Patent ID: 6242183
Filing Date: 2001-06-05
Classification: C07K,C12N,C12Q

Abstract:
A method of determining whether a first candidate protein interacts with a second candidate protein, comprising measuring reporter gene expression in a Saccharomyces or mammalian host cell which contains(i) said reporter gene operably linked to a DNA-binding-protein recognition site,(ii) a first fusion gene which expresses a first fusion protein, said first fusion protein comprising said first candidate protein covalently bonded to a binding moiety which specifically binds to said DNA-binding-protein recognition site, and(iii) a second fusion gene which expresses a second fusion protein, said second fusion protein comprising said second candidate protein covalently bonded to a gene activating moiety,wherein either said first or said second candidate protein has reduced structural flexibility due to covalent bonding of the amino and carboxy termini of said candidate protein to a conformation-constraining protein which is heterologous to said candidate protein, andwhereby expression of said reporter gene is a measure of an interaction between said first and said second candidate proteins.