title: A protocol for rapid generation of recombinant adenoviruses using the AdEasy system.
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.
authors.affiliation: Key Laboratory of Diagnostic Medicine designated by the Ministry of Education, Chongqing Medical University, Chongqing 400046, China.
authors.forename: Jinyong
authors.initials: J
authors.lastname: Luo
authors.forename: Zhong-Liang
authors.initials: ZL
authors.lastname: Deng
authors.forename: Xiaoji
authors.initials: X
authors.lastname: Luo
authors.forename: Ni
authors.initials: N
authors.lastname: Tang
authors.forename: Wen-Xin
authors.initials: WX
authors.lastname: Song
authors.forename: Jin
authors.initials: J
authors.lastname: Chen
authors.forename: Katie A
authors.initials: KA
authors.lastname: Sharff
authors.forename: Hue H
authors.initials: HH
authors.lastname: Luu
authors.forename: Rex C
authors.initials: RC
authors.lastname: Haydon
authors.forename: Kenneth W
authors.initials: KW
authors.lastname: Kinzler
authors.forename: Bert
authors.initials: B
authors.lastname: Vogelstein
authors.forename: Tong-Chuan
authors.initials: TC
authors.lastname: He
mesh_terms: D000256:Adenoviridae / Q000235:genetics*; D001419:Bacteria; D002478:Cells, Cultured; D005818:Genetic Engineering / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics*; D049452:Green Fluorescent Proteins / Q000235:genetics; D006801:Humans; D011995:Recombination, Genetic; D014162:Transfection / Q000379:methods*
chemical_list: D049452:Green Fluorescent Proteins