CYP3A39基因的克隆、表达及活性研究  

Cloning,Expression and Activity Analysis of S.scrofa CYP3A39

作　　者：薛正楷[1,2] 叶彬[2] 魏泓[3] 

机构地区：[1]泸州职业技术学院生物医学工程研究所,四川泸州646005 [2]重庆医科大学分子医学与肿瘤中心,重庆400016 [3]第三军医大学实验动物中心,重庆400038

出　　处：《河南农业科学》2013年第12期124-128,共5页Journal of Henan Agricultural Sciences

基　　金：国家"十五"科技攻关计划重点项目资助课题(2004BA717B23)

摘　　要：为研究细胞色素氧化酶CYP3A39的生物学活性,采用RT-PCR方法从巴马小型猪肝组织中克隆CYP3A39基因,与真核表达载体pcDNA3.1连接,构建重组质粒,转染HepG2细胞,采用G418筛选,以获得稳定表达该基因的细胞株;以CYP3A特异性代谢底物睾酮为探针药物,在体外进行孵育,高效液相色谱仪测定睾酮羟基化产物6β-羟基睾酮(6β-OHT)的产量。结果显示,成功克隆了CYP3A39基因,建立了稳定表达CYP3A39基因的HepG2细胞株,且其具有睾酮代谢活性。The CYP3A39 gene was cloned from the liver tissues of Bama miniature pig by RT- PCR,ligated to the expression vector pcDNA3. 1, and then transfected into HepG2 cells. The HepG2- CYP3A39 cell line, stably expressing CYP3A39 was established under the selection of gentamycin （G418）. The probe drug testosterone （TST） specific to CYP3A was used to incubate with the cell lines of HepG2-pcDNA3.1 and HepG2-CYP3A39 in optimal conditions,and the high performance liquid chromatography （HPLC） was utilized to detect the metabolites after the end of incubation. The results Showed that the cell line expressing CYP3A39 was successfully estab- lished and the expressed CYP3A39 had significantly higher metabolic activity of testosterone than that of negative control.

关 键 词：细胞色素氧化酶 CYP3A39 基因克隆 重组细胞 代谢活性 

分 类 号：S859.7[农业科学—临床兽医学]