α-银环蛇毒素融合基因表达载体的构建及原核表达  

Construction of fusion gene vector of α-BGT and its expression in E.coli BL21(DE3)

作　　者：胡延春[1] 张乃生[2] 欧阳红生[2] 柳增善[2] 邓俊良[1] 

机构地区：[1]四川农业大学动物医学院,四川雅安625014 [2]吉林大学畜牧兽医学院,吉林长春130062

出　　处：《畜牧与兽医》2008年第4期7-10,共4页Animal Husbandry & Veterinary Medicine

摘　　要：设计并人工合成2条引物,对作者构建的重组质粒pMD-α-BGT进行PCR扩增获得α-BGT基因。将目的基因连接至pUCm-T载体构建克隆质粒pUCm-α-BGT。克隆质粒经BamHⅠ、NotⅠ双酶切后连接至融合蛋白表达载体pGEX-4T-1中,成功构建融合基因表达载体pGEX-α-BGT,转化大肠杆菌BL21(DE3)进行IPTG诱导表达。表达产物经15%SDS-PAGE分析,在约34ku处可见明显的外源蛋白质表达带,与预计的分子量大小一致,其表达量约占细菌总蛋白的32.6%。Westernblotting和间接ELISA检测结果表明,α-BGT的融合表达蛋白与天然α-BGT标准品具有相似的抗原性。Using the recombinant plasmid pMD-α-BGT as template, alpha-bungarotoxin (α-BGT) gene was obtained by PCR amplification. The cloning plasmid pUCm-α-BGT was constructed by ligating α-BGT gene into pUCm-T vector. α-BGT gene fragment was recovered by BamHⅠand Not I restriction digestion of pUCm-α-BGT, and finally ligated into BamHⅠand Not I digested pGEX-4T-1, generating the fusion gene expression vector pGEX-α-BGT. The expression vector was transformed into Escherichia coli BL21 (DE3) and the recombinant bacte...

关 键 词：Α-银环蛇毒素 克隆表达 抗原性 

分 类 号：Q78[生物学—分子生物学]