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Reporter Gene Assays pp 135–148Cite as

Using YFP as a Reporter of Gene Expression in the Green Alga Chlamydomonas reinhardtii

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1755))

Abstract

The unicellular green alga Chlamydomonas reinhardtii is a valuable experimental system in plant biology for studying metal homeostasis. Analyzing transcriptional regulation with promoter-fusion constructs in C. reinhardtii is a powerful method for connecting metal-responsive regulation with cis-regulatory elements, but overcoming expression-level variability between transformants and optimizing experimental conditions can be laborious. Here, we provide detailed protocols for the high-throughput cultivation of C. reinhardtii and assaying Venus fluorescence as a reporter for promoter activity. We also describe procedural considerations for relating metal supply to transcriptional activity.

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Acknowledgments

This work was supported by the Division of Chemical Sciences, Geosciences, and Biosciences, Office of Basic Energy Sciences of the US Department of Energy (DE-FD02-04ER15529) and the National Institutes of Health (NIH) R24 GM42143 to SM, and the Office of Biological and Environmental Research of the United States Department of Energy (CEB-H). We are grateful to Prof. Ralph Bock for providing C. reinhardtii UVM11 and pJR39, Dr. Ian Blaby for critical reading of the manuscript, and Britany Reddish for technical support during protocol development.

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Authors and Affiliations

  1. Department of Biology, Brookhaven National Laboratory, Upton, NY, USA

    Crysten E. Blaby-Haas

  2. Department of Chemistry and Biochemistry, University of California Los Angeles, Los Angeles, CA, USA

    M. Dudley Page & Sabeeha S. Merchant

  3. Institute for Genomics and Proteomics, University of California Los Angeles, Los Angeles, CA, USA

    Sabeeha S. Merchant

Authors
  1. Crysten E. Blaby-Haas
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  2. M. Dudley Page
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  3. Sabeeha S. Merchant
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Corresponding author

Correspondence to Crysten E. Blaby-Haas .

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Editors and Affiliations

  1. Department of Molecular and Medical Pharmacology, David Geffen School of Medicine, University of California Los Angeles, Los Angeles, California, USA

    Robert Damoiseaux

  2. Pfizer, Inc., Cambridge, Massachusetts, USA

    Samuel Hasson

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Blaby-Haas, C.E., Page, M.D., Merchant, S.S. (2018). Using YFP as a Reporter of Gene Expression in the Green Alga Chlamydomonas reinhardtii . In: Damoiseaux, R., Hasson, S. (eds) Reporter Gene Assays. Methods in Molecular Biology, vol 1755. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7724-6_10

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  • DOI: https://doi.org/10.1007/978-1-4939-7724-6_10

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7722-2

  • Online ISBN: 978-1-4939-7724-6

  • eBook Packages: Springer Protocols

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