基于smad7基因为靶序列的RNA干涉作用研究  

THE STUDY ON RNA INTERFERENCE BASED ON THE TARGET SEQUENCE OF SMAD7 GENE

作　　者：王菊萍[1] 王莹[2] 霍艳英[3] 郭蔼光[1] 张开泰[3] 

机构地区：[1]西北农林科技大学生命科学学院,杨凌712100 [2]重庆医科大学基础医学院,重庆400016 [3]军事医学科学院放射医学研究所分子毒理研究室,北京100850

出　　处：《内蒙古农业大学学报（自然科学版）》2004年第4期21-25,共5页Journal of Inner Mongolia Agricultural University(Natural Science Edition)

基　　金：国家重点基础研究发展规划973项目(G1998051207)

摘　　要：　本文旨在探讨小干涉RNA(siRNA)对靶基因Smad7mRNA表达的阻断作用。用体外转录方法制备Smad7基因的小干涉RNAs(siRNAs),脂质体介导瞬时转染BERP35T-2肺癌细胞系,Northemblot杂交检测靶基因mRNA的表达丰度。根据Smad7编码区序列在体外成功地制备了针对两个不同靶序列的siRNAs,Northemblot杂交显示,在转染siRNA和BERP35T-2细胞中,不管是内源性的还是外源性的Smad7mRNA的表达丰度均明显下降,Smad7基因编码区中542bp～563bp及701bp～722bp两个区域均是siRNA作用的有效靶序列;本研究设计并制备的siRNAs能有效抑制smad7基因的表达。The suppression of siRNA to the expression of the mRNA of target gene Smad7 was studied.The siRNAs of Smad7 gene were prepared by in virto transcription.BERP35T-2 lung cancer cell line was transient transfected with siRNAs and Smad7 expression plasmid using Lipofectamine 2000 reagent.The expression abundance of target gene mRNA was tested with Northern blot hybridization.According to the coding sequence of Smad7 gene, the siRNAs for two different target sequences in vitro were syntnesized successfully. The result of Northem blot hybridization showed that the expression abundance of both endogenous and exogenous Smad7 mRNA decreased evidently in BERP35T-2 cells transfected with siRNAs. Both regions of 542bp～563bp and 701bp～722bp in the coding region of Smad7 gene are effective target sequences which can be acted by siRNA.The siRNAs designed and synthesized in our research can suppress the expression of Smad7 gene effectively.

关 键 词：SMAD7基因 小干涉RNA SMAD7 表达 作用研究 靶基因 转染 RNA干涉 序列 编码区 

分 类 号：S852.651[农业科学—基础兽医学] R575.2[农业科学—兽医学]