CYP2B7cDNA的克隆,鉴定及真核细胞重组表达载体的构建  

CLONING AND IDENTIFICATION OF HUMAN CYTOCHROMECYP2B7 GENE cDNA AND CONSTRUCTION OF ITS MAN-MALIAN CELLE XPRESSION RECOMBINANT

作　　者：吴健敏[1] 董海涛[1] 余应年[1] 朱丽君[1] 

机构地区：[1]浙江杭州医科大学病理生理学教研室及医学分子生物学实验室

出　　处：《癌变．畸变．突变》1995年第1期1-6,共6页Carcinogenesis,Teratogenesis & Mutagenesis

摘　　要：应用ＲＴ—ＰＣＫ技术和ＤＮＡ重组技术从人肝组织中克隆细胞色素Ｐ４５０２Ｂ７（ＣＹＰ２Ｂ７）基因的ＣＤ－ＮＡ片段至ｐＧＥＭ—３Ｚ载体上，经限制性内切酶切图谱分析和ＤＮＡ序列分析确证克隆的片段为ＣＹＰ２Ｂ７的ｃＤＮＡ。然后将该片段亚克隆入一合适真核细胞表达载体的ＢａｍＨＩ位点处，获得４个克隆。其中１个含有正向插入片段，３个含有反向插入片段。为导入哺乳类细胞中，建立稳定表达ＣＹＰ２Ｂ７ｃＤＮＡ的细胞株打下基础。With reverse transcription polymerase chain reaction(RT PCR)and DNA re-combinant technique ,a full length cDNA encoding the monooxygenase cytochrome P450 ⅡB7 isolated from human liver was cloned into a plasmid pGEM 3Z. The cDNA segment wasanalysed by its restriction map and DNA sequencing,and showed nothing other than CYP287cDNA. Then the CYP2B7 cDNA was subcloned into a mammalian episomal expression vec-tor. Four clones with CYP2B7 cDNA insert were identified,one in right ortentation and threein reverse orientation.

关 键 词：细胞色素P450 基因 基因克隆 CDNA 基因表达 

分 类 号：Q559.9[生物学—生物化学]