A simplified system for generating recombinant E3-deleted canine adenovirus-2

文献类型： 外文期刊

第一作者： Yu, Zuo

作者： Yu, Zuo;Jiang, Qian;Liu, Jiasen;Guo, Dongchun;Quan, Chuansong;Li, Botao;Qu, Liandong

作者机构：

关键词： Canine adenovirus type 2;Vector;Homologous recombination;Shuttle vector;Enhanced green fluorescent protein

期刊名称：PLASMID （ 影响因子：3.466； 五年影响因子：2.74 ）

ISSN： 0147-619X

年卷期： 2015 年 77 卷

页码：

收录情况： SCI

摘要： Canine adenovirus type 2 (CAV-2) has been used extensively as a vector for studying gene therapy and vaccine applications. We describe a simple strategy for generating a replication-competent recombinant CAV-2 using a backbone vector and a shuttle vector. The backbone plasmid containing the full-length CAV-2 genome was constructed by homologous recombination in Escherichia coli strain BJ5183. The shuttle plasmid, which has a deletion of 1478 bp in the nonessential E3 viral genome region, was generated by subcloning a fusion fragment containing the flanking sequences of the CAV-2 E3 region and expression cassette sequences from pcDNA3.1(+) into modified pUC18. To determine system effectiveness, a gene for enhanced green fluorescent protein (EGFP) was inserted into the shuttle plasmid and cloned into the backbone plasmid using two unique NruI and SalI sites. Transfection of Madin-Darby canine kidney (MDCK) cells with the recombinant adenovirus genome containing the EGFP expression cassette resulted in infectious viral particles. This strategy provides a solid foundation for developing candidate vaccines using CAV-2 as a delivery vector. (C) 2014 Published by Elsevier Inc.

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