title: Transient expression assays in tobacco protoplasts.
abstract: The sequence information generated through genome and transcriptome analysis from plant tissues has reached unprecedented sizes. Sequence homology-based annotations may provide hints for the possible function and roles of particular plant genes, but the functional annotation remains nonexistent or incomplete for many of them. To discover gene functions, transient expression assays are a valuable tool because they can be done more rapidly and at a higher scale than generating stably transformed tissues. Here, we describe a transient expression assay in protoplasts derived from suspension cells of tobacco (Nicotiana tabacum) for the study of the transactivation capacities of transcription factors. To enhance throughput and reproducibility, this method can be automated, allowing medium-throughput screening of interactions between large compendia of potential transcription factors and gene promoters.
authors.affiliation: Department of Plant Systems Biology, VIB, Gent, Belgium.
authors.forename: Robin
authors.initials: R
authors.lastname: Vanden Bossche
authors.forename: Brecht
authors.initials: B
authors.lastname: Demedts
authors.forename: Rudy
authors.initials: R
authors.lastname: Vanderhaeghen
authors.forename: Alain
authors.initials: A
authors.lastname: Goossens
mesh_terms: D018929:Cell Culture Techniques; D015870:Gene Expression; D017930:Genes, Reporter; D049409:Luciferases, Firefly / Q000096:biosynthesis / Q000235:genetics; D010940:Plant Proteins / Q000096:biosynthesis* / Q000235:genetics; D010957:Plasmids / Q000235:genetics; D011401:Promoter Regions, Genetic; D011523:Protoplasts / Q000378:metabolism*; D014026:Nicotiana / Q000166:cytology / Q000378:metabolism*; D014157:Transcription Factors / Q000096:biosynthesis* / Q000235:genetics; D014162:Transfection
chemical_list: D010940:Plant Proteins; D014157:Transcription Factors; D049409:Luciferases, Firefly