靶向人Pokemon基因的shRNA重组质粒的构建  

Construction of the hairpin RNA recombinant plasmids targeting human Pokemon gene

作　　者：曹文淼[1] 任精华[1] 姚杰[1] 张涛[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院肿瘤中心,湖北武汉430023

出　　处：《世界肿瘤杂志》2009年第1期33-36,共4页Tumour Journal of the World

摘　　要：目的构建靶向Pokemon基因的短发夹RNA（shRNA）真核表达载体，为研究Pokemon基因在肿瘤发生发展中的作用奠定基础。方法利用基因重组技术，依据siRNA的设计原则，针对目的基因的mRNA序列筛选了3对siRNA序列以及一对阴性对照序列，将其分别插入真核表达载体psiRNA-hHlneo H1启动子下游，经α互补筛选、酶切和DNA测序鉴定，构建靶向Pokemon基因的RNAi真核表达载体。结果酶切鉴定及DNA测序结果显示siRNA片断正确，与设计序列一致，且成功插入预定位点。结论靶向Pokemon基因的shRNA重组质粒成功构建，为进一步研究Pokemon基因在多种疾病中的作用以及探索肿瘤的基因治疗奠定基础。Objective To establish the foundation for studying the role of Pokemon gene in tumorigenesis and development by constructing recombinant plasmids that can express small interfering RNA （siRNA） targeting human Pokemon gene. Methods Hairpin siRNA templates targeting Pokemon gene were synthesized and cloned into plasmid vector psiRNA-Hlneo. Three vectors derived siRNAs （psiRNA1, 2 and 3） and one mocking psiRNAc （as control） were constructed. The recombinant Pokemon siRNA plasmids were constructed and identified using restrictive enzyme analysis and DNA sequencing. Results Restrictive enzyme analysis and DNA sequencing revealed the successful construction of siRNA expression plasmids. Conclusions Constructing siRNA templates targeting Pokemon gene may provide us with practical tools for further study the role of Pokemon gene in the development of some diseases and gene therapy of tumor.

关 键 词：POKEMON RNAI SHRNA 

分 类 号：R730.54[医药卫生—肿瘤]