医学分子生物学杂志2011，Vol.8Issue(6)：471-474,4.DOI:10.3870/j.issn.1672-8009.2011.06.001
慢病毒重组同源盒基因HOXA4表达载体的构建及其感染人脐带间充质干细胞的实验性研究
Construction of Recombinant Lentivirus Containing Human HoxA4 Gene and Their Transfection Efficiency in Human Umbilicalcord Mesenchymal Stem Cells
摘要
Abstract
Objective To construct recombinant lentivirus vector containing human H0XA4 gene and test its transfection in human umbilicalcord mesenchymal stem cells ( hUCMSCs ) . Methods H0XA4 gene was amplified from plasmid H0XA4 - MSCV by PCR technique and sub-cloned into the expression plasmid of lentiviral-GFP-CTB vector. The lentiviral-GFP-H0XA4 expression vector was constructed and confirmed by endoenzyme digestion and sequencing. Recombinant Lentiviral-GFP-HOXA4 was produced by 293 T cells following the co-transfection of lentiviral-GFP-H0XA4 and packaging plasmids pRsv-REV pMDlg-pRRE and PMD2G. The recombinant lentiviral-GFP-H0XA4 was then used to infect hUCMSCs. Results The recombinant lentiviral-GFP-H0XA4 was produced successfully. High-purified virus supernatant was obtained, with virus titer of 2. 11 x 108 TU/ml and efficient infection in hUCMSCs expressing green fluorescence ( 95. 37% ) 60 h after transfection. Conclusion The lentiviruse vector with human H0XA4 cDNA was established successfully, with effective transfection in hUCMSCs.关键词
同源基因HOXA4/慢病毒表达载体/人脐带间充质干细胞Key words
H0XA4 gene/ lentiviral vector/ human umbilicalcord mesenchymal stem cells分类
医药卫生引用本文复制引用
贺玲,贺文凤,郭玲玲,李剑..慢病毒重组同源盒基因HOXA4表达载体的构建及其感染人脐带间充质干细胞的实验性研究[J].医学分子生物学杂志,2011,8(6):471-474,4.基金项目
国家自然科学基金(No.C03030309) （No.C03030309）
