Skip to main content

Protoplast Fusion in Saccharomyces cerevisiae

  • Protocol
Yeast Protocols

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 53))

Abstract

Saccharomyces cerevisiae can exist as either a haploid cell of either Mat a or Mat α mating type, a diploid cell or a polyploid cell, but the transfer of genetic material is normally restricted to conjugation between haploids of opposite mating type. It is possible, however, to isolate genetically stable yeast hybrids from strains that are unable to mate by using protoplast fusion. In this process, the yeast cell walls are enzymatically removed, the resulting protoplasts are fused together using polyethylene glycol (PEG) and calcium ions (Ca2+), before being embedded in agar under appropriate selective conditions. Colonies of genetically stable hybrids are normally visible within 5–10 d at 25°C.

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 74.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Maggio, B., Ahkong, Q. F., and Lucy, J. A. (1976) Poly(ethylene glycol) surface potential and cell fusion. Biochem. J. 158, 647–650.

    PubMed  CAS  Google Scholar 

  2. Papahadjopoulos, D., Vail, W. J., Pangborn, W. A., and Poste, G. (1976) Studies on membrane fusion II. Induction of fusion in pure phospholipid membranes by calcium ions and other divalent metals. Biochem. Biophys. Acta 448, 265–283.

    Article  PubMed  CAS  Google Scholar 

  3. Curran, B. P. G. and Carter, B. L. A. (1986) α-factor enhancement of hybrid formation by protoplast fusion in the yeast S. cerevisiae II. Curr. Genet. 10, 943–945.

    Article  PubMed  CAS  Google Scholar 

  4. Curran, B. P. G. and Carter, B. L. A. (1989) The α-factor enhancement of hybrid formation by protoplast fusion in S. cerevisiae can be mimicked by other procedures. Curr. Genet. 15, 303–305.

    Article  CAS  Google Scholar 

  5. Curran, B. P. G. and Carter, B. L. A. (1983) α-factor enhancement of hybrid formation by protoplast fusion in the yeast S. cerevisiae. J. Gen. Microbiol. 129, 1539–1591.

    Google Scholar 

  6. Bugeja, U. C., Whittaker, P. A., and Curran, B. P. G. (1988) The effect of α-factor on hybrid formation by protoplast fusion in S. cerevisiae. Fems. Microb. Letts. 51, 101–104.

    CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 1996 Humana Press Inc.,Totowa, NJ

About this protocol

Cite this protocol

Curran, B.P.G., Bugeja, V.C. (1996). Protoplast Fusion in Saccharomyces cerevisiae . In: Evans, I.H. (eds) Yeast Protocols. Methods in Molecular Biology™, vol 53. Humana Press. https://doi.org/10.1385/0-89603-319-8:45

Download citation

  • DOI: https://doi.org/10.1385/0-89603-319-8:45

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-319-1

  • Online ISBN: 978-1-59259-540-2

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics