Detection of heterozygous truncating mutations in the BRCA1 and APC genes by using a rapid screening assay in yeast.

ABSTRACT: The detection of inactivating mutations in tumor suppressor genes is critical to their characterization, as well as to the development of diagnostic testing. Most approaches for mutational screening of germ-line specimens are complicated by the fact that mutations are heterozygous and that missense mutations are difficult to interpret in the absence of information about protein function. We describe a novel method using Saccharomyces cerevisiae for detecting protein-truncating mutations in any gene of interest. The PCR-amplified coding sequence is inserted by homologous recombination into a yeast URA3 fusion protein, and transformants are assayed for growth in the absence of uracil. The high efficiency of homologous recombination in yeast ensures that both alleles are represented among tra

SUBMITTER: Ishioka C 

PROVIDER: S-EPMC20108 | biostudies-literature | 1997 Mar

REPOSITORIES: biostudies-literature