Patent ID: 6972178
Filing Date: 2005-12-06
Classification: G01N

Abstract:
1. A two-hybrid screening method for identifying a pair of molecules that bind to a proline-rich nuclear receptor co-regulatory protein (PNRC) molecule via the binding sequence of SEQ ID NO:5, wherein said PNRC molecule is selected from the group consisting of SEQ ID NO:8 and SEQ ID NO:9, wherein said PNRC molecule comprises said binding sequence of SEQ ID NO:5, wherein said PNRC molecule binds a liganded nuclear receptor and wherein said pair of molecules consists of a nuclear hormone receptor peptide molecule and a ligand molecule, the method comprising: a. providing a cell culture (1) that expresses a bait PNRC fusion molecule which comprises said PNRC molecule and (2) that comprises a reporter gene, wherein the expression of said reporter gene depends on binding of said bait PNRC fusion molecule to said nuclear hormone receptor peptide molecule; b. transfecting said cell culture with an expression library of nucleic acids that encode said nuclear hormone receptor peptide molecules to be screened, to produce a library of cells that express said nuclear hormone receptor peptide molecules to be screened and said bait PNRC; c. dividing said cells into first and second portions; d. growing said first portion of said cells in the presence of said ligand molecule to be screened; e. growing said second portion of said cells in the absence of said ligand molecule to be screened; f. comparing the level of expression of said reporter gene in the cells of said first and second portions; and g. if cells of said first and second portions express said reporter gene at different levels, identifying said pair of molecules as binding to PNRC.