Satb2过表达慢病毒载体构建及其对骨髓基质细胞的转染及表达

Construction of Satb2 gene over-expression lentivirus vector and transfection on bone marrow stromal cells

目的:构建大鼠特殊富含AT序列结合蛋白2(special AT-rich binding protein 2,Satb2)基因过表达的慢病毒载体,转染大鼠骨髓基质细胞(bone marrow stromal cells,BMSCs)后观察Satb2的表达。方法:采用DNA重组技术将Satb2基因插入到含有绿色荧光蛋白(GFP)基因的慢病毒表达载体质粒GV208中,获得重组慢病毒载体GV208-Satb2。重组慢病毒载体经过测序鉴定后转染293T细胞生产病毒液,用得到的病毒液转染大鼠BMSCs,Western blot分析转染前后Satb2表达情况。结果:测序结果证实Satb2基因正确插入载体中,成功构建大鼠Satb2基因过表达载体。Western blot检测显示转染后Satb2蛋白表达显著上调。结论:针对大鼠Satb2基因过表达慢病毒载体构建成功,并能有效增强BMSCs中Satb2基因的表达。

Objective：To construct overexpression vectors targeting rat Satb2 gene and study the transfection effect of Satb2 in bone marrow stromal cells（ BMSCs） .Methods：The Satb2 gene was insert into plasmid GV208 .The plasmid was detected by DNA sequencing.The recombined plasmid GV208-Satb2 was transfected into 293T cell line.Virus yielded by 293T cell was transfected into rat BMSCs.The expression of Satb2 was detected by Western bolt.Results：The Satb2 was correctly cloned into the GV208 plasmid and confirmed by DNA sequencing.The result of Western blot showed that GV208-Satb2 significantly up-regulated the expression of Satb2 in rat BMSCs.Conclusions：Over-expression vector targeting rat Satb2 gene had been constructed successfully and efficiently up-regulated the expression of Satb2 in rat BMSCs.