信号转录激活子3基因shRNA慢病毒载体的构建与鉴定  

Construction of STAT3 shRNA lentiviral vector

作　　者：阮坚丽[1] 高源[1] 管文贤[1] 乔志明[1] 钱伟峰[1] 

机构地区：[1]南京医科大学附属苏州市立医院急诊科,苏州215002

出　　处：《南通大学学报（医学版）》2010年第2期83-85,共3页Journal of Nantong University(Medical sciences)

基　　金：苏州市社会发展计划项目(SS08033)

摘　　要：目的:构建人信号转录激活子3(STAT3)基因shRNA慢病毒载体。方法:从GenBank STAT3 mRNA上寻找到3条有效靶序列,合成3对针对靶序列的siRNA oligo,通过筛选获得最佳靶序列后合成相应的shRNA模板,将合成好的两条互补的DNA单链退火形成双链DNA,经与BamH Ⅰ和Xho I酶切的pRNAT-U6.2/Lenti载体连接后产生shRNA慢病毒载体,酶切和测序鉴定。结果:酶切和测序鉴证实合成STAT3shRNA慢病毒载体寡核甘酸链插入正确。结论:成功构建人STAT3基因shRNA慢病毒载体。Objective：To construct a reco mbinant lentiviral vector of RNA interference of STAT3 gene.Methods：Three different siRNAs of STAT3 were designed and synthesized.After being screened,the most effective siRNA was found.According to this sequence,the short hairpin DNA of STAT3 was constructed.The shRNA duplex was ligated into the recombinant vector pRNAT-U6.2/Lenti.The recombine vector was confirmed by enzyme cutting and DNA sequencing.Results：The restriction map and DNA sequencing demonstrated that the recombinant lentiviral vector of RNA interference of STAT3 gene was constructed correctly.Conclusion：The lentivirus RNAi vector targeting STAT3 has been successfully constructed,which will provide a tool for the further study on function of STAT3 gene in tumors.

关 键 词：信号转录激活子3 RNA干扰 慢病毒 

分 类 号：R394[医药卫生—医学遗传学]