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 sequen-cing.The recombined plasmid GV208-Satb2 was transfected into 293T cell line.Virus yielded by 293T cell was transfected into rat BM-SCs.The expression of Satb2 was detected by Western bolt.Results:The Satb2 was correctly cloned into the GV208 plasmid and con-firmed 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.
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