sox4不同结构域的真核表达质粒的构建和鉴定  

Construction and identification of eukaryotic expression plasmids of sox4 different domains

作　　者：史露露[1] 宋冠华[1] 潘素飞[1] 禹林昌 史美燕 任霞[1] 郭强[1] 姜国胜[1] 

机构地区：[1]山东省医学科学院基础医学研究所济南大学,山东济南250062

出　　处：《医学检验与临床》2014年第1期3-5,共3页Medical Laboratory Science and Clinics

基　　金：基金资助：国家自然科学基金（项目编号：30810444）;国家自然科学基金（项目编号：30771103）;国家自然科学基金（项目编号：81172792）;省自主成果转化重大专项（项目编号：2012ZHZX1A0420）;省科技攻关重点项目（项目编号：2006GG2302010）;省科技攻关重大项目（项目编号：2007GG2002023）

摘　　要：目的：构建并鉴定sox4四段不同结构域的重组真核表达质粒。方法：以HL-60细胞的总RNA为模板，用RT—PCR方法扩增出sox4基因cDNA，将产物克隆进真核载体pCMV—Flag质粒内，构建含sox4不同结构域的重组真核表达质粒。将pCMV—Flag—sox4重组质粒转染293T细胞，Westernblot检测sox4蛋白表达。结果：核酸序列分析sox4已成功插入pCMV—Flag载体中，转染pCMV—Flag-sox4的293T细胞中检测到表达的sox4蛋白。结论：成功构建了含sox4基因的重组真核表达质粒。Objective ： To construct and identify recombinant eukaryotic expression plasmids containing sox4 gene. Methods ： The cDNA sox4 gene was amplified by RT-PCR with the total RNA in HL-60 cells as a template .The PCR fragments were cloned into p-Flag vector to construct recombinant eukaryotic expression plasmids containing sox4 gene.The p-Flag-sox4 recombinant plasmid were respectively transfected into 293T cells and the expression plasmid of sox4 protein was detected by Western blot.Results ： The DNA sequence and double restrictive endonuclease analysis showed that sox4 was successfully inserted into pCMV-Flag vector. The expression of sox4 protein was detected in293T cells transfected with pCMV- Flag-sox4 plasmid by Western blot. Conclusions ： Recombinant eukaryotic expression plasmids containing sox4 gene has been successfully constructed.

关 键 词：pCMV—Flag sox4 HL-60 

分 类 号：R378.911[医药卫生—病原生物学]