Expression of epitope-tagged proteins in arabidopsis leaf mesophyll protoplasts

Young Hee Cho, Sang Dong Yoo

Research output: Chapter in Book/Report/Conference proceedingChapter

7 Citations (Scopus)

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.

Original languageEnglish
Title of host publicationImmunoelectron Microscopy
Subtitle of host publicationMethods and Protocols
EditorsSteven Schwartzbach, Tetsuaki Osafune
Pages33-42
Number of pages10
DOIs
Publication statusPublished - 2010

Publication series

NameMethods in Molecular Biology
Volume657
ISSN (Print)1064-3745

Keywords

  • Arabidopsis
  • Epitope tag
  • High-throughput
  • Leaf mesophyll protoplasts
  • Transient expression

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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  • Cite this

    Cho, Y. H., & Yoo, S. D. (2010). Expression of epitope-tagged proteins in arabidopsis leaf mesophyll protoplasts. In S. Schwartzbach, & T. Osafune (Eds.), Immunoelectron Microscopy: Methods and Protocols (pp. 33-42). (Methods in Molecular Biology; Vol. 657). https://doi.org/10.1007/978-1-60761-783-9_3