Nature Methods2, 95 - 97 (2005)
Published online: 21 January 2005; | doi:10.1038/nmeth734
Engineering of a vaccinia virus bacterial artificial chromosome in Escherichia coli by bacteriophage −based recombination
Arban Domi
& Bernard Moss
Laboratory of Viral Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, 4 Center Drive, Bethesda, Maryland 20892-0445, USA.
Correspondence should be addressed to Bernard Moss bmoss@nih.gov
The large capacity of vaccinia virus (VAC) for added DNA, cytoplasmic expression and broad host range make it a popular choice for gene delivery, despite the burdensome need for multiple plaque purifications to isolate recombinants. Here we describe how a bacterial artificial chromosome (BAC) containing the entire VAC genome can be engineered in Escherichia coli by homologous recombination using bacteriophage −encoded enzymes. The engineered VAC genomes can then be used to produce clonally pure recombinant viruses in mammalian cells without the need for plaque purification.
MORE ARTICLES LIKE THIS
These links to content published by NPG are automatically generated.
−based recombination
−encoded enzymes. The engineered VAC genomes can then be used to produce clonally pure recombinant viruses in mammalian cells without the need for plaque purification.
