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pNiFty2 : mIFNβ - ZeoR - SEAP
pNiFty2-IFB plasmid is composed of two key elements: the mouse interferon beta minimal promoter and a SEAP (Secreted alkaline phosphatase) reporter gene.
pNiFty2-IFB plasmid is selectable with Zeocin™ in both E. coli and mammalian cells, and can be used to generate stable clones.
Minimal Promoter: mouse IFNβ promoter
Reporter Gene: SEAP
pNiFty2-IFB 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.
mIFNβ promoter: The mouse interferon beta minimal promoter comprises several positive regulatory domains (PRDs) that bind different cooperating transcription factors such as NF-κB, IRF3 and IRF7 . Co-expression of IFNβ-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. Vodjdani G. et al. 1998. Structure and characterization of a murine chromosomal fragment containing the interferon beta gene. J Mol Biol. 204(2):221-31.Back to the top