title: Arabidopsis mesophyll protoplasts: a versatile cell system for transient gene expression analysis.
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.
authors.affiliation: Department of Molecular Biology, Massachusetts General Hospital, Boston, Massachusetts, USA. yoo@molbio.mgh.harvard.edu
authors.forename: Sang-Dong
authors.initials: SD
authors.lastname: Yoo
authors.forename: Young-Hee
authors.initials: YH
authors.lastname: Cho
authors.forename: Jen
authors.initials: J
authors.lastname: Sheen
mesh_terms: D017360:Arabidopsis / Q000166:cytology* / Q000235:genetics* / Q000378:metabolism; D029681:Arabidopsis Proteins / Q000235:genetics / Q000378:metabolism; D046508:Culture Techniques; D018506:Gene Expression Regulation, Plant; D011523:Protoplasts / Q000378:metabolism*; D015398:Signal Transduction; D014162:Transfection
chemical_list: D029681:Arabidopsis Proteins