Cloning human β-catenin gene promoter and analyzing its activity

Objective To clone a 1.8 kb fragment upstream of the βcatenin gene and assay its promoter activity.Methods A 1.8 kb fragment upstream of the βcatenin gene was amplified by PCR using human genomic DNA as a template.Its promoter activity was determined with dual-luciferase reporter assay after it had been cloned into a pGL3-basic vector and transfected into PC3 cells.Results The sequence of the 1.8 kb fragment proved to be correct by DNA sequencing.Dual-luciferase reporter assay(Ml/M2) was 11.71 at 48 h after PGL3-1.8 kb was co-transfected with pRL-TK into prostate cancer cell PC3 which was about 2.43-fold higher than that of pGL3-control co-transfection with pRL-TK,206.31 fold higher than that of pGL3-basic co-transfection with pRL-TK and 21.38 fold higher than that of pGL3-promoter cotransfection with pRL-TK.Conclusion The cloned 1.8 kb fragment upstream of the β-catenin gene presented strong promoter activity.