Invitrogen™ Bac-to-Bac™ N-His TOPO™ Expression System

Description

• Bac-to-Bac™ Baculovirus Expression is an efficient method for producing baculovirus for high-level protein expression in insect cells
• It relies on generation of recombinant virus by site-specific transposition in E
• coli rather than homologous recombination in insect cells
• New: pFastBacTOPO™ vectors! Reliable and fast protein expression The Bac-to-Bac™ Baculovirus Expression System is faster and easier than traditional baculovirus expression, and it maintains high levels of protein expression
• Bac-to-Bac™ relies on generation of recombinant baculovirus by site-specific transposition in E
• coli rather than homologous recombination in insect cells
• The Bac-to-Bac™ Baculvirus Expression System is highly regarded in academic Literature
• Approximately 100 citations every year since 2000! Traditional baculovirus systems require purification and amplification of an initial low-titer viral supernatant
• This requires a time-consuming plaque assay
• Bac-to-Bac™'s pFastBac™ vector, however, recombines with the parent bacmid in DH10Bac™ E
• coli competent cells to form an expression bacmid
• Transfect the bacmid into insect cells for fast production of a high titer of pure recombinant baculovirus particles in the very first transfection
• You'll save weeks of precious time
• Collect a pure P2 baculovirus stock on Day 10 without the necessity of tedious plaques assay (See Figure 1) High expression, easy screening pFastBac™ uses the strong polyhedrin promoter to generate high levels of expression in a variety of insect cell line such as Sf9, Sf21, and High Five™ cells
• New additions: Bac-to-Bac™'s pFastBac vectors are now available as pFastBacTOPO™ vectors too
• pFastBacTOPO™ vectors are Blunt™ TOPO™ vectors and are supplied with Mach1™-T1R E
• coli for easy cloning and pFastBac⁄GOI propagation
• The new vectors enable you to: Amplify your gene of interest (GOI) with a PCR enzyme of highest fidelity such as Accuprime™ Pfx SuperMix
• TOPO™ clone the blunt-end PCR product into the new vector in only 5 minutes! Visualize colonies 8 hours after plating on ampicillin selective plates because Mach1™-T1R E
• coli cells have a faster doubling time compared to other standard cloning strains
• With the Bac-to-Bac™ N-His TOPO™ cloning or expression kit, you can produce a N-terminal His-fusion protein with a TEV cleavage site to purify with nickel-chelating resin (Invitrogen’s ProBond™ Purification System) and generate a native protein with the aid of Invitrogen’s popular AcTEV™ Protease
• The new vectors combine the TOPO™ cloning technology with the highly regarded Bac-to-Bac™ baculovirus expression features to enable easy cloning and high-level protein expression
• The pFastBacTOPO™ vectors are offered as cloning and expression kits.