Madec Edwige, Lacroix Jean-Marie, Bontemps-Gallo Sébastien
University Lille, CNRS, UMR 8576-UGSF-Unité de Glycobiologie Structurale et Fonctionnelle, Lille, France.
University Lille, CNRS, UMR 8576-UGSF-Unité de Glycobiologie Structurale et Fonctionnelle, Lille, France.
STAR Protoc. 2025 Jul 31;6(3):103999. doi: 10.1016/j.xpro.2025.103999.
Quantifying the phosphorylation levels of proteins involved in bacterial signaling pathways is essential for understanding their role in stress adaptation and infection processes. Here, we present a protocol for the semi-quantitative assessment of the phosphorylation level of the transcriptional regulator CpxR during the infection process using Phos-tag gel. We describe steps for Dickeya dadantii infection of chicory leaves, followed by bacterial extraction, rapid acidic cell lysis, and subsequent protein analysis using Phos-tag SDS-PAGE and western blot. For complete details on the use and execution of this protocol, please refer to Bontemps-Gallo et al. and Cochard et al..
量化参与细菌信号通路的蛋白质的磷酸化水平对于理解它们在应激适应和感染过程中的作用至关重要。在此，我们提出了一种使用Phos-tag凝胶对感染过程中转录调节因子CpxR的磷酸化水平进行半定量评估的方案。我们描述了菊苣叶被达旦氏菌感染的步骤，随后是细菌提取、快速酸性细胞裂解，以及随后使用Phos-tag SDS-PAGE和蛋白质印迹进行蛋白质分析。有关本方案使用和执行的完整详细信息，请参考邦滕普斯-加洛等人以及科沙尔等人的研究。
