大鼠SCN5A基因内含子1 +204 bp^+757 bp转录调控功能分析  

Functional Analysis of Transcriptional Regulation of Intron 1 +204 bp-+785 bp in Rat SCN5A Gene

作　　者：刘宇[1] 李晖[1] 董钦[1] 张振明[1] 李凤兰[1] 李金波[1] 马宁[1] 

机构地区：[1]哈尔滨医科大学基础医学院生物化学与分子生物学教研室,哈尔滨市150081

出　　处：《医学分子生物学杂志》2007年第2期108-111,共4页Journal of Medical Molecular Biology

基　　金：黑龙江省自然科学基金(NoD2005-29);哈尔滨医科大学研究生创新基金~~

摘　　要：目的克隆大鼠心肌SCN5A基因5′端调控区,检测目的片段在HEK293细胞中的转录活性。方法扩增大鼠心肌SCN5A基因+204 bp^+757 bp、+204 bp^+385 bp和+204 bp^+329 bp片段,构建含目的片段的荧光素酶报告基因——pGL3-P0、pGL3-P1和pGL3-P2,比较HEK293细胞中荧光素酶活性,评价不同长度DNA片段的转录调控活性。结果成功构建大鼠心肌SCN5A基因5′端3个片断的荧光素酶报告基因,HEK293细胞中pGL3-P1相对荧光素酶活性分别为pGL3-P0、pGL3-P2的4.3倍和2.8倍。结论SCN5A基因内含子1目的片段在HEK293细胞中具有较强的转录调控活性;SCN5A基因+329 bp^+385 bp为正调控区,软件分析MZF1、USF、C-ETs-1等因子能与这些正调控区域结合并可能参与SCN5A基因的表达调控。Objective To clone 5＇-flanking regulation sequence of rat SCN5A gene, and to detect the transcriptional activity of the target fragments in HEK293. Method The fragments ＋ 204 bp - ＋785 bp, ＋204 bp - ＋385 bp and ＋204 bp - ＋329 bp of SCN5A gene were cloned and constructed into luciferase reporter gene. The activities of luciferase were compared and the transcriptional activities of the DNA fragments of various lengths were evaluated in HEK293. Results The luciferase reporter gene with three fragments in 5＇-flanking region of SCN5A gene was successfully constructed. The relative luciferase activity of pGL3- P1 in HEK293 was 4. 6 times and 3 times that of pGL3-P0 and pGL3-P2 respectively. Conclusion The target fragments of SCN5A intron 1 have strong activity of transcriptional regulation in HEK293, a positive regulation region exists between ＋ 329 bp and ＋ 385 bp in intron 1, and the region can be bound by MZF1, USF and C-ETs- 1 which might be involved in the positive gene regulation.

关 键 词：SCN5A基因 转录调控 报告基因分析 顺式作用元件 

分 类 号：Q784[生物学—分子生物学]