Patent ID: 6174722
Filing Date: 2001-01-16
Classification: C12N,C12P,C12Q

Abstract:
A method of identifying a pharmacologically active substance, comprisinga) providing a DNA template which comprises a target DNA sequence under the control of at least one gene-regulatory element,b) mixing with the template a nuclear extract and at least one labeled nucleotide,c) mixing with the template a candidate pharmacologically active substance,d) optionally removing the proteins from the reaction mixture after in vitro transcribing the DNA template,e) binding the labeled transcript to a solid matrix,f) removing the excess labeled nucleotides, andg) determining the amount of labeled transcript, relative to a control samplewherein said DNA template comprises unique cleavage sites for the restriction endonucleases PstI, EcoRI, SacI, KpnI, SacII, BamHI, SwaI, part of the plasmid pUC19, five binding sites for the yeast Gal4 protein, the ""TATA"" box of the human T-cell receptor V.beta.8.1 between the SacII and BamHI restriction sites, the initiator region of the adenovirus major late promoter between the BAMHI and SwaI restriction sites, and a G-free sequence with a length of approximately 800 base pairs.