关键词: CYP2C19, 基因多态性, 真核表达, 转染

Abstract: AIM: To construct the eukaryotic expression vector of CYP2C19^*1,^*2,^*3, respectively, then stably transfect HepG2 cells with them. METHODS: The full-length CYP2C19 wild-type cDNA fragment was amplified by PCR from the human liver cDNA library and mutagenesis build ^*2, ^*3 (mutant) cDNA in vitro, then those cDNA were inserted into eukaryotic expression vector pcDNA3.l(-). After identification of restriction digestion and PCR,the recombinant plasmid was transfected into HepG2 cells by lipofectamine. After screening culture by G4l8, a stably-transfected cell line was established, and the transcription and expression of the CYP2C19 gene SNPs were identified by RT-PCR, Western Blot assay. RESULTS: The eukaryotic expression vector spcDNA3.l-CYP2C19^*1, ^*2, ^*3 were successfully constructed and transfected stably into HepG2 cells, respectively. Stably-transfected cell lines were established and the CYP2C19 gene SNPs was expressed successfully. CONCLUSION: The establishment of the stably-transfected cell line and the expression of the target gene provide a solid experimental foundation for screening and prediction the metabolic of new drugs on the function of the CYP2C19 gene.

Key words: CYP2C19, Gene polymorphism, Eukaryotic expression vector, Transfection