The Bac-to-Bac® Vector Kit contains a pFastBac™ 1 vector, as well an expression control vector, intended for use as part of the Bac-to-Bac® Baculovirus Expression System (Cat. No. 10359-016), which enables the efficent production of 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 (not included with this vector kit) 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.
The Bac-to-Bac® Baculovirus Expression System is designed for fast, small-scale production of recombinant baculovirus. The pFastBac™ 1 vector provided by this kit offers the strong polyhedrin promoter for protein expression and a large multiple cloning site for simplified cloning.