丙型肝炎病毒核心蛋白反式激活SV40病毒早期启动子/增强子的研究
The study of transactivating effect of HCV core protein on SV40 early promoter /enhancer
-
摘要: 目的: 探讨丙型肝炎病毒核心蛋白的反式激活作用。方法: 以重组质粒pcDNA3 core (含有HCV核心蛋白编码基因)转染HepG2细胞,从转录和翻译水平鉴定核心蛋白的瞬时表达;与报告质粒pSV lacZ共转染HepG2细胞,检测β半乳糖苷酶表达活性,酶的活性反映了表达的核心蛋白对SV40病毒早期启动子/增强子功能的影响。结果: 质粒pcDNA3 core在HepG2细胞瞬时表达核心蛋白,共转染实验中pcDNA3 core组β半乳糖苷酶的表达是空质粒对照的5 4倍。结论: HepG2细胞中表达的核心蛋白具有反式激活SV40早期启动子/增强子的特性,为进一步以差异显示逆转录聚合酶链反应技术克隆相关基因提供实验基础。Abstract: Objective: To investigate the transactivating effect of HCV core protein Methods: A recombinant plasmid containing the encoding gene of HCV core protein was transfected into HepG2 cells and cotransfected HepG2 cells with reporter plasmid pSV lacZ by lipofectamine plus reagents HCV core protein produced in transient expression system was detected at the transcription and translation levels The activity of β galactosidase was detected by a kit,which reflected the transactivating function of HCV core protein Results: The core mRNA and protein can be detected in HepG2 cells lysate which was transfected by the pcDNA3 core The activity of β galactosidase in HepG2 cells transfected by the pcDNA3 core was 5 4 times as higher as that of control plasmid Conclusion: It is suggested that the recombinant plasmid pcDNA3 core can be expressed in HepG2 cell,and it has transactivating effect on SV40.early promoter/enhancer.
下载: