Cloning of CYP2J3 gene and its expression in 293 cell

Objective To clone rat CYP2J3 gene and investigate the expression of pcDNA 3.1(+)-CYP2J3 eukaryotic expression vector in 293 cells.Methods The total RNA was extracted from rat hepatic tissue.CYP2J3 gene was amplified by RT-PCR and OE-PCR,and cloned into the pcDNA 3.1(+) to form an eukaryotic expression vector.293 cells was transfected with liposome/plasmid pcDNA 3.1(+)-CYP2J3.Western blotting was used to detect the expression of pcDNA 3.1(+)-CYP2J3 in 293 cells.Results CYP2J3 gene was obtained by RT-PCR and OE-PCR.The recombinant eukaryotic expression vector for CYP2J3 gene had been successfully constructed.The results of Western blotting showed the transfected 293 cells could express CYP2J3 protein.Conclusion Successful clone of CYP2J3 gene and construction of Pcdna3.1(+)-CYP2J3 provide the possibility for the further study of the biological function and related disease.