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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 onRead more
| Catalog Number | Quantity |
|---|---|
| A11098 | 20 Reactions |
Catalog number A11098
Price (HKD)
16,448.00
Each
Quantity:
20 Reactions
Price (HKD)
16,448.00
Each
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 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.
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 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.
For Research Use Only. Not for use in diagnostic procedures.
Specifications
Antibiotic Resistance BacterialAmpicillin (AmpR), Gentamicin (GmR)
Cell LineHigh Five™, Sf21, Sf9
Cell TypeInsect Cell
Cloning MethodBlunt TOPO
Expression MechanismInsect Cell Expression
Expression SystemBaculovirus
For Use With (Application)Protein Expression, Viral Vaccine Production
No. of Reactions20 Reactions
Product LineBac-to-Bac, TOPO
Product TypeCloning Kit
PromoterPolyhedrin
Protein TagHis Tag (6x)
Quantity20 Reactions
Selection Agent (Eukaryotic)Ampicillin, Gentamicin
VectorpFastBac/CT-TOPO
Unit SizeEach
Contents & Storage
Kit content for Bac-to-Bac™ C-His TOPO™ cloning kit (For details see the manual)
• Vector kit for 20 topo cloning reactions:
o pFastBac⁄CT-TOPO™ Vector containing the C-terminal TEV cleavage sit and His-Tag.
o pFastBac⁄CT-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
• Vector kit for 20 topo cloning reactions:
o pFastBac⁄CT-TOPO™ Vector containing the C-terminal TEV cleavage sit and His-Tag.
o pFastBac⁄CT-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