Patent ID: 6468746
Filing Date: 2002-10-22
Classification: C12N,C40B

Abstract:
A method for selecting genes which code for stabilized proteins, which comprises the following steps:a) modification of an infectious replicable gene package (IRG) so that its infection of a host cell can be suppressed, as described under (b) and (d); b) insertion of DNA sequences which code for a repertoire of mutants of a protein (PT) into the genome of the IRG so that said mutants of the protein (PT) are incorporated as fusion proteins in a protein necessary for infection of a host organism; c) expression of the IRGs with a repertoire of mutants of the protein (PT) in a recombinant host organism; d) subjection of the IRGs to partial proteolysis during or after incubation under solvent conditions which influence the conformational stability of the protein (PT) so that the IRGs which are not stable under said solvent conditions are cleaved during the partial proteolysis and therefore loose their infectivity, and so that the IRGs which are stable under said solvent conditions and therefore not cleaved during the partial proteolysis retain their infectivity, and wherein the partial proteolysis and incubation are done in the presence of a ligand, inhibitor or substrate analog for the protein (PT); e) infection of host organisms after treatment according to (d) and amplification of the host organisms; f) isolation of IRGs which contain genes which code for stable proteins (PT) from said host organisms; g) use of the recovered IRGs repeatedly in the complete process from (b) to (f); h) isolation of the genes from the recovered IRGs.