flashBAC™ ULTRA Baculovirus Ex

INTRODUCTION
The flashBAC™ System is a streamlined platform for the production of recombinant baculoviruses. Deletion of the essential gene ORF1629 from the Autographa californica nucleopolyhedrovirus
(AcMNPV) genome prevents non-recombinant virus from replicating within insect cells, thus eliminating the need to plaque-purify recombinant virus from parental virus. Secreted or membranetargeted recombinant protein yields are greatly increased by the deletion of the chitinase gene (chiA). Further nonessential gene deletions in flashBAC™ ULTRA provide enhanced quality and yield for difficult to express proteins. The insertion of a bacterial artificial chromosome (BAC) at the AcMNPV polyhedrin gene locus allows viral DNA to be maintained and propagated as a circular genome within bacterial cells. The genomic DNA can then be purified and is the flashBAC™ DNA provided with this kit.
To generate recombinant baculovirus using the flashBAC™ System, insect cells are transfected with  TransIT®-Insect Transfection Reagent, flashBAC™ DNA, and a transfer plasmid (e.g. pOET1, pOET1C_6xHIS, or pOET6) containing the gene of interest. Homologous recombination within the insect cells (see schematic at right) inserts the gene of interest under the control of the polyhedrin promoter, and restores the function of ORF1629 allowing viral DNA to replicate and produce virus particles. The recombinant virus genome replicates to produce baculovirus that can be harvested directly from the culture medium of transfected insect cells. Because the flashBAC™ System has effectively reduced recombinant baculovirus production to a one-step procedure, it is fully amenable to high throughput and automated production platforms. More details on the flashBAC™ and flashBAC™ ULTRA Systems are available at www.mirusbio.com/flashbac.