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Author:
Date:
2014-10-20
[Abstract] Many gram-negative bacterial pathogens, including Shigella flexneri, are able to translocate bacterial proteins, dubbed effectors, across the host cell plasma membrane into the host cell cytosol using a syringe-like structure, the type three secretion apparatus (T3SA). While some bacteria use their T3SA to modulate their phagosomal environment (Salmonella spp.), establish pedestal structure to form microcolonies on the plasma membrane (Enteropathogenic Escherichi coli) or lyse their entry vacuole (Shigella spp.), they all have in common a tightly regulated activity of their T3SA. However, the tracking of the activity of the T3SA in infected cells and tissue has been difficult to perform. Using the property of MxiE-dependent promoters that are ...
[摘要] 包括灵芝氏菌在内的许多革兰氏阴性细菌病原体能够使用注射器样结构将类型三分泌物穿过宿主细胞质膜转运细菌蛋白质（配体效应子）到宿主细胞胞质溶胶中装置（T3SA）。虽然一些细菌使用它们的T3SA调节它们的吞噬体环境（沙门氏菌 spp 。），建立基质结构以在质膜上形成微集落（Enteropathogenic < em="">）或溶解它们的进入泡（志贺氏菌属。），它们都具有共同的严格调节的他们的T3SA的活性。然而，T3SA在感染的细胞和组织中的活性的跟踪一直难以进行。使用在T3SA是活性时上调的MxiE依赖性启动子的性质，我们最近设计了基于转录的分泌活性报道分子（TSAR），其允许以下的S的活性。使用快速成熟的GFP内在荧光，在组织培养细胞中实时地和体内实时分析灵敏度。在这里我们描述TSAR的设计及其应用于固定和活体样品的显微镜和流式细胞仪在结肠上皮细胞模型使用TC7组织培养细胞。
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