携带人多药抗性基因逆转录病毒载体的构建及其在NIH3T3细胞中的表达  

Construction of Retroviral Vectors Containing Human Multidrug Resistance Gene(mdrl) and mdrl Gene Expression Mediated by the Vectors in NIH3T3 Cells

作　　者：周生[1] 毛宁[1] 吴英[1] 王宜强[1] 杜德林[1] 唐佩弦[1] 潘凌亚[1] 

机构地区：[1]军事医学科学院基础医学研究所

出　　处：《基础医学与临床》1997年第3期189-194,共6页Basic and Clinical Medicine

基　　金：国家自然科学基金

摘　　要：从ｐＨａｍｄｒｌ／Ａ质粒用ＸｈｏⅠ和ＳａｃⅡ将ｍｄｒｌ基因切下后克隆到ｐＢｌｕｅｓｃｒｉｐｔＳＫ质粒的相应酶切位点获得测序质粒ｐＢＳｍｄｒｌ，测定了ｍｄｒｌ基因的两端序列。用ＥｃｏⅠＣＲＩ和ＸｈｏⅠ从ｐＢ－Ｓｍｄｒｌ切下ｍｄｒｌ基因，定向克隆到逆转录病毒载体ｐＬＸＳＮ和ｐＭＳＣＶ２．１，得到携带ｍｄｒｌ基因的逆转录病毒载体ｐＬｍｄｒｌＳＮ和ｐＭＳＣＶｍｄｒｌ。用ＰＡ３１７病毒包装细胞包装后三种携带ｍｄｒｌ基因的病毒具有相似的滴度。用此三种病毒分别感染ＮＩＨ３Ｔ３细胞后，以ｐＨａｍｄｒｌ／Ａ载体ｍｄｒｌ基因产物表达量最高。提示Ｈａｒｖｅｙ肉瘤病毒的启动子可能具有较高的强度。mdrl gene was subcloned from pHamdrl/A to the XhoⅠ SacⅡ site of pBluescriptSK to produce pBSmdrl and sequenced partially. The pBSmdrl plasmid was digested by EcoⅠ CRⅠ and XhoⅠ,and mdrl gene was cloned into retroviral vectors pLXSN and pMSCVmdrl to produce pLmdrlSN and pMSCVmdrl.The three vectors showed similar virus titer when packaged in PA317 amphotropic packaging cells. The pHamdrl/A vector expressed largest amounts of mdrl gene products among the three vectors.

关 键 词：多药耐药基因 载体构建 逆转录病毒 NIH3T3细胞 

分 类 号：Q782[生物学—分子生物学] R730.5[医药卫生—肿瘤]