pIκB-EGFP

Description:

The pIκB-EGFP Vector encodes the IκB-EGFP Signaling Probe, which is a fusion of enhanced green

fluorescent protein （EGFP） and IκB. IκB is an inhibitor of NFκB, a transcription factor involved in the

immune response and inflammatory diseases. When the NFκB pathway is inactive, IκB and NFκB

exist as an inactive complex in the cytosol. Upon stimulation, IκB is degraded. In cells transfected

with pIκB-EGFP, degradation of the IκB-EGFP fusion protein is observed as a decrease in EGFP

fluorescence （1）. The IκB-EGFP Signaling Probe is constitutively expressed and resides in the

cytosol.

EGFP is a red-shifted, human codon-optimized variant of GFP （2–6） that has been engineered for

brighter fluorescence and higher expression in mammalian cells. Its excitation maximum is 488 nm

and emission maximum is 509 nm. For more information on the properties of EGFP, please refer to

the BD Living Colors™ User Manual （PT2040-1） included with the vector.

Expression of the IκB-EGFP is driven by the human CMV immediate-early promoter. The SV40 poly-A

sequence directs proper processing of the 3′ end of the fusion construct. The vector backbone also

contains an SV40 origin for replication in mammalian cells expressing the SV40 T-antigen. A

neomycin resistance cassette （Neor

） allows kanamycin selection in E. coli and neomycin （G418）

selection in eukaryotic cells. This cassette consists of the SV40 early promoter, the neomycin/

kanamycin resistance gene of Tn5, and poly-A signals from the Herpes simplex virus thymidine

kinase （HSV TK） gene. The vector backbone also provides a pUC origin of replication for

propagation in E. coli and an f1 origin for single-stranded DNA production.