Invitrogen™ GeneSwitch™ Mammalian Expression Kit, complete kit

Description

Includes

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

Offers exceptionally low uninduced and high induced expression levels in mammalian cells

• Expression vector pGene/V5-His provides minimal promoter, GAL4-E1b, consisting of binding sites for yeast Gal4 DNA binding protein followed by TATA sequence from Adenovirus E1b promoter
• Without additional factors, GAL4-E1b promoter is transcriptionally silent
• To activate transcription from GAL4-E1b promoter, GeneSwitch regulatory protein is expressed from minimal TK promoter on pSwitch vector
• GeneSwitch protein has three functional domains: Gal4 DNA Binding Domain (Gal4-DBD) to bind regulatory protein to 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 NFØB transcription factor p65 (p65AD) to activate transcription from silent GAL4-E1b promoter
• In absence of mifepristone, conformation of hPR-LBD region prevents GeneSwitch regulatory protein from activating transcription from GAL4-E1b promoter
• When mifepristone is added and binds hPR-LBD region, GeneSwitch regulatory protein assumes conformation that permits to stimulate transcription from GAL4-E1b promoter
• Induction of GeneSwitch System leads to activation of gene of interest on vector pGene/V5-His
• In addition, four Gal4 binding sites upstream of minimal HSV TK promoter on pSwitch can bind pSwitch regulatory protein
• Therefore, adding mifepristone up-regulates production of regulatory protein
• Increased levels of GeneSwitchregulatory protein result in induction of gene of interest from pGene/V5-His to levels that can approach those of viral promoters
• Provides expression vector pGene/V5-His with minimal, synthetic GAL4-E1b promoter that is transcriptionally silent until activated

• In addition, GeneSwitch protein upregulates its own expression, leading to amplified expression of gene of interest from pGene/V5-His
• 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
• Zeocin resistance gene for effective selection in both mammalian cells and E. coli

Mammalian Expression, Protein Expression, Proteins, Expression, Isolation and Analysis, Regulated Expression

Specifications

• Constitutive or Inducible System: Inducible
• Delivery Type: Transfection
• Inducing Agent: Mifepristone
• Promoter: E1b (GAL4-E1b)
• Product Type: Mammalian Expression Kit
• Selection Agent (Eukaryotic): Zeocin™
• Content And Storage: 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, and the pSwitch vector. The kit also includes forward and reverse sequencing primers and 100 μg of the inducing agent mifepristone. The GeneSwitch™ Complete Kit includes the Core Kit with 1 g hygromycin and 1 g Zeocin™ at considerable savings. Hygromycin should be stored at +4°C and Zeocin™ at -20°C. Zeocin™ should be protected from exposure to light. All reagents are stable for 6 months when properly stored.
• Protein Tag: His Tag (6x), V5 Epitope Tag
• Cloning Method: Restriction Enzyme/MCS
• Quantity: 1 kit
• Vector: pGene
• Product Line: GeneSwitch™
• For Use With (Application): Regulated Expression

For Research Use Only. Not for use in diagnostic procedures.