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. 2007;2(5):1236-47.
doi: 10.1038/nprot.2007.135.

A protocol for rapid generation of recombinant adenoviruses using the AdEasy system

Jinyong Luo  1 , Zhong-Liang Deng, Xiaoji Luo, Ni Tang, Wen-Xin Song, Jin Chen, Katie A Sharff, Hue H Luu, Rex C Haydon, Kenneth W Kinzler, Bert Vogelstein, Tong-Chuan He
Affiliations

A protocol for rapid generation of recombinant adenoviruses using the AdEasy system

Jinyong Luo et al. Nat Protoc. 2007.

Abstract

Recombinant adenoviruses provide a versatile system for gene expression studies and therapeutic applications. We have developed an approach that simplifies the generation and production of such viruses called the AdEasy system. A recombinant adenoviral plasmid is generated with a minimum of enzymatic manipulations, employing homologous recombination in bacteria rather than in eukaryotic cells. After transfection of such plasmids into a mammalian packaging cell line, viral production is conveniently followed with the aid of GFP encoded by a gene incorporated into the viral backbone. This system has expedited the process of generating and testing recombinant adenoviruses for a variety of purposes. In this protocol, we describe the practical aspects of using the AdEasy system for generating recombinant adenoviruses. The full protocol usually takes 4-5 weeks to complete.

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