Patent ID: 7297491
Filing Date: 2007-11-20
Classification: C12N,C12Q,G01N

Abstract:
1. A method for detecting an interaction between a test polypeptide and a DNA sequence, comprising: i providing a population of host cells wherein each cell contains (a) a first reporter gene operably linked to a transcriptional regulatory sequence which includes a binding site for a weak zinc-finger DNA-binding domain, (b) a second reporter gene operably linked to a transcriptional regulatory sequence which includes a binding site for the weak zinc-finger DNA-binding domain, (c) a chimeric gene which encodes a fusion protein, the fusion protein including a test polypeptide, the weak zinc finger DNA-binding domain and an activation tag, wherein binding of the weak zinc finger DNA-binding domain of (c) to the binding sites of (a) or (b) does not cause a significant increase in the expression of the first reporter gene or the second reporter gene; wherein expression of the first reporter gene results in a first detectable signal; wherein expression of the second reporter gene results in a second detectable signal; wherein a non-specific interaction between a test polypeptide of the fusion protein and the transcriptional regulatory sequence of the first and second reporter genes results in an increased level of expression of the first and second reporter genes; wherein a specific interaction between a test polypeptide of the fusion protein and the transcriptional regulatory sequence of the first or second reporter gene results in a desired level of expression of either the first or second reporter gene; and ii isolating host cells comprising a fusion protein that specifically interacts with the transcriptional regulatory sequence the first or second reporter gene exhibiting a desired level of expression of the first or second reporter gene, thereby detecting an interaction between the test polypeptide and the transcriptional regulatory sequence.