pTrcHis A, B, & C Bacterial Expression Vectors
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Citations & References (54)
Invitrogen™

pTrcHis A, B, & C Bacterial Expression Vectors

Our pTrcHis A, B, & C vectors are designed to offer enhanced translation initiation and high-level expression in E. coli.Read more
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Catalog NumberQuantity
V3602020 μg
Catalog number V36020
Price (MXN)
-
Quantity:
20 μg
Our pTrcHis A, B, & C vectors are designed to offer enhanced translation initiation and high-level expression in E. coli. These vectors feature:

• High-level regulated transcription from the trc promoter
• Enhanced translation efficiency of eukaryotic genes in E. coli
• The lacO operator and lacIq repressor gene for transcriptional regulation in any E. coli strain

This particular vector offers:

• N-terminal polyhistidine (6xHis) tag for rapid purification with nickel-chelating resin and detection with an anti-HisG antibody
• N-terminal Xpress™ epitope for easy detection with an anti-Xpress™ antibody
• Enterokinase cleavage site for removal of fusion tag

For C-terminal polyhistidine tag and c-myc epitope, please see our pTrcHis2 A, B, & C Vector.
For Research Use Only. Not for use in diagnostic procedures.
Specifications
Antibiotic Resistance BacterialAmpicillin (AmpR)
CleavageEK (Enterokinase) Recognition Site
Constitutive or Inducible SystemInducible
Inducing AgentIPTG
Product TypeBacterial Expression Vector
Quantity20 μg
Selection Agent (Eukaryotic)None
VectorpTrc
Cloning MethodRestriction Enzyme/MCS
PromoterTrc, lacO
Protein TagThioredoxin
Unit Size20 µg
Contents & Storage
All vectors are provided in suspension. A TOP10 E. coli stab and positive expression control is also provided. Store vectors at -20°C. Store stab at room temperature. The vector is guaranteed stable for 6 months when properly stored.

Frequently asked questions (FAQs)

What are the advantages and/or disadvantages of using TOP10, DH5α, or other cloning strains versus BL21 Star (DE3) or BL21 (DE3) cells for expression with the pTrc system?

Top10, DH5α, other cloning strains

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

Disdvantages:
- If GOI is toxic, the cloning step can be difficult.
- These cloning strains are not protease-deficient; therefore, the protein may be degraded.

BL21 Star(DE3) or BL21 (DE3)

Advantages:
- These expression strains are protease-deficient.
- You have to transform the plasmid into an expression strain.
- The glycerol stock may be unstable because these expression strains are not endA- and recA-.
- The (DE3) part is wasted because the promoter does not need T7 RNA polymerase.

Find additional tips, troubleshooting help, and resources within our Protein Expression Support Center.

What competent cell strain can I use for expression with my pTrc Expression System?

The pTrc promoter is recognized by E. coli RNA polymerase, not T7 polymerase, and therefore can be expressed in any E. coli strain, not just BL21 strains. Therefore, you can use Top10, DH5α, etc. for expression. However, if your gene is toxic, the cloning step can be difficult as there is leakiness in expression.

Find additional tips, troubleshooting help, and resources within our Protein Expression Support Center.

How does glucose repress the pTrc promoter?

A transcriptional activator protein called CAP (catabolite activator protein) normally binds upstream of the trc promoter and activates transcription. This protein requires cAMP to bind the DNA. Adding glucose to the medium can reduce intracellular cAMP levels. Supplementing LB medium and agar plates with glucose will repress basal level transcription from the trc promoter. We recommend that you include 25 mM, 0.5% glucose in the selection medium to ensure stability of your insert.

Find additional tips, troubleshooting help, and resources within our Protein Expression Support Center.

What are the benefits of the pTrc expression system?

The pTrc promoter in the pTrc system is a strong hybrid promoter composed of the -35 region of the trp promoter and the -10 region of the lacUV5 promoter/operator. Expression of pTrc is repressed by the LacI protein and induced by IPTG.

Find additional tips, troubleshooting help, and resources within our Protein Expression Support Center.

Do I need to include a ribosomal binding site (RBS/Shine Dalgarno sequence) or Kozak sequence when I clone my gene of interest?

ATG is often sufficient for efficient translation initiation although it depends upon the gene of interest. The best advice is to keep the native start site found in the cDNA unless one knows that it is not functionally ideal. If concerned about expression, it is advisable to test two constructs, one with the native start site and the other with a Shine Dalgarno sequence/RBS or consensus Kozak sequence (ACCAUGG), as the case may be. In general, all expression vectors that have an N-terminal fusion will already have a RBS or initiation site for translation.

Find additional tips, troubleshooting help, and resources within our Protein Expression Support Center.

View all pTrcHis A, B, & C Bacterial Expression Vectors FAQs

Citations & References (54)

Citations & References
Abstract
Metaxin is a component of a preprotein import complex in the outer membrane of the mammalian mitochondrion.
Authors:Armstrong LC, Komiya T, Bergman BE, Mihara K, Bornstein P
Journal:J Biol Chem
PubMed ID:9045676
'Metaxin, a novel gene located between the glucocerebrosidase and thrombospondin 3 genes in the mouse, is essential for survival of the postimplantation mouse embryo. In this study, the subcellular location, domain structure, and biochemical function of metaxin were investigated. Anti-recombinant metaxin antibodies recognized 35- and 70- kDa proteins in mitochondria ... More
Dual DNA binding specificity of ADD1/SREBP1 controlled by a single amino acid in the basic helix-loop-helix domain.
Authors:Kim JB, Spotts GD, Halvorsen YD, Shih HM, Ellenberger T, Towle HC, Spiegelman BM
Journal:Mol Cell Biol
PubMed ID:7739539
'Adipocyte determination- and differentiation-dependent factor 1 (ADD1), a member of the basic helix-loop-helix (bHLH) family of transcription factors, has been associated with both adipocyte differentiation and cholesterol homeostasis (in which case it has been termed SREBP1). Using PCR-amplified binding analysis, we demonstrate that ADD1/SREBP1 has dual DNA sequence specificity, binding ... More
Characterization of protein kinase A and protein kinase C phosphorylation of the N-methyl-D-aspartate receptor NR1 subunit using phosphorylation site-specific antibodies.
Authors:Tingley WG, Ehlers MD, Kameyama K, Doherty C, Ptak JB, Riley CT, Huganir RL
Journal:J Biol Chem
PubMed ID:9030583
'Modulation of N-methyl-D-aspartate receptors in the brain by protein phosphorylation may play a central role in the regulation of synaptic plasticity. To examine the phosphorylation of the NR1 subunit of N- methyl-D-aspartate receptors in situ, we have generated several polyclonal antibodies that recognize the NR1 subunit only when specific serine ... More
Ca2+/calmodulin binds to and modulates P/Q-type calcium channels [see comments]
Authors:Lee A, Wong ST, Gallagher D, Li B, Storm DR, Scheuer T, Catterall WA
Journal:Nature
PubMed ID:10335845
'Neurotransmitter release at many central synapses is initiated by an influx of calcium ions through P/Q-type calcium channels, which are densely localized in nerve terminals. Because neurotransmitter release is proportional to the fourth power of calcium concentration, regulation of its entry can profoundly influence neurotransmission. N- and P/Q-type calcium channels ... More
Oligomerization-dependent association of the SAM domains from Schizosaccharomyces pombe Byr2 and Ste4.
Authors: Ramachander Ranjini; Kim Chongwoo A; Phillips Martin L; Mackereth Cameron D; Thanos Christopher D; McIntosh Lawrence P; Bowie James U;
Journal:J Biol Chem
PubMed ID:12171939
'SAM (sterile alpha motif) domains are protein-protein interaction modules found in a large number of regulatory proteins. Byr2 and Ste4 are two SAM domain-containing proteins in the mating pheromone response pathway of the fission yeast, Schizosaccharomyces pombe. Byr2 is a mitogen-activated protein kinase kinase kinase that is regulated by Ste4. ... More
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