Cloning of human Nkx3.1 gene promoter and assay of its promoter activity

Objective: To clone 1.06kb fragment upstream of Nkx3.1 gene and assay its promoter activity. Methods: 1.06kb fragment upstream of Nkx3.1 gene was amplified by PCR using human genomic DNA as template. Its promoter activity was determined with dual-luciferase reporter assay after it had been cloned into pGL3-basic vector and transfected into LNCaP cells. Results: The sequence of the 1.06kb fragment proved to be correct by DNA sequencing. Dual-luciferase reporter assay (M1/M2) was 2.7 at 48 hours after pGL3-1.06kb was cotransfected with pRL-TK into prostate cancer cell LNCaP, which was about 1.5-fold higher than that of pGL3-control cotransfection with pRL-TK and 50-fold higher than that of pGL3-basic cotransfection with pRL-TK. Conclusion: Cloned 1.06kb fragment upstream of Nkx3.1 gene presented a strong promoter activity. [