Construction of luciferase reporter gene vectors containing B7-H1 promoter region and promoter activity assay

Objective To construct luciferase reporter gene vectors containing B7-H1 promoter region and to detect promoter activity in Hep G2 cells. Methods Fragments of the B7-H1 gene 5'-UTR promoters were amplified by PCR and cloned into PGL3 basic vector and named as P88,P175,P203,P398,P723,and P960 respectively.Then Hep G2 cells,were transfected with these constructed plasmids. The expression of luciferase before and after the addition of IFN-γ was detected by dual-luciferase(LUC) reporter assay system kit. Results Five luciferase reporter gene vectors containing B7-H1 promoter region were successfully constructed. The recombinant constructs were transiently transfected into Hep G2 cells. After the addition of IFN-γ,LUC detection assay showed that the expression of LUC was significantly enhanced by P175,P203,P398,P723 or P960,but not by P88. Conclusions Luciferase reporter gene vectors containing B7-H1 promoter region were successfully constructed,which may be valuable for further study on B7-H1 promoter activity and molecular mechanism.