猪瘟病毒石门株NS3基因在大肠杆菌中的表达

Expression of NS3 gene of CSFV Shimen strain in E. coli

采用PCR技术,扩增了猪瘟病毒石门株NS3基因的丝氨酸蛋白酶、NTPase、RNA解螺旋酶活性功能区长度为2 000 bp的片段,将其克隆到pET-32a中,构建了原核表达载体pET-NS3,并将其在大肠杆菌Rosetta(DE3)中进行了表达,同时对表达产物进行了SDS-PAGE和Western blotting检测。结果表明,IPTG诱导表达得到的pET-NS3融合蛋白的相对分子质量约为97 ku,与预期结果一致,且该重组蛋白能被CSFV阳性血清所识别。

Serine protease and NTPase/RNA helicase functional domains, 2.0 kb of NS3 gene of CSFV Shimen strain were amplified by PCR,and cloned into prokaryotic expression vector pET-32a. The target protein was expressed in E. coli Rosetta （DE3） induced with IPTG,and detected by SDS-PAGE and Western blotting. The results indicated that the 97 ku expressed protein could be recognized by CSFV positive serum as expected.