Cargando…
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...
Autores principales: | , , |
---|---|
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 |
_version_ | 1784660513900199936 |
---|---|
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). |
format | Online Article Text |
id | pubmed-8885766 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Elsevier |
record_format | MEDLINE/PubMed |
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 |
work_keys_str_mv | AT dongyang anoptimizedprotocoltoassesssumoylationintheplantcapsellarubellausingtwocomponentdexinducibletransformants AT huzhicheng anoptimizedprotocoltoassesssumoylationintheplantcapsellarubellausingtwocomponentdexinducibletransformants AT østergaardlars anoptimizedprotocoltoassesssumoylationintheplantcapsellarubellausingtwocomponentdexinducibletransformants AT dongyang optimizedprotocoltoassesssumoylationintheplantcapsellarubellausingtwocomponentdexinducibletransformants AT huzhicheng optimizedprotocoltoassesssumoylationintheplantcapsellarubellausingtwocomponentdexinducibletransformants AT østergaardlars optimizedprotocoltoassesssumoylationintheplantcapsellarubellausingtwocomponentdexinducibletransformants |