Author:
Zhou Xiangyang,Sena-Esteves Miguel,Gao Guangping
Abstract
This protocol describes how to generate an infectious adenovirus vector by direct ligation and cloning. This is the first step in the production of a recombinant adenovirus vector. The protocol begins with a convenient and efficient double-selection procedure based on antibiotic resistance and identification of green fluorescent protein (GFP)-negative bacterial colonies. In this protocol, the prokaryotic expression cassette for GFP in the shuttle plasmid pSh-pkGFP is replaced with the transgene. The resulting pShuttle–transgene plasmid is then used to clone the transgene expression cassette into pAd-pkGFP, using the same double-selection procedure.
Publisher
Cold Spring Harbor Laboratory
Subject
General Biochemistry, Genetics and Molecular Biology
Cited by
5 articles.
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