CONSTRUCTION AND EXPRESSION OF ADENO-ASSOCIATED VIRUS-BASED PLASMID EXPRESSING VECTORS CONTAINING hIL-2 GENE OR mIFN-γ GENE

作     者：张景迎 梁宏立 陈诗书 ZHANG Jingying;LIANG Hongli;CHEN Shishu(Human Gene Therapy Research Center, SSMU Shanghai(200025))

作者机构：Human Gene Therapy Research Center SSMU Shanghai200025 

出 版 物：《Medical Bulletin of Shanghai Jiaotong University》 (上海交通大学学报（医学英文版）)

年 卷 期：2000年第21卷第1期

页      面：14-17页

学科分类：0710[理学-生物学] 07[理学] 071007[理学-遗传学] 

基　　金：国家自然科学基金(39730440) 

主　　题：adeno associated virus plasmid interleukin 2 interferon γ gene transfer 

摘      要：Obiective TO improve the plasmid vectors in gene therapy, adeno - associated virus (AAV) basedplasmid expressing vectors containing hIL - 2 gene or mIFN-γ gene were constructed and its expression intransfected cells was studied. Methods By means of step to step cloning, promoter CMVp was placed at thedownstream of 5’ inverted terminal repeat from AAV (AAV - ITR) of pAP, hIL - 2 gene or mIFN -γ gene insertedinto pAC between CMVp and polyA. Then intron A was inserted into pAC - hIL - 2 or pAC- mIFN-γ betweenCMVp and IL - 2 gene or IFNγ gene to construct pAI- hIL - 2 or pAI- mIFN -γ. Liposome - plasmid complexeswere formed by mixing Dosper with these AAV- based plasmids containing hIL - 2 gene or mIFN- γgene. Results High biotogical activities of IL - 2 or IFN- γ could be detected in the supernatants of NIH3T3 andMM45T Li cells after transfection. Insertion of intron A into pAC- hIL - 2 or pAC- mIFN - γ improved theexpression of IL - 2 or IFN- γ. Conclusion These data demonstrated that the constructed AAV-based plasmidexpressing vectors could ejlciently express therapeutic genes in cultured cells and could be used as a nonviral genetransfer system in human gene therapy.