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Chemical Biology pp 63–73Cite as

Small-Molecule High-Throughput Screening Utilizing Xenopus Egg Extract

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1263))

Abstract

Screens for small-molecule modulators of biological pathways typically utilize cultured cell lines, purified proteins, or, recently, model organisms (e.g., zebrafish, Drosophila, C. elegans). Herein, we describe a method for using Xenopus laevis egg extract, a biologically active and highly tractable cell-free system that recapitulates a legion of complex chemical reactions found in intact cells. Specifically, we focus on the use of a luciferase-based fusion system to identify small-molecule modulators that affect protein turnover.

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Acknowledgments

We thank Laurie Lee for critical reading of the manuscript. M.R.B. is supported by a National Cancer Institute training grant (T32 CA119925). E.L. is supported by the National Institutes of Health (R01GM081635 and R01GM103926). R.Y. is supported by Award Number 8UL1TR000149 from the National Center for Advancing Translational Sciences and the CTRC P30 Cancer Center Support Grant from the National Cancer Institute (CA054174). S.R.H. is supported by the National Cancer Institute (P50 CA095103).

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Authors and Affiliations

  1. Department of Cell and Developmental Biology, Vanderbilt University Medical Center, 465 21st Avenue South, U-4213A Learned Lab/MRBIII, Nashville, TN, 37232-8240, USA

    Matthew R. Broadus, Stephen R. Hann & Ethan Lee

  2. Department of Molecular Medicine, Institute of Biotechnology, The University of Texas Health Science Center at San Antonio, San Antonio, TX, USA

    P. Renee Yew

  3. Vanderbilt Institute of Chemical Biology, Vanderbilt University Medical Center, Nashville, TN, USA

    Ethan Lee

  4. Vanderbilt-Ingram Cancer Center, Vanderbilt University Medical Center, Nashville, TN, USA

    Ethan Lee

Authors
  1. Matthew R. Broadus
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  2. P. Renee Yew
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  3. Stephen R. Hann
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  4. Ethan Lee
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Corresponding author

Correspondence to Ethan Lee .

Editor information

Editors and Affiliations

  1. Department of Medicine - Cardiovascular Medicine, Vanderbilt Institute of Chemical Biology, Vanderbilt University School of Medicine, Nashville, Tennessee, USA

    Jonathan E. Hempel

  2. Department of Cell and Developmental Biology, Vanderbilt Institute of Chemical Biology, Vanderbilt University School of Medicine, Nashville, Tennessee, USA

    Charles H. Williams

  3. Department of Medicine - Cardiovascular Medicine, Department of Pharmacology, Department of Cell and Developmental Biology, Vanderbilt Institute of Chemical Biology, Vanderbilt University School of Medicine, Nashville, Tennessee, USA

    Charles C. Hong

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Broadus, M.R., Yew, P.R., Hann, S.R., Lee, E. (2015). Small-Molecule High-Throughput Screening Utilizing Xenopus Egg Extract. In: Hempel, J., Williams, C., Hong, C. (eds) Chemical Biology. Methods in Molecular Biology, vol 1263. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2269-7_5

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  • DOI: https://doi.org/10.1007/978-1-4939-2269-7_5

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-2268-0

  • Online ISBN: 978-1-4939-2269-7

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