PS1基因真核表达载体的构建与鉴定  

Construction and identification of eukaryotic expression vector of PS1 gene

作　　者：宁丽峰[1] 王慧萍[1] 李铁[2] 桑建利[2] 李佳慧[3] 

机构地区：[1]国家人口计生委科学技术研究所,北京100081 [2]北京师范大学生命科学院细胞生物学研究所,北京100875 [3]中国医学科学院中国协和医科大学实验动物研究所,北京100021

出　　处：《哈尔滨医科大学学报》2011年第2期117-119,123,共4页Journal of Harbin Medical University

基　　金：科学技术部"科技基础工作项目"(G99-E-01)

摘　　要：目的构建PS1基因真核表达载体,为PS1基因功能研究提供工具。方法从C57BL/6小鼠不同胚胎中提取总RNA,采用反转录-聚合酶链反应扩增PS1基因编码区,将其克隆入pMD18-T载体中。通过聚合酶链反应,酶切和DNA测序鉴定。结果 PS1在9日龄小鼠胚胎的cDNA中高表达。测序显示融合基因GFP-PS1和PS1-GFP中PS1为全长PS1编码序列。Western blot检测显示融合蛋白GFP-PS1与PS1-GFP的相对分子质量均为77 kD左右。结论成功地构建了PS1基因真核表达载体。Objective To construct an eukaryotic expression vector for presenilin 1（PS1） gene and to provide a tool for studying of PS1 gene function.Methods The total RNAs were isolated from various embryos of C57BL/6 mice.The cDNA of PS1 gene was amplified by reverse transcription-polymerase chain reaction.After purification,the gene was cloned into pMD18-T vector.The recombinant plasmid was identified by enzyme digestion,DNA sequencing and Western blot.Results The PS1 was highly expressed in 9-day embryo.Sequencing showed that the PS1 coding sequence of GFP-PS1 and PS1-GFP was full-length.Western blot showed that the molecular weight of the fusion protein of GFP-PS1 and PS1-GFP was about 77×103 kD.Conclusion The eukaryotic expression vector of PS1 gene has been successfully constructed,which may provide a basis for further researches.

关 键 词：早老素1基因 小鼠 胚胎 基因克隆 真核表达 

分 类 号：R349.64[医药卫生—基础医学]