骨形成蛋白2重组腺病毒的构建与鉴定  

Construction and identification of recombinant adenoviral vector-carrying BMP2

作　　者：王德利[1] 阮狄克[1] 李海峰[1] 马巍[2] 刘碧波[2] 杨敏杰[2] 

机构地区：[1]海军总医院骨外科,北京100037 [2]西安交通大学第一医院骨科

出　　处：《中华实验外科杂志》2004年第6期737-738,共2页Chinese Journal of Experimental Surgery

摘　　要：目的　通过构建骨形成蛋白 (BMP)重组腺病毒 ,为骨缺损的基因治疗提供可能。方法　应用逆转录 聚合酶链反应 (RT PCR)法克隆出BMP2全长基因 ,构建重组腺病毒载体 ,DNA 磷酸钙共沉法将辅助质粒PJM 17转染 2 93细胞 ,同源重组构建出重组腺病毒。测滴度并用氯化铯梯度离心纯化备用。结果　逆转录克隆出BMP2全长基因 1.2kb ,测序后连接于重组腺病毒载体 ,酶切鉴定后转染构建活病毒 ,再次感染 2 93细胞扩增 ,测滴度为× 10E10 pfu/ml ,并应用PCR法测定目的序列。Objective To construct a replication deficient type 5 adenoviral vector carrying BMP2 gene,AdCMVBMP2,for the treatment bone defect.Methods BMP2 gene was cloned by the RT PCR method to construct the recombinant adenoviral vector,and co transfected cell line 293 with helper plasmid PJM17,then the recombinant adenovirus was constructed by homologous recombination in the cell.Results The length of BMP2 gene was 1.2kb,and it were ligated to adenoviral vector to construct recombinant adenovirus.After measuring the titre of virus,the target gene was evaluated by PCR.Conclusion The successful construction of AdCMVBMP2 may provide an opportunity for the gene therapy of bone defect.

关 键 词：骨形成蛋白 基因治疗 腺病毒 RT-PCR 

分 类 号：R346[医药卫生—基础医学]