人骨形态发生蛋白-7重组腺病毒载体的构建  

The construction of hbmp7 recombinant adenovirus vector

作　　者：李新友[1] 王金堂[1] 刘淼[1] 王全颖[1] 杨广笑[1] 李萌[1] 

机构地区：[1]西安交通大学医学院第一附属医院骨科,陕西西安710061

出　　处：《西安交通大学学报（医学版）》2008年第1期54-56,共3页Journal of Xi’an Jiaotong University（Medical Sciences）

摘　　要：目的为了研究人骨形态发生蛋白-7(hBMP7)导入骨缺损模型中的治疗作用,构建重组腺病毒载体。方法将hBMP7基因全长定向克隆入穿梭质粒pACCMV-plpA,构建pACCMV/rhBMP7穿梭质粒,应用磷酸钙-DNA共沉法将穿梭质粒及包装质粒PJM17共转染293细胞,经细胞内同源重组后,构建骨形态发生蛋白-7重组腺病毒Ad pAC-CMV/rhBMP7。结果成功构建了重组质粒pACCMV/rhBMP7,经293细胞包装后,扩增纯化测定病毒滴度为1×1011pfu/mL。结论成功构建人骨形态发生蛋白-7重组腺病毒Ad pACCMV/rhBMP7,为骨缺损、骨不连的基因治疗奠定了基础。Objective In order to investigate the treatment of bone defect repairing by recombinant adenovirus transudation the hBMP7 gene, the vector of hPMP7 recombinant Ad was constructed. Methods The gene of hBMP7 Was inserted into the replicated defective adenovirus shuttle plasmid pACCMV-plpA. We constructed the vector of hBMP7 recombinant Ad. The recombinant Ad viral stock was packaged. Totally 293 cells were co- transfected with the tAd vector of plasmid pACCMV/rhBMP7 and packaging plasmid PJM17 by calcium phosphate precipitation. Results The recombinant viral vector of plasmid pACCMV/rhBMP7 was constructed successfully. We got the Ad pACCMV/rhBMP7 stocks of high tire 1 × 10^11 pfu/mL. Conclusion We prepared the viral stock of rAd-BMP7 that can serve as the experimental study of gene therapy of bone defect repairing.

关 键 词：骨形态发生蛋白-7 腺病毒载体 基因转移 滴度测定 

分 类 号：Q78[生物学—分子生物学]