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pNiFty2 : mIFNα - ZeoR - SEAP
Inducible SEAP reporter plasmid- mIFNα promoter - ZeocinR
pNiFty2-IFA plasmid is composed of two key elements: the mouse interferon alpha 4 minimal promoter and a SEAP (Secreted alkaline phosphatase) reporter gene.
pNiFty2-IFA plasmids are selectable with Zeocin™ in both E. coli and mammalian cells, and can be used to generate stable clones.Back to the top
Minimal Promoter: mouse IFNα promoter
Reporter Gene: SEAPBack to the top
- pNiFty2-IFA plasmid is provided as 20 µg lyophilized DNA with 4 pouches of Fast-Media® (2 TB and 2 Agar), containing Zeocin™.
Products are shipped at room temperature
Stored at -20°C.Back to the top
The proximal promoters are shorter than 500 bp and contain transcription factor binding sites. Upon stimulation in 293 cells, their expression level remains undetectable. With the addition of repeated TFBS, the proximal promoters become inducible by the appropriate stimulus and drive the expression of the reporter gene.
mIFNα promoter is the mouse interferon alpha 4 minimal promoter . Transcription of mIFNα4 is mediated by a virus responsive element (VRE-A4) located in the promoter. VRE-A4 contains four cooperating DNA modules that
bind to IRF3 adn IRF7 . Co-expression of IFNα-SEAP with constitutively activated IRF3 (saIRF3) or IRF7 (saIRF7) in HEK293 cells led to a strong increase in SEAP expression.
SEAP reporter gene: Secreted alkaline phosphatase (SEAP) is a reporter widely used to study promoter activity or gene expression. SEAP expression can be rapidly and readily measured in supernatants of transfected cells. SEAP levels can be evaluated qualitatively with the naked eye and quantitatively using SEAP detection media, such as HEK-Blue™ Detection system or the SEAP Reporter Assay.
1. Braganca J. et al., 1997. Synergism between multiple virus-induced factor-binding elements involved in the differential expression of interferon A genes. J Biol Chem. 272(35):22154-62.
2. Morin P. et al., 2002. Preferential binding sites for interferon regulatory factors 3 and 7 involved in interferon-A gene transcription. J Mol Biol. 316(5):1009-22.