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空肠弯曲菌插入突变体文库的高通量测序显示 mapA 是鸡定植的适合度因素。

High-throughput sequencing of Campylobacter jejuni insertion mutant libraries reveals mapA as a fitness factor for chicken colonization.

机构信息

Department of Microbiology and Immunology, University of Michigan Medical School, Ann Arbor, Michigan, USA.

出版信息

J Bacteriol. 2014 Jun;196(11):1958-67. doi: 10.1128/JB.01395-13. Epub 2014 Mar 14.

Abstract

Campylobacter jejuni is a leading cause of gastrointestinal infections worldwide, due primarily to its ability to asymptomatically colonize the gastrointestinal tracts of agriculturally relevant animals, including chickens. Infection often occurs following consumption of meat that was contaminated by C. jejuni during harvest. Because of this, much interest lies in understanding the mechanisms that allow C. jejuni to colonize the chicken gastrointestinal tract. To address this, we generated a C. jejuni transposon mutant library that is amenable to insertion sequencing and introduced this mutant pool into day-of-hatch chicks. Following deep sequencing of C. jejuni mutants in the cecal outputs, several novel factors required for efficient colonization of the chicken gastrointestinal tract were identified, including the predicted outer membrane protein MapA. A mutant strain lacking mapA was constructed and found to be significantly reduced for chicken colonization in both competitive infections and monoinfections. Further, we found that mapA is required for in vitro competition with wild-type C. jejuni but is dispensable for growth in monoculture.

摘要

空肠弯曲菌是全球范围内引起胃肠道感染的主要原因,主要是因为它能够在农业相关动物(包括鸡)的胃肠道中无症状地定植。感染通常发生在食用了在收获过程中被空肠弯曲菌污染的肉之后。因此,人们对了解允许空肠弯曲菌定植鸡胃肠道的机制非常感兴趣。为了解决这个问题,我们生成了一个可进行插入测序的空肠弯曲菌转座子突变体文库,并将这个突变体库引入了刚出壳的小鸡中。在对盲肠排泄物中的空肠弯曲菌突变体进行深度测序后,鉴定出了几个在鸡胃肠道中有效定植所需的新的因素,包括预测的外膜蛋白 MapA。构建了一个缺失 mapA 的突变株,并发现该突变株在竞争感染和单感染中对鸡的定植能力显著降低。此外,我们发现 mapA 是与野生型空肠弯曲菌进行体外竞争所必需的,但在纯培养中是可有可无的。

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