骨形成蛋白7重组腺病毒的构建及诱导成骨的研究  

Construction of human BMP7 recombinant adenovirus and its function of inducing bone formation

作　　者：孙大铭[1] 侯树勋[1] 付小兵[1] 

机构地区：[1]解放军总医院第一附属医院骨科,北京100037

出　　处：《中国骨肿瘤骨病》2008年第5期268-271,共4页Chinse Journal Of Bone Tumor And Bone Disease

基　　金：国家973课题项目(G1999054204);全军医学科研"十一五"计划课题(07A)

摘　　要：目的探讨骨形成蛋白7(BMP7)重组腺病毒异位诱导成骨的作用。方法将BMP7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BMP7重组腺病毒基因组,通过转染HEK293细胞包装出重组腺病毒,然后在裸鼠小腿肌肉中进行异位诱导成骨实验。结果经过PCR及酶切鉴定证明获得了BMP7转移质粒pAdTrack-BMP7和BMP7重组腺病毒基因组,并包装出重组腺病毒。组织学观察显示2周时实验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BMP7重组腺病毒的构建以及其异位诱导成骨的成功,为BMP7基因治疗的研究提供了确切的实验依据。Objective To construct and express a recombinant adenovirus carrying human BMP7 gene, and study its function of inducing bone formation. Methods BMP7 gene was cloned into the shuttle vectorpADTrack, and cotransfected into bacterium B J5183 together with the pADEasy vector to produce recombinant adenoviral plasmid by homologous recombination. Recombinant adenovirus was packaged in HEK293 cells. Then bone formation was studied in nude mice. Results PCR and digesting demonstrated that the shuttle plasmid-pADTrack-BMP7 and the recombinant adenoviral plasmid were obtained. The recombinant adenovirus was packaged in HEK293 cells. X-ray and histological methods showed new bone formation at 5W. Conclusions Construction and expression of human BMP7 recombinant adenovirus and its induction of new bone formation lay the basis for BMP7 gene therapy.

关 键 词：骨形成蛋白(BMP) 腺病毒 基因转染 

分 类 号：R68[医药卫生—骨科学]