鸭坦布苏病毒囊膜E蛋白的原核表达
Prokaryotic Expression of E Protein from Duck Tembusu Virus
-
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。Abstract: The entire E gene from the duck Tembusu virus (strain WR) was amplified by RT-PCR, and cloned into the pEASY-T3 vector.The recombinant plasmids carrying the target fragment were digested with Bam HI and Xho I, and cloned into the pET-32 avector to construct recombinant plasmid to be named pET-E.pET-E was subsequently transformed into Escherichia coli BL21 (DE3) .The fusion protein was expressed by IPTG induction, and presented mainly as inclusion bodies.The positive western blotting result suggested a strong reactinogenicity of the protein.