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Expression of Epitope-Tagged Proteins in Arabidopsis Leaf Mesophyll Protoplasts

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Immunoelectron Microscopy

Part of the book series: Methods in Molecular Biology ((MIMB,volume 657))

Abstract

Advances in genomic and proteomic platforms enable high-throughput studies for regulatory factors and interactors involved in signaling network at a molecular level. However, it has never been trivial to verify the omics data in vivo or functionally integrate the data in a cell signaling context. For plants, genetic approaches using knockout mutants and transgenic lines have been mainly used to characterize functions of gene products in vivo. In general, such approaches demand a longer time and a higher cost and have difficulties in understanding gene functions comprehensively in a high-throughput manner. Transient gene expression is a method of choice to examine the cellular functions of genetic components in vivo. The leaf mesophyll protoplasts (LMP) provide a perfect system to transiently express a gene encoding an epitope-tagged protein of interest and quickly and reliably trace subcellular locales of the protein in a near high-throughput manner. Here, a simple and straightforward method for isolating leaf meshophyll protoplasts from Arabidopsis has been described in detail to help beginners initiate their first cell-based functional genomic analysis.

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Acknowledgments

We thank Dr. Donald Hunter for his help in the preparation of this chapter. This work was supported by National Research Foundation of Korea Grant 2010-0007068 to Y.-H. Cho and grants 2009-0085565 and 2010-0016989 to S.-D. Yoo.

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Cho, YH., Yoo, SD. (2010). Expression of Epitope-Tagged Proteins in Arabidopsis Leaf Mesophyll Protoplasts. In: Schwartzbach, S., Osafune, T. (eds) Immunoelectron Microscopy. Methods in Molecular Biology, vol 657. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60761-783-9_3

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  • DOI: https://doi.org/10.1007/978-1-60761-783-9_3

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-60761-782-2

  • Online ISBN: 978-1-60761-783-9

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