CEA, CNRS, Aix Marseille University Lab of Microbial Ecology of the Rhizosphere (LEMiRE), UMR7265 BIAM, Saint-Paul-lez-Durance, France.
Methods Mol Biol. 2024;2741:11-24. doi: 10.1007/978-1-0716-3565-0_2.
Outer membrane vesicles (OMVs), produced by Gram negative-bacteria and sRNAs, are key players in cell-to-cell communication and interactions of bacteria with the environment. OMVs act as information carriers and encapsulate various molecules such as proteins, lipids, metabolites, and RNAs. OMVs and sRNAs play a broad range of functions from pathogenesis to stress resistance, to biofilm formation and both mediate interkingdom signaling. Various studies indicate that there is a mechanism of intercellular communication mediated by OMV-derived bacterial RNAs that is conserved among certain bacterial species. Here we describe methods for the extraction and purification of vesicles produced by Gram-negative bacteria, such as Pseudomonas brassicacearum and Escherichia coli, and address methods for the extraction of OMVs-derived sRNA and techniques for the analysis of sRNAs.
外膜囊泡（OMVs）是革兰氏阴性菌产生的物质，也是小 RNA（sRNAs）的重要组成部分，它们在细胞间的通讯以及细菌与环境的相互作用中扮演着关键角色。OMVs 作为信息载体，可包裹多种分子，如蛋白质、脂质、代谢物和 RNA。OMVs 和 sRNAs 具有广泛的功能，从发病机制到抗应激能力，再到生物膜的形成，它们都能介导种间信号传递。多项研究表明，某些细菌种间存在一种由 OMV 衍生的细菌 RNA 介导的细胞间通讯机制。在这里，我们描述了从革兰氏阴性菌（如铜绿假单胞菌和大肠杆菌）中提取和纯化囊泡的方法，并介绍了提取 OMVs 衍生的 sRNA 的方法和 sRNA 分析技术。
