中国冶金工业医学杂志

基因重组腺病毒载体AdFnCBD64构建的实验研究

作者：徐瑞剑王志强

分类号：R54

出版年,卷(期):页码：2015,32(3):249-251

摘要：

摘要：目的构建纤维连接蛋白羧基端细胞结合域（FnCBD64）基因重组人腺病毒载体（AdFnCBD64）。方法（1）FnCBD64重组腺病毒粘粒载体的构建；（2）293细胞和Hela细胞的培养；（3）磷酸钙转染；（4）FnCBD64基因重组腺病毒筛选和扩增；（5）设计腺病毒粘粒载体pAxCAwt公共PCR引物；（6）鉴定AdFnCBD64重组腺病毒；（7）浓缩与纯化重组腺病毒；（8）体外安全性研究。结果成功构建了纤维连接蛋白羧基端细胞结合域（FnCBD64）基因重组腺病毒载体AdFnCBD64，病毒滴度较高且安全性亦高。结论本实验成功构建了携带人FnCBD64基因的重组腺病毒，为应用FnCBD64促进骨髓间质干细胞黏附力提供了转染的载体，其使骨髓间质干细胞更快、更好地在体外构建组织工程心脏瓣膜成为可能。

关键词：腺病毒；基因重组；骨髓间质干细胞；组织工程；心脏瓣膜

Abstract:ObjectiveConstruction of carboxy\|terminal cell binding domain(FnCBD64)recombinant human adenovirus vector(AdFnCBD64)Methods1Construction of FnCBD64 adenovirus vector2Cultured 293 cells and Hela cells3Calcium phosphate transfection4Selected and amplified the adenovirus gene FnCBD645Designed the public PCR primers of adenovirus vector pAxCAwt6Identification AdFnCBD64 adenovirus vector7Concentrated and purified recombinant adenovirus vector8Studied safety in vitroResultsCarboxy\|terminal cell binding domain of fibronectin(FnCBD64)gene recombinant adenovirus vector AdFnCBD64 was successfully constructed Virus titers and the safety were highConclusionRecombinant adenovirus vector carrying human FnCBD64 is constructed successfully in this experiment,as transfected vectors which is provided for FnCBD64 promoting adhesion of bone mesenchymal stem cells(BMSCs)and make it possible that BMSCs construct in vitro tissue engineering heart valves more faster and betterKey Words:Adenovirus；Gene recombination；Mesenchymal stem cells；Tissue engineering；Heart valve

基金项目：

项目编号2009MS1148

作者简介：

参考文献：

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