A simplified system for generating recombinant adenoviruses

作者：Tong‐Chuan He, Shibin Zhou, Luís Teixeira da Costa, Jian Yu, Kenneth W. Kinzler, Bert Vogelstein · 发表于：Proceedings of the National Academy of Sciences · 年份：1998 · DOI：10.1073/pnas.95.5.2509 · 被引用次数：3242 · 研究领域：Virus-based gene therapy research、Viral Infectious Diseases and Gene Expression in Insects、CAR-T cell therapy research

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.