FTH1真核表达载体的构建与鉴定  

Construction and Identification of FTHl Gene Eukaryotic Expression Vector

作　　者：董晓[1] 何淑雅[1] 李斌元[1] 王三虎[1] 自映萍[1] 马云[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：《中南医学科学杂志》2011年第5期487-490,共4页Medical Science Journal of Central South China

基　　金：国家自然科学基金项目(30370795);湖南省自然科学基金(07JJ3030)资助项目

摘　　要：目的构建携带人铁蛋白重链多肽1(FTH1)基因的真核表达载体pCMV-Myc-FTH1,为研究FXR1P与FTH1体内相互作用奠定基础。方法以人胎脑cDNA文库为模板进行聚合酶链反应(PCR)特异扩增FTH1基因片段,将扩增片段经EcoRⅠ和XhoⅠ双酶切后克隆到同样经EcoRⅠ和XhoⅠ双酶切的pCMV-Myc载体,酶切鉴定阳性克隆并进一步测序鉴定。结果成功构建了pCMV-Myc-FTH1重组真核表达载体。结论 pCMV-Myc-FT1重组真核表达载体的构建为进一步在细胞内鉴定FXR1P与FTH1相互作用奠定了基础,对研究FXR1在脆性X综合征中的作用机理提供了依据。Objective To construct a eukaryotic expression vector pCMV-Myc-FTH1 that can express the human ferritin heavy peptide 1 （FTH1） in order to study the interaction between FXR1P and FTH1. Methods The FTH1 gene was amplified from a human fetal brain pGADT7-cDNA library plasmid, digested by EcoR I and Xho I and then cloned into eukaryotic expression vector pCMV-Myc which was also digested by EcoR I and Xho I. The recombinant plasmid of pCMV- Myc-FTHI was identified by enzyme digestion and sequence analysis. Result The recombinant eukaryotic expression vector pCMV-Myc-FTH1 was successfully constructed. Conclusion The construction of recombinant eukaryotic expression vector pCMV-Myc-FTH1 will lay the foundation further for the identification of the interaction between FXR1P and FTH1 ,and provide some basis for the study of the mechanism of FXR1 within fragile X syndrome.

关 键 词：脆性X综合征 FTH1 真核表达载体 

分 类 号：Q78[生物学—分子生物学]