Ubiquitin is a small protein commonly used as a signal molecule which upon attachment to the proteins affects their function and their fate in the cells. For example, it can be used as a degradation marker by the cell. Ubiquitin plays a significant role in regulation of numerous cellular processes. Therefore, monitoring of ubiquitin-dependent proteolysis can provide important information. Here, we describe construction of YFP-based proteasome substrates containing modified ubiquitin and the protocol for their transient expression in plant cells for functional analysis of the ubiquitin/proteasome system. To facilitate further subcloning all plasmids generated by us are based on the Gateway ® Cloning Technology and are compatible with the Gateway ® destination vectors.
CITATION STYLE
Zientara-Rytter, K., & Sirko, A. (2016). Fluorescent reporters for ubiquitin-dependent proteolysis in plants. In Methods in Molecular Biology (Vol. 1450, pp. 45–54). Humana Press Inc. https://doi.org/10.1007/978-1-4939-3759-2_5
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