关键词: 错配修复基因, hMLH1, 启动子, 雌激素, 双萤光素酶报告基因

Abstract: Objective To construct luciferase reporter gene vector containing human MLH1 promoter and to assay the transcriptional activity of hMLH1 promoter induced by E2. Methods hMLH1 promoter（-1953/+53）were amplified from the genomic DNA of human by PCR and cloned into luciferase reporter gene vector, pGL3-Basic. The recombined vector was transfected into HEK293 cells, and the activity of the luciferase was determined after the cells were simulated by E2 or not. Results The results of restriction enzyme digestion and sequencing indicated that the recombinant vector pGL3-Promoter1-luc was successfully constructed. After transcription of pGL3-Promoter1-luc, the activity fold of the luciferase was 7.45±0.81 induced by E2, which is significantly higher than 3.28±0.19 without E2(n=3,P<0.001). Conclusion The hMLH1 promoter contains the regulatory sequence associated with E2.

Key words: 错配修复基因, hMLH1, 启动子, 雌激素, 双萤光素酶报告基因