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| 正向和反向SV40 poly（A）信号序列对上游基因表达的影响 | |
| 引用本文： | 侯兵,金华君,刘文超,钱其军,吕赛群,刘文超. 正向和反向SV40 poly（A）信号序列对上游基因表达的影响[J]. 第二军医大学学报, 2010, 31(6): 630-633. DOI: 10.3724/SP.J.1008.2010.00630 |
| 作者姓名： | 侯兵 金华君 刘文超 钱其军 吕赛群 刘文超 |
| 作者单位： | 1. 第四军医大学西京医院肿瘤科,西安,710032;第二军医大学长海医院预防保健科,上海,200433 2. 第二军医大学东方肝胆外科医院病毒基因治疗实验室,上海,200438 3. 第四军医大学西京医院肿瘤科,西安,710032 |
| 基金项目： | 国家高技术研究发展计划重点项目("863计划") |
| 摘 要： | 目的研究正向和反向SV40poly(A)信号在3种常用细胞株中对上游基因表达的调控作用,为基因表达研究中载体poly(A)信号的选择提供依据。方法将F-Luc与R-Luc两种荧光素酶基因插入同一质粒,构建带有双荧光素酶报告基因的载体Dual-Luc。在F-Luc基因后分别插入正向和反向互补的SV40poly(A)信号序列,得到2个带有不同SV40poly(A)信号的载体Dual-Luc2、Dual-Luc3。将其分别转染常用的3类细胞株293、L-02和HeLa细胞,应用双荧光素酶标仪和反转录聚合酶链反应(RT-PCR)法测定报告基因(F-Luc)的表达量,以R-Luc基因的表达量作对照。结果成功构建携带正向和反向互补SV40poly(A)序列的双荧光素酶载体Dual-Luc2、Dual-Luc3;双荧光素酶标仪检测表明,在293细胞中,Dual-Luc2、Dual-Luc3的F-Luc/R-Luc比值分别为3.25±0.43、3.03±0.14,差异无统计学意义(P>0.05);在L-02细胞中,Dual-Luc2、Dual-Luc3的F-Luc/R-Luc比值分别为6.16±0.39、3.83±0.39,... |
| 关 键 词： | SV40 poly(A)加尾信号序列 基因表达 双荧光素酶报告系统 |
| 收稿时间： | 2009-12-10 |
| 修稿时间： | 2010-05-14 |
Influence of two kinds of SV40 poly (A) signals on upstream gene expression |
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| HOU Bing,JIN Hua-jun,LIU Wen-chao,QIAN Qi-jun,L Sai-qun. Influence of two kinds of SV40 poly (A) signals on upstream gene expression[J]. Former Academic Journal of Second Military Medical University, 2010, 31(6): 630-633. DOI: 10.3724/SP.J.1008.2010.00630 | |
| Authors: | HOU Bing JIN Hua-jun LIU Wen-chao QIAN Qi-jun L Sai-qun |
| Affiliation: | HOU Bing,JIN Hua-jun,LIU Wen-chao,QIAN Qi-jun,L(U) Sai-qun |
| Abstract: | Objective To investigate the regulatory effects of two kinds of SV40 poly(A) signals on the upstream gene expression in three cell lines,so as to provide theoretical evidence for selection of poly(A) of vectors.Methods A dual luciferase reporter vector Dual-Luc was constructed,and two SV40 poly(A) signal sequences were inserted in the downstream of the R-Luc gene separately.Then the two diverse dual luciferase reporter gene vectors Dual-Luc2 and Dual-Luc3 were transfected into 293,L-02,and HeLa cells.The re... |
| Keywords: | SV40 poly(A) signal gene expression dual luciferase reporter system |
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