丙型肝炎病毒核心蛋白基因的克隆及其高效表达

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357bp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32000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。

Cloning and expression of hepatitis C core protein gene]

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 III and 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.