14-3-3基因真核表达载体的构建  

Construction of the Eukaryotic Expression Vector of 14-3-3

作　　者：苗新东[1] 刘同美[1] 王婷[1] 陈安琪[1] 崔晓栋[1] 郭军堂[1] 

机构地区：[1]潍坊医学院病理生理学教研室,山东潍坊261053

出　　处：《湖北民族学院学报（医学版）》2009年第4期10-13,共4页Journal of Hubei Minzu University(Medical Edition)

摘　　要：目的克隆人14-3-3基因并构建真核表达载体pcDNA3.1(+)-14-3-3。方法从胎脑组织中提取总RNA,用RT-PCR方法获得人14-3-3基因的全长cDNA,将其插入pCUm-T载体中;序列测定后,重组携带14-3-3基因的表达载体pcDNA3.1(+)-14-3-3。结果经限制性内切酶酶切分析、PCR和DNA序列测定证实目的基因已插入重组质粒。结论成功构建了14-3-3真核表达载体pcDNA3.1(+)-14-3-3,为进一步研究14-3-3在帕金森病中的作用奠定了良好的基础。Objective To clone human 14-3-3 gene and construct the eukaryotic expression vector of pcDNA3.1（＋）-14-3-3.Methods Total RNA was isolated from human brain tissue.The full-length human14-3-3 cDNA was obtained by RT-PCR and then inserted into pCUm-T vector for sequencing.The correct gene was subcloned into pcDNA3.1（＋） to generate recombinant eukaryotic expression vector pcDNA3.1（＋）-14-3-3.Results Enzyme digestion analysis PCR and sequencing showed that the target gene was cloned into recombinant vector.Conclusion The eukaryotic expression vector of pcDNA3.1（＋）-14-3-3 was constructed successfully,which lay the foundation for the further study of 14-3-3 gene in Parkinson disease.

关 键 词：14-3-3基因 真核表达 帕金森病 

分 类 号：R742.4[医药卫生—神经病学与精神病学]