HCV p7下调基因p7TP2启动子序列的克隆及转录活性检测  

Promoter sequence cloning and its transcription activation evaluation of p7TP2 down-regulated by HCV p7 protein

作　　者：袁菊[1] 陶明亮[1] 靳亚平[1] 成军[2] 洪源[1] 

机构地区：[1]西北农林科技大学动物医学院,陕西杨凌712100 [2]北京地坛医院传染病研究所,北京100011

出　　处：《西北农林科技大学学报（自然科学版）》2008年第5期39-42,47,共5页Journal of Northwest A&F University(Natural Science Edition)

基　　金：国家自然科学基金项目(C03011402C30070689)

摘　　要：【目的】克隆p7TP2基因的启动子区域,并检测HCV p7TP2启动子的转录活性。【方法】通过分析确定p7TP2基因翻译起始密码子ATG上游1 203 bp至下游147 bp的基因组DNA序列作为启动子,克隆启动子片段并构建载体pCAT3-p7TP2-p和pGLB-p7TP2-p,转染HepG2细胞,应用CAT表达系统和双荧光素酶系统检测启动子活性。【结果】克隆了预测具有启动子活性的片段,成功构建了pCAT3-p7TP2-p和pGLB-p7TP2-p载体,转染pCAT3-p7TP2-p的HepG2细胞,CAT表达活性是转染pCAT3-Basic细胞的5.98倍;转染pGLB-p7TP2-p的HepG2细胞,双荧光素酶表达活性是转染pGLB细胞的9.67倍。【结论】克隆的p7TP2启动子序列与预测的序列一致,并且具有启动子转录活性。[Objective] The study was to clone the promoter sequence of p7TP2 gene and to evaluate the transcription activation of promoter sequence. [Method] The promoter sequence of p7TP2 gene（-1 203- 147 bp） was amplified from HepG2 genome by PCR, the promoter sequence was cloned into pCAT3 reporter vector and luciferase reporter which were transfected into HepG2 cell lines to evaluate its transcription activation. [Result] The promoter sequence of p7TP2 gene was amplified and cloned into the pCAT3 and pGLB. The CAT reporter system showed that the transcription activation of pCAT3-p7TP2-p was 5.98 times higher than pCAT3-Basic;the luciferase reporter system showed that the transcription activation of pGLB-p7TP2-p was 9.67 times higher than pGI.B. [Conclusion] The cloning promoter sequence was the same as the predicted sequence. The CAT reporter system and luciferase reporter system all successfully showed that the predicted promoter sequence had high transcription activation.

关 键 词：丙型肝炎病毒HCV p7TP2 启动子 转录活性 

分 类 号：Q78[生物学—分子生物学]