I am Jason Li, a research scientist at Clontech. We have generate a set of cis-reporters with different reporters, including destabilized EGFP. EGFP is too stable and not good for induction study. The destabilized form of EGFP that we generated has a half-life of 2 hours or less. This destabilized EGFP gave nice time-course induction. Please see our recent publication on JBC(Dec,1998). We currently constructed a NFAT-Luc reporter. I would like to have yours for comparison if it is possible. Thanks.
Jason Li, Ph.D.
Clontech Laboratories, Inc.
Phone: 650-424-8222 X1134
>>> TJ Murphy <tmurphy at pharm.emory.edu> 02/04/99 02:09PM >>>
If you got an NFAT/Luc reporter plasmid from Stefan Ho, an NFAT
re/minimal IL-2 promoter fragment drops out as ~200 bp BamHI-HindIII
fragment. It would be trivial to replace that into any eGFP expression
vector you have. The HindIII should be next to the protein reading
Beware, eGFP is very stable! Fold induction could be minimal at best if
you've got some leak in the cells.
T.J. Murphy, Ph.D. 404-727-2467
Assistant Professor 404-727-0365 (fax)
Department of Pharmacology tmurphy at pharm.emory.edu
Emory University School of Medicine
5031 O.W. Rollins Research Building
Atlanta, GA 30322
"If we're not driving paradigms, then we're out driving Titleists"