Pgenesil-1质粒载体介导的表达ERCC1shRNA的质粒构建和鉴定  

Construction and identification of Pgenesil-1-Mediated Plasmid expressing ERCC1 shRNA

作　　者：黄勇[1] 王绍海[1] 胡沙[1] 王泽华[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院妇产科,湖北武汉430022

出　　处：《中国妇幼保健》2008年第14期2000-2003,共4页Maternal and Child Health Care of China

摘　　要：目的:设计构建Pgenesil-1质粒载体介导的表达ERCC1shRNA的重组体质粒。方法:以ERCC1为靶基因,以Pgenesil-1质粒为载体,设计构建重组体,根据ERCC1基因cDNA序列,设计有小发卡结构的2条寡核苷酸序列,克隆到空载体Pgenesil-1中,转化DH5а菌株,提取质粒,进行酶切鉴定和测序分析。结果:经酶切鉴定筛出的重组体测序结果和目的序列相同,重组体载体构建成功。结论:成功构建了能表达ERCC1shRNA的质粒载体Pgenesil-1/ERCC11和Pgenesil-1/ERCC12,为下一步研究ERCC1基因在卵巢癌细胞株顺铂耐药中的作用奠定了实验基础。Objective： To construct the recombinant expressing ERCC1 small nuclear RNA （shRNA） with Pgenesil - 1 - Mediated Plasmid. Methods： Three pairs of oligonucleotide sequences with hairpin conformation were designed, and then inserted into Pgenesil - 1 - Mediated Plasmid. After screened from transformed DHSastrains, Pgenesil - 1 - Mediated Plasmid Expressing ERCC1 shRNA, the siRNA eukaryotic expression vectors, were successfully identified by sequencing. Results： Three recombinant vectors were constructed success- fully. Conclusion： Construction of plasmids of Pgenesil - 1/ERCC11 and Pgenesil - 1/ERCC12 expressing ERCC1 shRNA will be helpful for further elucidation of mechanism in ovary cancer cell llne resisting platinum.

关 键 词：Pgenesil-1载体 ERCC1shRNA质粒 

分 类 号：R737.31[医药卫生—肿瘤]