摘要
目的:设计并构建TIN-ag-RP基因的小干扰RNA(siRNA),检测其对小管间质性肾炎抗原相关蛋白(TIN-ag-RP)表达的干扰效果。方法:设计2条针对TIN-ag-RP基因的siRNA,并克隆到siRNA表达载体pSliencer 2.1-U6 neo上;经酶切和测序证明构建成功后,将重组质粒和带FLAG标签的TIN-ag-RP基因共转染293T人胚肾细胞,通过Western blot检验siRNA的干扰效果。结果:获得了2个TIN-ag-RP基因siRNA真核表达载体,均能有效抑制TIN-ag-RP的表达,其中一条的抑制效率达90%以上。结论:构建的TIN-ag-RP基因的siRNA能有效抑制TIN-ag-RP的表达。
Objective： To design and construct the small interference RNA（siRNA） for TIN-ag-RP gene and investigate the interference effect on the expression of tubulointerstitial nephritis antigen related protein（TIN-ag-RP）. Methods： Two TIN-ag-RP siRNAs were designed and inserted into pSliencer 2.1-U6 neo expression vector. After confirmation of the constructs by restriction digestion and DNA sequencing, human embryo kidney 293T cells were cotransfected with the re- combination plasmids and FLAG-tagged TIN-ag-RP expression vector. The effect of TIN-ag-RP siRNAs on the expression of TIN-ag-RP was identified by Western blot. Results： The two TIN-ag-RP siRNAs were obtained and could effectively inhibit the expression of TIN-ag-RP, one of which could inhibit the expression up to 90% or more. Conclusion： The constructed TIN-ag-RP siRNAs could effectively inhibit the expression of TIN-ag-RP.
出处
《生物技术通讯》
CAS
2008年第2期185-187,共3页
Letters in Biotechnology
基金
国家自然科学基金项目(30530320
30625035)
全军"十一五"医药卫生科研基金项目(06J021)
关键词
小管间质性肾炎抗原相关蛋白
小干扰RNA
基因表达
tubulointerstitial nephritis antigen related protein
small interference RNA
gene expression
