Patent ID: 6294330
Filing Date: 2001-09-25
Classification: C07K,C12N,G01N,Y02A

Abstract:
A method employing a Protein Complementation assay/Universal Reporter System (PCA/URS) for detecting and screening for ligands of a cellular receptor, which method comprises:a) generating a first nucleic acid vector encoding a first fusion product comprising:i) a first fragment of a first PCA/URS reporter molecule, andii) a second molecule, fused to said first fragment, which comprises a first subdomain of a cellular receptor molecule of interest;b) generating a second nucleic acid vector encoding a second fusion product comprising:i) a second fragment of said first PCA/URS reporter molecule, andii) a third molecule, fused to said second fragment, which comprises a second subdomain of said cellular receptor, and where said second subdomain may be the same as said first subdomain in the case of a homodimeric cellular receptor, or different from said first subdomain in the case of a heterodimeric cellular receptor; or a receptor coactivator or a protein;c) transfecting prokaryotic or eukaryotic cells with said first and second nucleic acid vectors; andd) testing said transfected cells for activity of said PCA/URS reporter molecule, said activity indicating reassociation of the first and second fragments of the PCA/URS reporter molecule mediated by the interaction of said first and second subdomains of the cellular receptor molecule; said association being induced by binding said receptor to said ligand.