Reed, A.L., Yamazaki, H., Kaufman, J.D., Rubinstein, Y., Murphy, B., Johnson, A.C.
Notes: The cloned 752 amino acid GC-binding factor 2 (GCF2) was expressed in vitro using the TNT® Coupled Reticulocyte Lysate System. The synthesized product migrated at ~160kDa by SDS-PAGE when the predicted molecular weight was 83kDa. Expression of the protein from E.coli also produced a protein that migrated at ~160kDa. Mass spectrum analysis confirm a mass of 83kDa. The E.coli produced protein was further characterized by gel shift analysis with the AP-2 Consensus Oligonucleotide. Recombinant GCF2, AP-2 and SP1 transcription factors were analyzed for binding to the EGF receptor promoter by footprint analysis. The GCF2 protein was expressed with a EGF Receptor promoter-CAT reporter vector in CV-1 cells and found to increase CAT activity. (0510)