He Tong-Chuan
The University of Chicago Medical Center, Chicago, Illinois, USA.
Curr Protoc Hum Genet. 2004 May;Chapter 12:Unit 12.4. doi: 10.1002/0471142905.hg1204s40.
Adenoviral vectors have been widely used as efficient gene delivery vehicles for gene therapy and vaccine development, as well as gene function studies. The recently developed AdEasy system is a simple and efficient method for rapid generation of recombinant adenoviruses. Unlike traditional adenoviral vectors, the viral backbone described here is supplied as a supercoiled plasmid rather than as a linear viral DNA, facilitating backbone amplification. The recombination step is performed in E. coli rather than in mammalian cells, taking advantage of the high efficiency of homologous recombination in bacteria. This unit includes the complete set of protocols needed to generate recombinant adenoviruses using the AdEasy system.
腺病毒载体已被广泛用作基因治疗、疫苗开发以及基因功能研究的高效基因递送载体。最近开发的AdEasy系统是一种快速产生重组腺病毒的简单高效方法。与传统腺病毒载体不同,这里描述的病毒骨架以超螺旋质粒形式提供,而不是线性病毒DNA,便于骨架扩增。重组步骤在大肠杆菌中进行,而不是在哺乳动物细胞中进行,利用了细菌中同源重组的高效率。本单元包括使用AdEasy系统产生重组腺病毒所需的完整协议集。