人SREBP-1c启动子报告基因的构建和功能验证

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

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 blot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。

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 successfully constructed 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 through inhibition the SREBP-1c transcription.