Arabidopsis mesophyll protoplasts: A versatile cell system for transient gene expression analysis

Sang Dong Yoo; Young Hee Cho; Jen Sheen

doi:10.1038/nprot.2007.199

Arabidopsis mesophyll protoplasts: A versatile cell system for transient gene expression analysis

Sang Dong Yoo, Young Hee Cho, Jen Sheen

Department of Life Sciences

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2637 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 language	English
Pages (from-to)	1565-1572
Number of pages	8
Journal	Nature Protocols
Volume	2
Issue number	7
DOIs	https://doi.org/10.1038/nprot.2007.199
Publication status	Published - 2007 Jul

ASJC Scopus subject areas

Biochemistry, Genetics and Molecular Biology(all)

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10.1038/nprot.2007.199

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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.",

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Arabidopsis mesophyll protoplasts: A versatile cell system for transient gene expression analysis

Abstract

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