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

Sang Dong Yoo, Young Hee Cho, Jen Sheen

    Research output: Contribution to journalArticlepeer-review

    2919 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

    ASJC Scopus subject areas

    • Biochemistry, Genetics and Molecular Biology(all)

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