人NHE-1基因反义真核表达载体的构建及初步鉴定  

Construction and identification of Na^+-H^+ exchanger-1 gene antisense eukaryotic expression vector

作　　者：滕小春[1] 刘海峰[1] 陈刚[1] 杨孟华[1] 杨仕明[1] 

机构地区：[1]第三军医大学西南医院全军消化内科中心,重庆400038

出　　处：《第三军医大学学报》2004年第2期112-114,共3页Journal of Third Military Medical University

基　　金：第三军医大学科研基金资助项目 ( 2 0 0 2 )~~

摘　　要：目的　构建人Na+ H+ 交换蛋白 1(Na+ H+ exchanger 1,NHE 1)基因反义真核表达载体。方法　采用分子克隆技术 ,用EcoRⅠ和HindⅢ双酶从NHE 1cDNApEAP切下所需目的片段 ,然后将该片段反向插入真核表达载体pcDNA3 1( ) Zeo的多克隆位点 ,最后对产生重组子进行琼脂糖凝胶电泳鉴定。结果　经琼脂糖凝胶电泳鉴定 ,将目的片段成功的连接到pcDNA3 1( ) Zeo载体上。结论　成功地将NHE 1目的片段反向插入到pcDNA3 1( ) Zeo中 ,构建了人NHE 1反义表达真核载体hNHE 1。Objective To construct the antisense expression vector of human Na + H + exchanger 1 (NHE 1). Methods NHE 1 gene fragment, cleaved from cDNA pEAP with Eco RⅠ and Hin dⅢ, was reversely inserted into the polyclone site of plasmid eukaryotic expression vector pcDNA3.1( )/Zeo. Finally, the constructed recombinant was identified by agarose gel electrophoresis. Results Agarose gel electrophoresis confirmed that the target fragment was successfully bound to pcDNA3.1( )/Zeo. Conclusion The antisense eukaryotic expression vector of NHE 1, namely, hNHE 1, has been constructed successfully by means of reversely inserting the target fragment into pcDNA3 1( )/Zeo.

关 键 词：NHE-1基因 胃肿瘤 反义技术 重组载体 

分 类 号：Q781[生物学—分子生物学] Q782