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Forced Air Microbiological Incubators

Company: VWR
Catalog#: 89511-430
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Assay for Assessing Mucin Binding to Bacteria and Bacterial Proteins
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Date:
2021-03-05
[Abstract]  

Legionella pneumophila, a Gram-negative bacterium and the causative agent of Legionnaires’ disease, exports over 300 effector proteins/virulence factors, through its type II (T2SS) and type IV secretion systems (T4SS). One such T2SS virulence factor, ChiA, not only functions as a chitinase, but also as a novel mucinase, which we believe aids ChiA-dependent virulence during lung infection. Previously published protocols manipulated wild-type L. pneumophila strain 130b and its chiA mutant to express plasmid-encoded GFP. Similarly, earlier studies demonstrated that wheat germ agglutinin (WGA) can be fluorescently labeled and can bind to mucins. In the current protocol, GFP-labeled bacteria were incubated with type II and type III porcine stomach mucins, which were then labeled with

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[摘要]  [摘要]嗜肺军团杆菌是革兰氏阴性细菌,是军团菌病的病原体,通过其II型(T2SS)和IV型分泌系统(T4SS)出口了300多种效应蛋白/毒力因子。一种这样的T2SS毒力因子ChiA不仅起几丁质酶的作用,而且还起新型粘蛋白酶的作用,我们认为它可以在肺部感染期间帮助ChiA依赖性毒力。以前发表的协议操纵野生型肺炎嗜血杆菌130b菌株及其chiA突变体,以表达质粒编码的GFP。同样,较早的研究表明,小麦胚芽凝集素(WGA)可以进行荧光标记,并可以与粘蛋白结合。 在当前方案中,将GFP标记的细菌与II型和III型猪胃粘蛋白孵育,然后用TexasRed标记的WGA进行标记,并通过流式细胞术进行分析,以测量在有或没有HLA的情况下细菌与粘蛋白的结合。内源性ChiA。另外,我们分析了纯化的ChiA与II型和III型猪胃粘蛋白的结合。该方案将细菌和直接蛋白结合到粘蛋白上,并且是第一个使用WGA和流式细胞术分析革兰氏阴性细菌与粘蛋白结合的方法。



图形摘要:

自动生成手机说明的屏幕截图

评估细菌和蛋白质与粘蛋白结合的策略


[背景技术]嗜肺军团菌(LPN ),革兰氏阴性细菌,是军团病,肺炎的严重形式的病原体。L ...

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