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转录组测序揭示了肠炎沙门氏菌血清型在抗菌蛋清中的生存机制。

Transcriptional Sequencing Uncovers Survival Mechanisms of Salmonella enterica Serovar Enteritidis in Antibacterial Egg White.

机构信息

MOST-USDA Joint Research Center for Food Safety, School of Agriculture & Biology, and State Key Lab of Microbial Metabolism, Shanghai Jiao Tong University, Shanghai, China.

Department of Bioinformatics and Biostatistics, School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai, China.

出版信息

mSphere. 2019 Feb 13;4(1):e00700-18. doi: 10.1128/mSphere.00700-18.

Abstract

The survival mechanism of serovar Enteritidis in antibacterial egg white is not fully understood. In our lab, an egg white-resistant strain, Enteritidis SJTUF 10978, was identified. Cell envelope damage and osmotic stress response (separation of cell wall and inner membrane as well as cytoplasmic shrinkage) of this strain surviving in egg white were identified through microscopic observation. RNA-Seq analysis of the transcriptome of survival in egg white showed that a considerable number of genes involved in DNA damage repair, alkaline pH adaptation, osmotic stress adaptation, envelope damage repair, pathogenicity island 2 (SPI-2), iron absorption, and biotin synthesis were significantly upregulated (fold change ≥ 2) in egg white, indicating that these pathways or genes might be critical for bacterial survival. RNA-Seq results were confirmed by qRT-PCR, and the survival analysis of six gene deletion mutants confirmed their importance in the survival of bacteria in egg white. The importance of alkaline pH adaptation and envelope damage repair for to survive in egg white were further confirmed by analysis of , , , and deletion mutants. According to the RNA-Seq results, we propose that alkaline pH adaptation might be the cause of bacterial osmotic stress phenotype and that the synergistic effect between alkaline pH and other inhibitory factors can enhance the bacteriostatic effect of egg white. Moreover, and were recognized as the central regulators that coordinate bacterial metabolism to adapt to envelope damage and alkaline pH. serovar Enteritidis is a major foodborne pathogen that causes salmonellosis mainly through contaminated chicken eggs or egg products and has been a worldwide public health threat since 1980. Frequent outbreaks of this serotype through eggs correlate significantly with its exceptional survival ability in the antibacterial egg white. Research on the survival mechanism of Enteritidis in egg white will help to further understand the complex and highly effective antibacterial mechanisms of egg white and lay the foundation for the development of safe and effective vaccines to prevent egg contamination by this serotype. Key pathways and genes that were previously overlooked under bactericidal conditions were characterized as being induced in egg white, and synergistic effects between different antimicrobial factors appear to exist according to the gene expression changes. Our work provides new insights into the survival mechanism of Enteritidis in egg white.

摘要

肠炎沙门氏菌在抗菌蛋清中的生存机制尚未完全阐明。在我们的实验室中,鉴定出了一种蛋清抗性菌株肠炎沙门氏菌 SJTUF 10978。通过显微镜观察,鉴定了该菌株在蛋清中存活时的细胞包膜损伤和渗透胁迫反应(细胞壁和内膜分离以及细胞质收缩)。生存于蛋清中的转录组 RNA-Seq 分析表明,大量参与 DNA 损伤修复、碱性 pH 适应、渗透胁迫适应、包膜损伤修复、致病岛 2(SPI-2)、铁吸收和生物素合成的基因显著上调(倍数变化≥2)在蛋清中,表明这些途径或基因可能对细菌的生存至关重要。RNA-Seq 结果通过 qRT-PCR 得到了验证,六个基因缺失突变体的生存分析证实了它们在细菌在蛋清中生存的重要性。通过分析碱性 pH 适应和包膜损伤修复对细菌在蛋清中生存的重要性,进一步证实了碱性 pH 适应和 envelope damage repair 的重要性。根据 RNA-Seq 结果,我们提出碱性 pH 适应可能是细菌渗透胁迫表型的原因,碱性 pH 与其他抑制因素的协同作用可以增强蛋清的抑菌作用。此外,和被认为是协调细菌代谢以适应包膜损伤和碱性 pH 的中央调节剂。肠炎沙门氏菌是一种主要的食源性病原体,主要通过受污染的鸡蛋或蛋制品引起沙门氏菌病,自 1980 年以来一直是全球公共卫生威胁。该血清型通过鸡蛋频繁爆发与它在抗菌蛋清中的异常生存能力密切相关。研究肠炎沙门氏菌在蛋清中的生存机制将有助于进一步了解蛋清复杂而高效的抗菌机制,并为开发预防该血清型鸡蛋污染的安全有效的疫苗奠定基础。以前在杀菌条件下被忽视的关键途径和基因在蛋清中被描述为被诱导,根据基因表达的变化,不同抗菌因子之间似乎存在协同作用。我们的工作为肠炎沙门氏菌在蛋清中的生存机制提供了新的见解。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c193/6374596/615cdeb6f3d2/mSphere.00700-18-f0001.jpg

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