Search
Citations & References (7)
Invitrogen™
pBAD/TOPO™ ThioFusion™ Expression Kit
The pBAD/TOPO™ ThioFusion™ Expression Kit is designed for one-step cloning and regulated expression of thioredoxin fusion proteins in E. coli(FigureRead more
| Catalog Number | Quantity |
|---|---|
| K37001 | 20 reactions |
Catalog number K37001
Price (USD)
1,406.00
Each
Quantity:
20 reactions
Price (USD)
1,406.00
Each
The pBAD/TOPO™ ThioFusion™ Expression Kit is designed for one-step cloning and regulated expression of thioredoxin fusion proteins in E. coli(Figure 1). The pBAD/Thio-TOPO™ vector encodes His-Patch thioredoxin as an N-terminal fusion partner. The thioredoxin fusion can significantly increase the solubility of many difficult-to-express proteins and improve the yield of protein production. The vector includes the following additional features (Figure 2):
• The araBAD promoter for tightly regulated expression in E. coli I-activated vector for 5-minute TOPO™ Cloning of Taq-amplified PCR products
• C-terminal polyhistidine (6xHis) tag for purification with nickel-chelating resin and detection with an Anti-His(C-term) Antibody
• C-terminal V5 epitope for detection with an Anti-V5 Antibody
• Enterokinase cleavage site for efficient cleavage of N-terminal fusion tag with EKMax™ Enterokinase
• The araBAD promoter for tightly regulated expression in E. coli I-activated vector for 5-minute TOPO™ Cloning of Taq-amplified PCR products
• C-terminal polyhistidine (6xHis) tag for purification with nickel-chelating resin and detection with an Anti-His(C-term) Antibody
• C-terminal V5 epitope for detection with an Anti-V5 Antibody
• Enterokinase cleavage site for efficient cleavage of N-terminal fusion tag with EKMax™ Enterokinase
For Research Use Only. Not for use in diagnostic procedures.
Specifications
Antibiotic Resistance BacterialAmpicillin (AmpR)
Bacterial or Yeast StrainTOP10
CleavageEK (Enterokinase) Recognition Site
Constitutive or Inducible SystemInducible
Expression MechanismCell-Based Expression
Expression SystemE. coli
Inducing AgentArabinose
Product TypeLentiviral Expression Kit
Quantity20 reactions
Selection Agent (Eukaryotic)None
VectorpBAD
Cloning MethodTOPO-TA
Product LineTOPO, ThioFusion
PromoteraraBAD
Protein TagHis Tag (6x), V5 Epitope Tag, Thioredoxin
Unit SizeEach
Contents & Storage
Each pBAD/TOPO™ ThioFusion™ Expression Kit contains two boxes and the LMG194 E. coli stab. The pBAD/TOPO™ ThioFusion™ Expression box contains all of the reagents necessary for PCR (except Taq polymerase) including 200 ng of topoisomerase I-activated pBAD/Thio-TOPO™ vector, sterile water, dNTPs, 10X PCR buffer, salt solution, control template and primers, 20% L-arabinose, primers for sequencing or PCR screening, and an expression control plasmid. Store at -20°C. The One Shot™ box contains all of the reagents necessary for transformation including 21 single-use 50 μl aliquots of chemically competent TOP10 E. coli, S.O.C. medium, and pUC19 supercoiled control plasmid. Store the One Shot™ box at -80°C. Store the LMG194 E. coli stab at 2 - 8°C. All components are guaranteed stable for 6 months when properly stored.
Frequently asked questions (FAQs)
How much L-arabinose should I use to induce expression with the pBAD expression system?
Should I use TOP10 cells or the LMG194 E. coli strain you offer for expression with my pBAD system?
What competent cells do you recommend I use for expression with my pBAD expression system?
Can you tell me the sequence of primers that can be used in pBAD vectors to sequence my gene of interest?
Do you have a list of the cap colors for the pBAD vectors?
Citations & References (7)
Citations & References
Abstract
A protein kinase encoded by the t complex responder gene causes non-mendelian inheritance
Journal:Nature
PubMed ID:10647005
'Males heterozygous for the t-haplotype form of mouse chromosome 17 preferentially transmit the t-chromosome to their progeny. Several distorter/sterility loci carried on the t-haplotype together impair flagellar function in all spermatozoa whereas the responder, Tcr, rescues t-sperm but not wild-type sperm. Thus, t-sperm have anadvantage over wild-type sperm in fertilizing
The cathepsin B of Toxoplasma gondii, toxopain-1, is critical for parasite invasion and rhoptry protein processing.
Journal:J Biol Chem
PubMed ID:12000756
'Cysteine proteinases play a major role in invasion and intracellular survival of a number of pathogenic parasites. We cloned a single copy gene, tgcp1, from Toxoplasma gondii and refolded recombinant enzyme to yield active proteinase. Substrate specificity of the enzyme and homology modeling identified the proteinase as a cathepsin B.
Bacterial cell killing mediated by topoisomerase I DNA cleavage activity.
Journal:J Biol Chem
PubMed ID:16159875
'DNA topoisomerases are important clinical targets for antibacterial and anticancer therapy. At least one type IA DNA topoisomerase can be found in every bacterium, making it a logical target for antibacterial agents that can convert the enzyme into poison by trapping its covalent complex with DNA. However, it has not
Degradation of corn fiber by Clostridium cellulovorans cellulases and hemicellulases and contribution of scaffolding protein CbpA.
Journal:Appl Environ Microbiol
PubMed ID:16000754
'Clostridium cellulovorans, an anaerobic bacterium, degrades native substrates efficiently by producing an extracellular enzyme complex called the cellulosome. All cellulosomal enzyme subunits contain dockerin domains that can bind to hydrophobic domains termed cohesins which are repeated nine times in CbpA, the nonenzymatic scaffolding protein of C. cellulovorans cellulosomes. In this
Factor B and the mitochondrial ATP synthase complex.
Journal:J Biol Chem
PubMed ID:11744738
Factor B is a subunit of the mammalian ATP synthase complex, whose existence has been controversial. This paper describes the molecular and functional properties of a recombinant human factor B, which when added to bovine submitochondrial particles depleted of their factor B restores the energy coupling activity of the ATP