, despite its deficient character.  
           [0051]    Although an immunodeficient host will support replication of a wide range of bacteria that are deficient in one or more virulence factors, in general, it is considered a characteristic of pathogenicity of Streptococci that they can survive for certain periods or replicate in a normal host or host cells such as macrophages. For example, Wiliams and Blakemore ( Neuropath. Appl. Neurobiol.:  16, 345-356, 1990;  Neuropath. Appl. Neurobiol.:  16, 377-392, 1990;  J. Infect. Dis.:  162, 474-481, 1990) show that both polymorphonuclear cells and macrophage cells are capable of phagocytosing pathogenic  S. suis  in pigs lacking anti- S. suis  antibodies; only pathogenic bacteria could survive and multiply inside macrophages and the pig.  
           [0052]    In a preferred embodiment, the invention, however, provides a deficient or avirulent mutant or vaccine strain which is capable of surviving at least 4-5 days, preferably at least 8-10 days in the host, thereby allowing the development of a solid immune response to subsequent  Streptococcus  infection.  
           [0053]    Due to its persistent but avirulent character, a  Streptococcus  mutant or vaccine strain, as provided by the invention, is well suited to generate specific and/or long-lasting immune responses against Streptococcal antigens. Moreover, possible specific immune responses of the host directed against a capsule are relatively irrelevant because a vaccine strain, as provided by the invention, is typically not recognized by such antibodies.  
           [0054]    In addition, the invention provides a  Streptococcus  vaccine strain according the invention, which strain includes a mutant capable of expressing a  Streptococcus  virulence factor or antigenic determinant.  
           [0055]    In a preferred embodiment, the invention provides a  Streptococcus  vaccine strain, according to the invention, which includes a mutant capable of expressing a  Streptococcus  virulence factor wherein the virulence factor or antigenic determinant is selected from a group of cellular components, such as muramidase-released protein (“MRP”), extracellular factor (“EF”) and cell-membrane associated proteins 60kDA heat shock protein, pneumococcal surface protein A (Psp A), pneumolysin, C protein, protein M, fimbriae, hemagglutinins and hemolysin or components functionally related thereto.  
           [0056]    In a preferred embodiment, the invention provides a  Streptococcus  vaccine strain including a mutant capable of over-expressing the virulence factor. In this way, the invention provides a vaccine strain for incorporation in a vaccine which specifically causes a host immune response directed against antigenically important determinants of virulence (listed above), thereby providing specific protection against the determinants. Over-expression can, for example, be achieved by cloning the gene involved behind a strong promoter, which is, for example, constitutionally expressed in a multicopy system, either in a plasmid or via intergration in a genome.  
           [0057]    In yet another embodiment, the invention provides a  Streptococcus  vaccine strain, according to the invention, including a mutant capable of expressing a non- Streptococcus  protein. Such a vector- Streptococcus  vaccine strain allows, when used in a vaccine, protection against pathogens other than  Streptococcus.    
           [0058]    Due to its persistent but avirulent character, a  Streptococcus  vaccine strain or mutant as provided by the invention is well suited to generate specific and long-lasting immune responses, not only against Streptococcal antigens, but also against other antigens expressed by the strain. Specifically, antigens derived from another pathogen are now expressed without the detrimental effects of the antigen or pathogen which would otherwise have harmed the host.  
           [0059]    An example of such a vector is a  Streptococcus  vaccine strain or mutant wherein the antigen is derived from a pathogen, such as  Actinobacilluspleuropneumonia, Mycoplasmatae, Bordetella, Pasteurella, E. coli, Salmonella, Campylobacter, Serpulina  and others.  
           [0060]    The invention also provides a vaccine including a  Streptococcus  vaccine strain or mutant according to the invention and a pharmaceutically acceptable carrier or adjuvant. Carriers or adjuvants are well known in the art; examples are phosphate buffered saline, physiological salt solutions, (double-) oil-in-water emulsions, aluminumhydroxide, Specol, block- or co-polymers, and others.  
           [0061]    A vaccine according to the invention can include a vaccine strain either in a killed or live form. For example, a killed vaccine including a strain having (over) expressed a Streptococcal or heterologous antigen or virulence factor is very well suited for eliciting an immune response. In a preferred embodiment, the invention provides a vaccine wherein the strain is live, due to its persistent but avirulent character; a  Streptococcus  vaccine strain, as provided by the invention, is well suited to generate specific and long-lasting immune responses.  
           [0062]    The invention also provides a method for controlling or eradicating a Streptococcal disease in a population comprising vaccinating subjects in the population with a vaccine according to the invention.  
           [0063]    In a preferred embodiment, a method for controlling or eradicating a Streptococcal disease is provided including testing a sample, such as a blood sample, or nasal or throat swab, feces, urine, or other samples such as can be sampled at or after slaughter, collected from at least one subject, such as an infant or a pig, in a population partly or wholly vaccinated with a vaccine according to the invention for the presence of encapsulated Streptococcal strains or mutants. Since a vaccine strain or mutant according to the invention is not pathogenic, and can be distinguished from wild-type strains by capsular expression, the detection of (fully) encapsulated Streptococcal strains indicates that wild-type infections are still present. Such wild-type infected subjects can then be isolated from the remainder of the population until the infection has passed. With domestic animals, such as pigs, it is even possible to remove the infected subject from the population as a whole by culling. Detection of wild-type strains can be achieved via traditional culturing techniques, or by rapid detection techniques such as PCR detection.  
           [0064]    In yet another embodiment, the invention provides a method for controlling or eradicating a Streptococcal disease including testing a sample collected from at least one subject in a population partly or wholly vaccinated with a vaccine according to the invention for the presence of capsule-specific antibodies directed against Streptococcal strains. Capsule specific antibodies can be detected with classical techniques known in the art, such as used for Lancefield&#39;s group typing or serotyping.  
           [0065]    A preferred embodiment for controlling or eradicating a Streptococcal disease in a population includes vaccinating subjects in the population with a vaccine according to the invention and testing a sample collected from at least one subject in the population for the presence of encapsulated Streptococcal strains and/or for the presence of capsule-specific antibodies directed against Streptococcal strains.  
           [0066]    For example, a method is provided wherein the Streptococcal disease is caused by  Streptococcus suis.    
           [0067]    The invention also provides a diagnostic assay for testing a sample for use in a method according to the invention including at least one means for the detection of encapsulated Streptococcal strains and/or for the detection of capsule-specific antibodies directed against Streptococcal strains.  
           [0068]    The invention further provides a vaccine including an antigen according to the invention and a suitable carrier or adjuvant. The immunogenicity of a capsular antigen provided by the invention is, for example, increased by linking to a carrier (such as a carrier protein), allowing the recruitment of T-cell help in developing an immune response.  
           [0069]    The invention further provides a recombinant microorganism provided with at least a part of a capsular gene cluster derived from  Streptococcus suis.  The invention provides, for example, a lactic acid bacterium provided with at least a part of a capsular gene cluster derived from  Streptococcus suis.  Various food-grade lactic acid bacteria ( Lactococcus lactis, Lactobacillus casei, Lactobacillus plantarium  and  Streptococcus gordonii ) have been used as delivery systems for mucosal immunization. It has now been shown that oral (or mucosal) administration of recombinant  L. lactis, Lactobacillus,  and  Streptococcus gordonii  can elicit local IgA and/or IgG antibody responses to an expressed antigen. The use of oral routes for immunization against infective diseases is desirable because oral vaccines are easier to administer and have higher compliance rates, and because mucosal surfaces are the portals of entry for many pathogenic microbial agents. It is within the skill of the artisan to provide such micro-organisms with (additional) genes.  
           [0070]    The invention further provides a recombinant  Streptococcus suis  mutant provided with a modified capsular gene cluster. It is within the skill of the artisan to swap genes within a Species. In a preferred embodiment, an avirulent  Streptococcus suis  mutant is selected to be provided with at least a part of a modified capsular gene cluster according to the invention.  
           [0071]    The invention further provides a vaccine including a microorganism or a mutant provided by the invention. An advantage of such a vaccine over currently used vaccines is that they include accurately defined microorganisms and well-characterized antigens, allowing accurate determination of immune responses against various antigens of choice. 
       
    
    
       [0072]    The invention is further explained in the experimental part of this description without limiting the invention thereto.  
       DESCRIPTION OF THE FIGURES  
       [0073]    [0073]FIG. 1 illustrates the organization of the cps2 gene cluster of  S. suis  type  2 .  
         [0074]    (A) Genetic map of the cps2 gene cluster. The shadowed arrows represent potential ORFs. Interrupted ORFs indicate the presence of stop codons or frame-shift mutations. Gene designations are indicated below the ORFs. The closed arrows indicate the position of the potential promoter sequences. I indicates the position of the potential transcription regulator sequence. III indicates the position of the 100-bp repeated sequence.  
         [0075]    (B) Physical map of the cps2 locus. Restriction sites are as follows: A:  Alul;  C:  ClaI,  E:  EcoRI;  H:  HindIII;  K:  KpnI ; M:  MluI;  N:  NsiI;  P:  PstI;  S:  SnaBI;  Sa:  SacI;  X:  XbaI.    
         [0076]    (C) The DNA fragments cloned in the various plasmids.  
         [0077]    [0077]FIG. 2 illustrates ethidium bromide stained agarose gel showing PCR products obtained with chromosomal DNA of  S. suis  strains belonging to the serotypes 1,2, ½, 2, 9 and 14 and  cps 2 J, cpsII,  and  cps 9 H  primer sets as described herein.  
         [0078]    (A)  cpsII  primers; (B)  cps 2 J  primers and (C)  cps 9 H  primers.  
         [0079]    Lanes 1-3: serotype 1 strains; lanes 4-6: serotype 2 strains; lanes 7-9: serotype ½ strains; lanes 10-12: serotype 9 strains and lanes 13-15: serotype 14 strains.  
         [0080]    (B) Ethidium bromide stained agarose gel showing PCR products obtained with tonsillar swabs collected from pigs carrying  S. suis  type  2 , type  1  or type  9  strains and  cps 2 J, cpsII  and  cpsH  primer sets as described in Materials and Methods. Bacterial DNA suitable for PCR was prepared by using the multiscreen methods as described previously ( 20 ).  
         [0081]    (C)  cpsII  primers. (B)  cps 2 J  primers and (C)  cps 9 H  primers.  
         [0082]    Lanes 1-3: PCR products obtained with tonsillar swabs collected from pigs carrying  S. suis  type 1 strains; lanes 4-6: PCR products obtained with tonsillar swabs collected from pigs carrying  S. suis  type 2 strains; lanes 7-9: PCR products obtained with tonsillar swabs collected from pigs carrying  S. suis  type 9 strains; lanes 10-12: PCR products obtained with chromosomal DNA from serotype 9, 2 and 1 strains respectively; lane 13: negative control, no DNA present.  
         [0083]    [0083]FIG. 3 illustrates the CPS2 nucleotide sequences and corresponding amino acid sequences from the open reading frames.  
         [0084]    [0084]FIG. 4 illustrates the CPS 1 nucleotide sequences and corresponding amino acid sequences from the open reading frames.  
         [0085]    [0085]FIG. 5 illustrates the CPS9 nucleotide sequences and corresponding amino acid sequences from the open reading frames.  
         [0086]    [0086]FIG. 6 illustrates the CPS7 nucleotide sequences and corresponding amino acid sequences from the open reading frames.  
         [0087]    [0087]FIG. 7 illustrates alignment of the N-terminal parts of Cps2J and Cps2K.  
         [0088]    Identical amino acids are marked by bars. The amino acids shown in bold are also conserved in CPS14I Cpsl4J of  S. pneumoniae  and several other glycosyltransferases ( 19 ). The aspartate residues marked by asterisks are strongly conserved.  
         [0089]    [0089]FIG. 8 illustrates transmission electron micrographs of thin sections of various  S. suis  strains.  
         [0090]    (A) wild type strain  10 ;  
         [0091]    (B) mutant strain 10cpsB;  
         [0092]    (C) mutant strain 10cpsEF.  
         [0093]    Bar=100 nm  
         [0094]    [0094]FIG. 9 illustrates the kinetics of phagocytosis of wild type and mutant  S. suis  strains.  
         [0095]    (A) Kinetics of phagocytosis of wild type and mutant  S. suis  strains by porcine alveolar macrophages. Phagocytosis was determined as described herein. The Y-axis represents the number of CFU per milliliter in the supernatant fluids as determined by plate counting, the X-axis represents time in minutes.  
         [0096]    □ wild type strain  10 ;  
         [0097]    ο mutant strain 10cpsB;  
         [0098]    Δ mutant strain 10cpsEF.  
         [0099]    (B) Kinetics of intracellular killing of wild type and mutant  S. suis  strains by porcine AM. The intracellular killing was determined as described herein. The Y-axis represents the number of CFU per ml in the supernatant fluids after lysis of the macrophages as determined by plate counting, the X-axis represents time in minutes.  
         [0100]    □ wild type strain  10 ;  
         [0101]    ο mutant strain 10cpsB;  
         [0102]    Δ mutant strain 10cpsEF.  
         [0103]    [0103]FIG. 10 illustrates the nucleotide sequence alignment of the highly conserved 100-bp repeated element.  
         [0104]    (1) 100-bp repeat between cps2G and cps2H  
         [0105]    (2) 100-bp repeat within “cps2M”  
         [0106]    (3) 100-bp repeat between cps2O and cps2P  
         [0107]    [0107]FIG. 11 illustrates the cps2, cps9 and cps7 gene clusters of  S. suis  serotypes  2 ,  9  and  7 .  
         [0108]    (A) Genetic organization of the cps2 gene cluster [84]. The large arrows represent potential ORFs. Gene designations are indicated below the ORFs. Identically filled arrows represent ORFs which showed homology. The small closed arrows indicate the position of the potential promoter sequences. | indicates the position of the potential transcription regulator sequence.  
         [0109]    (B) Physical map and genetic organization of the cps9 gene cluster [15]. Restriction sites are as follows: B:  BamHI;  P:  PstI;  H:  HindIII;  X:  XbaI.  The DNA fragments cloned in the various plasmids are indicated. The open arrows represent potential ORFs.  
         [0110]    (C) Physical map and genetic organization of the cps7 gene cluster. Restriction sites are as follows: C:  Clal;  P:  PstI;  Sc:  ScaI.  The DNA fragments cloned in the various plasmids are indicated. The open arrows represent potential ORFs.  
         [0111]    [0111]FIG. 12 illustrates ethidium bromide stained agarose gel showing PCR products.  
         [0112]    (A) Ethidium bromide stained agarose gel showing PCR products obtained with chromosomal DNA of  S. suis  strains belonging to the serotypes 1, 2, 9 and 7 and the cps7H primer set. Strain designations are indicated above the lanes. C: negative control, no DNA present. M: molecular size marker (lambda digested with  EcoRI  and  HindIII ).  
         [0113]    (B) Ethidium bromide stained agarose gel showing PCR products obtained with serotype 7 strains collected in different countries and from different organs. Bacterial DNA suitable for PCR was prepared by using the multiscreen method as described herein [89]. Strain designations are indicated above the lanes. M: molecular size marker (lambda digested with  EcoRI  and HindIII).  
     
    
     DETAILED DESCRIPTION OF THE INVENTION  
       [0114]    Experimental part  
         [0115]    MATERIAL AND METHODS  
         [0116]    Bacterial strains and growth conditions.  
         [0117]    The bacterial strains and plasmids used in this study are listed in Table 1.  S. suis  strains were grown in Todd-Hewitt broth (code CM189, Oxoid), and plated on Columbia agar blood base (code CM33 1, Oxoid) containing 6% (v/v) horse blood.  E. coli  strains were grown in Luria broth (28) and plated on Luria broth containing 1.5% (w/v) agar. If required, antibiotics were added to the plates at the following concentrations: spectinomycin: 100 μg/mi for  S. suis  and 50 μg/ml for  E. coli  and ampicillin, 50 μg/nl.  
         [0118]    Serotyping. The  S. suis  Strains were serotyped by the slide agglutination test with serotype-specific antibodies (44).  
         [0119]    DNA techniques. Routine DNA manipulations were performed as described by Sambrook et al. (36).  
         [0120]    Alkaline phosphatase activity. To screen for PhoA fusions in  E. coli,  plasmid libraries were constructed. Therefore, chromosomal DNA of  S. suis  type 2 was digested with  AIuI.  The 300-500-bp fragments were ligated to Smal-digested pPHOS2. Ligation mixtures were transformed to the PhoA  E. coli  strain CC118. Transformants were plated on LB media supplemented with 5-Bromo-4-chloro-3-indolylfosfaat (BCIP, 50 μg/ml, Boehringer, Mannheim, Germany). Blue colonies were purified on fresh LB/BCIP plates to verify the blue phenotype.  
         [0121]    DNA sequence analysis. DNA sequences were determined on a 373A DNA Sequencing System (Applied Biosystems, Warrington, GB)- Samples were prepared by using an ABI/PRISM dye terminator cycle sequencing ready reaction kit (Applied Biosystems). Sequencing data were assembled and analyzed using the MacMollyTetra program. Custom-made sequencing primers were purchased from Life Technologies. Hydrophobic stretches within proteins were predicted by the method of Klein et al. ( 17 ). The BLAST program available on Netscape Navigator™ was used to search for protein sequences related to the deduced amino acid sequences.  
         [0122]    Construction of gene-specific knock-out mutants of  S. suis.  To construct the mutant strains 10cpsB and 10cpsEF, we electrotransformed the pathogenic serotype 2 strain 10 (45, 49) of  S. suis  with pCPS 11 and pCPS28 respectively. In these plasmids, the cpsB and cpsEF genes were disturbed by the insertion of a spectinomycin-resistance gene. To create pCPS 11, the internal 400 bp  PstIBamHI  fragment of the  cpsB  gene in pCPS7 was replaced by the Spc R  gene. For this purpose, pCPS7 was digested with  PstI  and BamHI and ligated to the 1,200-bp PstI- BamHi  fragment, containing the Spc R  gene, from pIC-spc. To construct pCPS28, we have used pIC20R. In this plasmid we inserted the KpnI-SalI fragment from pCPS17 (resulting in pCPS25) and the  XbaI - ClaI  fragment from pCPS20 (resulting in pCPS27). pCPS27 was digested with  PstI  and  XhoI  and ligated to the 1,200-bp  Pstl - Xhol  fragment, containing the Spc R  gene of pIC-spc. The electrotransformation to  S. suis  was carried out as described before (38).  
         [0123]    Southern blotting and hybridization. Chromosomal DNA was isolated as described by Sambrook et al. (36). DNA fragments were separated on 0.8% agarose gels and transferred to Zeta-Probe GT membranes (Bio-Rad) as described by Sambrook et al. (36). DNA probes were labeled with [( −32 p] dCTP (3000 Ci mmol −1 ; Amersham) by use of a random primed labeling kit (Boehringer). The DNA on the blots was hybridized at 65° C. with appropriate DNA probes as recommended by the supplier of the Zeta-Probe membranes. After hybridization, the membranes were washed twice with a solution of 40 mM sodium phosphate, pH 7.2, 1 mM EDTA, 5% SDS for 30 min at 65° C. and twice with a solution of 40 mM sodium phosphate, pH 7.2, 1 mM EDTA, 1% SDS for 30 min at 65° C.  
         [0124]    PCR. The primers used in the  cps 2J PCR correspond to the positions 13791-13813 and 14465-14443 in the  S. suis  cps2 locus. The sequences were: 5′-CAAACGCAAGGAATTACGGTATC-3′ (SEQ. ID. No. 1) and 5′-GAGTATCTAAAGAATGCCTATTG-3′ (SEQ. ID. No. 2). The primers used for the  cpslI  PCR correspond to the positions 4398-4417 and 4839-4821 in the  S. suis cpsI  sequence. The sequences were: 5′-GGCGGTCTAGCAGATGCTCG-3′ (SEQ. ID. No. 3) and 5′-GCGAACTGTTAGCAATGAC-3′ (SEQ. ID. No. 4). The primers used in the  cps 9H PCR correspond to the positions 4406-4126 and 4494-4475 in the  S. suis cps 9 sequence. The sequences were: 5′-GGCTACATATAATGGAAGCCC3′ (SEQ. ID No. 5) and 5′-CGGAAGTATCTGGGCTACTG-3′ (SEQ. ID. No. 6).  
         [0125]    Construction of gene-specific knock-out mutants of  S. suis.  To construct the mutant strains 10cpsB and 10cpsEF, we electrotransformed the pathogenic serotype 2 strain 10 of  S. suis  with pCPS 11 and pCPS28 respectively. In these plasmids, the  cpsB  and cpsEF genes were disturbed by the insertion of a spectinomycin-resistance gene. To create pCPS11, the internal 400 bp  PstI - BamHI  fragment of the  cpsB  gene in pCPS7 was replaced by the Spc R  gene. For this purpose, pCPS7 was digested with  Psti  and BamHI and ligated to the 1,200-bp  PstI - BamHI  fragment, containing the Spc R  gene, from pIC-spc. To construct pCPS28, we have used pIC20R. In this plasmid, we inserted the  KpnI - SalI  fragment from pCPS17 (resulting in pCPS25) and the  XbaI - ClaI  fragment from pCPS20 (resulting in pCPS27). pCPS27 was digested with  PsI  and  XhoI  and ligated to the 1,200-bp  PstI - XhoI  fragment, containing the Spc R  gene of pIC-spc. The electrotransformation to  S. suis  was carried out as described before (38).  
         [0126]    Phagocytosis assay. Phagocytosis assays were performed as described by Leij et al. (23). Briefly, to opsonize the cells, 10 7    S. suis  cells were incubated with 6% SPF-pig serum for 30 min at 37° C. in a head-over-head rotor at 6 rpm. 10 7  AM and 10 7  opsonized  S. suis  cells were combined and incubated at 37° C. under continuous rotation at 6 rpm. At 0, 30, 60 and 90 min, 1- ml samples were collected and mixed with 4 ml of ice-cold EMEM to stop phagocytosis. Phagocytes were removed by centrifugation for 4 min at 110×g and 4° C. The number of colony-forming units, (“CFU”) in the supernatants was determined. Control experiments were carried out simultaneously by combining 10 7  opsonized  S. suis  cells with EMEM (without AM).  
         [0127]    Killing assays. AM (10 7 /ml) and opsonized  S. suis  cells (10 7 /ml) were mixed 1:1 and incubated for 10 min at 37° C. under continuous rotation at 6 rpm. Ice-cold EMEM was added to stop further phagocytosis and killing. To remove extracellular  S. suis  cells, phagocytes were washed twice (4 min, 110×g, 4° C.) and resuspended in 5 ml EMEM containing 6% SPF serum. The tubes were incubated at 37° C. under rotation at 6 rpm. After 0, 15, 30, 60 and 90 min, samples were collected and mixed with ice-cold EMEM to stop further killing. The samples were centrifuged for 4 min at 110×g at 4° C. and the phagocytic cells were lysed in EMEM containing 1% saponine for 20 min at room temperature. The number of CFU in the suspensions was determined.  
         [0128]    Pigs. Germfree pigs, crossbreeds of Great Yorkshire and Dutch Landrace, were obtained from sows by caesarian sections. The surgery was performed in sterile flexible film isolators. Pigs were allotted to groups, each consisting of 4 pigs, and were housed in sterile stainless steel incubators.  
         [0129]    Experimental infections. Pigs were inoculated intranasally with  S. suis  type 2 as described before. To predispose the pigs for infection with  S. suis,  five-day old pigs were inoculated intranasally with about 10 7  CFU of  Bordetella bronchiseptica  strain 92932. Two days later, the pigs were inoculated intranasally with  S. suis  type 2 (106 CFU). Pigs were monitored twice daily for clinical signs of disease, such as fever, nervous signs and lameness. Blood samples were collected three times a week from each pig. White blood cells were counted with a cell counter. To monitor infection with  S. suis  and  B. bronchiseptica  and to check for absence of contaminants, we collected swabs of nasopharynx and feces daily. The swabs were plated directly onto Columbia agar containing 6% horse blood. After three weeks, the pigs were killed and examined for pathological changes. Tissue specimens from the central nervous system, serosae, and joints were examined bacteriologically and histologically as described herein (45, 49). Colonization of the serosae was scored positively when  S. suis  was isolated from the pericardium, thoracal pleura or the peritoneum. Colonization of the joints was scored positively when  S. suis  was isolated from one or more joints (12 joints per animal were scored).  
         [0130]    Vaccination and challenge. One week old pigs were vaccinated intravenously with a dosage of 106 cfu of the  S. suis  strains 10cpsEF or 10cpsB. Three weeks later, the pigs were challenged intravenously with the pathogenic Serotype 2 strain 10 (107 cfu). Disease monitoring, hematological, serological and bacteriological examinations as well as post-mortum examinations were as described before under experimental infections.  
         [0131]    Electron Microscopy. Bacteria were prepared for electron microscopy as described by Wagenaar et al. (50). Shortly, bacteria were mixed with agarose ND (Boehringer) of 37° C. to a concentration of 0.7%. The mixture was immediately cooled on ice. Upon gelifying, samples were cut into 1 to 1.5 mm slices and incubated in a fixative containing 0.8% glutaraldehyde and 0.8% osmiumtetraoxide. Subsequently, the samples were fixed and stained with uranyl acetate by microwave stimulation, dehydrated and imbedded in eponaraldite resin. Ultra-thin sections were counterstained with lead citrate and examined with a Philips CM 10 electron microscope at 80 kV (FIG. 8).  
         [0132]    Isolation of porcine alveolar macrophages (AM). Porcine AM were obtained from the lungs of specific pathogen free (“SPF”) pigs. Lung lavage samples were collected as described by van Leengoed et al. (43). Cells were suspended. in EMEM containing 6% (v/v). SPF-pig serum and adjusted to 10 7  cells per ml.  
       RESULTS  
       [0133]    Identification of the cps locus.  
         [0134]    The cps locus of  S. suis  type 2 was identified through a strategy developed for the genetic identification of exported proteins (13, 31). In this system, we used a plasmid (pPHOS2) containing a truncated alkaline phosphatase gene (13). The gene lacked the promoter sequence, the translational start site and the signal sequence. The truncated gene is preceded by a unique Smal restriction site. Chromosomal DNA of  S. suis  type 2, digested with  AluI,  was randomly cloned in this restriction site. Because translocation of PhoA across the cytoplasmic membrane of  E. coli  is required for enzymatic activity, the system can be used to select for  S. suis  fragments containing a promoter sequence, a translational start site and a functional signal sequence. Among 560 individual  E. coli  clones tested, 16 displayed a dark blue phenotype when plated on media containing BCIP. DNA sequence analysis of the inserts from several of these plasmids was performed (results not shown) and the deduced amino acid sequences were analyzed. The hydrophobicity profile of one of the clones (pPHOS7, results not shown) showed that the N-terminal part of the sequence resembled the characteristics of a typical signal peptide: a short hydrophilic N-terminal region is followed by a hydrophobic region of 38 amino acids. These data indicate that the phoA system was successfully used for the selection of  S. suis  genes encoding exported proteins. Moreover, the sequences were analyzed for similarities present in the databases. The sequence of pPHOS7 showed a high similarity (37% identity) with the protein encoded by the  cps 14 C  gene of  Streptococcus pneumoniae  (19). This strongly suggests that pPHOS7 contains a part of the  cps  operon of  S. suis  type 2.  
         [0135]    Cloning of the flanking  cps  genes. In order to clone the flanking  cps  genes of  S. suis  type 2, the insert of pPHOS7 was used as a probe to identify chromosomal DNA fragments which contain flanking  cps  genes. A 6-kb  HindIII  fragment was identified and cloned in pKUN19. This yielded clone pCPS6 (FIG. 1, part C). Sequence analysis of the insert of pCPS6 revealed that pCPS6 most probably contained the 5′-end of the cps locus, but still lacked the 3′-end. Therefore, sequences of the 3′-end of pCPS6 were in turn used as a probe to identify chromosomal fragments containing  cps  sequences located further downstream. These fragments were also cloned in pKUN19, resulting in pCPS17. Using the same system of chromosomal walking, we subsequently generated the plasmids pCPS18, pCPS20, pCPS23 and pCPS26, containing downstream  cps  sequences.  
         [0136]    Analysis of the cps operon. The complete nucleotide sequence of the cloned fragments was determined (FIG. 4). Examination of the compiled sequence revealed the presence of at least 13 potential open reading frames (Orfs), which were designated as Orf 2Y, Orf2X and Cps2A-Cps2K (FIG. 1, part A, FIG. 1, part A). Moreover, a 14th, incomplete Orf (Orf 2Z) was located at the 5′-end of the sequence. Two potential promoter sequences were identified. One was located 313 bp (locations 1885-1865 and 1884-1889) upstream of Orf2X. The other potential promoter sequence was located 68 bp upstream of Orf2Y (locations 2241-2236 and 2216-2211). Orf2Y is expressed in opposite orientation. Between Orfs 2Y and 2Z, the sequence contained a potential stem-loop structure, which could act as a transcription terminator. Each Orf is preceded by a ribosome-binding site and the majority of the Orfs are very closely linked. The only significant intergenic gap was found between Cps2G and Cps2H (389 nucleotides). However, no obvious promoter sequences or potential stem-loop structures were found in this region. These data suggest that Orf2X and Cps2A-Cps2K are arranged as an operon.  
         [0137]    An overview of all Orfs with their properties is shown in Table 2. The majority of the predicted gene products is related to proteins involved in polysaccharide biosynthesis. Orf2Z showed some similarity with the YitS protein of  Bacillus subtilis.  YitS was identified during the sequence analysis of the complete genome of  B. subtilis.  The function of the protein is unknown.  
         [0138]    Orf2Y showed similarity with the YcxD protein of  B. subtilis  (53). Based on the similarity between YcxD and MocR of  Rhizohium meliloti  (33), YcxD was suggested to be a regulatory protein.  
         [0139]    Orf2X showed similarity with the hypothetical YAAA proteins of  Haemophilus influenzae  and  E. coli.  The function of these proteins is unknown.  
         [0140]    The gene products encoded by the  cps 2 A, cps 2 B, cps 2 C  and  cps 2 D  genes showed approximate similarity with the CpsA, CpsC, CpsD and CpsB proteins of several serotypes of  Streptococcus pneumoniae  (19), respectively. This suggests similar functions for these proteins. Hence, Cps2A may have a role in the regulation of the capsular polysaccharide synthesis. Cps2B and Cps2C could be involved in the chain length determination of the type 2 capsule and Cps2C can play an additional role in the export of the polysaccharide. The Cps2D protein of  S. suis  is related to the CpsB protein of  S. pneumoniae  and to proteins encoded by genes of several other Gram-positive bacteria involved in polysaccharide or exopolysaccharide synthesis, but their function is unknown (19).  
         [0141]    The protein encoded by the  cps 2 E  gene showed similarity to several bacterial proteins with glycosyltransferase activities Cps14E and Cps19fE of  S. pneumoniae  serotypes 14 and 19F (18, 19, 29), CpsE of  Streptococcus salvarius  (X94980) and CpsD of  Streptococcus agalactiae  (34). Recently, Kolkman et al. (18) showed that Cps14E is a glucosyl-1-phosphate transferase that links glucose to a lipid carrier, the first step in the biosynthesis of the  S. pneumoniae  type 14 repeating unit. Based on these data, a similar function may be fulfilled by Cps2E of  S. suis.    
         [0142]    The protein encoded by the  cps 2 F  gene showed similarity to the protein encoded by the  rfbU  gene of  Salmonella enteritica.  (25). This similarity is most pronounced in the C-terminal regions of these proteins. The  rfbU  gene was shown to encode mannosyltransferase activity (25).  
         [0143]    The  cps 2 G  gene encoded a protein that showed moderate similarity with the  rfbF  gene product of  Campylohacter hyoilei  (22), the  epsF  gene product of  S. thermophilus  (40) and the  capM  gene product of  S. aureus  (24). On the basis of similarity, the  rfbF, epsF  and  capM  genes are suggested to encode galactosyltransferase activities. Hence, a similar glycosyltransferase activity could be fulfilled by the  cps 2 G  gene product.  
         [0144]    The  cps 2 H  gene encodes a protein that is similar to the N-terminal region of the  IgtI ) gene product of  Haemophilus influenzae  (U32768). Moreover, the hydrophobicity plots of Cps2H and LgtD looked very similar in these regions (data not shown). Based on sequence similarity, the  IgtD  gene product was suggested to have glycosyltransferase activity (U32768).  
         [0145]    The gene product encoded by the  cps 2 I  gene showed some similarity with a protein of  Actinobacillus actinomycetemcomitans  (AB002668). This protein is part of the gene cluster responsible for the serotype-b-specific antigen of  A. actinomycetemcomitans.  The function of the protein is unknown.  
         [0146]    The gene products encoded by the  cps 2 J  and  cps 2 K  genes showed significant similarities to the Cps14J protein of  S. pneumoniae.  The cps14J gene of  S. pneumoniae  was shown to encode a β-1,4-galactosyltransferase activity. In  S. pneumoniae,  CpsJ is responsible for the addition of the fourth (i.e. last) sugar in the synthesis of the  S. pneumoniae  serotype 14 polysaccharide (20). Even some similarity was found between Cps2J and Cps2K (FIG. 2, 25.5% similarity). This similarity was most pronounced in the N-terminal regions of the proteins (FIG. 7). Recently, two small conserved regions were identified in the N-terminus of Cpsl4J and Cpsl4I and their homologues (20). These regions were predicted to be important for catalytic activity. Both regions, DXS and DXDD (FIG. 2), were also found in Cps2J and Cps2K.  
         [0147]    Distribution of the  cps 2 genes in other  S. suiss  serotypes. To examine the relationship between the  cps 2 genes and cps genes in the other  S. suis  serotypes, we performed crosshybridization experiments. DNA fragments of the individual  cps 2 genes were amplified by PCR, labeled with  32 P, and used to probe Southern blots of chromosomal DNA of the reference strains of the 35 different  S. suis  serotypes. Large variations in the hybridization patterns were observed (Table 4). As a positive control, we used a probe specific for 16S rRNA. The 16S rRNA probe hybridized with all serotypes tested. However, none of the other genes tested were common in all serotypes. Based on the genetic organization of the genes, we previously suggested that  orfX  and  cpsA - cpsK  genes are part of one operon and that the proteins encoded by these genes are all involved in polysaccharide biosynthesis. OrfY and OrfZ are not a part of this operon, and their role in the polysaccharide biosynthesis is unclear. Based on sequence similarity data,  OrfY  may be involved in regulation of the  cps 2 genes. OrfZ is proposed to be unrelated to polysaccharide biosynthesis. Probes specific for the  orfZ, orfY, orfX, cpsA, cpsB, cpsC  and  cpsD  genes hybridized with most other serotypes. This suggests that the proteins encoded by these genes are not type-specific, but may perform more common functions in biosynthesis of the capsular polysaccharide. This confirms previous data which showed that the  cps 2 A - cps 2 D  genes showed strong similarity to  cps  genes of several serotypes of  Streptococcus pneumoniae.  Based on this similarity, Cps2A is possibly a regulatory protein, whereas Cps2B and Cps2C may play a role in length determination and export of polysaccharide. The  cps 2 E  gene hybridized with DNA of Serotypes 1, 2, 14 and ½. The  cps 2 E  gene showed a strong similarity to the  cps 14 E  gene of  S. pneumoniae  (18). This enzyme was shown to have a glucosyl-1-phosphate activity and catalyzed the transfer of glucose to a lipid carrier (18). These data indicate that a glycosyltransferase closely related to Cps14E may be responsible for the first step in the biosynthesis of polysaccharide in the  S. suis  serotypes 1, 2, 14 and ½. The  cps 2 F, cps 2 G, cps 2 H, cps 2 I  and  cps 2 J  genes hybridized with chromosomal DNA of serotypes 2 and ½ only. The  cps 2 G  gene showed an additional weak hybridization signal with DNA of serotype 34. In agglutination tests, serotype ½ showed agglutination with sera specific for serotype 2 as well as with sera specific for serotype 1. This suggests that serotype ½ shares antigenic determinants with both types 1 and 2. The hybridization data confirmed these data. All putative glycosyltransferases present in serotype 2 are also present in serotype ½. The  cps 2 K  gene showed a hybridization pattern similar to the  cps 2 E  gene. Hybridization was observed with DNA of serotypes 1, 2, 14 and ½. Taken together, these hybridization data show that the  cps 2 gene cluster can be divided into three regions: a central region containing the type-specific genes is flanked by two regions containing common genes for various serotypes.  
         [0148]    Cloning of the type-specific cps genes of serotypes 1 and 9. To clone the type-specific  cps  genes of  S. suis  serotype 1, we used the  cps 2 E  gene as a probe to identify chromosomal DNA fragments of type 1 which contain flanking  cps  genes. A 5 kb  EcoRV  fragment was identified and cloned in pKUN19. This yielded pCPS1-1 (FIG. 1, part B). This fragment was in turn used as a probe to identify an overlapping 2.2 kb HindIII fragment. pKUN19 containing this HindIII fragment was designated pCPS1-2. The same strategy was followed to identify and clone the type-specific cps genes of serotype 9. In this case, we used the  cps 2 D  gene as a probe. A 0.8 kb HindIII-XbaI fragment was identified and cloned, yielding pCPS9-1 (FIG. 1, part C). This fragment was in turn used as a probe to identify a 4 kb XbaI fragment. pKUN19 containing this 4 kb XbaI fragment was designated pCPS9-2.  
         [0149]    Analysis of the cloned  cps 1 genes. The complete nucleotide sequence of the inserts of pCPSI-1 and pCPS1-2 was determined (FIG. 5). Examination of the sequence revealed the presence of five complete and two incomplete Orfs (FIG. 1, part B). Each Orf is preceded by a ribosome-binding site. In accord with data obtained for the  cps 2 genes of serotype 2, the majority of the Orfs is very closely linked. The only significant gap (718 bp) was found between Cps1G and Cpsp1H. No obvious promoter sequences or potential stem-loop structures could be found in this region. This suggests that, as in serotype 2, the  cps  genes in serotype 1 are arranged in an operon.  
         [0150]    An overview of the Orfs and their properties is shown in Table 2. As expected on the basis of the hybridization data (Table 4), the protein encoded by the  cps 1 E  gene was related to Cps2E of  S. suis  type 2 (identity of 86%). The fragment cloned in pCPS1-1 lacked the coding region for the first 7 amino acids of the  cps 1 E  gene.  
         [0151]    The protein encoded by the  cps 1 F  and  cps 1 G  genes showed strong similarity to the Cps14F and Cps 1 14G proteins of  Streptococcus pneumoniae  serotype 14, respectively (20). The function of the Cps14F is not completely clear, but it has been suggested that Cps14F has a role in glycosyltransferase activity. The  cps 14G gene of  S. pneumoniae  was shown to encode β-1, 4-galactosyltransferase activity. In  S. pneumoniae  type 14, this activity is required for the second step in the biosynthesis of the oligosaccharide subunit (20). Based on the similarity of the data, similar glyco syltransferase and enhancing activities are suggested for the cps1G and  cps 1 F  genes of  S. suis  type 1.  
         [0152]    The protein encoded by the  cps 1 H  gene showed similarity to the Cps14M protein of  S. pneumoniae  (20). Based on sequence similarity, Cps14H was proposed to be the polysaccharide polymerase (20).  
         [0153]    The protein encoded by the  cps 1 I  gene showed some similarity with the Cps14J protein of  S. pneumoniae  (19). The  cps 14 J  gene was shown to encode a β-1, 4-galactosyltransferase activity, responsible for the addition of the fourth (i.e. last) sugar in the synthesis of the  S. pneumoniae  serotype 14 polysaccharide.  
         [0154]    Between Cps1G and Cps1H, a gap of 718 bp was found. This region revealed three small Orfs. The three Orfs were expressed in three different reading frames and were not preceded by potential ribosome binding sites, nor contained potential start sites. However, the three potential gene products encoded by this region showed some similarity with three successive regions of the C-terminal part of the EpsK protein of  Streptococcus thermophilus  (27% identity, 40). The region related to the first 82 amino acids is lacking.  
         [0155]    Analysis of the cloned  cps 9 genes. We also determined the complete nucleotide sequence of the inserts of pCPS9-1 and pCPS9-2 (FIG. 6). Examination of the sequence revealed the presence of three complete and two incomplete Orfs (FIG. 1, part C). As in serotypes 1 and 2, all Orfs are preceded by a ribosome-binding site and are very closely coupled. As suggested by the hybridization data (Table 4), the Cps2D and Cps9D proteins were highly related (Table 2). Based on sequence comparisons, pCPS9-1 lacked the first 27 amino acids of the Cps9D protein.  
         [0156]    The protein encoded by the  cps 9 E  gene showed some similarity with the CapD protein of  Staphylococcus aureus  serotype 1 (24). Based on sequence similarity data, the CapID protein was suggested to be an epimerase or a dehydratase involved in the synthesis of N-acetylfiuctosamine or N-acetylgalactosamine (63).  
         [0157]    Cps9F showed some similarity to the CapM proteins of  S. aureus  serotypes 5 and 8 (61, 64, 65). Based on sequence similarity data, Cap5M and Cap8M are proposed to be glycosyltransferases (63).  
         [0158]    The protein encoded by the  cps 9 G  gene showed some similarity to a protein of  Actinobacillus actinomyceterncomitans  (AB002668 — 4). This protein is part of a gene cluster responsible for the serotype-b specific antigens of  Actinobacillus actinomycetemcomitans.  The function of the protein is unknown.  
         [0159]    The protein encoded by the  cps 9 H  gene showed some similarity to the  rfbB  gene of  Yersinia enterolitica  (68). The RfbB protein was shown to be essential for O-antigen synthesis, but the function of the protein in the synthesis of the 0:3 lipopolysaccharide is unknown.  
         [0160]    Serotype 1 and serotype 9 specific  cps  genes. To determine whether the cloned fragments in pCPS1-1, pCPS1-2, pCPS9-1 and pCPS9-2 contained the type-specific genes for serotype 1 and 9, respectively, cross-hybridization experiments were performed. DNA fragments of the individual  cps 1 and  cps 9 genes were amplified by PCR, labeled with  32 P, and used to probe Southern blots of chromosomal DNA of the reference strains of the 35 different  S. suis  serotypes. The results are shown in Table 5. Based on the data obtained with the  cps 2 E  probe (Table 4), the  cps 1 E  probe was expected to hybridize with chromosomal DNA of  S. suis  serotypes 1, 2, 14, 27 and ½. The  cps 1 H, cps 9 E  and  cps 9 F  probes hybridized with most other serotypes. However, the  cps 1 F  and  cps 1 G  and  cps 1 I  probes hybridized with chromosomal DNA of serotypes 1 and 14 only. The  cps 9 G  and  cps 9 H  probes hybridized with serotype 9 only. These data suggest that the  cps 9 G  and  cps 9 H  probes are specific for serotype 9 and, therefore, could be useful tools for the development of rapid and sensitive diagnostic tests for  S. suis  type 9 infections.  
         [0161]    Type specific PCR. So far, the probes were tested on the 35 different reference strains only. To test the diagnostic value of the typespecific  CpS  probes further, several other  S. suis  serotype 1, 2, ½, 9 and 14 strains were used. Moreover, since a PCR-based method would be even more rapid and sensitive than a hybridization test, we tested whether we could use a PCR for the serotyping of the  S. suis  strains. The oligonucleotide primer sets were chosen within the  cps 2 J, cps 1 I  and  cps 9 H  genes. Amplified fragments of 675 bp, 380 bp and 390 bp were expected, respectively. The results show that 675 bp fragments were amplified on type 2 and ½ strains using  cps 2 J  primers; 380 bp fragments were amplified on type 1 and 14 strains using  cps 1 I  primers and 390 bp fragments were amplified on type 9 strains using  cps 9 H  primers.  
         [0162]    Construction of mutants impaired in capsule production. To evaluate the role of the capsule of  S. suis  type 2 in pathogenesis, we constructed two isogenic mutants in which capsule production was disturbed. To construct mutant 10cpsB, pCPS11 was used. In this plasmid, a part of the  cps 2 B  gene was replaced by the spectinomycin-resistance gene. To construct mutant strain 10cpsEF, the plasmid pCPS28 was used. In pCPS28, the 3′-end of  cps 2 E  gene, as well as the 5′-end, of  cps 2 F  gene, were replaced by the spectinomycin-resistance gene. pCPS 11 and pCPS28 were used to electrotransform strain 10 of  S. suis  type 2 and spectinomycin-resistant colonies were selected. Southern blotting and hybridization experiments were used to select double crossover integration events (results not shown). To test whether the capsular structure of the strains 10cpsB and 10cpsEF was disturbed, we used a slide agglutination test using a suspension of the mutant strains in hyperimmune anti- S. suis  type 2 serum (44). The results showed that even in the absence of serotype specific antisera, the bacteria agglutinated. This indicates that, in the mutant strains, the capsular structure was disturbed. To confirm this, thin sections of wild type and mutant strains were compared by electron microscopy. The results showed that, compared to the wild type (FIG. 3, part A), the amount of capsule produced by the mutant strains was greatly reduced (FIG. 3, part B and part C). Almost no capsular material could be detected on the surface of the mutant strains.  
         [0163]    Capsular mutants are sensitive to phagocytosis and killing by porcine alveolar macrophages (“PAM”). The capsular mutants were tested for their ability to resist phagocytosis by PAM in the presence of porcine SPF serum. The wild type strain 10 seemed to be resistant to phagocytosis under these conditions (FIGS. 9A and 9B). In contrast, the mutant strains were efficiently ingested by macrophages (FIGS. 9A and 9B). After 90 min., more than 99.7% (strain 10cpsB) and 99.8% (strain 10cpsEF) of the mutant cells were ingested by the macrophages. Moreover, as shown in FIGS. 9A and 9B the ingested strains were efficiently killed by the macrophages. 90-98% of all ingested cells were killed within 90 min. No differences could be observed between wild type and mutant strains. These data indicate that the capsule of  S. suis  type 2 efficiently protects the organism from uptake by macrophages in vitro.  
         [0164]    Capsular mutants are less virulent for germfree piglets. The virulence properties of the wild-type and mutant strains were tested after experimental infection of newborn germfree pigs (45, 49). Table 1 shows that specific and nonspecific signs of disease could be observed in all pigs inoculated with the wild type strain. Moreover, all pigs inoculated with the wild type strain died during the course of the experiment or were killed because of serious illness or nervous disorders (Table 3). In contrast, the pigs inoculated with strains 10cpsB and 10cpsEF showed no specific signs of disease and all pigs survived until the end of the experiment (Table 6). The temperature of the pigs inoculated with the wild type strain increased 2 days after inoculation and remained high until day 5 (Table 3). The temperature of the pigs inoculated with the mutant strains sometimes exceeded 40° C., however, we could observe significant differences in the fever index (i.e. percent of observations in an experimental group during which pigs showed fever (&gt;40° C.)) between pigs inoculated with wild type and mutant strains. All pigs showed increased numbers of polymorphonuclear leucocytes (PMLs) (&gt;10×10 9  PMLs per litre) (Table 3). However, in pigs inoculated with the mutant strains, the percentage of samples with increased numbers of PMLs was considerably lower.  S. suis  strains and  B. bronchiseptica  could be isolated from the nasopharynx and feces swab samples of all pigs from 1 day post-infection until the end of the experiment (Table 3). Postmortem, the wild type strain could frequently be isolated from the central nervous system (“CNS”), kidney, heart, liver, spleen, serosae, joints and tonsils. Mutant strains could easily be recovered from the tonsils, but were never recovered from the kidney, liver or spleen. Interestingly, low numbers of the mutant strains were isolated from the CNS, the serosae, the joints, the lungs and the heart. Taken together, these data strongly indicated that mutant  S. suis  strains, impaired in capsule production, are not virulent for young germfree pigs.  
         [0165]    We describe the identification and the molecular characterization of the  cps  locus, involved in the capsular polysaccharide biosynthesis, of  S. suis.  Most of the genes seemed to belong to a single transcriptional unit, suggesting a coordinate control of these genes. We assigned functions to most of the gene products. We thereby identified regions involved in regulation (Cps2A), chain length determination (Cps2B, C), export (Cps2C) and biosynthesis (Cps2E, F, G, H, J, K). The region involved in biosynthesis is located at the center of the gene cluster and is flanked by two regions containing genes with more common functions. The incomplete  orf 2 Z  gene was located at the 5′-end of the cloned fragment. Orf2Z showed some similarity with the YitS protein of  B. subtilis.  However, because the function of the YitS protein is unknown, this did not give us any information about the possible function of Orf2Z. Because the  orf 2 Z  gene is not a part of the cps operon, a role of this gene in polysaccharide biosynthesis is not expected. The Orf2Y protein showed some similarity with the YcxD protein of  B. subtilis  (53). The YcxD protein was suggested to be a regulatory protein. Similarly, Orf2Y may be involved in the regulation of polysaccharide biosynthesis. The Orf2X protein showed similarity with the YAAA proteins of  H. influenzae  and  E. coli.  The function of these proteins is unknown. In  S. suis  type 2, the  orf 2X gene seemed to be the first gene in the  cps 2 operon. This suggests a role of Orf2X in the polysaccharide biosynthesis. In  H. influenzae  and  E. coli,  however, these proteins are not associated with capsular gene clusters. The analysis of isogenic mutants impaired in the expression of Orf2X should give more insight in the presumed role of Orf2X in the polysaccharide biosynthesis of  S. suis  type 2.  
         [0166]    The gene products encoded by the  cps 2 E, cps 2 F, cps 2 G, cps 2 H, cps 2 J  and  cps 2 K  genes showed little similarity with glycosyltransferases of several Gram-positive or Gram-negative bacteria (18, 19, 20, 22, 25). The  cps 2 E  gene product shows some similarity with the  Cps 14 E  protein of  S. pneumoniae  (18, 19). Cps14E is a glucosyl-1-phosphate transferase that links glucose to a lipid carrier (18). In  S. pneumoniae,  this is the first step in the biosynthesis of the oligosaccharide repeating unit. The structure of the  S. suis  serotype 2 capsule contains glucose, galactose, rhamnose, N-acetyl glucosamine and sialic acid in a ratio of 3:1:1:1:1 (7). Based on these data, we conclude that Cps2E of  S. suis  has glucosyltransferase activity and is involved in the linkage of the first sugar to the lipid carrier.  
         [0167]    The C-terminal region of the  cps 2 F  gene product showed some similarity with the RfbU of  Salmonella enteritica.  RfbU was shown to have mannosyltransferase activity (24). Because mannosyl is not a component of the  S. suis  type 2 polysaccharide, a mannosyltransferase activity is not expected in this organism. Nevertheless,  cps 2 F  encodes a glycosyltransferase with another sugar specificity.  
         [0168]    Cps2G showed moderate similarity to a family of gene products suggested to encode galactosyltransferase activities (22, 24, 40). Hence, a similar activity is shown for Cps2G.  
         [0169]    Cps2H showed some similarity with LgtD of  H. influenzae  (U32768). Because LgtD was proposed to have glycosyltransferase activity, a similar activity is fulfilled by Cps2H.  
         [0170]    Cps2J and Cps2K showed similarity to Cps14J of  S. pneumoniae  (20). Cps2J showed similarity with Cps14I of  S. pneumoniae  as well. Cps14I was shown to have N-acetyl glucosaminyltransferase activity, whereas Cps14J has a β-1, 4-galactosyltransferase activity (20). In  S. pneumoniae,  Cps14I is responsible for the addition of the third sugar and Cps14J for the addition of the last sugar in the synthesis of the type 14 repeating unit (20). Because the capsule of  S. suis  type 2 contains galactose as well as N-acetyl glucosamine components, galactosyltransferase as well as N-acetyl glucoaminyltransferase activities could be envisaged for the  cps 2 J  and  cps 2 K  gene products, respectively. As was observed for Cps14I and Cps14J, the N-termini of Cps2J and Cps2K showed a significant degree of sequence similarity. Within the N-terminal domains of Cps14I and Cps14J, two small regions were identified, which were also conserved in several other glycosyltransferases (22). Within these two regions, two Asp residues were proposed to be important for catalytic activity. The two conserved regions, DXS and DXDD, were also found in Cps2J and Cps2K.  
         [0171]    The function of Cps2I remains unclear. Cps2I showed some similarity with a protein of  A. actinomycetemcomitans.  Although this protein part is of the gene cluster responsible for the serotype-B-specific antigens, the function of the protein is unknown.  
         [0172]    We further describe the identification and characterization of the  cps  genes specific for  S. suis  serotypes 1, 2 and 9. After the entire  cps 2 locus of  S. suis  serotype 2 was cloned and characterized, functions for most of the  cps 2 gene products could be assigned by sequence homologies. Based on these data, the glycosyltransferase activities, required for type specificity, could be located in the center of the operon. Cross-hybridization experiments, using the individual  cps 2 genes as probes on chromosomal DNAs of the 35 different serotypes, confirmed this idea. The regions containing the type-specific genes of serotypes 1 and 9 could be cloned and characterized, showing that an identical genetic organization of the  CpS  operons of other  S. suis  serotypes exists. The  cps 1 E, cps 1 F, cps 1 G, cps 1 H ,  and  cps 1 I  genes revealed a striking similarity with  cps 14 E, cps 14 F, cps 14 G, cps 14 H  and  cps 14 J  genes of  S. pneumoniae.  Interestingly,  S. pneumoniae  serotype 14 is the serotype most commonly associated with pneumococcal infections in young children (54), whereas  S. suis  serotype 1 strains are most commonly isolated from piglets younger than 8 weeks (46). In  S. pneumoniae,  the  cps 14 E, cps 14 G, cps 14 I  and cps14 J  encode the glycosyltransferases required for the synthesis of the type 14 tetrameric repeating unit, showing that the  cps 1 E, cps 1 G  and  cps 1 I  genes encoded glycosyltransferases. The precise functions of these genes as well as the substrate specificities of the enzymes can be established. In  S. pneumoniae,  the  cps 14 E  gene was shown to encode a glucosyl-1-phosphate transferase catalyzing the transfer of glucose to a lipid carrier. Moreover,  cpsE -like genes were found in  S. pneumoniae  serotypes 9N, 13, 14, 15B, 15C, 18F, 18A and 19F (60).  CpsE  mutants were constructed in the serotypes 9N, 13, 14 and 15B. All mutant strains lacked glucosyltransferase activity (60). Moreover, in all these  S. pneumoniae  serotypes, the  cpsE  gene seemed to be responsible for the addition of glucose to the lipid carrier. Based on these data, we suggest that in  S. suis  type 1, the  cps 1 E  gene may fulfil a similar function. The structure of the  S. suis  type 1 capsule is unknown, but it is composed of glucose, galactose, N-acetyl glucosamine, N-acetyl galactosamine and sialic acid in a ratio of 1:2.4:1:1:1.4 (5). Therefore, a role of a  cpsE -like glucosyltransferase activity can easily be envisaged.  CpsE -like sequences were also found in serotypes 2, ½ and 14.  
         [0173]    For polysaccharide biosynthesis in  S. pneumoniae  type 14, transfer of the second sugar of the repeating unit to the first lipid-linked sugar is performed by the gene products of  cps 14 F  and  cps 14 G  (20). Similar to  Cps 14 F  and  Cps 14 G,  the  S. suis  type 1 proteins Cps1F and Cps1G may act as one glycosyltransferase performing the same reaction. Cps14F and Cps14G of  S. pneumoniae  showed similarity to the N-terminal half and C-terminal half of the SpsK protein of  Sphingomonas  (20, 67), respectively. This suggests a combined function for both proteins. Moreover,  cps 14 F -and  cps 14 G -like sequences were found in several serotypes of  S. pneumoniae  and these genes always seemed to exist together (60). The same was observed for  S. suis  type 1. The  cps 1 F  and  cps 1 G  probes hybridized with type 1 and type 14 strains.  
         [0174]    According to the similarity found between the cps1 H  gene and the  cps 14 H  gene of  S. pneumoniae  (20),  cps 1 H  is expected to encode a polysaccharide polymerase.  
         [0175]    The protein encoded by the cps1I gene showed some similarity with the  Cps 14 J  protein of  S. pneumoniae  (19). The  cps 14 J  gene was shown to encode a β-1, 4-galactosyltransferase activity, responsible for the addition of the fourth (i.e. last) sugar in the synthesis of the  S. pneumoniae  serotype 14 polysaccharide. In  S. suis  type 2, the proteins encoded by the  cps 2 J  and  cps 2 K  genes showed similarity to the Cps14J protein. However, no significant homologies were found between Cps2J, Cps2K and Cps1I. In the N-terminal regions of Cps14J and Cps14I, two small conserved regions, DXS and DXDD, were identified (19). These regions seemed to be important for catalytic activity (13). At the same positions in the sequence, Cps2I contained the regions DXS and DXED.  
         [0176]    In the region between Cps1G and Cps1H, three small Orfs were identified. Since the Orfs were expressed in three different reading frames, and did not contain potential start sites, expression is not expected. However, the three potential gene products encoded by this region showed some similarity with three successive regions of the C-terminal part of the EpsK protein of  Streptococcus thermophilus  (27% identity, 40). The region related to the first 82 amino acids is lacking. The EpsK protein was suggested to play a role in the export of the exopolysaccharide by rendering the polymerized exopolysaccharide more hydrophobic through a lipid modification. These data could suggest that the sequences in the region between Cps1G and Cps1H originated from epsK-like sequence. Hybridization experiments showed that this  epsK -like region is also present in other serotype 1 strains as well as in serotype 14 strains (results not shown).  
         [0177]    The function of most of the cloned serotype 9 genes can be established. Based on sequence similarity data, the  cps 9 E  and  cps 9 F  genes could be glycosyltransferases (61, 24, 63, 64, 65). Moreover, the  cps 9 G  and  cps 9 H  genes showed similarity to genes located in regions involved in polysaccharide biosynthesis, but the function of these genes is unknown (68).  
         [0178]    Cross-hybridization experiments using the individual  cps 2,  cps 1 and  cps 9 genes as probes showed that the  cps 9 G  and  cps 9 H  probes specifically hybridized with serotype 9 strains.  
         [0179]    Therefore, these are useful as tools for the identification of  S. suis  type 9 strains both for diagnostic purposes as well as in epidemiological and transmission studies. We previously developed a PCR method which can be used to detect  S. suis  strains in nasal and tonsil swabs of pigs (62). The method was used to identify pathogenic (EF-positive) strains of  S. suis  serotype 2. Besides  S. suis  type 2 strains, serotype 9 strains are frequently isolated from organs of diseased pigs. However, until now, a rapid and sensitive diagnostic test was not available for type 9 strains. Therefore, the type 9 specific probes or the type 9 specific PCR is of great diagnostic value. The  cps 1 F, cps 1 G  and  cps 1 I  probes hybridized with serotype 1 as well as with serotype 14 strains. In coagglutination tests, type 1 strains react with the anti-type 1 as well as with the anti-type 14 antisera (56). This suggests the presence of common epitopes between these serotypes. On the other hand, type 1 strains agglutinated only with anti-type 1 serum (56, 57), indicating that it is possible to detect differences between those serotypes.  
         [0180]    The  cps 2 F, cps 2 G, cps 2 H, cps 2 J  and  cps 2 J  probes hybridized with serotypes 2 and ½ only. Serotype 34 showed a weak hybridizing signal with the  cps 2G probe. As shown in agglutination tests, type ½ strains react with sera directed against type 1 as well as with sera directed against type 2 strains (46). Therefore, type ½ shared antigens with both types 1 and 2. Based on the hybridization patterns of serotype ½ strains with the cps1 and  cps 2 specific genes, serotype ½ seemed to be more closely related to type 2 strains than to type 1 strains. In our current studies, we identify type-specific genes, primers or probes which are used for the discrimination of serotypes 1, 14 and 2 and ½ and others of the 35 serotypes yet known. Furthermore, type-specific genes, primers or probes can now easily be developed for yet unknown serotypes, once they become isolated.  
         [0181]    Cloning and characterization of a further part of the cps2 locus.  
         [0182]    Based on the established sequence, 11 genes, designated cps2L to cps2T, orf2U and orf2V, were identified. A gene homologous to genes involved in the polymerization of the repeating oligosaccharide unit (cps20) as well as genes involved in the synthesis of sialic acid (cps2P to cps2T) were identified. Moreover, hybridization experiments showed that the genes involved in the sialic acid synthesis are present in  S. suis  serotypes 1, 2, 14, 27 and ½. The “cps2M” and “cps2N” regions showed similarity to proteins involved in the polysaccharide biosynthesis of other Gram-positive bacteria. However, these regions seemed to be truncated or were nonfunctional as the result of frame-shift or point mutations. At its 3′-end, the cps2 locus contained two insertional elements (“orf2U” and “orf2V”), both of which seemed to be non-functional.  
         [0183]    To clone the remaining part of the cps2 locus, sequences of the 3′-end of pCPS26 (FIG. 1, part C) were used to identify a chromosomal fragment containing cps2 sequences located further downstream. This fragment was cloned in pKUN19, resulting in pCPS29. Using a similar approach, we subsequently isolated the plasmids pCPS30 and pCPS34 containing downstream cps2 sequences (FIG. 1, part C).  
         [0184]    Analysis of the cps2 operon.  
         [0185]    The complete nucleotide sequence of the cloned fragments was determined. Examination of the compiled sequence revealed the presence of: a sequence encoding the C-terminal part of Cps2K, six apparently functional genes (designated cps20-cps2T) and the remnants of 5 different ancestral genes (designated “cps2L”, “cps2M”, “cps2N”, “orf2U” and “orf2V”). The latter genes seemed to be truncated or incomplete as the result of the presence of stop codons or frame-shift mutations (FIG. 1, part A). Neither potential promoter sequences nor potential stem-loop structures could be identified within the sequenced region. A ribosome-binding site precedes each ORF and the majority of the ORFs are very closely linked. Three intergenic gaps were found: one between “cps2M” and “cps2N” (176 nucleotides), one between cps20and cps2P (525 nucleotides), and one between cps2T and “orf2U” (200 nucleotides). These and our above data show that Orf2X and Cps2A-Orf2T are part of a single operon.  
         [0186]    A list of all loci and their properties is shown in Table 4. The “cps2L” region contained three potential ORFs of 103, 79 and 152 amino acids, respectively, which were only separated from each other by stop codons. Only the first ORF is preceded by a potential ribosomal binding site and contained a methionine start codon. This suggests that “cps2L” originates from an ancestral cps2L gene, which coded for a protein of 339 amino acids. The function of this hypothetical Cps2L protein remains unclear so far: no significant homologies were found between Cps2L and proteins present in the data libraries. It is not clear whether the first ORF of the “cps2L” region is expressed into a protein of 103 amino acids. The “cps2M” region showed homology to the N-terminal 134 amino acids of the NeuA proteins of  Streptococcus agalactiae  and  Escherichia coli  (ABO17355, 32). However, although the “cps2 M” region contained a potential ribosome binding site, a methionine start codon was absent. Compared with the  S. agalactiae  sequence, the ATG start codon was replaced by a lysin encoding AAG codon. Moreover, the region homologous to the first 58 amino acids of the  S. agalactiae  NeuA (identity 77%) was separated from the region homologous to amino acids 59-134 of NeuA by a repeated DNA sequence of 100-bp (see, herein). In addition, the region homologous to amino acids 59 to 95 of NeuA (identity 32%) and the region homologous to the amino acids 96 to 134 of NeuA (identity 50%) were present in different reading frames. Therefore, the partial and truncated NeuA homologue is probably nonfunctional in  S. suis . The “cps2N” region showed homology to CpsJ of  S. agalactiae  (accession no. AB017355). However, sequences homologous to the first 88 amino acids of CpsJ were lacking in  S. suis.  Moreover, the homologous region was present in two different reading frames. The protein encoded by the cps2O gene showed homology to proteins of several streptococci involved in the transport of the oligosaccharide repeating unit (accession no. AB017355), suggesting a similar function for Cps2O. The proteins encoded by the cps2P, cps2S and cps2T genes showed homology to the NeuB, NeuD and NeuA proteins of  S. agalactiae  and  E. coli  (accession no. AB017355). Because the “cps2M” region also showed homology to NeuA of  E. coli,  the  S. suis  cps2 locus contains a functional neuA gene (cps2T) as well as a nonfunctional (“cps2M”) gene. The mutual homology between these two regions showed an identity of 77% at the amino acid level over amino acids 1-58 and 49% over the amino acids 59-134. Cps2Q and Cps2R showed homology to the N-terminal and C-terminal parts of the NeuC protein of  S. agalactiae  and  E. coli,  respectively. This suggests that the function of the  S. agalactiae  NeuC protein in  S. suis  is likely fulfilled by two different proteins. In  E. coli,  the neu genes are known to be involved in the synthesis of sialic acid. NeuNAc is synthesized from N-acetylmannosamine and phosphoenolpyruvate by NeuNAc synthetase. Subsequently, NeuNAc is converted to CMP-NeuNAc by the enzyme CMP-NeuNAc synthetase. CMP-NeuNAc is the substrate for the synthesis of polysaccharide. In  E. coli,  K1 NeuB is the NeuNAc synthetase, and NeuA is the CMP-NeuNAc synthetase. NeuC has been implicated in the NeuNAc synthesis, but its precise role is not known. The precise role of NeuD is not known. A role of the Cps2P-Cps2T proteins in the synthesis of sialic acid can easily be envisaged, since the capsule of  S. suis  serotype 2 is rich in sialic acid. In  S. agalactiae,  sialic acid has been shown to be critical to the virulence function of the type III capsule. Moreover, it has been suggested that the presence of sialic acid in the capsule of bacteria which can cause meningitis may be important for these bacteria to breach the blood-brain barrier. So far, however, the requirement of the sialic acid for virulence of  S. suis  remains unclear.  
         [0187]    “Orf2U” and “Orf2V” showed homology to proteins located on two different insertional elements. “Orf2U” is homologous to IS 1194 of  Streptococcus thermophilus,  whereas “Orf2V” showed homology to a putative transposase of  Streptococcus pneumoniae.  This putative transposase was recently found to be associated with the type 2 capsular locus of  S. pneunioniae.  Compared with the original insertional elements in  S. thermophilus  and  S. pneumoniae,  both “Orf2U” and “Orf2V” are likely to be nonfunctional due to frame shift mutations within their coding regions.  
         [0188]    A striking observation was the presence of a sequence of 100 bp (FIG. 10) which was repeated three times within the cps2 operon. The sequence is highly conserved (between 94% and 98%) and was found in the intergenic regions between cps2G and cps2H, within “cps2M” and between cps2O and cps2P. No significant homologies were found between this 100-bp direct repeat sequence and sequences present in the data libraries, suggesting that the sequence is unique for  S. suis.    
         [0189]    Distribution of the cps2 sequences among the 35  S. suis  serotypes.  
         [0190]    To examine the presence of sialic acid encoding genes in other  S. suis  serotypes, we performed cross-hybridization experiments. DNA fragments of the individual cps2 genes were amplified by PCR, radiolabeled with 32P and hybridized to chromosomal DNA of the reference strains of the 35 different  S. suis  serotypes. As a positive control, we used a probe specific for  S. suis  16S rRNA. The 16S rRNA probe hybridized with almost equal intensities to all serotypes tested (Table 4). The “cps2L” sequence hybridized with DNA of serotypes 1, 2, 14 and ½. The “cps2M”, cps20, cps2P, cps2Q, cps2R, cps2S and cps2T genes hybridized with DNA of serotypes 1, 2, 14, 27 and ½. Because the cps2P-cps2T genes are most likely involved in the synthesis of sialic acid, these results suggest that sialic acid is also a part of the capsule in the  S. suis  serotypes 1, 2, 14, 27 and ½. This is in agreement with the finding that the serotypes 1, 2 and ½ possess a capsule that is rich in sialic acid. Although the chemical compositions of the capsules of serotypes 14 and 27 are unknown, recent agglutination studies using sialic acid-binding lectins suggested the presence of sialic acid in  S. suis  serotype 14, but not in serotype 27. In these studies, sialic acid was also detected in serotypes 15 and 16. Since the latter observation is not in agreement with our hybridization studies, it might be that other genes, not homologous to the cps2P-cps2T genes, are responsible for the sialic acid synthesis in serotypes 15 and 16.  
         [0191]    A probe based on “cps2N” sequences hybridized with DNA from serotypes 1, 2, 14 and ½. A probe specific for “orf2U” hybridized with serotypes 1, 2, 7, 14, 24, 27, 32, 34, and ½, whereas a probe specific for “orf2V” hybridized with many different serotypes. In addition, we prepared a probe specific for the 100-bp direct repeat sequence. This probe hybridized with the serotypes 1, 2, 13, 14, 22, 24, 27, 29, 32, 34 and ½ (Table 4). To analyze the number of copies of the direct repeat sequence within the  S. suis  serotype 2 chromosome, a Southern blot hybridization and analysis was performed. Therefore, chromosomal DNA of  S. suis  serotype 2 was digested with NcoI and hybridized with a 32P-labeled direct repeat sequence. Only one hybridizing fragment, containing the three direct repeats present on the cps2 locus, was found (results not shown). This indicates that the 100-bp direct repeat sequence is only associated with the cps2 locus. In  S. pneumoniae,  a 115-bp long repeated sequence was found to be associated with the capsular genes of serotypes 1, 3, 14 and 19F. In  S. pneumoniae,  this 115-bp sequence was also found in the vicinity of other genes involved in pneumococcal virulence (hyaluronidase and neuraminidase genes). A regulatory role of the 11 5-bp sequence in coordinate control of these virulence-related genes was suggested.  
         [0192]    To study the role of the capsule in resistance to phagocytosis and in virulence, we constructed two isogenic mutants in which capsule synthesis was disturbed. In 10cpsB, the  cps 2 B  gene was disturbed by the insertion of an antibiotic-resistance gene, whereas in 10cpsEF, parts of the  cps 2 E  and  cps 2 F  genes were replaced. Both mutant strains seemed to be completely unencapsulated. Because the cps2 genes seemed to be part of an operon, polar effects cannot be excluded. Therefore, these data did not give any information about the role of Cps2B, Cps2E or Cps2F in the polysaccharide biosynthesis. However, the results clearly show that the capsular polysaccharide of  S. suis  type 2 is a surface component with antiphagocytic activity. In vitro wild type encapsulated bacteria are ingested by phagocytes at a very low frequency, whereas the mutant unencapsulated bacteria are efficiently ingested by porcine macrophages. Within 2 hours, over 99.6% of mutant bacteria were ingested and over 92% of the ingested bacteria were killed. Intracellularly, wild type as well as mutant strains seemed to be killed with the same efficiency. This suggests that the loss of capsular material is associated with loss of capacity to resist uptake by macrophages. This loss of resistance to in vitro phagocytosis was associated with a substantial attenuation of the virulence in germfree pigs. All pigs inoculated with the mutant strains survived the experiment and did not show any specific clinical signs of disease. Only some aspecific clinical signs of disease could be observed. Moreover, mutant bacteria could be reisolated from the pigs. This supports the idea that, as in other pathogenic Streptococci, the capsule of  S. suis  acts as an important virulence factor. Transposon mutants prepared by Charland impaired in the capsule production showed a reduced virulence in pigs and mice. To construct these mutants, the type 2 reference strain S735 was used. We previously showed that this strain is only weakly virulent for young pigs. Moreover, the insertion site of the transposon is unsolved so far.  
         [0193]    As a further example herein, a rapid PCT test for  Streptococcus suis type  7 is described.  
         [0194]    Recent epidemiological studies on  Streptococcus suis  infections in pigs indicated that, besides serotypes 1, 2 and 9, serotype 7 is also frequently associated with diseased animals. For the latter serotype, however, no rapid and sensitive diagnostic methods are available. This hampers prevention and control programs. Here we describe the development of a type-specific PCR test for the rapid and sensitive detection of  S. suis  serotype 7. The test is based on DNA sequences of capsular (cps) genes specific for serotype 7. These sequences could be identified by cross-hybridization of several individual cps genes with the chromosomal DNAs of 35 different  S. suis  serotypes.  
         [0195]    [0195] Streptococcus suis  is an important cause of meningitis, septicemia, arthritis and sudden death in young pigs (69, 70). It can, however, also cause meningitis in man (71). Attempts to control the disease are still hampered by the lack of sufficient knowledge about the epidemiology of the disease and the lack of effective vaccines and sensitive diagnostics.  
         [0196]    [0196] S. suis  strains can be identified and classified by their morphological, biochemical and serological characteristics (70, 73, 74). Serological classification is based on the presence of specific antigenic determinants. Isolated and biochemically characterized  S. suis  cells are agglutinated with a panel of specific sera. These typing methods are very laborious and time-consuming and can only be performed on isolated colonies. Moreover, it has been reported that nonspecific cross-reactions may occur among different types of  S. suis  (75, 76).  
         [0197]    So far, 35 different serotypes have been described (7, 78, 79).  S. suis  serotype 2 is the most prevalent type isolated from diseased pigs, followed by serotypes 9 and 1. However, recently, serotype 7 strains were also frequently isolated from diseased pigs (80, 81, 82). This suggests that infections with  S. suis  serotype 7 strains seem to be an increasing problem. Moreover, the virulence of  S. suis  serotype 7 strains was confirmed by experimental infection of young pigs (83).  
         [0198]    Recently, rapid and sensitive PCR assays specific for serotypes 2 (and ½), 1 (and 14) and 9 were developed (84). These assays were based on the cps loci of  S. suis  serotypes 2, 1 and 9 (84, 85). However, until now, no rapid and sensitive diagnostic test was available for  S. suis  serotype 7. Herein we describe the development of a PCR test for the rapid and sensitive detection of  S. suis  serotype 7 strains. The test is based on DNA sequences which form a part of the cps locus of  S. suis  serotype 7. Compared with the serological serotyping methods, the PCR assay was a rapid, reliable and sensitive assay. Therefore, this test, in combination with the PCR tests which we previously developed for serotypes 1, 2 and 9, will undoubtedly contribute to a more rapid and reliable diagnosis of  S. suis  and may facilitate control and eradication programs.  
         [0199]    Materials and Methods  
         [0200]    Bacterial strains, growth conditions and serotyping.  
         [0201]    The bacterial strains and plasmids used in this study are listed in Table 7. The  S. suis  reference strains were obtained from M. Gottschalk, Canada.  S. suis  strains were grown in Todd-Hewitt broth (code CM189, 0xoid), and plated on Columbia agar blood base (code CM331, Oxoid) containing 6% (v/v) horse blood.  E. Coli  strains were grown in Luria broth (86) and plated on Luria broth containing 1.5% (w/v) agar. If required, ampicillin was added to the plates. The  S. suis  strains were serotyped by the slide agglutination test with serotype-specific antibodies (70).  
         [0202]    DNA techniques.  
         [0203]    Routine DNA manipulations and PCR reactions were performed as described by Sambrook et al. (88). Blotting and hybridization were performed as described previously (84, 86).  
         [0204]    DNA sequence analysis.  
         [0205]    DNA sequences were determined on a 373A DNA Sequencing System (Applied Biosystems, Warrington, GB). Samples were prepared by use of an ABI/PRISM dye terminator cycle sequencing ready reaction kit (Applied Biosystems). Custom-made sequencing primers were purchased from Life Technologies. Sequencing data were assembled and analyzed using the McMollyTetra program. The BLAST program was used to search for protein sequences homologous to the deduced amino acid sequences.  
         [0206]    PCR.  
         [0207]    The primers used for the cps7H PCR correspond to the positions 3334-3354 and 3585-3565 in the  S. suis  cps7 locus. The sequences were: 5′-AGCTCTAACACGAAATAAGGC-3′ (SEQ. ID. No. 7) and 5′-GTCAAACACCCTGGATAGCCG3′ (SEQ. ID. No. 8). The reaction mixtures contained 10 mM Tris-HCI, pH 8.3; 1.5 mnM MgC12; 50 mM KC1; 0.2 mM of each of the four deoxynucleotide triphosphates; 1 microM of each of the primers and 1U of AmpliTaq Gold DNA polymerase (Perkin Elmer Applied Biosystems, N.J.). DNA amplification was carried out in a Perkin Elmer 9600 thermal cycler and the program consisted of an incubation for 10 min at 95° C. and 30 cycles of 1 min at 95° C., 2 min at 56° C. and 2 min at 72° C.  
         [0208]    Results and discussion  
         [0209]    Cloning of the seroytpe 7-specific cps genes.  
         [0210]    To isolate the type-specific cps genes of  S. suis  serotype 7, we used the cps9E gene of serotype 9 as a probe to identify chromosomal DNA fragments of type 7 containing homologous DNA sequences (84). A 1.6-kb PstI fragment was identified and cloned in pKUN19. This yielded pCPS7-1 (FIG. 11, part C). In turn, this fragment was used as a probe to identify an overlapping 2.7 kb Scal-Clal fragment. pGEM7 containing the latter fragment was designated pCPS7-2 (FIG. 11, part C).  
         [0211]    Analysis of the cloned cps7 genes.  
         [0212]    The complete nucleotide sequences of the inserts of pCPS7-1, pCPS7-2 were determined. Examination of the cps7 sequence revealed the presence of two complete and two incomplete open reading frames (ORFS) (FIG. 11, part C). All ORFs are preceded by a ribosome-binding Site. In accord with the data obtained for the cps1,  cps 2 and cps9 genes of serotypes 1, 2 and 9, respectively, the type 7 ORFs are very closely linked to each other. The only significant intergenic gap was that found between cps7E and cps7F (443 nucleotides). No obvious promoter sequences or potential stem-loop structures were found in this region. This suggests that, as in serotypes 1, 2 and 9, the cps genes in serotype 7 form part of an operon.  
         [0213]    An overview of the ORFs and their properties is shown in Table 8. As expected on the basis of the hybridization data (84), the Cps9E and Cps7E proteins showed a high similarity (identity 99%, Table 8). Based on sequence comparisons between Cps9E and Cps7E, the PstI fragment of pCPS7-1 lacks the region encoding the first 371 codons of Cps7E. The C-terminal part of the protein encoded by the cps7F gene showed some similarity with the BpIG protein of  Bordetella pertussis  (88), as well as with the C-terminal part of  S. suis  Cps2E (85). Both Bp1G and Cps2E were suggested to have glycosyltransferase activity and are probably involved in the linkage of the first sugar to the lipid carrier (85, 88). The protein encoded by the cps7G gene showed similarity with the BpIF protein of  Bordetella pertussis  (88). BlpF is likely to be involved in the biosynthesis of an amino sugar, suggesting a similar function for Cps7G. The protein encoded by the cps7H gene showed similarity with the WbdN protein of  E. coli  (89) as well as with the N-terminal part of the Cps2K protein of  S. suis  (81). Both WbdN and Cps2K were suggested to have glycosyltransferase activity (85, 89).  
         [0214]    Serotype 7 specific cps genes.  
         [0215]    To determine whether the cloned fragments in pCPS7-1 and pCPS7-2 contained serotype 7-specific DNA sequences, cross-hybridization experiments were performed. DNA fragments of the individual cps7 genes were amplified by PCR, labeled with 32P, and used to probe spot blots of chromosomal DNA of the reference strains of 35 different  S. suis  serotypes. The results are summarized in Table 9. As expected, based on the data obtained with the cps9E probe (84), the cps7E probe hybridized with chromosomal DNA of many different  S. suis  serotypes. The cps7F and cps7G probes showed hybridization with chromosomal DNA of  S. suis  serotypes 4, 5, 7, 17, and 23. However, the cps7H probe hybridized with chromosomal DNA of serotype 7 only, indicating that this gene is specific for serotype 7.  
         [0216]    Type specific PCR.  
         [0217]    We tested whether we could use PCR instead of hybridization for the typing of the  S. suis  serotype 7 strains. For that purpose, we selected an oligonucleotide primer set within the cps7H gene with which an amplified fragment of 251-bp was expected. In addition, we included in our analysis several  S. suis  serotype 7 strains, other than the reference strain. These strains were obtained from different countries and were isolated from different organs (Table 7). The results show that indeed a fragment of about 250-bp was amplified with all type 7 strains used (FIG. 12, part B), whereas no PCR products were obtained with serotype 1, 2 and 9 strains (FIG. 12, part A). This suggests that the PCR test, as described here, is a rapid diagnostic tool for the identification of  S. suis  serotype 7 strains. Until now, such a diagnostic test was not available for serotype 7 Strains. Together with the recently developed PCR assays for serotypes 1, 2, ½, 14 and 9, this assay may be an important diagnostic tool to detect pigs carrying serotype 2, ½, 1, 14, 9 and 7 strains and may facilitate control and eradication programs.  
                             TABLE 1                           Bacterial strains and plasmids                relevant           strain/plasmid   characteristics   source/reference               Strain                 E. coli         CC118   PhoA   (28)       XL2 blue   Stratagene         E. coli         XL2 blue   Stratagene         S. suis         10   virulent serotype 2 strain   (49)       3   serotype 2   (63)       17   serotype 2   (63)       735   reference strain serotype 2   (63)       T15   serotype 2   (63)       6555   reference strain serotype 1   (63)       6388   serotype 1   (63)       6290   serotype 1   (63)       5637   serotype 1   (63)       5673   serotype 1/2   (63)       5679   serotype 1/2   (63)       5928   serotype 1/2   (63)       5934   serotype 1/2   (63)       5209   reference strains serotype 1/2   (63)       5218   reference strain serotype 9   (63)       5973   serotype 9   (63)       6437   serotype 9   (63)       6207   serotype 9   (63)       reference strains   serotypes 1-34   (9, 56, 14)         S. suis         10   virulent serotype 2 strain   (51)       10cpsB   isogenic cpsB mutant of strain 10   this work       10cpsEF   isogenic cpsEF mutant of strain 10   this work       Plasmid       pKUN19   replication functions pUC, Amp R     (23)       pGEM7Zf(+)   replication functions pUC, Amp R     Promega Corp.       pIC19R   replication functions pUC, Amp R     (29)       pIC20R   replication functions pUC, Amp R     (29)       pIC-spc   pIC19R containing spc R  gene   labcollection           of pDL282       pDL282   replication functions of pBR322   (43)           and pVT736-1, Amp R , Spc R         pPHOS2   pIC-spc containing the truncated   this work           phoA gene of pPHO7 as a           PstI-BamHI fragment       pPHO7   contains truncated phoA gene   (15)       pPHOS7   pPHOS2 containing chromosomal   this work             S. suis  DNA       pCPS6   pKUN19 containing 6 kb HindIII   this work (FIG. 1)           fragment of cps operon       pCPS7   pKUN19 containing 3,5 kb EcoRI-   this work (FIG. 1)           HindIII fragment of cps operon       pcPS11   pCPS7 in which 0.4 kb PstI-   this work (FIG. 1)           BamHI fragment of cpsB gene is           replaced by Spc R  gene           of pIC-spc       pCPS17   pKUN19 containing 3.1 kb KpnI   this work (FIG. 1)           fragment of cps operon       pCPS18   pKUN19 containing 1.8 kb SnaBI   this work (FIG. 1)           fragment of cps operon       pCPS20   pKUN19 containing 3.3 kb XbaI-   this work (FIG. 1)           HindIII fragment of cps operon       pCPS23   pGEM7Zf(+) containing 1.5 kb   this work (FIG. 1)           M1uI fragment of cps operon       pCPS25   pIC20R containing 2.5 kb   this work (FIG. 1)           KpnI-Sa1I fragment of pCPS17       pCPS26   pKUN19 containing 3.0 kb HindIII   this work (FIG. 1)           fragment of cps operon       pCPS27   pCPS25 containing 2.3 kb XbaI   this work (FIG. 1)           (blunt)-C1aI fragment of pCPS20       pCPS28   pCPS27 containing the 1.2 kb   this work (FIG. 1)           PstI-XhoI Spc R  gene of           pIC-spc       pCPS29   pKUN19 containing 2.2 kb SacI-   this work (FIG. 1)           PstI fragment of cps operon       pCPS1-1   pKUN19 containing 5 kb EcoRV   this work (FIG. 1)           fragment of cps operon of type 1       pCPS1-2   pKUN19 containing 2.2 kb HindIII   this work (FIG. 1)           fragment of cps operon of type 1       pCPS9-1   pKUN19 containing 1 kb HindIII-   this work (FIG. 1)           XbaI fragment of cps operon of           serotype 9       pCPS9-2   pKUN19 containing 4.0 kb   this work (FIG. 1)           XbaI-XbaI fragment of cps operon           of serotype 9                                          
 
         [0218]    [0218]                                         TABLE 2                           Properties of Orfs in the cps locus of  S. suis  serotype 2 and similarities to gene product other bacteria                nucleotide   number                       position in   of amino       proposed function   similar gene product       ORF   sequence   acids   GC %   of gene product 1     (% identity)               Orf2Z   1-719   240   44   Unknown     B. subtilis  YitS (26%)       Orf2Y   2079-822   419   38   Transcription     B. subtilis  YcxD (39%)                       regulation       Orf2X   2202-2934   244   39   Unknown     H. influenzae  YAAA (24%)       Cps2A   3041-4484   481   39   Regulation     S. pneumoniae  Cps19fA (58%)       Cps2B   4504-5191   229   40   Chain length     S. pneumoniae  type 3 Orfl (58%)                       determination       Cps2C   5203-5878   225   40   Chain length     S. pneumoniae  Cps23fD (63%)                       determination/                       Export       Cps2D   5919-6648   243   38   Unknown     S. pneumoniae  CpsB (62%)       Cps2E   6675-8052   459   33   Glycosyltransferase     S. pneumoniae  Cps14E (56%)       Cps2F   8089-9256   389   32   Glycosyltransferase     S. pneumoniae  Cps23fT       Cps2G   9262-10417   385   36   Glycosyltransferase     S. thermophilus  EpsF (25%)       Cps2H   10808-12176   457   31   Glycosyltransferase     S. mutans  RGPEC, N  (29%)       Cps2I   12213-13443   410   29   CP polymerase     S. pneumoniae  Cps23fI (48%)       Cps2J   13583-14579   332   29   Glycosyltransferase     S. pneumoniae  Cps14J (31%)       Cps2K   14574-15576   334   37   Glycosyltransferase     S. pneumoniae  Cps14J (40%)       “Cps2L”   15618-16635   103   37   Unknown   —       “Cps2M”   16811-17322   —   38   —     S. agalactiae  CpsF N  (77%)                             E. coli  NeuA , N  (47%)       “Cps2N”   17559-18342   —   39   —     S. agalactiae  CpsJ (43%)       Cps2O   18401-19802   476   40   Repeat unit     S. agalactiae  CpsK (41%)                       transporter       Cps2P   20327-21341   338   39   Sialic acid synthesis     S. agalactiae  NeuB (80%)                             E. coli  NeuB (59%)       Cps2Q   21355-21865   170   42   Sialic acid synthesis     S. agalactiae  NeuC N  (61%)                             E. coli  NeuC N  (54%)       Cps2R   21933-22483   184   40   Sialic acid synthesis     S. agalactiae  NeuC c  (55%)                             E. coli  NeuC c  (40%)       Cps2S   22501-23125   208   42   Sialic acid synthesis     E. coli  NeuD (32%)       Cps2T   23136-24366   395   40   CMP-NeuNAc     S. agalactiae  CpsF (49%)                       synthetase     E. coli  NeuA (34%)       “Orf2U”   24566-25488   168   42   Transposase     S. thermophilus  IS1194 (51%)       “Orf2V”   25691-26281   116   37   Transposase     S. pneumoniae  orf1 (85%)                                                    
         [0219]    [0219]                                                                               TABLE 3                           Properties of Orfs in the cps genes of  S. suis  serotypes 1 and 9       and similarities to gene products of other bacteria                nucleotide       number   (kDa)                    position in       of amino   predicted   predicted   proposed function   similar gene product   reference/       ORF   sequence   G + C%   acids   mol. mass   pI   of gene product 1     (% identity)   accession nr.               Cps1E 2     1-1363   34%   454   52.2   8.0   Glucosyltransferase     Streptococcus suis  Cps2E                                       (86%)   (26)                                     Streptococcus pneumoniae  Cps14E                                   (48%)   (12)       Cps1F   1374-1821   33%   149   17.3   8.2   Unknown     Streptococcus pneumoniae  Cps14F                                   (83%)   (14)       Cps1G   1823-2315   25%   164   19.5   7.5   Glycosyltransferase     Streptococcus pneumoniae  Cps14G                                   (50%)   (14)       Cps1H   3035-4202   24%   389   45.5   8.4   CP polymerase     Streptococcus pneumoniae  Cps14H                                   (30%)   (14)       Cps1I   4197-                   Glycosyltransferase     Streptococcus pneumoniae  Cps14J                                   (38%)   (13)                                     Lactoccocus lactis  EpsG                                   (31%)   (29)                                     Streptococcus thermophilus  EpsI                                   (33%)   (28)       Cps1J                       Glycosyltransferase   Streptococcus pneumoniae Cps14J                                   ( )   (13)       Cps1K 3         37%   278   32.5   7.8   Glycosyltransferase     Streptococcus pneumoniae  Cps14J                                   (44%)   (13)       Cps9D 2     1-646   37%   215   24.9   8.1   Unknown     Streptococcus suis  Cps2D                                   (89%)   (26)       Cps9E   680-                   Glycosyltransferase     Staphylococcus aureus  Cap1D                                   (27%)   (18)       Cps9F       36%   200   22.3   8.2   Glycosyltransferase     Staphylococcus aureus  Cap5M                                   (52%)   (17)       Cps9G       35%   269   31.5   8.0   Unknown     Actinobacillus actinomycetemcomitans                                     (43%)   (A8002668_4)                                     Haemophilus influenzae  Lsg                                   (43%)   (005081)       Cps9H 3         30%   143   16.5   7.2   Unknown     Yersinia enterolitica  RfbB                                   (28%)   (33)                                            
         [0220]    [0220]                                                                                                                                                                                           TABLE 4                       Hybridization of serotype 2 cps genes and neighboring       sequences with chromosomal DNA of other serotypes                                    serotypes            DNA probes   1   2   3   4   5   6   7   8   9   10   11   12   13   14   15   16   17   18               orf2Z   +   +   +   +   +   +   +   +   +   +   +   +   ±   +   +   +   +   +       orf2Y   +   +   +   +   +   +   +   +   +   +   +   +   ±   +   +   +   +   +       orf2X   +   +   +   +   +   +   +   +   +   +   +   +   ±   +   +   +   +   +       cps2A   +   +   +   +   +   +   +   +   +   +   +   +   +   +   +   +   +   +       cps2B   +   +   +   +   +   +   +   +   +   +   −   −   ±   +   −   −   ±   ±       cps2C   +   +   +   +   +   +   +   +   +   +   +   −   ±   +   −   ±   −   −       cps2D   +   +   +   +   +   +   +   +   +   +   +   ±   ±   +   −   ±   +   +       cps2E   +   +   −   −   −   −   −   −   −   −   −   −   −   +   −   −   −   −       cps2F   −   +   −   −   −   −   −   −   −   −   −   −   −   −   −   −   −   −       cps2G   −   +   −   −   −   −   −   −   −   −   −   −   −   −   −   −   −   −       cps2H   −   +   −   −   −   −   −   −   −   −   −   −   −   −   −   −   −   −       cps2I   −   +   −   −   −   −   −   −   −   −   −   −   −   −   −   −   −   −       cps2J   −   +   −   −   −   −   −   −   −   −   −   −   −   −   −   −   −   −       cps2K   +   +   −   −   −   −   −   −   −   −   −   −   −   +   −   −   −   −       “cps2L”   +   +   −   −   −   −   −   −   −   −   −   −   −   +   −   −   −   −       “cps2M”   +   +   −   −   −   −   −   −   −   −   −   −   −   +   −   −   −   −       “cps2N”   +   +   −   −   −   −   −   −   −   −   −   −   −   +   −   −   −   −       cps2O   +   +   −   −   −   −   −   −   −   −   −   −   −   +   −   −   −   −       cps2P   +   +   −   −   −   −   −   −   −   −   −   −   −   +   −   −   −   −       cps2Q   +   +   −   −   −   −   −   −   −   −   −   −   −   +   −   −   −   −       cps2R   +   +   −   −   −   −   −   −   −   −   −   −   −   +   −   −   −   −       cps2S   +   +   −   −   −   −   −   −   −   −   −   −   −   +   −   −   −   −       cps2T   +   +   −   −   −   −   −   −   −   −   −   −   −   +   −   −   −   −       “orf2U”   +   +   −   −   −   −   +   −   −   −   −   −   −   +   −   −   −   −       “orf2V”   +   +   ±   ±   ±   −   ±   −   −   −   −   −   −   +   +   −   +   +       100-bp repeat   +   +   −   −   −   −   −   −   −   −   −   −   +   +   −   −   −   −       16SrRNA   +   +   +   +   +   +   +   +   +   +   +   +   +   +   +   +   +   +                        serotypes            DNA probes   19   20   21   22   23   24   25   26   27   28   29   30   31   32   33   34   ½               orf2Z   +   −   +   −   +   +   +   −   +   +   +   +   +   −   −   −   +       orf2Y   +   ±   +   ±   +   +   +   +   +   +   +   +   +   −   −   −   +       orf2X   +   −   +   −   +   +   +   −   +   +   +   +   +   −   −   −   +       cps2A   +   −   +   −   +   +   +   −   +   +   +   +   +   −   −   −   +       cps2B   ±   −   ±   −   +   +   +   −   −   −   +   ±   +   −   ±   −   +       cps2C   −   −   −   −   +   +   +   −   +   ±   −   −   +   −   ±   −   +       cps2D   +   −   ±   −   +   +   +   −   +   +   +   ±   +   −   −   −   +       cps2E   −   −   −   −   −   −   −   −   +   −   −   −   −   −   −   −   +       cps2F   −   −   −   −   −   −   −   −   −   −   −   −   −   −   −   −   +       cps2G   −   −   −   −   −   −   −   −   −   −   −   −   −   −   −   ±   +       cps2H   −   −   −   −   −   −   −   −   −   −   −   −   −   −   −   −   +       cps2I   −   −   −   −   −   −   −   −   −   −   −   −   −   −   −   −   +       cps2J   −   −   −   −   −   −   −   −   −   −   −   −   −   −   −   −   +       cps2K   −   −   −   −   −   −   −   −   −   −   −   −   −   −   −   −   +       “cps2L”   −   −   −   −   −   −   −   −   −   −   −   −   −   −   −   −   +       “cps2M”   −   −   −   −   −   −   −   −   +   −   −   −   −   −   −   −   +       “cps2N”   −   −   −   −   −   −   −   −   −   −   −   −   −   −   −   −   +       cps2O   −   −   −   −   −   −   −   −   +   −   −   −   −   −   −   −   +       cps2P   −   −   −   −   −   −   −   −   +   −   −   −   −   −   −   −   +       cps2Q   −   −   −   −   −   −   −   −   +   −   −   −   −   −   −   −   +       cps2R   −   −   −   −   −   −   −   −   +   −   −   −   −   −   −   −   +       cps2S   −   −   −   −   −   −   −   −   +   −   −   −   −   −   −   −   +       cps2T   −   −   −   −   −   −   −   −   +   −   −   −   −   −   −   −   +       “orf2U”   −   −   −   −   −   +   −   −   +   −   −   −   −   +   −   +   +       “orf2V”   ±   −   −   ±   +   −   −   +   −   −   −   −   +   +   −   ±   +       100-bp repeat   −   −   −   +   −   +   −   −   +   −   −   −   −   +   −   +   +       16SrRNA   +   +   +   +   +   +   +   +   +   +   +   +   +   +   +   +   +                    
         [0221]    [0221]                                                                                                                     TABLE 5                           Hybridization of serotypes 1 and 9 cps genes       with chromosomal DNA of other  S. suis  serotypes                DNA probes            Serotype   cps1E   cps1F   cps1G   cps1H   cps1I   cps9E   cps9F   cps9G   cps9H   16rRNA                    1   +   +   +   +   +   −   −   −   −   +       2   +   −   −   −   −   −   −   −   −   +       3   −   −   −   +   −   +   −   −   −   +       4   −   −   −   +   −   +   −   −   −   +       5   −   −   −   +   −   +   −   −   −   +       6   −   −   −   −   −   −   −   −   −   +       7   −   −   −   +   −   +   −   −   −   +       8   −   −   −   −   −   −   −   −   −   +       9   −   −   −   +   −   +   +   +   +   +       10   −   −   −   +   −   +   +   −   −   +       11   −   −   −   +   −   +   ±   −   −   +       12   −   −   −   ±   −   +   ±   −   −   +       13   −   −   −   +   −   +   −   −   −   +       14   +   +   +   +   +   −   −   −   −   +       15   −   −   −   −   −   −   −   −   −   +       16   −   −   −   −   −   −   −   −   −   +       17   −   −   −   +   −   +   −   −   −   +       18   −   −   −   +   −   +   −   −   −   +       19   −   −   −   +   −   +   −   −   −   +       20   −   −   −   −   −   −   −   −   −   +       21   −   −   −   +   −   +   ±   −   −   +       22   −   −   −   −   −   −   −   −   −   +       23   −   −   −   +   −   +   −   −   −   +       24   −   −   −   +   −   +   +   −   −   +       25   −   −   −   −   −   −   −   −   −   +       26   −   −   −   −   −   −   ±   −   −   +       27   +   −   −   −   −   −   −   −   −   +       28   −   −   −   +   −   +   ±   −   −   +       29   −   −   −   +   −   +   −   −   −   +       30   −   −   −   +   −   +   ±   −   −   +       31   −   −   −   +   −   +   −   −   −   +       32   −   −   −   −   −   −   −   −   −   +       33   −   −   −   −   −   −   ±   −   −   +       34   −   −   −   −   −   −   −   −   −   +       ½   +   −   −   −   −   −   −   −   −   +                    
         [0222]    [0222]                                                                                                                                                                               TABLE 6                           Virulence of wild type and capsular mutant  S. suis  strains in germfree pigs                        clinical index of           isolation of           pigs/       the group       leuco−     S. suis  in pigs              S. suis     group   mortality 2     morbidity 3     spec   non-spec.   fever   cyte   [n] per group in            strains 1     [n]   [%]   [%]   symptoms 5     symptoms 6     index 7     index 8     CNS   serosae   joints                    10   4   100   100   11   88   43   44   2   3   4       10cpsB   4   0   0   0   10   1   3   1   3   2       10cpsEF   4   0   0   0   0   1   0   1   3   2                                                                                    
         [0223]    [0223]                         TABLE 7                           Bacterial strains and plasmids            strain/plasmid   relevant characteristics               Strain             E. coli         XL2 blue         S. suis         reference strains   serotypes 1-34       5667   serotype 7, tonsil (1993)       7037   serotype 7, organs (1994)       7044   serotype 7, brains (1994)       7068   serotype 7 (1994)       7646   serotype 7 (1994)       7744   serotype 7, lungs (1996)       7759   serotype 7, joints (1996)       8169   serotype 7 (1997)       15913   serotype 7, meninges (1998)       Plasmid       pKUN19replication       functions pUC, Amp R         pGEM7Zf(+)   replication functions pUC, Amp R         pCPS9-1   pKUN19 containing 1 kb HindIII-XbaI           fragment of cps operon of serotype 9       pCPS9-2   pKUN19 containing 4.0 kb XbaI-XbaI           fragment of cps operon of serotype 9       pCPS7-1   pKUN19 containing 1.6-kb PstI fragment           of cps operon of type 7       pCPS7-2   pGEM7 containing 2.7-kb ScaI-C1aI fragment           of cps operon of type 7                                    
         [0224]    [0224]                                 TABLE 8                           Properties of Orfs in the cps genes of  S. suis  serotype 7 and       similarities to gene products of other bacteria                nucleotide                   position in   proposed function   similar gene product       Orf   sequence   of gene product   (% identity)               Cps7E   1-719   Glycosyltransferase     Streptococcus suis                     Cps9E (99%)       Cps7F   1164-1863   Glycosyltransferase     Bordetella pertussis                     Bp1G 1  (43%)                     Streptococcus suis                     Cps2E 1  (33%)       Cps7G   1872-3086   Biosynthesis amino sugar     Bordetella pertussis                     Bp1F (48%)       Cps7H   3104-3737   Glycosyltransferase     Escherichia coli                     WbdN (35%)                     Streptococcus suis                     Cps2K 2  (31%)                                    
         [0225]    [0225]                                                                                                                                                                                           TABLE 9                       Hybridization of serotype 7 cps probes with chromosomal DNA of  S. suis  serotypes                                    serotypes            DNA probes   1   2   3   4   5   6   7   8   9   10   11   12   13   14   15   16   17   18               cps7E   −   −   +   +   +   −   +   −   +   +   +   +   +   −   −   −   +   +       cps7F   −   −   −   +   +   −   +   −   −   −   −   −   −   −   −   −   +   −       cps7G   −   −   −   +   +   −   +   −   −   −   −   −   −   −   −   −   +   −       cps7H   −   −   −   −   −   −   +   −   −   −   −   −   −   −   −   −   −   −       16SrRNA   +   +   +   +   +   +   +   +   +   +   +   +   +   +   +   +   +   +                serotypes            DNA probes   19   20   21   22   23   24   25   26   27   28   29   30   31   32   33   34   ½               cps7E   +   −   +   −   +   +   −   −   −   −   +   +   +   −   −   −   −       cps7F   −   −   −   −   +   −   −   −   −   −   −   −   −   −   −   −   −       cps7G   −   −   −   −   +   −   −   −   −   −   −   −   −   −   −   −   −       cps7H   −   −   −   −   −   −   −   −   −   −   −   −   −   −   −   −   −       16SrRNA   +   +   +   +   +   +   +   +   +   +   +   +   +   +   +   +   +                    
       REFERENCES  
       [0226]    1. Arends, J. P., and H. C. Zanen. 1988. Meningitis caused by  Streptococcus suis  in humans. Rev. Infect. Dis. 10:131-37.  
         [0227]    2. Arrecubieta, C., E. Garcia, and R. Lopez. 1995. Sequence and transcriptional analysis of a DNA region involved in the production of capsular polysaccharide in  Streptococcus pneumoniae  type 3. Gene 167:1-7  
         [0228]    3. Arrecubieta, C., R. Lopez, and E. Garcia. 1994. Molecular characterization of  cap 3A, a gene from the operon required for the synthesis of the capsule of  Streptococcus pneumoniae  type 3: sequencing of mutations responsible for the unencapsulated phenotype and localization of the capsular cluster on the pneumococcal chromosome. J. Bacteriol. 176:6375-6383.  
         [0229]    4. Clifton-Hadley, F.A. 1983.  Streptococcus suis  type 2 infections. Br. Vet. J. 139:1-5.  
         [0230]    5. Charland, N., J. Harel, N., Kobisch, S. Lacasse, and M. Gottschalk. 1998. Streptococcus suis serotype 2 mutants deficient in capsular expression. Microbiol. 144:325-332.  
         [0231]    6. Cross, A. S. 1990. The biological significance of bacterial encapsulation. Curr. Top. Microbiol. Immunol. 150:87-95.  
         [0232]    7. Elliott, S. D. and J. Y. Tai. 1978. The type specific polysaccharide of  Streptococcus suis.  J. Exp. Med. 148:1699-1704.  
         [0233]    8. Feder, I., M. M. Chengappa, B. Fenwick, M. Rider and J. Staats. 1994. Partial characterization of  Streptococcus suis  type 2 hemolysin. J. Clin. Microbiol. 32:1256-1260.  
         [0234]    9. Gottschalk, M., R. Higgins, M. Jacques, M. Beaudoin, and J. Henrichsen. 1991. Characterization of six new capsular types (23 through 28) of  Streptococcus suis.  J. Clin. Microbiol. 29:2590-2594.  
         [0235]    10. Gottschalk, M., S. Lacouture, and J. D. Dubreuil. 1995. Characterization of  Streptococcus suis  type 2 haemolysin. Microbiology 141:189-195.  
         [0236]    11. Gottschalk, M., A. Lebrun, M. Jacques, and R. Higgins. 1990. Haemagglutination properties of  Streptococcus suis.  J. Clin. Microbiol. 28:2156-2158.  
         [0237]    12. Guidolin, A., J. M. Morona, R. Morona, D. Hansman, and J. C. Paton. 1994. Nucleotide sequence analysis of genes essential for capsular polysaccharide biosynthesis in  Streptococcus pneumoniae  type 19F. 1994. Infect. Immun. 62:5384-5396.  
         [0238]    13. Guitierrez, C., and J. C. Devedjian. 1989. Plasmid facilitating in vitro construction of PhoA fusions in  Escherichia coli.  Nucl. Acid. Res. 17:3999.  
         [0239]    14. Higgins, R., M. Gottschalk, M. Boudreau, A. Lebrun, and J. Henrichsen. 1995. Description of six new capsular types (28 through 34) of  Streptococcus suis.  J. Vet. Diagn. Invest. 7:405-406  
         [0240]    15. Jacobs, A. A., P. L. W. Loeffen, A. J. G. van den Berg, and P. K. Storm. 1994. Identification, purification and characterization of a thiol-activated hemolysin (suilysin) of  Streptococcus suis.  Infect. Immun. 62:1742-1748.  
         [0241]    16. Jacques, M., M. Gottschalk, B. Foiry B. and R. Higgins. 1990. Ultrastructural study of surface components of  Streptococcus  suis.  J. Bacteriol.  172:2833-2838.  
         [0242]    17. Klein P., M. Kanehisa and C. DeLisi. 1985. The detection and classification of membrane spanning proteins. Biochim. Biophys. Acta. 851:468-476.  
         [0243]    18. Kolkman, M. A. B., D. A. Morrison, B. A. M. van der Zeijst, and P. J. M. Nuijten. 1996. The capsule polysaccharide synthesis locus of  Streptococcuspneumoniae serotype  14: identification of the glycosyl transferase gene  cps 14 E.  J. Bacteriol. 178:3736-3541.  
         [0244]    19. Kolkman, M. A. B., W. Wakarchuk, P. J. M. Nuijten, and B. A. M. van der Zeijst. 1997. Capsular polysaccharide synthesis in  Streptococcuspneumoniae serotype  14: molecular analysis of the complete  cps  locus and identification of genes encoding glycosyltransferases required for the biosynthesis of the tetrasaccharide subunit. Mol. Microbiol. 26:197-208.  
         [0245]    20. Kolkinan, M. A. B., B. A. M. van der Zeijst and P. J. M. Nuijten. 1997. Functional analysis of glycosyltransferases encoded by the capsular polysaccharide biosynthesis locus of  Streptococcus pneumoniae  serotype 14. J. Biol. Chem. 272:1950219508.  
         [0246]    21. Konings, R. N. H., E. J. M. Verhoeven, and B. P H. Peeters. 1987. pKUN vectors for the separate production of both DNA strands of recombinant plasmids. Methods Enzymol. 153:12-34.  
         [0247]    22. Korolik, V., B. N. Fry, M. R. Alderton, B. A. M. van der zeijst, and P. J. Coloe. 1997. Expression of  Campylobacter hyoilei  lipo-oligosaccharide (LOS) antigens in  Escherichia coli.  Microbiol. 143:3481-3489.  
         [0248]    23. Leij, P. C. J., R. van Furth, and T. L. van Zwet. 1986. In vitro determination of phagocytosis and intracellular killing of polymorphonuclear and mononuclear phagocytes. In Handbook of Experimental Immunology, vol. 2. Cellular immunology, pp. 46.1-46.21. Edited by D. M. Weir, L. A. Herzenberg, C. Blackwell and L. A. Herzenberg. Blackwell scientific Publications, Oxford.  
         [0249]    24. Lin, W. S., T. Cunneen, and C. Y. Lee. 1994. Sequence analysis and molecular characterization of genes required for the biosynthesis of type 1 capsular polysaccharide in  Staphylococcus aureus.  J. Bacteriol. 176:7005-7016.  
         [0250]    25. Liu, D., A. M. Haase, L. Lindqvist, A.A. Lindberg, and P. R. Reeves. 1993. Glycosyl transferases of O-antigen biosynthesis in  Salmonella enteritica : Identification and characterization of transferase genes of group B, C2, and El. J. Bacteriol. 175:3408-3413.  
         [0251]    26. Manoil, C., and J. Beckwith. 1985. A transposon probe for protein export signals. Proc. Natl. Acad. Sci. USA 82:8129-8133.  
         [0252]    27. Marsh, J. L., M. Erfle, and E. J. Wykes. 1984. The pIC plasmnid and phage vectors with versatile cloning sites for recombinant selection by insertional inactivation. Gene 32:481-485.  
         [0253]    28. Miller, J. 1972. Experiments in Molecular Genetics. Cold Spring Harbor, N.Y.: Cold Spring Harbor Laboratory.  
         [0254]    29. Morona, J. K., R. Morona, and J. C. Paton. 1997. Characterization of the locus encoding the  Streptococcus pneumoniae  type 19F capsular polysaccharide biosynthesis pathway. Mol. Microbiol. 23: 761-763.  
         [0255]    30. Mufioz, R., M. Mollerach, R. L6pez and E. Garcia. 1997. Molecular organization of the genes required for the synthesis of type 1 capsular polysaccharide of  Streptococcus pneumoniae;  formation of binary encapsulated pneumococci and identification of cryptic dTDP-rhamnose biosynthesis genes. Mol. Microbiol. 25:79-92.  
         [0256]    31. Pearce B. J., Y. B. Yin, and H. R. Masure. 1993. Genetic identification of exported proteins in  Streptococcus pneumoniae.  Mol. Microbiol. 9:1037-1050.  
         [0257]    32. Roberts, I. S. 1996. The biochemistry and genetics of capsular polysaccharide production in bacteria. Ann. Rev. Microbiol. 50:285-315.  
         [0258]    33. Rossbach, S., D. A. Kulpa, U. Rossbach, and F. J. de Bruin. 1994. Molecular and genetic characterization of the rhizopine catabolism (mocABRC) genes of  Rhizobium meliloti  L5-30. Mol. Gen. Genet. 245: 11-24.  
         [0259]    34. Rubens, C. E., L. M. Heggen, R. F. Haft, and R. M. Wessels. 1993. Identification of  cpsD,  a gene essential for type III capsule expression in group B streptococci. Mol. Microbiol. 8:843-855.  
         [0260]    35. Rubens, C. E., L. M. R. Wessels, L. M. Heggen, and D. L. Kasper. 1987. Transposon mutagenesis of type III group  B Streptococcus  correlation of capsule expression with virulence. Proc. Natl. Acad. Sci. USA 84:7208-7212.  
         [0261]    36. Sambrook, J., E. F. Fritsch, and T. Maniatis. 1989. Molecular cloning. A laboratory manual. Second edition. Cold Spring Harbor Laboratory Press. Cold Spring Harbor, N.Y.  
         [0262]    37. Smith, H. E., U. Vecht, H. J. Wisselink, N. Stockhofe-Zurwieden, Y. Biermann, and M. A. Smits. 1996. Mutants of  Streptococcus suis  types 1 and 2 impaired in expression of muramidase-released protein and extracellular protein induce disease in newborn germfree pigs. Infect Immun. 64:4409-4412.  
         [0263]    38. Smith, H. E., H. J. Wisselink, U. Vecht, A. L. J. Gielkens and M. A. Smits. 1995. High-efficiency transformation and gene inactivation in  Streptococcus suiss  type 2. Microbiol. 141:181-188.  
         [0264]    39. Sreenivasan, P. K., D. L. LeBlanc, L. N. Lee, and P. Fives-Taylor. 1991. Transformation of  Actinobacillus actinomycetemcomitans  by electroporation, utilizing constructed shuttle plasmids. Infect. Immun. 59:4621-4627.  
         [0265]    40. Stringele F., J.R. Neeser, and B. Mollet. 1996. Identification and characterization of the eps (exopolysaccharide) gene cluster from  Streptococcus thermophilus  Sfi6. J. Bacteriol. 178:1680-1690.  
         [0266]    41. Stockhofe-Zurwieden, N., U. Vecht, H. J. Wisselink, H. van Lieshout, and H. E. Smith. 1996. Comparative studies on the pathogenicity of different  Streptococcus suis  serotype 1 strains. In Proceedings of the 14th IPVS Congress. pp. 299.  
         [0267]    42. van Kranenburg, R., J. D. Marugg, I. I. van Swam, N. J. Willem and W. M. de Vos. 1997. Molecular characterization of the plasinid-encoded  eps  gene cluster essential for exopolysaccharide biosynthesis in  Lactococcus lactis  Mol. Microbiol. 24:387-397.  
         [0268]    43. van Leengoed, L. A., E. M. Kamp, and J. M. A. Pol. 1989. Toxicity of  Haemophilus pleuropneumoniae  to porcine lung macrophages. Vet. Microbiol. 19:337-349.  
         [0269]    44. van Leengoed, L. A. M. G., U. Vecht, and E. R. M. Verheyen. 1987.  Streptococcus suis  type 2 infections in pigs in The Netherlands (part two). Vet Quart. 9, 111-117.  
         [0270]    45. Vecht, U., J. P. Arends, E. J. van der Molen, and L. A. M. G. van Leengoed. 1989. Differences in virulence between two strains of  Streptococcus suis  type 2 after experimentally induced infection of newborn germfree pigs. Am. J. Vet. Res. 50:1037-1043.  
         [0271]    46. Vecht, U., L. A. M. G. van Leengoed, and E. R. M. Verheyen. 1985.  Streptococcus suis  infections in pigs in The Netherlands (part one). Vet. Quart. 7:315-321.  
         [0272]    47. Vecht, U., H. J. Wisselink, M. L. Jellema, and H. E. Smith. 1991. Identification of two proteins associated with virulence of  Streptococcus suis  type 2. Infect. Immun. 59:3156-3162.  
         [0273]    48. Vecht, U., H. J. Wisselink, N. Stockhofe-Zurwieden, and H. E. Smith. 1996. Characterization of virulence of the  Streptococcus suis  serotype 2 reference strain Henrichsen  S.  735 in newborn gnotobiotic pigs. Vet. Microbiol. 51:125-136.  
         [0274]    49. Vecht, U., H. J. Wisselink, J. E. van Dijk, and H. E. Smith. 1992. Virulence of  Streptococcus suis  type 2 strains in newborn germfree pigs depends on phenotype. Infect. Immun. 60:550-556.  
         [0275]    50. Wagenaar, F., G. L. Kok, J. M. Broekhuijsen-Davies, and J. M. A. Pol. 1993. Rapid cold fixation of tissue samples by microwave irradiation for use in electron microscopy. Histochemical J. 25:719-725.  
         [0276]    51. Wessels, M. R. and M. S. Bronze. 1994. Critical role of the group A streptococcal capsule in pharyngeal colonization and infection in mice. Proc. Natl. Acad. Sci. USA 91:12238-12242.  
         [0277]    52. Wessels, M. R., A. E. Moses, J. B. Goldberg, and T. J. DiCesare. 1991. Hyaluronic acid capsule is a virulence factor for mucoid group A streptococci. Proc. Natl. Acad. Sci. USA. 88:8317-8321.  
         [0278]    53. Yamane, K., M. Kumamano, and K. Kurita. 1996. The 25°-36° region of the  Bacillus subtilis  chromosome: determination of the sequence of a 146 kb segment and identification of 113 genes. Microbiol. 142:3047-3056.  
         [0279]    54. Butler, J. C., R. F. Breiman, H. B. Lipman, J. Hofmann, and R. R. Facklam. 1995. Serotype distribution of  Streptococcus pneumoniae  infections among preschool children in the United States, 1978-1994: implications for development of a conjugate vaccine. J. Infect. Dis. 171:885-889.  
         [0280]    55. Charland, N., M. Jacques, S. Lacoutre and M. Gottschalk. 1997. Characterization and protective activity of a monoclonal antibody against a capsular epitope shared by  Streptococcus suis  serotypes 1, 2 and ½. Microbiol. 143:3607-3614.  
         [0281]    56. Gottschalk, M., R. Higgins, M. Jacques, K. R. Mittal, and J. Henrichsen. Description of 14 new capsular types of  Streptococcus suis.  J. Clin. Microbiol. 27:2633-2636.  
         [0282]    57. Heath, P. J., B. W. Hunt, and J. P. Duff. 1996.  Streptococcus suis  serotype 14 as a cause of pig disease in the UK. Vet. Rec. 2:450-451.  
         [0283]    58. Hommez, J., L. A. Devrieze, J. Henrichsen, and F. Castryck. 1986. Identification and characterization of  Streptococcus suis.  Vet. Microbiol. 16:349-355.  
         [0284]    59. Killper-Balz, R., and K. H. Schleifer. 1987. Streptococcus suis sp. nov. nom. rev. Int. J. Syst. Bacteriol. 37:160-162.  
         [0285]    60. Kolkman, M. A. B., B. A. M. van der Zeijst, and P. J. M. Nuijten. 1998. Diversity of capsular polysaccharide synthesis gene clusters in  Streptococcus pneumoniae.  Submitted for publication.  
         [0286]    61. Lee, J. C., S. Xu, A. Albus, and P. J. Livolsi. 1994. Genetic analysis of type 5 capsular polysaccharide expression by  Staphylococcus aureus.  J. Bacteriol. 176:4883-4889.  
         [0287]    62. Reek, F. H., M. A. Smits, E. M. Kamp, and H. E. Smith. 1995. Use of multiscreen plates for the preparation of bacterial DNA suitable for PCR. BioTechniques 19:282-285.  
         [0288]    63. Sau, S., N. Bhasin, E. R. Wann, J. C. Lee, T. J. Foster, and C. Y. Lee. 1997. The  Staphylococcus aureus  allelic genetic loci for serotype 5 and 8 capsule expression contain the type-specific genes flanked by common genes. Microbiol. 143:2395-2405.  
         [0289]    64. Sau, S., and C. Y. Lee. 1996. Cloning of type 8 capsule genes and analysis of gene clusters for the production of different capsular polysaccharides in  Staphylococcus aureus.  J. Bacteriol. 178:2118-2126.  
         [0290]    65. Sau, S., and C. Y. Lee. 1997. Molecular characterization and transcriptional analysis of type 8 capsule genes in  Staphylococcus aureus.  J. Bacteriol. 179:1614-1621.  
         [0291]    66. Smith, H. E., M. Rijnsburger, N. Stockhofe-Zurwieden, H. J. Wisselink, U. Vecht, and M. A. Smits. 1997. Virulent strains of  Streptococcus suis  serotype 2 and highly virulent strains of  Streptococcus suis  serotype 1 can be recognized by a unique ribotype profile. J. Clin. Microbiol. 35:1049-1053.  
         [0292]    67. Yamazaki, M., L. Thorne, M. Mikolajczak, R. W. Armentrout, and T. J. Pollock. 1996. Linkage of genes essential for synthesis of a polysaccharide capsule in  Sphingomonas  strain S88. J. Bacteriol. 178:2676-2687.  
         [0293]    68. Zhang, L., A. Al-Hendy, P. Toivanen. and M. Skuriik. 1993. Genetic organization and sequence of the  rfb  gene cluster of  Yersinia enterolitica  serotype 0:3: similarities to the dTDP-L-rhamnose biosynthesis pathway of  Salmonella  and to the bacterial polysaccharide transport systems. Mol. Microbiol. 9:309-321.  
         [0294]    69. Clifton-Hadley, F.A. (1983).  Streptococcus suis  type 2 infections. Br. Vet. J. 139, 1-5.  
         [0295]    70. Vecht, U., van Leengoed, L. A. M. G. and Verheyen, E. R. M. (1985).  Streptococcus suis  infections in pigs in The Netherlands (part one). Vet. Quart. 7, 315-321.  
         [0296]    71. Arends, J. P. and Zanen, H. C. (1988). Meningitis caused by  Streptococcus suis  in humans. Rev. Infect. Dis. 10, 131-137.  
         [0297]    72. Hommez, J., Devrieze, L.A., Henrichsen, J. and Castryck, F.(1986). Identification and characterization of  Streptococcus suis.  Vet. Microbiol. 16, 349-355.  
         [0298]    73. Killper-Balz, R. and Schleifer, K. H. (1987).  Streptococcus suis  sp. nov. nom.rev. Int. J. Syst. Bacteriol. 37, 160-162.  
         [0299]    74. Gottschalk, M., Higgins, R. and Jacques, M. (1993). Production of capsular material by  Streptococcus suis  serotype 2 under different conditions. Can. J. Vet. Res. 57, 49-52.  
         [0300]    75. Higgins, R. and Gottschalk, M. (1990). Un update on  Streptococcus suis  identification. J. Vet. Diagn. Invest. 2, 249-252.  
         [0301]    76. Gottschalk, M., Higgins, R., Jacques, M., Beaudoin, M. and Henrichsen, J. (1991). Characterization of six new capsular types (23 through 28) of  Streptococcus suis.  J. Clin. Microbiol. 29, 2590-2594.  
         [0302]    77. Gottschalk, M., Higgins, R., Jacques, M., Mittal, K. R. and Henrichsen, J. (1989) Description of 14 new capsular types of  Streptococcuss suis  J. Clin. Microbiol. 27, 2633-2636.  
         [0303]    78. Higgins, R., Gottschalk, M., Boudreau, M., Lebrun, A. and Henrichsen, J. (1995). Description of six new capsular types (28 through 34) of  Streptococcus suis.  J. Vet. Diagn. Invest. 7, 405-406.  
         [0304]    79. Aarestrup, F. M., Jorsal, S. E. and Jensen, N. E. (1998). Serological characterization and antimicrobial susceptibility of  Streptococcus suis  isolates from diagnostic samples in Denmark during 1995 and 1996. Vet. Microbiol. 15, 59-66.  
         [0305]    80. MacLennan, M., Foster, G., Dick, K., Smith, W. J. and Nielsen, B. (1996).  Streptococcus suis  serotypes 7, 8 and 14 from diseased pigs in Scotland. Vet Rec. 139, 423-424.  
         [0306]    81. Sihvonen, L., Kurl, D. N. and Henrichsen, J. (1988).  Streptococcus suis  isolates from pigs in Finland. Acta Vet. Scand. 29, 9-13.  
         [0307]    82. Boetner, A. G., Binder, M. and Bille-Hansen, V. (1987).  Streptococcus suis  infections in Danish pigs and experimental infection with Streptococcus suis serotype 7. Acta Path. Microbiol. Immunol. Scand. Sect. B, 95, 233-239.  
         [0308]    83. Smith, H. E., Veenbergen, V., van der Velde, J., Damman, M., Wisselink, H. J. and Smits, M. A. (1999). The cps genes of  Streptococcus suis  serotypes 1, 2 and 9: development of rapid serotype-specific PCR assays. J. Clin. Microbiol. submitted 84 Smith, H. E., Damman, M., van der Velde, J., Wagenaar, F., Wisselink, H. J., Stockhofe-Zurwieden, N. and Smits, M. A. (1999). Identification and characterization of the cps locus of  Streptococcus suis  serotype 2: the capsule protects against phagocytosis and is an important virulence factor. Infect. Immun. 67, 1750-1756.  
         [0309]    85. Miller, J. (1972). Experiments in Molecular Genetics. Cold Spring Harbor Laboratory, Cold Spring Harbor, N.Y.  
         [0310]    86. Sambrook, J., E. F. Fritsch, and T. Maniatis. (1989). Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, N.Y.  
         [0311]    87. Allen, A. and Maskell, D. (1996). The identification, cloning and mutagenesis of a genetic locus required for lipopolysaccharide biosynthesis in  Bordetella pertussis.  Mol. Microbiol. 19, 37-52.  
         [0312]    88. Wang, L. and Reeves, P. R. (1998). Organization of  Escherichia coli  O157 O antigen gene cluster and identification of its specific genes. Infect. Immun. 66, 3545-3551.  
         [0313]    89. Wisselink, H. J., Reek, F. H., Vecht, U., Stockhofe-Zurwieden, N., Smnits, M. A. and Smith, H. E. (1999). Detection of virulent strains of  Streptococcus suis  type 2 and highly, virulent strains of  Streptococcus suis  type 1 in tonsillar specimens of pigs by PCR. Vet. Microbiol. 67, 143-157.  
         [0314]    90. Konings, R. N. H., Verhoeven, E. J. M. and Peeters, B. P. H. (1987). pKUN vectors for the separate production of both DNA strands of recombinant plasmids. Methods Enzymol. 153, 12-34.  
     
       
       
         1 
         
           
             53  
           
           
             1  
             23  
             DNA  
             Artificial  
             
               Primer  
             
           
            1 

caaacgcaag gaattacggt atc                                             23 

 
           
             2  
             23  
             DNA  
             Artificial  
             
               primer  
             
           
            2 

gagtatctaa agaatgccta ttg                                             23 

 
           
             3  
             20  
             DNA  
             Artificial  
             
               primer  
             
           
            3 

ggcggtctag cagatgctcg                                                 20 

 
           
             4  
             19  
             DNA  
             Artificial  
             
               primer  
             
           
            4 

gcgaactgtt agcaatgac                                                  19 

 
           
             5  
             21  
             DNA  
             Artificial  
             
               primer  
             
           
            5 

ggctacatat aatggaagcc c                                               21 

 
           
             6  
             20  
             DNA  
             Artificial  
             
               primer  
             
           
            6 

cggaagtatc tgggctactg                                                 20 

 
           
             7  
             21  
             DNA  
             Artificial  
             
               primer  
             
           
            7 

agctctaaca cgaaataagg c                                               21 

 
           
             8  
             21  
             DNA  
             Artificial  
             
               primer  
             
           
            8 

gtcaaacacc ctggatagcc g                                               21 

 
           
             9  
             6992  
             DNA  
             Streptococcus suis  
             
               misc_feature  
               (1)..(6992)  
               CPS 2  
             
           
            9 

atcgccaaac gaaattggca ttatttgata tgatagcagt tgcaatttct gcaatcttaa     60 

caagtcatat accaaatgct gatttaaatc gttctggaat ttttatcata atgatggttc    120 

attattttgc attttttata tctcgtatgc cagttgaatt tgagtataga ggtaatctga    180 

tagagtttga aaaaacattt aactatagta taatatttgc aatttttctt acggcagtat    240 

catttttgtt ggagaataat ttcgcacttt caagacgtgg tgccgtgtat ttcacattaa    300 

taaacttcgt tttggtatac ctatttaacg taattattaa gcagtttaag gatagctttc    360 

tattttcgac aatctatcaa aaaaagacga ttctaattac aacggctgaa cgatgggaaa    420 

atatgcaagt tttatttgaa tcacataaac aaattcaaaa aaatcttgtt gcattggtag    480 

ttttaggtac agaaatagat aaaattaatt tatcattacc gctctattat tctgtggaag    540 

aagctataga gttttcaaca agggaagtgg tcgaccacgt ctttataaat ctaccaagtg    600 

agtttttaga cgtaaagcaa ttcgtttcag attttgagtt gttaggtatt gatgtaagcg    660 

ttgatattaa ttcattcggt tttactgcgt tgaaaaacaa aaaaatccaa ctgctaggtg    720 

accatagcat tgtaactttt tccacaaatt tttataagcc tagtcatatc atgatgaaac    780 

gacttttgga tatactcgga gcggtagtcg ggttaattat ttgtggtata gtttctattt    840 

tgttagttcc aattattcgt agagatggtg gaccggctat ttttgctcag aaacgagttg    900 

gacagaatgg acgcatattt acattctaca agtttcgatc gatgtatgtt gatgctgagg    960 

agcgcaaaaa agacttgctc agccaaaacc agatgcaagg gtgggtatgt tttaaaatgg   1020 

gaaaaacgat cctagaatta ctccaattgg acatttcata cgcaaaaaca agtttagacg   1080 

agttaccaca gttttataat gttttaattg gcgatatgag tctagttggt acacgtccac   1140 

ctacagttga tgaatttgaa aaatatactc ctggtcaaaa gagacgattg agttttaaac   1200 

cagggattac aggtctctgg caggttagtg gtcgtagtaa tatcacagac ttcgacgacg   1260 

tagttcggtt ggacttagca tacattgata attggactat ctggtcagat attaaaattt   1320 

tattaaagac agtgaaagtt gtattgttga gagagggaag taagtaaaag tatatgaaag   1380 

tttgtttggt cggttcttca gggggacatt tgactcactt gtatttgtta aaaccgtttt   1440 

ggaaggaaga agaacgtttt tgggtaacat ttgataaaga ggatgcaaga agtcttttga   1500 

agaatgaaaa aatgtatcca tgttactttc caacaaatcg caatctcatt aatttagtga   1560 

aaaatacttt cttagctttc aaaattttac gtgatgagaa accagatgtt attatttcat   1620 

ctggtgcggc cgttgctgtc cccttctttt acatcggaaa actatttgga gcaaagacga   1680 

tttatattga agtatttgat cgagttaata aatctacatt aactggaaaa ctagtttatc   1740 

ccgtaacaga tatttttatt gttcagtggg aagaaatgaa gaaggtatat cctaaatcta   1800 

ttaacttggg gagtattttt taatgatttt tgtaacagta ggaactcatg aacaacagtt   1860 

taatcgattg ataaaagaga ttgatttatt gaaaaaaaat ggaagtataa ccgacgaaat   1920 

atttattcaa acaggatatt ctgactatat tccagaatat tgcaagtata aaaaatttct   1980 

cagttacaaa gaaatggaac aatatattaa caaatcagaa gtagttattt gccacggagg   2040 

ccccgctact tttatgaatt cattatccaa aggaaaaaaa caattattgt ttcctagaca   2100 

aaaaaagtat ggtgaacatg taaatgatca tcaagtagag tttgtaagaa gaattttaca   2160 

agataataat attttattta tagaaaatat agatgatttg tttgaaaaaa ttattgaagt   2220 

ttctaagcaa actaacttta catcaaataa taattttttt tgtgaaagat taaaacaaat   2280 

agttgaaaaa tttaatgagg atcaagaaaa tgaataataa aaaagatgca tatttgataa   2340 

tggcttatca taatttttct cagattttac tggagaggga tacagatatt atcatcttct   2400 

ctcaggagaa tgcacaccat tagttccttc agaatacctg tataattatt ttaaatattc   2460 

tcaggattta tatgttgaat ttacaaaaga tgagcaaaaa tataaagaaa ataggatata   2520 

tgaacgagtt aaatgttaca gattatttcc taatatatca gaaaaaacta ttgataatgt   2580 

actgtttaga attttattaa gaatgtatcg agcttttgaa tactatttac aaagattgtt   2640 

gtttattgat agaataaaaa acatggtcta agaataagat ttggttctaa ttgggtttcg   2700 

cttccacatg attttgtggc aattctttta tcaaatgaaa acgaaacagc ttatttattt   2760 

aagtaatcta aatgtccaga tgaactattt atacagacaa ttatagaaaa atatgaattt   2820 

tcaaatagat tatctaaata tggaaattta agatatataa agtggaaaaa atcaacatct   2880 

tctcctattg tctttacaga tgattctatt gatgaattgc taaatgcaag aaatttaggt   2940 

tttttatttg ctagaaagtt aaaaatagaa aataaatcta aatttaaaga aattattact   3000 

aaaaaataaa atagttgatt ttgtgagagt aatgtatgtt taaattattt aaatatgacc   3060 

cggaatattt tatttttaag tacttctggt tgattatttt tattccagag caaaagtatg   3120 

tatttttatt aatttttatg aatttaattt tatttcatat aaaatttttg aaaactaagc   3180 

taatattaaa aaatgaaatt ttattgtttt tattatggtc tatattatgt tttgtttcag   3240 

tagtcacaag tatgtttgtt gaaataaatt ttgaaagatt atttgcagat tttactgctc   3300 

ccataatttg gattattgca ataatgtatt ataatttgta ttcatttata aatattgatt   3360 

ataaaaaatt aaaaaatagt atctttttta gttttttagt tttattaggt atatctgcat   3420 

tgtatattat tcaaaatggg aaagatattg tatttttaga cagacacctt ataggactag   3480 

actatcttat aacaggcgtc aaaacaaggt tggttggctt tatgaactat cctacgttaa   3540 

ataccactac aattatagtt tcaattccgt taatctttgc acttataaaa aataaaatgc   3600 

aacaattttt tttcttgtgt cttgctttta taccgatcta tttaagtgga tcgagaattg   3660 

gtagtttatc gctagcaata ttaattatat gcttgttatg gagatatata ggtggaaaat   3720 

ttgcttggat aaaaaagcta atagtaatat ttgtaatact acttattatt ttaaatactg   3780 

aattgcttta ccatgaaatt ttggctgttt ataattctag agaatcaagt aacgaagcta   3840 

gatttattat ttatcaagga agtattgata aagtattaga aaacaatatt ttatttggat   3900 

atggaatatc cgaatattca gttacgggaa cttggctcgg aagtcattca ggctatatat   3960 

cattttttta taaatcagga atagttgggt tgattttact gatgttttct tttttttatg   4020 

ttataaaaaa aagttatgga gttaatgggg aaacagcact attttatttt acatcattag   4080 

ccatattttt catatatgaa acaatagatc cgattattat tatattagta ctattctttt   4140 

cttcaatagg tatttggaat aatataaatt ttaaaaagga tatggagaca aaaaatgaat   4200 

gatttaattt cagttattgt accaatttat aatgtccaag attatcttga taaatgtatt   4260 

aacagtatta ttaaccaaac atatactaat ttagaggtta ttctcgtaaa tgatggaagt   4320 

actgatgatt ctgagaaaat ttgcttaaac tatatgaaga acgatggaag aattaaatat   4380 

tacaagaaaa ttaatggcgg tctagcagat gctcgaaatt tcggactaga acatgcaaca   4440 

ggtaaatata ttgcttttgt cgattctgat gactatatag aagttgcaat gttcgagaga   4500 

atgcatgata atataactga gtataatgcc gatatagcag agatagattt ttgtttagta   4560 

gacgaaaacg ggtatacaaa gaaaaaaaga aatagtaatt ttcatgtctt aacgagagaa   4620 

gagactgtaa aagaattttt gtcaggatct aatatagaaa ataatgtttg gtgcaagctt   4680 

tattcacgag atattataaa agatataaaa ttccaaatta ataatagaag tattggtgag   4740 

gatttgcttt ttaatttgga ggtcttgaac aatgtaacac gtgtagtagt tgatactaga   4800 

gaatattatt ataattatgt cattcgtaac agttcgctta ttaatcagaa attctctata   4860 

aataatattg atttagtcac aagattggag aattacccct ttaagttaaa aagagagttt   4920 

agtcattatt ttgatgcaaa agttattaaa gagaaggtta aatgtttaaa caaaatgtat   4980 

tcaacagatt gtttggataa tgagttcttg ccaatattag agtcttatcg aaaagaaata   5040 

cgtagatatc catttattaa agcgaaaaga tatttatcaa gaaagcattt agttacgttg   5100 

tatttgatga aattttcgcc taaactatat gtaatgttat ataagaaatt tcaaaagcag   5160 

tagaggtaaa aatggataaa attagtgtta ttgttccagt ttataatgta gataaatatt   5220 

taagtagttg tatagaaagc attattaatc aaaattataa aaatatagaa atattattga   5280 

tagatgatgg ctctgtagat gattctgcta aaatatgcaa ggaatatgca gaaaaagata   5340 

aaagagtaaa aatttttttc actaatcata gtggagtatc aaatgctaga aatcatggaa   5400 

taaagcggag tacagctgaa tatattatgt ttgttgactc tgatgatgtt gttgatagta   5460 

gattagtaga aaaattatat tttaatatta taaaaagtag aagtgattta tctggttgtt   5520 

tgtacgctac tttttcagaa aatataaata attttgaagt gaataatcca aatattgatt   5580 

ttgaagcaat taataccgtg caggacatgg gagaaaaaaa ttttatgaat ttgtatataa   5640 

ataatatttt ttctactcct gtttgtaaac tatataagaa aagatacata acagatcttt   5700 

ttcaagagaa tcaatggtta ggagaagatt tactttttaa tctgcattat ttaaagaata   5760 

tagatagagt tagttatttg actgaacatc tttattttta taggagaggt atactaagta   5820 

cagtaaattc ttttaaagaa ggtgtgtttt tgcaattgga aaatttgcaa aaacaagtga   5880 

tagtattgtt taagcaaata tatggtgagg attttgacgt atcaattgtt aaagatacta   5940 

tacgttggca agtattttat tatagcttac taatgtttaa atacggaaaa cagtctattt   6000 

ttgacaaatt tttaattttt agaaatcttt ataaaaaata ttattttaac ttgttaaaag   6060 

tatctaacaa aaattctttg tctaaaaatt tttgtataag aattgtttcg aacaaagttt   6120 

ttaaaaaaat attatggtta taataggaag atatcatgga tactattagt aaaatttcta   6180 

taattgtacc tatatataat gtagaaaaat atttatctaa atgtatagat agcattgtaa   6240 

atcagaccta caaacatata gagattcttc tggtgaatga cggtagtacg gataattcgg   6300 

aagaaatttg tttagcatat gcgaagaaag atagtcgcat tcgttatttt aaaaaagaga   6360 

acggcgggct atcagatgcc cgtaattatg gcataagtcg cgccaagggt gactacttag   6420 

cttttataga ctcagatgat tttattcatt cggagttcat ccaacgttta cacgaagcaa   6480 

ttgagagaga gaatgccctt gtggcagttg ctggttatga tagggtagat gcttcggggc   6540 

atttcttaac agcagagccg cttcctacaa atcaggctgt tctgagcggc aggaatgttt   6600 

gtaaaaagct gctagaggcg gatggtcatc gctttgtggt ggcctgtaat aaactctata   6660 

aaaaagaact atttgaagat tttcgatttg aaaagggtaa gattcatgaa gatgaatact   6720 

tcacttatcg cttgctctat gagttagaaa aagttgcaat agttaaggag tgcttgtact   6780 

attatgttga ccgagaaaat agtatcacaa cttctagcat gactgaccat cgcttccatt   6840 

gcctactgga atttcaaaat gaacgaatgg acttctatga aagtagagga gataaagagc   6900 

tcttactaga gtgttatcgt tcatttttag cctttgctgt tttgttttta ggcaaatata   6960 

atcattggtt gagcaaacag caaaagaagc tt                                 6992 

 
           
             10  
             239  
             PRT  
             Streptococcus suis  
             
               misc_feature  
               ORF2Z  
             
           
            10 

Ser Leu Asp Ile Asp His Met Met Glu Val Met Glu Ala Ser Lys Ser 
1               5                   10                  15 

Ala Ala Gly Ser Ala Cys Pro Ser Pro Gln Ala Tyr Gln Ala Ala Phe 
            20                  25                  30 

Glu Gly Ala Glu Asn Ile Ile Val Val Thr Ile Thr Gly Gly Leu Ser 
        35                  40                  45 

Gly Ser Phe Asn Ala Ala Arg Val Ala Arg Asp Met Tyr Ile Glu Glu 
    50                  55                  60 

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

Glu Met Asp Leu Leu Val His Gln Ile Asn Arg Leu Ile Ser Ala Gly 
                85                  90                  95 

Leu Asp Phe Pro Gln Val Val Glu Ala Ile Thr His Tyr Arg Glu His 
            100                 105                 110 

Ser Lys Leu Leu Phe Val Leu Ala Lys Val Asp Asn Leu Val Lys Asn 
        115                 120                 125 

Gly Arg Leu Ser Lys Leu Val Gly Thr Val Val Gly Leu Leu Asn Ile 
    130                 135                 140 

Arg Met Val Gly Glu Ala Ser Ala Glu Gly Lys Leu Glu Leu Leu Gln 
145                 150                 155                 160 

Lys Ala Arg Gly His Lys Lys Ser Val Thr Ala Ala Phe Glu Glu Met 
                165                 170                 175 

Lys Lys Ala Gly Tyr Asp Gly Gly Arg Ile Val Met Ala His Arg Asn 
            180                 185                 190 

Asn Ala Lys Phe Phe Gln Gln Phe Ser Glu Leu Val Lys Ala Ser Phe 
        195                 200                 205 

Pro Thr Ala Val Ile Asp Glu Val Ala Thr Ser Gly Leu Cys Ser Phe 
    210                 215                 220 

Tyr Ala Glu Glu Gly Gly Leu Leu Met Gly Tyr Glu Val Lys Ala 
225                 230                 235 

 
           
             11  
             244  
             PRT  
             Streptococcus suis  
             
               misc_feature  
               ORF2X  
             
           
            11 

Met Lys Ile Ile Ile Pro Asn Ala Lys Glu Val Asn Thr Asn Leu Glu 
1               5                   10                  15 

Asn Ala Ser Phe Tyr Leu Leu Ser Asp Arg Ser Lys Pro Val Leu Asp 
            20                  25                  30 

Ala Ile Ser Gln Phe Asp Val Lys Lys Met Ala Ala Phe Tyr Lys Leu 
        35                  40                  45 

Asn Glu Ala Lys Ala Glu Leu Glu Ala Asp Arg Trp Tyr Arg Ile Arg 
    50                  55                  60 

Thr Gly Gln Ala Lys Thr Tyr Pro Ala Trp Gln Leu Tyr Asp Gly Leu 
65                  70                  75                  80 

Met Tyr Arg Tyr Met Asp Arg Arg Gly Ile Asp Ser Lys Glu Glu Asn 
                85                  90                  95 

Tyr Leu Arg Asp His Val Arg Val Ala Thr Ala Leu Tyr Gly Leu Ile 
            100                 105                 110 

His Pro Phe Glu Phe Ile Ser Pro His Arg Leu Asp Phe Gln Gly Ser 
        115                 120                 125 

Leu Lys Ile Gly Asn Gln Ser Leu Lys Gln Tyr Trp Arg Pro Tyr Tyr 
    130                 135                 140 

Asp Gln Glu Val Gly Asp Asp Glu Leu Ile Leu Ser Leu Ala Ser Ser 
145                 150                 155                 160 

Glu Phe Glu Gln Val Phe Ser Pro Gln Ile Gln Lys Arg Leu Val Lys 
                165                 170                 175 

Ile Leu Phe Met Glu Glu Lys Ala Gly Gln Leu Lys Val His Ser Thr 
            180                 185                 190 

Ile Ser Lys Lys Gly Arg Gly Arg Leu Leu Ser Trp Leu Ala Lys Asn 
        195                 200                 205 

Asn Ile Gln Glu Leu Ser Asp Ile Gln Asp Phe Lys Val Asp Gly Phe 
    210                 215                 220 

Glu Tyr Cys Thr Ser Glu Ser Thr Ala Asn Gln Leu Thr Phe Ile Arg 
225                 230                 235                 240 

Ser Ile Lys Met 

 
           
             12  
             481  
             PRT  
             Streptococcus suis  
             
               misc_feature  
               CPS2A  
             
           
            12 

Met Lys Lys Arg Ser Gly Arg Ser Lys Ser Ser Lys Phe Lys Leu Val 
1               5                   10                  15 

Asn Phe Ala Leu Leu Gly Leu Tyr Ser Ile Thr Leu Cys Leu Phe Leu 
            20                  25                  30 

Val Thr Met Tyr Arg Tyr Asn Ile Leu Asp Phe Arg Tyr Leu Asn Tyr 
        35                  40                  45 

Ile Val Thr Leu Leu Leu Val Gly Val Ala Val Leu Ala Gly Leu Leu 
    50                  55                  60 

Met Trp Arg Lys Lys Ala Arg Ile Phe Thr Ala Leu Leu Leu Val Phe 
65                  70                  75                  80 

Ser Leu Val Ile Thr Ser Val Gly Ile Tyr Gly Met Gln Glu Val Val 
                85                  90                  95 

Lys Phe Ser Thr Arg Leu Asn Ser Asn Ser Thr Phe Ser Glu Tyr Glu 
            100                 105                 110 

Met Ser Ile Leu Val Pro Ala Asn Ser Asp Ile Thr Asp Val Arg Gln 
        115                 120                 125 

Leu Thr Ser Ile Leu Ala Pro Ala Glu Tyr Asp Gln Asp Asn Ile Thr 
    130                 135                 140 

Ala Leu Leu Asp Asp Ile Ser Lys Met Glu Ser Thr Gln Leu Ala Thr 
145                 150                 155                 160 

Ser Pro Gly Thr Ser Tyr Leu Thr Ala Tyr Gln Ser Met Leu Asn Gly 
                165                 170                 175 

Glu Ser Gln Ala Met Val Phe Asn Gly Val Phe Thr Asn Ile Leu Glu 
            180                 185                 190 

Asn Glu Asp Pro Gly Phe Ser Ser Lys Val Lys Lys Ile Tyr Ser Phe 
        195                 200                 205 

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

Ser Phe Asn Ile Tyr Ile Ser Gly Ile Asp Ala Tyr Gly Pro Ile Ser 
225                 230                 235                 240 

Thr Val Ser Arg Ser Asp Val Asn Ile Ile Met Thr Val Asn Arg Ala 
                245                 250                 255 

Thr His Lys Ile Leu Leu Thr Thr Thr Pro Arg Asp Ser Tyr Val Ala 
            260                 265                 270 

Phe Ala Asp Gly Gly Gln Asn Gln Tyr Asp Lys Leu Thr His Ala Gly 
        275                 280                 285 

Ile Tyr Gly Val Asn Ala Ser Val His Thr Leu Glu Asn Phe Tyr Gly 
    290                 295                 300 

Ile Asp Ile Ser Asn Tyr Val Arg Leu Asn Phe Ile Ser Phe Leu Gln 
305                 310                 315                 320 

Leu Ile Asp Leu Val Gly Gly Ile Asp Val Tyr Asn Asp Gln Glu Phe 
                325                 330                 335 

Thr Ser Leu His Gly Asn Tyr His Phe Pro Val Gly Gln Val His Leu 
            340                 345                 350 

Asn Ser Asp Gln Ala Leu Gly Phe Val Arg Glu Arg Tyr Ser Leu Thr 
        355                 360                 365 

Gly Gly Asp Asn Asp Arg Gly Lys Asn Gln Glu Lys Val Ile Ala Ala 
    370                 375                 380 

Leu Ile Lys Lys Met Ser Thr Pro Glu Asn Leu Lys Asn Tyr Gln Ala 
385                 390                 395                 400 

Ile Leu Ser Gly Leu Glu Gly Ser Ile Gln Thr Asp Leu Ser Leu Glu 
                405                 410                 415 

Thr Ile Met Ser Leu Val Asn Thr Gln Leu Glu Ser Gly Thr Gln Phe 
            420                 425                 430 

Thr Val Glu Ser Gln Ala Leu Thr Gly Thr Gly Arg Ser Asp Leu Ser 
        435                 440                 445 

Ser Tyr Ala Met Pro Gly Ser Gln Leu Tyr Met Met Glu Ile Asn Gln 
    450                 455                 460 

Asp Ser Leu Glu Gln Ser Lys Ala Ala Ile Gln Ser Val Leu Val Glu 
465                 470                 475                 480 

Lys 

 
           
             13  
             229  
             PRT  
             Streptococcus suis  
             
               misc_feature  
               CPS2B  
             
           
            13 

Met Asn Asn Gln Glu Val Asn Ala Ile Glu Ile Asp Val Leu Phe Leu 
1               5                   10                  15 

Leu Lys Thr Ile Trp Arg Lys Lys Phe Leu Ile Leu Leu Thr Ala Val 
            20                  25                  30 

Leu Thr Ala Gly Leu Ala Phe Val Tyr Ser Ser Phe Leu Val Thr Pro 
        35                  40                  45 

Gln Tyr Asp Ser Thr Thr Arg Ile Tyr Val Val Ser Gln Asn Val Glu 
    50                  55                  60 

Ala Gly Ala Gly Leu Thr Asn Gln Glu Leu Gln Ala Gly Thr Tyr Leu 
65                  70                  75                  80 

Ala Lys Asp Tyr Arg Glu Ile Ile Leu Ser Gln Asp Val Leu Thr Gln 
                85                  90                  95 

Val Ala Thr Glu Leu Asn Leu Lys Glu Ser Leu Lys Glu Lys Ile Ser 
            100                 105                 110 

Val Ser Ile Pro Val Asp Thr Arg Ile Val Ser Ile Ser Val Arg Asp 
        115                 120                 125 

Ala Asp Pro Asn Glu Ala Ala Arg Ile Ala Asn Ser Leu Arg Thr Phe 
    130                 135                 140 

Ala Val Gln Lys Val Val Glu Val Thr Lys Val Ser Asp Val Thr Thr 
145                 150                 155                 160 

Leu Glu Glu Ala Val Pro Ala Glu Glu Pro Thr Thr Pro Asn Thr Lys 
                165                 170                 175 

Arg Asn Ile Leu Leu Gly Leu Leu Ala Gly Gly Ile Leu Ala Thr Gly 
            180                 185                 190 

Leu Val Leu Val Met Glu Val Leu Asp Asp Arg Val Lys Arg Pro Gln 
        195                 200                 205 

Asp Ile Glu Glu Val Met Gly Leu Thr Leu Leu Gly Ile Val Pro Asp 
    210                 215                 220 

Ser Lys Lys Leu Lys 
225 

 
           
             14  
             225  
             PRT  
             Streptococcus suis  
             
               misc_feature  
               CPS2C  
             
           
            14 

Met Ala Met Leu Glu Ile Ala Arg Thr Lys Arg Glu Gly Val Asn Lys 
1               5                   10                  15 

Thr Glu Glu Tyr Phe Asn Ala Ile Arg Thr Asn Ile Gln Leu Ser Gly 
            20                  25                  30 

Ala Asp Ile Lys Val Val Gly Ile Thr Ser Val Lys Ser Asn Glu Gly 
        35                  40                  45 

Lys Ser Thr Thr Ala Ala Ser Leu Ala Ile Ala Tyr Ala Arg Ser Gly 
    50                  55                  60 

Tyr Lys Thr Val Leu Val Asp Ala Asp Ile Arg Asn Ser Val Met Pro 
65                  70                  75                  80 

Gly Phe Phe Lys Pro Ile Thr Lys Ile Thr Gly Leu Thr Asp Tyr Leu 
                85                  90                  95 

Ala Gly Thr Thr Asp Leu Ser Gln Gly Leu Cys Asp Thr Asp Ile Pro 
            100                 105                 110 

Asn Leu Thr Val Ile Glu Ser Gly Lys Val Ser Pro Asn Pro Thr Ala 
        115                 120                 125 

Leu Leu Gln Ser Lys Asn Phe Glu Asn Leu Leu Ala Thr Leu Arg Arg 
    130                 135                 140 

Tyr Tyr Asp Tyr Val Ile Val Asp Cys Pro Pro Leu Gly Leu Val Ile 
145                 150                 155                 160 

Asp Ala Ala Ile Ile Ala Gln Lys Cys Asp Ala Met Val Ala Val Val 
                165                 170                 175 

Glu Ala Gly Asn Val Lys Cys Ser Ser Leu Lys Lys Val Lys Glu Gln 
            180                 185                 190 

Leu Glu Gln Thr Gly Thr Pro Phe Leu Gly Val Ile Leu Asn Lys Tyr 
        195                 200                 205 

Asp Ile Ala Thr Glu Lys Tyr Ser Glu Tyr Gly Asn Tyr Gly Lys Lys 
    210                 215                 220 

Ala 
225 

 
           
             15  
             243  
             PRT  
             Streptococcus suis  
             
               misc_feature  
               CPS2D  
             
           
            15 

Met Ile Asp Ile His Ser His Ile Ile Phe Gly Val Asp Asp Gly Pro 
1               5                   10                  15 

Lys Thr Ile Glu Glu Ser Leu Ser Leu Ile Ser Glu Ala Tyr Arg Gln 
            20                  25                  30 

Gly Val Arg Tyr Ile Val Ala Thr Ser His Arg Arg Lys Gly Met Phe 
        35                  40                  45 

Glu Thr Pro Glu Lys Ile Ile Met Ile Asn Phe Leu Gln Leu Lys Glu 
    50                  55                  60 

Ala Val Ala Glu Val Tyr Pro Glu Ile Arg Leu Cys Tyr Gly Ala Glu 
65                  70                  75                  80 

Leu Tyr Tyr Ser Lys Asp Ile Leu Ser Lys Leu Glu Lys Lys Lys Val 
                85                  90                  95 

Pro Thr Leu Asn Gly Ser Cys Tyr Ile Leu Leu Glu Phe Ser Thr Asp 
            100                 105                 110 

Thr Pro Trp Lys Glu Ile Gln Glu Ala Val Asn Glu Met Thr Leu Leu 
        115                 120                 125 

Gly Leu Thr Pro Val Leu Ala His Ile Glu Arg Tyr Asp Ala Leu Ala 
    130                 135                 140 

Phe Gln Ser Glu Arg Val Glu Lys Leu Ile Asp Lys Gly Cys Tyr Thr 
145                 150                 155                 160 

Gln Val Asn Ser Asn His Val Leu Lys Pro Ala Leu Ile Gly Glu Arg 
                165                 170                 175 

Ala Lys Glu Phe Lys Lys Arg Thr Arg Tyr Phe Leu Glu Gln Asp Leu 
            180                 185                 190 

Val His Cys Val Ala Ser Asp Met His Asn Leu Tyr Ser Arg Pro Pro 
        195                 200                 205 

Phe Met Arg Glu Ala Tyr Gln Leu Val Lys Lys Glu Tyr Gly Glu Asp 
    210                 215                 220 

Arg Ala Lys Ala Leu Phe Lys Lys Asn Pro Leu Leu Ile Leu Lys Asn 
225                 230                 235                 240 

Gln Val Gln 

 
           
             16  
             459  
             PRT  
             Streptococcus suis  
             
               misc_feature  
               CPS2E  
             
           
            16 

Met Asn Ile Glu Ile Gly Tyr Arg Gln Thr Lys Leu Ala Leu Phe Asp 
1               5                   10                  15 

Met Ile Ala Val Thr Ile Ser Ala Ile Leu Thr Ser His Ile Pro Asn 
            20                  25                  30 

Ala Asp Leu Asn Arg Ser Gly Ile Phe Ile Ile Met Met Val His Tyr 
        35                  40                  45 

Phe Ala Phe Phe Ile Ser Arg Met Pro Val Glu Phe Glu Tyr Arg Gly 
    50                  55                  60 

Asn Leu Ile Glu Phe Glu Lys Thr Phe Asn Tyr Ser Ile Ile Phe Val 
65                  70                  75                  80 

Ile Phe Leu Met Ala Val Ser Phe Met Leu Glu Asn Asn Phe Ala Leu 
                85                  90                  95 

Ser Arg Arg Gly Ala Val Tyr Phe Thr Leu Ile Asn Phe Val Leu Val 
            100                 105                 110 

Tyr Leu Phe Asn Val Ile Ile Lys Gln Phe Lys Asp Ser Phe Leu Phe 
        115                 120                 125 

Ser Thr Thr Tyr Gln Lys Lys Thr Ile Leu Ile Thr Thr Ala Glu Leu 
    130                 135                 140 

Trp Glu Asn Met Gln Val Leu Phe Glu Ser Asp Ile Leu Phe Gln Lys 
145                 150                 155                 160 

Asn Leu Val Ala Leu Val Ile Leu Gly Thr Glu Ile Asp Lys Ile Asn 
                165                 170                 175 

Leu Pro Leu Pro Leu Tyr Tyr Ser Val Glu Glu Ala Ile Gly Phe Ser 
            180                 185                 190 

Thr Arg Glu Val Val Asp Tyr Val Phe Ile Asn Leu Pro Ser Glu Tyr 
        195                 200                 205 

Phe Asp Leu Lys Gln Leu Val Ser Asp Phe Glu Leu Leu Gly Ile Asp 
    210                 215                 220 

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

Lys Ile Gln Met Leu Gly Asp His Ser Ile Val Thr Phe Ser Thr Asn 
                245                 250                 255 

Phe Tyr Lys Pro Ser His Ile Trp Met Lys Arg Leu Leu Asp Ile Leu 
            260                 265                 270 

Gly Ala Val Val Gly Leu Ile Ile Ser Gly Ile Val Ser Ile Leu Leu 
        275                 280                 285 

Ile Pro Ile Ile Arg Arg Asp Gly Gly Pro Ala Ile Phe Ala Gln Lys 
    290                 295                 300 

Arg Val Gly Gln Asn Gly Arg Ile Phe Thr Phe Tyr Lys Phe Arg Ser 
305                 310                 315                 320 

Met Phe Val Asp Ala Glu Val Arg Lys Lys Glu Leu Met Ala Gln Asn 
                325                 330                 335 

Gln Met Gln Gly Gly Met Phe Lys Met Asp Asn Asp Pro Arg Ile Thr 
            340                 345                 350 

Pro Ile Gly His Phe Ile Arg Lys Thr Ser Leu Asp Glu Leu Pro Gln 
        355                 360                 365 

Phe Tyr Asn Val Leu Ile Gly Asp Met Ser Leu Val Gly Thr Arg Pro 
    370                 375                 380 

Pro Thr Val Asp Glu Phe Glu Lys Tyr Thr Pro Ser Gln Lys Arg Arg 
385                 390                 395                 400 

Leu Ser Phe Lys Pro Gly Ile Thr Gly Leu Trp Gln Val Ser Gly Arg 
                405                 410                 415 

Ser Asp Ile Thr Asp Phe Asn Glu Val Val Arg Leu Asp Leu Thr Tyr 
            420                 425                 430 

Ile Asp Asn Trp Thr Ile Trp Ser Asp Ile Lys Ile Leu Leu Lys Thr 
        435                 440                 445 

Val Lys Val Val Leu Leu Arg Glu Gly Gly Gln 
    450                 455 

 
           
             17  
             389  
             PRT  
             Streptococcus suis  
             
               misc_feature  
               CPS2F  
             
           
            17 

Met Arg Thr Val Tyr Ile Ile Gly Ser Lys Gly Ile Pro Ala Lys Tyr 
1               5                   10                  15 

Gly Gly Phe Glu Thr Phe Val Glu Lys Leu Thr Glu Tyr Gln Lys Asp 
            20                  25                  30 

Lys Ser Ile Asn Tyr Phe Val Ala Cys Thr Arg Glu Asn Ser Ala Lys 
        35                  40                  45 

Ser Asp Ile Thr Gly Glu Val Phe Glu His Asn Gly Ala Thr Cys Phe 
    50                  55                  60 

Asn Ile Asp Val Pro Asn Ile Gly Ser Ala Lys Ala Ile Leu Tyr Asp 
65                  70                  75                  80 

Ile Met Ala Leu Lys Lys Ser Ile Glu Ile Ala Lys Asp Arg Asn Asp 
                85                  90                  95 

Thr Ser Pro Ile Phe Tyr Ile Leu Ala Cys Arg Ile Gly Pro Phe Ile 
            100                 105                 110 

Tyr Leu Phe Lys Lys Gln Ile Glu Ser Ile Gly Gly Gln Leu Phe Val 
        115                 120                 125 

Asn Pro Asp Gly His Glu Trp Leu Arg Glu Lys Trp Ser Tyr Pro Val 
    130                 135                 140 

Arg Gln Tyr Trp Lys Phe Ser Glu Ser Leu Met Leu Lys Tyr Ala Asp 
145                 150                 155                 160 

Leu Leu Ile Cys Asp Ser Lys Asn Ile Glu Lys Tyr Ile His Glu Asp 
                165                 170                 175 

Tyr Arg Lys Tyr Ala Pro Glu Thr Ser Tyr Ile Ala Tyr Gly Thr Asp 
            180                 185                 190 

Leu Asp Lys Ser Arg Leu Ser Pro Thr Asp Ser Val Val Arg Glu Trp 
        195                 200                 205 

Tyr Lys Glu Lys Glu Ile Ser Glu Asn Asp Tyr Tyr Leu Val Val Gly 
    210                 215                 220 

Arg Phe Val Pro Glu Asn Asn Tyr Glu Val Met Ile Arg Glu Phe Met 
225                 230                 235                 240 

Lys Ser Tyr Ser Arg Lys Asp Phe Val Leu Ile Thr Asn Val Glu His 
                245                 250                 255 

Asn Ser Phe Tyr Glu Lys Leu Lys Lys Glu Thr Gly Phe Asp Lys Asp 
            260                 265                 270 

Lys Arg Ile Lys Phe Val Gly Thr Val Tyr Asn Gln Glu Leu Leu Lys 
        275                 280                 285 

Tyr Ile Arg Glu Asn Ala Phe Ala Tyr Phe His Gly His Glu Val Gly 
    290                 295                 300 

Gly Thr Asn Pro Ser Leu Leu Glu Ala Leu Ser Ser Thr Lys Leu Asn 
305                 310                 315                 320 

Leu Leu Leu Asp Val Gly Phe Asn Arg Glu Val Gly Glu Glu Gly Ala 
                325                 330                 335 

Lys Tyr Trp Asn Lys Asp Asn Leu His Arg Val Ile Asp Ser Cys Glu 
            340                 345                 350 

Gln Leu Ser Gln Glu Gln Ile Asn Asp Met Asp Ser Leu Ser Thr Lys 
        355                 360                 365 

Gln Val Lys Glu Arg Phe Ser Trp Asp Phe Ile Val Asp Glu Tyr Glu 
    370                 375                 380 

Lys Leu Phe Lys Gly 
385 

 
           
             18  
             385  
             PRT  
             Streptococcus suis  
             
               misc_feature  
               CPS2G  
             
           
            18 

Met Lys Lys Ile Leu Tyr Leu His Ala Gly Ala Glu Leu Tyr Gly Ala 
1               5                   10                  15 

Asp Lys Val Leu Leu Glu Leu Ile Lys Gly Leu Asp Lys Asn Glu Phe 
            20                  25                  30 

Glu Ala His Val Ile Leu Pro Asn Asp Gly Val Leu Val Pro Ala Leu 
        35                  40                  45 

Arg Glu Val Gly Ala Gln Val Glu Val Ile Asn Tyr Pro Ile Leu Arg 
    50                  55                  60 

Arg Lys Tyr Phe Asn Pro Lys Gly Ile Phe Asp Tyr Phe Ile Ser Tyr 
65                  70                  75                  80 

His His Tyr Ser Lys Gln Ile Ala Gln Tyr Ala Ile Glu Asn Lys Val 
                85                  90                  95 

Asp Ile Ile His Asn Asn Thr Thr Ala Val Leu Glu Gly Ile Tyr Leu 
            100                 105                 110 

Lys Arg Lys Leu Lys Leu Pro Leu Leu Trp His Val His Glu Ile Ile 
        115                 120                 125 

Val Lys Pro Lys Phe Ile Ser Asp Ser Ile Asn Phe Leu Met Gly Arg 
    130                 135                 140 

Phe Ala Asp Lys Ile Val Thr Val Ser Gln Ala Val Ala Asn His Ile 
145                 150                 155                 160 

Lys Gln Ser Pro His Ile Lys Asp Asp Gln Ile Ser Val Ile Tyr Asn 
                165                 170                 175 

Gly Val Asp Asn Lys Val Phe Tyr Gln Ser Asp Ala Arg Ser Val Arg 
            180                 185                 190 

Glu Arg Phe Asp Ile Asp Glu Glu Ala Leu Val Ile Gly Met Val Gly 
        195                 200                 205 

Arg Val Asn Ala Trp Lys Gly Gln Gly Asp Phe Leu Glu Ala Val Ala 
    210                 215                 220 

Pro Ile Leu Glu Gln Asn Pro Lys Ala Ile Ala Phe Ile Ala Gly Ser 
225                 230                 235                 240 

Ala Phe Glu Gly Glu Glu Trp Arg Val Val Glu Leu Glu Lys Lys Ile 
                245                 250                 255 

Ser Gln Leu Lys Val Ser Ser Gln Val Arg Arg Met Asp Tyr Tyr Ala 
            260                 265                 270 

Asn Thr Thr Glu Leu Tyr Asn Met Phe Asp Ile Phe Val Leu Pro Ser 
        275                 280                 285 

Thr Asn Pro Asp Pro Leu Pro Thr Val Val Leu Lys Ala Met Ala Cys 
    290                 295                 300 

Gly Lys Pro Val Val Gly Tyr Arg His Gly Gly Val Cys Glu Met Val 
305                 310                 315                 320 

Lys Glu Gly Val Asn Gly Phe Leu Val Thr Pro Asn Ser Pro Leu Asn 
                325                 330                 335 

Leu Ser Lys Val Ile Leu Gln Leu Ser Glu Asn Ile Asn Leu Arg Lys 
            340                 345                 350 

Lys Ile Gly Asn Asn Ser Ile Glu Arg Gln Lys Glu His Phe Ser Leu 
        355                 360                 365 

Lys Ser Tyr Val Lys Asn Phe Ser Lys Val Tyr Thr Ser Leu Lys Val 
    370                 375                 380 

Tyr 
385 

 
           
             19  
             456  
             PRT  
             Streptococcus suis  
             
               misc_feature  
               cps2h  
             
           
            19 

Met Lys Ile Ile Ser Phe Thr Met Val Asn Asn Glu Ser Glu Ile Ile 
1               5                   10                  15 

Glu Ser Phe Ile Arg Tyr Asn Tyr Asn Phe Ile Asp Glu Met Val Ile 
            20                  25                  30 

Ile Asp Asn Gly Cys Thr Asp Asn Thr Met Gln Ile Ile Phe Asn Leu 
        35                  40                  45 

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

Tyr Asn Gln Tyr Arg Leu Asp Asn Lys Tyr Leu Thr Lys Ile Ile Ala 
65                  70                  75                  80 

Glu Lys Asn Pro Asp Leu Ile Ile Pro Leu Asp Ala Asp Glu Phe Leu 
                85                  90                  95 

Thr Ala Asp Ser Asn Pro Arg Lys Leu Leu Glu Gln Leu Asp Leu Glu 
            100                 105                 110 

Lys Ile His Tyr Val Asn Trp Gln Trp Phe Val Met Thr Lys Lys Asp 
        115                 120                 125 

Asp Ile Asn Asp Ser Phe Ile Pro Arg Arg Met Gln Tyr Cys Phe Glu 
    130                 135                 140 

Lys Pro Val Trp His His Ser Asp Gly Lys Pro Val Thr Lys Cys Ile 
145                 150                 155                 160 

Ile Ser Ala Lys Tyr Tyr Lys Lys Met Asn Leu Lys Leu Ser Met Gly 
                165                 170                 175 

His His Thr Val Phe Gly Asn Pro Asn Val Arg Ile Glu His His Asn 
            180                 185                 190 

Asp Leu Lys Phe Ala His Tyr Arg Ala Ile Ser Gln Glu Gln Leu Ile 
        195                 200                 205 

Tyr Lys Thr Ile Cys Tyr Thr Ile Arg Asp Ile Ala Thr Met Glu Asn 
    210                 215                 220 

Asn Ile Glu Thr Ala Gln Arg Thr Asn Gln Met Ala Leu Ile Glu Ser 
225                 230                 235                 240 

Gly Val Asp Met Trp Glu Thr Ala Arg Glu Ala Ser Tyr Ser Gly Tyr 
                245                 250                 255 

Asp Cys Asn Val Ile His Ala Pro Ile Asp Leu Ser Phe Cys Lys Glu 
            260                 265                 270 

Asn Ile Val Ile Lys Tyr Asn Glu Leu Ser Arg Glu Thr Val Ala Glu 
        275                 280                 285 

Arg Val Met Lys Thr Gly Arg Glu Met Ala Val Arg Ala Tyr Asn Val 
    290                 295                 300 

Glu Arg Lys Gln Lys Glu Lys Lys Phe Leu Lys Pro Ile Ile Phe Val 
305                 310                 315                 320 

Leu Asp Gly Leu Lys Gly Asp Glu Tyr Ile His Pro Asn Pro Ser Asn 
                325                 330                 335 

His Leu Thr Ile Leu Thr Glu Met Tyr Asn Val Arg Gly Leu Leu Thr 
            340                 345                 350 

Asp Asn His Gln Ile Lys Phe Leu Lys Val Asn Tyr Arg Leu Ile Ile 
        355                 360                 365 

Thr Pro Asp Phe Ala Lys Phe Leu Pro His Glu Phe Ile Val Val Pro 
    370                 375                 380 

Asp Thr Leu Asp Ile Glu Gln Val Lys Ser Gln Tyr Val Gly Thr Gly 
385                 390                 395                 400 

Val Asp Leu Ser Lys Ile Ile Ser Leu Lys Glu Tyr Arg Lys Glu Ile 
                405                 410                 415 

Gly Phe Ile Gly Asn Leu Tyr Ala Leu Leu Gly Phe Val Pro Asn Met 
            420                 425                 430 

Leu Asn Arg Ile Tyr Leu Tyr Ile Gln Arg Asn Gly Ile Ala Asn Thr 
        435                 440                 445 

Ile Ile Lys Ile Lys Ser Arg Leu 
    450                 455 

 
           
             20  
             410  
             PRT  
             Streptococcus suis  
             
               misc_feature  
               CPS2I  
             
           
            20 

Met Gln Ala Asp Arg Arg Lys Thr Phe Gly Lys Met Arg Ile Arg Ile 
1               5                   10                  15 

Asn Asn Leu Phe Phe Val Ala Ile Ala Phe Met Gly Ile Ile Ile Ser 
            20                  25                  30 

Asn Ser Gln Val Val Leu Ala Ile Gly Lys Ala Ser Val Ile Gln Tyr 
        35                  40                  45 

Leu Ser Tyr Leu Val Leu Ile Leu Cys Ile Val Asn Asp Leu Leu Lys 
    50                  55                  60 

Asn Asn Lys His Ile Val Val Tyr Lys Leu Gly Tyr Leu Phe Leu Ile 
65                  70                  75                  80 

Ile Phe Leu Phe Thr Ile Gly Ile Cys Gln Gln Ile Leu Pro Ile Thr 
                85                  90                  95 

Thr Lys Ile Tyr Leu Ser Ile Ser Met Met Ile Ile Ser Val Leu Ala 
            100                 105                 110 

Thr Leu Pro Ile Ser Leu Ile Lys Asp Ile Asp Asp Phe Arg Arg Ile 
        115                 120                 125 

Ser Asn His Leu Leu Phe Ala Leu Phe Ile Thr Ser Ile Leu Gly Ile 
    130                 135                 140 

Lys Met Gly Ala Thr Met Phe Thr Gly Ala Val Glu Gly Ile Gly Phe 
145                 150                 155                 160 

Ser Gln Gly Phe Asn Gly Gly Leu Thr His Lys Asn Phe Phe Gly Ile 
                165                 170                 175 

Thr Ile Leu Met Gly Phe Val Leu Thr Tyr Leu Ala Tyr Lys Tyr Gly 
            180                 185                 190 

Ser Tyr Lys Arg Thr Asp Arg Phe Ile Leu Gly Leu Glu Leu Phe Leu 
        195                 200                 205 

Ile Leu Ile Ser Asn Thr Arg Ser Val Tyr Leu Ile Leu Leu Leu Phe 
    210                 215                 220 

Leu Phe Leu Val Asn Leu Asp Lys Ile Lys Ile Glu Gln Arg Gln Trp 
225                 230                 235                 240 

Ser Thr Leu Lys Tyr Ile Ser Met Leu Phe Cys Ala Ile Phe Leu Tyr 
                245                 250                 255 

Tyr Phe Phe Gly Phe Leu Ile Thr His Ser Asp Ser Tyr Ala His Arg 
            260                 265                 270 

Val Asn Gly Leu Ile Asn Phe Phe Glu Tyr Tyr Arg Asn Asp Trp Phe 
        275                 280                 285 

His Leu Met Phe Gly Ala Ala Asp Leu Ala Tyr Gly Asp Leu Thr Leu 
    290                 295                 300 

Asp Tyr Ala Ile Arg Val Arg Arg Val Leu Gly Trp Asn Gly Thr Leu 
305                 310                 315                 320 

Glu Met Pro Leu Leu Ser Ile Met Leu Lys Asn Gly Phe Ile Gly Leu 
                325                 330                 335 

Val Gly Tyr Gly Ile Val Leu Tyr Lys Leu Tyr Arg Asn Val Arg Ile 
            340                 345                 350 

Leu Lys Thr Asp Asn Ile Lys Thr Ile Gly Lys Ser Val Phe Ile Ile 
        355                 360                 365 

Val Val Leu Ser Ala Thr Val Glu Asn Tyr Ile Val Asn Leu Ser Phe 
    370                 375                 380 

Val Phe Met Pro Ile Cys Phe Cys Leu Leu Asn Ser Ile Ser Thr Met 
385                 390                 395                 400 

Glu Ser Thr Ile Asn Lys Gln Leu Gln Thr 
                405                 410 

 
           
             21  
             332  
             PRT  
             Streptococcus suis  
             
               misc_feature  
               CPS2J  
             
           
            21 

Met Glu Lys Val Ser Ile Ile Val Pro Ile Phe Asn Thr Glu Lys Tyr 
1               5                   10                  15 

Leu Arg Glu Cys Leu Asp Ser Ile Ile Ser Gln Ser Tyr Thr Asn Leu 
            20                  25                  30 

Glu Ile Leu Leu Ile Asp Asp Gly Ser Ser Asp Ser Ser Thr Asp Ile 
        35                  40                  45 

Cys Leu Glu Tyr Ala Glu Gln Asp Gly Arg Ile Lys Leu Phe Arg Leu 
    50                  55                  60 

Pro Asn Gly Gly Val Ser Asn Ala Arg Asn Tyr Gly Ile Lys Asn Ser 
65                  70                  75                  80 

Thr Ala Asn Tyr Ile Met Phe Val Asp Ser Asp Asp Ile Val Asp Gly 
                85                  90                  95 

Asn Ile Val Glu Ser Leu Tyr Thr Cys Leu Lys Glu Asn Asp Ser Asp 
            100                 105                 110 

Leu Ser Gly Gly Leu Leu Ala Thr Phe Asp Gly Asn Tyr Gln Glu Ser 
        115                 120                 125 

Glu Leu Gln Lys Cys Gln Ile Asp Leu Glu Glu Ile Lys Glu Val Arg 
    130                 135                 140 

Asp Leu Gly Asn Glu Asn Phe Pro Asn His Tyr Met Ser Gly Ile Phe 
145                 150                 155                 160 

Asn Ser Pro Cys Cys Lys Leu Tyr Lys Asn Ile Tyr Ile Asn Gln Gly 
                165                 170                 175 

Phe Asp Thr Glu Gln Trp Leu Gly Glu Asp Leu Leu Phe Asn Leu Asn 
            180                 185                 190 

Tyr Leu Lys Asn Ile Lys Lys Val Arg Tyr Val Asn Arg Asn Leu Tyr 
        195                 200                 205 

Phe Ala Arg Arg Ser Leu Gln Ser Thr Thr Asn Thr Phe Lys Tyr Asp 
    210                 215                 220 

Val Phe Ile Gln Leu Glu Asn Leu Glu Glu Lys Thr Phe Asp Leu Phe 
225                 230                 235                 240 

Val Lys Ile Phe Gly Gly Gln Tyr Glu Phe Ser Val Phe Lys Glu Thr 
                245                 250                 255 

Leu Gln Trp His Ile Ile Tyr Tyr Ser Leu Leu Met Phe Lys Asn Gly 
            260                 265                 270 

Asp Glu Ser Leu Pro Lys Lys Leu His Ile Phe Lys Tyr Leu Tyr Asn 
        275                 280                 285 

Arg His Ser Leu Asp Thr Leu Ser Ile Lys Arg Thr Ser Ser Val Phe 
    290                 295                 300 

Lys Arg Ile Cys Lys Leu Ile Val Ala Asn Asn Leu Phe Lys Ile Phe 
305                 310                 315                 320 

Leu Asn Thr Leu Ile Arg Glu Glu Lys Asn Asn Asp 
                325                 330 

 
           
             22  
             332  
             PRT  
             Streptococcus suis  
             
               misc_feature  
               CPS2K  
             
           
            22 

Met Ile Asn Ile Ser Ile Ile Val Pro Ile Tyr Asn Val Glu Gln Tyr 
1               5                   10                  15 

Leu Ser Lys Cys Ile Asn Ser Ile Val Asn Gln Thr Tyr Lys His Ile 
            20                  25                  30 

Glu Ile Leu Leu Val Asn Asp Gly Ser Thr Asp Asn Ser Glu Glu Ile 
        35                  40                  45 

Cys Leu Ala Tyr Ala Lys Lys Asp Ser Arg Ile Arg Tyr Phe Lys Lys 
    50                  55                  60 

Glu Asn Gly Gly Leu Ser Asp Ala Arg Asn Tyr Gly Ile Ser Arg Ala 
65                  70                  75                  80 

Lys Gly Asp Tyr Leu Ala Phe Ile Asp Ser Asp Asp Phe Ile His Ser 
                85                  90                  95 

Glu Phe Ile Gln Arg Leu His Glu Ala Ile Glu Arg Glu Asn Ala Leu 
            100                 105                 110 

Val Ala Val Ala Gly Tyr Asp Arg Val Asp Ala Ser Gly His Phe Leu 
        115                 120                 125 

Thr Ala Glu Pro Leu Pro Thr Asn Gln Ala Val Leu Ser Gly Arg Asn 
    130                 135                 140 

Val Cys Lys Lys Leu Leu Glu Ala Asp Gly His Arg Phe Val Val Ala 
145                 150                 155                 160 

Trp Asn Lys Leu Tyr Lys Lys Glu Leu Phe Asp Phe Arg Phe Glu Lys 
                165                 170                 175 

Gly Lys Ile His Glu Asp Glu Tyr Phe Thr Tyr Arg Leu Leu Tyr Glu 
            180                 185                 190 

Leu Glu Lys Val Ala Ile Val Lys Glu Cys Leu Tyr Tyr Tyr Val Asp 
        195                 200                 205 

Arg Glu Asn Ser Ile Ile Thr Ser Ser Met Thr Asp His Arg Phe His 
    210                 215                 220 

Cys Leu Leu Glu Phe Gln Asn Glu Arg Met Asp Phe Tyr Glu Ser Arg 
225                 230                 235                 240 

Gly Asp Lys Glu Leu Leu Leu Glu Cys Tyr Arg Ser Phe Leu Ala Phe 
                245                 250                 255 

Ala Val Leu Phe Leu Gly Lys Tyr Asn His Trp Leu Ser Lys Gln Gln 
            260                 265                 270 

Lys Lys Leu Gln Thr Leu Phe Arg Ile Val Tyr Lys Gln Leu Lys Gln 
        275                 280                 285 

Asn Lys Arg Leu Ala Leu Leu Met Asn Ala Tyr Tyr Leu Val Gly Cys 
    290                 295                 300 

Leu His Leu Asn Phe Ser Val Phe Leu Lys Thr Gly Lys Asp Lys Ile 
305                 310                 315                 320 

Gln Glu Arg Leu Arg Arg Ser Glu Ser Ser Thr Arg 
                325                 330 

 
           
             23  
             467  
             PRT  
             Streptococcus suis  
             
               misc_feature  
               CPS2O  
             
           
            23 

Met Ser Lys Lys Ser Ile Val Val Ser Gly Leu Val Tyr Thr Ile Gly 
1               5                   10                  15 

Thr Ile Leu Val Gln Gly Leu Ala Phe Ile Thr Leu Pro Ile Tyr Thr 
            20                  25                  30 

Arg Val Ile Ser Gln Glu Val Tyr Gly Gln Phe Ser Leu Tyr Asn Ser 
        35                  40                  45 

Trp Val Gly Leu Val Gly Leu Phe Ile Gly Leu Gln Leu Gly Gly Ala 
    50                  55                  60 

Phe Gly Pro Gly Trp Val His Phe Arg Glu Lys Phe Asp Asp Phe Val 
65                  70                  75                  80 

Ser Thr Leu Met Val Ser Ser Ile Ala Phe Phe Leu Pro Ile Phe Gly 
                85                  90                  95 

Leu Ser Phe Leu Leu Ser Gln Pro Leu Ser Leu Leu Phe Gly Leu Pro 
            100                 105                 110 

Asp Trp Val Val Pro Leu Ile Phe Leu Gln Ser Leu Met Ile Val Val 
        115                 120                 125 

Gln Gly Phe Phe Thr Thr Tyr Leu Val Gln Arg Gln Gln Ser Met Trp 
    130                 135                 140 

Thr Leu Pro Leu Ser Val Leu Ser Ala Val Ile Asn Thr Ala Leu Ser 
145                 150                 155                 160 

Leu Phe Leu Thr Phe Pro Met Glu Asn Asp Phe Ile Ala Arg Val Met 
                165                 170                 175 

Ala Asn Pro Ala Thr Thr Gly Val Leu Ala Cys Val Ser Xaa Trp Phe 
            180                 185                 190 

Ser Gln Lys Lys Asn Gly Leu His Phe Arg Lys Asp Tyr Leu Arg Tyr 
        195                 200                 205 

Gly Leu Ser Ile Ser Ile Pro Leu Ile Phe His Gly Leu Gly His Asn 
    210                 215                 220 

Val Leu Asn Gln Phe Asp Arg Ile Met Leu Gly Lys Met Leu Thr Leu 
225                 230                 235                 240 

Ser Asp Val Ala Leu Tyr Ser Phe Gly Tyr Thr Leu Ala Ser Ile Leu 
                245                 250                 255 

Gln Ile Val Phe Ser Ser Leu Asn Thr Val Trp Cys Pro Trp Tyr Phe 
            260                 265                 270 

Glu Lys Lys Arg Gly Ala Asp Lys Asp Leu Leu Ser Tyr Val Arg Tyr 
        275                 280                 285 

Tyr Leu Ala Ile Gly Leu Phe Val Thr Phe Gly Phe Leu Thr Ile Tyr 
    290                 295                 300 

Pro Arg Leu Ala Met Leu Leu Gly Gly Ser Glu Tyr Arg Phe Ser Met 
305                 310                 315                 320 

Gly Phe Ile Pro Met Ile Ile Val Gly Val Phe Phe Val Phe Leu Tyr 
                325                 330                 335 

Ser Phe Pro Ala Asn Ile Gln Phe Tyr Ser Gly Asn Thr Lys Phe Leu 
            340                 345                 350 

Pro Ile Gly Thr Phe Ile Ala Gly Val Leu Asn Ile Ser Val His Phe 
        355                 360                 365 

Val Leu Ile Pro Thr Lys Asn Leu Trp Cys Cys Phe Ala Thr Thr Ala 
    370                 375                 380 

Ser Tyr Leu Leu Leu Leu Val Leu His Tyr Phe Val Ala Lys Lys Lys 
385                 390                 395                 400 

Tyr Ala Tyr Asp Glu Val Ala Ile Ser Thr Phe Val Lys Val Ile Ala 
                405                 410                 415 

Leu Val Val Val Tyr Thr Gly Leu Met Thr Val Phe Val Gly Ser Ile 
            420                 425                 430 

Trp Ile Arg Trp Ser Leu Gly Ile Ala Val Leu Val Val Tyr Ala Ile 
        435                 440                 445 

Tyr Phe Arg Lys Glu Leu Thr Val Ala Leu Asn Thr Phe Arg Glu Lys 
    450                 455                 460 

Arg Ser Lys 
465 

 
           
             24  
             338  
             PRT  
             Streptococcus suis  
             
               misc_feature  
               CPS2P  
             
           
            24 

Met Val Tyr Ile Ile Ala Glu Ile Gly Cys Asn His Asn Gly Asp Val 
1               5                   10                  15 

His Leu Ala Arg Lys Met Val Glu Val Ala Val Asp Cys Gly Val Asp 
            20                  25                  30 

Ala Val Lys Phe Gln Thr Glu Lys Ala Asp Leu Leu Ile Ser Lys Tyr 
        35                  40                  45 

Ala Pro Lys Ala Glu Tyr Gln Lys Ile Thr Thr Gly Glu Ser Asp Ser 
    50                  55                  60 

Gln Leu Glu Met Thr Arg Arg Leu Glu Leu Ser Phe Glu Glu Tyr Leu 
65                  70                  75                  80 

Asp Leu Arg Asp Tyr Cys Leu Glu Lys Gly Val Asp Val Phe Ser Thr 
                85                  90                  95 

Pro Glu Asp Glu Glu Ser Leu Asp Phe Leu Ile Ser Thr Asp Met Pro 
            100                 105                 110 

Val Tyr Lys Ile Pro Ser Gly Glu Ile Thr Asn Leu Pro Tyr Leu Glu 
        115                 120                 125 

Lys Ile Gly Arg Gln Ala Lys Lys Val Ile Leu Ser Thr Gly Met Ala 
    130                 135                 140 

Val Met Asp Glu Ile His Gln Ala Val Lys Ile Leu Gln Glu Asn Gly 
145                 150                 155                 160 

Thr Thr Asp Ile Ser Ile Leu His Cys Thr Thr Glu Tyr Pro Thr Pro 
                165                 170                 175 

Tyr Pro Ala Leu Asn Leu Asn Val Leu His Thr Leu Lys Lys Glu Phe 
            180                 185                 190 

Pro Asn Leu Thr Ile Gly Tyr Ser Asp His Ser Val Gly Ser Glu Val 
        195                 200                 205 

Pro Ile Ala Ala Ala Ala Met Gly Ala Glu Leu Ile Glu Lys His Phe 
    210                 215                 220 

Thr Leu Asp Asn Glu Met Glu Gly Pro Asp His Lys Ala Ser Ala Thr 
225                 230                 235                 240 

Pro Asp Ile Leu Ala Ala Leu Val Lys Gly Val Arg Ile Val Glu Gln 
                245                 250                 255 

Ser Leu Gly Lys Phe Glu Lys Glu Pro Glu Glu Val Glu Val Arg Asn 
            260                 265                 270 

Lys Ile Val Ala Glu Lys Ser Ile Val Ala Lys Lys Ala Ile Ala Lys 
        275                 280                 285 

Gly Glu Val Phe Thr Glu Glu Asn Ile Thr Val Lys Arg Pro Gly Asn 
    290                 295                 300 

Gly Ile Ser Pro Met Glu Trp Tyr Lys Val Leu Gly Gln Val Ser Glu 
305                 310                 315                 320 

Gln Asp Phe Glu Glu Asp Gln Asn Ile Cys His Ser Ala Phe Glu Asn 
                325                 330                 335 
Gln Met 

 
           
             25  
             170  
             PRT  
             Streptococcus suis  
             
               misc_feature  
               CPS2Q  
             
           
            25 

Met Lys Lys Ile Cys Phe Val Thr Gly Ser Arg Ala Glu Tyr Gly Ile 
1               5                   10                  15 

Met Arg Arg Leu Leu Ser Tyr Leu Gln Asp Asp Pro Glu Met Glu Leu 
            20                  25                  30 

Asp Leu Val Val Ala Thr Met His Leu Glu Glu Lys Tyr Gly Met Thr 
        35                  40                  45 

Val Lys Asp Ile Glu Ala Asp Lys Arg Arg Ile Val Lys Arg Ile Pro 
    50                  55                  60 

Leu His Leu Thr Asp Thr Ser Lys Gln Thr Ile Val Lys Ser Leu Ala 
65                  70                  75                  80 

Thr Leu Thr Glu Gln Leu Thr Val Leu Phe Glu Glu Val Gln Tyr Asp 
                85                  90                  95 

Leu Val Leu Ile Leu Gly Asp Arg Tyr Glu Met Leu Pro Val Ala Asn 
            100                 105                 110 

Ala Ala Leu Leu Tyr Asn Ile Pro Ile Cys His Ile His Gly Gly Glu 
        115                 120                 125 

Lys Thr Met Gly Asn Phe Asp Glu Ser Ile Arg His Ala Ile Thr Lys 
    130                 135                 140 

Met Ser His Leu His Leu Thr Ser Thr Asp Glu Phe Arg Asn Arg Val 
145                 150                 155                 160 

Ile Gln Leu Gly Glu Asn Pro Thr Met Tyr 
                165                 170 

 
           
             26  
             184  
             PRT  
             Streptococcus suis  
             
               misc_feature  
               CPS2R  
             
           
            26 

Met Glu Leu Gly Ile Asp Phe Ala Glu Asp Tyr Tyr Val Val Leu Phe 
1               5                   10                  15 

His Pro Val Thr Leu Glu Asp Asn Thr Ala Glu Glu Gln Thr Gln Ala 
            20                  25                  30 

Leu Leu Asp Ala Leu Lys Glu Asp Gly Ser Gln Cys Leu Ile Ile Gly 
        35                  40                  45 

Ser Asn Ser Asp Thr His Ala Asp Lys Ile Met Glu Leu Met His Glu 
    50                  55                  60 

Phe Val Lys Gln Asp Ser Asp Ser Tyr Ile Phe Thr Ser Leu Pro Thr 
65                  70                  75                  80 

Arg Tyr Tyr His Ser Leu Val Lys His Ser Gln Gly Leu Ile Gly Asn 
                85                  90                  95 

Ser Ser Ser Gly Leu Ile Glu Val Pro Ser Leu Gln Val Pro Thr Leu 
            100                 105                 110 

Asn Ile Gly Asn Arg Gln Phe Gly Arg Leu Ser Gly Pro Ser Val Val 
        115                 120                 125 

His Val Gly Thr Ser Lys Glu Ala Ile Val Gly Gly Leu Gly Gln Leu 
    130                 135                 140 

Arg Asp Val Ile Asp Phe Thr Asn Pro Phe Glu Gln Pro Asp Ser Ala 
145                 150                 155                 160 

Leu Gln Gly Tyr Arg Ala Ile Lys Glu Phe Leu Ser Val Gln Ala Ser 
                165                 170                 175 

Thr Met Lys Glu Phe Tyr Asp Arg 
            180 

 
           
             27  
             208  
             PRT  
             Streptococcus suis  
             
               misc_feature  
               CPS2S  
             
           
            27 

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

Leu Pro Trp Leu Asp Arg Thr Arg Tyr Glu Leu Ile Gly Tyr Phe Glu 
            20                  25                  30 

Asp Lys Pro Ile Ser Asp Tyr Arg Gly Tyr Pro Val Phe Gly Pro Leu 
        35                  40                  45 

Gln Asp Val Leu Thr Tyr Leu Asp Asp Gly Lys Val Asp Ala Val Phe 
    50                  55                  60 

Val Thr Ile Gly Asp Asn Val Lys Arg Lys Glu Ile Phe Asp Leu Leu 
65                  70                  75                  80 

Ala Lys Asp His Tyr Asp Ala Leu Phe Asn Ile Ile Ser Glu Gln Ala 
                85                  90                  95 

Asn Ile Phe Ser Pro Asp Ser Ile Lys Gly Arg Gly Val Phe Ile Gly 
            100                 105                 110 

Phe Ser Ser Phe Val Gly Ala Asp Ser Tyr Val Tyr Asp Asn Cys Ile 
        115                 120                 125 

Ile Asn Thr Gly Ala Ile Val Glu His His Thr Thr Val Glu Ala His 
    130                 135                 140 

Cys Asn Ile Thr Pro Gly Val Thr Ile Asn Gly Leu Cys Arg Ile Gly 
145                 150                 155                 160 

Glu Ser Thr Tyr Ile Gly Ser Gly Ser Thr Val Ile Gln Cys Ile Glu 
                165                 170                 175 

Ile Ala Pro Tyr Thr Thr Leu Gly Ala Gly Thr Val Val Leu Lys Ser 
            180                 185                 190 

Leu Thr Glu Ser Gly Thr Tyr Val Gly Val Pro Ala Arg Lys Ile Lys 
        195                 200                 205 

 
           
             28  
             410  
             PRT  
             Streptococcus suis  
             
               misc_feature  
               CPS2T  
             
           
            28 

Met Glu Pro Ile Cys Leu Ile Pro Ala Arg Ser Gly Ser Lys Gly Leu 
1               5                   10                  15 

Pro Asn Lys Asn Met Leu Phe Leu Asp Gly Val Pro Met Ile Phe His 
            20                  25                  30 

Thr Ile Arg Ala Ala Ile Glu Ser Gly Cys Phe Lys Lys Glu Asn Ile 
        35                  40                  45 

Tyr Val Ser Thr Asp Ser Glu Val Tyr Lys Glu Ile Cys Glu Thr Thr 
    50                  55                  60 

Gly Val Gln Val Leu Met Arg Pro Ala Asp Leu Ala Thr Asp Phe Thr 
65                  70                  75                  80 

Thr Ser Phe Gln Leu Asn Glu His Phe Leu Gln Asp Phe Ser Asp Asp 
                85                  90                  95 

Gln Val Phe Val Leu Leu Gln Val Thr Ser Pro Leu Arg Ser Gly Lys 
            100                 105                 110 

His Val Lys Glu Ala Met Glu Leu Tyr Gly Lys Gly Gln Ala Asp His 
        115                 120                 125 

Val Val Ser Phe Thr Lys Val Asp Lys Ser Pro Thr Leu Phe Ser Thr 
    130                 135                 140 

Leu Asp Glu Asn Gly Phe Ala Lys Asp Ile Ala Gly Leu Gly Gly Ser 
145                 150                 155                 160 

Tyr Arg Arg Gln Asp Glu Lys Thr Leu Tyr Tyr Pro Asn Gly Ala Ile 
                165                 170                 175 

Tyr Ile Ser Ser Lys Gln Ala Tyr Leu Ala Asp Lys Thr Tyr Phe Ser 
            180                 185                 190 

Glu Lys Thr Ala Ala Tyr Val Met Thr Lys Glu Asp Ser Ile Asp Val 
        195                 200                 205 

Asp Asp His Phe Asp Phe Thr Gly Val Ile Gly Arg Ile Tyr Phe Asp 
    210                 215                 220 

Tyr Gln Arg Arg Glu Gln Gln Asn Lys Pro Phe Tyr Lys Arg Glu Leu 
225                 230                 235                 240 

Lys Arg Leu Cys Glu Gln Arg Val His Asp Ser Leu Val Ile Gly Asp 
                245                 250                 255 

Ser Arg Leu Leu Ala Leu Leu Leu Asp Gly Phe Asp Asn Ile Ser Ile 
            260                 265                 270 

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

Ala Thr Pro Ile Lys Lys Val Leu Leu Ser Leu Gly Val Asn Asp Leu 
    290                 295                 300 

Ile Thr Asp Tyr Pro Leu His Met Ile Glu Asp Thr Ile Arg Gln Leu 
305                 310                 315                 320 

Met Glu Ser Leu Val Ser Lys Ala Glu Gln Val Glu Val Thr Thr Ile 
                325                 330                 335 

Ala Tyr Thr Leu Phe Arg Asp Ser Val Ser Asn Glu Glu Thr Val Gln 
            340                 345                 350 

Leu Asn Asp Val Ile Val Gln Ser Ala Ser Glu Leu Gly Ile Ser Val 
        355                 360                 365 

Ile Asp Leu Asn Glu Val Val Glu Lys Glu Ala Met Leu Asp Tyr Gln 
    370                 375                 380 

Tyr Thr Asn Asp Gly Leu His Phe Asn Gln Ile Gly Gln Glu Arg Val 
385                 390                 395                 400 

Asn Gln Leu Ile Leu Thr Ser Leu Thr Arg 
                405                 410 

 
           
             29  
             6992  
             DNA  
             Streptococcus suis  
             
               misc_feature  
               CPS1  
             
           
            29 

atcgccaaac gaaattggca ttatttgata tgatagcagt tgcaatttct gcaatcttaa     60 

caagtcatat accaaatgct gatttaaatc gttctggaat ttttatcata atgatggttc    120 

attattttgc attttttata tctcgtatgc cagttgaatt tgagtataga ggtaatctga    180 

tagagtttga aaaaacattt aactatagta taatatttgc aatttttctt acggcagtat    240 

catttttgtt ggagaataat ttcgcacttt caagacgtgg tgccgtgtat ttcacattaa    300 

taaacttcgt tttggtatac ctatttaacg taattattaa gcagtttaag gatagctttc    360 

tattttcgac aatctatcaa aaaaagacga ttctaattac aacggctgaa cgatgggaaa    420 

atatgcaagt tttatttgaa tcacataaac aaattcaaaa aaatcttgtt gcattggtag    480 

ttttaggtac agaaatagat aaaattaatt tatcattacc gctctattat tctgtggaag    540 

aagctataga gttttcaaca agggaagtgg tcgaccacgt ctttataaat ctaccaagtg    600 

agtttttaga cgtaaagcaa ttcgtttcag attttgagtt gttaggtatt gatgtaagcg    660 

ttgatattaa ttcattcggt tttactgcgt tgaaaaacaa aaaaatccaa ctgctaggtg    720 

accatagcat tgtaactttt tccacaaatt tttataagcc tagtcatatc atgatgaaac    780 

gacttttgga tatactcgga gcggtagtcg ggttaattat ttgtggtata gtttctattt    840 

tgttagttcc aattattcgt agagatggtg gaccggctat ttttgctcag aaacgagttg    900 

gacagaatgg acgcatattt acattctaca agtttcgatc gatgtatgtt gatgctgagg    960 

agcgcaaaaa agacttgctc agccaaaacc agatgcaagg gtgggtatgt tttaaaatgg   1020 

gaaaaacgat cctagaatta ctccaattgg acatttcata cgcaaaaaca agtttagacg   1080 

agttaccaca gttttataat gttttaattg gcgatatgag tctagttggt acacgtccac   1140 

ctacagttga tgaatttgaa aaatatactc ctggtcaaaa gagacgattg agttttaaac   1200 

cagggattac aggtctctgg caggttagtg gtcgtagtaa tatcacagac ttcgacgacg   1260 

tagttcggtt ggacttagca tacattgata attggactat ctggtcagat attaaaattt   1320 

tattaaagac agtgaaagtt gtattgttga gagagggaag taagtaaaag tatatgaaag   1380 

tttgtttggt cggttcttca gggggacatt tgactcactt gtatttgtta aaaccgtttt   1440 

ggaaggaaga agaacgtttt tgggtaacat ttgataaaga ggatgcaaga agtcttttga   1500 

agaatgaaaa aatgtatcca tgttactttc caacaaatcg caatctcatt aatttagtga   1560 

aaaatacttt cttagctttc aaaattttac gtgatgagaa accagatgtt attatttcat   1620 

ctggtgcggc cgttgctgtc cccttctttt acatcggaaa actatttgga gcaaagacga   1680 

tttatattga agtatttgat cgagttaata aatctacatt aactggaaaa ctagtttatc   1740 

ccgtaacaga tatttttatt gttcagtggg aagaaatgaa gaaggtatat cctaaatcta   1800 

ttaacttggg gagtattttt taatgatttt tgtaacagta ggaactcatg aacaacagtt   1860 

taatcgattg ataaaagaga ttgatttatt gaaaaaaaat ggaagtataa ccgacgaaat   1920 

atttattcaa acaggatatt ctgactatat tccagaatat tgcaagtata aaaaatttct   1980 

cagttacaaa gaaatggaac aatatattaa caaatcagaa gtagttattt gccacggagg   2040 

ccccgctact tttatgaatt cattatccaa aggaaaaaaa caattattgt ttcctagaca   2100 

aaaaaagtat ggtgaacatg taaatgatca tcaagtagag tttgtaagaa gaattttaca   2160 

agataataat attttattta tagaaaatat agatgatttg tttgaaaaaa ttattgaagt   2220 

ttctaagcaa actaacttta catcaaataa taattttttt tgtgaaagat taaaacaaat   2280 

agttgaaaaa tttaatgagg atcaagaaaa tgaataataa aaaagatgca tatttgataa   2340 

tggcttatca taatttttct cagattttac tggagaggga tacagatatt atcatcttct   2400 

ctcaggagaa tgcacaccat tagttccttc agaatacctg tataattatt ttaaatattc   2460 

tcaggattta tatgttgaat ttacaaaaga tgagcaaaaa tataaagaaa ataggatata   2520 

tgaacgagtt aaatgttaca gattatttcc taatatatca gaaaaaacta ttgataatgt   2580 

actgtttaga attttattaa gaatgtatcg agcttttgaa tactatttac aaagattgtt   2640 

gtttattgat agaataaaaa acatggtcta agaataagat ttggttctaa ttgggtttcg   2700 

cttccacatg attttgtggc aattctttta tcaaatgaaa acgaaacagc ttatttattt   2760 

aagtaatcta aatgtccaga tgaactattt atacagacaa ttatagaaaa atatgaattt   2820 

tcaaatagat tatctaaata tggaaattta agatatataa agtggaaaaa atcaacatct   2880 

tctcctattg tctttacaga tgattctatt gatgaattgc taaatgcaag aaatttaggt   2940 

tttttatttg ctagaaagtt aaaaatagaa aataaatcta aatttaaaga aattattact   3000 

aaaaaataaa atagttgatt ttgtgagagt aatgtatgtt taaattattt aaatatgacc   3060 

cggaatattt tatttttaag tacttctggt tgattatttt tattccagag caaaagtatg   3120 

tatttttatt aatttttatg aatttaattt tatttcatat aaaatttttg aaaactaagc   3180 

taatattaaa aaatgaaatt ttattgtttt tattatggtc tatattatgt tttgtttcag   3240 

tagtcacaag tatgtttgtt gaaataaatt ttgaaagatt atttgcagat tttactgctc   3300 

ccataatttg gattattgca ataatgtatt ataatttgta ttcatttata aatattgatt   3360 

ataaaaaatt aaaaaatagt atctttttta gttttttagt tttattaggt atatctgcat   3420 

tgtatattat tcaaaatggg aaagatattg tatttttaga cagacacctt ataggactag   3480 

actatcttat aacaggcgtc aaaacaaggt tggttggctt tatgaactat cctacgttaa   3540 

ataccactac aattatagtt tcaattccgt taatctttgc acttataaaa aataaaatgc   3600 

aacaattttt tttcttgtgt cttgctttta taccgatcta tttaagtgga tcgagaattg   3660 

gtagtttatc gctagcaata ttaattatat gcttgttatg gagatatata ggtggaaaat   3720 

ttgcttggat aaaaaagcta atagtaatat ttgtaatact acttattatt ttaaatactg   3780 

aattgcttta ccatgaaatt ttggctgttt ataattctag agaatcaagt aacgaagcta   3840 

gatttattat ttatcaagga agtattgata aagtattaga aaacaatatt ttatttggat   3900 

atggaatatc cgaatattca gttacgggaa cttggctcgg aagtcattca ggctatatat   3960 

cattttttta taaatcagga atagttgggt tgattttact gatgttttct tttttttatg   4020 

ttataaaaaa aagttatgga gttaatgggg aaacagcact attttatttt acatcattag   4080 

ccatattttt catatatgaa acaatagatc cgattattat tatattagta ctattctttt   4140 

cttcaatagg tatttggaat aatataaatt ttaaaaagga tatggagaca aaaaatgaat   4200 

gatttaattt cagttattgt accaatttat aatgtccaag attatcttga taaatgtatt   4260 

aacagtatta ttaaccaaac atatactaat ttagaggtta ttctcgtaaa tgatggaagt   4320 

actgatgatt ctgagaaaat ttgcttaaac tatatgaaga acgatggaag aattaaatat   4380 

tacaagaaaa ttaatggcgg tctagcagat gctcgaaatt tcggactaga acatgcaaca   4440 

ggtaaatata ttgcttttgt cgattctgat gactatatag aagttgcaat gttcgagaga   4500 

atgcatgata atataactga gtataatgcc gatatagcag agatagattt ttgtttagta   4560 

gacgaaaacg ggtatacaaa gaaaaaaaga aatagtaatt ttcatgtctt aacgagagaa   4620 

gagactgtaa aagaattttt gtcaggatct aatatagaaa ataatgtttg gtgcaagctt   4680 

tattcacgag atattataaa agatataaaa ttccaaatta ataatagaag tattggtgag   4740 

gatttgcttt ttaatttgga ggtcttgaac aatgtaacac gtgtagtagt tgatactaga   4800 

gaatattatt ataattatgt cattcgtaac agttcgctta ttaatcagaa attctctata   4860 

aataatattg atttagtcac aagattggag aattacccct ttaagttaaa aagagagttt   4920 

agtcattatt ttgatgcaaa agttattaaa gagaaggtta aatgtttaaa caaaatgtat   4980 

tcaacagatt gtttggataa tgagttcttg ccaatattag agtcttatcg aaaagaaata   5040 

cgtagatatc catttattaa agcgaaaaga tatttatcaa gaaagcattt agttacgttg   5100 

tatttgatga aattttcgcc taaactatat gtaatgttat ataagaaatt tcaaaagcag   5160 

tagaggtaaa aatggataaa attagtgtta ttgttccagt ttataatgta gataaatatt   5220 

taagtagttg tatagaaagc attattaatc aaaattataa aaatatagaa atattattga   5280 

tagatgatgg ctctgtagat gattctgcta aaatatgcaa ggaatatgca gaaaaagata   5340 

aaagagtaaa aatttttttc actaatcata gtggagtatc aaatgctaga aatcatggaa   5400 

taaagcggag tacagctgaa tatattatgt ttgttgactc tgatgatgtt gttgatagta   5460 

gattagtaga aaaattatat tttaatatta taaaaagtag aagtgattta tctggttgtt   5520 

tgtacgctac tttttcagaa aatataaata attttgaagt gaataatcca aatattgatt   5580 

ttgaagcaat taataccgtg caggacatgg gagaaaaaaa ttttatgaat ttgtatataa   5640 

ataatatttt ttctactcct gtttgtaaac tatataagaa aagatacata acagatcttt   5700 

ttcaagagaa tcaatggtta ggagaagatt tactttttaa tctgcattat ttaaagaata   5760 

tagatagagt tagttatttg actgaacatc tttattttta taggagaggt atactaagta   5820 

cagtaaattc ttttaaagaa ggtgtgtttt tgcaattgga aaatttgcaa aaacaagtga   5880 

tagtattgtt taagcaaata tatggtgagg attttgacgt atcaattgtt aaagatacta   5940 

tacgttggca agtattttat tatagcttac taatgtttaa atacggaaaa cagtctattt   6000 

ttgacaaatt tttaattttt agaaatcttt ataaaaaata ttattttaac ttgttaaaag   6060 

tatctaacaa aaattctttg tctaaaaatt tttgtataag aattgtttcg aacaaagttt   6120 

ttaaaaaaat attatggtta taataggaag atatcatgga tactattagt aaaatttcta   6180 

taattgtacc tatatataat gtagaaaaat atttatctaa atgtatagat agcattgtaa   6240 

atcagaccta caaacatata gagattcttc tggtgaatga cggtagtacg gataattcgg   6300 

aagaaatttg tttagcatat gcgaagaaag atagtcgcat tcgttatttt aaaaaagaga   6360 

acggcgggct atcagatgcc cgtaattatg gcataagtcg cgccaagggt gactacttag   6420 

cttttataga ctcagatgat tttattcatt cggagttcat ccaacgttta cacgaagcaa   6480 

ttgagagaga gaatgccctt gtggcagttg ctggttatga tagggtagat gcttcggggc   6540 

atttcttaac agcagagccg cttcctacaa atcaggctgt tctgagcggc aggaatgttt   6600 

gtaaaaagct gctagaggcg gatggtcatc gctttgtggt ggcctgtaat aaactctata   6660 

aaaaagaact atttgaagat tttcgatttg aaaagggtaa gattcatgaa gatgaatact   6720 

tcacttatcg cttgctctat gagttagaaa aagttgcaat agttaaggag tgcttgtact   6780 

attatgttga ccgagaaaat agtatcacaa cttctagcat gactgaccat cgcttccatt   6840 

gcctactgga atttcaaaat gaacgaatgg acttctatga aagtagagga gataaagagc   6900 

tcttactaga gtgttatcgt tcatttttag cctttgctgt tttgttttta ggcaaatata   6960 

atcattggtt gagcaaacag caaaagaagc tt                                 6992 

 
           
             30  
             454  
             PRT  
             Streptococcus suis  
             
               misc_feature  
               CPS1E  
             
           
            30 

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

Ala Ile Leu Thr Ser His Ile Pro Asn Ala Asp Leu Asn Arg Ser Gly 
            20                  25                  30 

Ile Phe Ile Ile Met Met Val His Tyr Phe Ala Phe Phe Ile Ser Arg 
        35                  40                  45 

Met Pro Val Glu Phe Glu Tyr Arg Gly Asn Leu Ile Glu Phe Glu Lys 
    50                  55                  60 

Thr Phe Asn Tyr Ser Ile Ile Phe Ala Ile Phe Leu Thr Ala Val Ser 
65                  70                  75                  80 

Phe Leu Leu Glu Asn Asn Phe Ala Leu Ser Arg Arg Gly Ala Val Tyr 
                85                  90                  95 

Phe Thr Leu Ile Asn Phe Val Leu Val Tyr Leu Phe Asn Val Ile Ile 
            100                 105                 110 

Lys Gln Phe Lys Asp Ser Phe Leu Phe Ser Thr Ile Tyr Gln Lys Lys 
        115                 120                 125 

Thr Ile Leu Ile Thr Thr Ala Glu Arg Trp Glu Asn Met Gln Val Leu 
    130                 135                 140 

Phe Glu Ser His Lys Gln Ile Gln Lys Asn Leu Val Ala Leu Val Val 
145                 150                 155                 160 

Leu Gly Thr Glu Ile Asp Lys Ile Asn Leu Ser Leu Pro Leu Tyr Tyr 
                165                 170                 175 

Ser Val Glu Glu Ala Ile Glu Phe Ser Thr Arg Glu Val Val Asp His 
            180                 185                 190 

Val Phe Ile Asn Leu Pro Ser Glu Phe Leu Asp Val Lys Gln Phe Val 
        195                 200                 205 

Ser Asp Phe Glu Leu Leu Gly Ile Asp Val Ser Val Asp Ile Asn Ser 
    210                 215                 220 

Phe Gly Phe Thr Ala Leu Lys Asn Lys Lys Ile Gln Leu Leu Gly Asp 
225                 230                 235                 240 

His Ser Ile Val Thr Phe Ser Thr Asn Phe Tyr Lys Pro Ser His Ile 
                245                 250                 255 

Met Met Lys Arg Leu Leu Asp Ile Leu Gly Ala Val Val Gly Leu Ile 
            260                 265                 270 

Ile Cys Gly Ile Val Ser Ile Leu Leu Val Pro Ile Ile Arg Arg Asp 
        275                 280                 285 

Gly Gly Pro Ala Ile Phe Ala Gln Lys Arg Val Gly Gln Asn Gly Arg 
    290                 295                 300 

Ile Phe Thr Phe Tyr Lys Phe Arg Ser Met Tyr Val Asp Ala Glu Glu 
305                 310                 315                 320 

Arg Lys Lys Asp Leu Leu Ser Gln Asn Gln Met Gln Gly Trp Val Cys 
                325                 330                 335 

Phe Lys Met Gly Lys Thr Ile Leu Glu Leu Leu Gln Leu Asp Ile Ser 
            340                 345                 350 

Tyr Ala Lys Thr Ser Leu Asp Glu Leu Pro Gln Phe Tyr Asn Val Leu 
        355                 360                 365 

Ile Gly Asp Met Ser Leu Val Gly Thr Arg Pro Pro Thr Val Asp Glu 
    370                 375                 380 

Phe Glu Lys Tyr Thr Pro Gly Gln Lys Arg Arg Leu Ser Phe Lys Pro 
385                 390                 395                 400 

Gly Ile Thr Gly Leu Trp Gln Val Ser Gly Arg Ser Asn Ile Thr Asp 
                405                 410                 415 

Phe Asp Asp Val Val Arg Leu Asp Leu Ala Tyr Ile Asp Asn Trp Thr 
            420                 425                 430 

Ile Trp Ser Asp Ile Lys Ile Leu Leu Lys Thr Val Lys Val Val Leu 
        435                 440                 445 

Leu Arg Glu Gly Ser Lys 
    450 

 
           
             31  
             149  
             PRT  
             Streptococcus suis  
             
               misc_feature  
               CPS1F  
             
           
            31 

Met Lys Val Cys Leu Val Gly Ser Ser Gly Gly His Leu Thr His Leu 
1               5                   10                  15 

Tyr Leu Leu Lys Pro Phe Trp Lys Glu Glu Glu Arg Phe Trp Val Thr 
            20                  25                  30 

Phe Asp Lys Glu Asp Ala Arg Ser Leu Leu Lys Asn Glu Lys Met Tyr 
        35                  40                  45 

Pro Cys Tyr Phe Pro Thr Asn Arg Asn Leu Ile Asn Leu Val Lys Asn 
    50                  55                  60 

Thr Phe Leu Ala Phe Lys Ile Leu Arg Asp Glu Lys Pro Asp Val Ile 
65                  70                  75                  80 

Ile Ser Ser Gly Ala Ala Val Ala Val Pro Phe Phe Tyr Ile Gly Lys 
                85                  90                  95 

Leu Phe Gly Ala Lys Thr Ile Tyr Ile Glu Val Phe Asp Arg Val Asn 
            100                 105                 110 

Lys Ser Thr Leu Thr Gly Lys Leu Val Tyr Pro Val Thr Asp Ile Phe 
        115                 120                 125 

Ile Val Gln Trp Glu Glu Met Lys Lys Val Tyr Pro Lys Ser Ile Asn 
    130                 135                 140 

Leu Gly Ser Ile Phe 
145 

 
           
             32  
             164  
             PRT  
             Streptococcus suis  
             
               misc_feature  
               CPS1G  
             
           
            32 

Met Ile Phe Val Thr Val Gly Thr His Glu Gln Gln Phe Asn Arg Leu 
1               5                   10                  15 

Ile Lys Glu Ile Asp Leu Leu Lys Lys Asn Gly Ser Ile Thr Asp Glu 
            20                  25                  30 

Ile Phe Ile Gln Thr Gly Tyr Ser Asp Tyr Ile Pro Glu Tyr Cys Lys 
        35                  40                  45 

Tyr Lys Lys Phe Leu Ser Tyr Lys Glu Met Glu Gln Tyr Ile Asn Lys 
    50                  55                  60 

Ser Glu Val Val Ile Cys His Gly Gly Pro Ala Thr Phe Met Asn Ser 
65                  70                  75                  80 

Leu Ser Lys Gly Lys Lys Gln Leu Leu Phe Pro Arg Gln Lys Lys Tyr 
                85                  90                  95 

Gly Glu His Val Asn Asp His Gln Val Glu Phe Val Arg Arg Ile Leu 
            100                 105                 110 

Gln Asp Asn Asn Ile Leu Phe Ile Glu Asn Ile Asp Asp Leu Phe Glu 
        115                 120                 125 

Lys Ile Ile Glu Val Ser Lys Gln Thr Asn Phe Thr Ser Asn Asn Asn 
    130                 135                 140 

Phe Phe Cys Glu Arg Leu Lys Gln Ile Val Glu Lys Phe Asn Glu Asp 
145                 150                 155                 160 

Gln Glu Asn Glu 

 
           
             33  
             388  
             PRT  
             Streptococcus suis  
             
               misc_feature  
               CPS1H  
             
           
            33 

Met Phe Lys Leu Phe Lys Tyr Asp Pro Glu Tyr Phe Ile Phe Lys Tyr 
1               5                   10                  15 

Phe Trp Leu Ile Ile Phe Ile Pro Glu Gln Lys Tyr Val Phe Leu Leu 
            20                  25                  30 

Ile Phe Met Asn Leu Ile Leu Phe His Ile Lys Phe Leu Lys Thr Lys 
        35                  40                  45 

Leu Ile Leu Lys Asn Glu Ile Leu Leu Phe Leu Leu Trp Ser Ile Leu 
    50                  55                  60 

Cys Phe Val Ser Val Val Thr Ser Met Phe Val Glu Ile Asn Phe Glu 
65                  70                  75                  80 

Arg Leu Phe Ala Asp Phe Thr Ala Pro Ile Ile Trp Ile Ile Ala Ile 
                85                  90                  95 

Met Tyr Tyr Asn Leu Tyr Ser Phe Ile Asn Ile Asp Tyr Lys Lys Leu 
            100                 105                 110 

Lys Asn Ser Ile Phe Phe Ser Phe Leu Val Leu Leu Gly Ile Ser Ala 
        115                 120                 125 

Leu Tyr Ile Ile Gln Asn Gly Lys Asp Ile Val Phe Leu Asp Arg His 
    130                 135                 140 

Leu Ile Gly Leu Asp Tyr Leu Ile Thr Gly Val Lys Thr Arg Leu Val 
145                 150                 155                 160 

Gly Phe Met Asn Tyr Pro Thr Leu Asn Thr Thr Thr Ile Ile Val Ser 
                165                 170                 175 

Ile Pro Leu Ile Phe Ala Leu Ile Lys Asn Lys Met Gln Gln Phe Phe 
            180                 185                 190 

Phe Leu Cys Leu Ala Phe Ile Pro Ile Tyr Leu Ser Gly Ser Arg Ile 
        195                 200                 205 

Gly Ser Leu Ser Leu Ala Ile Leu Ile Ile Cys Leu Leu Trp Arg Tyr 
    210                 215                 220 

Ile Gly Gly Lys Phe Ala Trp Ile Lys Lys Leu Ile Val Ile Phe Val 
225                 230                 235                 240 

Ile Leu Leu Ile Ile Leu Asn Thr Glu Leu Leu Tyr His Glu Ile Leu 
                245                 250                 255 

Ala Val Tyr Asn Ser Arg Glu Ser Ser Asn Glu Ala Arg Phe Ile Ile 
            260                 265                 270 

Tyr Gln Gly Ser Ile Asp Lys Val Leu Glu Asn Asn Ile Leu Phe Gly 
        275                 280                 285 

Tyr Gly Ile Ser Glu Tyr Ser Val Thr Gly Thr Trp Leu Gly Ser His 
    290                 295                 300 

Ser Gly Tyr Ile Ser Phe Phe Tyr Lys Ser Gly Ile Val Gly Leu Ile 
305                 310                 315                 320 

Leu Leu Met Phe Ser Phe Phe Tyr Val Ile Lys Lys Ser Tyr Gly Val 
                325                 330                 335 

Asn Gly Glu Thr Ala Leu Phe Tyr Phe Thr Ser Leu Ala Ile Phe Phe 
            340                 345                 350 

Ile Tyr Glu Thr Ile Asp Pro Ile Ile Ile Ile Leu Val Leu Phe Phe 
        355                 360                 365 

Ser Ser Ile Gly Ile Trp Asn Asn Ile Asn Phe Lys Lys Asp Met Glu 
    370                 375                 380 

Thr Lys Asn Glu 
385 

 
           
             34  
             322  
             PRT  
             Streptococcus suis  
             
               misc_feature  
               CPS1I  
             
           
            34 

Met Asn Asp Leu Ile Ser Val Ile Val Pro Ile Tyr Asn Val Gln Asp 
1               5                   10                  15 

Tyr Leu Asp Lys Cys Ile Asn Ser Ile Ile Asn Gln Thr Tyr Thr Asn 
            20                  25                  30 

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

Ile Cys Leu Asn Tyr Met Lys Asn Asp Gly Arg Ile Lys Tyr Tyr Lys 
    50                  55                  60 

Lys Ile Asn Gly Gly Leu Ala Asp Ala Arg Asn Phe Gly Leu Glu His 
65                  70                  75                  80 

Ala Thr Gly Lys Tyr Ile Ala Phe Val Asp Ser Asp Asp Tyr Ile Glu 
                85                  90                  95 

Val Ala Met Phe Glu Arg Met His Asp Asn Ile Thr Glu Tyr Asn Ala 
            100                 105                 110 

Asp Ile Ala Glu Ile Asp Phe Cys Leu Val Asp Glu Asn Gly Tyr Thr 
        115                 120                 125 

Lys Lys Lys Arg Asn Ser Asn Phe His Val Leu Thr Arg Glu Glu Thr 
    130                 135                 140 

Val Lys Glu Phe Leu Ser Gly Ser Asn Ile Glu Asn Asn Val Trp Cys 
145                 150                 155                 160 

Lys Leu Tyr Ser Arg Asp Ile Ile Lys Asp Ile Lys Phe Gln Ile Asn 
                165                 170                 175 

Asn Arg Ser Ile Gly Glu Asp Leu Leu Phe Asn Leu Glu Val Leu Asn 
            180                 185                 190 

Asn Val Thr Arg Val Val Val Asp Thr Arg Glu Tyr Tyr Tyr Asn Tyr 
        195                 200                 205 

Val Ile Arg Asn Ser Ser Leu Ile Asn Gln Lys Phe Ser Ile Asn Asn 
    210                 215                 220 

Ile Asp Leu Val Thr Arg Leu Glu Asn Tyr Pro Phe Lys Leu Lys Arg 
225                 230                 235                 240 

Glu Phe Ser His Tyr Phe Asp Ala Lys Val Ile Lys Glu Lys Val Lys 
                245                 250                 255 

Cys Leu Asn Lys Met Tyr Ser Thr Asp Cys Leu Asp Asn Glu Phe Leu 
            260                 265                 270 

Pro Ile Leu Glu Ser Tyr Arg Lys Glu Ile Arg Arg Tyr Pro Phe Ile 
        275                 280                 285 

Lys Ala Lys Arg Tyr Leu Ser Arg Lys His Leu Val Thr Leu Tyr Leu 
    290                 295                 300 

Met Lys Phe Ser Pro Lys Leu Tyr Val Met Leu Tyr Lys Lys Phe Gln 
305                 310                 315                 320 

Lys Gln 

 
           
             35  
             322  
             PRT  
             Streptococcus suis  
             
               misc_feature  
               CPS1J  
             
           
            35 

Met Asp Lys Ile Ser Val Ile Val Pro Val Tyr Asn Val Asp Lys Tyr 
1               5                   10                  15 

Leu Ser Ser Cys Ile Glu Ser Ile Ile Asn Gln Asn Tyr Lys Asn Ile 
            20                  25                  30 

Glu Ile Leu Leu Ile Asp Asp Gly Ser Val Asp Asp Ser Ala Lys Ile 
        35                  40                  45 

Cys Lys Glu Tyr Glu Lys Asp Lys Arg Val Lys Ile Phe Phe Thr Asn 
    50                  55                  60 

His Ser Gly Val Ser Asn Ala Arg Asn His Gly Ile Lys Arg Ser Thr 
65                  70                  75                  80 

Ala Glu Tyr Ile Met Phe Val Asp Ser Asp Asp Val Val Asp Ser Arg 
                85                  90                  95 

Leu Val Glu Lys Leu Tyr Phe Asn Ile Ile Lys Ser Arg Ser Asp Leu 
            100                 105                 110 

Ser Gly Cys Leu Tyr Ala Thr Phe Ser Glu Asn Ile Asn Asn Phe Glu 
        115                 120                 125 

Val Asn Asn Pro Asn Ile Asp Phe Glu Ala Ile Asn Thr Val Gln Asp 
    130                 135                 140 

Met Gly Glu Lys Asn Phe Met Asn Leu Tyr Ile Asn Asn Ile Phe Ser 
145                 150                 155                 160 

Thr Pro Val Cys Lys Leu Tyr Lys Lys Arg Tyr Ile Thr Asp Leu Phe 
                165                 170                 175 

Gln Glu Asn Gln Trp Leu Gly Glu Asp Leu Leu Phe Asn Leu His Tyr 
            180                 185                 190 

Leu Lys Asn Ile Asp Arg Val Ser Tyr Leu Thr Glu His Leu Tyr Phe 
        195                 200                 205 

Tyr Arg Arg Gly Ile Leu Ser Thr Val Asn Ser Phe Lys Glu Gly Val 
    210                 215                 220 

Phe Leu Gln Leu Glu Asn Leu Gln Lys Gln Val Ile Val Leu Phe Lys 
225                 230                 235                 240 

Gln Ile Tyr Gly Glu Asp Phe Asp Val Ser Ile Val Lys Asp Thr Ile 
                245                 250                 255 

Arg Trp Gln Val Phe Tyr Tyr Ser Leu Leu Met Phe Lys Tyr Gly Lys 
            260                 265                 270 

Gln Ser Ile Phe Asp Lys Phe Leu Ile Phe Arg Asn Leu Tyr Lys Lys 
        275                 280                 285 

Tyr Tyr Phe Asn Leu Leu Lys Val Ser Asn Lys Asn Ser Leu Ser Lys 
    290                 295                 300 

Asn Phe Cys Ile Arg Ile Val Ser Asn Lys Val Phe Lys Lys Ile Leu 
305                 310                 315                 320 

Trp Leu 

 
           
             36  
             278  
             PRT  
             Streptococcus suis  
             
               misc_feature  
               CPS1K  
             
           
            36 

Met Asp Thr Ile Ser Lys Ile Ser Ile Ile Val Pro Ile Tyr Asn Val 
1               5                   10                  15 

Glu Lys Tyr Leu Ser Lys Cys Ile Asp Ser Ile Val Asn Gln Thr Tyr 
            20                  25                  30 

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

Glu Glu Ile Cys Leu Ala Tyr Ala Lys Lys Asp Ser Arg Ile Arg Tyr 
    50                  55                  60 

Phe Lys Lys Glu Asn Gly Gly Leu Ser Asp Ala Arg Asn Tyr Gly Ile 
65                  70                  75                  80 

Ser Arg Ala Lys Gly Asp Tyr Leu Ala Phe Ile Asp Ser Asp Asp Phe 
                85                  90                  95 

Ile His Ser Glu Phe Ile Gln Arg Leu His Glu Ala Ile Glu Arg Glu 
            100                 105                 110 

Asn Ala Leu Val Ala Val Ala Gly Tyr Asp Arg Val Asp Ala Ser Gly 
        115                 120                 125 

His Phe Leu Thr Ala Glu Pro Leu Pro Thr Asn Gln Ala Val Leu Ser 
    130                 135                 140 

Gly Arg Asn Val Cys Lys Lys Leu Leu Glu Ala Asp Gly His Arg Phe 
145                 150                 155                 160 

Val Val Ala Cys Asn Lys Leu Tyr Lys Lys Glu Leu Phe Glu Asp Phe 
                165                 170                 175 

Arg Phe Glu Lys Gly Lys Ile His Glu Asp Glu Tyr Phe Thr Tyr Arg 
            180                 185                 190 

Leu Leu Tyr Glu Leu Glu Lys Val Ala Ile Val Lys Glu Cys Leu Tyr 
        195                 200                 205 

Tyr Tyr Val Asp Arg Glu Asn Ser Ile Thr Thr Ser Ser Met Thr Asp 
    210                 215                 220 

His Arg Phe His Cys Leu Leu Glu Phe Gln Asn Glu Arg Met Asp Phe 
225                 230                 235                 240 

Tyr Glu Ser Arg Gly Asp Lys Glu Leu Leu Leu Glu Cys Tyr Arg Ser 
                245                 250                 255 

Phe Leu Ala Phe Ala Val Leu Phe Leu Gly Lys Tyr Asn His Trp Leu 
            260                 265                 270 

Ser Lys Gln Gln Lys Lys 
        275 

 
           
             37  
             4519  
             DNA  
             Streptococcus suis  
             
               misc_feature  
               CPS9  
             
           
            37 

aagcttatcg tcaaggtgtt cgctatatcg tggcgacatc tcatagacga aaagggatgt     60 

ttgaaacacc agaaaaagtt atcatgacta actttcttca atttaaagac gcagtagcag    120 

aagtttatcc tgaaatacga ttgtgctatg gtgctgaatt gtattatagt aaagatatat    180 

taagcaaact tgaaaaaaag aaagtaccca cacttaatgg ctcgcgctat attcttttgg    240 

agttcagtag tgatactcct tggaaagaga ttcaagaagc agtgaacgaa gtgacgctac    300 

ttgggctaac tcccgtactt gcccatatag aacgatatga cgccctagcg tttcatgcag    360 

agagagtaga agagttaatt gacaagggat gctatactca ggtaaatagt aatcatgtgc    420 

tgaagcccac tttaattggt gatcgagcaa aagaatttaa aaaacgtact cggtattttt    480 

tagagcagga tttagtacat tgtgttgcta gcgatatgca taatttatct agtagacctc    540 

cgtttatgag ggaggcttat aagttgctaa cagaggaatt tggcaaagat aaagcgaaag    600 

cgttgctaaa aaagaatcct cttatgctat taaaaaacca ggcgatttaa actggttact    660 

ctagattgtg gagagaaaaa tggatttagg aactgttact gataaactgt tagaacgcaa    720 

cagtaaacga ttgatactcg tgtgcatgga tacgtgtctt cttatagttt ccatgatttt    780 

gagcagactg tttttggatg ttattattga cataccagat gaacgcttca ttcttgcagt    840 

tttattcgta tcaattttat atttgattct atcgtttaga ttaaaagtct tttcattaat    900 

tacgcgttac acagggtatc agagttatgt aaaaatagga cttagtttaa tatctgcgca    960 

ttcattgttt ttaattatct caatggtgtt gtggcaggct tttagttatc gtttcatctt   1020 

agtatcctta tttttgtcgt atgtaatgct cattactccg aggattgttt ggaaagtctt   1080 

acatgagacg agaaaaaatg ctatccgtaa gaaggatagc ccactaagaa tcttagtagt   1140 

aggtgctgga gatggtggta atatttttat caatactgtc aaagatcgaa aattgaattt   1200 

tgaaattgtc ggtatcgttg atcgtgatcc aaataaactt ggaacattta tccgtacggc   1260 

taaagtttta ggaaaccgta atgatattcc acgactggta gaggaattag ctgttgacca   1320 

agtgacgatt gccatccctt ctttaaatgg taaggagcga gagaagattg ttgaaatctg   1380 

taacactaca ggagtgaccg tcaataatat gccgagtatt gaagacatta tggcggggaa   1440 

catgtctgtc agtgcctttc aggaaattga cgtagcagac cttcttggtc gaccagaggt   1500 

tgttttggat caggatgaat tgaatcagtt tttccaaggg aaaacaatcc ttgtcacagg   1560 

agcaggtggc tctatcggtt cagagctatg tcgtcaaatt gctaagttta cgcctaaacg   1620 

cttgttgttg cttggacatg gagaaaattc aatctatctc attcatcgag agttactgga   1680 

aaagtaccaa ggtaagattg agttggtccc tctcattgca gatattcaag atagagaatt   1740 

gatttttagc ataatggctg aatatcaacc cgatgttgtt tatcatgctg cagcacataa   1800 

gcatgttcct ttgatggaat ataatccaca tgaagcagtg aagaataata tttttggaac   1860 

gaagaatgtg gctgaggcgg ctaaaactgc aaaggttgcc aaatttgtta tggtttcaac   1920 

agataaagct gttaatccac caaatgtcat gggagcgact aaacgtgttg cagaaatgat   1980 

tgttacaggt ttaaacgagc caggtcagac tcaatttgcg gcagtccggt ttgggaatgt   2040 

tctaggtagt cgtggaagtg ttgttccgct attcaaagag caaattagaa aaggtggacc   2100 

tgttacggtt accgacttta ggatgactcg ttatttcatg acgattcctg aggcaagtcg   2160 

tttggttatc caagctggac atttggcaaa aggtggagaa atatttgtct tggatatggg   2220 

cgagccagta caaatcctgg aattggcaag aaaagttatc ttgttaagtg gacacacaga   2280 

ggaagaaatc gggattgtag aatctggaat cagaccaggc gagaaactct acgaggaatt   2340 

attatcaaca gaagaacgtg tcagcgaaca gattcatgaa aaaatatttg tgggtcgcgt   2400 

tacaaataag cagtcggaca ttgtcaattc atttatcaat ggattactcc aaaaagatag   2460 

aaatgaatta aaaaatatgt tgattgaatt tgcaaaacaa gaataagaaa gtaaaaaata   2520 

tttttacttt cctagagttt aaacgatgtt taagttctag gaaggttaga atacctaatt   2580 

aacaacaata ttactattta ttaagagtca gataatagca actaagtgct acaaactatc   2640 

tttataataa gtatatttgg tcaaaaggga gatgtgaaat gtatccaatt tgtaaacgta   2700 

ttttagcaat tattatctca gggattgcta ttgttgttct gagtccaatt ttattattga   2760 

ttgcattggc aattaaatta gattctaaag gtccggtatt atttaaacaa aagcgggttg   2820 

gtaaaaacaa gtcatacttt atgatttata aattccgttc tatgtacgtt gacgcaccaa   2880 

gtgatatgcc gactcatcta ttaaaggatc ctaaggcgat gattaccaag gtgggcgcgt   2940 

ttctcagaaa aacaagttta gatgaactgc cacagctttt taatattttt aaaggtgaaa   3000 

tggcgattgt tggtccacgc ccagccttat ggaatcaata tgacttaatt gaagagcgag   3060 

ataaatatgg tgcaaatgat attcgtcctg gactaaccgg ttgggctcaa attaatggtc   3120 

gtgatgaatt ggaaattgat gaaaagtcaa aattagatgg atattatgtt caaaatatga   3180 

gtctaggttt ggatattaaa tgtttcttag gtacattcct cagtgtagcc agaagcgaag   3240 

gtgttgttga aggtggaaca gggcagaaag gaaaaggatg aaattttcag tattaatgtc   3300 

ggtctatgag aaagaaaaac cagagtttct tagggaatct ttggaaagca tccttgtcaa   3360 

tcaaacaatg attccaacgg aggttgtctt ggtagaggat gggccactca atcagagctt   3420 

atatagtatt ttagaagaat ttaaaagtcg attttcattt tttaaaacga tagccttgga   3480 

aaagaattcg ggtttaggaa ttgcactgaa tgaaggtttg aaacattgta attatgagtg   3540 

ggtttgcacg aaatggattc tgatgatgtt gcatatacat acacgttttg aaaagcaagt   3600 

taactttata aaacaaaacc cgactataga tattgagata gatgagttct taaattctac   3660 

tagtgaaata gtttctcata aaaatgttcc aacccagcac gatgaaatat taaagatggc   3720 

aaggcgggag aaatccatgt gccacatgac tgtaatgttt aaaaagaaaa gtgtcgagag   3780 

agcagggggg tatcaaacac ttccgtacgt agaagattat ttcctttggg tgcgcatgat   3840 

tgcttcagga tcgaaatttg caaacattga tgaaacacta gttcttgcac gtgttggaaa   3900 

tgggatgttc aataggaggg ggaacagaga acaaattaac agttggacat tactaattga   3960 

atttatgtta gctcaaggaa ttgttacacc actagatgta tttattaatc aaatttacat   4020 

tagggtcttt gtttatatgc caacttggat aaagaaactc atttatggaa aaatcttaag   4080 

gaaatagtat gattacagta ttgatggcta catataatgg aagcccattt ataataaaac   4140 

agttagattc aattcgaaat caaagtgtat cagcagacaa agttattatt tgggatgatt   4200 

gctcgacaga tgatacaata aaaataataa aagattatat aaaaaaatat tctttggatt   4260 

catgggttgt ctctcaaaat aaatctaatc aggggcatta tcaaacattt ataaatttga   4320 

caaagttagt tcaggaagga atagtctttt tttcagatca agatgatatt tgggactgtc   4380 

ataaaattga gacaatgctt ccaatctttg acagagaaaa tgtatcaatg gtgttttgca   4440 

aatccagatt gattgatgaa aacggaaata ttatcagtag cccagatact tcggatagaa   4500 

tcaatacgta ctctctaga                                                4519 

 
           
             38  
             215  
             PRT  
             Streptococcus suis  
             
               misc_feature  
               CPS9D  
             
           
            38 

Ala Tyr Arg Gln Gly Val Arg Tyr Ile Val Ala Thr Ser His Arg Arg 
1               5                   10                  15 

Lys Gly Met Phe Glu Thr Pro Glu Lys Val Ile Met Thr Asn Phe Leu 
            20                  25                  30 

Gln Phe Lys Asp Ala Val Ala Glu Val Tyr Pro Glu Ile Arg Leu Cys 
        35                  40                  45 

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

Lys Lys Lys Val Pro Thr Leu Asn Gly Ser Arg Tyr Ile Leu Leu Glu 
65                  70                  75                  80 

Phe Ser Ser Asp Thr Pro Trp Lys Glu Ile Gln Glu Ala Val Asn Glu 
                85                  90                  95 

Val Thr Leu Leu Gly Leu Thr Pro Val Leu Ala His Ile Glu Arg Tyr 
            100                 105                 110 

Asp Ala Leu Ala Phe His Ala Glu Arg Val Glu Glu Leu Ile Asp Lys 
        115                 120                 125 

Gly Cys Tyr Thr Gln Val Asn Ser Asn His Val Leu Lys Pro Thr Leu 
    130                 135                 140 

Ile Gly Asp Arg Ala Lys Glu Phe Lys Lys Arg Thr Arg Tyr Phe Leu 
145                 150                 155                 160 

Glu Gln Asp Leu Val His Cys Val Ala Ser Asp Met His Asn Leu Ser 
                165                 170                 175 

Ser Arg Pro Pro Phe Met Arg Glu Ala Tyr Lys Leu Leu Thr Glu Glu 
            180                 185                 190 

Phe Gly Lys Asp Lys Ala Lys Ala Leu Leu Lys Lys Asn Pro Leu Met 
        195                 200                 205 

Leu Leu Lys Asn Gln Ala Ile 
    210                 215 

 
           
             39  
             608  
             PRT  
             Streptococcus suis  
             
               misc_feature  
               CPS9E  
             
           
            39 

Met Asp Leu Gly Thr Val Thr Asp Lys Leu Leu Glu Arg Asn Ser Lys 
1               5                   10                  15 

Arg Leu Ile Leu Val Cys Met Asp Thr Cys Leu Leu Ile Val Ser Met 
            20                  25                  30 

Ile Leu Ser Arg Leu Phe Leu Asp Val Ile Ile Asp Ile Pro Asp Glu 
        35                  40                  45 

Arg Phe Ile Leu Ala Val Leu Phe Val Ser Ile Leu Tyr Leu Ile Leu 
    50                  55                  60 

Ser Phe Arg Leu Lys Val Phe Ser Leu Ile Thr Arg Tyr Thr Gly Tyr 
65                  70                  75                  80 

Gln Ser Tyr Val Lys Ile Gly Leu Ser Leu Ile Ser Ala His Ser Leu 
                85                  90                  95 

Phe Leu Ile Ile Ser Met Val Leu Trp Gln Ala Phe Ser Tyr Arg Phe 
            100                 105                 110 

Ile Leu Val Ser Leu Phe Leu Ser Tyr Val Met Leu Ile Thr Pro Arg 
        115                 120                 125 

Ile Val Trp Lys Val Leu His Glu Thr Arg Lys Asn Ala Ile Arg Lys 
    130                 135                 140 

Lys Asp Ser Pro Leu Arg Ile Leu Val Val Gly Ala Gly Asp Gly Gly 
145                 150                 155                 160 

Asn Ile Phe Ile Asn Thr Val Lys Asp Arg Lys Leu Asn Phe Glu Ile 
                165                 170                 175 

Val Gly Ile Val Asp Arg Asp Pro Asn Lys Leu Gly Thr Phe Ile Arg 
            180                 185                 190 

Thr Ala Lys Val Leu Gly Asn Arg Asn Asp Ile Pro Arg Leu Val Glu 
        195                 200                 205 

Glu Leu Ala Val Asp Gln Val Thr Ile Ala Ile Pro Ser Leu Asn Gly 
    210                 215                 220 

Lys Glu Arg Glu Lys Ile Val Glu Ile Cys Asn Thr Thr Gly Val Thr 
225                 230                 235                 240 

Val Asn Asn Met Pro Ser Ile Glu Asp Ile Met Ala Gly Asn Met Ser 
                245                 250                 255 

Val Ser Ala Phe Gln Glu Ile Asp Val Ala Asp Leu Leu Gly Arg Pro 
            260                 265                 270 

Glu Val Val Leu Asp Gln Asp Glu Leu Asn Gln Phe Phe Gln Gly Lys 
        275                 280                 285 

Thr Ile Leu Val Thr Gly Ala Gly Gly Ser Ile Gly Ser Glu Leu Cys 
    290                 295                 300 

Arg Gln Ile Ala Lys Phe Thr Pro Lys Arg Leu Leu Leu Leu Gly His 
305                 310                 315                 320 

Gly Glu Asn Ser Ile Tyr Leu Ile His Arg Glu Leu Leu Glu Lys Tyr 
                325                 330                 335 

Gln Gly Lys Ile Glu Leu Val Pro Leu Ile Ala Asp Ile Gln Asp Arg 
            340                 345                 350 

Glu Leu Ile Phe Ser Ile Met Ala Glu Tyr Gln Pro Asp Val Val Tyr 
        355                 360                 365 

His Ala Ala Ala His Lys His Val Pro Leu Met Glu Tyr Asn Pro His 
    370                 375                 380 

Glu Ala Val Lys Asn Asn Ile Phe Gly Thr Lys Asn Val Ala Glu Ala 
385                 390                 395                 400 

Ala Lys Thr Ala Lys Val Ala Lys Phe Val Met Val Ser Thr Asp Lys 
                405                 410                 415 

Ala Val Asn Pro Pro Asn Val Met Gly Ala Thr Lys Arg Val Ala Glu 
            420                 425                 430 

Met Ile Val Thr Gly Leu Asn Glu Pro Gly Gln Thr Gln Phe Ala Ala 
        435                 440                 445 

Val Arg Phe Gly Asn Val Leu Gly Ser Arg Gly Ser Val Val Pro Leu 
    450                 455                 460 

Phe Lys Glu Gln Ile Arg Lys Gly Gly Pro Val Thr Val Thr Asp Phe 
465                 470                 475                 480 

Arg Met Thr Arg Tyr Phe Met Thr Ile Pro Glu Ala Ser Arg Leu Val 
                485                 490                 495 

Ile Gln Ala Gly His Leu Ala Lys Gly Gly Glu Ile Phe Val Leu Asp 
            500                 505                 510 

Met Gly Glu Pro Val Gln Ile Leu Glu Leu Ala Arg Lys Val Ile Leu 
        515                 520                 525 

Leu Ser Gly His Thr Glu Glu Glu Ile Gly Ile Val Glu Ser Gly Ile 
    530                 535                 540 

Arg Pro Gly Glu Lys Leu Tyr Glu Glu Leu Leu Ser Thr Glu Glu Arg 
545                 550                 555                 560 

Val Ser Glu Gln Ile His Glu Lys Ile Phe Val Gly Arg Val Thr Asn 
                565                 570                 575 

Lys Gln Ser Asp Ile Val Asn Ser Phe Ile Asn Gly Leu Leu Gln Lys 
            580                 585                 590 

Asp Arg Asn Glu Leu Lys Asn Met Leu Ile Glu Phe Ala Lys Gln Glu 
        595                 600                 605 

 
           
             40  
             200  
             PRT  
             Streptococcus suis  
             
               misc_feature  
               CPS9F  
             
           
            40 

Met Tyr Pro Ile Cys Lys Arg Ile Leu Ala Ile Ile Ile Ser Gly Ile 
1               5                   10                  15 

Ala Ile Val Val Leu Ser Pro Ile Leu Leu Leu Ile Ala Leu Ala Ile 
            20                  25                  30 

Lys Leu Asp Ser Lys Gly Pro Val Leu Phe Lys Gln Lys Arg Val Gly 
        35                  40                  45 

Lys Asn Lys Ser Tyr Phe Met Ile Tyr Lys Phe Arg Ser Met Tyr Val 
    50                  55                  60 

Asp Ala Pro Ser Asp Met Pro Thr His Leu Leu Lys Asp Pro Lys Ala 
65                  70                  75                  80 

Met Ile Thr Lys Val Gly Ala Phe Leu Arg Lys Thr Ser Leu Asp Glu 
                85                  90                  95 

Leu Pro Gln Leu Phe Asn Ile Phe Lys Gly Glu Met Ala Ile Val Gly 
            100                 105                 110 

Pro Arg Pro Ala Leu Trp Asn Gln Tyr Asp Leu Ile Glu Glu Arg Asp 
        115                 120                 125 

Lys Tyr Gly Ala Asn Asp Ile Arg Pro Gly Leu Thr Gly Trp Ala Gln 
    130                 135                 140 

Ile Asn Gly Arg Asp Glu Leu Glu Ile Asp Glu Lys Ser Lys Leu Asp 
145                 150                 155                 160 

Gly Tyr Tyr Val Gln Asn Met Ser Leu Gly Leu Asp Ile Lys Cys Phe 
                165                 170                 175 

Leu Gly Thr Phe Leu Ser Val Ala Arg Ser Glu Gly Val Val Glu Gly 
            180                 185                 190 

Gly Thr Gly Gln Lys Gly Lys Gly 
        195                 200 

 
           
             41  
             269  
             PRT  
             Streptococcus suis  
             
               misc_feature  
               CPS2G  
             
           
            41 

Met Lys Phe Ser Val Leu Met Ser Val Tyr Glu Lys Glu Lys Pro Glu 
1               5                   10                  15 

Phe Leu Arg Glu Ser Leu Glu Ser Ile Leu Val Asn Gln Thr Met Ile 
            20                  25                  30 

Pro Thr Glu Val Val Leu Val Glu Asp Gly Pro Leu Asn Gln Ser Leu 
        35                  40                  45 

Tyr Ser Ile Leu Glu Glu Phe Lys Ser Arg Phe Ser Phe Phe Lys Thr 
    50                  55                  60 

Ile Ala Leu Glu Lys Asn Ser Gly Leu Gly Ile Ala Leu Asn Glu Gly 
65                  70                  75                  80 

Leu Lys His Cys Asn Tyr Glu Trp Val Cys Thr Lys Trp Ile Leu Met 
                85                  90                  95 

Met Leu His Ile His Thr Arg Phe Glu Lys Gln Val Asn Phe Ile Lys 
            100                 105                 110 

Gln Asn Pro Thr Ile Asp Ile Glu Ile Asp Glu Phe Leu Asn Ser Thr 
        115                 120                 125 

Ser Glu Ile Val Ser His Lys Asn Val Pro Thr Gln His Asp Glu Ile 
    130                 135                 140 

Leu Lys Met Ala Arg Arg Glu Lys Ser Met Cys His Met Thr Val Met 
145                 150                 155                 160 

Phe Lys Lys Lys Ser Val Glu Arg Ala Gly Gly Tyr Gln Thr Leu Pro 
                165                 170                 175 

Tyr Val Glu Asp Tyr Phe Leu Trp Val Arg Met Ile Ala Ser Gly Ser 
            180                 185                 190 

Lys Phe Ala Asn Ile Asp Glu Thr Leu Val Leu Ala Arg Val Gly Asn 
        195                 200                 205 

Gly Met Phe Asn Arg Arg Gly Asn Arg Glu Gln Ile Asn Ser Trp Thr 
    210                 215                 220 

Leu Leu Ile Glu Phe Met Leu Ala Gln Gly Ile Val Thr Pro Leu Asp 
225                 230                 235                 240 

Val Phe Ile Asn Gln Ile Tyr Ile Arg Val Phe Val Tyr Met Pro Thr 
                245                 250                 255 

Trp Ile Lys Lys Leu Ile Tyr Gly Lys Ile Leu Arg Lys 
            260                 265 

 
           
             42  
             143  
             PRT  
             Streptococcus suis  
             
               misc_feature  
               CPS9H  
             
           
            42 

Met Ile Thr Val Leu Met Ala Thr Tyr Asn Gly Ser Pro Phe Ile Ile 
1               5                   10                  15 

Lys Gln Leu Asp Ser Ile Arg Asn Gln Ser Val Ser Ala Asp Lys Val 
            20                  25                  30 

Ile Ile Trp Asp Asp Cys Ser Thr Asp Asp Thr Ile Lys Ile Ile Lys 
        35                  40                  45 

Asp Tyr Ile Lys Lys Tyr Ser Leu Asp Ser Trp Val Val Ser Gln Asn 
    50                  55                  60 

Lys Ser Asn Gln Gly His Tyr Gln Thr Phe Ile Asn Leu Thr Lys Leu 
65                  70                  75                  80 

Val Gln Glu Gly Ile Val Phe Phe Ser Asp Gln Asp Asp Ile Trp Asp 
                85                  90                  95 

Cys His Lys Ile Glu Thr Met Leu Pro Ile Phe Asp Arg Glu Asn Val 
            100                 105                 110 

Ser Met Val Phe Cys Lys Ser Arg Leu Ile Asp Glu Asn Gly Asn Ile 
        115                 120                 125 

Ile Ser Ser Pro Asp Thr Ser Asp Arg Ile Asn Thr Tyr Ser Leu 
    130                 135                 140 

 
           
             43  
             3738  
             DNA  
             Streptococcus suis  
             
               misc_feature  
               CPS7  
             
           
            43 

ctgcagcaca taagcatgtt ccattgatgg aatataatcc acatgaagca gtgaagaata     60 

atatttttgg aacgaagaat gtggctgagg cggctaaaac tgcaaaggtt gccaaatttg    120 

ttatggtttc aacagataaa gctgttaatc cgccaaatgt catgggagcg actaaacgtg    180 

ttgcagaaat gattgtaaca ggtttaaacg agccaggtca gactcaattt gcggcagtcc    240 

gttttgggaa tgttctaggt agtcgtggaa gtgttgttcc gctattcaaa gagcaaatta    300 

gaaaaggtgg acctgttacg gttaccgact ttaggatgac tcgttatttc atgacgattc    360 

ctgaggcaag tcgtttggtt atccaagctg gacatttggc aaaaggtgga gaaatctttg    420 

tcttggatat gggtgagcca gtacaaatcc tggaattggc aagaaaagtt atcttgttaa    480 

gcggacatac agaggaagaa atcgggattg tagaatctgg aatcagacca ggcgagaaac    540 

tctacgagga attgttatca acagaagaac gtgtcagcga acagattcat gaaaaaatat    600 

ttgtgggtcg cgttacaaat aagcagtcgg acattgtcaa ttcatttatc aatggattac    660 

tccaaaaaga tagaaatgaa ttaaaagata tgttgattga atttgcaaaa caagaataag    720 

aaagtaaaaa atatttttac tttcctagag tttaaacgat gtttaagttc taggaaggtt    780 

ggaattgctt tcgtggaggt gatagataga aacctatata tttgtagaag aaaggatatt    840 

aaactaaagg tgaatcggaa cataaagttt agatagagtt ggtatttaat gccaaacagg    900 

tgaatgcaac ctctcgctcg ttactaagca ggagatagta aagttgcttg aaagagagtt    960 

tgttaatcag tataagtagg ctaaagtgag aatatatatc tattattatc ggtaatgata   1020 

ctattattga gaattattgt agtggggata aaaataattt ttggtgattt tatcgtccga   1080 

cttaaaggtg ggttaaaaaa gtacttatat tcttttagaa ttgatgaaaa atatggggga   1140 

atataatatt tataggagat acgatgacta gagtagagtt gattactaga gaatttttta   1200 

agaagaatga agcaaccagt aaatattttc agaagataga atcaagaaga ggtgaattat   1260 

ttattaaatt ctttatggat aagttacttg cgcttatcct attattgcta ttatccccag   1320 

taatcattat attagctatt tggataaaat tagatagtaa ggggccaatt ttttatcgcc   1380 

aagaacgtgt tacgagatat ggtcgaattt ttagaatatt taagtttaga acaatgattt   1440 

ctgatgcgga taaagtcgga agtcttgtca cagtcggtca agataatcgt attacgaaag   1500 

tcggtcacat tatcagaaaa tatcggctgg acgaagtgcc ccaacttttt aatgttttaa   1560 

tgggggatat gagctttgta ggtgtaagac cagaagtaca aaaatatgta aatcagtata   1620 

ctgatgaaat gtttgcgacg ttacttttac ctgcaggaat tacttcacca gcgagtattg   1680 

catataagga tgaagatatt gttttagaag aatattgttc tcaaggctat agtcctgatg   1740 

aagcatatgt tcaaaaagta ttaccagaaa aaatgaagta caatttggaa tatatcagaa   1800 

actttggaat tatttctgat tttaaagtaa tgattgatac agtaattaaa gtaataaaat   1860 

aggagattaa aatgacaaaa agacaaaata ttccattttc accaccagat attacccaag   1920 

ctgaaattga tgaagttatt gacacactaa aatctggttg gattacaaca ggaccaaaga   1980 

caaaagagct agaacgtcgg ctatcagtat ttacaggaac caataaaact gtgtgtttaa   2040 

attctgctac tgcaggattg gaactagtct tacgaattct tggtgttgga cccggagatg   2100 

aagttattgt tcctgctatg acctatactg cctcatgtag tgtcattact catgtaggag   2160 

caactcctgt gatggttgat attcaaaaaa acagctttga gatggaatat gatgctttgg   2220 

aaaaagcgat tactccgaaa acaaaagtta tcattcctgt tgatctagct ggtattcctt   2280 

gtgattatga taagatttat accatcgtag aaaacaaacg ctctttgtat gttgcttctg   2340 

ataataaatg gcagaaactt tttgggcgag ttattatcct atctgatagt gcacactcac   2400 

taggtgctag ttataaggga aaaccagcgg gttccctagc agattttacc tcattttctt   2460 

tccatgcagt taagaatttt acaactgctg aaggaggtag tgtgacatgg agatcacatc   2520 

ctgatttgga tgacgaagag atgtataaag agtttcagat ttactctctt catggtcaga   2580 

caaaggatgc attagctaag acacaattag ggtcatggga atatgacatt gttattcctg   2640 

gttacaagtg taatatgaca gatattatgg caggtatcgg tcttgtgcaa ttagaacgtt   2700 

acccatcttt gttgaatcgt cgcagagaaa tcattgagaa atacaatgct ggctttgagg   2760 

ggacttcgat taagccgttg gtacacctga cggaagataa acaatcgtct atgcacttgt   2820 

atatcacgca tctacaaggc tatactttag aacaacgaaa tgaagtcatt caaaaaatgg   2880 

ctgaagcagg tattgcgtgc aatgttcact acaaaccatt acctcttctc acagcctaca   2940 

agaatcttgg ttttgaaatg aaagattttc cgaatgccta tcagtatttt gaaaatgaag   3000 

ttacactgcc tcttcatacc aacttgagtg atgaagatgt ggagtatgtg atagaaatgt   3060 

ttttaaaaat tgttagtaga gattagttat tttggaagga gatatggtgg aaagagatat   3120 

ggtggaaaga gacacgttgg tatctataat aatgccctcg tggaatacag ctaagtatat   3180 

atctgaatca atccagtcag tgttggacca aacacaccaa aattgggaac ttataatcgt   3240 

tgatgattgt tctaatgacg aaactgaaaa agttgtttcg catttcaaag attcaagaat   3300 

aaagtttttt aaaaattcga ataatttagg ggcagctcta acacgaaata aggcactaag   3360 

aaaagctaga ggtaggtgga ttgcgttctt ggattcagat gatttatggc acccgagtaa   3420 

gctagaaaaa cagcttgaat ttatgaaaaa taatggatat tcatttactt atcacaattt   3480 

tgaaaagatt gatgaatcta gtcagtcttt acgtgtcctg gtgtcaggac cagcaattgt   3540 

gactagaaaa atgatgtaca attacggcta tccagggtgt ttgactttca tgtatgatgc   3600 

agacaaaatg ggtttaattc agataaaaga tataaagaaa aataacgatt atgcgatatt   3660 

acttcaattg tgtaagaagt atgactgtta tcttttaaat gaaagtttag cttcgtatcg   3720 

aattagaaaa aaatcgat                                                 3738 

 
           
             44  
             238  
             PRT  
             Streptococcus suis  
             
               misc_feature  
               CPS7E  
             
           
            44 

Ala Ala His Lys His Val Pro Leu Met Glu Tyr Asn Pro His Glu Ala 
1               5                   10                  15 

Val Lys Asn Asn Ile Phe Gly Thr Lys Asn Val Ala Glu Ala Ala Lys 
            20                  25                  30 

Thr Ala Lys Val Ala Lys Phe Val Met Val Ser Thr Asp Lys Ala Val 
        35                  40                  45 

Asn Pro Pro Asn Val Met Gly Ala Thr Lys Arg Val Ala Glu Met Ile 
    50                  55                  60 

Val Thr Gly Leu Asn Glu Pro Gly Gln Thr Gln Phe Ala Ala Val Arg 
65                  70                  75                  80 

Phe Gly Asn Val Leu Gly Ser Arg Gly Ser Val Val Pro Leu Phe Lys 
                85                  90                  95 

Glu Gln Ile Arg Lys Gly Gly Pro Val Thr Val Thr Asp Phe Arg Met 
            100                 105                 110 

Thr Arg Tyr Phe Met Thr Ile Pro Glu Ala Ser Arg Leu Val Ile Gln 
        115                 120                 125 

Ala Gly His Leu Ala Lys Gly Gly Glu Ile Phe Val Leu Asp Met Gly 
    130                 135                 140 

Glu Pro Val Gln Ile Leu Glu Leu Ala Arg Lys Val Ile Leu Leu Ser 
145                 150                 155                 160 

Gly His Thr Glu Glu Glu Ile Gly Ile Val Glu Ser Gly Ile Arg Pro 
                165                 170                 175 

Gly Glu Lys Leu Tyr Glu Glu Leu Leu Ser Thr Glu Glu Arg Val Ser 
            180                 185                 190 

Glu Gln Ile His Glu Lys Ile Phe Val Gly Arg Val Thr Asn Lys Gln 
        195                 200                 205 

Ser Asp Ile Val Asn Ser Phe Ile Asn Gly Leu Leu Gln Lys Asp Arg 
    210                 215                 220 

Asn Glu Leu Lys Asp Met Leu Ile Glu Phe Ala Lys Gln Glu 
225                 230                 235 

 
           
             45  
             232  
             PRT  
             Streptococcus suis  
             
               misc_feature  
               CPS7F  
             
           
            45 

Met Thr Arg Val Glu Leu Ile Thr Arg Glu Phe Phe Lys Lys Asn Glu 
1               5                   10                  15 

Ala Thr Ser Lys Tyr Phe Gln Lys Ile Glu Ser Arg Arg Gly Glu Leu 
            20                  25                  30 

Phe Ile Lys Phe Phe Met Asp Lys Leu Leu Ala Leu Ile Leu Leu Leu 
        35                  40                  45 

Leu Leu Ser Pro Val Ile Ile Ile Leu Ala Ile Trp Ile Lys Leu Asp 
    50                  55                  60 

Ser Lys Gly Pro Ile Phe Tyr Arg Gln Glu Arg Val Thr Arg Tyr Gly 
65                  70                  75                  80 

Arg Ile Phe Arg Ile Phe Lys Phe Arg Thr Met Ile Ser Asp Ala Asp 
                85                  90                  95 

Lys Val Gly Ser Leu Val Thr Val Gly Gln Asp Asn Arg Ile Thr Lys 
            100                 105                 110 

Val Gly His Ile Ile Arg Lys Tyr Arg Leu Asp Glu Val Pro Gln Leu 
        115                 120                 125 

Phe Asn Val Leu Met Gly Asp Met Ser Phe Val Gly Val Arg Pro Glu 
    130                 135                 140 

Val Gln Lys Tyr Val Asn Gln Tyr Thr Asp Glu Met Phe Ala Thr Leu 
145                 150                 155                 160 

Leu Leu Pro Ala Gly Ile Thr Ser Pro Ala Ser Ile Ala Tyr Lys Asp 
                165                 170                 175 

Glu Asp Ile Val Leu Glu Glu Tyr Cys Ser Gln Gly Tyr Ser Pro Asp 
            180                 185                 190 

Glu Ala Tyr Val Gln Lys Val Leu Pro Glu Lys Met Lys Tyr Asn Leu 
        195                 200                 205 

Glu Tyr Ile Arg Asn Phe Gly Ile Ile Ser Asp Phe Lys Val Met Ile 
    210                 215                 220 

Asp Thr Val Ile Lys Val Ile Lys 
225                 230 

 
           
             46  
             404  
             PRT  
             Streptococcus suis  
             
               misc_feature  
               CPS7G  
             
           
            46 

Met Thr Lys Arg Gln Asn Ile Pro Phe Ser Pro Pro Asp Ile Thr Gln 
1               5                   10                  15 

Ala Glu Ile Asp Glu Val Ile Asp Thr Leu Lys Ser Gly Trp Ile Thr 
            20                  25                  30 

Thr Gly Pro Lys Thr Lys Glu Leu Glu Arg Arg Leu Ser Val Phe Thr 
        35                  40                  45 

Gly Thr Asn Lys Thr Val Cys Leu Asn Ser Ala Thr Ala Gly Leu Glu 
    50                  55                  60 

Leu Val Leu Arg Ile Leu Gly Val Gly Pro Gly Asp Glu Val Ile Val 
65                  70                  75                  80 

Pro Ala Met Thr Tyr Thr Ala Ser Cys Ser Val Ile Thr His Val Gly 
                85                  90                  95 

Ala Thr Pro Val Met Val Asp Ile Gln Lys Asn Ser Phe Glu Met Glu 
            100                 105                 110 

Tyr Asp Ala Leu Glu Lys Ala Ile Thr Pro Lys Thr Lys Val Ile Ile 
        115                 120                 125 

Pro Val Asp Leu Ala Gly Ile Pro Cys Asp Tyr Asp Lys Ile Tyr Thr 
    130                 135                 140 

Ile Val Glu Asn Lys Arg Ser Leu Tyr Val Ala Ser Asp Asn Lys Trp 
145                 150                 155                 160 

Gln Lys Leu Phe Gly Arg Val Ile Ile Leu Ser Asp Ser Ala His Ser 
                165                 170                 175 

Leu Gly Ala Ser Tyr Lys Gly Lys Pro Ala Gly Ser Leu Ala Asp Phe 
            180                 185                 190 

Thr Ser Phe Ser Phe His Ala Val Lys Asn Phe Thr Thr Ala Glu Gly 
        195                 200                 205 

Gly Ser Val Thr Trp Arg Ser His Pro Asp Leu Asp Asp Glu Glu Met 
    210                 215                 220 

Tyr Lys Glu Phe Gln Ile Tyr Ser Leu His Gly Gln Thr Lys Asp Ala 
225                 230                 235                 240 

Leu Ala Lys Thr Gln Leu Gly Ser Trp Glu Tyr Asp Ile Val Ile Pro 
                245                 250                 255 

Gly Tyr Lys Cys Asn Met Thr Asp Ile Met Ala Gly Ile Gly Leu Val 
            260                 265                 270 

Gln Leu Glu Arg Tyr Pro Ser Leu Leu Asn Arg Arg Arg Glu Ile Ile 
        275                 280                 285 

Glu Lys Tyr Asn Ala Gly Phe Glu Gly Thr Ser Ile Lys Pro Leu Val 
    290                 295                 300 

His Leu Thr Glu Asp Lys Gln Ser Ser Met His Leu Tyr Ile Thr His 
305                 310                 315                 320 

Leu Gln Gly Tyr Thr Leu Glu Gln Arg Asn Glu Val Ile Gln Lys Met 
                325                 330                 335 

Ala Glu Ala Gly Ile Ala Cys Asn Val His Tyr Lys Pro Leu Pro Leu 
            340                 345                 350 

Leu Thr Ala Tyr Lys Asn Leu Gly Phe Glu Met Lys Asp Phe Pro Asn 
        355                 360                 365 

Ala Tyr Gln Tyr Phe Glu Asn Glu Val Thr Leu Pro Leu His Thr Asn 
    370                 375                 380 

Leu Ser Asp Glu Asp Val Glu Tyr Val Ile Glu Met Phe Leu Lys Ile 
385                 390                 395                 400 

Val Ser Arg Asp 

 
           
             47  
             210  
             PRT  
             Streptococcus suis  
             
               misc_feature  
               CPS7H  
             
           
            47 

Met Val Glu Arg Asp Met Val Glu Arg Asp Thr Leu Val Ser Ile Ile 
1               5                   10                  15 

Met Pro Ser Trp Asn Thr Ala Lys Tyr Ile Ser Glu Ser Ile Gln Ser 
            20                  25                  30 

Val Leu Asp Gln Thr His Gln Asn Trp Glu Leu Ile Ile Val Asp Asp 
        35                  40                  45 

Cys Ser Asn Asp Glu Thr Glu Lys Val Val Ser His Phe Lys Asp Ser 
    50                  55                  60 

Arg Ile Lys Phe Phe Lys Asn Ser Asn Asn Leu Gly Ala Ala Leu Thr 
65                  70                  75                  80 

Arg Asn Lys Ala Leu Arg Lys Ala Arg Gly Arg Trp Ile Ala Phe Leu 
                85                  90                  95 

Asp Ser Asp Asp Leu Trp His Pro Ser Lys Leu Glu Lys Gln Leu Glu 
            100                 105                 110 

Phe Met Lys Asn Asn Gly Tyr Ser Phe Thr Tyr His Asn Phe Glu Lys 
        115                 120                 125 

Ile Asp Glu Ser Ser Gln Ser Leu Arg Val Leu Val Ser Gly Pro Ala 
    130                 135                 140 

Ile Val Thr Arg Lys Met Met Tyr Asn Tyr Gly Tyr Pro Gly Cys Leu 
145                 150                 155                 160 

Thr Phe Met Tyr Asp Ala Asp Lys Met Gly Leu Ile Gln Ile Lys Asp 
                165                 170                 175 

Ile Lys Lys Asn Asn Asp Tyr Ala Ile Leu Leu Gln Leu Cys Lys Lys 
            180                 185                 190 

Tyr Asp Cys Tyr Leu Leu Asn Glu Ser Leu Ala Ser Tyr Arg Ile Arg 
        195                 200                 205 

Lys Lys 
    210 

 
           
             48  
             101  
             DNA  
             Streptococcus suis  
             
               misc_feature  
               (1)..(101)  
               N may be any nucleotide  
             
           
            48 

aagggcacct ctataaactc ccaaaattgc gaatttggag ttacgaaagc cttgttaaat     60 

caancatttt aaattttaga aaattagttt ttagagctcc c                        101 

 
           
             49  
             101  
             DNA  
             Streptococcus suis  
             
               misc_feature  
               (1)..(101)  
               N may be any nucleotide  
             
           
            49 

ggcgccacct ctataaattc ccaaaattgc gaatttcgag ttacgaaagc cttgttaaat     60 

caancatctt aaattttaga aaattagttt ttagaggtcc c                        101 

 
           
             50  
             101  
             DNA  
             Streptococcus suis  
             
               misc_feature  
               100 base pair repeat between CPS2O and CPS2P  
             
           
            50 

aagggcacct ctataaactc ccaaaattgc gaatttcgag ttacgaaagc cttgttaaat     60 

caaacatttt aaattttaga aaattagttt ttagaggtcc c                        101 

 
           
             51  
             120  
             PRT  
             Streptococcus suis  
             
               misc_feature  
               N-terminal part of CPS2J  
             
           
            51 

Met Ala Lys Val Ser Ile Ile Val Pro Ile Phe Asn Thr Glu Lys Tyr 
1               5                   10                  15 

Leu Arg Glu Cys Leu Asp Ser Ile Ile Ser Gln Ser Tyr Thr Asn Leu 
            20                  25                  30 

Glu Ile Leu Leu Ile Asp Asp Gly Ser Ser Asp Ser Ser Thr Asp Ile 
        35                  40                  45 

Cys Leu Glu Tyr Ala Glu Gln Asp Gly Arg Ile Lys Leu Phe Arg Leu 
    50                  55                  60 

Pro Asn Gly Gly Val Ser Asn Ala Arg Asn Tyr Gly Ile Lys Asn Ser 
65                  70                  75                  80 

Thr Ala Asn Tyr Ile Met Phe Val Asp Ser Asp Asp Ile Val Asp Gly 
                85                  90                  95 

Asn Ile Val Glu Ser Leu Tyr Thr Cys Leu Lys Glu Asn Asp Ser Asp 
            100                 105                 110 

Leu Ser Gly Gly Leu Leu Ala Thr 
        115                 120 

 
           
             52  
             120  
             PRT  
             Streptococcus suis  
             
               misc_feature  
               N-terminal part of CPS2K  
             
           
            52 

Met Ile Asn Ile Ser Ile Ile Val Pro Ile Tyr Asn Val Glu Gln Tyr 
1               5                   10                  15 

Leu Ser Lys Cys Ile Asn Ser Ile Val Asn Gln Thr Tyr Lys His Ile 
            20                  25                  30 

Glu Leu Leu Val Asn Asp Gly Ser Ser Thr Asp Asn Ser Glu Glu Ile 
        35                  40                  45 

Cys Leu Ala Tyr Ala Lys Lys Asp Ser Arg Ile Arg Tyr Phe Lys Lys 
    50                  55                  60 

Glu Asn Gly Gly Leu Ser Asp Ala Arg Asn Tyr Gly Ile Ser Arg Ala 
65                  70                  75                  80 

Lys Gly Asp Tyr Leu Ala Phe Ile Asp Ser Asp Asp Phe Ile His Ser 
                85                  90                  95 

Glu Phe Ile Gln Arg Leu Xaa His Glu Ala Ile Glu Arg Glu Asn Ala 
            100                 105                 110 

Leu Xaa Xaa Val Ala Val Ala Gly 
        115                 120 

 
           
             53  
             419  
             PRT  
             Streptococcus suis  
             
               misc_feature  
               ORF2Y  
             
           
            53 

Met Lys Lys Tyr Gln Val Ile Ile Gln Asp Ile Leu Thr Gly Ile Glu 
1               5                   10                  15 

Glu His Arg Phe Lys Arg Gly Glu Lys Leu Pro Ser Ile Arg Gln Leu 
            20                  25                  30 

Arg Glu Gln Tyr His Cys Ser Lys Asp Thr Val Gln Lys Ala Met Leu 
        35                  40                  45 

Glu Leu Lys Tyr Gln Asn Lys Ile Tyr Ala Val Glu Lys Ser Gly Tyr 
    50                  55                  60 

Tyr Ile Leu Glu Asp Arg Asp Phe Gln Asp His Thr Cys Arg Ala Gln 
65                  70                  75                  80 

Ser Tyr Arg Leu Ser Arg Ile Thr Tyr Glu Asp Phe Arg Ile Cys Leu 
                85                  90                  95 

Lys Glu Ser Leu Ile Gly Arg Glu Asn Tyr Leu Phe Asn Tyr Tyr His 
            100                 105                 110 

Gln Gln Glu Gly Leu Ala Glu Leu Ile Ser Ser Val Gln Ser Leu Leu 
        115                 120                 125 

Met Asp Tyr His Val Tyr Thr Lys Lys Asp Gln Leu Val Ile Thr Ala 
    130                 135                 140 

Gly Ser Gln Gln Ala Leu Tyr Ile Leu Thr Gln Met Glu Thr Leu Ala 
145                 150                 155                 160 

Gly Lys Thr Glu Ile Leu Ile Glu Asn Pro Thr Tyr Ser Arg Met Ile 
                165                 170                 175 

Glu Leu Ile Arg His Gln Gly Ile Pro Tyr Gln Thr Ile Glu Arg Asn 
            180                 185                 190 

Leu Asp Gly Ile Asp Leu Glu Glu Leu Glu Ser Ile Phe Gln Thr Gly 
        195                 200                 205 

Lys Ile Lys Phe Phe Tyr Thr Ile Pro Arg Leu His Asn Pro Leu Gly 
    210                 215                 220 

Ser Thr Tyr Asp Ile Ala Thr Lys Thr Ala Ile Val Lys Leu Ala Lys 
225                 230                 235                 240 

Gln Tyr Asp Val Tyr Ile Ile Glu Asp Asp Tyr Leu Ala Asp Phe Asp 
                245                 250                 255 

Ser Ser His Ser Leu Pro Leu His Tyr Leu Asp Thr Asp Asn Arg Val 
            260                 265                 270 

Ile Tyr Ile Lys Ser Phe Thr Pro Thr Leu Phe Pro Ala Leu Arg Ile 
        275                 280                 285 

Gly Ala Ile Ser Leu Pro Asn Gln Leu Arg Asp Ile Phe Ile Lys His 
    290                 295                 300 

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

Ser Leu Tyr Ile Asp Asn Gly Met Phe Ala Arg Asn Thr Gln His Leu 
                325                 330                 335 

His His Ile Tyr His Ala Gln Trp Asn Lys Ile Lys Asp Cys Leu Glu 
            340                 345                 350 

Lys Tyr Ala Leu Asn Ile Pro Tyr Arg Ile Pro Lys Gly Ser Val Thr 
        355                 360                 365 

Phe Gln Leu Ser Lys Gly Ile Leu Ser Pro Ser Ile Gln His Met Phe 
    370                 375                 380 

Gly Lys Cys Tyr Tyr Phe Ser Gly Gln Lys Ala Asp Phe Leu Gln Ile 
385                 390                 395                 400 

Phe Phe Glu Gln Asp Phe Ala Asp Lys Leu Glu Gln Phe Val Arg Tyr 
                405                 410                 415 

Leu Asn Glu