多药耐药基因启动子高表达载体的构建与鉴定  

Construction and identification of the eukaryotic expressive vector pcDNA3-MDR1 promoter

作　　者：王向玲[1] 马道新[2] 赵建强[3] 纪春岩[2] 

机构地区：[1]济南铁路中心医院 [2]山东大学齐鲁医院 [3]济南市中心医院

出　　处：《山东医药》2005年第13期17-18,共2页Shandong Medical Journal

基　　金：山东省科技厅资助项目(No.031050104)

摘　　要：目的克隆多药耐药基因(MDR1)启动子,构建并鉴定真核表达载体pcDNA3-MDR1启动子。方法采用PCR方法从白血病多药耐药K562-AO2细胞中扩增MDR1启动子,克隆入T载体,酶切后与pcDNA3连接,电泳及DNA测序进行鉴定。结果PCR克隆出MDR1启动子,成功克隆入T载体,DNA测序证实序列正确,通过连接成功构建含正确目的基因的表达载体pcDNA3-MDR1启动子。结论成功构建了MDR1启动子高表达载体,为靶向治疗耐药肿瘤构建载体提供了实验基础。Objective To clone MDR1 gene promoter and construct the eukaryotic expressive vector pcDNA3-MDR1 promoter.Methods The MDR1 promoter fragment was amplified using PCR method from K562-AO2 cells and cloned into T vector,and linked with pcDNA3 plasmid after enzymation.It was tested by the methods of electrophoresis and DNA sequencing.Results The MDR1 promoter was cloned and we confirmed that the eukaryotic expressive vector containing MDR1 gene promoter was successfully constructed through PCR products electrophoresis and DNA sequencing.Conclusion The successful construction of expressive vectors containing MDR1 promoter provides an experimental basis for targeted gene therapy to the multidrug resistant tumor cells.

关 键 词：多药耐药基因 启动子 真核表达载体 构建方法 鉴定方法 白血病 

分 类 号：R733.7[医药卫生—肿瘤]