DOI: 10.1126/science.274.5295.2069. Functional Analysis of the Genes of Yeast Chromosome V by Genetic Footprinting

Functional Analysis of the Genes of Yeast Chromosome V by Genetic Footprinting

10.1126/science.274.5295.2069

Crossref journal-article

American Association for the Advancement of Science (AAAS)

Science (221)

Abstract

Genetic footprinting was used to assess the phenotypic effects of Ty1 transposon insertions in 268 predicted genes of chromosome V of Saccharomyces cerevisiae. When seven selection protocols were used, Ty1 insertions in more than half the genes tested (157 of 268) were found to result in a detectable reduction in fitness. Results could not be obtained for fewer than 3 percent of the genes tested (7 of 268). Previously known mutant phenotypes were confirmed, and, for about 30 percent of the genes, new mutant phenotypes were identified.

Bibliography

Smith, V., Chou, K. N., Lashkari, D., Botstein, D., & Brown, P. O. (1996). Functional Analysis of the Genes of Yeast Chromosome V by Genetic Footprinting. Science, 274(5295), 2069â2074.

Authors 5

Victoria Smith (first)
Karen N. Chou (additional)
Deval Lashkari (additional)
David Botstein (additional)
Patrick O. Brown (additional)

References 28 Referenced 199

10.1016/0092-8674(86)90697-5
Oliver S. G., et al., Nature 357, 38 (1992). (10.1038/357038a0) / Nature by Oliver S. G. (1992)
10.1073/pnas.92.14.6479
Dietrich F., et al.., Nature, in press. In subsequent releases, additional telomeric sequences were added, making the full length of chromosome V 574,860 bp [updated in the Martinsried Institute for Protein Sequences (MIPS) Yeast Genome Database]. The number of predicted genes is derived from Dietrich et al. and from the MIPS database. Most of the possible additional and alternative open reading frames noted in the MIPS database were also covered by the primers used in this study. One to three independent gene-specific primers were used to analyze each gene. Primers were designed with the use of the program PRIMER (Whitehead Institute for Biomedical Research, Cambridge, MA) with a specified melting temperature of 69° to 73°C. Primers were synthesized by means of a 96-well array synthesizer [D. A. Lashkari, S. P. Hunicke-Smith, R. Norgren, R. W. Davis, T. Brennan, Proc. Natl. Acad. Sci. U.S.A.92, 7912 (1995)] and labeled at their 5′ end with 5-carboxyfluorescein (Applied Biosystems or Pharmacia). Approximately 85% of primers produced usable data. Alternative primers were synthesized for any gene for which the first primer failed to produce usable data. Each labeled gene-specific primer was used in a PCR with an unlabeled Ty1-specific primer, as described (2). / Nature by Dietrich F.
Brandriss M. C., J. Bacteriol. 138, 816 (1979). (10.1128/jb.138.3.816-822.1979) / J. Bacteriol. by Brandriss M. C. (1979)
Irie K., Yamaguchi K., Kawase K., Matsumoto K., Mol. Cell. Biol. 14, 3150 (1994). / Mol. Cell. Biol. by Irie K. (1994)
Graham L. A., Hill K. J., Stevens T. H., J. Biol. Chem. 270, 15037 (1995); (10.1074/jbc.270.25.15037) / J. Biol. Chem. by Graham L. A. (1995)
Abeijon C., et al., J. Cell Biol. 122, 307 (1993); (10.1083/jcb.122.2.307) / J. Cell Biol. by Abeijon C. (1993)
Prakash S., Prakash L., Genetics 87, 229 (1977). (10.1093/genetics/87.2.229) / Genetics by Prakash S. (1977)
Werner-Washburne M., Stone D. E., Craig E. A., Mol. Cell. Biol. 7, 2568 (1987). / Mol. Cell. Biol. by Werner-Washburne M. (1987)
10.1016/0092-8674(92)90447-K
Moehle C. M., Aynardi M. W., Kolodny M. R., Park F. J., Jones E. W., Genetics 115, 255 (1987); (10.1093/genetics/115.2.255) / Genetics by Moehle C. M. (1987)
Mitchell A. P., Magasanik B., Mol. Cell. Biol. 4, 2758 (1984). / Mol. Cell. Biol. by Mitchell A. P. (1984)
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10.1007/BF00337933
Masselot M., de Robichon-Szulmajster H., ibid. 139, 121 (1975); / ibid. by Masselot M. (1975)
Boonchird C., Messenguy F., Dubois E., ibid. 226, 154 (1991). / ibid. by Boonchird C. (1991)
Mitchell A. P., Magasanik B., Mol. Cell. Biol. 4, 2767 (1984). / Mol. Cell. Biol. by Mitchell A. P. (1984)
Chapman R. E., Munro S., EMBO J. 13, 4896 (1994). (10.1002/j.1460-2075.1994.tb06817.x) / EMBO J. by Chapman R. E. (1994)
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Nakafuku M., et al., ibid. 85, 1374 (1988). / ibid. by Nakafuku M. (1988)
Atkinson K. D., et al., J. Bacteriol. 141, 558 (1980). (10.1128/jb.141.2.558-564.1980) / J. Bacteriol. by Atkinson K. D. (1980)
Kim Y-J., Francisco L., Chen G.-C., Marcotte E., Chan C. S. M., J. Cell Biol. 127, 1381 (1994). (10.1083/jcb.127.5.1381) / J. Cell Biol. by Kim Y-J. (1994)

Dates

Type	When
Created	23 years ago (July 27, 2002, 5:45 a.m.)
Deposited	1 year ago (Aug. 7, 2024, 7:44 a.m.)
Indexed	4 months, 2 weeks ago (April 11, 2025, 5:06 a.m.)
Issued	28 years, 8 months ago (Dec. 20, 1996)
Published	28 years, 8 months ago (Dec. 20, 1996)
Published Print	28 years, 8 months ago (Dec. 20, 1996)

Funders 0

None

BibTeX

@article{Smith_1996, title={Functional Analysis of the Genes of Yeast Chromosome V by Genetic Footprinting}, volume={274}, ISSN={1095-9203}, url={http://dx.doi.org/10.1126/science.274.5295.2069}, DOI={10.1126/science.274.5295.2069}, number={5295}, journal={Science}, publisher={American Association for the Advancement of Science (AAAS)}, author={Smith, Victoria and Chou, Karen N. and Lashkari, Deval and Botstein, David and Brown, Patrick O.}, year={1996}, month=dec, pages={2069–2074} }

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