A simplified system for generating recombinant adenoviruses

TC He, S Zhou, LT Da Costa, J Yu… - Proceedings of the …, 1998 - National Acad Sciences
Proceedings of the National Academy of Sciences, 1998National Acad Sciences
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 …
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.
National Acad Sciences