Arabidopsis mesophyll protoplasts

A versatile cell system for transient gene expression analysis

Sang-Dong Yoo, Young Hee Cho, Jen Sheen

Research output: Contribution to journalArticle

1900 Citations (Scopus)

Abstract

The transient gene expression system using Arabidopsis mesophyll protoplasts has proven an important and versatile tool for conducting cell-based experiments using molecular, cellular, biochemical, genetic, genomic and proteomic approaches to analyze the functions of diverse signaling pathways and cellular machineries. A well-established protocol that has been extensively tested and applied in numerous experiments is presented here. The method includes protoplast isolation, PEG-calcium transfection of plasmid DNA and protoplast culture. Physiological responses and high-throughput capability enable facile and cost-effective explorations as well as hypothesis-driven tests. The protoplast isolation and DNA transfection procedures take 6-8 h, and the results can be obtained in 2-24 h. The cell system offers reliable guidelines for further comprehensive analysis of complex regulatory mechanisms in whole-plant physiology, immunity, growth and development.

Original languageEnglish
Pages (from-to)1565-1572
Number of pages8
JournalNature Protocols
Volume2
Issue number7
DOIs
Publication statusPublished - 2007 Jul 1
Externally publishedYes

Fingerprint

Protoplasts
Arabidopsis
Gene expression
Gene Expression
DNA
Physiology
Polyethylene glycols
Transfection
Plasmids
Plant Immunity
Experiments
Throughput
Plant Physiological Phenomena
Calcium
Plant Development
Growth and Development
Proteomics
Costs
Molecular Biology
Guidelines

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Arabidopsis mesophyll protoplasts : A versatile cell system for transient gene expression analysis. / Yoo, Sang-Dong; Cho, Young Hee; Sheen, Jen.

In: Nature Protocols, Vol. 2, No. 7, 01.07.2007, p. 1565-1572.

Research output: Contribution to journalArticle

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