Improved generation of recombinant baculovirus genomes in Escherichia coli.
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.
Airenne
Kari J
KJ
AI Virtanen Institute, Department of Molecular Medicine and Biotechnology, University of Kuopio and Kuopio University Hospital, Kuopio, Finland.
Peltomaa
Erik
E
Hytönen
Vesa P
VP
Laitinen
Olli H
OH
Ylä-Herttuala
Seppo
S
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
D004251:DNA Transposable Elements; D004279:DNA, Viral