Cloning of cytochrome P-450 2C9 cDNA from human liver and its expression in CHL cells.

ABSTRACT:

Aim

Using bacterial, yeast, or mammalian cell expressing a human drug metabolism enzyme would seem good way to study drug metabolism-related problems. Human cytochrome P-450 2C9(CYP2C9) is a polymorphic enzyme responsible for the metabolism of a large number of clinically important drugs. It ranks among the most important drug metabolizing enzymes in humans. In order to provide a sufficient amount of the enzyme for drug metabolic research, the CYP2C9 cDNA was cloned and expressed stably in CHL cells.

Methods

After extraction of total RNA from human liver tissue, the human CYP2C9 cDNA was amplified with reverse transcription-polymerase chain reaction (RT-PCR), and cloned into cloning vector pGEM-T. The cDNA fragment was identified by DNA sequencing and subcloned into a mammal

SUBMITTER: Zhu GJ 

PROVIDER: S-EPMC4658375 | biostudies-literature | 2002 Apr

REPOSITORIES: biostudies-literature