Improving baculovirus recombination.
Recombinant baculoviruses have established themselves as a favoured technology for the high-level expression of recombinant proteins. The construction of recombinant viruses, however, is a time consuming step that restricts consideration of the technology for high throughput developments. Here we use a targeted gene knockout technology to inactivate an essential viral gene that lies adjacent to the locus used for recombination. Viral DNA prepared from the knockout fails to initiate an infection unless rescued by recombination with a baculovirus transfer vector. Modified viral DNA allows 100% recombinant virus formation, obviates the need for further virus purification and offers an efficient means of mass parallel recombinant formation.
Zhao
Yuguang
Y
Animal and Microbial Sciences, University of Reading, Whiteknights, Reading RG6 6AJ, UK.
Chapman
David A G
DA
Jones
Ian M
IM
D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics; D016367:Baculoviridae / Q000235:genetics*; D002460:Cell Line; D004279:DNA, Viral / Q000235:genetics; D004268:DNA-Binding Proteins; D004926:Escherichia coli / Q000235:genetics; D029968:Escherichia coli Proteins; D005090:Exodeoxyribonucleases / Q000235:genetics; D015870:Gene Expression; D005814:Genes, Viral / Q000235:genetics; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D009154:Mutation; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D011995:Recombination, Genetic; D014162:Transfection / Q000379:methods
D001426:Bacterial Proteins; D004279:DNA, Viral; D004268:DNA-Binding Proteins; D029968:Escherichia coli Proteins; D008164:Luminescent Proteins; C084497:RecT protein, E coli; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D005090:Exodeoxyribonucleases; C038225:recE protein, E coli