A protocol for rapid generation of recombinant adenoviruses using the AdEasy system
A protocol for rapid generation of recombinant adenoviruses using the AdEasy system
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DOI：
10.1038/nprot.2007.135
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发表时间：
2007-01-01
期刊：
影响因子：
14.8
通讯作者：
He, Tong-Chuan
中科院分区：
文献类型：
--
作者：
Luo, Jinyong;Deng, Zhong-Liang;He, Tong-Chuan
重组腺病毒为基因表达研究和治疗应用提供了一个通用的系统。我们已经开发出一种方法，可以简化这类病毒的生成和生产，称为AdEasy系统。用最少的酶操作产生重组腺病毒质粒，在细菌中而不是在真核细胞中使用同源重组。将这种载体导入哺乳动物包装细胞系后，在病毒骨架基因编码的绿色荧光蛋白的帮助下，病毒的生产就可以很方便地进行。该系统加速了用于各种目的的重组腺病毒的产生和测试过程。在这个方案中，我们描述了使用AdEasy系统产生重组腺病毒的实用方面。完整的方案通常需要4-5周的时间才能完成。
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.