Cloning and expression of cytochrome P450 2C9: an enzyme for drug metabolism / Nur Aziah Mohd Rasali @ Razali

Introduction: CYP2C9 is one of the major enzymes present in the liver. Many of the drugs available in the market such as warfarin, phenytoin and NS AIDS go trough this enzymes for metabolism. Thus there is a need to develop an in vitro enzymes system that study substrates or chemical compounds which alter CYP2C9 enzyme activity.
Method: The pCW2C9 and pACYC-OxR was cloned inside competent cells of Escherichia coli at 25°C, 3(EC and 37°C. The lengths of incubation time were 24 hours. 48 hours and 72 hours. The proteins expressed were confirmed and evaluated through immunoblotting.
Result: The CYP2C9 and OxR protein was successfully expressed by recombinant E.coli through biotechnology technique. The immunoblotting showed the correct protein of size were 35 kDa and 80 kDa.
Conclusion: The bacteria system in recombinant technology can be used to further investigate the type of traditional and modern drugs and food that are substrates or inhibitors of CYP2C9 and thus offer drug- herbs and drug- drug interaction study.