Rapid Construction of EGFP Labled Recombinant Adenovirus Containing hVEGF165 and Its Expression in Haematopoietic Cells

作     者：仲照东 邹萍 黄士昂 胡中波 刘凌波 卢运萍 

作者机构：Institute of Haematology Xiehe Hospital Tongji Medical College Huazhong University of Science and Technology Biological Center of Cancer Tongji Hospital Tongji Medical College Huazhong University of Science and Technology 

出 版 物：《Journal of Huazhong University of Science and Technology(Medical Sciences)》 (华中科技大学学报（医学英德文版）)

年 卷 期：2003年第23卷第1期

页      面：4-6页

核心收录：

学科分类：0831[工学-生物医学工程（可授工学、理学、医学学位）] 0710[理学-生物学] 1001[医学-基础医学(可授医学、理学学位)] 0805[工学-材料科学与工程（可授工学、理学学位）] 0703[理学-化学] 0836[工学-生物工程] 10[医学] 

基　　金：This project was supported by the Overseas Scholar Cooper-ative Foundation ( No. 3 992 80 10 ) 

主　　题：EGFP vascular endothelial growth factor adenovirus homologous recombination haematopoietic cells 

摘      要：By using AdEasy system, which is based on the homologous recombination in bacteria, an EGFP labeled recombinant adenovirus vector containing hVEGF165 was constructed quickly and efficiently expressed in mouse haematopoietic cells. First, hVEGF165 coding sequence was subcloned into shuttle plasmid pAdTrack-CMV, then cotransformed with adenoviral backbone vector pAdEasy-1 into *** strain BJ5183. The recombinant adenoviral plasmid was transfected into HEK293 cells to assembly replication-defective adenovirus Ad-EGFP/hVEGF165. The expression of EGFP could be easily detected. The rate of EGFP positive mouse bone marrow mononuclear cells by flow cytometric analysis was 27.3 % (MOI=100), and the expression of hVEGF165 protein in the supernatant was (1385±332) pg/10 6 cells. These results suggest that the construction of adenovirus vector by homologous recombination in bacteria features high efficiency and simplicity. The prepared high titer Ad-EGFP/hVEGF165 can be used an efficient helpful vector to infect hematopoietic cells.