猪水泡病病毒VP2基因抗原区在大肠杆菌中的表达

Expressing Antigen Region of VP2 Gene of Swine Vesicular Disease Virus in Escherichia coli

利用RT_PCR技术 ,以SVDVHK’70为材料 ,扩增出VP2基因抗原区。将目的基因的PCR扩增产物直接进行双酶切 ,然后将酶切产物与酶切后的表达载体pGEX 4T_1进行连接 ,转化BL2 1菌体并提质粒 ,经酶切、PCR鉴定为阳性的重组质粒命名为pGEX_VP2 ,并测序。测序结果表明 ,目的基因插入的位置、大小和读码框均正确 ,表达载体构建成功。将含有阳性质粒的BL2 1菌液经IPTG诱导后进行SDS_PAGE分析 ,出现预期的目的蛋白条带 ,此目的蛋白经Westernblot检测确定其有免疫活性。

The antigen region of VP2 gene of swine vesicular disease was amplified by RT-PCR and nPCR, the amplified fragments were cloned into the expression vector pGEX 4T-1.The insert position,the size and the reading frame were identified by PCR,restriction digestion and the sequence analysis of the recombinant plasmids. Results of SDS-PAGE indicated that the recombinant plasmids could express the VP2 gene of Swine vesicular disease. Western blot indicated that the positive serum of SVDV could react with expressed protein.