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Research Article

An Improved Cerulean Fluorescent Protein with Enhanced Brightness and Reduced Reversible Photoswitching

  • Michele L. Markwardt,

    Affiliation: Department of Physiology, University of Maryland School of Medicine, Baltimore, Maryland, United States of America

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  • Gert-Jan Kremers,

    Affiliation: Department of Molecular Physiology and Biophysics, Vanderbilt University, Nashville, Tennessee, United States of America

    Current address: Department of Cell Biology, Erasmus Medical Center, Rotterdam, The Netherlands

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  • Catherine A. Kraft,

    Affiliation: Department of Physiology, University of Maryland School of Medicine, Baltimore, Maryland, United States of America

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  • Krishanu Ray,

    Affiliation: Center for Fluorescence Spectroscopy and the Department of Biochemistry and Molecular Biology, University of Maryland School of Medicine, Baltimore, Maryland, United States of America

    X
  • Paula J. C. Cranfill,

    Affiliation: National High Magnetic Field Laboratory and Department of Biological Science, The Florida State University, Tallahassee, Florida, United States of America

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  • Korey A. Wilson,

    Affiliation: National High Magnetic Field Laboratory and Department of Biological Science, The Florida State University, Tallahassee, Florida, United States of America

    X
  • Richard N. Day,

    Affiliation: Department of Cellular and Integrative Physiology, Indiana University School of Medicine, Indianapolis, Indiana, United States of America

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  • Rebekka M. Wachter,

    Affiliation: Department of Chemistry and Biochemistry, Arizona State University, Tempe, Arizona United States of America

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  • Michael W. Davidson,

    Affiliation: National High Magnetic Field Laboratory and Department of Biological Science, The Florida State University, Tallahassee, Florida, United States of America

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  • Mark A. Rizzo mail

    mrizz001@umaryland.edu

    Affiliation: Department of Physiology, University of Maryland School of Medicine, Baltimore, Maryland, United States of America

    X
  • Published: March 29, 2011
  • DOI: 10.1371/journal.pone.0017896

About the Authors

Michele L. Markwardt, Catherine A. Kraft, Mark A. Rizzo
Department of Physiology, University of Maryland School of Medicine, Baltimore, Maryland, United States of America
Gert-Jan Kremers
Department of Molecular Physiology and Biophysics, Vanderbilt University, Nashville, Tennessee, United States of America
Krishanu Ray
Center for Fluorescence Spectroscopy and the Department of Biochemistry and Molecular Biology, University of Maryland School of Medicine, Baltimore, Maryland, United States of America
Paula J. C. Cranfill, Korey A. Wilson, Michael W. Davidson
National High Magnetic Field Laboratory and Department of Biological Science, The Florida State University, Tallahassee, Florida, United States of America
Richard N. Day
Department of Cellular and Integrative Physiology, Indiana University School of Medicine, Indianapolis, Indiana, United States of America
Rebekka M. Wachter
Department of Chemistry and Biochemistry, Arizona State University, Tempe, Arizona United States of America

Corresponding Author

Email: mrizz001@umaryland.edu

Competing Interests

The mutant CFPs described in this article are the topic of a pending patent application from the University of Maryland, Baltimore titled "Fluorescent Proteins and Uses Thereof" (SN 61/249,712). This patent covers the mutations used to derive mCerulean2 and mCerulean2.N variants that are the precursors to mCerulean3. Although the authors are pursuing commercial licensing and sale of their CFP reagents through companies like Clontech and Life Technologies, this does not alter their acceptance and adherence to the PLoS ONE policy as well as National Institutes of Health (NIH) policy for reagent sharing. All reagents described in the article are freely available upon reasonable request for the purpose of academic, non-commercial research, which will likely include deposition of the plasmids encoding mCerulean3 in a repository such as addgene.org.

Author Contributions

Conceived and designed the experiments: MLM GJK RND RMW MWD MAR. Performed the experiments: MLM GJK CAK KR PJCC KAW RND MWD MAR. Analyzed the data: MLM GJK CAK KR PJCC KAW RND MWD MAR. Contributed reagents/materials/analysis tools: RMW. Wrote the paper: GJK CAK RND RMW MWD MAR.