SCN5A基因启动区+495bp～+584bp片段的克隆与功能分析  

Cloning and functional analysis of the oligonucletides +495bp～+584bp in SCN5A promoter region

作　　者：张振明[1] 李晖[1] 董钦[1] 刘宇[1] 李凤兰[1] 马宁[1] 李金波[1] 刘端阳[1] 舒畅[1] 逄淑超[1] 

机构地区：[1]哈尔滨医科大学生物化学与分子生物学教研室,黑龙江哈尔滨150081

出　　处：《哈尔滨医科大学学报》2007年第6期527-530,共4页Journal of Harbin Medical University

基　　金：黑龙江省自然科学基金资助项目(D2005-29);黑龙江省教育厅海外学人重点项目资助(1054QH014);哈尔滨医科大学医学基础学科青年科学基金(060022);黑龙江省卫生厅科技项目基金(2006-272)

摘　　要：目的为了研究大鼠心肌SCN5A基因启动区+495bp^+584bp区域内的顺式作用元件对转录调控的影响。方法应用聚合酶链反应扩增大鼠心肌SCN5A基因启动区+495bp^+584bp片段,与荧光素酶报告基因载体pGL3-promoter重组,以pGL3-promoter空载体为对照,瞬时转染人胚肾母细胞(HEK293)和小鼠胚胎心肌细胞(H9C2),检测荧光素酶活性。结果成功构建的pGL3-P1重组载体的荧光素酶活性与pGL3-promoter对照载体相比,在HEK293细胞无明显差别(P>0.05),在H9C2细胞荧光活性增高(22.5±5.4)倍(P<0.01)。结论大鼠心肌SCN5A基因启动区+495bp^+584bp区域存在对基因的转录调控活性具有增强作用的组织特异性顺式作用元件。Objective To study the effect of oligonucleotides ＋ 495bp - ＋ 584bp of SCN5A promoter on gene transcription regulation and identify the potential cis-acting element in this region. Methods The ＋ 495bp - ＋ 584bp of SCN5A promoter was amplified by polymerase chain reaction（ PCR） and recombinated into pGL3- promoter luciferase report gene vector and transient transfected HEK293 cell and H9C2 cell. The activity of the luciferase report gene was detected. Results The recombinant expression vector was confirmed by restriction analysis and sequensing. Analysis of luciferase.report gene identified that the activity of luciferase report gene of pGL3-promoter in HEK293 cells had not obvious difference compared to the unloaded pGL3-promoter luciferase report gene in HEK293 cells, but its activities increased proximately （22.5 ± 5.4） times higher than that of pGL3-Promoter in H9C2 cell. Conclusion There is a tissue specific cis-acting element that can enhance the activity of transcription in ＋ 495bp - ＋ 584bp region of rat SCNSA.

关 键 词：SCN5A基因 转录调控 顺式作用元件 荧光素酶报告基因 

分 类 号：Q784[生物学—分子生物学]