丙型肝炎病毒核心区蛋白在HepG2细胞中的表达

EXPRESSION OF HCV CORE PROTEIN IN HepG2 CELLS

目的 :构建能稳定表达丙型肝炎病毒核心区蛋白的哺乳动物细胞系 ,以便对丙型肝炎病毒核心区蛋白的抗原性及其他功能作进一步研究。方法 :通过PCR扩增丙型肝炎病毒核心基因 ,将其亚克隆入真核表达质粒pcDNA3.1(+)中 ,用脂质体包裹后转染肝癌细胞系HepG2 ,通过G4 18筛选获得稳定转染细胞系 ,并通过间接免疫荧光法检测HepG2中丙型肝炎病毒核心区蛋白的表达。结果 :克隆的基因全长 5 94bp ,并经测序和重组质粒双酶切证实为所需目的基因 ;经间接免疫荧光验证实转染了目的基因的HepG2细胞中有丙型肝炎核心区蛋白的表达。结论 :丙型肝炎病毒核心区蛋白能在HepG2细胞中稳定表达。

Objective:To construct the mammalian cell line which can express HCV core protein stably,for further studying its function in causing diseases by HCV.Methods:The gene of HCV core region was amplified by PCR,then subcloned into eukaryotic expression plasmid pcDNA3.1(+).The recombinant plasmid was tranfected into HepG2 cells with liposome.The objective protein was detected by indirect immunofluorescene assay.Result:The eukaryotic expression plasmid pcDNA3.1(+) containing HCV core gene was constructed successfully,then identified by sequencing,double enzyme digestion and PCR.The cells stably transfected the recombinant plasmid were achieved by G418 selecting.The protein of HCV core region was identified expressing in cytoplasma and memberane of HepG2 by IF assay.Conclusion:The protein of HCV core region can express in HepG2 stably.