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

Catalog number: A11101

Invitrogen™  Related applications: Protein Expression | Viral Vaccine Production

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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.
For Research Use Only. Not for use in diagnostic procedures.


Promoter: Polyhedrin
Expression: Baculovirus
Product Size: 20 reactions
Cloning Method: Blunt TOPO®
Expression Mechanism: Cell-Based Expression
Protein Tag or Fusion: His Tag (6x)
Antibiotic Resistance (Bacterial): Ampicillin (AmpR), Gentamicin (GmR)

Contents & storage

Kit content for Bac-to-Bac® N-His TOPO® Expression kit (For details see the manual)
• Vector Kit for 20 topo cloning reactions:
o pFastBac⁄NT-TOPO® Vector containing the N-terminal His-Tag and TEV cleavage site.
o pFastBac⁄NT-Gus Control Plasmid (Control expression vector)
o 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):
o One Shot® Mach1-T1R Chemically Competent E. coli
• MAX Efficiency® DH10Bac™ Competent E. coli for 20 reactions (4 kits with 5x 0.1 ml each)
• 1 ml Cellfectin® II Reagent
Vector kit: store at -20°C
One Shot® Mach1-T1R Chemically Competent E. coli: store at -80°C
MAX Efficiency® DH10Bac™ Competent E. coli: store at -80°C
Cellfectin® II Reagent: store at +4°C


Manuals & protocols