MDR1基因shRNA真核表达质粒的构建  

Construction of shRNA eukaryotic expression plasmid of MDR1 gene

作　　者：李旭艳[1] 邵淑丽[1] 恽冬泽[1] 

机构地区：[1]齐齐哈尔大学,黑龙江齐齐哈尔161006

出　　处：《黑龙江畜牧兽医》2014年第2期38-40,206,共4页Heilongjiang Animal Science And veterinary Medicine

基　　金：黑龙江省自然科学基金项目(C200624);黑龙江省教育厅科学技术项目(11511447;12511611)

摘　　要：为了构建靶向MDR1基因的短发夹RNA(shRNA)真核表达重组质粒,试验针对MDR1基因序列选取3个干涉靶点,设计合成3条编码shRNA的DNA模板,分别定向克隆到真核表达质粒pSilencer 3.1-H1中构建重组质粒,再进行菌落PCR扩增和测序鉴定。结果表明:菌落PCR扩增得到与预期大小相同的阳性带,经DNA测序证实插入片段完全符合设计要求,序列无突变。说明成功构建了3个靶向MDR1基因的真核表达重组质粒pSilencer 3.1-H1 shRNA。To construct short hairpin RNA （shRNA） eukaryotic expression recombinant plasmid targeting MDR1 gene, three intervention targets were selected from the sequences corresponding to MDR1 gene, and were used to design and synthesize three DNA templates encoding shRNA. The shRNA templates were directionally cloned into the eukaryotic expression plasmid pSilencer 3.1 - H1 to construct the recombinant plasmid, respectively, and the shRNA recombinant plasmid pSilencer 3.1 - H1 were identified by colony - PCR amplification and DNA sequencing. The results showed that the positive bands same as the expected sizes were obtained by colony - PCR amplification. It was proved that all the inserted fragments were fully consistent with the design requirements, and there were no mutations in the sequences. The results indicate that the three the eukaryotic expression recombinant plasmids pSilencer 3.1 - H1 shRNA targeting MDR1 gene are successfully constructed.

关 键 词：MDR1基因 短发夹RNA(shRNA) 真核表达重组质粒 H1启动子 

分 类 号：S813.3[农业科学—畜牧学]