flashBAC Baculovirus Expression System – 5 Reactions

Description

flashBAC™ Baculovirus Expression Systems are a streamlined platform for the production of recombinant baculoviruses. Genetic optimization of the Autographa californica nucleopolyhedrovirus (AcMNPV) genome yields recombinant virus in a single step and eliminates the need for separation of recombinant and parental virus by plaque purification. flashBAC™ systems and pOET Insect and BacMam Transfer Plasmids are optimized for use with TransIT®-Insect Transfection Reagent.

Benefits of the flashBAC™ system include:

Ease-of-Use – Single-step baculovirus generation in insect cells, no plaque purification required
Compatibility – Ready to use with most transfer vectors (PDF), including BacMam
More Protein – High recombinant protein yields

SKU: MIR 6115

Supporting Data

flashBAC™ Systems offered by Mirus Bio:

System	Description
flashBAC™	The original flashBAC™ expression vector. Protein expression and secretion is improved by deletion of the chitinase gene from the AcMNPV genome. Optimal for expression of most nuclear or cytoplasmic proteins.
flashBAC™ ULTRA	The most advanced flashBAC™ vector system, optimized by deletions in the chitinase, cathepsin, p10, p74 and p26 genes. The ideal system for the most difficult to express proteins including nuclear, cytoplasmic, membrane and secreted proteins.
Additional details on the genetic modifications included in the flashBAC™ systems are described in Hitchman et al. 2010.

How does the flashBAC™ Baculorvirus Expression System work?

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.

flashBAC™ Systems and pOET vectors are sold by Mirus Bio through partnership with Oxford Expression Technologies, Oxford, UK.