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pNiFty3 - mIFN-β promoter - NF-κB - ZeocinR - SEAP
Inducible SEAP reporter plasmid - mIFN-β promoter - NF-κB - ZeocinR
pNiFty3-N-SEAP plasmid is composed of three key elements: the mouse interferon beta minimal promoter, NF-κB and a SEAP (Secreted alkaline phosphatase) reporter gene.
pNiFty3-N-SEAP plasmid is selectable with Zeocin™ in both E. coli and mammalian cells, and can be used to generate stable clones.Back to the top
Transcription factor binding sites: NF-κB (5x)
Minimal Promoter: mouse IFNβ promoter
Reporter Gene: SEAPBack to the top
- pNiFty3-N-SEAP 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.
Products should be 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.
IFN-β promoter: the mouse IFN-β minimal promoter comprises several positive regulatory domains that bind different cooperating transcription factors such as NF-kB, IRF3 and IRF7 .
Transcription factor binding sites (TFBS)
NF-kB binding site: Nuclear factor (NF)-κB is a “rapid-acting” primary transcription factor activated by a wide variety of PRRs. NF-κB is a protein complex that belongs to the Rel-homology domain-containing protein family. The prototypical NF-κB is composed of the p65(RelA) and p50 subunits . NF-κB binds specific decameric DNA sequences (GGGRNNYYCC, R-purine Y=pyrimidine) and activates genes involved in the regulation of the innate and adaptative immune response.
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. Vodjdani G. et al., 1988. Structure and characterization of a murine chromosomal fragment containing the interferon beta gene. J Mol Biol. 204(2):221-31.
2. Kawai T. & Akira S., 2007. Signaling to NF-kappaB by Toll-like receptors. Trends Mol Med. 13(11):460-9.
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