Patent ID: 6479289
Filing Date: 2002-11-12
Classification: C12N,C12Q

Abstract:
A method of identifying an agent that disrupts interaction between a bait and a prey, comprising:(a) providing a mammalian cell comprising: (i) a reporter gene encoding a fluorescent polypeptide operably linked to a transcriptional regulatory sequence comprising a DNA binding site for a DNA-binding domain, wherein the reporter gene is stably integrated into the genome of the cell; (ii) a first nucleotide sequence encoding a bait fusion protein that comprises a DNA-binding domain and the bait; (iii) a second nucleotide sequence encoding a prey fusion protein comprising a transcriptional activation moiety and the prey; (b) contacting the mammalian cell with a test agent; (c) incubating the cell for 96 hours or less; and (d) detecting a decrease in expression of the reporter gene compared to the level of expression of the reporter gene in a mammalian control cell, thereby detecting an agent that disrupts interaction between the bait and the prey.