摘要
利用RT-PCR方法扩增鸭坦布苏病毒分离株(WR株)全长E蛋白基因,克隆到pEASY-T3载体中,经Bam HI和XhoI酶切后将目的片段连接入pET-32a载体,构建原核表达重组质粒pET-E。表达质粒转化入感受态细胞BL21(DE3)后,经IPTG诱导后表达出鸭坦布苏病毒E蛋白,并以包涵体的形式存在,Westernblotting试验呈阳性,表明E蛋白有很好的反应原性。
The entire E gene from the duck Tembusu virus （strain WR）was amplified by RT-PCR,and cloned into thepEASY-T3 vector.The recombinant plasmids carrying the target fragment were digested with Bam HI and Xho I,andcloned into the pET-32a vector to construct recombinant plasmid to be named pET-E.pET-E was subsequently transformedinto Escherichia coli BL21 （DE3 ）.The fusion protein was expressed by IPTG induction,and presented mainly asinclusion bodies.The positive western blotting result suggested a strong reactinogenicity of the protein.
出处
《福建农业学报》
CAS
2014年第10期935-938,共4页
Fujian Journal of Agricultural Sciences
基金
福建省科技计划项目--省属公益类科研院所基本科研专项(2011R1025-8)
现代农业产业技术体系建设专项(CARS-43)
福建省种业创新与产业化工程建设项目(2011FJZY-9)
关键词
鸭坦布苏病毒
E基因
原核表达
duck tembusu virus
E gene
prokaryotic expression
