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Modified Bimolecular Fluorescence Complementation Assay to Study the Inhibition of Transcription Complex Formation by JAZ Proteins

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Jasmonate Signaling

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1011))

Abstract

The jasmonate (JA) ZIM-domain (JAZ) proteins of Arabidopsis thaliana repress JA signaling and negatively regulate the JA responses. Recently, JAZ proteins have been found to inhibit the transcriptional function of several transcription factors, among which the basic helix-loop-helix (bHLH) (GLABRA3 [GL3], ENHANCER OF GLABRA3 [EGL3], and TRANSPARENT TESTA8 [TT8]) and R2R3-MYB (GL1 and MYB75) that can interact with each other to form bHLH–MYB complexes and further control gene expression. The bimolecular fluorescence complementation (BiFC) assay is a widely used technique to study protein–protein interactions in living cells. Here we describe a modified BiFC experimental procedure to study the inhibition of the formation of the bHLH (GL3)–MYB (GL1) complex by JAZ proteins.

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Acknowledgments

This research was supported by the National Science Foundation of China (91017012) and grants from the National Basic Research Program of China (973 Program 2011CB915404).

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Authors and Affiliations

  1. School of Life Sciences, Tsinghua University, Beijing, China

    Tiancong Qi, Susheng Song & Daoxin Xie

Authors
  1. Tiancong Qi
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  2. Susheng Song
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  3. Daoxin Xie
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Editor information

Editors and Affiliations

  1. Department of Plant Systems Biology, VIB, Technologiepark 927, Gent, 9052, Belgium

    Alain Goossens

  2. Department of Plant Systems Biology, VIB, Technologiepark 927, Gent, 9052, Belgium

    Laurens Pauwels

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Qi, T., Song, S., Xie, D. (2013). Modified Bimolecular Fluorescence Complementation Assay to Study the Inhibition of Transcription Complex Formation by JAZ Proteins. In: Goossens, A., Pauwels, L. (eds) Jasmonate Signaling. Methods in Molecular Biology, vol 1011. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-414-2_15

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  • DOI: https://doi.org/10.1007/978-1-62703-414-2_15

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-413-5

  • Online ISBN: 978-1-62703-414-2

  • eBook Packages: Springer Protocols

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