The Bac-to-Bac® Baculovirus Expression System is an efficient method for producing recombinant baculovirus for expression testing in insect cells. The Bac-to-Bac® System relies on generation of recombinant baculovirus by site-specific transposition in
E. coli rather than homologous recombination in insect cells (Figure 1). This system features:
- Time-saving expression bacmid. With Bac-to-Bac®, the expression cassette of the pFastBac™ vector recombines with the parent bacmid in DH10Bac™ E. coli Competent Cells to form an expression bacmid. The bacmid is then transfected into insect cells for production of recombinant baculovirus particles.
- Easy colony screening. The parent bacmid in DH10Bac™ E. coli contains a segment of the lacZα gene. The lacZα gene is disrupted upon transposition of the expression cassette into the bacmid allowing for blue/white selection of recombinants. This makes identification of recombinant colonies easy.
- Rapid purification of recombinant proteins. The pFastBac™ HT vector produces histidine-tagged recombinant protein. This allows rapid protein purification with nickel-chelating resin.
The Bac-to-Bac® Baculovirus Expression System is designed for fast, smallscale production of recombinant baculovirus. The pFastBac™ 1 vector offers the strong polyhedrin promoter for protein expression and a large multiple cloning site for simplified cloning.
