中文摘要：

目的：构建大鼠组蛋白去乙酰基转移酶（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，实时荧光定量PCR和蛋白质免疫印迹法分析转染前后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。大鼠BMSCs转染后HDAC8 mRNA及蛋白表达显著上调。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。

英文摘要：

Objective：To construct rat HDAC8-gene-over-expressed lentivirus vector and study expression of HDAC8 in bone marrow stromal cells （ BMSCs ） after transfection. Methods：The HDAC8 gene was inserted into plasmid pGC-FU of lentiviral vector by recombinant DNA technology. Lentivirus vector pGC-FU-HDAC8 was got after recombination. The The recombinant lentivirus vector 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 and real-time RT-PCR before and after transfection. Results：It was confirmed by DNA sequencing that the HDAC8 was correctly inserted into the vector, and that rat HDAC8-gene-over-expressed lentivirus vectors were successfully constructed. After transfection, the expression of HDAC8 was significantly upregulated either in mRNA level or in protein lever in rat BMSCs. Conclusions：The construction of HDACS-gene-over-expressed lentivirus vector was successful, and it efficiently up-regulated the expression of HDAC8 in rat BMSCs.