Streptococcus agalactiae BibA recombinant protein, as well as coding gene preparation method and application thereof

A technology of recombinant protein and Streptococcus lactis, which is applied in botany equipment and methods, microbial-based methods, biochemical equipment and methods, etc., can solve the problem of poor cross-protection, few protective antigens, and inactivation of Streptococcus agalactiae cells. There are differences in the immune protection effect of live vaccines, etc., to achieve the effect of good reactogenicity

Inactive Publication Date: 2014-07-09
LANZHOU INST OF ANIMAL SCI & VETERINARY PHARMA OF CAAS
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are many technical difficulties in the development of conventional Streptococcus agalactiae vaccines: 1) There are differences in the immune protection effect of inactivated Streptococcus agalactiae vaccines; 2) There are

Method used

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  • Streptococcus agalactiae BibA recombinant protein, as well as coding gene preparation method and application thereof
  • Streptococcus agalactiae BibA recombinant protein, as well as coding gene preparation method and application thereof
  • Streptococcus agalactiae BibA recombinant protein, as well as coding gene preparation method and application thereof

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Experimental program
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Embodiment 1

[0054] Reagent source:

[0055] T-Vector pMD20: TaKaRa Bao Biological Engineering (Dalian) Co., Ltd., product number 3270.

[0056] cold TM I DNA: TaKaRa Bao Biological Engineering (Dalian) Co., Ltd., product number 3361.

[0057] 1. Primer design:

[0058] Design a pair of primers, in order to facilitate the subcloning in the next step, the upstream and downstream 5' ends were respectively introduced with restriction enzyme sites;

[0059] The upstream primer is: 5'-CGC GGATCC TCGGATTCAATTCCTCAT-3' (underlined in italics Bam HI restriction site);

[0060] The downstream primer is: 5'-CG GAATTC TGCATAATATCCAGGTGTAGGC-3' (underlined in italics Eco RI restriction site), and the primers were synthesized by Beijing Liuhe Huada Gene Technology Co., Ltd.

[0061] 2. Amplification and acquisition of BibA gene:

[0062] The genomic DNA of Streptococcus agalactiae isolates was extracted with a bacterial genomic DNA extraction kit, and used as a template for PCR amplif...

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Abstract

The invention discloses streptococcus agalactiae BibA recombinant protein, as well as a coding gene, a preparation method and an application thereof. The streptococcus agalactiae BibA recombinant protein provided by the invention is a protein sequence as shown in sequence No. 2. The invention also provides the coding gene, the preparation method and application of the protein. The streptococcus agalactiae BibA recombinant protein has the beneficial effects that streptococcus agalactiae is used as a research object, streptococcus agalactiae DNA is cloned to obtain a BibA gene, the cloning and sequencing of the gene sequence are completed, the prokaryotic expression and immunizing protection of the streptococcus agalactiae are researched, a vaccine development candidate antigen with high specificity and immune protection force is obtained, and the foundation is laid for prevention of streptococcus agalactiae diseases. The streptococcus agalactiae BibA recombinant protein has the advantages that the streptococcus agalactiae BibA gene is obtained by cloning, BibA recombinant expression plasmids are constructed, the BibA recombinant protein is obtained, westernblot detection results show that the BibA recombinant protein has relatively good reactogenicity, and animal (rat) experiments prove that the recombinant protein has certain protectiveness.

Description

technical field [0001] The invention relates to Streptococcus agalactiae BibA recombinant protein and its encoding gene, preparation method and application in the field of bioengineering. Background technique [0002] Streptococcus agalactiae is a zoonotic pathogen that can infect humans, cows, rabbits, tilapia, bullfrogs and other vertebrates. In human medicine, Streptococcus agalactiae is called group B streptococcus (GBS). GBS can cause premature delivery, late miscarriage, fetal dysplasia, premature rupture of membranes, endometritis, urinary tract infection, etc. in pregnant women with bacteria; Streptococcus agalactiae can cause acute, subacute and chronic mastitis in dairy cows. [0003] At present, Streptococcus agalactiae can be divided into 10 serotypes. Clinical studies have shown that the drug resistance of this bacterium is increasing year by year, so vaccine prevention of this disease has become the primary goal of scientific researchers. However, there are ma...

Claims

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Application Information

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IPC IPC(8): C07K14/315C12N15/31C12N15/70C12N1/21A61K39/09A61P31/04C12R1/19
CPCC07K14/315
Inventor 王旭荣李建喜杨志强王学智常瑞祥王磊张景艳秦哲孔晓军孟嘉仁
Owner LANZHOU INST OF ANIMAL SCI & VETERINARY PHARMA OF CAAS
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