检验医学与临床Issue(21)：2958-2959,2962,3.DOI:10.3969/j.issn.1672-9455.2014.21.006
新靶点CyclinE干扰RNA真核表达载体的构建
Construction of new target eukaryotic expression vectors of CyclinE siRNA
摘要
Abstract
Objective To construct the 4 new targets eukaryotic expression vectors of human CyclinE interfer-ence specific RNA to transfect into glioma U 251 cells ,the mRNA expression was detected by RT-PCR ,for obtaining the ukaryotic expression vectors with the best interference effect to provide the valuable data for Cyclin E becoming the human tumor marker .Methods (1)Four new targets eukaryotic expression vectors of RNA interference specific CyclinE were constructed ,named as PCyclinE-1 ,PCyclinE-2 ,PCyclinE-3 and PCyclinE-4 .The double digestion method and the base sequence measuring were adopted to detect whether the vectors were successfully constructed .(2)The four newly constructed vectors were transfected into U 251 cell line by Lipofectamine 2000 .(3)The RT-PCR results was adopted to detect the expression quantity of CyclinE mRNA for selecting a vector with best interference .Results (1)4 new targets eukaryotic expression vectors of RNA interference specific for CyclinE were constructed .(2)CyclinEmRNA was obviously suppressed ,the eukaryotic expression vector with best interference effect for human CyclinE interfer-ence specific RNA was obtained .Conclusion The new eukaryotic expression vector of CyclinE siRNA with best effect is successfully constructed ,which significantly suppress the CyclinE mRNA expression of U 251 cells .关键词
干扰RNA/新靶点/细胞周期蛋白E/胶质瘤Key words
interference RNA/new target/CyclinE/glioma引用本文复制引用
渠志臻,刘珊,王艳艳,高乃康,呼格吉乐..新靶点CyclinE干扰RNA真核表达载体的构建[J].检验医学与临床,2014,(21):2958-2959,2962,3.基金项目
内蒙古自然科学基金资助项目（2012MSll47）。 （）
