Cloning Of HTERT Core Promoter And Its Significance In Gene Therapy Of Laryngeal Carcinoma

AIM: 1. To clone the hTERT (human telomerase reverse transcriptase) core promoter and construct a luciferase expression vector driven by hTERT core promoter and identify the transcriptional activity of the vector in tumor cells and normal cells. 2. HSV-tk gene was placed downstream of hTERT core promoter and whether specific expression of tk gene is in tumor cells was detected. METHODS: hTERT gene core promoter was amplified by PCR using the total genomic DNA from HeLa cells as template. The amplified gene fragment was subsequently cloned into PGL3-basic vector. Then the expression vector was confirmed by restriction enzyme digestion and PCR analysis. The luciferase activity driven by the hTERT gene core promoter was identified after transient transfection of the expression vector into tumor cells and normal cells. HSV-tk gene was amplified by PCR using plasmid pBluescript-tk which...