An optimized protocol to assess SUMOylation in the plant Capsella rubella using two-component DEX-inducible transformants
Summary: 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 tr...
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| Format: | Article |
| Language: | English |
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Elsevier
2022-03-01
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| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166722000776 |
| _version_ | 1818774519631839232 |
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| author | Yang Dong Zhi-Cheng Hu Lars Østergaard |
| author_facet | Yang Dong Zhi-Cheng Hu Lars Østergaard |
| author_sort | Yang Dong |
| collection | DOAJ |
| description | Summary: 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). |
| first_indexed | 2024-12-18T10:42:26Z |
| format | Article |
| id | doaj.art-19c07c8b248d49ec80195ab77408b0f0 |
| institution | Directory Open Access Journal |
| issn | 2666-1667 |
| language | English |
| last_indexed | 2024-12-18T10:42:26Z |
| publishDate | 2022-03-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj.art-19c07c8b248d49ec80195ab77408b0f02022-12-21T21:10:36ZengElsevierSTAR Protocols2666-16672022-03-0131101197An optimized protocol to assess SUMOylation in the plant Capsella rubella using two-component DEX-inducible transformantsYang Dong0Zhi-Cheng Hu1Lars Østergaard2State Key Laboratory of Systematic and Evolutionary Botany, Institute of Botany, Chinese Academy of Sciences, 20 Nanxincun, Xiangshan, Beijing 100093, China; University of Chinese Academy of Sciences, Beijing 100049, China; Corresponding authorState Key Laboratory of Systematic and Evolutionary Botany, Institute of Botany, Chinese Academy of Sciences, 20 Nanxincun, Xiangshan, Beijing 100093, China; University of Chinese Academy of Sciences, Beijing 100049, ChinaCrop Genetics Department, John Innes Centre, NR4 7UH Norwich, UK; Corresponding authorSummary: 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).http://www.sciencedirect.com/science/article/pii/S2666166722000776Model OrganismsPlant sciencesMolecular BiologyProtein Biochemistry |
| spellingShingle | Yang Dong Zhi-Cheng Hu Lars Østergaard An optimized protocol to assess SUMOylation in the plant Capsella rubella using two-component DEX-inducible transformants STAR Protocols Model Organisms Plant sciences Molecular Biology Protein Biochemistry |
| 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 | Model Organisms Plant sciences Molecular Biology Protein Biochemistry |
| url | http://www.sciencedirect.com/science/article/pii/S2666166722000776 |
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