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pNiFty3-I-Lucia

pNiFty3-I-Lucia Unit size Cat. code Docs Qty Price
pNiFty3 - mIFN-β promoter - ISRE - ZeocinR - Lucia
20 µg
pnf3-lc4
+-
$480.00

pNiFty3-I-Lucia plasmid is composed of three key elements:  the mouse interferon beta minimal promoter, five ISRE transcription factor binding sites and a secreted luciferase (Lucia) reporter gene.

pNiFty3-I-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: ISRE (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-I-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)

ISRE  binding site: PRRs involved in the antiviral response induce the  activation of interferon regulatory factors (IRFs) and the production of  type I interferons (IFNs). IFNs trigger the formation of the ISGF3  complex which contains signal transducer and activator of transcription  (STAT) 1, STAT2 and IRF9. ISGF3 and IRFs bind to specific nucleotide  sequences called interferon-stimulated response elements (ISREs;  AGTTTCNNTTTCC) in the promoter of IFN-stimulated genes (ISGs) leading to  their activation [2].

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. Wesoly J. et al., 2007. STAT activation and  differential complex formation dictate selectivity of interferon  responses. Acta Biochim Pol. 54(1):27-38.

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Details

pNiFty3-Lucia map

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