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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 a mammalian cell line that can stably express the core protein of hepatitis C virus in order to further study the antigenicity and other functions of the core protein of hepatitis C virus. Methods: The HCV core gene was amplified by PCR, subcloned into eukaryotic expression plasmid pcDNA3.1 (+), and then transfected into HepG2 hepatoma cell line by liposome. The cell lines were infected with HepG2 and the core protein of HepG2 was detected by indirect immunofluorescence. Results: The cloned gene was 594bp in length and confirmed by sequencing and recombinant plasmid digestion. The target gene was confirmed by indirect immunofluorescence staining in HepG2 cells transfected with the target gene. . Conclusion: The core protein of hepatitis C virus can be stably expressed in HepG2 cells.