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Analysis of a Transcription Factor Using Transient Assay in Arabidopsis Protoplasts

 Last Update: 2020-11-07
 Source: Internet
 Author: User

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polyethylene

methylation gene expression

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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.

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