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Optimized Fluorescence Complementation Platform for Visualizing Salmonella Effector Proteins Reveals Distinctly Different Intracellular Niches in Different Cell Types.
ACS Infectious Diseases ( IF 5.3 ) Pub Date : 2017-06-09 00:00:00 , DOI: 10.1021/acsinfecdis.7b00052
Alexandra M Young 1 , Michael Minson 1 , Sarah E McQuate 1 , Amy E Palmer 1
Affiliation  

The bacterial pathogen Salmonella uses sophisticated type III secretion systems (T3SS) to translocate and deliver bacterial effector proteins into host cells to establish infection. Monitoring these important virulence determinants in the context of live infections is a key step in defining the dynamic interface between the host and pathogen. Here, we provide a modular labeling platform based on fluorescence complementation with split-GFP that permits facile tagging of new Salmonella effector proteins. We demonstrate enhancement of split-GFP complementation signals by manipulating the promoter or by multimerizing the fluorescent tag and visualize three effector proteins, SseF, SseG, and SlrP, that have never before been visualized over time during infection of live cells. Using this platform, we developed a methodology for visualizing effector proteins in primary macrophage cells for the first time and reveal distinct differences in the effector-defined intracellular niche between primary macrophage and commonly used HeLa and RAW cell lines.

中文翻译:

用于可视化沙门氏菌效应蛋白的优化荧光互补平台揭示了不同细胞类型中不同的细胞内壁ches。

细菌病原体沙门氏菌使用复杂的III型分泌系统(T3SS)来转运细菌效应蛋白并将其传递到宿主细胞中以建立感染。在活感染的情况下监视这些重要的毒力决定因素是定义宿主与病原体之间动态接口的关键步骤。在这里,我们提供了一个基于带有互补GFP的荧光互补的模块化标记平台,该平台可以方便地标记新的沙门氏菌。效应蛋白。我们通过操纵启动子或使荧光标记多聚化,并显示三种效应蛋白,SseF,SseG和SrrP,展示了分裂的GFP互补信号的增强,这三种蛋白在活细胞感染过程中不会随着时间的流逝而显现。使用该平台,我们首次开发了一种可视化初级巨噬细胞中效应蛋白的方法,并揭示了初级巨噬细胞与常用的HeLa和RAW细胞系之间的效应子定义的细胞内生态位的明显差异。
更新日期:2017-06-09
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