›› 2013, Vol. 4 ›› Issue (2): 61-64.

• 论著 • 上一篇    下一篇

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

宋辉,江宏兵,董伟杰,张平,戈杰   

  1. 江苏省南京医科大学附属口腔医院口腔颌面外科
  • 收稿日期:2013-03-27 修回日期:2013-06-06 出版日期:2013-06-25 发布日期:2013-06-26
  • 通讯作者: 江宏兵 E-mail:jhbtooth@163.com
  • 基金资助:
    国家自然科学基金

Construction of STAB2 gene over-expression lentivirus vector and transfection on bone marrow stromal cells

  • Received:2013-03-27 Revised:2013-06-06 Online:2013-06-25 Published:2013-06-26

摘要: 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。

Abstract: 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 293 T 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.

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