The flashBAC system is a streamlined platform designed for the production of recombinant baculoviruses

Further nonessential gene deletions in flashBAC GOLD and 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 (pOET1, pOET1C_6xHIS, or pOET6) containing the gene of interest.

Homologous recombination within the insect cells 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.

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).