Abstract
Transient expression assays are valuable techniques to study in vivo the transcriptional regulation of gene expression. These methods allow to assess the transcriptional properties of a given transcription factor (TF) or a complex of regulatory proteins against specific DNA motifs, called cis-regulatory elements. Here, we describe a fast, efficient, and reliable method based on the use of Physcomitrella patens protoplasts that allows the study of gene expression in a qualitative and quantitative manner by combining the advantage of GFP (green fluorescent protein) as a marker of promoter activity with flow cytometry for accurate measurement of fluorescence in individual cells.
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Acknowledgement
We thank the “Plateforme de cytologie et imagerie végétale (PCIV)” of the Plant Observatory from the Institut Jean-Pierre Bourgin (IJPB) for excellent technical support. We also thank F. Charlot and F. Nogué (IJPB) for their help and advice in setting up this protocol and for providing P. patens spores. We acknowledge China Scholarship Council (CSC) for supporting W.X. This work was supported by two projects, STREG (Plant-KBBE) and CERES (ANR-2010-BLAN-1238).
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Thévenin, J., Xu, W., Vaisman, L., Lepiniec, L., Dubreucq, B., Dubos, C. (2016). The Physcomitrella patens System for Transient Gene Expression Assays. In: Hehl, R. (eds) Plant Synthetic Promoters. Methods in Molecular Biology, vol 1482. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6396-6_10
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DOI: https://doi.org/10.1007/978-1-4939-6396-6_10
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