Skip to main content

Transformation of Intact Cells of Saccharomyces cerevisiae: Lithium Methods and Possible Underlying Mechanism

  • Chapter
  • First Online:
Genetic Transformation Systems in Fungi, Volume 1

Part of the book series: Fungal Biology ((FUNGBIO))

  • 2810 Accesses

Abstract

Since it was initially established, the lithium method for transforming Saccharomyces cerevisiae intact cells has been modified, and a possible underlying mechanism has been elucidated. This method requires polyethylene glycol (PEG) and transformation efficiency is enhanced by lithium and single-stranded carrier DNA (ssDNA). Here, we describe the original lithium method, the modified method, and the possible underlying mechanism, in which plasmid DNA that was absorbed onto the cell wall enters into the cells via endocytotic membrane invagination and lithium and ssDNA synergistically alter cell wall structure and enhance transformation efficiency.

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 129.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 169.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 169.99
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

  • Gietz RD, Woods RA (2002) Transformation of yeast by lithium acetate/single-stranded carrier DNA/polyethylene glycol method. Methods Enzymol 350:87–96

    Article  PubMed  CAS  Google Scholar 

  • Gietz D, St Jean A, Woods RA, Schiestl RH (1992) Improved method for high efficiency transformation of intact yeast cells. Nucleic Acids Res 20:1425

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  • Gietz RD, Schiestl RH, Willems AR, Woods RA (1995) Studies on the transformation of intact yeast cells by the LiAc/SS-DNA/PEG procedure. Yeast 11:355–360

    Article  PubMed  CAS  Google Scholar 

  • Gurrieri S, Wells KS, Johnson ID, Bustamante C (1997) Direct visualization of individual DNA molecules by fluorescence microscopy: characterization of the factors affecting signal/background and optimization of imaging conditions using YOYO. Anal Biochem 249:44–53

    Article  PubMed  CAS  Google Scholar 

  • Hayama Y, Fukuda Y, Kawai S, Hashimoto W, Murata K (2002) Extremely simple, rapid and highly efficient transformation method for the yeast Saccharomyces cerevisiae using glutathione and early log phase cells. J Biosci Bioeng 94:166–171

    Article  PubMed  CAS  Google Scholar 

  • Hinnen A, Hicks JB, Fink GR (1978) Transformation of yeast. Proc Natl Acad Sci U S A 75:1929–1933

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  • Ito H, Fukuda Y, Murata K, Kimura A (1983) Transformation of intact yeast cells treated with alkali cations. J Bacteriol 153:163–168

    PubMed  CAS  PubMed Central  Google Scholar 

  • Kawai S, Pham TA, Nguyen HT, Nankai H, Utsumi T, Fukuda Y, Murata K (2004) Molecular insights on DNA delivery into Saccharomyces cerevisiae. Biochem Biophys Res Commun 317:100–107

    Article  PubMed  CAS  Google Scholar 

  • Kawai S, Hashimoto W, Murata K (2010) Transformation of Saccharomyces cerevisiae and other fungi: methods and possible underlying mechanism. Bioeng Bugs 1:395–403

    Article  PubMed  PubMed Central  Google Scholar 

  • Pham TA, Kawai S, Kono E, Murata K (2011a) The role of cell wall revealed by the visualization of Saccharomyces cerevisiae transformation. Curr Microbiol 62:956–961

    Article  PubMed  CAS  Google Scholar 

  • Pham TA, Kawai S, Murata K (2011b) Visualization of the synergistic effect of lithium acetate and single-stranded carrier DNA on Saccharomyces cerevisiae transformation. Curr Genet 57:233–239

    Article  PubMed  CAS  Google Scholar 

  • Schiestl RH, Gietz RD (1989) High efficiency transformation of intact yeast cells using single stranded nucleic acids as a carrier. Curr Genet 16:339–346

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Kousaku Murata Ph.D. .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2015 Springer International Publishing Switzerland

About this chapter

Cite this chapter

Kawai, S., Murata, K. (2015). Transformation of Intact Cells of Saccharomyces cerevisiae: Lithium Methods and Possible Underlying Mechanism. In: van den Berg, M., Maruthachalam, K. (eds) Genetic Transformation Systems in Fungi, Volume 1. Fungal Biology. Springer, Cham. https://doi.org/10.1007/978-3-319-10142-2_18

Download citation

Publish with us

Policies and ethics