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Invitrogen™ Bac-to-Bac™ N-His TOPO™ Expression System Non-distribution product as customer accommodation.

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

Manufacturer:  Invitrogen™ A11101

Catalog No. A11101

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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.
  • Specifications


    Bac-to-Bac™ N-His TOPO™ Expression System, 20rxns, Liquid, Ampicillin (AmpR), Gentamicin (GmR) Antibiotic Resistance, Blunt TOPO™ Cloning Method, Baculovirus Expression, Cell-Based Expression Mechanism, Polyhedrin Promoter, His Tag Fusion, Cell-Based Expression Mechanism, Used for Protein Expression
    20 rxn(s)
    Protein expression, viral vaccine production, protein biology, protein expression, insect expression, high-level protein production in insect cells, baculovirus production and titering, bioproduction, viral vaccine production
    –30 to –10°C
    Bac-to-Bac™ N-His TOPO™ Expression System
    Vector kit for 20 topo cloning reactions, pFastBac/HBM-TOPO™ Vector containing the C-terminal TEV cleavage sit and His-Tag, pFastBac/CT-Gus control plasmid (Control expression vector), Other reagents supplied: 10X PCR, dNTP mix, salt solution, sterile water, control PCR template, polyhedrin forward primer, SV40 pA reverse primer, Kit containing competent cells (20 reactions), One Shot™ Mach1-T1R chemically competent E. coli, 4 kits with 5 x 0.1mL each of MAX Efficiency™ DH10Bac™ competent E. coli for 20 reactions, 1mL Cellfectin™ II reagent, Vector kit, One shot™ Mach1-T1R chemically competent E. coli, MAX Efficiency™ DH10Bac™ competent E. coli, Cellfectin™ II Reagent: store at 4°C
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