Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Brief Communication
  • Published:

Plasmid-chromosome shuffling for non-deletion alleles in yeast

Nature Methods volume 5, pages 167–169 (2008)Cite this article

Abstract

Here we describe a facile plasmid-chromosome shuffling technique for generating and analyzing non-deletion alleles in the yeast Saccharomyces cerevisiae. This technique takes advantage of an existing set of genome-wide haploid-convertible heterozygous diploid yeast knockout mutants. This simple method will facilitate characterization of essential gene functions and genome-wide investigation of protein structure-function relationships.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1: A schematic of random mutagenesis with plasmid-chromosome shuffling.
Figure 2: Isolation and characterization of MMS-hypersensitive pol30 alleles using plasmid-chromosome shuffling.

Similar content being viewed by others

References

  1. Boeke, J.D., Trueheart, J., Natsoulis, G. & Fink, G.R. Methods Enzymol. 154, 164–175 (1987).

    Article  CAS  PubMed  Google Scholar 

  2. Budd, M. & Campbell, J.L. Proc. Natl. Acad. Sci. USA 84, 2838–2842 (1987).

    Article  CAS  PubMed  Google Scholar 

  3. Muhlrad, D., Hunter, R. & Parker, R. Yeast 8, 79–82 (1992).

    Article  CAS  PubMed  Google Scholar 

  4. Pan, X. et al. Cell 124, 1069–1081 (2006).

    Article  CAS  PubMed  Google Scholar 

  5. Pan, X. et al. Mol. Cell 16, 487–496 (2004).

    Article  CAS  PubMed  Google Scholar 

  6. Tong, A.H. et al. Science 294, 2364–2368 (2001).

    Article  CAS  PubMed  Google Scholar 

  7. Giaever, G. et al. Nature 418, 387–391 (2002).

    Article  CAS  PubMed  Google Scholar 

  8. Amin, N.S. & Holm, C. Genetics 144, 479–493 (1996).

    CAS  PubMed  PubMed Central  Google Scholar 

  9. Hoege, C., Pfander, B., Moldovan, G.L., Pyrowolakis, G. & Jentsch, S. Nature 419, 135–141 (2002).

    Article  CAS  PubMed  Google Scholar 

  10. Lau, P.J., Flores-Rozas, H. & Kolodner, R.D. Mol. Cell. Biol. 22, 6669–6680 (2002).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Krishna, T.S., Kong, X.P., Gary, S., Burgers, P.M. & Kuriyan, J. Cell 79, 1233–1243 (1994).

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We thank S. Kron (University of Chicago) for advice in implementing the 96-well yeast plasmid recovery protocol. This work was supported by a Baylor College of Medicine seed grant to X.P. and US National Institutes of Health (NIH) Roadmap grant “Technology Center for Networks and Pathways” (RR020839) to J.D.B. R.S.S. is supported by NIH P01 grant HD39691.

Author information

Author notes

  1. Zhiwei Huang and Richard S Sucgang: These authors contributed equally to this work.

Authors and Affiliations

  1. Verna and Marrs McLean Department of Biochemistry and Molecular Biology, Baylor College of Medicine, One Baylor Plaza, Houston, 77030, Texas, USA

    Zhiwei Huang, Richard S Sucgang, Xiaomin Shi & Xuewen Pan

  2. Department of Molecular and Human Genetics, Baylor College of Medicine, One Baylor Plaza, Houston, 77030, Texas, USA

    Xuewen Pan

  3. High Throughput Biology Center, The Johns Hopkins University School of Medicine, 733 N. Broadway, Baltimore, 21205, Maryland, USA

    Yu-yi Lin & Jef D Boeke

Authors
  1. Zhiwei Huang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Richard S Sucgang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Yu-yi Lin
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Xiaomin Shi
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Jef D Boeke
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Xuewen Pan
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Jef D Boeke or Xuewen Pan.

Supplementary information

Supplementary Text and Figures

Supplementary Tables 1–6, Supplementary Methods (PDF 398 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Huang, Z., Sucgang, R., Lin, Yy. et al. Plasmid-chromosome shuffling for non-deletion alleles in yeast. Nat Methods 5, 167–169 (2008). https://doi.org/10.1038/nmeth.1173

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nmeth.1173

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing