论文部分内容阅读
目的：将中国人丙型肝炎病毒（HCV）核心基因cDNA克隆到真核细胞表达载体pTM3中，并在HepG2细胞中表达HCV核心蛋白．方法：利用DNA重组技术,将HCV／C基因cDNA定向克隆到pTM3中,采用痘苗病毒／噬菌体T7多聚酶短暂表达体系,在Lipofectin介导下将重组质粒转染HepG2细胞。结果：重组质粒转化Top10F＇细菌,随机挑取10个Amp抗性克隆,经EcoRⅠ、PstⅠ酶切鉴定，得到2个pTM3-Q534克隆。HepG2转染细胞以间接免疫荧光法鉴定证实存在HCV核心蛋白的表达．结论：HCV／C基因cDNA在粘粒载体中已获得克隆和初步表达．
OBJECTIVE: To clone the core gene cDNA of hepatitis C virus (HCV) from Chinese to eukaryotic expression vector pTM3 and express HCV core protein in HepG2 cells. Methods: The cDNA of HCV / C gene was cloned into pTM3 by DNA recombination technique. The recombinant plasmid was transfected into HepG2 cells mediated by Lipofectin using transient vaccinia virus / phage T7 polymerase. Results: The recombinant plasmids were transformed into Top10F ’bacteria. Ten Amp - resistant clones were randomly picked and identified by EcoR Ⅰ and Pst Ⅰ digestion. Two pTM3 - Q534 clones were obtained. HepG2 transfected cells were identified by indirect immunofluorescence to confirm the presence of HCV core protein. Conclusion: The cDNA of HCV / C gene has been cloned and expressed in cosmid vector.