title: Optimizing the baculovirus expression vector system.
abstract: Baculoviruses have a unique bi-phasic life cycle and powerful promoters, which greatly facilitates their use for recombinant protein expression in insect cells. We have developed an expression system that utilizes homologous recombination in insect cells between a transfer plasmid containing a gene to be expressed and a replication-deficient virus (bacmid). Only recombinant virus can replicate facilitating the rapid production of multiple recombinant viruses using robotic liquid handlers. The bacmid has also been genetically optimized for improved protein expression and stability. We describe the application of this system for high level production of recombinant proteins.
authors.affiliation: Oxford Expression Technologies Ltd., Oxford OX3 0BP, United Kingdom. r.hitchman@oetltd.com
authors.forename: Richard B
authors.initials: RB
authors.lastname: Hitchman
authors.forename: Elisabetta
authors.initials: E
authors.lastname: Locanto
authors.forename: Robert D
authors.initials: RD
authors.lastname: Possee
authors.forename: Linda A
authors.initials: LA
authors.lastname: King
mesh_terms: D000818:Animals; D057205:Automation, Laboratory; D016367:Baculoviridae / Q000235:genetics*; D002460:Cell Line; D015870:Gene Expression; D005822:Genetic Vectors / Q000737:chemistry / Q000235:genetics*; D057166:High-Throughput Screening Assays; D059765:Homologous Recombination; D010957:Plasmids / Q000737:chemistry / Q000235:genetics*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D018411:Spodoptera / Q000166:cytology / Q000378:metabolism* / Q000821:virology; D014779:Virus Replication / Q000235:genetics
chemical_list: D011994:Recombinant Proteins