Skip to main content

Use of Transformation and Meiotic Gene Conversion to Construct a Yeast Strain Containing a Deletion in the Alcohol Dehydrogenase I Gene

  • Chapter
Book cover Genetic Engineering in Eukaryotes

Part of the book series: NATO Advanced Science Institutes Series ((NSSA,volume 61))

Abstract

Techniques have been developed using in vitro mutagenesis and yeast transformation to alter the genetic material of the yeast Saccharomyces cerevisiae (1). Features of the yeast system which make this possible include the following: the availability of selectable markers; the ease of transformation of yeast with exogenous DNA; the integration of DNA into yeast chromosomes by homologous recombination; and the ease of genetic manipulation of yeast cells. As an illustration of some of the features of the yeast system, we describe the construction of a yeast strain which contains a deletion of the 5′ end of the alcohol dehydrogenase I (ADHI) gene (ADC1) of yeast. Deletions are, in general, rare events in yeast so the ability to introduce in vitro created deletions into the yeast genome is quite useful.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Chapter
USD 29.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 84.99
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 109.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. S. Scherer and R. W. Davis, Replacement of chromosomal segments with altered DNA sequences constructed in vitro, Proc. Nat. Acad. Sci., USA 76:4951–4955(1979).

    Article  CAS  Google Scholar 

  2. M. Ciriacy, Genetics of alcohol dehydrogenase in Saccharomyces cerevisiae. I. Isolation and genetic analysis of adh mutants, Mutation Research 29:315–326(1975).

    Article  CAS  Google Scholar 

  3. M. Ciriacy, Genetics of alcohol dehydrogenase in Saccharomyces cerevisiae. II. Two loci controlling synthesis of the glucose repressible ADHII. Mol. Gen. Genet. 138:157–164(1975).

    Article  PubMed  CAS  Google Scholar 

  4. C. L. Denis, M. Ciriacy, and E. T. Young, A positive regulatory gene is required for accumulation of the functional messenger RNA for the glucose-repressible alcohol dehydrogenase from Saccharomyces cerevisiae, J. Mol. Biol. 148:355–368(1981).

    Article  PubMed  CAS  Google Scholar 

  5. V. M. Williamson, J. Bennetzen, E. T. Young, K. Nasmyth, and B. D. Hall, Isolation of the structural gene for alcohol dehydrogenase by genetic complementation in yeast. Nature 283:214–216(1980).

    Article  PubMed  CAS  Google Scholar 

  6. V. M. Williamson, E. T. Young, and M. Ciriacy, Transposable elements associated with constitutive expression of yeast alcohol dehydrogenase II, Cell 23:605–614(1981).

    Article  PubMed  CAS  Google Scholar 

  7. J. L. Bennetzen and B. D. Hall, The primary structure of the Saccharomyces cerevisiae gene for alcohol dehydrogenase I, J. Biol. Chem. 257:3018–3025(1982).

    PubMed  CAS  Google Scholar 

  8. D. W. Russell, V. M. Williamson, E. T. Young, and M. Smith, Nucleotide sequence of the yeast alcohol dehydrogenase II gene, manuscript in preparation.

    Google Scholar 

  9. M. Ciriacy, Isolation and characterization of further cis- and trans-acting regulatory elements involved in the synthesis of glucose-repressible alcohol dehydrogenase (ADHII), Mol. Gen. Genet. 176:427–431(1979).

    Article  CAS  Google Scholar 

  10. D. R. Beier and E. T. Young, Characterization of a regulatory region upstream of the ADR2 locus of S. cerevisiae, submitted.

    Google Scholar 

  11. H. L. Klein and T. D. Petes, Intrachromosomal gene conversion in yeast, Nature 289:144–148(1981).

    Article  PubMed  CAS  Google Scholar 

  12. J. A. Jackson and G. R. Fink, Gene conversion between duplicated genetic elements in yeast, Nature 292:306–311(1981).

    Article  PubMed  CAS  Google Scholar 

  13. T. L. Orr-Weaver, J. W. Szostak, and R. J. Rothstein, Yeast transformation: a model system for the study of recombination, Proc. Nat. Acad. Sci. USA 78:6354–6358(1981).

    Article  PubMed  CAS  Google Scholar 

  14. M. Ciriacy and V. M. Williamson, Analysis of mutations affecting Ty-mediated gene expression in Saccharomyces cerevisiae. Mol. Gen. Genet. 182:159–163(1981).

    Article  PubMed  CAS  Google Scholar 

  15. C. T. Korch, F. Lacroute, F. Exinger, A regulatory interaction between pyrimidine and purine biosynthesis via ureido succinic acid, Mol. Gen. Genet. 133:63–75(1974).

    Article  PubMed  CAS  Google Scholar 

  16. A. J. Berk and P. A. Sharp, Spliced early mRNAs of similar virus 40, Proc. Nat. Acad. Sci. U.S.A., 75:1274–1278(1978).

    Article  CAS  Google Scholar 

  17. V. M. Williamson, D. Cox, E. T. Young, D. W. Russell, and M. Smith, Characterization of transposable element-associated mutations that alter yeast alcohol dehydrogenase II expression, submitted.

    Google Scholar 

  18. K. A. Nasmyth, K. Tatchell, B. D. Hall, C. Astell, and M. Smith, Physical analysis of mating-type loci in Saccharomyces cerevisiae, Cold Spring Harbor Symp. Quant. Biol. 45:961–981(1980).

    Article  Google Scholar 

  19. J. D. Beggs, Transformation of yeast by a replicating hybrid Plasmid, Nature 275:104–108(1978).

    Article  PubMed  CAS  Google Scholar 

  20. A. Hinnen, J. B. Hicks, and G. R. Fink, Transformation of yeast, Proc. Nat. Acad. Sci. USA, 75:1929–1933(1978).

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 1983 Plenum Press, New York

About this chapter

Cite this chapter

Williamson, V.M., Beier, D., Young, E.T. (1983). Use of Transformation and Meiotic Gene Conversion to Construct a Yeast Strain Containing a Deletion in the Alcohol Dehydrogenase I Gene. In: Lurquin, P.F., Kleinhofs, A. (eds) Genetic Engineering in Eukaryotes. NATO Advanced Science Institutes Series, vol 61. Springer, Boston, MA. https://doi.org/10.1007/978-1-4684-4493-3_3

Download citation

  • DOI: https://doi.org/10.1007/978-1-4684-4493-3_3

  • Publisher Name: Springer, Boston, MA

  • Print ISBN: 978-1-4684-4495-7

  • Online ISBN: 978-1-4684-4493-3

  • eBook Packages: Springer Book Archive

Publish with us

Policies and ethics