title: Improved generation of recombinant baculovirus genomes in Escherichia coli.
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.
authors.affiliation: AI Virtanen Institute, Department of Molecular Medicine and Biotechnology, University of Kuopio and Kuopio University Hospital, Kuopio, Finland.
authors.forename: Kari J
authors.initials: KJ
authors.lastname: Airenne
authors.forename: Erik
authors.initials: E
authors.lastname: Peltomaa
authors.forename: Vesa P
authors.initials: VP
authors.lastname: Hytönen
authors.forename: Olli H
authors.initials: OH
authors.lastname: Laitinen
authors.forename: Seppo
authors.initials: S
authors.lastname: Ylä-Herttuala
mesh_terms: D001287:Attachment Sites, Microbiological / Q000235:genetics; D016367:Baculoviridae / Q000235:genetics*; D004251:DNA Transposable Elements / Q000235:genetics; D004279:DNA, Viral / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics*; D005822:Genetic Vectors / Q000235:genetics; D016679:Genome, Viral; D011995:Recombination, Genetic
chemical_list: D004251:DNA Transposable Elements; D004279:DNA, Viral