Patent ID: 6391641
Filing Date: 2002-05-21
Classification: C12N

Abstract:
A method of selecting for a missense mutant product comprising:(a) making a synthetic nucleic acid construct comprising an upstream coding sequence gene and a downstream reporter gene that are translationally coupled; (b) introducing mutations into the upstream gene, wherein step (a) and (b) may be performed in any order; (c) inserting the construct into a host cell; (d) selecting for a phenotype of the upstream coding sequence, which is different from the phenotype of the original upstream, and is caused by the mutation in the upstream coding sequence gene; and (e) selecting for the expression of the downstream coding sequence, wherein (i) a combination of the presence of the phenotype of the mutated upstream and expression of the downstream coding sequence gene product indicates that the mutation in the upstream coding sequence is a missense mutation, and wherein (ii) a combination of the presence of the phenotype of the mutated upstream and no expression of the downstream coding sequence gene product indicates that the mutation in the upstream coding sequence is a nonsense mutation.