摘要
聚合酶链反应 (PCR)扩增HBVX基因 ,克隆至真核表达载体 pVR10 12中 ,构建HBVX基因重组表达载体 pVR10 12 X ;以该质粒转染HepG2细胞 ,酶联免疫吸附法 (ELISA)检测细胞X蛋白的瞬时表达 ;与报告质粒pSV lacZ共转染HepG2细胞 ,用试剂盒法检测 β 半乳糖苷酶表达活性。结果质粒 pVR10 12 X在HepG2细胞瞬时表达X蛋白 ,共转染实验中 pVR10 12 X组 β 半乳糖苷酶的表达是空质粒对照的 3 2倍。表明构建的表达载体能在哺乳动物细胞中表达 。
Polymerase chain reaction was employed to amplify the HBV X gene from plasmid pCP10, and the product was cloned into pVR1012, then transfected HepG2 cells and cotransfected HepG2 cells with reporter plasmid pSV lacZ HBx protein produced by HepG2 cells was measured by ELISA method The activity of β galactosidase was measured by a kit, which reflected the transactivating function of HBx protein The results showed that HepG2 cells transfected by pVR1012 X could express HBx protein The expression of β galactosidase in HepG2 cells transfected by the pVR1012 X was 3 2 fold higher as that of control plasmid It is suggested that the recombinant plasmid pVR1012 X can be expressed in mammalian cell line, and has transactivating effect on SV40 early promoter
出处
《解放军医学杂志》
CAS
CSCD
北大核心
2001年第6期404-406,共3页
Medical Journal of Chinese People's Liberation Army
基金
军队回国留学人员启动基金资助课题! (编号 98H0 38)
