人SREBP-1c启动子报告基因的构建和功能验证

基础医学与临床 ›› 2009, Vol. 29 ›› Issue (5): 495-498.

人SREBP-1c启动子报告基因的构建和功能验证

刘晓军孔星星王瑞邵迪乔爱军常永生方福德

中国医学科学院基础医学研究所北京协和医学院基础医学院中国医学科学院基础医学研究所北京协和医学院基础学院中国医学科学院基础医学研究所北京协和医学院基础学院

收稿日期:2009-02-27 修回日期:2009-03-19 出版日期:2009-05-25 发布日期:2009-05-25
通讯作者: 方福德

Construction of pGL3-Basic-SREBP-1c-promoter reporter gene vector and detection of its function

Xiao-jun LIU, Xing-xing KONG, Rui WANG, Di SHAO, Ai-jun QIAO, Yong-sheng CHANG, Fu-de FANG

IBMS, CAMS&PUMC IBMS, CAMS&PUMC IBMS, CAMS&PUMC

Received:2009-02-27 Revised:2009-03-19 Online:2009-05-25 Published:2009-05-25
Contact: Fu-de FANG

摘要/Abstract

摘要： 目的构建人SREBP-1c启动子的荧光素酶报告基因载体并进行功能的初步验证。方法提取人血基因组DNA，PCR扩增SREBP-1c的启动子，构建SREBP-1c启动子双荧光素酶的报告基因载体；用双荧光素酶活性检测其功能。Western blotting检测FoxO1对SREBP-1c蛋白的抑制作用。结果成功构建了pGL3-Basic-SREBP-1c-promoter的报告基因载体，共转染FoxO1明显抑制了人SREBP-1c启动子的活性，同时过表达FoxO1抑制了SREBP-1c的蛋白表达。结论 FoxO1通过抑制SREBP-1c的转录来抑制SREBP-1c的表达。

关键词: SREBP-1c, 启动子, FoxO1, 双荧光素酶报告基因分析

Abstract: Objective To construct human SREBP-1c-promoter reporter gene vector and to detect its function.　 Methods Human blood genome DNA was extracted and pGL3-Basic-SREBP-1c-promoter reporter gene vector was constructed. Furthermore, the function of SREBP-1c-promoter was confirmed by dual-luciferase reporter assay. Results　pGL3-Basic-SREBP-1c-promoter reporter gene vector was constructed successfully and the promoter activity was obviously repressed by co-transfection FoxO1. Overexpression FoxO1 inhibited the SREBP-1c protein expression. Conclusion　FoxO1 repressed the SREBP-1c protein expression by inhibition the SREBP-1c transcription.

Key words: SREBP-1c, Promoter, FoxO1, Dual-Luciferase Reporter Assay

刘晓军孔星星王瑞邵迪乔爱军常永生方福德. 人SREBP-1c启动子报告基因的构建和功能验证[J]. 基础医学与临床, 2009, 29(5): 495-498.

Xiao-jun LIU; Xing-xing KONG; Rui WANG; Di SHAO; Ai-jun QIAO; Yong-sheng CHANG; Fu-de FANG. Construction of pGL3-Basic-SREBP-1c-promoter reporter gene vector and detection of its function[J]. Basic & Clinical Medicine, 2009, 29(5): 495-498.

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