A simplified system for generating recombinant adenoviruses
A simplified system for generating recombinant adenoviruses
复制标题
DOI：
10.1073/pnas.95.5.2509
复制
发表时间：
1998-03-03
影响因子：
11.1
通讯作者：
Vogelstein, B
中科院分区：
文献类型：
--
作者：
He, TC;Zhou, SB;Vogelstein, B
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。我们在此报告了一种简化此类病毒的生成和生产的策略。重组腺病毒质粒是在细菌中而不是在真核细胞中使用同源重组以最少的酶促操作产生的。在将这些质粒转染到哺乳动物包装细胞系中后，借助于由掺入病毒骨架中的基因编码的绿色荧光蛋白，方便地进行病毒生产。从该程序中可以获得同质病毒而无需噬斑纯化。该系统将加快产生和测试用于各种目的的重组腺病毒的过程。
Recombinant adenoviruses provide a versatile system for gene expression studies and therapeutic applications. We report herein a strategy that simplifies the generation and production of such viruses. A recombinant adenoviral plasmid is generated with a minimum of enzymatic manipulations, using homologous recombination in bacteria rather than in eukaryotic cells. After transfections of such plasmids into a mammalian packaging cell line, viral production is conveniently followed with the aid of green fluorescent protein, encoded by a gene incorporated into the viral backbone. Homogeneous viruses can be obtained from this procedure without plaque purification. This system should expedite the process of generating and testing recombinant adenoviruses for a variety of purposes.