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

A simplified system for generating recombinant adenoviruses.

Author information

  • 1The Howard Hughes Medical Institute, 424 North Bond Street, Baltimore, MD 21231, USA. tche@welchlink.welch.jhu.edu

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.

PMID:
9482916
[PubMed - indexed for MEDLINE]
PMCID:
PMC19394
Free PMC Article

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