›› 2012, Vol. 3 ›› Issue (4): 181-184.

• 论著 • 上一篇    下一篇

HDAC8过表达慢病毒载体构建及其对骨髓基质细胞转染及表达的研究

傅瑜1,张平1,张永栋1,单腾飞2,董伟杰1,江宏兵1   

  1. 1. 江苏省南京医科大学附属口腔医院口腔颌面外科
    2. 南京医科大口腔医学研究所;南京医科大学附属口腔医院口腔颌面外科
  • 收稿日期:2012-10-23 修回日期:2012-12-02 出版日期:2012-12-25 发布日期:2013-01-10
  • 通讯作者: 江宏兵 E-mail:jhbtooth@163.com
  • 基金资助:
    表观遗传修饰不同胚源MSCs促进颌骨成骨修饰及相关机制研究

Construction of HDAC8 gene over-expression lentivirus vector and transfection on bone marrow stromal cells

  • Received:2012-10-23 Revised:2012-12-02 Online:2012-12-25 Published:2013-01-10

摘要: 目的 构建大鼠组蛋白去乙酰化酶8(Histone deacetylase 8,HDAC8)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HDAC8的表达。方法 采用DNA重组技术将HDAC8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pGC-FU中,重组获得慢病毒载体pGC-FU-HDAC8。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HDAC8表达情况。结果 测序结果证实HDAC8基因正确插入载体中,成功构建大鼠HDAC8基因过表达载体。Western blot检测显示转染后HDAC8蛋白表达显著上调。结论 针对大鼠HDAC8基因过表达慢病毒载体构建成功,并能有效增强 BMSCs 中HDAC8基因的表达。

Abstract: Objective To construct overexpression vectors targeting rat HDAC8 gene and study the transfection effect of HDAC8 in bone marrow stromal cells ( BMSCs ). Methods The HDAC8 gene was insert into plasmid pGC-FU of lentiviral vector. The plasmid was detected by DNA sequencing. The recombined plasmid pGC-FU-HDAC8 was transfected into 293 cell line. Virus yielded by 293T cell was transfected into rat BMSCs. The expression of HDAC8 was detected by Western bolt. Results The HDAC8 were correctly cloned into the pGC-FU plasmid and confirmed by DNA sequencing. Our result of Western blot showed that pGC-FU-HDAC8 significantly up regulated the expression of HDAC8 in rat BMSCs. Conclusion overexpression vector targeting rat HDAC8 gene had been constructed successfully and efficiently up-regulated the expression of HDAC8 in rat BMSCs.

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