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一种用鼠伤寒沙门氏菌感染秀丽隐杆线虫的实验方案。

A protocol to infect Caenorhabditis elegans with Salmonella typhimurium.

作者信息

Zhang Jiuli, Jia Kailiang

机构信息

Department of Biological Sciences, Florida Atlantic University.

Department of Biological Sciences, Florida Atlantic University;

出版信息

J Vis Exp. 2014 Jun 26(88):e51703. doi: 10.3791/51703.

Abstract

In the last decade, C. elegans has emerged as an invertebrate organism to study interactions between hosts and pathogens, including the host defense against gram-negative bacterium Salmonella typhimurium. Salmonella establishes persistent infection in the intestine of C. elegans and results in early death of infected animals. A number of immunity mechanisms have been identified in C. elegans to defend against Salmonella infections. Autophagy, an evolutionarily conserved lysosomal degradation pathway, has been shown to limit the Salmonella replication in C. elegans and in mammals. Here, a protocol is described to infect C. elegans with Salmonella typhimurium, in which the worms are exposed to Salmonella for a limited time, similar to Salmonella infection in humans. Salmonella infection significantly shortens the lifespan of C. elegans. Using the essential autophagy gene bec-1 as an example, we combined this infection method with C. elegans RNAi feeding approach and showed this protocol can be used to examine the function of C. elegans host genes in defense against Salmonella infection. Since C. elegans whole genome RNAi libraries are available, this protocol makes it possible to comprehensively screen for C. elegans genes that protect against Salmonella and other intestinal pathogens using genome-wide RNAi libraries.

摘要

在过去十年中,秀丽隐杆线虫已成为一种用于研究宿主与病原体相互作用的无脊椎动物,包括宿主对革兰氏阴性菌鼠伤寒沙门氏菌的防御。沙门氏菌在秀丽隐杆线虫的肠道中建立持续感染,并导致受感染动物过早死亡。秀丽隐杆线虫中已鉴定出多种免疫机制来抵御沙门氏菌感染。自噬是一种进化上保守的溶酶体降解途径,已被证明可限制沙门氏菌在秀丽隐杆线虫和哺乳动物中的复制。在此,描述了一种用鼠伤寒沙门氏菌感染秀丽隐杆线虫的方法,其中线虫在有限时间内接触沙门氏菌,类似于人类的沙门氏菌感染。沙门氏菌感染显著缩短了秀丽隐杆线虫的寿命。以必需的自噬基因bec-1为例,我们将这种感染方法与秀丽隐杆线虫RNAi喂食方法相结合,并表明该方法可用于检测秀丽隐杆线虫宿主基因在抵御沙门氏菌感染中的功能。由于有秀丽隐杆线虫全基因组RNAi文库,该方法使得使用全基因组RNAi文库全面筛选秀丽隐杆线虫中抵御沙门氏菌和其他肠道病原体的基因成为可能。

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