HBV/HAV复合抗原基因在CHO细胞中的表达

Expression of a Fused Gene for the Multiple Epitopes Antigen of Hepatitis B Virus and Hepatitis A Virus in Chinese Hamster Ovary Cells

为探讨HBV/HAV复合疫苗的可行性,利用DNA重组技术将HBsAg与HAW复合多表位抗原基因VPX进行融合,插入到真核表达质粒pVAX1多克隆位点,构建成核酸表达疫苗pVAX1/SVPX,将其转染CHO细胞。转染细胞培养48h后,进行RT-PCR、Western-blot、ELISA分析,结果表明HBV/HAV复合抗原基因SVPX能够在CHO细胞内转录、表达,融合蛋白具有HBV和HAV抗原的特性。

In order to explore the possibility of developing a HBV/HAV combined gene vaccine,a multi-epitope antigen gene VPX of HAV was fused with HBsAg gene and SVPX gene was constructed.SVPX was sub-cloned into an eukaryotic expression plasmid pVAX1 to construct the eukaryotic recombinant expression plasmid pVAX1/SVPX.The identified recombinant was then transfected into CHO cells with liposome mediated method.The transfected cells was tested by RT-PCR,Western-blot,ELISA method 48 hours after culture.The results showed that the multi-eptitope gene SVPX can be transcripted and expressed,and expressed fused protein could react with HAV and HBV antisera.