Surface protein of streptococcus suis type-2, preparation method thereof and application

A technology of Streptococcus suis and surface protein, which is applied in the fields of botanical equipment and methods, biochemical equipment and methods, chemical instruments and methods, etc., can solve problems such as difficult to achieve ideal effects, and achieve the effect of broad market prospects.

Active Publication Date: 2012-11-28
MICROBE EPIDEMIC DISEASE INST OF PLA MILITARY MEDICAL ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Both endemic strains and non-endemic strains in my country express MRP, and animal experiments show that the supernatants secreted by the endemic strains do not cause pathological changes. The virulence factors such as MRP, EF, and SLY are not enough to explain the type 2 pig chain in my country. The pathogenic mechanism of cocci, using MRP as a human immune vaccine to produce antibodies to neutralize MRP may be difficult to achieve the desired effect

Method used

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  • Surface protein of streptococcus suis type-2, preparation method thereof and application
  • Surface protein of streptococcus suis type-2, preparation method thereof and application

Examples

Experimental program
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Effect test

Embodiment 1

[0016] Example 1 Expression, purification and antibody preparation of Streptococcus suis type 2 surface protein SSU98_1675

[0017] 1. Materials and Methods

[0018] 1.1 Strains and plasmids

[0019] Streptococcus suis type 2 98HAH12 strain [Chen C, Tang J, Dong W, Wang C, Feng Y, et al(2007) A Glimpse of Streptococcal Toxic Shock Syndrome from Comparative Genomics of S.suis 2 Chinese Isolates.PLoS ONE 2(3) : e315]; Escherichia coli DH5a, BL21, are international standard strains; expression vector pET32a (+) is a product of Novagen.

[0020] 1.2 Enzymes and Reagents

[0021] EcoRI, XhoI and other restriction endonucleases, as well as Taq enzymes, dNTPs, DL15000 molecular weight standards and other PCR reagents are all products of Bao Biological Engineering (Dalian) Co., Ltd.; DNA recovery kits are products of Tianwei Company; nickel affinity chromatography column It is a product of GE Company; the plasmid extraction kit is a product of V-gene Company; Freund's complete adju...

Embodiment 2

[0048] Example 2 Immunogenicity of Streptococcus suis type 2 surface protein SSU98_1675 and antibody-mediated opsonophagocytosis test

[0049] 1. Material method: material is the same as embodiment one.

[0050] 1.1 Preparation of recombinant Streptococcus suis type 2 surface protein SSU98_1675 and polyclonal antibody: see Example 1.

[0051] 1.2 Opsonophagocytosis mediated by SSU98_1675 antibody

[0052] Scrape a single colony of Streptococcus suis type 2 from the blood plate, insert into THB medium, 37°C, CO 2 After culturing in an incubator for 8 hours, transfer to fresh THB medium and culture for 8 hours to logarithmic growth phase. Take 1ml bacterial solution (about 1×10 8 CFU / ml) 13,000rpm, collect bacteria for 1min, wash with PBS and resuspend, adjust to 10 6 CFU / ml, smear blood plate, recorded as the initial bacterial count. Take 50μl (≈10 6 CFU / ml) bacteria solution, add 100 μl antibody, 15 minutes at 37°C and then put on ice for 15 minutes; add 350 μl healthy h...

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Abstract

The invention discloses a surface protein of streptococcus suis type-2, preparation method thereof and application. The surface protein of streptococcus suis type-2 is numbered as gi|146321522 in genebank; the preparation method of the surface protein comprises the following steps: cloning gene of the surface protein; connecting the gene and expression plasmid of Escherichia coli to transform theEscherichia coli; inducing expressing; separating and purifying expression product; and the like. The surface lipoprotein can be used as a diagnostic target, and a specific antibody of the protein can be detected in animals and people infected with the streptococcus suis; moreover, the surface lipoprotein is also an important protective antigen, and can be used for preparing vaccines.

Description

technical field [0001] The invention relates to a bacterial surface protein, in particular to a surface protein of Streptococcus suis type 2, and also relates to a preparation method and application of the protein. Background technique [0002] Streptococcus suis belongs to the R group of Lang's classification method and has 35 serotypes, among which Streptococcus suis type 2 is the most prevalent and most pathogenic. Since Denmark first reported human infection with Streptococcus suis in 1968, since the 1970s, some countries in Europe and the United States and Hong Kong have reported cases of Streptococcus suis, mainly meningitis, toxic shock syndrome (TSS) Rare, no outbreaks have been seen. In 1998 and 2005, there were two outbreaks of human infection with Streptococcus suis type 2 in Jiangsu and Sichuan in 1998 and 2005, which were different from the characteristics of meningitis in foreign cases, and the clinical manifestations were shock type and meningitis type. Most...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/315C12N15/31C12N15/70C12N1/21G01N33/569A61K39/09A61P31/04C12R1/19
Inventor 姜永强尉研耿红冉郑玉玲张炜李文君袁媛江华
Owner MICROBE EPIDEMIC DISEASE INST OF PLA MILITARY MEDICAL ACAD OF SCI
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