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Citations & References (2)
Invitrogen™
T-REx™ Core Kit, with pcDNA™4/TO/myc-His© Vector
A Tetracycline-Regulated Expression System without Viral TransactivatorsThe T-REx™ System yields higher levels of induced expression than any other regulated mammalianRead more
| Catalog Number | Quantity |
|---|---|
| K103002 also known as K1030-02 | 1 kit |
Catalog number K103002
also known as K1030-02
Price (EUR)
2.820,00
Each
Quantity:
1 kit
Price (EUR)
2.820,00
Each
A Tetracycline-Regulated Expression System without Viral Transactivators
The T-REx™ System yields higher levels of induced expression than any other regulated mammalian expression system. It utilizes the complete CMV promoter and adds control elements from the bacterial tetracycline resistance operon to effectively repress and derepress transcription from one of the strongest mammalian promoter sequences known (1,2).
Specific Activation
The T-REx™ System uses a repressor mechanism that blocks transcription from the powerful CMV promoter in the absence of tetracycline. Because the T-REx™ System elements do not use viral transactivators, you can achieve high-level expression from the complete CMV promoter without secondary, non-specific activation of host genes.
The T-REx™ Mechanism
The T-REx™ transcriptional control elements are illustrated in Figure 1. Two tetracycline operator sequences (TetO2) have been inserted between the TATA box of the CMV promoter and the transcriptional start site. The TetO2 sequence itself has no effect on expression. When the tetracycline repressor protein (TR) is present, it effectively binds the TetO2 sites and blocks transcription initiation. Tetracycline added to the culture medium binds to, and changes the conformation of, the TR protein. This change causes the TR protein to release the TetO2 sites, derepressing transcription from the CMV promoter. The result is high-level expression of the gene of interest (Figure 2). Expression levels can be modulated based on the tetracycline concentration and can be induced to levels that are achieved with constitutive CMV expression vectors.
T-REx™ is a powerful inducible mammalian expression system that allows you to regulate expression from the complete human cytomegalovirus (CMV) enhancer-promoter. T-REx™inducible expression vectors offer the following features:
• Complete CMV enhancer-promoter sequence containing two copies of the tetracycline operator TetO2 sequence for high-level regulated expression
• Zeocin™ or hygromycin resistance gene for effective selection of stable mammalian cell lines
• Large multiple cloning site to simplify cloning
In addition, pcDNA™4/TO/myc-His offers a c-myc epitope for rapid detection of the recombinant protein with an Anti-myc Antibody and a polyhistidine (6xHis) sequence for simple purification of the recombinant protein with nickel-chelating resin and detection with Anti- His(C-term) Antibody.
The regulatory vector, pcDNA™6/TR, is provided for high-level expression of the tetracycline repressor (TR) protein. This vector expresses the Blasticidin resistance gene for rapid selection of mammalian cell lines that stably express the TR protein.
The T-REx™ System yields higher levels of induced expression than any other regulated mammalian expression system. It utilizes the complete CMV promoter and adds control elements from the bacterial tetracycline resistance operon to effectively repress and derepress transcription from one of the strongest mammalian promoter sequences known (1,2).
Specific Activation
The T-REx™ System uses a repressor mechanism that blocks transcription from the powerful CMV promoter in the absence of tetracycline. Because the T-REx™ System elements do not use viral transactivators, you can achieve high-level expression from the complete CMV promoter without secondary, non-specific activation of host genes.
The T-REx™ Mechanism
The T-REx™ transcriptional control elements are illustrated in Figure 1. Two tetracycline operator sequences (TetO2) have been inserted between the TATA box of the CMV promoter and the transcriptional start site. The TetO2 sequence itself has no effect on expression. When the tetracycline repressor protein (TR) is present, it effectively binds the TetO2 sites and blocks transcription initiation. Tetracycline added to the culture medium binds to, and changes the conformation of, the TR protein. This change causes the TR protein to release the TetO2 sites, derepressing transcription from the CMV promoter. The result is high-level expression of the gene of interest (Figure 2). Expression levels can be modulated based on the tetracycline concentration and can be induced to levels that are achieved with constitutive CMV expression vectors.
T-REx™ is a powerful inducible mammalian expression system that allows you to regulate expression from the complete human cytomegalovirus (CMV) enhancer-promoter. T-REx™inducible expression vectors offer the following features:
• Complete CMV enhancer-promoter sequence containing two copies of the tetracycline operator TetO2 sequence for high-level regulated expression
• Zeocin™ or hygromycin resistance gene for effective selection of stable mammalian cell lines
• Large multiple cloning site to simplify cloning
In addition, pcDNA™4/TO/myc-His offers a c-myc epitope for rapid detection of the recombinant protein with an Anti-myc Antibody and a polyhistidine (6xHis) sequence for simple purification of the recombinant protein with nickel-chelating resin and detection with Anti- His(C-term) Antibody.
The regulatory vector, pcDNA™6/TR, is provided for high-level expression of the tetracycline repressor (TR) protein. This vector expresses the Blasticidin resistance gene for rapid selection of mammalian cell lines that stably express the TR protein.
For Research Use Only. Not for use in diagnostic procedures.
Specifications
Constitutive or Inducible SystemInducible
Delivery TypeTransfection
For Use With (Application)Regulated Expression
Inducing AgentTetracycline
Product TypeT-Rex Core Cloning Kit with Vector
Quantity1 kit
Selection Agent (Eukaryotic)Zeocin™
VectorpcDNA
Cloning MethodRestriction Enzyme/MCS
Product LineT-REx, pcDNA
PromoterCMV/TO
Protein TagHis Tag (6x), c-Myc Epitope Tag
Unit SizeEach
Contents & Storage
The T-REx™ Core Kit includes 20 μg each of pcDNA™4/TO/myc-His A, B, and C; a positive control vector; pcDNA™6/TR; and forward and reverse sequencing primers.
All vectors are supercoiled and provided lyophilized.
Store all components at -20°C. All components are guaranteed stable for 6 months when properly stored.
All vectors are supercoiled and provided lyophilized.
Store all components at -20°C. All components are guaranteed stable for 6 months when properly stored.
Frequently asked questions (FAQs)
Why is sequential transfection recommended over co-transfection in the T-REx and GeneSwitch systems?
What is the main advantage of the GeneSwitch system over the T-REx system? And what is its main disadvantage?
What is the advantage of the Flp-In T-REx system over the T-REx system?
Can I use doxycycline instead of tetracycline as an inducer in the T-REx system?
I am planning to generate a T-REx cell line using pcDNA6/TR. Can I perform a western blot using antibodies to TetR to assess whether the cell line is expressing enough of TetR? Do you offer an antibody to TetR?
Citations & References (2)
Citations & References
Abstract
Interaction between Erbin and a Catenin-related protein in epithelial cells.
Journal:J Biol Chem
PubMed ID:11711544
'Integrity of epithelial tissues relies on the proper apical-basolateral polarity of epithelial cells. Members of the LAP (LRR and PDZ) protein family such as LET-413 and Scribble are involved in maintaining epithelial cell polarity in Caenorhabditis elegans and Drosophila melanogaster, respectively. We previously described Erbin as a mammalian LET-413 homologue
Human TRPC5 channel activated by a multiplicity of signals in a single cell.
Journal:J Physiol
PubMed ID:15254149
Here we explore the activation mechanisms of human TRPC5, a putative cationic channel that was cloned from a region of the X chromosome associated with mental retardation. No basal activity was evident but activity was induced by carbachol stimulation of muscarinic receptors independently of Ca2+ release. This is 'receptor activation',