MUTATION DETECTION BY A 2-HYBRID ASSAY

Yeast-based assays have been developed to detect inactivating mutation s in human genes, but these assays generally rely on the human protein having a biological function in yeast. We describe a simple method to detect mutations by virtue of their ability to abolish a protein-prot ein interaction in the yeast two-hybrid assay. By the use of direct re combinational cloning in yeast of a reverse transcription-PCR product followed by a simple growth selection this method distinguished both h omozygous and heterozygous mutations in the p53 tumor suppressor gene. This approach should be applicable to many human genes whose encoded proteins have suitable partners in the two-hybrid assay.