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Improved generation of recombinant baculovirus genomes in Escherichia coli
Kari J Airenne1, Erik Peltomaa, Vesa P Hytönen
1AI Virtanen Institute, Department of Molecular Medicine and Biotechnology, University of Kuopio and Kuopio University Hospital, Kuopio, Finland.
Abstract:
An improved method for the generation of recombinant baculoviruses by Tn7-mediated transposition is described. The method is based on the modified donor vector (pBVboost) and an improved selection scheme of the baculovirus bacmids in Escherichia coli with a mutated SacB gene. Recombinant bacmids can be generated at a frequency of approximately 10(7)/microg of donor vector with a negligible background. This easy-to-use and efficient pBVboost system provides the basis for a high-throughput generation of recombinant baculoviruses as well as a more convenient way to produce single viruses. The introduced selection scheme is also useful for the construction of other vectors by transposition in E.coli.