CRMP-1真核表达载体的构建  

Construction of CRMP-1 Eukaryotic Expression Vector

作　　者：乔宠[1] 常瑞晶[1] 王春辉[2] 

机构地区：[1]中国医科大学附属盛京医院妇产科,沈阳110004 [2]中国人民解放军沈阳军区总医院肝胆外科,沈阳110016

出　　处：《中国医科大学学报》2011年第4期293-296,共4页Journal of China Medical University

基　　金：辽宁省自然科学基金资助项目(20082096)

摘　　要：目的构建人坍塌反应调节蛋白1(CRMP-1)基因表达载体,以研究CRMP-1蛋白在目的细胞中的生物学功能。方法从购买的人胚肾细胞株293T中提取总RNA,经过RT-PCR方法扩增含人CRMP-1基因的全长cDNA,将CRMP-1基因开放阅读框(ORF)cDNA序列,克隆到真核表达载体p3x-FLAG-myc-CMV-24中,构建真核表达质粒CRMP-1-p3x-FLAG-myc-CMV-24。以酶切和基因序列测定方法鉴定重组质粒CRMP-1-p3x-FLAG-myc-CMV-24的正确性。结果酶切鉴定和基因序列测定均证实重组质粒CRMP-1-p3x-FLAG-myc-CMV-24构建正确。结论成功构建人CRMP-1基因真核表达载体。Objective To study the biological function of collapsin response mediator protein-1（CRMP-1）in the cells,we construct the eukaryotic expression vector of human CRMP-1 gene.Methods The total RNA was extracted from the human embryo kidney 293 T cell line.CRMP-1 cDNA was obtained by RT-PCR amplication and the cDNA ORF（open reading frame）was cloned into p3x-FLAG-myc-CMV-24 vector to construct recombinant CRMP-1-p3x-FLAG-myc-CMV-24 plasmid.The correctness of the recombinant CRMP-1-p3x-FLAG-myc-CMV-24 plasmid was tested by the restriction enzyme analysis and gene sequencing method.Results The results of restriction enzyme analysis and gene sequencing method indicated the correctness of the recombinant CRMP-1-p3x-FLAG-myc-CMV-24 plasmid construction.Conclusion Human CRMP-1 eukaryotic expression vector is successfully constructed.

关 键 词：坍塌反应调节蛋白1 基因克隆 真核表达载体 

分 类 号：Q784[生物学—分子生物学]