靶向Annexin A3短发夹环RNA干扰载体的构建及鉴定  

Construction and Identification of shRNA Eukaryotic Expression Vector Targeting Annexin A3

作　　者：席仁荣[1] 刘建军[1] 吴振强[2] 邢秀梅[1] 黄海燕[1] 

机构地区：[1]深圳市疾病预防控制中心毒理研究室,广东深圳518020 [2]华南理工大学生物科学与工程学院,广东广州518640

出　　处：《安徽农业科学》2008年第17期7145-7146,7246,共3页Journal of Anhui Agricultural Sciences

基　　金：国家自然科学基金资助项目(30571557);广东省自然科学基金资助项目(5009153)

摘　　要：[目的]利用RNA干扰(RNA interfering,RNAi)技术构建和鉴定膜联蛋白A3(Annexin A3)的真核表达载体,为进一步研究Annexin A3在肝细胞中的功能奠定基础。[方法]人工合成3对shRNA序列,退火后定向克隆到真核表达载体psiRNA-hH1neo上,采用酶切和测序法鉴定。[结果]质粒酶切及测序鉴定得到的产物与预期的目的基因一致。[结论]成功构建膜联蛋白A3靶向的真核表达载体。[Objective] The aim of this paper was to construct and identify the siRNA eukaryotic expression vector targeting annexin A3. [Methods] Four shRNAs were designed according to the coding sequence of annexin A3 gene, and cloned into the downstream of H1 promoter of psiRNA-hH1 -neo, The constructed recombinant was analyzed and identified by Ase I endonuclease digestion and DNA sequencing. [Results] The constructed psiRNA plasmid digested with Ase I was linearized. The sequencing result confirmed that the sequence of inserted fragment was correct. [Conclusion] Eukaryotic expression vector of siRNA targeting annexin A3 gene was successfully constructed.

关 键 词：ANNEXIN A3 psiRNA—hHlneo RNA干扰 真核表达载体 

分 类 号：Q782[生物学—分子生物学]