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Updated: Oct 1, 2025
In Vitro SUMOylation Assay to Study SUMO E3 Ligase Activity
Published on: January 29, 2018
An optimized protocol to assess SUMOylation in the plant Capsella rubella using two-component DEX-inducible
Yang Dong1,2, Zhi-Cheng Hu1,2, Lars Østergaard3
1State Key Laboratory of Systematic and Evolutionary Botany, Institute of Botany, Chinese Academy of Sciences, 20 Nanxincun, Xiangshan, Beijing 100093, China.
Abstract:
Here, we present an efficient protocol to test the SUMOylation of a target protein in the plant Capsella rubella based on overexpression of dexamethasone (DEX)-inducible tagged proteins. We describe the construction of two-component, FLAG-tagged DEX-inducible plasmids. We then detail the transformation of Capsella, followed by DEX treatment and SUMOylation assays. This protocol can be widely applied to proteins with expression restricted to specific cells and tissues using native promoters as well as proteins whose overexpression leads to embryo lethality. For complete details on the use and execution of this profile, please refer to Dong et al. (2020).
