Skip to main content

Markers, Selection, and Media in Yeast Artificial Chromosome Cloning

  • Protocol
YAC Protocols

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 349))

  • 834 Accesses

Abstract

Yeast have proven to be a valuable and flexible host for cloning and manipulating large fragments of foreign genetic material, providing valuable reagents for the study of gene function and regulation in a variety of applications. However, this approach requires a familiarity with basic procedures derived from yeast genetics, and lack of experience can be a barrier to researchers with backgrounds limited to bacterial and mammalian culture systems. To address this issue, this chapter introduces and discusses the genetic markers and selection systems used in the genetic manipulation of yeast, with particular reference to yeast artificial chromosome (YAC) cloning and manipulation. Detailed descriptions of culture media are provided, allowing for the growth and selection of standard YAC strains, as well as for specialist applications.

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

Access this chapter

Protocol
USD 49.95
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 EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 109.00
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 109.99
Price excludes VAT (USA)
  • Durable hardcover 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

References

  1. Bernstein, H. (1961) Imidazole compounds accumulated by purine mutants of Neurospora crassa. J. Gen. Microbiol. 25, 41–46.

    CAS  Google Scholar 

  2. Burke, D. T., Carle, G. F., and Olson, M. V. (1987) Cloning of large segments of exogenous DNA into yeast by means of artificial chromosome vectors. Science 236, 806–812.

    Article  CAS  PubMed  Google Scholar 

  3. Grenson, M., Mousset, M., Wiame, J. M., and Bechet, J. (1966) Multiplicity of the amino acid permeases in Saccharomyces cerevisiae, I. Evidence for a specific arginine-transporting system. Biochim. Biophys. Acta. 127, 325–338.

    CAS  PubMed  Google Scholar 

  4. Whelan, W. L., Gocke. E., and Manney, T. R. (1979) The CAN1 locus of Saccharomyces cerevisiae: fine structure analysis and forward mutation rates. Genetics 91, 35–51.

    CAS  PubMed  Google Scholar 

  5. Boeke, J. D., LaCroute, F., and Fink, G. R. (1984) A positive selection for mutants lacking orotidine-5′-phosphate decarboxylase activity in yeast: 5-fluoro-orotic acid resistance. Mol. Gen. Genet. 197, 345–346.

    Article  CAS  PubMed  Google Scholar 

  6. Heikoop, J. C., Steensma, Y., van Ommen, G.-J. B., and den Dunnen, J. T. (1994) A simple and rapid method for separating co-cloned YACs. Trends Genet. 10, 40.

    Article  CAS  PubMed  Google Scholar 

  7. Chattoo, B. B., Sherman, F., Azubalis, D. A., Fjellstedt, T. A., Mehnert, D., and Ogur, M. (1979) Selection of lys2 mutants of the yeast Saccharomyces cerevisiae by the utilization of α-amino adipate. Genetics 93, 51–65.

    CAS  PubMed  Google Scholar 

  8. Smith, D. R., Smyth, A. P., and Moir, D. T. (1990) Amplification of large artificial chromosomes. Proc. Natl. Acad. Sci. USA 87, 8242–8246.

    Article  CAS  PubMed  Google Scholar 

  9. Larionov, V., Kouprina, N., Nikolaishvili, N., and Resnick, M. A. (1994) Recombination during transformation as a source of chimeric mammalian artificial chromosomes in yeast (YACs). Nucl. Acids Res. 22, 4154–4162.

    Article  CAS  PubMed  Google Scholar 

  10. Kouprina, N., Eldarov, M., Moyzis, R., Resnick, M., and Larionov, V. (1994) A model system to assess the integrity of mammalian YACs during transformation and propagation in yeast. Genomics 21, 7–17.

    Article  CAS  PubMed  Google Scholar 

  11. Haldi, M., Perrot, V., Saumier, M., et al. (1994) Large human YACs constructed in a rad52 strain show a reduced rate of chimerism. Genomics 24, 478–484.

    Article  CAS  PubMed  Google Scholar 

  12. McCusker, J. H. and Davis, R. W. (1991) The use of proline as a nitrogen source causes hypersensitivity to, and allows more economical use of 5FOA in Saccharomyces cerevisiae. Yeast 7, 607–608.

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2006 Humana Press Inc., Totowa NJ

About this protocol

Cite this protocol

Markie, D. (2006). Markers, Selection, and Media in Yeast Artificial Chromosome Cloning. In: MacKenzie, A. (eds) YAC Protocols. Methods in Molecular Biology™, vol 349. Humana Press. https://doi.org/10.1385/1-59745-158-4:1

Download citation

  • DOI: https://doi.org/10.1385/1-59745-158-4:1

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-612-2

  • Online ISBN: 978-1-59745-158-1

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics