发布: 2014年11月20日第4卷第22期 DOI: 10.21769/BioProtoc.1302 浏览次数: 11101
相关实验方案
通过制备连续聚丙烯酰胺凝胶电泳和凝胶酶谱分析法纯化来自梭状龋齿螺旋体的天然Dentilisin复合物及其功能分析
Pachiyappan Kamarajan [...] Yvonne L. Kapila
2024年04月05日 1040 阅读
Abstract
Shigella flexneri (S. flexneri) is a Gram-negative bacterium that causes gastroenteritis and shigellosis in humans. In order to establish and maintain an infection, S. flexneri utilises a type three secretion system (T3SS) to deliver virulence factors called effector proteins into the cytoplasm of host cells, facilitating e.g. uptake into the host cell and escape from the endosome. Secretion through the T3SS is tightly regulated and is usually triggered by host-cell contact, but can also be artificially stimulated in vitro. In this assay, the dye Congo red is used to induce T3SS-dependent secretion of S. flexneri (Parsot et al., 1995) and secreted proteins are concentrated from the culture supernatant by precipitation with trichloroacetic acid. The assay presented here can easily be adapted to the secretion analysis of other bacteria utilising a T3SS, such as Salmonella typhimurium, which constitutively secrete when grown at 37 °C (Collazo et al., 1995; Pegues et al., 1995), or pathogenic species of Yersinia, where secretion can be induced by calcium deprivation (Heesemann et al., 1986; Forsberg et al., 1987).
Keywords: Shigella flexneri (福氏志贺菌)Materials and Reagents
Equipment
Procedure
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© 2014 The Authors; exclusive licensee Bio-protocol LLC.
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分类
微生物学 > 微生物-宿主相互作用 > 细菌
微生物学 > 微生物生物化学 > 蛋白质
分子生物学 > 蛋白质 > 检测
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