Bai, G. and Kusiak, J.W.
Notes: Reporter vectors (produced with the pGL2 Basic Vector) were transiently transfected into PC12 cells and promoter activity was measured in response to 2.5S NGF. Promoter constructs were generated by PCR, subcloned into the pGEM®-T Vector and sequenced. The pGEM®-luc Vector was used to generated an antisense luc RNA probe used in RNase protection assays (using RNase ONE™ Ribonuclease) to follow the transcription of luciferase DNA following NGF treatment. The early growth reaction transcription factor and variants were expressed in the TNT® SP6 Coupled Reticulocyte Lysate System and used to perform gel shifts with promoter elements. The SP1 Consensus Oligonucleotide was used to compete transcription factor binding to the promoter elements. (1493)