利用RNAi抑制FHL2基因的表达  

Inhibition of the Expression of FHL2 by RNA Interference

作　　者：王朝云[1] 周蕾[1] 丁丽华[1] 吕秋军[2] 杨晓[1] 叶棋浓[1] 

机构地区：[1]军事医学科学院生物工程研究所,北京100850 [2]军事医学科学院放射医学研究所,北京100850

出　　处：《细胞与分子免疫学杂志》2010年第11期1149-1151,共3页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金资助项目(30370738;30530320)

摘　　要：目的:构建FHL2基因的siRNA真核表达载体,观察其对FHL2蛋白表达的影响。方法:利用RNAi干扰技术,设计并合成了两条针对FHL2基因的siRNA,将其克隆到siR-NA表达载体pSliencer 2.1-U6 neo上。将重组质粒和带FLAG标签的FHL2共转染293T人胚肾细胞,通过Western blot实验检验RNAi干扰效应。结果:经过酶切和测序证明,成功构建了FHL2 siRNA真核表达载体。通过Western blot证明,构建的siRNA能有效抑制FHL2基因的表达。结论:成功地构建了FHL2基因的siRNA真核表达载体,转染细胞后能有效地抑制FHL2基因的表达。AIM： To construct the eukaryotic expression vector of small interfering RNA（siRNA） targeting human FHL2 and transfect it into human embryo kidney 293T cells to investigate the silencing effect of FHL2 siRNA on the expression of FHL2 gene.METHODS： Two FHL2 siRNAs were designed and inserted into pSliencer 2.1-U6 neo expression vector.Then human embryo kidney 293T cells were cotransfected with the recombinant plasmids and FLAG-tagged FHL2 expression vector.The silencing effect of FHL2 siRNAs on the expression of FHL2 gene was identified by Western blot.RESULTS： The expression vectors of FHL2 siRNAs were constructed and confirmed by DNA sequencing.Western blot showed that FHL2 siRNAs effectively inhibited expression of FHL2.CONCLUSION： The eukaryotic expression vectors of FHL2 siRNAs are constructed successfully.The siRNAs effectively inhibit the expression of FHL2.

关 键 词：FHL2 SIRNA 基因表达 

分 类 号：R392.11[医药卫生—免疫学]