Cloning and characterization of a gene which determines osmotic stability in Saccharomyces cerevisiae
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| Published in | Molecular and Cellular Biology Vol. 11; no. 8; pp. 4235 - 4243 |
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| Main Authors | , , , |
| Format | Journal Article |
| Language | English |
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Washington, DC
American Society for Microbiology
01.08.1991
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| Subjects | |
| Online Access | Get full text |
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| AbstractList | The srb1-1 mutation of Saccharomyces cerevisiae is an ochre allele which renders the yeast dependent on an osmotic stabilizer for growth and gives the cells the ability to lyse on transfer to hypotonic conditions. A DNA fragment which complements both of these phenotypic effects has been cloned. This clone contains a functional gene which is transcribed into a 2.3-kb polyadenylated mRNA molecule. Transformation of yeast strains carrying defined suppressible alleles demonstrated that the cloned fragment does not contain a nonsense suppressor. Integrative transformation and gene disruption experiments, when combined with classical genetic analysis, confirmed that the cloned fragment contained the wild-type SRB1 gene. The integrated marker was used to map SRB1 to chromosome XV by Southern hybridization and pulsed-field gel electrophoresis. A disruption mutant created by the insertion of a TRP1 marker into SRB1 displayed only the lysis ability phenotype and was not dependent on an osmotic stabilizer for growth. Lysis ability was acquired by growth in (or transfer to) an osmotically stabilized environment, but only under conditions which permitted budding. It is inferred that budding cells lyse with a higher probability and that weak points in the wall at the site of budding are involved in the process. The biotechnological potential of the cloned gene and the disruption mutant is discussed. Article Usage Stats Services MCB Citing Articles Google Scholar PubMed Related Content Social Bookmarking CiteULike Delicious Digg Facebook Google+ Mendeley Reddit StumbleUpon Twitter current issue Spotlights in the Current Issue MCB About MCB Subscribers Authors Reviewers Advertisers Inquiries from the Press Permissions & Commercial Reprints ASM Journals Public Access Policy MCB RSS Feeds 1752 N Street N.W. • Washington DC 20036 202.737.3600 • 202.942.9355 fax • journals@asmusa.org Print ISSN: 0270-7306 Online ISSN: 1098-5549 Copyright © 2014 by the American Society for Microbiology. For an alternate route to MCB .asm.org, visit: MCB The srb-1 mutation of Saccharomyces cerevisiae is an ochre allele which renders the yeast dependent on an osmotic stabilizer for growth and gives the cells the ability to lyse on transfer to hypotonic conditions. A DNA fragment which complements both of these phenotypic effects has been cloned. This clone contains a functional gene which is transcribed into a 2.3-kb polyadenylated mRNA molecule. Transformation of yeast strains carrying defined suppressible alleles demonstrated that the cloned fragment does not contain a nonsense suppressor. Integrative transformation and gene disruption experiments, when combined with classical genetic analysis, confirmed that the cloned fragment contained the wild-type SRB1 gene. The integrated marker was used to map SRB1 to chromosome XV by Southern hybridization and pulsed-field gel electrophoresis. A disruption mutant created by the insertion of a TRP1 marker into SRB1 displayed only the lysis ability phenotype and was not dependent on an osmotic stabilizer for growth. |
| Author | L I Stateva P V Venkov L J Trueman S G Oliver |
| AuthorAffiliation | Institute of Molecular Biology, Bulgarian Academy of Sciences, Sofia |
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| Keywords | Fungi Regulation(control) Yeast Gene Restriction map Ascomycetes Mutation Molecular cloning Osmotic pressure Saccharomyces cerevisiae Thallophyta |
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Mendeley... The srb1-1 mutation of Saccharomyces cerevisiae is an ochre allele which renders the yeast dependent on an osmotic stabilizer for growth and gives the cells... The srb-1 mutation of Saccharomyces cerevisiae is an ochre allele which renders the yeast dependent on an osmotic stabilizer for growth and gives the cells the... |
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| SubjectTerms | Biological and medical sciences Chromosomes, Fungal Cloning, Molecular Culture Media Escherichia coli - genetics Fundamental and applied biological sciences. Psychology Gene Library Genes, Fungal Genes. Genome Genetic Complementation Test Genotype Molecular and cellular biology Molecular genetics Mutation Nucleic Acid Hybridization Osmolar Concentration Phenotype Plasmids Restriction Mapping RNA, Messenger - genetics Saccharomyces cerevisiae Saccharomyces cerevisiae - genetics Saccharomyces cerevisiae - growth & development Transcription, Genetic Transformation, Genetic |
| Title | Cloning and characterization of a gene which determines osmotic stability in Saccharomyces cerevisiae |
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