HCV核心蛋白在大肠杆菌中的表达及免疫学特性分析

Expression of Hepatitis C Virus Core Protein in E.coli and Its Immunological Characteristics

目的:表达HCV核心蛋白,为检测丙肝病毒提供合适抗原。方法:以含HCV核心全长cDNA克隆的pMD18T/core质粒为模板,PCR扩增全长的HCV核心抗原基因,插入表达载体pQEN1构建重组质粒pQEN1/Core,转化BL-21(DE3)大肠杆菌,IPTG诱导表达6×His融合蛋白,表达产物经SDS-PAGE及Western blot检测和鉴定。结果:经SDS-PAGE及Western blot显示HCV核心蛋白在大肠杆菌中正确表达,融合蛋白分子量约为22 kD,表达量约占菌体蛋白总量的30%。纯化后的C蛋白能与慢性丙型肝炎患者有血清反应。结论:HCV核心蛋白在大肠杆菌中成功表达并具有较强的抗原性。

Objective： To express the recombinant HCV core protein and lay a foundation for studying and developing of HCV diagnostic antigens. Methods： A 573 bp cDNA encoding 191 amino acid of HCV type-1 core protein was cloned into a expression plasmid pQEN1 to construct recombinant plasmid pQEN1-Core. The recombinant plasmid was introduced into E. coli BL21 （DE3）. The expressed fussion protein was analyzed by SDS-PAGE, Western-blot and indirect ELISA, respectively. Results： The expressed fussion protein was shown by a major band with a expected molecular weight about 22 kD on SDS-PAGE and accounted for almost 30 % of the total bacteria proteins, Serological assay by indirect ELISA showed that it reacted with patient sera strongly and specifically. Conclusion： The recombinant HCV core protein was correct expressed and showed high specificity and good antigenicity.