Construction of pGL3-basic-FABP4-promoter Reporter Gene Vector and Detection of Its Function

Objective To construct an FABP4 promoter recombined luciferase reporter gene vector.and to detect its function,thus providing a basis for the research on lipid metabolism related medicines.Methods Primers were designed based on human FABP4 gene promoters,and FABP4promoters from human genome DNA was replicated by PCR.Then FABP4 promoters were inserted into the pGL3-basic vector after double digestion by restriction enzyme KpnI and XhoI.Positive clones were identified by sequencing.The recombinant pGL3-basic-FABP4-promoter vector were transiently transfected into K293 cells.After 24 hours,the activity of luciferase was detected.Results A pGL3 luciferase reporter vector was constructed.The result of sequencing and double digesting of recombined plasmid were completely correct.The results of luciferase detection after transfection showed that the transcriptional activity of recombinant pGL3 plasmid was obviously increased compared with that of pGL3- basic plasmid(P0.001).Conclusion A pGL3 luciferase reporter vector containing human FABP4 promoter gene was constructed successfully,which provides a tool and basis for further study of FABP4 gene expression regulation.