An optimized protocol to assess SUMOylation in the plant Capsella rubella using two-component DEX-inducible transformants

使用双组分 DEX 诱导转化体评估植物 Capsella rubella 中 SUMO 化修饰的优化方案

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作者:Yang Dong, Zhi-Cheng Hu, Lars Østergaard

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).

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