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Plant Hormones pp 271-281 | Cite as

Characterizing Auxin Response Circuits in Saccharomyces cerevisiae by Flow Cytometry

  • Edith Pierre-Jerome
  • R. Clay Wright
  • Jennifer L. NemhauserEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1497)

Abstract

Recapitulation of the nuclear auxin response pathway in Saccharomyces cerevisiae (yeast) provides a means to functionally assay the contribution of individual signaling components to response dynamics. Here, we describe a time course assay for characterizing auxin response circuits using flow cytometry. This method allows for quantitative measurements of the dynamic response of up to 12 circuits (strains) at once. We also describe a steady-state assay and how to utilize an R package we developed to facilitate data analysis.

Key words

Synthetic biology Budding yeast Fluorescent reporters Signaling dynamics 

References

  1. 1.
    Pierre-Jerome E, Jang SS, Havens KA, Nemhauser JL, Klavins E (2014) Recapitulation of the forward nuclear auxin response pathway in yeast. Proc Natl Acad Sci U S A 111(26):9407–9412Google Scholar
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    Havens KA, Guseman JM, Jang SS et al. (2012) A synthetic approach reveals extensive tunability of auxin signaling. Plant Physiol 160:135–142Google Scholar
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    R Core Team (2015) R: a language and environment for statistical computing. R Foundation for Statistical Computing, Vienna, Austria, http://www.R-project.org/
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    Ellis B, Haaland P, Hahne F, Le Meur N, Gopalakrishnan N, Spidlen J, Jiang M flowCore: basic structures for flow cytometry data. R package version 1.36.9. https://www.bioconductor.org/packages/release/bioc/html/flowCore.html
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    Baxter SK, Lambert AR, Scharenberg AM, Jarjour J (2013) Flow cytometric assays for interrogating LAGLIDADG homing endonuclease DNAbinding and cleavage properties. Methods Mol Biol 978:45–61Google Scholar

Copyright information

© Springer Science+Business Media New York 2017

Authors and Affiliations

  • Edith Pierre-Jerome
    • 1
  • R. Clay Wright
    • 1
  • Jennifer L. Nemhauser
    • 1
    Email author
  1. 1.Department of BiologyUniversity of WashingtonSeattleUSA

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