摘要
目的:构建靶向人分化相关基因NDRG1的siRNA表达载体。方法:设计靶向人NDRG1发夹样siRNA,依据设计,合成2条互补的寡核苷酸链,退火后连接入pSUPERneo-GFP,转化扩增后得到重组载体pSUPERneo-GFP-siNDRG1,进行序列测定。用脂质体转染法将重组载体转染食管癌细胞株EC9706,采用RT-PCR检测转染细胞NDRG1mRNA的表达水平。结果:经过酶切鉴定与测序,pSUPERneo-GFP-siNDRG1构建成功。稳定转染该重组体的EC9706细胞NDRG1mRNA表达水平降低。结论:成功构建了针对NDRG1基因的siRNA表达载体。
Aim：To construct NDRG1 gene-targeted small interfering RNA（siRNA） vector.Methods：NDRG1 gene-targeted hairp in siRNA was designed,then two complementary oligo nucleotide strand were synthesized and inserted into pSUPERneo-GFP vector after annealing to get the new vector pSUPERneo-GFP-siNDRG1,which was digested by restrictive enzyme and sequenced.Then the new vector was transfected into human esophageal carcinoma EC9706 cell using lipofectamine method,and the NDRG1 mRNA expression level was detected by RT-PCR.Results and Conclusion ： pSUPERneo-GFP-siN-DRG1 was built successfully, and the expression level of NDRG1 mRNA in transfected EC9706 cells decreased.
出处
《郑州大学学报（医学版）》
CAS
北大核心
2009年第4期700-702,共3页
Journal of Zhengzhou University(Medical Sciences)
基金
国家自然科学基金资助项目30470779
