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

A technology of Streptococcus suis and surface protein, applied in the directions 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: 2009-12-30
MICROBE EPIDEMIC DISEASE INST OF PLA MILITARY MEDICAL ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

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 and purification of Streptococcus suis type 2 surface protein SSU98_1675 and antibody preparation

[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 2Chinese Isolates.PLoS ONE 2(3) :E315]; Escherichia coli DH5a and BL21, both of which are international standard strains; the expression vector pET32a(+) is a product of Novagen.

[0020] 1.2 Enzymes and reagents

[0021] Restriction enzymes such as EcoRI and XhoI, as well as Taq enzymes, dNTPs, DL15000 molecular weight standards and other PCR reagents used in PCR are all products of Bao Bioengineering (Dalian) Co., Ltd.; DNA recovery kits are products of Tianwei Company; nickel affinity chromatography columns It is a product of GE; the plasmid extraction kit is a product of V-gene; Freund’s complete adjuv...

Embodiment 2

[0048] Example 2 The immunogenicity of Streptococcus suis type 2 surface protein SSU98_1675 and antibody-mediated opsonization phagocytosis test

[0049] 1. Material method: The material is the same as in Example 1.

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

[0051] 1.2 SSU98_1675 antibody-mediated opsonization

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

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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 the Escherichia 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 (Streptococcus suis) belongs to the R group of the Ransali classification method. There are 35 serotypes. Streptococcus suis type 2 is the most prevalent and the most pathogenic. Since Denmark first reported human infection with Streptococcus suis in 1968, some countries in Europe and the United States and Hong Kong have reported cases of Streptococcus suis since the 1970s, mainly meningitis and toxic shock syndrome (TSS) Rare, no outbreaks have been seen. In my country, there were two outbreaks of human Streptococcus suis type 2 in Jiangsu and Sichuan in 1998 and 2005. Different from the characteristics of meningitis in foreign cases, the clinical manifestations are shock and meningitis. Most cases died of toxic shock s...

Claims

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Application Information

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Patent Type & Authority Applications(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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