A simplified system for generating recombinant adenoviruses

Proc Natl Acad Sci U S A. 1998 Mar 3;95(5):2509-14. doi: 10.1073/pnas.95.5.2509.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Adenoviridae / genetics*
  • Animals
  • Cell Line
  • Escherichia coli / genetics
  • Genes, Reporter
  • Genetic Vectors
  • Green Fluorescent Proteins
  • Humans
  • Kidney
  • Luminescent Proteins / biosynthesis
  • Mammals
  • Plasmids
  • Polymerase Chain Reaction
  • Recombinant Fusion Proteins / biosynthesis
  • Recombination, Genetic*
  • Repetitive Sequences, Nucleic Acid
  • Transfection / methods*

Substances

  • Luminescent Proteins
  • Recombinant Fusion Proteins
  • Green Fluorescent Proteins