中国医科大学学报2012，Vol.41Issue(7)：580-584,5.
双表达骨形态发生蛋白9、7重组腺病毒载体的构建及其对间充质干细胞C3H10的成骨作用
Construction of Recombinant Adenovirus Co-expressing BMP9, BMP7 and Effect on Osteogenesis of Mesenchymal Stem Cells C3H10
摘要
Abstract
Objective To construct a recombinant adenovirus co-expressing BMP9 and BMP7, and identify its primary effect on osteogenesis. Methods The gene sequence of BMP9 and BMP7 were amplified from AdEasy vector by PCR and sub-cloned into pASG2 vector. The co-expression shuttle plasmid pASG2-BMP9,7 was confirmed by restriction endonuclease digestion,PCR and gene sequencing;then plasmid pAdeasy-1 was co-transfected to produce recombinant adenovirus plasmid by homologous recombination. In addition,the recombinant vector was transfected into HEK293 cells. High-titer recombinant adenovirus (Ad-BMP9, 7) was collected after rounds of amplification. The capability of osteogenesis induced with Ad-BMP9,7 was evaluated in vitro in C3H10 cells by alkaline phosphatase staining and calcium-alizarin red staining. Results Ad-BMP9, 7 was constructed successfully. Testing with C3H10 cells,alkaline phosphatase staining and calcium-alizarin red staining were stronger than single transfected BMP7 or BMP9 cells. Conclusion The recombinant adenovirus co-expressing BMP9 and BMP7 was constructed successfully .which has a capability of inducing osteogenic differentiation of C3H10 cells.关键词
双表达BMP9、7载体/重组腺病毒/载体构建/成骨Key words
co-expressing of BMP9 and BMP7/recombinant adenovirus/construction/osteogenesis分类
医药卫生引用本文复制引用
罗庆,陈聪,迭小红,田雯,康权..双表达骨形态发生蛋白9、7重组腺病毒载体的构建及其对间充质干细胞C3H10的成骨作用[J].中国医科大学学报,2012,41(7):580-584,5.基金项目
国家自然科学基金资助项目(30973062) （30973062）
重庆市自然科学基金资助项目(CSTC2009BB5408) （CSTC2009BB5408）
