Description
The original flashBAC™ vector (cat# 100150/100151/100152) is a good choice for projects where the gene to be expressed is likely to be simple and targeted to the cytoplasm or nucleus of infected cells. The flashBAC ™ system builds on the BacPAK6 technology. At the heart of the new system is an AcMNPV genome that lacks part of the essential gene ORF 1629 and contains a bacterial artificial chromosome (BAC) at the pohl locus, replacing the polh coding sequence. The essential gene deletion prevents virus replication in insect cells and the BAC allows the virus genome to be maintained in bacterial cells as a bacmid. A recombinant baculovirus is produced simply by co-transfecting insect cells with flashBAC ™ DNA and a compatible transfer plasmid containing the gene to be expressed. Homologous recombination within the insect cells removes the BAC sequences, inserts the foreign gene under control of the polh promoter, and restores ORF 1629 allowing the recombinant virus to replicate.
Using the flashBAC™ system is easier and quicker than other baculovirus expression systems because there is no need to separate recombinant virus from parental virus by plaque-purification or any other means; only recombinant virus is produced after the co-transfection. Because the production of recombinant virus has been reduced to a single step procedure in insect cells, it is amenable to high throughput and automated production systems. However, it is also of benefit to the small research group requiring a low cost solution to producing one or a few recombinant baculoviruses prepared in individual dishes of cells.
