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Proc Natl Acad Sci U S A

Title:		Dominant genetics using a yeast genomic library under the control of a strong inducible promoter
Author(s):		Ramer SW; Elledge SJ; Davis RW;
Address:		"Department of Biochemistry, Stanford University School of Medicine, CA 94305"
Journal Title:		Proc Natl Acad Sci U S A
Year:		1992
Volume:		89
Issue:		23
Page Number:		11589 - 11593
DOI:		10.1073/pnas.89.23.11589
ISSN/ISBN:		0027-8424 (Print) 1091-6490 (Electronic) 0027-8424 (Linking)
Abstract:		"In Saccharomyces cerevisiae, numerous genes have been identified by selection from high-copy-number libraries based on 'multicopy suppression' or other phenotypic consequences of overexpression. Although fruitful, this approach suffers from two major drawbacks. First, high copy number alone may not permit high-level expression of tightly regulated genes. Conversely, other genes expressed in proportion to dosage cannot be identified if their products are toxic at elevated levels. This work reports construction of a genomic DNA expression library for S. cerevisiae that circumvents both limitations by fusing randomly sheared genomic DNA to the strong, inducible yeast GAL1 promoter, which can be regulated by carbon source. The library obtained contains 5 x 10(7) independent recombinants, representing a breakpoint at every base in the yeast genome. This library was used to examine aberrant gene expression in S. cerevisiae. A screen for dominant activators of yeast mating response identified eight genes that activate the pathway in the absence of exogenous mating pheromone, including one previously unidentified gene. One activator was a truncated STE11 gene lacking approximately 1000 base pairs of amino-terminal coding sequence. In two different clones, the same GAL1 promoter-proximal ATG is in-frame with the coding sequence of STE11, suggesting that internal initiation of translation there results in production of a biologically active, truncated STE11 protein. Thus this library allows isolation based on dominant phenotypes of genes that might have been difficult or impossible to isolate from high-copy-number libraries"
Keywords:		"Base Sequence DNA, Fungal/genetics Galactose *Gene Expression Regulation, Fungal *Genes, Dominant *Genes, Fungal Genetic Complementation Test Genetic Vectors Genomic Library Mating Factor Molecular Sequence Data Oligodeoxyribonucleotides/chemistry Peptide;"
Notes:		"MedlineRamer, S W Elledge, S J Davis, R W eng 2TRGM07599/RG/CSR NIH HHS/ GM28428/GM/NIGMS NIH HHS/ R37HG00198/HG/NHGRI NIH HHS/ Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. 1992/12/01 Proc Natl Acad Sci U S A. 1992 Dec 1; 89(23):11589-93. doi: 10.1073/pnas.89.23.11589"