Abstract:

To clone a 1.8?kb fragment upstream of the βcatenin gene and assay its promoter activity． MethodsA 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． ResultsThe 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. ConclusionThe cloned 1.8?kb fragment upstream of the βcatenin gene presented  strong promoter activity．

Key words: βcatenin gene； Cloning； Promoter; PC3 cellβ