title: Isolation, culture, and transient transformation of plant protoplasts.
abstract: Transient gene expression in protoplasts, which has been used in several plant species, is an important and versatile tool for rapid functional gene analysis, protein subcellular localization, and biochemical manipulations. This unit describes transient gene expression by electroporation of DNA into protoplasts of Arabidopsis or tobacco suspension-cultured cells and by polyethylene glycol (PEG)-mediated DNA transformation into protoplasts derived from rice leaf sheaths. PEG-mediated DNA transformation for transient gene expression in rice protoplasts in suspension culture is also described as an alternative technique. Methods for collecting intracellular and secreted proteins are also provided.
authors.affiliation: School of Life Sciences, Centre for Cell and Developmental Biology and State Key Laboratory of Agrobiotechnology, The Chinese University of Hong Kong, Hong Kong, China.
authors.forename: Jinbo
authors.initials: J
authors.lastname: Shen
authors.forename: Jiaxin
authors.initials: J
authors.lastname: Fu
authors.forename: Jin
authors.initials: J
authors.lastname: Ma
authors.forename: Xiangfeng
authors.initials: X
authors.lastname: Wang
authors.forename: Caiji
authors.initials: C
authors.lastname: Gao
authors.forename: ChuXiong
authors.initials: C
authors.lastname: Zhuang
authors.forename: Jianmin
authors.initials: J
authors.lastname: Wan
authors.forename: Liwen
authors.initials: L
authors.lastname: Jiang
mesh_terms: D017360:Arabidopsis / Q000166:cytology / Q000235:genetics / Q000378:metabolism; D018929:Cell Culture Techniques / Q000379:methods*; D002469:Cell Separation / Q000379:methods*; D018014:Gene Transfer Techniques; D059828:Plant Cells / Q000378:metabolism*; D011523:Protoplasts / Q000166:cytology / Q000378:metabolism; D014026:Nicotiana / Q000166:cytology / Q000378:metabolism
keywords: Arabidopsis; PEG; electroporation; protoplast isolation; rice; tobacco; transient expression