BRAND / VENDOR: Thermo Fisher

Thermo Fisher, V102020, pcDNA™4/TO Mammalian Expression Vector

CATALOG NUMBER: V102020

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Product Description

pcDNA™4/TO Mammalian Expression Vector A Tetracycline-Regulated Expression System without Viral TransactivatorsThe 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 ActivationThe 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™ MechanismThe 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 TetO2sequence itself has no effect on expression. When the tetracycline repressor protein (TR) is present, it effectively binds the TetO2sites 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 TetO2sites, 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 TetO2sequence for high-level regulated expression
• Zeocin™ or hygromycin resistance gene for effective selection of stable mammalian cell lines
• Large multiple cloning site to simplify cloningIn addition, pcDNA™4/TO/myc-His offers ac-mycepitope for rapid detection of the recombinant protein with an Anti-mycAntibody 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.
Constitutive or Inducible System: Inducible
Delivery Type: Transfection
For Use With (Application): Regulated Expression
Inducing Agent: Tetracycline
Product Type: Mammalian Expression Vector
Quantity: 20 μg
Selection Agent (Eukaryotic): Zeocin™
Vector: pcDNA
Cloning Method: Restriction Enzyme/MCS
Product Line: T-REx™, pcDNA™
Promoter: CMV/TO
Protein Tag: Untagged
Unit Size: 20 µg

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