1887

Abstract

Summary: The gene of plays a key role in co-ordinating cell proliferation and nutrient availability. A 2·6 kb yeast DNA sequence has been cloned which fully suppresses the mutation. Integration of this sequence to demonstrate that the structural gene itself had been cloned proved difficult. Integration occurred only rarely and only at the locus which was also present on the integrating plasmid. To circumvent these difficulties an adjacent sequence, present on the original isolate from the gene library, was subcloned onto the integrating vector YIp5, after which directed integration proved straightforward. The integrated sequence was closely linked to , confirming that the structural gene had been cloned. A chromosomal restriction map of the region is presented; no gross changes were observed in the region as cells entered stationary phase.

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/content/journal/micro/10.1099/00221287-131-7-1797
1985-07-01
2024-04-25
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References

  1. Beggs J. D. 1978; Transformation of yeast by a replicating hybrid plasmid. Nature, London 275:104–108
    [Google Scholar]
  2. Beggs J. D. 1981; Gene cloning in yeast. In Genetic Engineering pp 175–203 Edited by Williamson W. London: Academic Press;
    [Google Scholar]
  3. Birnboim H. C., Doly J. 1979; A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Research 7:1513–1523
    [Google Scholar]
  4. Broach J. R. 1981; The yeast plasmid 2 µm circle. In The Molecular Biology of the Yeast Saccharomyces. Life Cycle and Inheritance pp 445–470 Edited by Strathern J. N., Jones E. W., Broach J. R. New York: Cold Spring Harbor Laboratory;
    [Google Scholar]
  5. Dawes I. W., Hardie I. H. 1974; Selective killing of vegetative cells in sporulated yeast cultures by exposure to diethyl ether. Molecular and General Genetics 80:281–289
    [Google Scholar]
  6. Deutch C. E., Parry J. M. 1974; Sphaeroplast formation in yeast during the transition from exponential phase to stationary phase. Journal of General Microbiology 80:259–268
    [Google Scholar]
  7. Dobson M. J., Kingsman S. M., Kingsman A. J. 1981; Sequence variation in the LEU2 region of Saccharomyces cerevisiae genome. Gene 16:133–139
    [Google Scholar]
  8. Hartwell L. H. 1974; Saccharomyces cerevisiae cell cycle. Bacteriological Reviews 38:164–198
    [Google Scholar]
  9. Hicks J. B. 1982; Cloning by complementation in yeast: the mating type genes. In Genetic Engineering vol 2 pp 219–248 Edited by Setlow J. K., Hollaender A. New York: Plenum Press;
    [Google Scholar]
  10. Johnston G. C., Ehrhardt C. W., Lorincz A., Carter B. L. A. 1979; Regulation of cell size in the yeast Saccharomyces cerevisiae . Journal of Bacteriology 137:1–5
    [Google Scholar]
  11. Lederberg E. M., Cohen S. N. 1974; Transformation of Salmonella typhimurium by plasmid deoxyribonucleic acid. Journal of Bacteriology 119:1072–1074
    [Google Scholar]
  12. Lillie S. H., Pringle J. R. 1980; Reserve carbohydrate metabolism in Saccharomyces cerevisiae: responses to nutrient limitation. Journal of Bacteriology 143:1384–1394
    [Google Scholar]
  13. Lorincz A., Carter B. L. A. 1979; Control of cell size at bud initiation in Saccharomyces cerevisiae . Journal of General Microbiology 113:287–296
    [Google Scholar]
  14. Maniatis T., Fritsch E. F., Sambrook J. 1982 Molecular Cloning: A Laboratory Manual New York: Cold Spring Harbor Laboratory.;
    [Google Scholar]
  15. Mortimer R. K., Schild D. 1981; Genetic mapping in Saccharomyces cerevisiae . In The Molecular Biology of the Yeast Saccharomyces. Life Cycle and Inheritance pp 11–26 Edited by Strathern J. N., Jones E. W., Broach J. R. New York: Cold Spring Harbor Laboratory:
    [Google Scholar]
  16. Nasmyth K. A., Tatchell K., Hall B. D., Astell C., Smith M. 1981; A physical analysis of mating type loci in Saccharomyces cerevisiae . Cold Spring Harbor Symposia on Quantitative Biology 45:961–975
    [Google Scholar]
  17. Orr-Weaver T. L., Szostak J. W., Rothstein R. J. 1981; Yeast transformation: a model system for the study of recombination. Proceedings of the National Academy of Sciences of the United States of America 78:6354–6358
    [Google Scholar]
  18. Orr-Weaver T. L., Szostak J. W., Rothstein R. J. 1983; Genetic applications of yeast transformation with linear and gapped plasmids. Methods in Enzymology 101:228–245
    [Google Scholar]
  19. Parry J. M., Davies P. J., Evans W. E. 1976; The effect of “cell age” upon the lethal effects of physical and chemical mutagens in yeast. Molecular and General Genetics 146:27–35
    [Google Scholar]
  20. Saul D. J., Walton E. F., Sudbury P. E., Carter B. L. A. 1985; Saccharomyces cerevisiae whi2 mutants in stationary phase retain the properties of exponentially growing cells. Journal of General Microbiology 131: (in the Press)
    [Google Scholar]
  21. Schenberg-Frascino A., Moustacchi E. 1972; Lethal and mutagenic effects of elevated temperature on haploid yeast. 1. Variations in sensitivity during the cell cycle. Molecular and General Genetics 115:243–257
    [Google Scholar]
  22. Sherman F., Fink G. R., Lawrence G. W. 1974 Methods in Yeast Genetics New York: Cold Spring Harbor Laboratory.;
    [Google Scholar]
  23. Snow R. 1966; An enrichment method for auxotrophic yeast mutants using “Nystatin”. Nature, London 211:206–207
    [Google Scholar]
  24. Southern E. 1975; Detection of specific sequences among DNA fragments separated by gene electrophoresis. Journal of Molecular Biology 98:503–517
    [Google Scholar]
  25. Struhl K., Stinchcomb D. T., Scherer S., Davis R. W. 1979; High frequency transformation in yeast: autonomous replication of hybrid DNA molecules. Proceedings of the National Academy of Sciences of the United States of America 76:1035–1039
    [Google Scholar]
  26. Sudbery P. E., Goodey A. R., Carter B. L. A. 1980; Genes which control cell proliferation in the yeast Saccharomyces cerevisiae . Nature, London 288:401–404
    [Google Scholar]
  27. Twigg A. J., Sherratt D. 1980; Trans-complementable copy-number mutants of plasmid ColE1. Nature, London 283:216–218
    [Google Scholar]
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