人抗药基因的克隆  

Cloning of Full-length Multidrug Resistance Gene, in Human Normal Liver

作　　者：黄承诚 吴孟超[1] 陈汉[1] 屠振兴[1] 李岱宗[2] 徐国威[2] 严根宝[2] 顾健人[2] 

机构地区：[1]第二军医大学肝胆外科研究所,200433 [2]上海市肿瘤研究所,200032

出　　处：《第二军医大学学报》1992年第5期411-414,共4页Academic Journal of Second Military Medical University

摘　　要：本研究根据抗药基因(Multidrug resistance gene_1,MDR_1)的cDNA序列,设计了3对DNA引物,它们的起止位置分别为:Ⅰ-64～46(5’)和1680～1698(3’);Ⅱ1471～1489(5’)和2905～2923(3’);Ⅲ2729～2747(5’)和3845～3863(3’)。用PCR技术分别将这3对引物间的cDNA从人正常肝组织中的mRNA中克隆出,它们的长度分别为1762bp,1452bp和1134bp。前两者均分别克隆于pBluescript SK,后者克隆于pGEM7Z。所得3个克隆分别称为pMDR1.7,pMDR1.4和pMDR1.1。DNA序列分析证实这3个克隆均为MDR_1基因。进一步亚克隆得到MDR_1的全长基因,长度为3927bp(pMDR3.9)。MDR_1全长基因的克隆成功为肿瘤抗药性的分子诊断和基因治疗提供了最基本的手段。Overexpression of multidrug resistance gene, (MDR1) is responsible for the resistance of anticancer agents in cancer cells. In this study, we designed three pairs of DNA primers to clone MDR, cDNA from human normal liver by polymerase chain reaction. The sites of these primers in MDR cDNA are (I)-64-46 (5') and 1680-1698 (3'); (II) 1471-1489 (5') and 2905-2923 (3'); (III)2729-2747 (5') and 3845-3863 (3). The three reactions were underwent after the human normal liver mRNA was reverse transcripted into single strand DNA. The lengths of PCR products are 1762bp, 1452 bp and 1134bp, respectively. The former two products were subcloned in pBluscript SK, respectively and the latter product was subcloned in pGEM7Z (named as pMDR1.7, pMDR1.4 and pMDRl.l, respectively). Cloned genes were certificated as MDR1 cDNA by sequence analysis. Full-length MDR, cDNA was obtained after further subcloning. Full-length MDR1 cDNA we obtained will be a very important tool in molecular diagnosis and gene therapy of anticancer drug resistance.

关 键 词：抗药基因 聚合酶链反应 肝脏 人类 

分 类 号：R967[医药卫生—药理学]