结缔组织生长因子小干扰RNA的构建及其干扰效果检测  

Construction of Small Interfering RNA of Connective Tissue Growth Factor and its Interference Effect

作　　者：李勤操[1] 郑喜邦[2] 王晓晖[1] 杨智洪[1] 姜艳超[1] 丁丽华[1] 李杰萍[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]广西大学动物科学技术学院,广西南宁530005

出　　处：《生物技术通讯》2008年第3期347-349,共3页Letters in Biotechnology

基　　金：国家自然科学基金项目(30370738;30500091;30530320)

摘　　要：目的:构建结缔组织生长因子(CTGF)基因的小干扰RNA(siRNA),并检测其对CTGF基因表达的干扰效果。方法:根据CTGF基因序列,设计2条合理的CTGF-siRNA,并将其克隆到siRNA载体pSliencer2.1-U6 neo中,转化大肠杆菌DH5α,挑取阳性菌落进行酶切和测序鉴定;将构建成功的CTGF-siRNA重组质粒与带FLAG标签的CTGF表达载体共同转染人胚肾293T细胞,Western印迹检测siRNA的干扰效果。结果:构建了2个CTGF-siRNA重组质粒,2条siRNA都有干扰作用,其中一条的干扰效果可达75%以上。结论:构建的CTGF-siRNA可为进一步研究CTGF的功能提供参考。Objective： To construct small interfering RNA（siRNA） of connective tissue growth factor（CTGF） expression vector, and investigate its interference effect. Methods： Two CTGF-siRNA were designed and inserted into the pSliencer 2.1-U6 neo vector. After confirmation of the constructs by restriction and DNA sequencing, human embryo kidney 293T cells were co-transfected with the recombination plasmids and FLAG-CTGF expression vector. The interference effect of CTGF-siRNA was identified by Western blot. Results： Two CTGF-siRNA expression vectors were obtained and could effectively inhibit the expression of CTGF protein, one of which could inhibit the expression over 75%. Conclution： The CTGF-siRNA expression vectors constructed can provide tools for further study on function of CTGF.

关 键 词：结缔组织生长因子 小干扰RNA 表达载体 

分 类 号：Q78[生物学—分子生物学]