针对人蛋白激酶CK2α亚基的shRNA表达质粒的构建与鉴定  

Construction and identification of shRNA expression plasmid targeted against protein kinase CK2α

作　　者：朱汝森[1] 刘新光[1] 梁念慈[1] 

机构地区：[1]广东医学院生物化学与分子生物学教研室,广东湛江524023

出　　处：《广东医学院学报》2006年第5期451-453,458,共4页Journal of Guangdong Medical College

基　　金：广东省教育厅自然科学研究项目(Z03042);湛江市科技局2003年招标项目;广东省医学科研基金项目(A2004454)

摘　　要：目的:构建针对人蛋白激酶CK 2α亚基的shRNA表达质粒。方法:以mU 6pro为载体,化学合成shRNA表达模板,并定向克隆到载体的U 6启动子下游,琼脂糖凝胶电泳法筛选阳性克隆,基因测序法检测插入模板序列的正确性。结果:获得了正确的重组质粒,插入的转录模板序列完全正确。结论:成功构建了针对人蛋白激酶CK 2α亚基的RNA i体系。Objective： To construct a shRNA expressing plasmid targeted against CK2α. Methods.. The templates of shRNAs were chemically synthesized and cloned into the mU6pro vector in the downstream of the U6 promoter. The positive clones were screened by agarose electrophoresis, and the inserted sequence in the recombinants was detected by DNA sequencing. Results： The shRNA expression plasmid was obtained, and the inserted sequence was correct. Conclusion; The RNAi system expressing shRNA targeted against CK2α is successively developed and can be applied to study CK2 function.

关 键 词：蛋白激酶CK2 RNA干扰(RNAI) 短发夹状RNA(shRNA) 基因测序 转染 

分 类 号：Q55[生物学—生物化学] Q34