Thermo Fisher, A11098, Bac-to-Bac™ C-His TOPO™ Cloning Kit

Bac-to-Bac™ C-His TOPO™ Cloning Kit 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 expressionThe 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 screeningpFastBac™ uses the strong polyhedrin promoter to generate high levels of expression in a variety of insect cell lines 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™ C-His TOPO™ cloning or expression kit, you can produce a C-terminal His-fusion protein with a TEV cleavage site to purify with nickel-chelating resins (Invitrogen’s ProBond™ Purification System) and generate a native protein with the aid of Invitrogen’s popular AcTEV™ Protease. This is the Invitrogen’s only vector that contains a C-terminal TEV cleavage site!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. Antibiotic Resistance Bacterial: Ampicillin (AmpR), Gentamicin (GmR)
Cell Line: High Five™, Sf21, Sf9
Cell Type: Insect Cell
Cloning Method: Blunt TOPO
Expression Mechanism: Insect Cell Expression
Expression System: Baculovirus
For Use With (Application): Protein Expression, Viral Vaccine Production
No. of Reactions: 20 Reactions
Product Line: Bac-to-Bac™, TOPO™
Product Type: Cloning Kit
Promoter: Polyhedrin
Protein Tag: His Tag (6x)
Quantity: 20 Reactions
Selection Agent (Eukaryotic): Ampicillin, Gentamicin
Vector: pFastBac/CT-TOPO
Unit Size: Each