Invitrogen™ Bac-to-Bac™ N-His TOPO™ Cloning Kit
Bac-to-Bac™ N-His TOPO™ Cloning Kit
Supplier: Invitrogen™ A11099
DescriptionBac-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. It is based on 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. At least 80 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 lines such as Sf9, Sf21, and High Five™ cells.
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:
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.
|Ampicillin (AmpR), Gentamicin (GmR)|
|Kit content for Bac-to-Bac™ N-His TOPO™ cloning 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
Vector kit: store at -20°C
One Shot™ Mach1-T1R Chemically Competent E. coli: store at -80°C
|His Tag (6x)|
|High Five™, Sf21, Sf9|
|Insect Cell Expression|
|Protein Expression, Viral Vaccine Production|
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For Research Use Only. Not for use in diagnostic procedures.