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Streptococcus recombination subunit vaccine and preparation method

A technology of Streptococcus suis and lysozyme, which is applied in the field of genetic engineering recombinant protein vaccines, can solve problems such as the useless genetic engineering vaccine of Streptococcus suis type 2, and achieves the effects of good immunogenicity, convenient use and enhanced protection.

Inactive Publication Date: 2013-03-20
陆承平
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Streptococcus suis type 2 currently has no genetically engineered vaccine for clinical use

Method used

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  • Streptococcus recombination subunit vaccine and preparation method
  • Streptococcus recombination subunit vaccine and preparation method
  • Streptococcus recombination subunit vaccine and preparation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] 1.1 Obtain the coding gene of Streptococcus suis type 2 MRP-HM6 fusion epitope

[0027] Streptococcus suis type 2 HA9801 strain Yao Huochun et al. [Journal of Nanjing Agricultural University, 1999, 22(2): 67~70] was isolated and identified from a diseased pig with a streptococcus suis outbreak in Jiangsu in 1998, and was identified in August 2005 On the 22nd, it was deposited at the General Microbiology Center (CGMCC) of the China Microbial Culture Collection Management Committee, and its deposit number is CGMCC NO.1446. Pick a single colony of HA9801 strain on a rabbit blood agar plate and inoculate it into Todd-Hewitt broth (THB) containing 5% calf serum (purchased from Weigang Serum Factory), incubate at 37°C overnight, and take 0.5 mL of bacterial solution Centrifuge, extract chromosomal DNA, dissolve it in PH8.0TE buffer, and freeze it for later use.

[0028] The method of extracting the genomic DNA of Streptococcus suis type 2 HA9801 strain is based on the method of M...

Embodiment 2

[0088] 2.1 Obtain the coding gene of Streptococcus suis type 2 GAPDH-MRP-HM6 fusion epitope

[0089] Streptococcus suis type 2 HA9801 strain Yao Huochun et al. (Journal of Nanjing Agricultural University, 1999, 22(2): 67~70) was isolated and identified from the diseased pigs with the outbreak of streptococcus suis in Jiangsu in 1998, and was identified in August 2005 On the 22nd, it was deposited at the General Microbiology Center (CGMCC) of the China Microbial Culture Collection Management Committee, and its deposit number is CGMCC NO.1446. Pick a single colony of HA9801 strain on a rabbit blood agar plate and inoculate it into Todd-Hewitt broth (THB) containing 5% calf serum (purchased from Weigang Serum Factory), incubate at 37°C overnight, and take 0.5 mL of bacterial solution Centrifuge, extract chromosomal DNA, dissolve it in PH8.0TE buffer, and freeze it for later use.

[0090] The method of extracting the genomic DNA of Streptococcus suis type 2 HA9801 strain is based on t...

Embodiment 3

[0153] Example 3 Expression of Streptococcus suis type 2 GAPDH-MRP-HM6 fusion gene

[0154] Transform the recombinant fusion plasmid into competent E. coli BL 21 (TaKaRa company product), screened with ampicillin plates. Pick a single colony to inoculate 5 mL of Amp-containing LB broth, shake at 28°C for 4 to 5 hours, inoculate 1000 mL of Amp-containing LB broth, and shake vigorously at 28°C for 4 to 5 hours. 600 When it reaches 0.6, add IPTG (product of Sigma) for induction, and shake vigorously at 37°C for 2 to 4 hours. Place the Erlenmeyer flask on ice for 5 minutes, centrifuge at 5000g at 4°C for 5 minutes, discard the supernatant, collect the bacteria, and use it immediately or freeze it.

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Abstract

The invention provides a vaccine composition and a preparation method thereof, belonging to the field of biological pharmacy. The vaccine composition comprises fusion protein formed by fusion expression of streptococcus suis type 2 virulence factor muramidase-released protein (MRP), glyceric aldehyde dehydrogenase (GAPDH) and newly-discovered membrane protein pyridine persulphuric acid nucleotide oxidation-reduction enzyme (HM6) gene, wherein the fusion protein has favorable immunogenicity and immunoprotective rate. The invention also provides a gene for coding the fusion protein as well as a reagent and a method for amplifying the gene.

Description

Technical field [0001] The invention relates to a genetically engineered recombinant protein vaccine, specifically, to a genetically engineered recombinant subunit vaccine for preventing swine streptococcus disease. Background technique [0002] Streptococcus suis (Streptococcus suis) is an important pathogenic bacteria of pigs. There are 35 serotypes currently known. Among them, serotype 2 (Streptococcus suis type 2, SS2) is a predominant strain in the world, which can cause pigs Acute sepsis, meningitis, arthritis, bronchial pneumonia and other diseases and acute death; at the same time, SS2 is an important pathogen of zoonotic diseases, and infects people, causing serious illness and even death. [0003] There is currently no genetically engineered vaccine for Streptococcus suis type 2 clinically. In this study, some immunogenic proteins of Streptococcus suis were analyzed, a certain immunogenic protein gene or multiple immunogenic protein genes were expressed in tandem, and th...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K19/00C12N15/62C12N15/63C12N1/15C12N1/19C12N1/21C12N5/10A61K39/116A61P31/04A61K39/09
Inventor 陆承平孙建和梁阳秋张炜范红结姚火春
Owner 陆承平