Abstract：
Eukaryote-interacting bacteria have developed along the evolution of an arsenal of tools to interact with potential hosts and to evade their defensive responses. Among these tools, the effector proteins are gaining a special importance due to the high diversity of molecular actions that they play in the host cell, with the final aim of taking the control over the cell. Bacteria inject these effectors into the cytosol of the host cells through distinct ways, as the type III secretion system. The study of the effectors\' molecular roles inside the host cell is challenging, due in part to the lack of traceability of such proteins once they are delivered by the bacteria. Here, we describe in depth a methodology that combines the increase of the bacterial effector concentration by protein expression systems with the use of heterologous hosts to facilitate the visualization of the subcellular targeting of the effector inside the host cell by fluorescence microscopy.
摘要：
与真核生物相互作用的细菌已经随着工具库的发展而发展，以与潜在宿主相互作用并逃避其防御反应。在这些工具中,由于它们在宿主细胞中发挥的分子作用的高度多样性，效应蛋白正变得特别重要，最终目的是控制细胞。细菌通过不同的方式将这些效应子注入宿主细胞的细胞质中，作为III型分泌系统。对宿主细胞内的效应分子作用的研究具有挑战性，部分原因是这些蛋白质一旦被细菌递送，就缺乏可追溯性。这里,我们深入描述了一种方法，该方法将通过蛋白质表达系统增加细菌效应物浓度与使用异源宿主相结合，以促进通过荧光显微镜观察宿主细胞内效应物的亚细胞靶向。
