重组骨形态发生蛋白7基因转染骨髓间充质干细胞及其在该细胞中的表达

Expression in bone marrow mesenchymal cells transfected by bone morphogenetic protein 7 gene recombination and its significance

目的将重组骨形态发生蛋白7质粒（pcDNA3．1-BMP7）转染至体外培养的兔骨髓间充质干细胞中．测定骨形态发生蛋白7基因在该细胞中的表达。方法骨形态发生蛋白7基因序列测定、拼接后与报道的骨形态发生蛋白7基因序列对照：脂质体介导pcDNA3．1-BMP7转染骨髓间充质干细胞，G418筛选14d，Western blot、PCR、原位杂交检测骨形态发生蛋白7基因的表达。结果序列测定证实与GenBank报道的骨形态发生蛋白7全长基因序列一致：转染pcDNA3．1-BMP7质粒的骨髓间充质干细胞有大量骨形态发生蛋白7蛋白及骨态发生蛋白7 mRNA表达。结论脂质体介导下，重组真核表达载体pcDNA3．1-BMP7能被转入骨髓间充质干细胞．且表达了骨形态发生蛋白7及骨形态发生蛋白7 mRNA。

Objective To explore the significance and the expression in transfected bone marrow mesenchymal cells by pcDNA3.1-BMP7. Methods pcDNA3.1-BMP7 was digested by enzymes and sequenced; pcDNA3.1-BMP7 was transfected into bone marrow mesenchymal cells of rabbit mediated by liposome and screened by G418 for 14 days. Expression of bone morphogenetic protein7 gene was examined with Western blot, PCR and in-situ hybridization. Results The bone morphogenetic protein7 gene pcDNA3.1-BMP7 was the same as the bone morpbogenetic protein7 gene reported. Massive expression of bone morpbogenetic protein7 and its mRNA was detected in cytoplasm of pcDNA3.1-BMP7 infected bone marrow mesenchymal cells with Western blot, PCR and insitu hybridization. Conclusion Mediated by liposome reagent, the bone marrow mesenchymal cells of rabbit are successfully transfected with pcDNA3.1-BMP7, and express bone morphogenetic protein7 gene.