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

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 transformat...

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Detalles Bibliográficos
Autores principales: Dong, Yang, Hu, Zhi-Cheng, Østergaard, Lars
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8885766/
https://www.ncbi.nlm.nih.gov/pubmed/35243380
http://dx.doi.org/10.1016/j.xpro.2022.101197
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author Dong, Yang
Hu, Zhi-Cheng
Østergaard, Lars
author_facet Dong, Yang
Hu, Zhi-Cheng
Østergaard, Lars
author_sort Dong, Yang
collection PubMed
description 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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spelling pubmed-88857662022-03-02 An optimized protocol to assess SUMOylation in the plant Capsella rubella using two-component DEX-inducible transformants Dong, Yang Hu, Zhi-Cheng Østergaard, Lars STAR Protoc Protocol 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). Elsevier 2022-02-26 /pmc/articles/PMC8885766/ /pubmed/35243380 http://dx.doi.org/10.1016/j.xpro.2022.101197 Text en © 2022 The Author(s) https://creativecommons.org/licenses/by/4.0/This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Protocol
Dong, Yang
Hu, Zhi-Cheng
Østergaard, Lars
An optimized protocol to assess SUMOylation in the plant Capsella rubella using two-component DEX-inducible transformants
title An optimized protocol to assess SUMOylation in the plant Capsella rubella using two-component DEX-inducible transformants
title_full An optimized protocol to assess SUMOylation in the plant Capsella rubella using two-component DEX-inducible transformants
title_fullStr An optimized protocol to assess SUMOylation in the plant Capsella rubella using two-component DEX-inducible transformants
title_full_unstemmed An optimized protocol to assess SUMOylation in the plant Capsella rubella using two-component DEX-inducible transformants
title_short An optimized protocol to assess SUMOylation in the plant Capsella rubella using two-component DEX-inducible transformants
title_sort optimized protocol to assess sumoylation in the plant capsella rubella using two-component dex-inducible transformants
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8885766/
https://www.ncbi.nlm.nih.gov/pubmed/35243380
http://dx.doi.org/10.1016/j.xpro.2022.101197
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