猪轮状病毒VP6基因的原核表达载体构建和表达

Construction and Expression of Porcine Rotavirus VP6 Gene Prokaryotic Expression Plasmid

根据已报道的猪轮状病毒VP6基因序列设计了2对引物,利用pGEM-T-Easy-VP6重组质粒为模板,经PCR扩增获得了1 194 bp的产物。将扩增片段分别插入到pET5α和pGEX-4T-2构建表达载体,并分别在大肠杆菌BL21(DE3)Plyss和ER2566中表达。结果表明:成功构建了重组质粒,而且该片段在大肠杆菌中也成功表达,获得了大小为44 kD的蛋白。

Based on the published VP 6 gene sequence of porcine rotavirus in GenBank, two pairs of primers were designed and synthesized. Using the primers, a gene fragment of approximately 1194bp was amplified by PCR from pGEM-T-Easy-VP6 recombinant plasmid. The amplified fragment was inserted into vector pETSa and pGEX-4T-2 respectively, after induction with IPTG. The result showed that the recombinant plasmids were successfully constructed,and the fragment was expressed in Escherichia coli BL21 （DE3）Plyss and ER2566 successfully, the molecular weight of the protein being 44 kD approximately.