Invitrogen™ GeneSwitch™ Mammalian Expression Kit, complete kit

Invitrogen™ GeneSwitch™ Mammalian Expression Kit, complete kit

The GeneSwitch™ System for inducible mammalian expression is ideal for experiments that require the absolute lowest uninduced expression levels. The expression vector pGene/V5-His provides a minimal promoter, GAL4-E1b, consisting of the binding sites for the yeast Gal4 DNA binding protein followed by the TATA sequence from the Adenovirus E1b promoter. Without additional factors, the GAL4-E1b promoter is transcriptionally silent.

To activate transcription from the GAL4-E1b promoter, the GeneSwitch™ regulatory protein is expressed from a minimal TK promoter on the pSwitch vector. 

The GeneSwitch™ protein has three functional domains:

• Gal4 DNA Binding Domain (Gal4-DBD) to bind the regulatory protein to the GAL4-E1b promoter
• Truncated human Progesterone Receptor Ligand Binding Domain (hPR-LBD) that undergoes a conformational change when it binds the progesterone antagonist, mifepristone
• Transcription activation domain from the NFØB transcription factor p65 (p65AD) to activate transcription from the silent GAL4-E1b promoter

The GeneSwitch™ Kit offers exceptionally low uninduced and high induced expression levels in mammalian cells. The system provides the expression vector pGene/V5-His with the minimal, synthetic GAL4-E1b promoter that is transcriptionally silent until activated. The GeneSwitch™ regulatory protein binds the GAL4-E1b promoter to activate transcription upon the addition of mifepristone. In addition, the GeneSwitch™ protein upregulates its own expression, leading to amplified expression of the gene of interest from pGene/V5-His. 

Features

C-terminal V5 epitope tag for detection with Anti-V5 Antibodies

C-terminal polyhistidine (6xHis) tag for rapid purification with nickel-chelating resin and detection with Anti-His(C-term) Antibodies

The Zeocin™ resistance gene for effective selection in both mammalian cells and E. coli

Contents

• The GeneSwitch™ System Core Kit includes 20 µg of each of the following vectors: pGene/V5-His (A, B, and C)
• the expression control vector pGene/V5-His/lacZ
• the pSwitch vector
• forward and reverse sequencing primers and 100 µg of the inducing agent mifepristone
• the Core Kit with 1 g hygromycin and 1 g Zeocin™ at considerable savings