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
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[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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