sox4不同结构域的真核表达质粒的构建和鉴定

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，Western blot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.