The construction of hepatitis B virus promoter-luciferase reporter gene system

Objective To analyze the promoter of HBV based on the gene sequence and construct the luciferase report gene vectors with S1,S2,ENⅠ/X and ENⅡ/C promoter fragments.Methods The pHBV 1.3 plasmid was used as template for target fragment amplification,and the acquired fragments were constructed with pGL3-Enhancer respectively.The luciferase activities were detected after transient transfection of the 4 constructs to Huh7 cell line respectively.Results The target fragments with the length of 121 bp,368 bp,437 bp and 237 bp were obtained and 4 report gene vectors with S1,S2,ENⅠ/X and ENⅡ/C promoter fragments were constructed successfully.Conclusion These results may make an important basis for the further study of HBV promoter activity and regulation of gene expression,as well as the development of new anti-HBV drugs.