HCV包膜糖蛋白基因真核表达载体的构建及其在HepG2中的瞬时表达

Construction of Eukaryotic Expression Vector of HCV Envelope Glycoprotein Gene and Its Transient Expression in HepG2 Cells

通过PCR法扩增出HCV包膜糖蛋白E1-E2的全长基因片段。将测序正确的片段克隆入真核表达载体pcDNA3.1(-)中,构建HCV包膜糖蛋白基因真核表达载体。再采用脂质体法转染肝癌细胞系hepG2,利用RT-PCR、Western blot等方法对目的基因的转录与表达进行分析与鉴定。结果表明所构建的含有HCV包膜糖蛋白E1-E2基因的真核表达载体在HepG2细胞中能瞬时表达。重组真核质粒的构建及表达为下一步建立稳定转染细胞系及进一步研究HCV包膜糖蛋白的功能奠定了基础。

Full length gene fragment of HCV envelope glycoprotein was amplified by PCR.The target genes were identified by enzyme digestion and sequencing and then cloned into the eukaryotic expression vector pcDNA3.1（-）.The recombinant plasmid was transfected into hepatocarcinoma cell line HepG2 by lipidosome method.RT-PCR and Western blot were performed to determine the transcription of the genes.The results show that the recombinant plasmid is constructed successfully,and the gene is transient expressed in HepG2 cells.It is useful for construction stable transfected cell lines and further study of the function of HCV envelope glycoproteins.