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Nucleic Acids Research, 2003, Vol. 31, No. 17 e101
© 2003 Oxford University Press

Improved generation of recombinant baculovirus genomes in Escherichia coli

Kari J. Airenne1, Erik Peltomaa1, Vesa P. Hytönen3, Olli H. Laitinen1 and Seppo Ylä-Herttuala*,1,2

1 AI Virtanen Institute, Department of Molecular Medicine and Biotechnology and 2 Department of Medicine and Gene Therapy Unit, University of Kuopio and Kuopio University Hospital, Kuopio, Finland and 3 Department of Biological and Environmental Science, University of Jyväskylä, Jyväskylä, Finland

*To whom correspondence should be addressed at AI Virtanen Institute, University of Kuopio, PO Box 1627, FIN-70211 Kuopio, Finland. Tel: +358 17 162076; Fax: +358 17 163751; Email: seppo.ylaherttuala{at}uku.fi
The authors wish it to be known that, in their opinion, the first two authors should be regarded as joint First Authors

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 ~107/µg 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.


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