Construction of a Luciferase Reporter Gene Vector Directed by p27 Promoter and Characterization of the Promoter Activity

Objective:To clone p27 promoter and insert it into a luciferase reporter gene vector,and to characterize the promoter activity.Methods:The promoter of p27 was amplified from human breast cancer cell line MCF-7 genomic DNA by PCR and cloned into pGL3-basic.The resulting plasmid was determined by DNA sequencing.The promoter activity was analyzed in 293T cell line.Results:The result of DNA sequencing showed that the sequence of the cloned promoter was correct.The activity of the p27 luciferase reporter gene was confirmed,and this activity was increased by estrogen receptor(ER)α in a dose-dependent manner.Conclusion:The p27 promoter was cloned successfully,and this will be an important basis for the study of the functions of ERα coregulators.