骨形成蛋白2重组腺病毒的构建及异位诱导成骨的研究

Construction of a recombinant adenovirus carrying human BMP2 gene and its function of inducing bone formation

目的构建骨形成蛋白2(BMP2)重组腺病毒,探讨其诱导成骨的作用。方法将BMP2基因克隆到转移载体pAdTrack-CMV中,在细菌BJ5183中与pAdEasy腺病毒基因组进行同源重组,得到BMP2重组腺病毒基因组,通过转染HEK293细胞,包装出重组腺病毒。然后,用其进行裸鼠异位诱导成骨。结果经过PCR及酶切鉴定,证明获得了BMP2转移质粒pAdTrack-BMP2和BMP2腺病毒基因组,并包装出重组腺病毒。组织学观察显示,2周时局部大量纤维样细胞聚集,软骨细胞分化;5周时骨小梁形成,软骨细胞已退化。结论BMP2组腺病毒的构建及其异位诱导成骨的功能为其在BMP2基因治疗中的应用奠定了良好的基础。

Objective To construct a recombinant adenovirus carrying human BMP2 gene, and study its function of inducing hone formation. Methods BMP2 gene was cloned into the shuttle vector - pADtrack ; the bacterium BJ5183 was transfected with the shuttle and the pADEasy vector and the recombinant adenoviral plasmid was produced by homologous recombination. Recombinant adenovirus was packaged in HEK293 cells. Then bone formation was studied in nude mice. Results PCR and endonucleas digesting demonstrated that the shuttle plasmidpADTrack - BMP2 and the recombinant adenoviral plasmid were successfully obtained. The recombinant adenovirus was packaged in HEK293 cells. Histological method showed new bone formation at 5W. Conclusion Construction of BMP2 recombinant adenovirus and its induction of new bone formation was the basis of BMP2 gene therapy.