p53RFP基因启动子荧光素酶报告基因载体的构建及鉴定  

Construction and identification of luciferase reporter plasmid containing p53RFP gene promoter

作　　者：裴静娴[1] 王月刚[2] 张艺军[3] 刘城[2] 吴平生[2] 

机构地区：[1]广州医科大学附属第二医院心内科广州心血管疾病研究所,广东广州510260 [2]南方医科大学南方医院心内科,广东广州510515 [3]广州军区广州总医院干部病房二科,广东广州510010

出　　处：《广东医学》2017年第2期170-172,共3页Guangdong Medical Journal

基　　金：国家自然科学基金青年基金资助项目(编号:30800462);广州医科大学博士启动项目(编号:2012C45)

摘　　要：目的构建人p53RFP基因启动子序列不同截短片段的荧光素酶报告基因载体,研究启动子的转录活性。方法 PCR扩增人p53RFP基因启动子区域不同长度的目的片段,克隆到pGL3-Basic中,构建启动子区域3个不同截短片段的荧光素酶报告基因载体,经双酶切和基因测序鉴定正确后,转染HEK293细胞,双荧光素酶报告基因检测系统检测荧光素酶活性。结果成功构建了p53RFP启动子不同截短片段的荧光素酶报告基因载体,经双荧光素酶报告基因检测分析,3个启动子片段均具有转录活性。结论成功构建了人p53RFP基因启动子荧光素酶报告基因载体,为研究p53RFP基因的转录调控机制提供了实验基础。Objective To construct the luciferase reporter gene vector containing human p53RFP gene promoter,and study the transcriptional activity of the promoter.Methods The different fragments of p53RFP promoter were generated by PCR,and subsequently cloned into pGL3-Basic to construct luciferase reporter vectors of p53RFP gene promoter.The resulting vectors were verified by double enzyme digestion and DNA sequencing.The luciferase activities were detected by dual luciferase reporter system in HEK293 cells.Results Three luciferase reporter gene vectors containing p53RFP gene promoter were successfully constructed.The dual luciferase reporter assay showed that all the three vectors had promoter activity.Conclusion The luciferase reporter gene vectors containing p53RFP promoter region are successfully constructed,which provides a basis for study on transcriptional regulatory mechanisms of p53RFP gene.

关 键 词：p53RFP 启动子 荧光素酶报告基因 

分 类 号：R541.4[医药卫生—心血管疾病]