pBAD TOPO™ TA Expression Kit
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Citations & References (13)
Invitrogen™

pBAD TOPO™ TA Expression Kit

The pBAD TOPO™ TA Expression Kit is specifically designed for one-step cloning and regulated prokaryotic expression of Taq-amplified PCR products.Read more
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Catalog NumberProtein TagQuantity
K430001His Tag (6x), V5 Epitope Tag20 Reactions
K430040V5 Epitope Tag40 Reactions
Catalog number K430001
Price (CNY)
13,572.00
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Ends: 31-Dec-2026
15,873.00
Save 2,301.00 (14%)
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Protein Tag:
His Tag (6x), V5 Epitope Tag
Quantity:
20 Reactions
Price (CNY)
13,572.00
飞享价
Ends: 31-Dec-2026
15,873.00
Save 2,301.00 (14%)
Each
Add to cart
The pBAD TOPO™ TA Expression Kit is specifically designed for one-step cloning and regulated prokaryotic expression of Taq-amplified PCR products. Once you've TOPO™ Cloned your PCR product, you can go straight to protein expression. Some of the convenient features of the pBAD-TOPO™ vector include:

• Linearized, topoisomerase I-activated vector for 5-minute cloning of Taqamplified PCR products
• The araBAD promoter for tightly regulated expression in E. coli V5 epitope tag for detection with an Anti-V5 Antibody
• C-terminal polyhistidine (6xHis) tag for purification using nickel-chelating resin and detection with an Anti-His(C-term) Antibody
• Enterokinase cleavage site for removal of N-terminal leader peptide
For Research Use Only. Not for use in diagnostic procedures.
Specifications
Antibiotic Resistance BacterialAmpicillin (AmpR)
Bacterial or Yeast StrainLMG194, TOP10
CleavageEK (Enterokinase) Recognition Site
Constitutive or Inducible SystemInducible
Expression MechanismCell-Based Expression
Expression SystemE. coli
Inducing AgentArabinose
Product TypeTA Expression Kit
Quantity20 Reactions
Selection Agent (Eukaryotic)None
VectorpBAD
Cloning MethodTOPO-TA
Product LineTOPO
PromoteraraBAD
Protein TagHis Tag (6x), V5 Epitope Tag
Unit SizeEach
Contents & Storage
Each pBAD TOPO™ TA Expression Kit contains two boxes and the LMG194 E. coli stab. The pBAD TOPO™ TA box contains all of the reagents necessary for PCR (except Taq polymerase), including 200 ng of topoisomerase I-activated pBAD-TOPO™ vector, sterile water, dNTPs, 10X PCR buffer, salt solution, control template and primers, 20% L-arabinose, pBAD 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 single-use 50 μl aliquots of One Shot™ TOP10 Chemically Competent E. coli, S.O.C. medium, and a supercoiled control plasmid. Store One Shot™ cells at -80°C. Store the LMG194 E. coli stab at 2 - 8°C. All reagents are guaranteed stable for 6 months when properly stored.

Frequently asked questions (FAQs)

Can I store my competent E. coli in liquid nitrogen?

We do not recommend storing competent E. coli strains in liquid nitrogen as the extreme temperature can be harmful to the cells. Also, the plastic storage vials are not intended to withstand the extreme temperature and may crack or break.

How should I store my competent E. coli?

We recommend storing our competent E. coli strains at -80°C. Storage at warmer temperatures, even for a brief period of time, will significantly decrease transformation efficiency.

How much L-arabinose should I use to induce expression with the pBAD expression system?

While the amount of L-arabinose can vary depending on your expression experiment, we suggest performing a pilot expression experiment with varying amounts of L-arabinose from 0.00002% to 0.2%.

Should I use TOP10 cells or the LMG194 E. coli strain you offer for expression with my pBAD system?

Top10

Advantages:
- Saves time, can go directly from cloning to expression.
- The glycerol stock is more stable because these strains are endA- and recA-.

Disdvantages:
- This strain is not protease-deficient. Therefore, the protein may be degraded.

LMG194

Advantages:
- Grows well in minimal media, except M9.
-Have to transform the plasmid into the cells just for expression.
-RM medium with glucose to ensure low basal level of protein.

Disadvantages:
- Not protease-deficient. Therefore, the protein may be degraded.
- The glycerol stock may not be stable because this cell strain is not recA- or endA-.

What competent cells do you recommend I use for expression with my pBAD expression system?

We recommend using a competent cell strain that is araBADC- and araEFGH+, allowing transportation of L-arabinose, but not metabolizing it. This is important for expression studies, as the level of L-arabinose will be constant inside the cell and will not decrease over time. We offer our TOP10 competent cells, or our LMG194 E. coli strain.

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Citations & References (13)

Citations & References
Abstract
LuxS is required for persistent pneumococcal carriage and expression of virulence and biosynthesis genes.
Authors:Joyce EA, Kawale A, Censini S, Kim CC, Covacci A, Falkow S,
Journal:Infect Immun
PubMed ID:15102809
'Streptococcus pneumoniae causes several diseases, including otitis media, pneumonia, and meningitis. Although little is known about the regulation of or how individual pneumococcal factors contribute to these disease states, there is evidence suggesting that some factors are regulated by a cell-density-dependent mechanism (quorum sensing). Quorum sensing allows bacteria to couple ... More
Identification of a novel maturation mechanism and restricted substrate specificity for the SspB cysteine protease of Staphylococcus aureus.
Authors:Massimi I, Park E, Rice K, Muller-Esterl W, Sauder D, McGavin MJ,
Journal:J Biol Chem
PubMed ID:12207024
'The SspB cysteine protease of Staphylococcus aureus is expressed in an operon, flanked by the sspA serine protease, and sspC, encoding a 12.9-kDa protein of unknown function. SspB was expressed as a 40-kDa prepropeptide pSspB, which did not undergo autocatalytic maturation. Activity of pSspB was reduced compared with 22-kDa mature ... More
A dual-specificity aminoacyl-tRNA synthetase in the deep-rooted eukaryote giardia lamblia.
Authors:Bunjun S, Stathopoulos C, Graham D, Min B, Kitabatake M, Wang AL, Wang CC, Vivares CP, Weiss LM, Soll D
Journal:Proc Natl Acad Sci U S A
PubMed ID:11078517
'Cysteinyl-tRNA (Cys-tRNA) is essential for protein synthesis. In most organisms the enzyme responsible for the formation of Cys-tRNA is cysteinyl-tRNA synthetase (CysRS). The only known exceptions are the euryarchaea Methanococcus jannaschii and Methanobacterium thermoautotrophicum, which do not encode a CysRS. Deviating from the accepted concept of one aminoacyl-tRNA synthetase per ... More
RNA stem-loop enhanced expression of previously non-expressible genes.
Authors:Paulus M, Haslbeck M, Watzele M,
Journal:Nucleic Acids Res
PubMed ID:15163763
The key step in bacterial translation is formation of the pre-initiation complex. This requires initial contacts between mRNA, fMet-tRNA and the 30S subunit of the ribosome, steps that limit the initiation of translation. Here we report a method for improving translational initiation, which allows expression of several previously non-expressible genes. ... More
Shielding of the A1 Domain by the D'D3 Domains of von Willebrand Factor Modulates Its Interaction with Platelet Glycoprotein Ib-IX-V.
Authors:Ulrichts H, Udvardy M, Lenting PJ, Pareyn I, Vandeputte N, Vanhoorelbeke K, Deckmyn H,
Journal:J Biol Chem
PubMed ID:16373331
Soluble von Willebrand factor (VWF) has a low affinity for platelet glycoprotein (GP) Ibalpha and needs immobilization and/or high shear stress to enable binding of its A1 domain to the receptor. The previously described anti-VWF monoclonal antibody 1C1E7 enhances VWF/GPIbalpha binding and recognizes an epitope in the amino acids 764-1035 ... More
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