title: Analysis of a transcription factor using transient assay in Arabidopsis protoplasts.
abstract: Regulation of gene expression by transcription factors is a fundamental mechanism in essentially all aspects of cellular processes. Transient expression assay of a reporter plasmid containing a reporter gene driven by a promoter of interest and an effector plasmid expressing a transcription factor has been a powerful tool for analyzing transcription factors. Here we present a protocol for polyethylene glycol (PEG)-mediated transformation of Arabidopsis protoplasts. It details preparation of protoplasts from Arabidopsis suspension cultured cells or leaves of soil-grown Arabidopsis plants and subsequent PEG-mediated transformation with reporter and effector plasmids. This protocol can be completed within 24 h from protoplast preparation to reporter assay. As an example, analysis of the membrane-bound transcription factor AtbZIP60 and its target BiP3 promoter is shown.
authors: Huck Institutes of the Life Sciences, Pennsylvania State University, University Park, PA, USA. yuji.iwata01@gmail.com
authors: Yuji
authors: Y
authors: Iwata
authors: Mi-Hyun
authors: MH
authors: Lee
authors: Nozomu
authors: N
authors: Koizumi
mesh_terms: D017360:Arabidopsis / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism; D029681:Arabidopsis Proteins / Q000235:genetics; D050976:Basic-Leucine Zipper Transcription Factors / Q000235:genetics; D018506:Gene Expression Regulation, Plant; D018014:Gene Transfer Techniques; D017930:Genes, Reporter / Q000235:genetics; D010957:Plasmids / Q000235:genetics; D011092:Polyethylene Glycols / Q000494:pharmacology; D011401:Promoter Regions, Genetic; D011523:Protoplasts / Q000166:cytology / Q000378:metabolism*; D014157:Transcription Factors / Q000235:genetics* / Q000378:metabolism; D014162:Transfection
chemical_list: D029681:Arabidopsis Proteins; D050976:Basic-Leucine Zipper Transcription Factors; D014157:Transcription Factors; C533092:bZIP60 protein, Arabidopsis; D011092:Polyethylene Glycols