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pNiFty3-N-Lucia

pNiFty3-N-Lucia Unit size Cat. code Docs Qty Price
pNiFty3 - mIFN-β promoter - NF-κB - ZeocinR - Lucia
20 µg
pnf3-lc2
+-
$480.00

pNiFty3-N-Lucia plasmid is composed of three key elements: the mouse interferon beta minimal promoter, NF-κB and a secreted luciferase (Lucia) reporter gene.

pNiFty3-N-Lucia plasmid is selectable with Zeocin™ in both E. coli and mammalian cells, and can be used to generate stable clones.

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Specifications

Transcription factor binding sites: NF-κB (5x)
Minimal Promoter: mouse IFNβ promoter
Selection: Zeocin™
Reporter Gene: Lucia luciferase

These products are covered by a Limited Use License (See Terms and Conditions).

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Contents

pNiFty3-N-Lucia 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 and should be stored at -20°C.

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Description

Minimal promoter

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 [1].

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  [2].    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.

Reporter Gene

Lucia luciferase reporter gene: Lucia luciferase is a secreted luciferase with strong bioluminescent activity
The   Lucia reporter gene is ideal for promoter activity and gene   expression  studies. Lucia is encoded by a synthetic gene derived by   genetic  engineering of copepod luciferase genes. Lucia luciferase   activity can  be detected and quantified directly in the culture medium   of  transfected cells using InvivoGen's QUANTI-Luc™ detection reagent.

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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Details

pNiFty3-Lucia map

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