摘要
目的 构建骨形成蛋白 (BMP) 7重组腺病毒 ,使其转染骨髓间充质干细胞并在细胞内得到表达。方法 将BMP7基因克隆到转移载体 pAdTrack CMV中 ,在细菌BJ5 183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HEK2 93细胞包装出重组腺病毒。利用BMP7重组腺病毒转染分离纯化后的骨髓间充质干细胞 ,鉴定BMP7在细胞内的表达。结果 经过多聚酶链反应 (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。逆转录PCR和免疫组织化学证明BMP7在重组腺病毒转染后的骨髓间充质干细胞中得到了表达。结论 BMP7重组腺病毒的构建和在骨髓间充质干细胞中的表达 ,为BMP7基因治疗的研究奠定了基础。
Objective To construct a recombinant adenovirus carrying human BMP7 gene which is transfected into marrow mesenchymal stem cells (MSCs) and expressed.Methods BMP7 gene was cloned into the shuttle vector pADTrack; the bacterium BJ5183 was contransfected with the shuttle and the pADEasy vector and the recombinant adenoviral plasmid was produced by homologous.Recombinant adenovirus was packaged in HEK293 cells.Marrow MSCs were transfected with recombinant adenovirus and BMP7 expression was confirmed.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.RT PCR and immunohistochemical methods revealed that BMP7 was expressed in MSCs.Conclusion Construction of human BMP7 recombinant adenovirus and its expression in marrow MSCs laid the basis for BMP7 gene therapy.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2003年第5期436-438,共3页
Chinese Journal of Experimental Surgery
基金
国家 973计划资助项目 (G1 9990 542 0 4 )
关键词
骨形成蛋白7
腺病毒
构建
转染
骨髓问充质干细胞
Bone morphogenetic protein
Adenovirus
Marrow mesenchymal stem cells
Gene transfection
