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

A surface protein, Streptococcus suis technology, applied in the field of bacterial proteins, can solve problems such as difficulties in identifying immunogenic proteins, false positive reactions, and inability to achieve high-throughput screening

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

AI Technical Summary

Problems solved by technology

At present, there is still a lack of powerful means to obtain immunogenic proteins. It is difficult to identify immunogenic proteins by traditional biological methods. It is necessary to construct a gene library and then screen with the corresponding serum. This method takes a long time and cannot be realized. High-throughput screening with potential for false positives

Method used

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

Examples

Experimental program
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Embodiment 1

[0021] Example 1 Expression and purification of Streptococcus suis type 2 surface protein 0186, immunogenicity and antibody-mediated opsonophagocytosis test

[0022] 1 Materials and methods

[0023] 1.1 Strains and plasmids

[0024] 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.

[0025] 1.2 Enzymes and Reagents

[0026] 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-gen...

Embodiment 2 86

[0066] The preparation process of embodiment two 186 colloidal gold detection test strips

[0067] 1. Antibody Processing

[0068] The immune serum obtained by immunizing rats with 186 and New Zealand rabbits with Streptococcus suis was purified by caprylic acid-ammonium sulfate precipitation or protein A column, dialyzed overnight with 0.01M PBS, centrifuged at 10,000r / min for 30min, and collected the supernatant to measure the concentration. use.

[0069] 2. Preparation of colloidal gold (sodium citrate reduction method)

[0070] HAuCl 4 First make a 1% aqueous solution, take 100ml of pure water and heat it to boiling. Add 1ml of 1% HAuCl while stirring 4 and 1.5 ml of trisodium citrate in water. Continue to heat and boil for about 15 minutes. After adding the liquid, it is observed that the color turns black first, and then slowly turns into wine red. After seeing the wine red, continue heating for 3-5 minutes and then stop heating. The obtained is colloidal gold with a...

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Abstract

The invention discloses a surface protein of streptococcus suis 2-type and also discloses a preparation method and application of the protein. The protein can be used as a diagnostic target and used for developing a diagnostic reagent for the streptococcus suis 2-type. 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 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 of the ...

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 姜永强郑玉玲耿红冉张炜李文君袁媛熊国华
Owner MICROBE EPIDEMIC DISEASE INST OF PLA MILITARY MEDICAL ACAD OF SCI