【摘要】 目的在大肠杆菌中表达丙型肝炎病毒(HCV)核心蛋白基因片段。方法利用PCR技术,扩增出357 bp的HCV核心蛋白基因片段,双酶切后将其插入到原核表达载体pET-32a,构建重组表达质粒pET-32a/HCVc。转化到大肠杆菌BL21中,IPTG诱导表达。SDS-PAGE鉴定表达情况,Western blot鉴定表达产物的抗原性。结果经IPTG诱导后获得了目的蛋白的融合表达;SDS-PAGE电泳显示其在32 000处有一条融和表达条带;Western blot结果显示其具有HCV抗原特异性。结论HCV核心蛋白在大肠杆菌中获得高效表达,而且表达产物有良好的抗原性。更多还原

【Abstract】 Objective To express hepatitis C virus (HCV) core protein gene fragment in E.coli. Methods A fragment of HCV core gene sequence 357 bp in length was amplified by PCR, digested with EcoRⅠ+Hind Ⅲand inserted to the plasmid vector pET-32a to construct recombinant HCVc/pET-32a plasmid, which was transformed into E.coli BL-21 and induced by IPTG for its expression. The expressed proteins obtained were identified by SDS-PAGE and Western blotting. Results The core sequence of HCV was amplified and after IPTG induction, a fusion protein of 32 000 was resulted exhibiting specific reaction with HCV-positive serum and high antigenicity. Conclusion It is possible to efficiently express HCV core protein in E.coli.更多还原