FXR1真核表达载体的构建及鉴定  

Construction and Identification of Eukaryotic Expression Vector of FXR1

作　　者：杨阳[1] 马云[1] 符向辉[1] 何淑雅[1] 

机构地区：[1]南华大学生物化学与分子生物学教研室,湖南衡阳421001

出　　处：《南华大学学报（医学版）》2010年第1期27-30,共4页Journal of Nanhua University(Medical Edition)

基　　金：湖南省自然科学基金(07JJ3030);湖南省卫生厅(B2006-099)资助项目

摘　　要：目的构建携带人脆性X相关基因1(FXR1)的真核表达载体pCMV-HA,为研究FXR1P互作蛋白奠定基础。方法以pYESTrp3-FXR1为模板进行聚合酶链反应(PCR)特异扩增FXR1基因片断,将扩增片段经EcoR I和XhoI双酶切后克隆到pCMV-HA载体,酶切鉴定阳性克隆并测序鉴定。结果成功构建了pC-MV-HA/FXR1真核表达载体。结论pCMV-HA/FXR1真核表达载体的构建为进一步在细胞内鉴定FXR1互作蛋白奠定了基础,从而对研究FXR1在脆性X综合征中的作用机理具有深远意义。Objective To clone human fragile-X-related gene 1（FXR1）and construct its eukaryotic expression vector in order to study the interactive protein with FXR1P. Methods The FXR1 gene was amplified from pYESTrp3/FXR1 plasmid,digested with EcoRI and XhoI and then cloned into eukaryotic expression vector pCMV-HA.The recombinant plasmid of pCMV-HA/FXR1 was verified by enzyme digestion and sequence analysis. Results The construction of eukaryotic expression vector pCMV-HA/FXR1 was successful and the resulted sequence completely coincided with the designs.Conclusion The construction ofeukaryotic expression vector pCMV-HA/FXR1 will lay a foudation for identifying interactive protein with FXR1P and be helpful for further studies of the mechanisms of FXR1 gene in Fragile X syndrome.

关 键 词：脆性X综合征 FXR1基因 免疫共沉淀 

分 类 号：Q782[生物学—分子生物学]