p53RFP基因启动子荧光素酶报告基因载体的构建及鉴定

发布时间：2018-03-31 01:12

本文选题：pRFP　切入点：启动子　出处：《广东医学》2017年02期

【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。
[Abstract]:Objective to construct a luciferase reporter gene vector with different truncated fragments of human p53RFP promoter sequence and to study the transcriptional activity of promoter. Methods the target fragments with different length in the promoter region of human p53RFP gene were amplified by PCR and cloned into pGL3-Basic. Three luciferase reporter gene vectors with different truncated fragments in promoter region were constructed and identified by double enzyme digestion and gene sequencing, and then transfected into HEK293 cells. The luciferase activity was detected by double luciferase reporter gene detection system. Results the luciferase reporter gene vector with different truncated fragments of p53RFP promoter was constructed successfully. Conclusion Human p53RFP gene promoter luciferase reporter gene vector was constructed successfully, which provides an experimental basis for studying the transcriptional regulation mechanism of p53RFP gene.
【作者单位】：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;南方医科大学南方医院心内科;广州军区广州总医院干部病房二科;
【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)
【分类号】：R3416

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