Journal of Chemical and Pharmaceutical Research (ISSN : 0975-7384)

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Original Articles: 2014 Vol: 6 Issue: 6

Molecular cloning, expression and metabolic activity assay of CYP3A46 gene


The present study is to study drug metabolic characteristics of CYP3A46 gene. Sus scrofa CYP3A46 gene from Bama miniature pig liver was obtained by PCR method and cloned in pCDNA3.1, the recombinant plasmid pCDNACYP3A46 was obtained. The recombinant plasmid and empty plasmid were transfected into HepG2 which designated as HepG2-CYP3A46 and HepG2-CDNA3.1 repectively. The stably expressing cell lines HepG2-CYP3A46 and HepG2-pcDNA3.1 were achieved by the selection of gentamycin (G418) ,and were incubated with the probe drug nifedipine (NF) in optimal conditions. After the cells were incubated for 60 minutes, the high performance liquid chromatography (HPLC) was utilized to detect the change of metabolites in cells. The results showed that the sequence of cloned Sus scrofa CYP3A46 gene was correct, and the stably expressing cell the cell lines transfected with pCDNA- CYP3A46 and pCDNA3.1 were established successfully. The activity analysis demonstrated that the cell line transfected with pCDNA-CYP3A46 had the significantly metabolic activities of nifedipine( P<0.001).

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