Smad4-siRNA真核表达载体的构建与鉴定  

Construction and Identification of Eukaryotic Expression Vector of RNA Interference of Smad4 Gene

作　　者：李萍[1] 范玉强[1] 曹武奎[1] 梁树人[1] 李顺天[1] 郄春花[1] 戴晨阳[1] 李秀梅[1] 徐健[1] 刘莉[1] 

机构地区：[1]天津市传染病医院天津市肝病研究所,天津300192

出　　处：《实用预防医学》2007年第4期1028-1030,共3页Practical Preventive Medicine

摘　　要：目的构建Smad4特异的RNA干扰质粒载体,为探讨抑制Smad4表达在肝纤维化治疗中的意义奠定基础。方法根据GenBank数据库提供的Smad4基因核苷酸序列,选择设计能转录小发夹结构RNA(Small hairpin RNAs,shRNA)的DNA序列,并与psilencer3.1-H1-hygro质粒载体连接,构建真核表达载体,使用限制性内切酶鉴定及测序鉴定是否为阳性克隆。结果成功构建psilencer3.1-Smad4载体,拟进一步采用RNAi技术观察其对Smad4基因表达的抑制情况,从而研究其在肝纤维化治疗中的作用。结论Smad4-siRNA真核表达载体被成功构建。Objective To construct eukaryotic expression vector of RNA interference specific for Smad4, and to explore the significance of Smad4 inhibition in treatment of hepatic fibrosis. Methods Genome sequences of Smad4 gene was retrieved from Genbank and cDNA was designed coding expression of shRNA （small hairpin RNAs） for Smad4 gene. The cDNA was synthesized and inserted into psilencer3.1 - H1 - hygro which was a eukaryotic expression vector and identified by the restriction map and sequencing. Results The recombinant plasmid of RNA interference specific for Smad4 was identificated by the restriction map and sequencing which was completely coincided with the designs. Conclusions The siRNA eukaryotic expression vector against Smad4 mRNA has been successfully constructed.

关 键 词：SMAD4 RNA干扰 肝纤维化 载体构建 

分 类 号：Q522[生物学—生物化学]