The recently-determined sequence of the genome of bakers' yeast (S. cerevisiae) revealed that almost two-thirds of the genes of this well-studied organism had escaped detection. Thus, we know almost nothing about the function of most of the genes that comprise this simple eukaryotic cell. To catalyze approaching the ultimate goal of a comprehensive understanding of the function of all the proteins that constitute a eukaryotic cell, we propose to construct quickly the complete set of mutants deleted for each of the approximately 6000 genes of this important model organism. It seems certain that wide distribution of this collection of mutants will greatly accelerate the pace of discovery of gene function. This goal will be achieved by a consortium of 5 laboratories, each disrupting 300 genes in each of two year, yielding 3000 mutants. The other 3000 genes of the organism will be disrupted by our European and Canadian colleagues. The mutants will be screened for a few standard phenotypes to provide necessary information for the users of the mutant collection, then distributed widely to all interested scientists for more detailed and complete analysis. We propose to replace the coding sequence of each gene with the KanMx gene encoding resistance to the drug gentamycin. In addition, a novel sequence tag will be introduced into each of the mutants to uniquely identify them. Non-essential genes will be disrupted in haploids of both mating types; essential and nearly essential genes will be disrupted in diploids. Gene disruption will be accomplished using a facile and popular PCR-based technique that will enable construction of the complete set of mutants in two years. In the final year of the project, we will enhance the value of the mutant collection by constructing conditionally-expressed versions of essential and nearly essential genes, constructing approximately 750 double mutants deleted for both members of a duplicated gene pair, and disrupting small genes as they are revealed.

Agency
National Institute of Health (NIH)
Institute
National Human Genome Research Institute (NHGRI)
Type
Research Project (R01)
Project #
5R01HG001627-02
Application #
2674260
Study Section
Special Emphasis Panel (ZHG1-HGR-N (J1))
Program Officer
Feingold, Elise A
Project Start
1997-04-01
Project End
2000-03-31
Budget Start
1998-04-01
Budget End
1999-03-31
Support Year
2
Fiscal Year
1998
Total Cost
Indirect Cost
Name
Washington University
Department
Genetics
Type
Schools of Medicine
DUNS #
062761671
City
Saint Louis
State
MO
Country
United States
Zip Code
63130