An optimized protocol to assess SUMOylation in the plant Capsella rubella using two-component DEX-inducible transformants.
An optimized protocol to assess SUMOylation in the plant Capsella rubella using two-component DEX-inducible transformants.
复制标题
一个优化的协议，以评估SUMO化在植物荠菜风疹使用双组分DEX诱导转化。
DOI：
10.1016/j.xpro.2022.101197
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发表时间：
2022-03-18
期刊：
影响因子：
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通讯作者：
Østergaard L
中科院分区：
文献类型：
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作者：
Dong Y;Hu ZC;Østergaard L
在这里，我们提出了一种有效的方案来测试一个目标蛋白的SUMOylation在风疹甘蓝基于地塞米松（DEX）诱导标记蛋白的过表达。我们描述了双组分，flag标记dex诱导质粒的构建。然后我们详细介绍了辣椒的转化，随后进行了DEX处理和SUMOylation分析。该方案可广泛应用于使用天然启动子表达仅限于特定细胞和组织的蛋白质，以及过度表达导致胚胎致死的蛋白质。有关使用和执行此配置文件的完整详细信息，请参阅。在此，我们提出了一种基于地塞米松（DEX）诱导标记蛋白过表达的高效方案，来测试植物风疹中目标蛋白的summoylation。我们描述了双组分，flag标记dex诱导质粒的构建。然后我们详细介绍了辣椒的转化，随后进行了DEX处理和SUMOylation分析。该方案可广泛应用于使用天然启动子表达仅限于特定细胞和组织的蛋白质，以及过度表达导致胚胎致死的蛋白质。
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. An easy-to-use pipeline for constructing FLAG-tagged DEX-inducible plasmids An efficient transformation protocol for Capsella rubella An optimized protocol to test SUMOylation of a protein of interest in plants 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.