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

被引：713

作者：

Luo, Jinyong ^{[1
,2
]}

Deng, Zhong-Liang ^{[2
,3
]}

Luo, Xiaoji ^{[1
,2
]}

Tang, Ni ^{[1
,2
]}

Song, Wen-Xin ^{[2
]}

Chen, Jin ^{[1
,2
]}

Sharff, Katie A. ^{[2
]}

Luu, Hue H. ^{[2
]}

Haydon, Rex C. ^{[2
]}

Kinzler, Kenneth W. ^{[4
,5
]}

Vogelstein, Bert ^{[4
,5
]}

He, Tong-Chuan ^{[1
,2
]}

机构：

[1] Chongqing Med Univ, Minist Educ, Key Lab Diag Med, Chongqing 400046, Peoples R China

[2] Univ Chicago, Med Ctr, Dept Surg, Mol Oncol Lab, Chicago, IL 60637 USA

[3] Chongqing Med Univ, Affiliated Hosp 2, Dept Orthopaed Surg, Chongqing, Peoples R China

[4] Kimmel Comprehens Canc Ctr Johns Hopkins, Howard Hughes Med Inst, Baltimore, MD 21231 USA

[5] Ludwig Ctr Canc Genet & Therapeut, Kimmel Comprehens Canc Ctr Johns Hopkins, Baltimore, MD 21231 USA

来源：

NATURE PROTOCOLS | 2007年 / 2卷 / 05期

基金：

中国国家自然科学基金; 美国国家卫生研究院;

关键词：

D O I：

10.1038/nprot.2007.135

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

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.

引用

页码：1236 / 1247

页数：12

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