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Nucleic acid enzyme surface protein of streptococcus suis 2-type and preparation method and application thereof

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

Active Publication Date: 2013-04-17
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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  • Nucleic acid enzyme surface protein of streptococcus suis 2-type and preparation method and application thereof
  • Nucleic acid enzyme surface protein of streptococcus suis 2-type and preparation method and application thereof

Examples

Experimental program
Comparison scheme
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Embodiment 1

[0016] Example 1 Expression and purification of Streptococcus suis type 2 nuclease surface protein SSU98_1549 and antibody preparation

[0017] 1. Materials and methods

[0018] 1.1 Strains and plasmids

[0019] Streptococcus suis type 2 strain 98HAH12 [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, BL21, all of which are international standard strains; 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 PCR reagents such as Taq enzyme, dNTPs, and DL15000 molecular weight standards are 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 company; the plasmid extraction box is a product of V-gene company; Freund's com...

Embodiment 2

[0047] Example 2 Immunogenicity of Streptococcus suis type 2 nuclease surface protein SSU98_1549 and antibody-mediated opsonophagocytosis test

[0048] 1. Materials and methods: The materials are the same as those in Example 1.

[0049] 1.1 Preparation of recombinant Streptococcus suis type 2 nuclease surface protein SSU98_1549 and polyclonal antibody: see Example 1.

[0050] 1.2 SSU98_0267 antibody-mediated opsonophagy

[0051] Scrape a single colony of Streptococcus suis type 2 from the blood plate, insert it into THB medium, 37°C, CO 2 After culturing for 8 h in the incubator, the cells were transferred to fresh THB medium for 8 h to the logarithmic growth phase. Take 1ml of bacterial solution (about 1×10 8 CFU / ml) 13,000rpm, collect bacteria for 1min, wash bacteria with PBS and resuspend, adjust to 10 6 CFU / ml, coated with blood plate, recorded as the initial bacterial volume. Take 50μl (≈10 6 CFU / ml) bacterial solution was added with 100 μl antibody, 37°C for 15 min...

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Abstract

The invention discloses a nucleic acid enzyme surface protein of streptococcus suis 2-type and also discloses a preparation method and application of the protein. The number of the nucleic acid enzyme surface protein of the streptococcus suis 2-type in a genebank is gi (146321396); the preparation method comprises: cloning a nucleic acid enzyme surface protein gene; connecting the gene to colibacillus expression plasmid, and transforming colibacillus; inducing expression; and separating and purifying an expression product and other steps. The nucleic acid enzyme surface protein can be used asa diagnostic target; and a specific antibody of the protein can be detected in animals and people infected with the streptococcus suis. Simultaneously, the protein is also an important protective antigen and can be used for preparing vaccine.

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 to a preparation method and application of the protein. Background technique [0002] Streptococcus suis (Streptococcus suis), belonging to the R group of the Rankine classification, has 35 serotypes. Streptococcus suis type 2 is the most prevalent and the most pathogenic. Since Denmark first reported human infection with Streptococcus suis disease in 1968, since the 1970s, some European and American countries and Hong Kong have reported cases of Streptococcus suis, mainly meningitis and toxic shock syndrome (TSS). Rare, no outbreaks have been seen. In 1998 and 2005, two outbreaks of human infection with Streptococcus suis type 2 occurred in Jiangsu and Sichuan in 1998 and 2005. Different from foreign cases, which are mainly meningitis, the clinical manifestations are shock type and meningitis type. Most of the cases died of...

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 姜永强郝淮杰郑玉玲耿红冉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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