Vaccine composition against Streptococcus suis infection

Abstract

The present invention describes a vaccine composition comprising an effective amount of at least one polypeptide selected from IdeSsuis, rIdeSsuis, analogs or fragments thereof, or polynucleotides encoding them. The vaccine composition is used for preventive, intermittent or therapeutic treatment of Streptococcus suis infection in pigs or humans.

Description

[0001] Divisional application

[0002] The present application is a divisional application of application number 201580032725.1, filed on May 29, 2015, and entitled “Vaccine composition against Streptococcus suis infection”. Technical Field

[0003] The present invention relates to vaccine compositions and their use in immunizing and protecting mammals, particularly pigs and humans, against Streptococcus suis. Background Art

[0004] Streptococcus suis (S.suis) colonizes the respiratory, digestive and reproductive tracts of pigs and is one of the most important pig pathogens, causing different lesions such as meningitis, septicemia, arthritis and endocarditis.

[0005] S. suis infections cause high production losses in the pig industry worldwide. Antibiotics are commonly used to treat S. suis infections. However, recurrent infections often occur, and the ongoing discussion on reducing antibiotic use emphasizes the need for alternative control measures. In Europe, there are no licensed vaccines available, but autologous vaccines are commonly used. The main disadvantage is the fact that these vaccines only protect against homologous serotypes. But S. suis is a very diverse organism, and different serotypes cause disease in piglets. Especially serotype 2 strains play an important role in the disease in piglets worldwide.

[0006] S. suis serotype 2 has been identified as causing meningitis in adults in Asia, but human-to-human transmission of S. suis has not been detected to date.

[0007] Typically, infection induces an early antigen-specific immunoglobulin M (IgM) response, leading to affinity maturation and isotype switching. In addition, IgM antibodies are present prior to infection, are naturally occurring, and are important in linking innate immunity to acquired immunity.

[0008] In pigs, IgM is particularly important as monomeric membrane IgM (mIgM) because it is the only B cell receptor that occurs, as IgD is absent in pigs. In addition, the synthesis of IgM in newborn piglets is much earlier than the synthesis of IgG and IgA. IgM in colostrum is essential for the defense against pathogens, which is carried out by complement-mediated killing. Therefore, IgM antibodies are important in the defense against different pathogens.

[0009] Various virulence or virulence-related factors of S. suis serotype 2 have been identified, and in the capsule, it is the only known essential virulence factor to date that protects the pathogen from phagocytosis. Many surface-associated and secreted proteins of S. suis serotype 2 exhibit the same or very similar functions as the homologous factors of other pathogenic streptococci. Various human or animal pathogens such as S. pyogenes, S. equi subsp. equi and S. equi subsp. zooepidemicus express specific IgG endopeptidases that are homologous to each other.

[0010] Surface-associated or secreted factors with unique functions of S. suis have been first described by Seele et al. ("Identification of a Novel Host-specific IgM Protease in Streptococcus suis." 2013; Journal of Bacteriology, 195: 930-940). Seele et al. showed that this IgM protease (called IdeSsuis) does not function as an IgG endopeptidase. The IgM protease degrades opsonized IgM on the surface of the bacteria, thereby improving the survival of S. suis in the blood of bacterin-primed piglets. IdeSsuis is highly specific for IgM and does not cleave IgG or IgA. However, Seele et al. did not mention the function of IdeSsuis, rIdeSsuis and their analogs and fragments as effective vaccines against S. suis infection. It is only assumed that neutralization of the IdeSsuis IgM protease activity can substantially improve the protective efficacy of bacterins or other future vaccines that induce opsonized antibodies. However, there is no disclosure that IdeSsuis and related proteins can be used as proprietary immunizing agents in vaccines against S. suis infection.

[0011] Baums et al., "Surface-associated and secreted factors of Streptococcus suis in epidemiology, pathogenesis and vaccine development, Animal Health Research Reviews, Volume 10, Issue 01, June 2009, pp 65-83" discloses bacterial factors (surface-associated and secreted bacterial factors) that are thought to contribute to the interaction of S. suis with host factors and cells. The factors are given with respect to: (i) their identification and characteristics, (ii) their distribution in S. suis and (iii) their importance for virulence, immune response and vaccination. This review highlights many challenging issues that remain to be addressed in the future.

[0012] The problem to be solved according to the present invention is to overcome the problems described in the art and to provide a new vaccine composition to immunize and protect mammals, in particular pigs and humans, against S. suis infection. Summary of the invention

[0013] According to the present invention, this problem is solved by providing a vaccine composition comprising an effective amount of at least one polypeptide or at least one vector selected from the group consisting of:

[0014] (a) a protein called IdeSsuis, an analog or a fragment thereof,

[0015] (b) a protein called rIdySsuis, an analog or a fragment thereof,

[0016] (c) a vector into which is inserted a polynucleotide encoding a protein IdeSsuis, an analog or a fragment thereof,

[0017] (d) a vector into which is inserted a polynucleotide encoding the protein rIdeSsuis, an analog or a fragment thereof, and

[0018] At least one pharmaceutical carrier, diluent or adjuvant.

[0019] Detailed description of the invention

[0020] The vaccine composition used in the present invention comprises at least one single polypeptide defined by (a) or (b), and a pharmaceutical carrier or diluent or adjuvant or a mixture thereof. In addition, the vaccine may also comprise at least one single carrier defined by (c) or (d), and a pharmaceutical carrier, diluent or adjuvant or a mixture thereof.

[0021] According to the present invention, a vaccine composition is preferred, wherein the IdeSsuis of (a) comprises:

[0022] (aa) SEQ ID NO: 1 amino acid sequence;

[0023] (ab) a fragment or analog of the amino acid sequence of SEQ ID NO: 1; or

[0024] (ac) Fragments of (aa) or (ab) having IgM protease activity.

[0025] According to the present invention, further preferred is a vaccine composition, wherein the rIdeSsuis of (b) comprises or consists of:

[0026] (ba) the amino acid sequence of SEQ ID NO: 2, 6 or 7;

[0027] (bb) a fragment or analog of the amino acid sequence of SEQ ID NO: 2, 6 or 7

[0028] (bc) an amino acid sequence lacking the amino acids at positions 1 to 34 in the amino acid sequence of SEQ ID NO: 1;

[0029] (bd) an amino acid sequence having at least 60% homology, preferably 70% homology, and most preferably 85% homology to the amino acid sequence of the protein IdeSsuis of SEQ ID NO: 1;

[0030] (be) a fragment of (ba) or (bb) or (bc) or (bd) having IgM protease activity; or

[0031] (bf) a fragment of (ba) or (bb) or (bc) or (bd) comprising or consisting of the amino acid sequence of SEQ ID NO: 5 or an amino acid sequence that is at least 95% homologous thereto.

[0032] The amino acid sequence of SEQ ID NO: 2 represents the sequence of SEQ ID NO: 1, however amino acids 1-34 (signal peptide) are missing and a HIS tag has been added. Note that SEQ ID NO: 1 is derived from a serotype 2 strain of S. suis.

[0033] The amino acid sequence of SEQ ID NO:6 represents the N-terminal sequence of SEQ ID NO:2.

[0034] SEQ ID NO: 7 (also referred to as antigen rIdeSsuisB2) contains the complete amino acid sequence of the mature IdeSsuis protein of S. suis serotype 7 strain, but with the addition of an N-terminal HIS tag. The IdeSsuis protein of S. suis serotype 7 strain differs in the C terminal half of the protein, as it lacks a sequence of 114 amino acids compared to SEQ ID NO: 1. Amino acids 80 to 414 of SEQ ID NO: 7 (the so-called highly conserved part of the Mac-1 domain) correspond to 97.9% of the sequence of SEQ ID NO: 5. The overall identity between SEQ ID NO: 7 and 1 is 96.4% (excluding the N-terminal HIS tag and the gap of 114 amino acids).

[0035] The term "fragment or analog" as used herein is defined as follows:

[0036] "Analogs" can be considered as amino acid sequences similar to the amino acid sequences disclosed above and showing a homology level of at least 60%, preferably 70%, and most preferably 85% with the original amino acid sequence (e.g., SEQ ID NO: 1, 2, 6 or 7). Higher degrees of homology, such as 95%, are also contemplated herein. Homology as used herein refers to identity. Thus, sequences may differ from each other based on substitutions, deletions, or insertions.

[0037] The degree of identity can be determined using the protein blast program with the blastp algorithm with default parameters, e.g., Expected Threshold: 10, Word Size: 3, Matrix: BLOMSUM62, Gap Cost: Existence: 11 Extension: 1, and Composition Adjustment: Conditional Composition Score Matrix Adjustment (BLAST is a registered trademark of the National Library of Medicine). This program can search a protein database using a protein query. Identity reports the exact match between the aligned query and database sequences.

[0038] Preferably, an amino acid "substitution" is the result of replacing one amino acid with another amino acid having similar structure and / or chemical properties, i.e., a conservative amino acid substitution. Amino acid substitutions can be made based on similarity in polarity, charge, solubility, hydrophobicity, hydrophilicity, and / or amphipathic properties of the residues involved. For example, non-polar (hydrophobic) amino acids include alanine, leucine, isoleucine, valine, proline, phenylalanine, tryptophan, and methionine; polar neutral amino acids include glycine, serine, threonine, cysteine, tyrosine, asparagine, and glutamine; positively charged (basic) amino acids include arginine, lysine, and histidine; and negatively charged (acidic) amino acids include aspartic acid and glutamic acid.

[0039] "Insertion" or "deletion" is usually within the range of about 1, 2 or 3 amino acids. The allowed changes can be determined experimentally by systematically inserting or deleting amino acids in the protein using recombinant DNA technology and determining the activity of the resulting recombinant variants. This does not require more than routine experimentation for the skilled person.

[0040] "Analogs" may alternatively or additionally be defined as amino acid sequences that are similar to the amino acid sequences disclosed above and that comprise a highly conserved portion of the Mac-1 domain (SEQ ID NO: 5) or an amino acid sequence that is at least 95% homologous thereto. The inventors surprisingly found that this domain is primarily responsible for the unexpected immunogenic activity of the IdeSsuis protein and is sufficient in itself to provide immune protection to vaccinated animals. There are different serotypes of S. suis that partially show large variations in their amino acid sequences, thus resulting in homology levels as low as about 60%. However, the highly conserved Mac-1 domain shows only small variations between the different serotypes, for example 97.9% between serotype strains 2 and 7.

[0041] Therefore, it is accepted that the amino acids of the present invention (and nucleic acids encoding them) show higher levels of variation outside the Mac-1 domain than within the domain.

[0042] The term "fragment" may be defined in a similar manner (see above). It describes an amino acid sequence that is shorter than an analog (less than about 400 amino acids). It contains or consists of a highly conserved portion of the Mac-1 domain (SEQ ID NO: 5) or an amino acid sequence that is at least 95% homologous thereto. Optionally, a fragment may be defined as having IgM protease activity, although this is not an essential requirement. These fragments may be used as exclusive active ingredients of the vaccine according to the invention.

[0043] Therefore, in a preferred embodiment, the vaccine composition of the present invention comprises, consists essentially of, or consists of a protein comprising or consisting of the amino acid sequence of SEQ ID NO: 5 or an amino acid sequence at least 95% homologous thereto. In a more preferred embodiment, the protein (or a vector encoding the protein) is the only active or immunogenic component.

[0044] As used herein in the context of vaccine compositions, the term "comprising" means that further active or immunogenic components may be present. "Consisting of" means that no further components are present, and "consisting essentially of" means that certain further components may be present, i.e. those that do not substantially affect the basic characteristics of the vaccine (i.e. inactive or non-immunogenic ingredients).

[0045] In a preferred embodiment, the present invention provides a vaccine composition, which is essentially composed of an rIdeSsuis protein that is at least 60%, 70%, 85% or 95% homologous to the amino acid sequence of the protein IdeSsuis of SEQ ID NO: 1, and / or comprises or consists of the amino acid sequence of SEQ ID NO: 5 or an amino acid sequence that is at least 95% homologous thereto.

[0046] The above definition also applies mutatis mutandis to the nucleic acid sequences encoding these proteins of the present invention. The definition of homology is the same, and the fragment length will be less than about 1200 nucleic acids.

[0047] Further preferred according to the present invention is a vaccine composition, wherein an effective amount of a fragment of a polypeptide according to (a) or (b) is part of a fusion protein with at least one other protein.

[0048] Preferred according to the present invention is a vaccine composition wherein the polynucleotide of (c) comprises a sequence encoding a protein defined as IdeSsuis, i.e.

[0049] (aa) SEQ ID NO: 1 amino acid sequence;

[0050] (ab) a fragment or analog of the amino acid sequence of SEQ ID NO: 1; or

[0051] (ac) (aa) or (ab) fragments having IgM protease activity.

[0052] According to the present invention, a vaccine composition is particularly preferred, characterized in that the polynucleotide comprises

[0053] (ca) SEQ ID NO: 3 or its complementary sequence;

[0054] (cb)(ca) sequence fragment or

[0055] (cc) A (ca) sequence fragment encoding a protein having IgM protease activity.

[0056] Further preferred according to the present invention is a vaccine composition, wherein the polynucleotide of (d) comprises a sequence encoding a protein defined as rIdeSsuis, i.e.

[0057] (ba) amino acid sequence of SEQ ID NO: 2;

[0058] (bb) a fragment or analog of the amino acid sequence of SEQ ID NO: 2;

[0059] (bc) an amino acid sequence lacking the amino acids at positions 1 to 34 in the amino acid sequence of SEQ ID NO: 1;

[0060] (bd) an amino acid sequence having at least 60% homology, preferably 70% homology, most preferably 85% homology to the amino acid sequence of protein IdeSsuis of SEQ ID NO: 1; or

[0061] (be) a fragment of (ba) or (bb) or (bc) or (bd) having IgM protease activity; and / or

[0062] (bf) a fragment of (ba) or (bb) or (bc) or (bd) comprising or consisting of the amino acid sequence of SEQ ID NO: 5 or an amino acid sequence that is at least 95% homologous thereto.

[0063] Particularly preferred according to the present invention is a vaccine composition, characterized in that the polynucleotide comprises

[0064] (da) SEQ ID NO: 4, 8 or 9 or a complementary sequence thereof;

[0065] (db)(da) sequence fragment or

[0066] (dc) A fragment of the (da) sequence encoding a protein having IgM protease activity.

[0067] Preferred according to the present invention is a vaccine composition wherein the polynucleotide is cDNA, DNA or cRNA, RNA. The term "nucleic acid sequence" refers to a heteropolymer of nucleotides or a sequence of these nucleotides. The terms "nucleic acid" and "polynucleotide" are used interchangeably herein to refer to a heteropolymer of nucleotides.

[0068] Furthermore, a preferred vaccine composition according to the present invention is also characterized in that the polynucleotide is integrated into a vector, wherein the polynucleotide is operably linked to the expression control region of the vector.

[0069] The expression vector preferably comprises one or more regulatory sequences. The term "expression vector" generally refers to a plasmid or a bacteriophage or a virus or a vector for expressing a polypeptide from a DNA (RNA) sequence. An expression vector may comprise a transcription unit comprising components (1) a genetic element or element having a regulatory role in gene expression, such as a promoter or an enhancer, (2) a structural or coding sequence transcribed into mRNA and translated into a protein, and (3) suitable transcription initiation and termination sequences. The structural unit intended for use in a yeast or eukaryotic expression system preferably comprises a leader sequence that enables the translated protein to be secreted extracellularly by the host cell. Alternatively, when the recombinant protein is expressed without a leader or transport sequence, it may include an N-terminal methionine residue. This residue may or may not be subsequently cleaved from the expressed recombinant protein to provide a final product.

[0070] It is also preferred that the vaccine composition according to the invention is provided in a physiologically administrable form and is suitable for intramuscular, intravenous, subcutaneous or cutaneous injection or mucosal administration. It should be noted that intravenous administration is less preferred.

[0071] In another aspect, the present invention relates to a fragment of IdeSsuis having the amino acid sequence of SEQ ID NO: 5, or an amino acid sequence at least 95% homologous thereto. SEQ ID NO: 5 corresponds to a highly conserved portion of the Mac-1 domain. Although this domain exhibits IgM protease activity, the immunogenic effect is not necessarily associated with this activity. For example, within the scope of the present invention are analogs of SEQ ID NO: 5 in which the active center of the protease has been inactivated by mutagenesis of the Cys residues. Also in this case, the analogs are effective as vaccines that elicit an immune response.

[0072] Furthermore, it was demonstrated that an amino acid sequence can be effective as a vaccine against S. suis infection if it retains at least 95% homology to SEQ ID NO: 5. This includes substitutions, insertions or deletions of single amino acids. It is demonstrated that naturally occurring Mac-1 domains, although showing some variations, do not differ by more than 5%, or in other words, have 95% or more identity in this domain. Exemplary S. suis sequences were obtained from strains isolated from different regions (Americas, Asia, Europe) and from different host organisms (humans, pigs). These strains belong to different serotypes (1 to 4, 7 to 9, 14 and 16 or non-typed). This is summarized in Appendix 1:

[0073]

[0074]

[0075] Sequence information about the Mac-1 domain of proteins WP_044671938, WP_002935529, WP_015647040, WP_023370787 and WP_044678723 is disclosed in SEQ ID NO: 10 to SEQ ID NO: 14. As their homology to SEQ ID NO: 5 is higher than 95%, they fall within the definition of fragments or homologs of the present invention.

[0076] Another aspect is the rIdeSsuis protein comprising the amino acid sequence of SEQ ID NO: 6 or 7, or an amino acid sequence that is at least 60%, preferably 70%, 85% or 95% homologous to the amino acid sequence of the protein IdeSsuis of SEQ ID NO: 6 or 7.

[0077] Another object of the present invention is to transfect a host cell with the vector.

[0078] Another object of the invention is a method for producing a protein defined as rIdeSsuis, as a guest antigen in a vector or in a different organism, the host cells being respectively transfected under conditions suitable for the expression of said recombinant protein.

[0079] Another aspect of the invention is an antibody recognizing the IdeSsuis or rIdeSsuis protein, analog or fragment as defined above.

[0080] The antibody is preferably selected from the group consisting of polyclonal antibodies, monoclonal antibodies, humanized antibodies, chimeric antibodies and synthetic antibodies.

[0081] The term "antibody" is used herein for complete antibodies and antibody fragments having a certain ability to selectively bind epitopes. Such fragments include, but are not limited to, Fab, F(ab')2, and Fv antibody fragments. The term "epitope" refers to any antigenic determinant of an antigen to which the paratope of an antibody can bind. Epitope determinants are usually composed of chemically active surface groups of molecules (e.g., amino acids or sugar residues) and usually display a three-dimensional structure as well as specific physical properties.

[0082] The antibodies according to the present invention can be produced according to any known procedure. For example, the pure whole IdeSsuis or rIdeSsuis protein or its fragment / analog according to the present invention can be produced and used as an immunogen to immunize animals and produce specific antibodies.

[0083] The production of polyclonal antibodies is well known. Detailed protocols can be found in, for example, Production of Polyclonal Antisera, in Immunochemical Protocols (Manson, editor), pages 1-5 (Humana Press 1992), published by Green et al., and Production of Polyclonal Antisera in Rabbits, Rats, Mice and Hamsters, in Current Protocols In Immunology, section 2.4.1 (1992), published by Coligan et al., Current Protocols in Immunology, section 2.4.1 (1992). Furthermore, the expert is familiar with several techniques for the purification and concentration of polyclonal as well as monoclonal antibodies (Coligan et al., Unit 9, Current Protocols in Immunology, Wiley Interscience, 1994).

[0084] The production of monoclonal antibodies is well known. Examples include the hybridoma method (Kohler and Milstein, 1975, Nature, 256:495-497, Coligan et al., Sections 2.5.1-2.6.7; and Harlow et al., Antibodies: A Laboratory Manual, p. 726 (Cold Spring Harbor Press, 1988)), trioma technology, human B cell hybridoma technology (Kozbor et al., 1983, Immunity Today, 4:72) and EBV hybridoma technology to produce human monoclonal antibodies (Cole et al., 1985, Monoclonal Antibodies and Cancer Therapy, Alan R. Liss, Inc., pp. 77-96).

[0085] In short, monoclonal antibodies can be obtained by injecting a mixture containing a protein according to the present invention into mice. Antibodies produced in mice are examined by serum probes. In the case of sufficient antibody titers, mice are killed and spleens are removed to separate B cells. B cells are fused with myeloma cells to obtain hybridomas. Hybridomas are cloned and clones are analyzed. Positive clones containing monoclonal antibodies against the protein are selected and antibodies are isolated from hybridoma cultures. There are many mature techniques to separate and purify monoclonal antibodies. Such techniques include affinity chromatography, size exclusion chromatography, and ion exchange chromatography using protein A agarose. See also, e.g., Coligan et al., section 2.7.1-2.7.12 and section “Immunglobulin G (IgG)”, in Methods In Molecular Biology, volume 10, pages 79-104 (Humana Press 1992).

[0086] Preferably, the present invention provides humanized IdeSsuis or rIdeSsuis-specific mouse antibodies.

[0087] The above antibodies may form part of a parenteral composition for therapeutic treatment of a human or animal (pig) patient suffering from an S. suis infection. However, it may also be used for prophylactic purposes.

[0088] In another aspect, the present invention relates to the use of a protein as disclosed above for the production of an antibody as described above.

[0089] Another object of the invention is the use of a vaccine or parenteral composition according to the invention to prevent or metaphylactic or therapeutic treatment of Streptococcus suis infection in pigs. It is further contemplated herein to use a vaccine or parenteral composition according to the invention for preventing or metaphylactic or therapeutic treatment of Streptococcus suis infection in human patients.

[0090] Particularly preferred according to the invention is the use of a vaccine composition, wherein said treatment induces an immune response in pigs, which immune response is the activation of humoral and cellular responses against the protein IdeSsuis produced by S. suis.

[0091] Treatment (vaccination) includes at least one or two immunizations. The total dose administered per pig / human is about 0.05-2.0 mg protein.

[0092] The preparation of vaccine compositions according to the present invention is known in the art and is described in manuals known to those skilled in the art. In order to produce vaccine compositions according to the present invention, pharmaceutically acceptable carriers, diluents or adjuvants can be used, including but not limited to the following: mineral salt adjuvants (e.g., aluminum-, calcium-, iron-, zirconium-based), tension active adjuvants (e.g., Quil A, QS-21, other saponins), bacteria-derived adjuvants (e.g., N-acetylmuramoyl-L-alanyl-D-isoglutamine (MDP), lipopolysaccharide (LPS), monophosphoryl lipid A, trehalose dimycolate (TDM), DNA, CpGs, bacterial toxins), adjuvant emulsions (e.g., FIA, Montanide, Adjuvant 65, Lipovant), liposome adjuvants, polymeric adjuvants and carriers, cytokines (e.g., granulocyte-macrophage colony stimulating factor), carbohydrate adjuvants, live antigen delivery systems (e.g., bacteria, viruses). In addition, carriers can also include dry preparations, such as coated patches made of titanium or polymers. Techniques for the preparation and administration of the vaccines of the present application can also be found in "Remington, The Science and Practice of Pharmacy", 22nd Edition.

[0093] Therefore, the present invention relates to a vaccine composition comprising a protein called IdeSsuis or rIdeSsuis or a fragment of either thereof; or a polynucleotide expressing the protein IdeSsuis or rIdeSsuis or a fragment of either thereof, which is integrated into an expression vector, and a recombinant protein is preferred.

[0094] The inventors detected the following:

[0095] (a) The induction of opsonized antibodies is crucial for the protective efficacy of S. suis vaccines.

[0096] (b) IdeSsuis promotes the survival of S. suis in the blood of vaccinated piglets.

[0097] Furthermore, the inventors have shown that vaccination of pigs using the individual protein rIdeSsuis as the sole antigen provides protection against pigs infected with S. suis. According to the invention, vaccination with rIdeSsuis prevents the cleavage of IgM by the IdeSsuis IgM protease of S. suis by inducing neutralizing antibodies.

[0098] According to the present invention, vaccination of pigs with the recombinant protein rIdeSsuis or a fragment thereof results in high titers of IdeSsuis-specific IgG antibodies with neutralizing activity compared to piglets immunized with convalescent or S. suis bacterin. In addition, according to Example 1, it has been shown that rIdeSsuis provides higher immunity against S. suis than bacterin vaccines of the prior art.

[0099] Furthermore, the inventors have shown that vaccination of pigs with the recombinant protein rIdeSsuis or an analog or fragment thereof reduces the survival of S. suis in the blood.

[0100] As mentioned above, an important aspect of the present invention lies in the surprising insight that the proteins, nucleic acids and their analogs and fragments as defined above can be used to provide protection against S. suis infection. only Immunogenic agents. No other active ingredients are required, such as the bacteria used in the prior art. This is supported by the experimental evidence provided in Example 2. It is shown that the vaccine according to the present invention has significantly enhanced efficacy relative to the placebo / control vaccine, see the bactericidal assay data of Experiments 1 and 2 of Example 2 ( Figure 3 and 4 ). The following conclusions can be drawn from the experimental results:

[0101] rIdeSsuis (SEQ ID NO: 2) provides immune protection across different serotypes of Streptococcus suis,

[0102] A protein containing at least the highly conserved Mac-1 domain (SEQ ID NO: 5) is sufficient to provide immune protection,

[0103] • In addition, IdeSsuis proteins of other serotypes containing at least the highly conserved Mac-1 domain (SEQ ID NO: 5) induced protection even though their overall sequence outside this domain differed from that of serotype 2 strains (even though some sequence segments were completely missing).

[0104] According to the present invention, the protein IdeSsuis or rIdeSsuis or its analogs / fragments can also be used for fusion proteins. Fusion proteins are produced by connecting two or more genes that originally encode separate proteins. Translation of the fusion gene results in a single or multiple polypeptides with each function derived from the original protein. In the prior art, fusion proteins are often used to simplify specific applications, such as detection, integration or transport of target proteins. A prominent member detected by fluorescence microscopy is the green fluorescent protein (GFP) fused to the target protein.

[0105] Other proteins that can be fused to IdeSsuis to improve antigen delivery and immunogenicity are immunoglobulin Fc fragments, non-toxic cholera toxin CTA subunits, mutant heat-labile toxins, Bacillus subtilis spore coat proteins, or bacterial flagellar proteins. In addition, IdeSsuis can be expressed on the surface of viral particles or virus-like particles using proteins fused to proteins of viruses or phages (e.g., modified vaccinia virus Ankara (MVA), hepatitis B virus, lambda phage, or filamentous phages such as fd, M13, or fI).

[0106] Another possibility for detecting fusion proteins is the use of so-called protein tags, which are often used during the production of fusion proteins for purification and detection by affinity chromatography, immunoblotting, immunohistochemistry or fluorescence microscopy, respectively. Protein tags are usually short amino acid sequences, such as HIS-tag, myc-tag, HA-tag, Step-tag, GST-tag, maltose binding protein tag or thioredoxin tag.

[0107] Methods for producing recombinant proteins such as rIdeSsuis, fragments thereof or analogs according to the present invention are well known to those skilled in the art and are also described by handbooks known to those skilled in the art. Typically, host cells are transfected with vectors encoding the target protein to produce the recombinant protein. Typically, those host cells can be bacteria (e.g., Escherichia coli, Bacillus or Lactococcus strains), people (e.g., 293-T, HEK-293), mouse cell lines, insect cell lines, yeast cells or plant-based systems.

[0108] For transfection of host cells, expression vectors such as plasmids (eg, pET, pQE), viruses and phages (eg, baculovirus, lambda phage or filamentous phage) can be used.

[0109] Typical vaccines or parenteral compositions are prepared as injections, either as liquid solutions or suspensions.

[0110] A subject of the invention is also a vaccine or a parenteral composition for subcutaneous, intravenous, intramuscular, cutaneous or mucosal application.

[0111] Vaccine of the present invention is used for the preventive or intermittent or therapeutic treatment of Streptococcus suis infection in pigs or humans. Treatment includes at least one, preferably two immunizations. Although a single immunization is preferred in practice, standard immunization generally includes a primary immunization-boosting scheme, i.e. 2 different inoculations. Booster inoculations are generally given within a time range of 1-3 weeks, preferably about 2 weeks, after the initial immunization. The dosage of a single inoculation may be identical or different, but preferably the vaccine dosage is identical for two times.

[0112] The total dose that must be administered to an animal or human patient is about 0.05-2.0 mg of the IdeSsuis or rIdeSsuis protein, analog or fragment as defined above. Preferred doses include 0.1-1.0, more preferably about 0.5 mg. If a single vaccination is sufficient, the dose is administered in one dose. If more than one vaccination is applied, the total dose is divided into several equal sub-doses, for example, if two vaccinations are used, the single doses of the vaccination are about 0.025-1.0 mg of protein. BRIEF DESCRIPTION OF THE DRAWINGS

[0113] The present invention is further described with reference to the following drawings, in which

[0114] Figure 1 A timeline representing a S. suis inoculation challenge experiment in pigs is shown,

[0115] Figure 2 Shown Figure 1 Figure 2 shows the results obtained from the inoculation challenge experiment.

[0116] Figure 3 A graph of a bactericidal assay including inoculation with placebo versus inoculation with rIdeSsuis is shown, and

[0117] Figure 4 Depicted are graphs of bactericidal assays involving inoculation with rIdeSsuis or rIdeSsuis analogs relative to a control group.

[0118] In addition, a sequence listing is attached to this specification.

[0119] SEQ ID NO: 1 shows the amino acid sequence of protein IdeSsuis;

[0120] SEQ ID NO: 2 shows the amino acid sequence of the recombinant protein rIdeSsuis without a signal peptide;

[0121] SEQ ID NO: 3 shows the nucleotide sequence encoding IdeSsuis and

[0122] SEQ ID NO: 4 shows the nucleotide sequence encoding rIdeSsuis.

[0123] SEQ ID NO: 5 shows the amino acid sequence of a highly conserved portion of the Mac-1 domain of IdeSsuis.

[0124] SEQ ID NO: 6 shows the amino acid sequence of rIdeSsuis_homolog, an analog of rIdeSsuis.

[0125] SEQ ID NO: 7 shows the amino acid sequence of the rIdeSsuis analog rIdeSsuisB2.

[0126] SEQ ID NO: 8 shows the nucleotide sequence encoding the rIdeSsuis analog rIdeSsuis_homolog.

[0127] SEQ ID NO: 9 shows the nucleotide sequence encoding the rIdeSsuis analog rIdeSsuisB2.

[0128] SEQ ID NO: 10 shows the amino acid sequence of amino acids 91 to 425 of WP_044671938.

[0129] SEQ ID NO: 11 shows the amino acid sequence of amino acids 91 to 425 of WP_002935529.

[0130] SEQ ID NO: 12 shows the amino acid sequence of amino acids 92 to 426 of WP_015647040.

[0131] SEQ ID NO: 13 shows the amino acid sequence of amino acids 92 to 426 of WP_023370787.

[0132] SEQ ID NO: 14 shows the amino acid sequence of amino acids 92 to 426 of WP_044678723. DETAILED DESCRIPTION

[0133] The following examples will explain the present invention. These examples should be understood as only a preferred embodiment of the present invention. This does not mean that the present invention is limited to the scope of the given examples.

[0134] Example 1

[0135] The following examples provide experimental data following a vaccination challenge experiment in piglets infected with S. suis.

[0136] An inoculation challenge experiment was set up in piglets with different S. suis serotypes (serotypes 2 and 9) and infection routes (intravenous and intranasal).

[0137] Briefly as Figure 1 As shown, 5-week-old piglets were primed with the rIdeSsuis vaccine. At 7 weeks of age, these piglets were boosted with rIdeSsuis and in one group were also primed with the bacterin (at Figure 1). One group of piglets was primed with the bacterin only at 7 weeks of age, and the last group of animals was vaccinated twice with a placebo consisting of buffer and adjuvant. Two weeks after the second immunization, the piglets were challenged intranasally. The animals were further monitored every 8 hours. For reasons of animal welfare, in any case in which the piglets showed a combination of high fever with apathy and anorexia, as well as in the case of clinical signs of acute polyarthritis or severe meningitis, the animals were euthanized.

[0138] All surviving piglets were sacrificed 14 days after infection.

[0139] Experiments have shown that immunization of piglets with the recombinant protein rIdeSsuis itself is sufficient to protect piglets from infection with S. suis serotype 2 ( Figure 2 ).

[0140] Example 2

[0141] In this embodiment, bactericidal assays have been used to evaluate the effectiveness of a given vaccine. This test involves determining its survival after a certain serotype of Streptococcus suis bacteria is added to the blood of a test animal. If there are protective antibodies for a certain serotype in the blood of the test animal, then the bacteria will be effectively killed within a 2-hour incubation time. The degree of protection is referred to as "survival factor" (SF), and is the ratio of the colony count after bacteria are added to the blood of the test animal for 120 minutes to the colony count directly after bacteria are added to the blood of the test animal. Low survival factor means efficient killing of bacteria in the blood, and therefore effectively protects the test animal.

[0142] Expression and purification of recombinant IdeSsuis (SEQ ID NO: 2), recombinant IdeSsuisB2 (SEQ ID NO: 7) and recombinant IdeSsuis_homolog (SEQ ID NO: 6) were performed by growing the appropriate strains in LB broth plus ampicillin. Protein expression was induced by adding IPTG. The expression of proteins was carried out under natural conditions by Ni as recommended by the manufacturer (Macherey-Nagel). 2+ The recombinant protein was purified by nitrilotriacetic acid affinity chromatography.

[0143] Immunization of piglets: Piglets were primed and boosted with 1.5 ml of vaccine containing 0.25 mg rIdeSsuis or 0.25 mg rIdeSsuisB2 or 0.5 mg rIdeSsuis_homolog (containing 20% ​​[vol / vol] Emulsigen as adjuvant).

[0144] Test 1:

[0145] Two or four litters were randomly divided into two experimental groups (n=9 / group), group 1 was control (placebo), and group 2 was immunized with rIdeSsuis (SEQ ID NO: 2). Animals were immunized and tested according to the following test protocol:

[0146]

[0147]

[0148] Figure 3 The results achieved are shown.

[0149] The control group (placebo) showed much higher survival factor than the vaccinated group. The recombinant antigen rIdeSsuis (vaccinated group) (SEQ ID NO: 2) containing the complete sequence of IdeSsuis protein of serotype 2 strain induced the production of antibodies that efficiently killed strain 2 as well as strain 9 S. suis bacteria.

[0150] Test 2:

[0151] Four littermates were randomly assigned to four experimental groups (n=6 / group, except for group 4, where n=5), group 1 control (unimmunized), group 2 immunized with rIdeSsuis_homolog (SEQ ID NO: 6), group 3 immunized with IdeSsuis derived from serotype 7 strain (rIdeSsuisB2; SEQ ID NO: 7), and group 4 immunized with SEQ ID NO: 2. Animals were immunized and tested according to the following test protocol:

[0152]

[0153]

[0154] Figure 4 The results achieved are shown.

[0155] The results show that the survival factor in the control group (group 1) is significantly higher than in the three vaccinated groups. The recombinant antigen rIdeSsuis_homolog (group 2), which contains only the N-terminal fragment containing the highly conserved Mac-1 domain (IgM-protease-domain), induced antibodies that led to a much better bacterial killing than in the control group. The results of group 2 can be compared with those obtained in group 4. The antigen rIdeSsuis, which contains the complete amino acid sequence of the mature IdeSsuis protein of the S. suis serotype 2 strain, also induced antibodies that significantly reduced the survival of the S. suis serotype 9 strain in the blood. The survival of the serotype 9 strain was even further impaired by the antibodies already induced by the antigen rIdeSsuisB2 (group 3).

[0156] rIdeSsuisB2 (SEQ ID NO: 7) contains the complete amino acid sequence of the mature IdeSsuis protein of the S. suis serotype 7 strain and differs in the C-terminal half of the protein as it lacks 114 aa of sequence compared to SEQ ID NO: 1. Aa 80 to 414 of SEQ ID NO: 7 (the so-called highly conserved part of the Mac-1 domain) corresponds to 97.9% of the sequence of SEQ ID NO: 5. The identity between the remaining C-terminal part of SEQ ID NO: 7 and SEQ ID NO: 1 is 96.4%.

[0157] The following conclusions can be drawn from the experimental results:

[0158] rIdeSsuis (SEQ ID NO: 2) provides immune protection across different serotypes of S. suis,

[0159] The amino acids containing at least the highly conserved Mac-1 domain are sufficient to provide immune protection,

[0160] • Furthermore, IdeSsuis proteins of other serotypes containing at least the highly conserved Mac-1 domain induced protection even though their overall sequence outside the domain differed from that of serotype 2 strains (even though some sequence segments were completely missing). Sequence Listing <110> Chua Animal Health <120> Vaccine composition against Streptococcus suis infection <130> P37819-WO <150> EP14170637.4 <151> 2014-05-30 <160> 14 <170> PatentIn version 3.5 <210> 1 <211> 1141 <212> PRT <213> Streptococcus suis <400> 1 Met Asn Ile Gln Glu Arg Phe Ser Leu Arg Lys Ser Ala Val Gly Leu 1 5 10 15 Val Ser Val Ser Leu Leu Cys Ala Ile Tyr Thr Ser Thr Val Ala Ala 20 25 30 Asp Thr Val Val Thr Gly Val Asn Glu Ile Ile Glu Glu Ser Gln Val 35 40 45 Lys Asp Glu Val Ser Ile Glu Ser Glu Lys Asn Glu Ser Leu Asp Gly 50 55 60 Ser Asn Ile Glu Ile Val Glu Glu Ile Ala Asp Asn Ile Pro Ser Pro 65 70 75 80 Val Ile Ala Glu Gly Glu Val Ala Val Glu Met Lys Val Asp Arg Gly 85 90 95 Thr Glu Asn Val Val Ser Arg Asn Asp Thr Glu Val Thr Thr Ser Glu 100 105 110 Gln Asn Gln Ile Glu Val Thr Glu Thr Lys Glu Ile Leu Asn Gln Thr 115 120 125 Ser Tyr Gln Thr Glu Ser Gly Glu Gln Arg Gln Ile Ile Trp Ala His 130 135 140 Gly Ile Thr Pro Pro Ala Met Glu Gln Ser Gly Gly Phe Val Lys Glu 145 150 155 160 Lys Tyr Gly Asp Tyr Leu Asn Tyr Thr Ala Pro Phe Glu Ala Gly Lys 165 170 175 Gly Tyr Tyr Asp Thr Asn Lys Ser Leu Asn Ala Ser Phe Ile Asp Leu 180 185 190 Asn Leu Cys Phe Ala Ala Val Ser Ser Asn Met Val His Trp Trp Leu 195 200 205 Glu Gln Asn Ser Ser Tyr Val Glu Arg Tyr Leu Lys Glu Lys Lys Gly 210 215 220 Thr Val Asn Val Glu Glu Asn Tyr Ala Ile Thr Asp Leu Arg Arg Tyr 225 230 235 240 Ile Asn Ser Phe Gln Asn Gln Gln Asn Ser Arg Val Phe Asp Met Phe 245 250 255 Lys Thr Tyr Tyr Gly Tyr Arg Thr Asn Gly Phe Val Ser Asp Ala Leu 260 265 270 Val Asp Leu Phe Ile Asn Gly Tyr Lys Pro Lys Ala Gln Gly Gly Val 275 280 285 Asn Leu Glu Asp Ser Gln Leu Val Pro Asp Ser Arg Gly Gly Phe Phe 290 295 300 Tyr Asp Val Phe Lys Glu Lys Lys Leu Thr Asn Arg Ile Phe Ser Gly 305 310 315 320 Ser Tyr Glu Arg Phe Gly Glu Asp Val Arg Thr Val Leu Glu Ser Lys 325 330 335 Gly Leu Leu Gly Leu Thr Tyr Arg Thr Leu Gly Tyr Ala Thr His Ile 340 345 350 Val Thr Val Trp Gly Ala Glu Tyr Asp Asn Gln Gly Lys Ile Lys Ala 355 360 365 Val Tyr Ile Thr Asp Ser Asp Asp Gln Gln Glu Gln Ile Gly Leu Lys 370 375 380 Arg Met Gly Ile Thr Arg Asp Ala Ser Gly Asn Pro Arg Leu Asn Asn 385 390 395 400 His Met Lys Asn Asn Ser Ala Gly Ala Leu Leu Asp Tyr Val His Thr 405 410 415 Ile Arg Leu Gly Gln Asp Leu Trp Glu Glu Tyr Phe Asn Pro Leu Ala 420 425 430 Lys Ala Lys Glu Thr Ala Ser Gln Thr Leu Ala Asp Thr Lys Lys Ala 435 440 445 Leu Asp Leu Ser Ile Gln Gly Gln Ser Glu Leu Pro Glu Ser Met Arg 450 455 460 Leu Ile Tyr Leu Glu Lys Leu Asn Asn Leu Tyr Asn Gln Gly Ile Leu 465 470 475 480 Ser Ile Gln Lys Ala Glu Ser Ser Glu Met Leu Ser Gly Ala Leu Glu 485 490 495 Asn Gly Leu Asn Ser Leu Lys Ser Leu Asp Phe Pro Ile Ser Glu Val 500 505 510 Gly Asn Ala Leu Ala Pro Asp Leu Pro Val Gly Asp Arg Ser Thr Val 515 520 525 Ser Asp Val Asp Ser Leu Ser Ser Gln Glu Thr Ser Ser Thr Asn Leu 530 535 540 Glu Ala Asp Thr Glu Asn Ala Gly Ile Ile Ala Asp Gly Thr Asn Gln 545 550 555 560 Leu His Phe Pro Val Glu Ala Gln Thr Thr Ser Ser Val Glu Ala Glu 565 570 575 Gly Asp Asn Val Phe Glu Gln Glu Ala Asp Thr Leu Pro Ile Ile Ile 580 585 590 Glu Asn Lys Asp Glu Phe Gly Ser Glu Leu Ser Arg Asn Met Gln Thr 595 600 605 Ser Glu Thr Asp Ser Leu Val Val Ala Val Glu Glu Asp Val Lys Asn 610 615 620 Asp Glu Val Ala Gln Val Glu Glu Leu Leu Glu Ser Glu Lys Val Glu 625 630 635 640 Asn Gln Ser Ser Glu Leu Leu Ser Asp Thr Leu Ile Val Glu Ser Ala 645 650 655 Asn Asp Lys Glu Glu Asp Arg Val Glu Ala Val Val Ser Glu Gln Pro 660 665 670 Asp Ser Ile Pro His Gln Asn Val Glu Ile Ser Leu Val Glu Pro Thr 675 680 685 Asn Val Glu Thr Glu Thr Val Val Thr Pro Ile Asn Asp Ala Ala Thr 690 695 700 Pro His Gly Ser Pro Thr Tyr Ile Asp Asn Ser Val Thr Glu Ser Val 705 710 715 720 Ala Thr Pro Leu Glu Lys Asp Ser Ile Gln Ala Gly Glu Thr Glu Ile 725 730 735 Ala Glu Pro Thr Ser Ser Glu Ser Thr Asn Val Glu Thr Glu Thr Val 740 745 750 Val Thr Pro Val Asn Asp Val Ala Thr Pro His Gly Ser Pro Thr Tyr 755 760 765 Ile Asp Asn Ser Val Thr Glu Ser Val Ala Thr Pro Leu Glu Lys Asp 770 775 780 Ser Ile Gln Ala Gly Glu Thr Glu Ile Ala Glu Pro Thr Ser Ser Glu 785 790 795 800 Ser Thr Asn Val Glu Thr Glu Thr Val Val Thr Pro Val Asn Asp Val 805 810 815 Ala Thr Pro His Gly Ser Pro Thr Tyr Ile Asp Asn Ser Val Thr Glu 820 825 830 Ser Val Ala Thr Pro Leu Glu Lys Asp Ser Ile Gln Ala Gly Glu Thr 835 840 845 Glu Ile Ala Glu Pro Thr Ser Ser Glu Ser Thr Ser Val Glu Ala Glu 850 855 860 Leu Val Asp Asn Ser Glu Ile His Ala Ala Thr Ser Ser Val Thr Pro 865 870 875 880 Cys Gly Ser Ser Ala Tyr Ala Asp Gly Ser Thr Thr Glu Ser Val Ala 885 890 895 Thr Pro Leu Glu Lys Asp Ser Ile Gln Thr Gly Asn Thr Glu Ile Ala 900 905 910 Glu Pro Thr Ser Ser Lys Ser Thr Asn Val Glu Ala Ala Ser Val Asp 915 920 925 Asn Ser Glu Ile His Ala Asp Ala Ser Leu Thr Ala Val Ser Ser Val 930 935 940 Asn Leu Asp Asn Pro Val Ile Glu Pro Val Ala Ile Ser Leu Ile Gly 945 950 955 960 Ser Lys Arg Asp Thr Asn Ala Glu Val Glu Val Ser Ser Leu Ser Lys 965 970 975 Arg Glu Val Arg Lys Thr Asn Thr Asp Gly Leu Ile Ser Val Gln Ser 980 985 990 Lys Val Ile Lys Lys Glu Leu Leu Glu Ser Ser Leu Ala Glu Ala Gly 995 1000 1005 Ser Pro Leu Leu Glu Ala Thr Ile Ala Gln Ser Ser Asn Ser Asn 1010 1015 1020 Ser Thr Glu Ile Gly Met Ser Tyr Gln Asn Thr Val Leu Leu Glu 1025 1030 1035 Ser Asn Asn Thr Glu Arg Gln Val Ser Lys Ala Glu Ile Val Met 1040 1045 1050 Glu His Lys Glu Thr Glu Leu Val Glu Thr Val Ser Ser Ala Ser 1055 1060 1065 Glu Pro Val Val Leu Val Glu Asn Ile Ser Gln Thr Ser Asn Asn 1070 1075 1080 Thr Ile Glu Ser Gly Lys Asn Met Gly Val Gln Ser Gln Ala Gly 1085 1090 1095 Ala Lys Gln Ile Leu Gly Val Glu Gln Ser Ser Lys Val Ser Thr 1100 1105 1110 Pro Thr Ser Arg Gln Ile Met Gly Val Gly Leu Leu Thr Leu Val 1115 1120 1125 Leu Gly Ser Ala Leu Gly Leu Leu Lys Lys Arg Arg Lys 1130 1135 1140 <210> 2 <211> 1130 <212> PRT <213> Artificial <220> <223> rIdeSsuis <400> 2 Met Ala His His His His His His Val Gly Thr Gly Ser Asn Asp Asp 1 5 10 15 Asp Asp Lys Ser Pro Asp Pro Val Val Thr Gly Val Asn Glu Ile Ile 20 25 30 Glu Glu Ser Gln Val Lys Asp Glu Val Ser Ile Glu Ser Glu Lys Asn 35 40 45 Glu Ser Leu Asp Gly Ser Asn Ile Glu Ile Val Glu Glu Ile Ala Asp 50 55 60 Asn Ile Pro Ser Pro Val Ile Ala Glu Gly Glu Val Ala Val Glu Met 65 70 75 80 Lys Val Asp Arg Gly Thr Glu Asn Val Val Ser Arg Asn Asp Thr Glu 85 90 95 Val Thr Thr Ser Glu Gln Asn Gln Ile Glu Val Thr Glu Thr Lys Glu 100 105 110 Ile Leu Asn Gln Thr Ser Tyr Gln Thr Glu Ser Gly Glu Gln Arg Gln 115 120 125 Ile Ile Trp Ala His Gly Ile Thr Pro Pro Ala Met Glu Gln Ser Gly 130 135 140 Gly Phe Val Lys Glu Lys Tyr Gly Asp Tyr Leu Asn Tyr Thr Ala Pro 145 150 155 160 Phe Glu Ala Gly Lys Gly Tyr Tyr Asp Thr Asn Lys Ser Leu Asn Ala 165 170 175 Ser Phe Ile Asp Leu Asn Leu Cys Phe Ala Ala Val Ser Ser Asn Met 180 185 190 Val His Trp Trp Leu Glu Gln Asn Ser Ser Tyr Val Glu Arg Tyr Leu 195 200 205 Lys Glu Lys Lys Gly Thr Val Asn Val Glu Glu Asn Tyr Ala Ile Thr 210 215 220 Asp Leu Arg Arg Tyr Ile Asn Ser Phe Gln Asn Gln Gln Asn Ser Arg 225 230 235 240 Val Phe Asp Met Phe Lys Thr Tyr Tyr Gly Tyr Arg Thr Asn Gly Phe 245 250 255 Val Ser Asp Ala Leu Val Asp Leu Phe Ile Asn Gly Tyr Lys Pro Lys 260 265 270 Ala Gln Gly Gly Val Asn Leu Glu Asp Ser Gln Leu Val Pro Asp Ser 275 280 285 Arg Gly Gly Phe Phe Tyr Asp Val Phe Lys Glu Lys Lys Leu Thr Asn 290 295 300 Arg Ile Phe Ser Gly Ser Tyr Glu Arg Phe Gly Glu Asp Val Arg Thr 305 310 315 320 Val Leu Glu Ser Lys Gly Leu Leu Gly Leu Thr Tyr Arg Thr Leu Gly 325 330 335 Tyr Ala Thr His Ile Val Thr Val Trp Gly Ala Glu Tyr Asp Asn Gln 340 345 350 Gly Lys Ile Lys Ala Val Tyr Ile Thr Asp Ser Asp Asp Gln Gln Glu 355 360 365 Gln Ile Gly Leu Lys Arg Met Gly Ile Thr Arg Asp Ala Ser Gly Asn 370 375 380 Pro Arg Leu Asn Asn His Met Lys Asn Asn Ser Ala Gly Ala Leu Leu 385 390 395 400 Asp Tyr Val His Thr Ile Arg Leu Gly Gln Asp Leu Trp Glu Glu Tyr 405 410 415 Phe Asn Pro Leu Ala Lys Ala Lys Glu Thr Ala Ser Gln Thr Leu Ala 420 425 430 Asp Thr Lys Lys Ala Leu Asp Leu Ser Ile Gln Gly Gln Ser Glu Leu 435 440 445 Pro Glu Ser Met Arg Leu Ile Tyr Leu Glu Lys Leu Asn Asn Leu Tyr 450 455 460 Asn Gln Gly Ile Leu Ser Ile Gln Lys Ala Glu Ser Ser Glu Met Leu 465 470 475 480 Ser Gly Ala Leu Glu Asn Gly Leu Asn Ser Leu Lys Ser Leu Asp Phe 485 490 495 Pro Ile Ser Glu Val Gly Asn Ala Leu Ala Pro Asp Leu Pro Val Gly 500 505 510 Asp Arg Ser Thr Val Ser Asp Val Asp Ser Leu Ser Ser Gln Glu Thr 515 520 525 Ser Ser Thr Asn Leu Glu Ala Asp Thr Glu Asn Ala Gly Ile Ile Ala 530 535 540 Asp Gly Thr Asn Gln Leu His Phe Pro Val Glu Ala Gln Thr Thr Ser 545 550 555 560 Ser Val Glu Ala Glu Gly Asp Asn Val Phe Glu Gln Glu Ala Asp Thr 565 570 575 Leu Pro Ile Ile Ile Glu Asn Lys Asp Glu Phe Gly Ser Glu Leu Ser 580 585 590 Arg Asn Met Gln Thr Ser Glu Thr Asp Ser Leu Val Val Ala Val Glu 595 600 605 Glu Asp Val Lys Asn Asp Glu Val Ala Gln Val Glu Glu Leu Leu Glu 610 615 620 Ser Glu Lys Val Glu Asn Gln Ser Ser Glu Leu Leu Ser Asp Thr Leu 625 630 635 640 Ile Val Glu Ser Ala Asn Asp Lys Glu Glu Asp Arg Val Glu Ala Val 645 650 655 Val Ser Glu Gln Pro Asp Ser Ile Pro His Gln Asn Val Glu Ile Ser 660 665 670 Leu Val Glu Pro Thr Asn Val Glu Thr Glu Thr Val Val Thr Pro Ile 675 680 685 Asn Asp Ala Ala Thr Pro His Gly Ser Pro Thr Tyr Ile Asp Asn Ser 690 695 700 Val Thr Glu Ser Val Ala Thr Pro Leu Glu Lys Asp Ser Ile Gln Ala 705 710 715 720 Gly Glu Thr Glu Ile Ala Glu Pro Thr Ser Ser Glu Ser Thr Asn Val 725 730 735 Glu Thr Glu Thr Val Val Thr Pro Val Asn Asp Val Ala Thr Pro His 740 745 750 Gly Ser Pro Thr Tyr Ile Asp Asn Ser Val Thr Glu Ser Val Ala Thr 755 760 765 Pro Leu Glu Lys Asp Ser Ile Gln Ala Gly Glu Thr Glu Ile Ala Glu 770 775 780 Pro Thr Ser Ser Glu Ser Thr Asn Val Glu Thr Glu Thr Val Val Thr 785 790 795 800 Pro Val Asn Asp Val Ala Thr Pro His Gly Ser Pro Thr Tyr Ile Asp 805 810 815 Asn Ser Val Thr Glu Ser Val Ala Thr Pro Leu Glu Lys Asp Ser Ile 820 825 830 Gln Ala Gly Glu Thr Glu Ile Ala Glu Pro Thr Ser Ser Glu Ser Thr 835 840 845 Ser Val Glu Ala Glu Leu Val Asp Asn Ser Glu Ile His Ala Ala Thr 850 855 860 Ser Ser Val Thr Pro Cys Gly Ser Ser Ala Tyr Ala Asp Gly Ser Thr 865 870 875 880 Thr Glu Ser Val Ala Thr Pro Leu Glu Lys Asp Ser Ile Gln Thr Gly 885 890 895 Asn Thr Glu Ile Ala Glu Pro Thr Ser Ser Lys Ser Thr Asn Val Glu 900 905 910 Ala Ala Ser Val Asp Asn Ser Glu Ile His Ala Asp Ala Ser Leu Thr 915 920 925 Ala Val Ser Ser Val Asn Leu Asp Asn Pro Val Ile Glu Pro Val Ala 930 935 940 Ile Ser Leu Ile Gly Ser Lys Arg Asp Thr Asn Ala Glu Val Glu Val 945 950 955 960 Ser Ser Leu Ser Lys Arg Glu Val Arg Lys Thr Asn Thr Asp Gly Leu 965 970 975 Ile Ser Val Gln Ser Lys Val Ile Lys Lys Glu Leu Leu Glu Ser Ser 980 985 990 Leu Ala Glu Ala Gly Ser Pro Leu Leu Glu Ala Thr Ile Ala Gln Ser 995 1000 1005 Ser Asn Ser Asn Ser Thr Glu Ile Gly Met Ser Tyr Gln Asn Thr 1010 1015 1020 Val Leu Leu Glu Ser Asn Asn Thr Glu Arg Gln Val Ser Lys Ala 1025 1030 1035 Glu Ile Val Met Glu His Lys Glu Thr Glu Leu Val Glu Thr Val 1040 1045 1050 Ser Ser Ala Ser Glu Pro Val Val Leu Val Glu Asn Ile Ser Gln 1055 1060 1065 Thr Ser Asn Asn Thr Ile Glu Ser Gly Lys Asn Met Gly Val Gln 1070 1075 1080 Ser Gln Ala Gly Ala Lys Gln Ile Leu Gly Val Glu Gln Ser Ser 1085 1090 1095 Lys Val Ser Thr Pro Thr Ser Arg Gln Ile Met Gly Val Gly Leu 1100 1105 1110 Leu Thr Leu Val Leu Gly Ser Ala Leu Gly Leu Leu Lys Lys Arg 1115 1120 1125 Arg Lys 1130 <210> 3 <211> 3426 <212> DNA <213> Streptococcus suis <400> 3 atgaacattc aagaacgatt ttctttgaga aaatccgcgg ttggcttggt ctcagtctct 60 ttgctatgtg ctatttatac atccactgtt gctgccgata cagttgttac aggagtgaat 120 gaaataattg aagaatcaca agtcaaggat gaggtatcta ttgaatcaga aaaaaatgaa 180 tccctagatg gttctaatat tgaaattgta gaggaaatag cagacaacat cccatcacct 240 gttatcgctg aagggaagt agcggtagag atgaagttg acagaggc cgagaatgta 300 gtttctagaa atgatacaga agttacgacg agcgagcaaa atcagataga ggttactgag 360 aaaaagaaa ttttgaatca gaccagttat haaacggaa gtggcgagca acgacaatt 420 atatgggccc atggaatttac tcctcctgca atggacaaa gcggtggtttt tgtaaggaa 480 aagtatggag actatttaaa ctatacagcg ccatttgagg ctggaaaagg ctactatgat 540 accaataaga gtctgaatgc ttcatttatt gaccttaatc ttgttttgc agctgtgtct 600 tcaaacatgg tacattggtg gttggaacag atagttcct atgttgagcg atatctcaa 660 gaaaaaaagg gtacagtaaa tgttgaagaa aactacgcaa taacggactt acggcgctat 720 atttaatcat tccaaatca acaaatagt cgagttttg atatgttca aacttactat 780 ggttatcgta caatggttt tgtatcagat gccttggttg acttgtttat taacggatat 840 aaacctaagg cacaggggcgg tgtcaatctg gaagatagcc agttagtacc agatagtagg 900 ggtggcttt tcacgacgt ttcaagg aaaaactga caatcgaat ttttagtggt 960 agttatgagc ggtttggtga ggatgttcga actgttttgg aaagcaaagg attackcggc ttaacttata gaacattagg ttatgcaacg catattgtga cggtatgggg tgctgagtac 1140. 1140. 1140. 1140. 1140. 1140. 1140. 1140. 1140. 1140. 1140 attggtttga agcgtatggg aatcactcgt gatgcttccg gaatccacg tttgaat catatgaaaa ataattcagc tggagcgctt ttggattatg tccatacaat ccgtctgggt caagacttat gggaagaata tttcaatccg cttgcaaaag ccaaagaaac agctagtcag acattagccg atacaaagaa ggcgttggat ttgtctattc aaggacaatc tgaattgcca 1440. 1440. 1440. 1440. 1440. 1440. 1440. 1440. 1440. 1440. 1440. 1440. 1440 tctattcaaa aggcagaaag ttctgagatg ctaagtggtg cattggaaaa tggtttaaat agtttaaaga gtttagattt tcctatttca aagttgga atgctttggc accagattta ccagtaggtg atcgctcaac ggtttcagat gttgattctc tatcatctca agaaacaagt tccacaaatt tggaagcaga cacagagaat gcaggtatta ttgcagatgg taccaatcaa ttgcattttc cagtggaggc ccaaacgaca tcttcagtag aggctgaggg agataatgtt tttgaacaag aggcagatac attaccaata attattgaaa acaaggatga atttggttca 1860. 1860. 1860. 1860. 1860. 1860. 1860. 1860. 1860. 1860 gatgtgaaaa atgatgaggt agcccaagtt gaagagcttc ttgaatcaga aaaagttgaa aatcagagtt cggaacttct gtcagacacc ctaatcgtag aggtgcaaa tgacaaagaa 2040. tggaggcggt tgtttctgaa caaccagact caataccaca tcaaaatgta gaatctctc ttgtagaacc aacgaatgtc gaatctgaa ctgtggtcac tcctattaat gatgcagcta ctcctcatgg ttccccgacg father attccgtac tgaatctgta gctactccac ttgaaaaaga ctccattcaa gccggggaga cagagattgc agaaccaacc tcgagcgaat caacgaatgt cgaaactgaa actgtggtca ctcctgttaa tgatgtagct actcctcatg gttccccgac gtatatagat aattccgtaa ctgaatctgt agctactcca cttgaaaaag actccattca agccggagag acagaattg cagaaccaac ctcgagcgaa tcaacgaatg tcgaaactga aactgtggtc actcctgtta atgatgtagc tactcctcat 2460 ggttccccga cgtatataga taattccgta actgaatctg tagctactcc acttgaaaaa 2520 gactccattc aagccgggga gacagagatt gcagaaccaa cctcgagcga atcaactagt 2580 gttgaagctg aacttgtcga caattctgaa attcatgcag ctacctcttc agttactccc 2640 tgtggctcct cggcatatgc agatggttcc acaactgaat ctgtagccac tccgcttgaa 2700 aaagactcca ttcagactgg aaatacagaa attgcagaac caacctcgag caaatcaact 2760 aatgtagaag ctgcatctgt cgacaattct gaaattcatg cagatgcctc tctaactgct 2820 gtttcatcag ttaatctgga taatccagtg attgaaccag tagctatctc ccttatcggt 2880 tctaagaggg acacgaatgc agaagtagaa gtttcttcat tatcgaaaag agaggttaga 2940 aaaacaaata ctgacgggct aatctctgtt caatcaaaag ttattaagaa agaattgcta 3000 gaatcaagct tagcagaagc agggtctcca ttgctagaag ccaccattgc tcagtcttca 3060 aactcaaata gtactgagat aggtatgagc tatcagaata ctgtgttatt agagtctaat 3120 aatacagagc gtcaggtgtc taaagcagaa attgttatgg aacacaagga gacagagtta 3180 gttgaaacgg tttcatctgc ttctgagcct gtagtgctcg tagaaaatat ctcacaaacc 3240 tcaaataata ctattgaatc tggtaagaat atgggagttc aatctcaagc aggtgcaaaa 3300 caaattttag gcgtagaaca atcttcgaaa gtaagtacac ctacttcaag acagattatg 3360 ggagtcggtc tattgactct tgttcttggt agtgctttag gtttgttaaa gaaaagacgt 3420 aagtaa 3426 <210> 4 <211> 3393 <212> DNA <213> Artificial <220> <223> Nucleotide sequence encoding rIdeSsuis <400> 4 atggcacatc accaccacca tcacgtgggt accggttcga atgatgacga cgacaagagt 60 ccggatccag ttgttacagg agtgaatgaa ataattgaag aatcacaagt caaggatgag 120 gtatctattg aatcagaaaa aaatgaatcc ctagatggtt ctaatattga aattgtagag 180 gaaatagcag acaacatccc atcacctgtt atcgctgaag gggaagtagc ggtagagatg 240 aaagttgaca gagggaccga gaatgtagtt tctagaaatg atacagaagt tacgacgagc 300 gagcaaatc agatagaggt tactgagaca aaagaaattt tgaatcagac cagttatca 360 acggagtg gcgagcaacg acaaattata tgggcccatg gatttactcc tcctgcaatg 420 gaacaaagcg gtggttttgt aaaggaaag tatggagact attackaacta tacagcgcca 480 tttgaggctg gaaaaggcta ctatgatacc addaagagtc tgaatgctc atttattgac 540 cttaatcttt gtttgcagc tgtgtctca aacatggtac attggtggtt ggaacagaat 600 agttcctatg ttgagcgata tctcaagaa aaaaagggta cagtaaatgt tgagaaaac 660 tacgcaataa cggacttacg gcgctatatt attcattcc aaatcaa aaatagtcga 720 gtttttgata tgttcaaac ttactatggt tatcgtacaa atggttttgt atcagatgcc 780 ttggttgact tgtttaa cggatataaa cctaggcac agggcggtgt caatctggaa 840 gatagccagt tagtaccaga tagtaggtt ggctttttct acgacgtttt caagagaaa 900 aaactgacaa atcgaatttt tagtggtagt tatgagcggt tggtgagga tgttcgaact 960 gttttggaaa gcaaggatt actcggctta acttatagaa cattaggtta tgcaacgcat 1020 attgtgacgg tatggggtgc tgagtacgac atcaggta agatttaggc tgtctatatc 1080 acagattctg atgatcaca agacaatt gttttgaagc gtatgggaat cactcgtgat 1140 gcttccggaa atccacgttt gataatcat atgaaaaata attcagctgg agcgctttg 1200 gattatgtcc atacaatccg tctgggtca gacttatggg agaatattt caatccgctt 1260 gcaaaagcca aagaaacagc tagtcagaca ttagccgata caaaaggc gttggattg 1320 tctattcaag gatactga attgccagaa tcaatgcgac tgatttact tgaaaaacta 1380 aaatctct atatcagg aattctatct attcaaagg cagaaagttc tgagatgcta 1440 agtggtgcat tggaaatgg ttaatagt ttaagagtt tagattttcc tatttcaga 1500 gttggaaatg ctttggcacc agatttacca gtaggtgatc gctcaacggt ttcagatgtt 1560 gattctctat catctcaaga aacaagttcc acaatttgg aagcagacac agagaatgca 1620 ggtattattg cagatggtac caatcattg cattttccag tggaggccca acgacatct 1680 tcagtagagg ctgagggaga taatgtttt gaacagagg cagatacatt accaatatt 1740 attgaaaaca aggatgaatt tggttcagaa ctatcagaa acatgcaac gtcagaaacg 1800 gattcgctag tagtagctt tgaagagat gtgaaaaatg atgaggtagc ccaagttgaa 1860 gagctcttg aatcagaaaa agttgaaat cagagttcgg aacttctgtc agacacccta 1920 atcgtagaga gtgcaatga CAagaaga gatgagtgg aggcggttgt ttctgaaca 1980 ccagactcaa taccacatca aaatgtagaa atctctcttg tagaccac gatgtcgaa 2040 actgaaactg tggtcactcc tattaatgat gcagctactc ctcatggttc cccgacgtat 2100 atagataatt ccgtactga atctgtagct actccacttg aaaagactc cattcaagcc 2160 ggggagacag agattgcaga accaactcg agcgaatcaa cgaatgtcga aactgaaact 2220 gtggtcactc ctgttaatga tgtagctact cctcatggtt ccccgacgta tatagatatat 2280 tccgtaactg aatctgtagc tactccactt gaaaagact ccattcaagc cggagagaca 2340 gaaattgcag aaccaacctc gagcgaatca acgaatgtcg aaactgaaac tgtggtcact 2400 cctgttaatg atgtagctac tcctcatggt tccccgacgt atatagataa ttccgtact 2460 gaatctgtag ctactccact tgaaaaagac tccattcaag ccggggagac agagattgca 2520 gaaccaacct cgagcgaatc aactagtgtt gaagctgaac ttgtcgacaa ttctgaaatt 2580 catgcagcta cctcttcagt tactccctgt ggctcctcgg catatgcaga tggttccaca 2640 actgaatctg tagccactcc gcttgaaaaa gactccattc agactggaaa tacagaaatt 2700 gcagaaccaa cctcgagcaa atcaactaat gtagaagctg catctgtcga caattctgaa 2760 attcatgcag atgcctctct aactgctgtt tcatcagtta atctggataa tccagtgatt 2820 gaaccagtag ctatctccct tatcggttct aagagggaca cgaatgcaga agtagaagtt 2880 tcttcattat cgaaaagaga ggttagaaaa acaaatactg acgggctaat ctctgttcaa 2940 tcaaaagtta ttaagaaaga attgctagaa tcaagcttag cagaagcagg gtctccattg 3000 ctagaagcca ccattgctca gtcttcaaac tcaaatagta ctgagatagg tatgagctat 3060 cagaatactg tgttattaga gtctaataat acagagcgtc aggtgtctaa agcagaaatt 3120 gttatggaac acaaggagac agagttagtt gaaacggttt catctgcttc tgagcctgta 3180 gtgctcgtag aaaatatctc acaaacctca aataatacta ttgaatctgg taagaatatg 3240 ggagttcaat ctcaagcagg tgcaaaacaa attttaggcg tagaacaatc ttcgaaagta 3300 agtacaccta cttcaagaca gattatggga gtcggtctat tgactcttgt tcttggtagt 3360 gctttaggtt tgttaaagaa aagacgtaag taa 3393 <210> 5 <211> 335 <212> PRT <213> Artificial <220> <223> Partial sequence of Mac-1 domain <400> 5 Met Lys Val Asp Arg Gly Thr Glu Asn Val Val Ser Arg Asn Asp Thr 1 5 10 15 Glu Val Thr Thr Ser Glu Gln Asn Gln Ile Glu Val Thr Glu Thr Lys 20 25 30 Glu Ile Leu Asn Gln Thr Ser Tyr Gln Thr Glu Ser Gly Glu Gln Arg 35 40 45 Gln Ile Ile Trp Ala His Gly Ile Thr Pro Pro Ala Met Glu Gln Ser 50 55 60 Gly Gly Phe Val Lys Glu Lys Tyr Gly Asp Tyr Leu Asn Tyr Thr Ala 65 70 75 80 Pro Phe Glu Ala Gly Lys Gly Tyr Tyr Asp Thr Asn Lys Ser Leu Asn 85 90 95 Ala Ser Phe Ile Asp Leu Asn Leu Cys Phe Ala Ala Val Ser Ser Asn 100 105 110 Met Val His Trp Trp Leu Glu Gln Asn Ser Ser Tyr Val Glu Arg Tyr 115 120 125 Leu Lys Glu Lys Lys Gly Thr Val Asn Val Glu Glu Asn Tyr Ala Ile 130 135 140 Thr Asp Leu Arg Arg Tyr Ile Asn Ser Phe Gln Asn Gln Gln Asn Ser 145 150 155 160 Arg Val Phe Asp Met Phe Lys Thr Tyr Tyr Gly Tyr Arg Thr Asn Gly 165 170 175 Phe Val Ser Asp Ala Leu Val Asp Leu Phe Ile Asn Gly Tyr Lys Pro 180 185 190 Lys Ala Gln Gly Gly Val Asn Leu Glu Asp Ser Gln Leu Val Pro Asp 195 200 205 Ser Arg Gly Gly Phe Phe Tyr Asp Val Phe Lys Glu Lys Lys Leu Thr 210 215 220 Asn Arg Ile Phe Ser Gly Ser Tyr Glu Arg Phe Gly Glu Asp Val Arg 225 230 235 240 Thr Val Leu Glu Ser Lys Gly Leu Leu Gly Leu Thr Tyr Arg Thr Leu 245 250 255 Gly Tyr Ala Thr His Ile Val Thr Val Trp Gly Ala Glu Tyr Asp Asn 260 265 270 Gln Gly Lys Ile Lys Ala Val Tyr Ile Thr Asp Ser Asp Asp Gln Gln 275 280 285 Glu Gln Ile Gly Leu Lys Arg Met Gly Ile Thr Arg Asp Ala Ser Gly 290 295 300 Asn Pro Arg Leu Asn Asn His Met Lys Asn Asn Ser Ala Gly Ala Leu 305 310 315 320 Leu Asp Tyr Val His Thr Ile Arg Leu Gly Gln Asp Leu Trp Glu 325 330 335 <210> 6 <211> 457 <212> PRT <213> Artificial <220> <223> rIdeSsuis_homolog <400> 6 Met Ala His His His His His His Val Gly Thr Gly Ser Asn Asp Asp 1 5 10 15 Asp Asp Lys Ser Pro Asp Pro Val Val Thr Gly Val Asn Glu Ile Ile 20 25 30 Glu Glu Ser Gln Val Lys Asp Glu Val Ser Ile Glu Ser Glu Lys Asn 35 40 45 Glu Ser Leu Asp Gly Ser Asn Ile Glu Ile Val Glu Glu Ile Ala Asp 50 55 60 Asn Ile Pro Ser Pro Val Ile Ala Glu Gly Glu Val Ala Val Glu Met 65 70 75 80 Lys Val Asp Arg Gly Thr Glu Asn Val Val Ser Arg Asn Asp Thr Glu 85 90 95 Val Thr Thr Ser Glu Gln Asn Gln Ile Glu Val Thr Glu Thr Lys Glu 100 105 110 Ile Leu Asn Gln Thr Ser Tyr Gln Thr Glu Ser Gly Glu Gln Arg Gln 115 120 125 Ile Ile Trp Ala His Gly Ile Thr Pro Pro Ala Met Glu Gln Ser Gly 130 135 140 Gly Phe Val Lys Glu Lys Tyr Gly Asp Tyr Leu Asn Tyr Thr Ala Pro 145 150 155 160 Phe Glu Ala Gly Lys Gly Tyr Tyr Asp Thr Asn Lys Ser Leu Asn Ala 165 170 175 Ser Phe Ile Asp Leu Asn Leu Cys Phe Ala Ala Val Ser Ser Asn Met 180 185 190 Val His Trp Trp Leu Glu Gln Asn Ser Ser Tyr Val Glu Arg Tyr Leu 195 200 205 Lys Glu Lys Lys Gly Thr Val Asn Val Glu Glu Asn Tyr Ala Ile Thr 210 215 220 Asp Leu Arg Arg Tyr Ile Asn Ser Phe Gln Asn Gln Gln Asn Ser Arg 225 230 235 240 Val Phe Asp Met Phe Lys Thr Tyr Tyr Gly Tyr Arg Thr Asn Gly Phe 245 250 255 Val Ser Asp Ala Leu Val Asp Leu Phe Ile Asn Gly Tyr Lys Pro Lys 260 265 270 Ala Gln Gly Gly Val Asn Leu Glu Asp Ser Gln Leu Val Pro Asp Ser 275 280 285 Arg Gly Gly Phe Phe Tyr Asp Val Phe Lys Glu Lys Lys Leu Thr Asn 290 295 300 Arg Ile Phe Ser Gly Ser Tyr Glu Arg Phe Gly Glu Asp Val Arg Thr 305 310 315 320 Val Leu Glu Ser Lys Gly Leu Leu Gly Leu Thr Tyr Arg Thr Leu Gly 325 330 335 Tyr Ala Thr His Ile Val Thr Val Trp Gly Ala Glu Tyr Asp Asn Gln 340 345 350 Gly Lys Ile Lys Ala Val Tyr Ile Thr Asp Ser Asp Asp Gln Gln Glu 355 360 365 Gln Ile Gly Leu Lys Arg Met Gly Ile Thr Arg Asp Ala Ser Gly Asn 370 375 380 Pro Arg Leu Asn Asn His Met Lys Asn Asn Ser Ala Gly Ala Leu Leu 385 390 395 400 Asp Tyr Val His Thr Ile Arg Leu Gly Gln Asp Leu Trp Glu Glu Tyr 405 410 415 Phe Asn Pro Leu Ala Lys Ala Cys Arg Ser Thr Ser Leu Arg Pro His 420 425 430 Ser Ser Leu Val Lys Lys Pro Leu Leu Arg Asn Leu Asn Ala Ser Thr 435 440 445 Trp Thr Arg Leu Leu Ala Gln Leu Asn 450 455 <210> 7 <211> 1016 <212> PRT <213> Artificial <220> <223> rIdeSsuisB2 <400> 7 Met Ala His His His His His His Val Gly Thr Gly Ser Asn Asp Asp 1 5 10 15 Asp Asp Lys Ser Pro Asp Pro Val Val Thr Gly Val Asn Glu Ile Ile 20 25 30 Glu Glu Ser Gln Val Lys Asp Glu Val Ser Ile Glu Ser Glu Lys Asn 35 40 45 Glu Ser Leu Asp Gly Ser Asn Ile Glu Ile Val Glu Glu Ile Ala Asp 50 55 60 Asn Ile Pro Ser Pro Val Ile Ala Glu Gly Glu Val Ala Val Glu Met 65 70 75 80 Lys Val Asp Arg Gly Thr Glu Asn Val Val Ser Arg Asn Asp Thr Glu 85 90 95 Val Thr Thr Ser Glu Gln Asn Gln Ile Glu Val Thr Glu Thr Lys Glu 100 105 110 Ile Leu Asn Gln Thr Ser Tyr Gln Thr Glu Ser Gly Glu Gln Arg Gln 115 120 125 Ile Ile Trp Ala His Gly Ile Thr Pro Pro Ala Met Glu Gln Ser Gly 130 135 140 Gly Phe Val Lys Glu Lys Tyr Gly Asp Tyr Leu Asn Tyr Thr Ala Pro 145 150 155 160 Phe Lys Ala Gly Lys Gly Tyr Tyr Asp Thr Asn Lys Ser Leu Asn Ala 165 170 175 Ser Phe Ile Asp Leu Asn Leu Cys Phe Ala Ala Val Ser Ser Asn Met 180 185 190 Val His Trp Trp Leu Glu Gln Asn Ser Ser Tyr Val Glu Arg Tyr Leu 195 200 205 Lys Glu Lys Lys Gly Thr Val Asn Val Gly Glu Asn Tyr Ala Ile Thr 210 215 220 Asp Leu Arg Arg Tyr Ile Asp Ser Phe Gln Asp Gln Gln Asn Ser Arg 225 230 235 240 Val Phe Asp Met Phe Lys Thr Tyr Tyr Gly Tyr Arg Thr Asn Gly Phe 245 250 255 Val Ser Asp Ala Leu Val Asp Leu Phe Ile Asn Gly Tyr Lys Pro Lys 260 265 270 Val Gln Gly Gly Val Asn Leu Glu Asp Ser Gln Leu Val Pro Asp Ser 275 280 285 Arg Gly Gly Phe Phe Tyr Asp Val Phe Lys Glu Lys Lys Leu Thr Asn 290 295 300 Arg Ile Phe Ser Gly Ser Tyr Glu Arg Phe Gly Glu Asp Val Arg Thr 305 310 315 320 Val Leu Glu Ser Lys Gly Leu Leu Gly Leu Thr Tyr Arg Thr Leu Gly 325 330 335 Tyr Ala Thr His Ile Val Thr Val Trp Gly Ala Glu Tyr Asp Asn Gln 340 345 350 Gly Lys Ile Arg Ala Val Tyr Ile Thr Asp Ser Asp Asp Gln Gln Glu 355 360 365 Gln Ile Gly Leu Lys Arg Met Gly Ile Thr Arg Asp Ala Ser Gly Asn 370 375 380 Pro Arg Leu Asn Asn His Val Lys Asn Asn Ser Ala Gly Ala Leu Leu 385 390 395 400 Asp Tyr Val His Thr Ile Arg Leu Gly Gln Asp Leu Trp Glu Glu Tyr 405 410 415 Phe Asn Pro Phe Ala Lys Ala Lys Glu Ile Ala Ser Gln Ile Leu Ala 420 425 430 Asp Arg Lys Lys Ala Leu Val Leu Ser Ile Gln Gly Gln Ser Glu Leu 435 440 445 Pro Glu Ser Met Arg Leu Ile Tyr Leu Glu Lys Leu Asn Asn Leu Tyr 450 455 460 Asn Gln Gly Ile Leu Ser Ile Gln Lys Thr Glu Ser Ser Glu Met Leu 465 470 475 480 Ser Gly Ala Leu Glu Asn Gly Leu Asn Ser Leu Lys Ser Leu Asp Phe 485 490 495 Pro Ile Ser Glu Val Gly Asn Ala Leu Ala Pro Asp Leu Ser Val Gly 500 505 510 Asp Arg Ser Thr Val Ser Asp Val Asp Ser Leu Ser Ser Gln Glu Thr 515 520 525 Ser Ser Thr Asn Leu Glu Ala Asp Thr Glu Asn Ala Gly Ile Ile Ala 530 535 540 Asp Gly Thr Asn Gln Leu His Phe Pro Val Glu Ala Gln Thr Thr Ser 545 550 555 560 Ser Val Glu Ala Glu Gly Asp Asn Val Phe Glu Gln Glu Ala Asp Thr 565 570 575 Leu Pro Ile Ile Ile Glu Asn Lys Asp Glu Phe Gly Ser Glu Leu Ser 580 585 590 Gly Asn Met Gln Thr Ser Glu Thr Asp Ser Leu Val Val Ala Val Glu 595 600 605 Glu Asp Val Lys Asn Asp Glu Val Asp Gln Val Glu Lys Leu Leu Glu 610 615 620 Ser Glu Lys Val Glu Asn Gln Ser Ser Glu Leu Leu Ser Asp Thr Leu 625 630 635 640 Ile Val Glu Gly Ala Asn Asp Lys Glu Glu Asp Arg Val Glu Ala Val 645 650 655 Val Ser Glu Gln Pro Asp Ser Ile Pro His Gln Asn Val Glu Ile Ser 660 665 670 Pro Val Glu Pro Met Asn Val Glu Thr Glu Ser Val Val Thr Pro Ile 675 680 685 Asn Asp Ala Ala Thr Pro His Gly Phe Pro Met Tyr Ile Asp Asn Ser 690 695 700 Val Thr Glu Ser Val Ala Thr Pro Leu Glu Lys Asp Ser Ile Gln Ala 705 710 715 720 Gly Glu Thr Glu Ile Ala Glu Pro Thr Ser Ser Glu Ser Thr Ser Val 725 730 735 Glu Ala Glu Leu Val Asp Asn Ser Glu Ile His Ser Ala Thr Ser Ser 740 745 750 Val Thr Pro Arg Gly Ser Ser Ala Tyr Ala Asp Ser Ser Thr Thr Glu 755 760 765 Ser Val Ala Thr Leu Leu Glu Lys Asp Ser Ile Gln Ala Gly Glu Thr 770 775 780 Glu Ile Ala Glu Pro Thr Ser Ser Lys Ser Thr Asn Val Glu Ala Ala 785 790 795 800 Ser Val Asp Asn Ser Glu Ile His Ala Asp Thr Ser Leu Thr Ala Val 805 810 815 Ser Ser Val Asn Leu Asp Asn Pro Val Ile Glu Pro Val Ala Ile Pro 820 825 830 Leu Ile Gly Ser Lys Arg Asp Thr Asn Ala Glu Val Glu Val Ser Ser 835 840 845 Leu Ser Lys Arg Glu Val Arg Lys Pro Asn Thr Glu Gly Leu Ile Ser 850 855 860 Val Gln Ser Lys Val Ile Lys Lys Glu Leu Leu Glu Ser Ser Leu Val 865 870 875 880 Glu Ala Gly Ser Pro Leu Leu Glu Ala Thr Ile Ala Gln Ser Ser Asn 885 890 895 Ser Asn Ser Thr Glu Ile Gly Met Ser Tyr Gln Asn Thr Val Leu Leu 900 905 910 Glu Ser Asn Asn Thr Glu Arg Gln Val Ser Lys Ala Glu Ile Val Ile 915 920 925 Glu His Lys Glu Thr Glu Leu Val Glu Thr Val Ser Ser Ala Ser Glu 930 935 940 Pro Val Val Leu Val Glu Asn Ile Ser Gln Thr Ser Asn Asn Thr Ile 945 950 955 960 Glu Ser Gly Lys Asn Met Gly Val Gln Ser Gln Ala Gly Ala Lys Gln 965 970 975 Ile Leu Gly Ile Glu Gln Ser Ser Lys Val Ser Thr Pro Thr Ser Arg 980 985 990 Gln Ile Met Gly Val Gly Leu Leu Thr Leu Val Leu Gly Ser Ala Leu 995 1000 1005 Gly Leu Leu Lys Lys Arg Arg Lys 1010 1015 <210> 8 <211> 1374 <212> DNA <213> Artificial <220> <223> Nucleotide sequence encoding rIdeSsuis homolog <400> 8 atggcacatc accaccacca tcacgtgggt accggttcga atgatgacga cgacaagagt 60 ccggatccag ttgttacagg agtgaatgaa ataattgaag aatcacaagt caaggatgag 120 gtatctattg aatcagaaaa aaatgaatcc ctagatggtt ctaatattga aattgtagag 180 gaaatagcag acaacatccc atcacctgtt atcgctgaag gggaagtagc ggtagagatg 240 aaagttgaca gagggaccga gaatgtagtt tctagaaatg atacagaagt tacgacgagc 300 gagcaaaatc agatagaggt tactgagaca aaagaaattt tgaatcagac cagttatcaa 360 acggagtg gcgagcaacg acaaattata tgggcccatg gatttactcc tcctgcaatg 420 gaacaaagcg gtggttttgt aaaggaaag tatggagact attackaacta tacagcgcca 480 tttgaggctg gaaaaggcta ctatgatacc addaagagtc tgaatgctc atttattgac 540 cttaatcttt gtttgcagc tgtgtctca aacatggtac attggtggtt ggaacagaat 600 agttcctatg ttgagcgata tctcaagaa aaaaagggta cagtaaatgt tgagaaaac 660 tacgcaataa cggacttacg gcgctatatt attcattcc aaatcaca aaatagtcga 720 gtttttgata tgttcaaac ttactatggt tatcgtacaa atggttttgt atcagatgcc 780 ttggttgact tgtttaa cggatataaa cctaggcac agggcggtgt caatctggaa 840 gatagccagt tagtaccaga tagtaggtt ggctttttct acgacgtttt caagagaaa 900 aaactgacaa atcgaatttt tagtggtagt tatgagcggt tggtgagga tgttcgaact 960 gttttggaaa gcaaggatt actcggctta acttatagaa cattaggtta tgcaacgcat 1020 attgtgacgg tatggggtgc tgagtacgac atcaggta agatttaggc tgtctatatc 1080 acagattctg atgatcaaca agaacaaatt ggtttgaagc gtatgggaat cactcgtgat 1140 gcttccggaa atccacgttt gaataatcat atgaaaaata attcagctgg agcgcttttg 1200 gattatgtcc atacaatccg tctgggtcaa gacttatggg aagaatattt caatccgctt 1260 gcaaaagcct gcaggtcgac aagcttgcgg ccgcactcga gtctggtaaa gaaaccgctg 1320 ctgcgaaatt tgaacgccag cacatggact cgtctactag cgcagcttaa ttaa 1374 <210> 9 <211> 3051 <212> DNA <213> Artificial <220> <223> Nucleotide sequence encoding rIdeSsuisB2 <400> 9 atggcacatc accaccacca tcacgtgggt accggttcga atgatgacga cgacaagagt 60 ccggatccag ttgttacagg agtgaatgaa ataattgaag aatcacaagt caaggatgag 120 gtatctattg aatcagaaaa aaatgaatcc ctagatggtt ctaatattga aattgtagag 180 gaaatagcag acaacatccc atcacctgtt atcgctgaag gggaagtagc ggtagagatg 240 aaagttgaca gagggaccga gaatgtagtt tctagaaatg atacagaagt tacgacgagc 300 gagcaaatc agatagaggt tactgagaca aaagaaattt tgaatcagac cagttatca 360 acggagtg gcgagcaacg acaaattata tgggcccatg gatttactcc tcctgcaatg 420 gaacaaagcg gtggttttgt aaaggaaag tatggagact attackaacta tacagcgcca 480 tttaggcag gaaaaggcta ttatgatacc aaaaagtc tcaatgctc atttattgac 540 cttaacctat gtttgcagc cgtgtcttcc aacatggtac attggtggtt ggaacagaat 600 agttcctatg ttgagcgata tctcaagaa aaaaagggta cagtaaatgt tggggaaac 660 tatgcaataa cggacctacg tcgctatatt gattcgttcc aggatcagca aaatagtcga 720 gtctttgata tgttcaaac ttactacggt tatcgtacaa atggttttgt gtcagatgcc 780 ctagttgact tgtttattaa tggatataaa cctaagtac agggtggtgt caatctggaa 840 gatagccagt tagtaccaga tagtaggtt ggctttttct acgacgtttt caagagaaa 900 aaactgacaa atcgtatttt tagtggtagc tatgagcgtt tggtgagga tgttcgaact 960 gttttggaga gcaaggatt actcggtcta acttatagaa cattaggcta tgcaacgcat 1020 attgtgacgg tatggggtgc tgagtacgat aatcaggta agattagggc tgtctatatc 1080 actgattccg atgatcaaca agacaatt ggtttgaagc gtatgggaat cactcgtgat 1140 gcttctggaa atccgcgttt gataatcat gtgaaaaata attcagctgg gggcgctttg 1200 gattatgtcc atacaatccg tcttggtca gacttatggg agaatattt caatccgttc 1260 gcaaaagcca aagaatagc tagtcagata ctagctgata gaaagaaggc gttggttctg 1320 tctattcaag gatactga attgccagaa tcaatgcggc tgatttact tgaaaaacta 1380 aaatctct ataatcagg gattctatct attcaaaga cagaagttc tgagatgcta 1440 agtggtgcat tggaaatgg ttaatagt ttaagagtt tagattttcc tatttcaga 1500 gttggaaatg ctttggcacc agatttatca gtaggtgatc gctcaacggt ttcagatgtt 1560 gattctctat catctcaaga aacaagttcc acaatttgg aagcagacac agagaatgca 1620 ggtattattg cagatggtac caatcattg cattttccag tggaggccca acgacatct 1680 tcagtagagg ctgagggaga taatgtttt gaacagagg cagatacatt accaatatt 1740 attgaaaaca aggatgaatt tggttcagaa ctatcaggaa acatgcaac gtcagaaacg 1800 gattcgctag tagtagctt tgaagagat gtgaaaaatg atgaggtaga ccaagttgaa 1860 aagctcttg aatcagaaaa agttgaaat cagagttcgg aacttctgtc agacacccta 1920 atcgtagagg gtgcaatga caagagaa gatgagtgg aggcggttgt ttctgaaca 1980 ccagactcaa taccacatca aaatgtagaa atctctcctg tagaccaat gatgtcgaa 2040 actgaatctg tggtcactcc tattaatgat gcagctactc ctcatggttt cccgatgtat 2100 atagataatt ccgtactga atctgtagct actccacttg aaaagactc cattcaagcc 2160 ggagagacag aaattgcaga accaacctcg agcgaatcaa ctagtgttga agctgaactt 2220 gtcgacaatt ctgaaatcca ttcagctacc tctcagtta ctccccgtgg ttcctcggca 2280 tatgcagata gttccacac tgaatctgta gctactctgc ttgaaaaga ctccattcag 2340 gctggagaga cagaaattgc agaaccaacc tcgagcaat cactaatgt cgaagctgca 2400 tctgtcgaca attctgaaat tcatgcagat accctctaa ctgctgtttc atcagtcaat 2460 ctggataatc cagtgattga accagtagct atccccctta tcggttctaa gagggacacg 2520 aatgcagaag tggaagtttc ttcattatcg aaaagagagg ttagaaaacc aaatactgaa 2580 gggctaatct ctgttcaatc aaaagttatt aagaaagaat tgctagaatc aagcttagta 2640 gaagcagggt ctccattgct agaagccacc attgctcagt cttcaaactc aaatagtact 2700 gagataggta tgagctatca gaatactgtg ttattagagt ctaataatac agagcgtcag 2760 gtgtctaaag cagaaattgt tatagaacac aaggagacag agttagttga aacggtttca 2820 tctgcttctg agcctgtagt gctcgtagaa aatatctcac aaacctcaaa taatactatt 2880 gaatctggta agaatatggg agttcaatct caagcaggtg caaaacaaat tttaggcata 2940 gaacaatctt cgaaagtaag tacacctact tcaagacaga ttatgggagt cggtctattg 3000 actcttgttc ttggtagtgc tttaggtttg ttaaagaaaa gacgtaagta a 3051 <210> 10 <211> 335 <212> PRT <213> Artificial <220> <223> Amino acids 91 - 425 of WP_044671938 <400> 10 Met Lys Val Asp Arg Gly Thr Glu Asn Val Val Ser Arg Asn Asp Thr 1 5 10 15 Glu Val Thr Thr Ser Glu Gln Asn Gln Ile Glu Val Thr Glu Thr Lys 20 25 30 Glu Ile Leu Asn Gln Thr Ser Tyr Gln Thr Glu Ser Gly Glu Gln Arg 35 40 45 Gln Ile Ile Trp Ala His Gly Ile Thr Pro Pro Ala Met Glu Gln Ser 50 55 60 Gly Gly Phe Val Lys Glu Lys Tyr Gly Asp Tyr Leu Asn Tyr Thr Ala 65 70 75 80 Pro Phe Lys Ala Gly Lys Gly Tyr Tyr Asp Thr Asn Lys Ser Leu Asn 85 90 95 Ala Ser Phe Ile Asp Leu Asn Leu Cys Phe Ala Ala Val Ser Ser Asn 100 105 110 Met Val His Trp Trp Leu Glu Gln Asn Ser Ser Tyr Val Glu Arg Tyr 115 120 125 Leu Lys Glu Lys Lys Gly Thr Val Asn Val Gly Glu Asn Tyr Ala Ile 130 135 140 Thr Asp Leu Arg Arg Tyr Ile Asp Ser Phe Gln Asp Gln Gln Asn Ser 145 150 155 160 Arg Val Phe Asp Met Phe Lys Thr Tyr Tyr Gly Tyr Arg Thr Asn Gly 165 170 175 Phe Val Ser Asp Ala Leu Val Asp Leu Phe Ile Asn Gly Tyr Lys Pro 180 185 190 Lys Val Gln Gly Gly Val Asn Leu Glu Asp Ser Gln Leu Val Pro Asp 195 200 205 Ser Arg Gly Gly Phe Phe Tyr Asp Val Phe Lys Glu Lys Lys Leu Thr 210 215 220 Asn Arg Ile Phe Ser Gly Ser Tyr Glu Arg Phe Gly Glu Asp Val Arg 225 230 235 240 Thr Val Leu Glu Ser Lys Gly Leu Leu Gly Leu Thr Tyr Arg Thr Leu 245 250 255 Gly Tyr Ala Thr His Ile Val Thr Val Trp Gly Ala Glu Tyr Asp Asn 260 265 270 Gln Gly Lys Ile Lys Ala Val Tyr Ile Thr Asp Ser Asp Asp Gln Gln 275 280 285 Glu Gln Ile Gly Leu Lys Arg Met Gly Ile Thr Arg Asp Ala Ser Gly 290 295 300 Asn Pro Arg Leu Asn Asn His Met Lys Asn Asn Ser Ala Gly Ala Leu 305 310 315 320 Leu Asp Tyr Val His Thr Ile Arg Leu Gly Gln Asp Leu Trp Glu 325 330 335 <210> 11 <211> 335 <212> PRT <213> Artificial <220> <223> Amino acids 91 - 425 of WP_002935529 <400> 11 Met Lys Val Asp Arg Gly Thr Glu Asn Val Val Ser Arg Asn Asp Thr 1 5 10 15 Glu Val Thr Thr Ser Glu Gln Asn Gln Ile Glu Val Thr Glu Thr Lys 20 25 30 Glu Ile Leu Asn Gln Thr Ser Tyr Gln Thr Glu Ser Gly Glu Gln Arg 35 40 45 Gln Ile Ile Trp Ala His Gly Ile Thr Pro Pro Ala Met Glu Gln Ser 50 55 60 Gly Gly Phe Val Lys Glu Lys Tyr Gly Asp Tyr Leu Asn Tyr Thr Ala 65 70 75 80 Pro Phe Lys Ala Gly Lys Gly Tyr Tyr Asp Thr Asn Lys Ser Leu Asn 85 90 95 Ala Ser Phe Ile Asp Leu Asn Leu Cys Phe Ala Ala Val Ser Ser Asn 100 105 110 Met Val His Trp Trp Leu Glu Gln Asn Ser Ser Tyr Val Glu Arg Tyr 115 120 125 Leu Lys Glu Lys Lys Gly Thr Val Asn Val Gly Glu Asn Tyr Ala Ile 130 135 140 Thr Asp Leu Arg Arg Tyr Ile Asp Ser Phe Gln Asp Gln Gln Asn Ser 145 150 155 160 Arg Val Phe Asp Met Phe Lys Thr Tyr Tyr Gly Tyr Arg Thr Asn Gly 165 170 175 Phe Val Ser Asp Ala Leu Val Asp Leu Phe Ile Asn Gly Tyr Lys Pro 180 185 190 Lys Val Gln Gly Gly Val Asn Leu Glu Asp Ser Gln Leu Val Pro Asp 195 200 205 Ser Arg Gly Gly Phe Phe Tyr Asp Val Phe Lys Glu Lys Lys Leu Thr 210 215 220 Asn Arg Ile Phe Ser Gly Ser Tyr Glu Arg Phe Gly Glu Asp Val Arg 225 230 235 240 Thr Val Leu Glu Ser Lys Gly Leu Leu Gly Leu Thr Tyr Arg Thr Leu 245 250 255 Gly Tyr Ala Thr His Ile Val Thr Val Trp Gly Ala Glu Tyr Asp Asn 260 265 270 Gln Gly Lys Ile Arg Ala Val Tyr Ile Thr Asp Ser Asp Asp Gln Gln 275 280 285 Glu Gln Ile Gly Leu Lys Arg Met Gly Ile Thr Arg Asp Ala Ser Gly 290 295 300 Asn Pro Arg Leu Asn Asn His Val Lys Asn Asn Ser Ala Gly Ala Leu 305 310 315 320 Leu Asp Tyr Val His Thr Ile Arg Leu Gly Gln Asp Leu Trp Glu 325 330 335 <210> 12 <211> 335 <212> PRT <213> Artificial <220> <223> Amino acids 92 - 426 of WP_015647040 <400> 12 Met Lys Val Asp Arg Gly Thr Glu Asn Val Val Ser Arg Asn Asp Lys 1 5 10 15 Glu Val Thr Thr Ser Glu Lys Asn Gln Ile Glu Val Thr Glu Thr Lys 20 25 30 Glu Ile Leu Asn Gln Thr Ser Tyr Gln Thr Glu Ser Gly Glu Gln Arg 35 40 45 Gln Ile Ile Trp Ala His Gly Ile Thr Pro Pro Ala Met Glu Gln Ser 50 55 60 Gly Gly Phe Val Lys Glu Lys Tyr Gly Asp Tyr Leu Asn Tyr Thr Ala 65 70 75 80 Pro Phe Glu Ala Gly Lys Gly Tyr Tyr Asp Thr Asn Lys Ser Leu Asn 85 90 95 Ala Ser Phe Ile Asp Leu Asn Leu Cys Phe Ala Ala Val Ser Ser Asn 100 105 110 Met Val His Trp Trp Leu Glu Gln Asn Ser Ser Tyr Val Glu Arg Tyr 115 120 125 Leu Lys Glu Lys Asn Ser Thr Val Asn Val Gly Glu Asn Tyr Ala Ile 130 135 140 Thr Asp Leu Arg Arg Tyr Ile Asn Ser Phe Gln Asn Gln Gln Asn Ser 145 150 155 160 Arg Val Phe Asp Met Phe Lys Thr Tyr Tyr Gly Tyr Arg Thr Asn Gly 165 170 175 Phe Val Ser Asp Ala Leu Val Asp Leu Phe Ile Asn Gly Tyr Lys Pro 180 185 190 Lys Ala Gln Gly Gly Val Asn Leu Glu Asp Ser Gln Leu Val Pro Asp 195 200 205 Ser Arg Gly Gly Phe Phe Tyr Asp Val Phe Lys Glu Lys Lys Leu Thr 210 215 220 Asn Arg Ile Phe Ser Gly Ser Tyr Glu Arg Phe Gly Glu Asp Val Arg 225 230 235 240 Thr Val Leu Glu Ser Lys Gly Leu Leu Gly Leu Thr Tyr Arg Thr Leu 245 250 255 Gly Tyr Ala Thr His Ile Val Thr Val Trp Gly Ala Glu Tyr Asp Asn 260 265 270 Gln Gly Lys Ile Lys Ala Val Tyr Ile Thr Asp Ser Asp Asp Gln Gln 275 280 285 Glu Gln Ile Gly Leu Lys Arg Met Gly Ile Thr Arg Asp Ala Ser Gly 290 295 300 Asn Pro Arg Leu Asn Asn His Val Lys Asn Asn Ser Ala Gly Ala Leu 305 310 315 320 Leu Asp Tyr Val His Thr Ile Arg Leu Gly Gln Asp Leu Trp Glu 325 330 335 <210> 13 <211> 335 <212> PRT <213> Artificial <220> <223> Amino acids 92 - 426 of WP_023370787 <400> 13 Met Lys Ser Asp Asn Gly Asp Glu Asn Ala Val Ser Arg Asp Asp Ser 1 5 10 15 Glu Val Thr Thr Asn Glu Gln Asn Gln Ile Glu Val Thr Glu Thr Lys 20 25 30 Glu Ile Leu Asn Gln Thr Ser Tyr Gln Thr Glu Ser Gly Glu Gln Arg 35 40 45 Gln Ile Ile Trp Ala His Gly Ile Thr Pro Pro Ala Met Glu Gln Ser 50 55 60 Gly Gly Phe Val Lys Glu Lys Tyr Gly Asp Tyr Leu Asn Tyr Thr Ala 65 70 75 80 Pro Phe Glu Ala Gly Lys Gly Tyr Tyr Asp Thr Asn Lys Ser Leu Asn 85 90 95 Ala Ser Phe Ile Asp Leu Asn Leu Cys Phe Ala Ala Val Ser Ser Asn 100 105 110 Met Val His Trp Trp Leu Glu Gln Asn Ser Ser Tyr Val Glu Arg Tyr 115 120 125 Leu Lys Glu Lys Lys Gly Thr Val Asn Val Glu Glu Asn Tyr Ala Ile 130 135 140 Thr Asp Ile Arg Arg Tyr Ile Asn Ser Phe Gln Asn Gln Gln Asn Ser 145 150 155 160 Arg Val Phe Asp Met Phe Lys Thr Tyr Tyr Gly Tyr Arg Thr Asn Gly 165 170 175 Phe Val Ser Asp Ala Leu Val Asp Leu Phe Ile Asn Gly Tyr Lys Pro 180 185 190 Lys Ser Gln Gly Gly Val Asn Leu Glu Asp Ser His Leu Val Pro Asp 195 200 205 Ser Arg Gly Gly Phe Phe Tyr Asp Val Phe Lys Glu Lys Lys Leu Thr 210 215 220 Asn Arg Ile Phe Ser Gly Ser Tyr Glu Arg Phe Gly Glu Asp Val Arg 225 230 235 240 Thr Val Leu Glu Ser Lys Gly Leu Leu Gly Leu Thr Tyr Arg Thr Leu 245 250 255 Gly Tyr Ala Thr His Ile Val Thr Val Trp Gly Ala Glu Tyr Asp Asn 260 265 270 Gln Gly Lys Ile Lys Ala Val Tyr Ile Thr Asp Ser Asp Asp Gln Gln 275 280 285 Glu Gln Ile Gly Leu Lys Arg Met Gly Ile Thr Arg Asp Ala Ser Gly 290 295 300 Asn Pro Arg Leu Asn Asn His Met Lys Asn Asn Ser Ala Gly Ala Leu 305 310 315 320 Leu Asp Tyr Val His Thr Ile Arg Leu Gly Gln Asp Leu Trp Glu 325 330 335 <210> 14 <211> 335 <212> PRT <213> Artificial <220> <223> Amino acids 92 - 426 of WP_044678723 <400> 14 Met Asn Ser Asp Asn Gly Asp Glu Asn Val Val Ser Arg Asp Asp Ser 1 5 10 15 Glu Val Thr Thr Asn Glu Gln Asn Gln Ile Glu Val Thr Glu Thr Lys 20 25 30 Glu Ile Leu Asn Tyr Thr Ser Tyr Gln Thr Glu Ser Gly Glu Gln Arg 35 40 45 Gln Ile Val Trp Ala Tyr Gly Ile Thr Pro Pro Val Met Glu Gln Lys 50 55 60 Gly Gly Phe Val Lys Glu Lys Tyr Gly Asp Tyr Leu Asn Tyr Thr Ala 65 70 75 80 Pro Phe Glu Ala Gly Lys Gly Tyr Tyr Asp Thr Asn Lys Ser Leu Asn 85 90 95 Ala Ser Phe Ile Asp Leu Asn Leu Cys Phe Ala Ala Val Ser Ser Asn 100 105 110 Met Val His Trp Trp Leu Glu Gln Asn Ser Ser Tyr Val Glu Arg Tyr 115 120 125 Leu Lys Glu Lys Lys Gly Thr Val Asn Val Glu Glu Asn Tyr Ala Ile 130 135 140 Thr Asp Leu Arg Arg Tyr Ile Asn Ser Phe Gln Asn Gln Gln Asn Ser 145 150 155 160 Arg Val Phe Asp Met Phe Lys Thr Tyr Tyr Gly Tyr Arg Thr Asn Gly 165 170 175 Phe Val Ser Asp Ala Leu Val Asp Leu Phe Ile Asn Gly Tyr Lys Pro 180 185 190 Lys Ala Gln Gly Gly Val Asn Leu Glu Asp Ser Gln Leu Val Pro Asp 195 200 205 Ser Arg Gly Gly Phe Phe Tyr Asp Val Phe Lys Glu Lys Lys Leu Thr 210 215 220 Asn Arg Ile Phe Ser Gly Ser Tyr Glu Arg Phe Gly Glu Asp Val Arg 225 230 235 240 Thr Val Leu Glu Ser Lys Gly Leu Leu Gly Leu Thr Tyr Arg Thr Leu 245 250 255 Gly Tyr Ala Thr His Ile Val Thr Val Trp Gly Ala Glu Tyr Asp Asn 260 265 270 Gln Gly Lys Ile Lys Ala Val Tyr Ile Thr Asp Ser Asp Asp Gln Gln 275 280 285 Glu Gln Ile Gly Leu Lys Arg Met Gly Ile Thr Arg Asp Ala Ser Gly 290 295 300 Asn Pro Arg Leu Asn Asn His Met Lys Asn Asn Ser Ala Gly Ala Leu 305 310 315 320 Leu Asp Tyr Val His Thr Ile Arg Leu Gly Gln Asp Leu Trp Glu 325 330 335

Claims

1. Use of a vaccine composition in the preparation of a medicament for preventing, immunomodulating or treating Streptococcus suis infection in pigs, the vaccine composition comprising an effective amount of an IdeSsuis protein that induces IdeSsuis-specific IgG antibodies with neutralizing activity, or a vector into which a polynucleotide encoding an IdeSsuis protein that induces IdeSsuis-specific IgG antibodies with neutralizing activity is inserted; and at least one pharmaceutical carrier, diluent or adjuvant; The protein is an amino acid sequence of SEQ ID NO: 5, or SEQ ID NO: 1 with amino acids 1 to 34 missing.

2. The use according to claim 1, characterized in that The IdeSsuis protein is part of a fusion protein with at least another protein.

3. The use according to claim 1 or 2, characterized in that The polynucleotide is DNA or RNA.

4. The use according to claim 1, characterized in that The polynucleotide is integrated into a vector, wherein the polynucleotide is operably linked to the expression control region of the vector, and the vector is an expression vector, and the expression vector is a plasmid or a viral vector.

5. The use according to claim 1, wherein the protein is the only active or immunogenic component.

6. The use according to claim 1, characterized in that: The vaccine composition is provided in a physiologically administrable form for intramuscular, intravenous, subcutaneous or cutaneous injection or mucosal application.

7. The use according to claim 1, wherein the treatment comprises at least one time.

8. The use according to claim 1, wherein the treatment comprises at least two immunizations.

9. The use according to claim 1, wherein the dosage of protein administered to each pig is 0.05-2.0 mg.

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