Chain coccus recombination subunit vaccine and prepn. thereof

A technology of Streptococcus suis and Streptococcus equi, which is applied in the field of genetically engineered recombinant protein vaccines, can solve the problems of poor immunogenicity and achieve the effect of convenient use and good immunogenicity

Inactive Publication Date: 2006-09-20
范红结 +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the M-like protein and MRP subunit vaccines expressed separately have relatively poor immunogenicity due to their small molecular wei

Method used

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  • Chain coccus recombination subunit vaccine and prepn. thereof

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Comparison scheme
Effect test

Embodiment 1

[0026] Embodiment 1: Obtain the coding gene of the M protein antigen epitope of Streptococcus equi subspecies zooepidemicus

[0027] Streptococcus equi subspecies zooepidemic ATCC35246, isolated from Sichuan, my country in 1977, is collected by ATCC. Pick the ATCC35246 strain single colony on the rabbit blood agar plate and inoculate it into Todd-Hewittbroth (THB) (purchased from Oxford Company) broth containing 5% calf serum (product of Weigang Serum Factory), and cultivate overnight at 37°C. Take 0.5mL bacterial liquid and centrifuge to extract chromosomal DNA, dissolve in PH8.0TE buffer, and freeze for later use.

[0028] The method for extracting Streptococcus equi subspecies zooepidemicus genomic DNA is with reference to the method according to Michael W Lema et al. Adding improvements, the steps are as follows: pick ATCC35246 colonies on the rabbit blood plate and inoculate them into Todd-Hewit broth, and culture overnight at 37°C; take 0.5mL of bacterial solution and c...

Embodiment 2

[0045] Acquisition of the Coding Gene of Streptococcus suis Type 2 MRP Antigen Epitope

[0046] 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 sick pig in Jiangsu Province in 1998, and was identified in August 2005. On the 22nd, it was deposited in the General Microorganism Center (CGMCC) of China Committee for Culture Collection of Microorganisms, and its preservation number is CGMCC NO.1446. Pick a single colony of the HA9801 strain on the rabbit blood agar plate and inoculate it into Todd-Hewitt broth (THB) broth containing 5% calf serum (purchased from Weigang Serum Factory), culture it overnight at 37°C, and take 0.5 mL of the bacterial liquid Centrifuge to extract chromosomal DNA, dissolve in pH 8.0 TE buffer, and freeze for future use.

[0047] The method for extracting Streptococcus equi subspecies zooepidemicus genomic DNA is with reference to the method a...

Embodiment 3

[0064] Construction of fusion of mrp gene fragment of Streptococcus suis type 2 with M gene fragment of Streptococcus equi subspecies zooepidemicus

[0065] Genes and clones

[0066] The amplified M-like protein gene fragment and the mrp gene fragment were purified respectively with a DNA recovery kit (product of Roche Company), and the specific steps were carried out according to the instructions. After purification, the M-like gene fragment was digested with BamHI and SacI (products of New England Company); the mrp gene fragment was digested with SacI and HindIII (products of New England Company). The digested product was recovered again with a DNA recovery kit.

[0067] The recovered M-like gene fragment and the empty pET-32a (+) plasmid (product of TaKaRa Company) double digested with BamHI and SacI were separated by T 4 Ligase (product of New England Company) was used for ligation, and the competent DH5α was transformed to select the recombinant bacteria on the ampicill...

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Abstract

A streptococci recombined subunit vaccine for preventing the streptococcicosis of pig includes the fusion protein of the horseí»s Streptococcus equi subsp zooepidemicus type M protein fragment and the pigí»s Streptococussuis 2 type lysozyme releasing protein fragment. Said fusion protein has high immunogenicity. The gene for coding said fusion protein is also disclosed.

Description

technical field [0001] The invention relates to a genetic engineering recombinant protein vaccine, in particular to a genetic engineering recombinant subunit vaccine for preventing porcine streptococcosis. Background technique [0002] Streptococcus suis is an important bacterial infectious disease that seriously endangers the world's pig industry and can cause human infection. The main pathogens of the disease are Streptococcus equi subsp zooepidemicus and Streptococcus suis, type 2. At present, antibiotics and chemical drugs are mainly used to prevent and control the disease. However, due to the shortcomings of drug resistance and drug residues, it is urgent to develop new vaccines. [0003] M-like protein (M-Like Protein) is a filamentous surface protein on the cell wall surface of Streptococcus equi subsp zooepidemicus, which is an important virulence factor and protective antigen of the bacterium (Timoney J F, Walker J, Zhou M, et al. Cloning and sequence analysis of ...

Claims

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Application Information

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IPC IPC(8): A61K39/09C07K19/00C12N15/62A61P31/04C12N1/20
Inventor 范红结陆承平姚火春
Owner 范红结
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