Clone and Activity Identification of Human NDRG2 Promoter

Objective: To clone the human NDRG2 promoter and analyze the promoter activity. Methods: The human NDRG2 promoter was amplified from BAC clone R-998D10 with the Advantage-GC genomic polymerase. The resulting amplicon was cloned into the pGL3-basic vector to generate the NDRG2/luciferase reporter plasmid. Various mutants NDRG2(-1131/+274), NDRG2(-273/+274), NDRG2(-135/+274), NDRG2(-79/+274) and NDRG2(-79/+57) were generated with PCR using the cloned promoter as template. All of the newly constructed plasmids were confirmed by sequencing. Transfect the plasmids into HEK293 and HeLa cells to analyze the promoter activity and identify the core region of human NDRG2. Results: The promoter of human NDRG2 was amplified successfully and the basic activity was analyzed. The result of sequencing was accordant with the one reported. Identify the core promoter region of human NDRG2. Conclusion: The activity of human NDRG2 promoters weakened with the consecutive truncation. The core region of human NDRG2 located -79 to +57.