A Novel Yeast Genomics Method for Identifying New Breast Cancer Susceptibility Genes
Abstract
We are attempting to identify novel genes in the yeast S. cerevisiae that confer gross chromosomal instability (GCI) when deleted. Using a collection of yeast strains carrying deletion of all non-essential open reading frames, we will transfect a yeast artificial chromosome (YAC) as a reporter for GCI frequency and determine the quantitative impact of the loss of each gene function. We have constructed the redesigned reporter with all of the components for selection and maintenance. The URA3 gene results in sensitivity to 5-FOA and the methionine regulated BAX gene induces apoptosis in yeast in individual and pooled deletion strains. We have modified the original YAC to incorporate the NatMX (nourseothricin resistance) marker, which has been shown to overcome the defects in the original design. We have transfected the final YAC construct into yeast deletions strains with well documented genomic instabilities that span the current spectrum of measurable frequencies. We are transfecting the entire collection of deletion strains to assess the pool of yeast deletion strains to identify genes affecting GCI to determine the mammalian orthologs of these genes as well as those of interacting partners using in silico methods. There is a crucial need to find new candidate genes for breast cancer susceptibility in women and identifying these genes can further improve monitoring and treatment guidelines for women with these mutations.
Document Details
- Document Type
- Technical Report
- Publication Date
- May 01, 2006
- Accession Number
- ADA457723
Entities
People
- J. M. Brown
Organizations
- Stanford University