flashBAC™ DNA Features: flashBAC™ DNA contains a bacterial artificial chromosome (BAC) that enables manufacturing of recombinant virus DNA in E. coli. lef2 and ORF1629 sequences flanking the BAC (in the flashBAC™ DNA) or gene of interest (in the transfer vector) allow for efficient homologous recombination in insect cells. A deletion in ORF1629 prevents replication of parental virus in insect cells. Deletion of the chitinase gene (chiA) maximizes production of secreted proteins and membrane-targeted proteins in insect cells.
Construction of Recombinant Virus Using flashBAC™: For baculovirus production, flashBAC™ DNA is mixed with transfer vector plasmid DNA containing the gene of interest to be inserted into the virus genome. Following co-transfection into insect cells, homologous recombination simultaneously removes the BAC, inserts the gene of interest and and restores ORF1629 allowing the production of infectious virus which can be harvested from the culture medium. Recombinant baculovirus can be used for subsequent transduction of insect or mammalian cells for protein expression.