CAJ | 학술논문

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。
목적：구건병감정sox4사단불동결구역적중조진핵표체질립。방법：이HL-60세포적총RNA위모판，용RT-PCR방법확증출sox4기인cDNA，장산물극륭진진핵재체pCMV-Flag질립내，구건함sox4불동결구역적중조진핵표체질립。장pCMV-Flag-sox4중조질립전염293T세포，Western blot검측sox4단백표체。결과：핵산서렬분석sox4이성공삽입pCMV-Flag재체중，전염pCMV-Flag-sox4적293T세포중검측도표체적sox4단백。결론：성공구건료함sox4기인적중조진핵표체질립。
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.