Construction and identification of luciferase reporter gene vector containing hMLH1 promoter

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(-1 953/+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 simulation by E2.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,P0.001).Conclusions The hMLH1 promoter contains the regulatory sequence associated with E2.