A protocol for rapid generation of recombinant adenoviruses using the AdEasy system.
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.
Luo
Jinyong
J
Key Laboratory of Diagnostic Medicine designated by the Ministry of Education, Chongqing Medical University, Chongqing 400046, China.
Deng
Zhong-Liang
ZL
Luo
Xiaoji
X
Tang
Ni
N
Song
Wen-Xin
WX
Chen
Jin
J
Sharff
Katie A
KA
Luu
Hue H
HH
Haydon
Rex C
RC
Kinzler
Kenneth W
KW
Vogelstein
Bert
B
He
Tong-Chuan
TC
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*
D049452:Green Fluorescent Proteins