Novel Approach to the Development of Highly Specific Inhibitors of EFG, a Critical Transcription Factor in Prostate Cancer; Annual rept. 15 Feb 2007-14 Feb 2008

摘要

We have created a series of six constructs to express varying length fragments of ERG. Expression and purification protocols have been established for all six fragments. Using fluorescein-labeled oligonucleotides, we have developed a fluorescence polarization based assay for measuring the DNA binding of the ERG fragments to a functional DNA element. Using this assay, we have shown that longer fragments of ERG show decreased DNA binding, i.e. are auto- inhibited. Based on this data, we have focused efforts on producing one fragment in the labeled form necessary for NMR structural studies.