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对大肠杆菌 O157:H7 阪崎菌株在低温和水分活度胁迫的稳态条件下生理反应的转录组和蛋白质组综合分析。

Integrated transcriptomic and proteomic analysis of the physiological response of Escherichia coli O157:H7 Sakai to steady-state conditions of cold and water activity stress.

机构信息

Food Safety Centre, Tasmanian Institute of Agricultural Research, School of Agricultural Science, University of Tasmania, Private Bag 54, Hobart TAS 7001, Australia.

CSIRO Food and Nutritional Sciences, PO Box 52, North Ryde NSW 1670, Australia.

出版信息

Mol Cell Proteomics. 2012 Jan;11(1):M111.009019. doi: 10.1074/mcp.M111.009019. Epub 2011 Oct 18.

Abstract

An integrated transcriptomic and proteomic analysis was undertaken to determine the physiological response of Escherichia coli O157:H7 Sakai to steady-state conditions relevant to low temperature and water activity conditions experienced during meat carcass chilling in cold air. The response of E. coli during exponential growth at 25 °C a(w) 0.985, 14 °C a(w) 0.985, 25 °C a(w) 0.967, and 14 °C a(w) 0.967 was compared with that of a reference culture (35 °C a(w) 0.993). Gene and protein expression profiles of E. coli were more strongly affected by low water activity (a(w) 0.967) than by low temperature (14 °C). Predefined group enrichment analysis revealed that a universal response of E. coli to all test conditions included activation of the master stress response regulator RpoS and the Rcs phosphorelay system involved in the biosynthesis of the exopolysaccharide colanic acid, as well as down-regulation of elements involved in chemotaxis and motility. However, colanic acid-deficient mutants were shown to achieve comparable growth rates to their wild-type parents under all conditions, indicating that colanic acid is not required for growth. In contrast to the transcriptomic data, the proteomic data revealed that several processes involved in protein synthesis were down-regulated in overall expression at 14 °C a(w) 0.985, 25 °C a(w) 0.967, and 14 °C a(w) 0.967. This result suggests that during growth under these conditions, E. coli, although able to transcribe the required mRNA, may lack the cellular resources required for translation. Elucidating the global adaptive response of E. coli O157:H7 during exposure to chilling and water activity stress has provided a baseline of knowledge of the physiology of this pathogen.

摘要

进行了综合转录组学和蛋白质组学分析,以确定大肠杆菌 O157:H7 Sakai 对与低温和空气冷却肉胴体过程中经历的低水活度相关的稳定状态条件的生理反应。在 25°C(a(w)0.985)、14°C(a(w)0.985)、25°C(a(w)0.967)和 14°C(a(w)0.967)指数生长期的大肠杆菌的反应与参考培养物(35°C(a(w)0.993))进行了比较。与低温(14°C)相比,低水活度(a(w)0.967)对大肠杆菌的基因和蛋白质表达谱有更强的影响。预定义组富集分析表明,大肠杆菌对所有测试条件的普遍反应包括主应激反应调节剂 RpoS 的激活和涉及外多糖柯聚糖生物合成的 Rcs 磷酸传递系统,以及参与趋化性和运动性的元素下调。然而,柯聚糖缺陷突变体在所有条件下都显示出与野生型亲本相当的生长速率,表明柯聚糖不是生长所必需的。与转录组数据相反,蛋白质组数据显示,在 14°C(a(w)0.985)、25°C(a(w)0.967)和 14°C(a(w)0.967)时,几个参与蛋白质合成的过程在总体表达中被下调。这一结果表明,在这些条件下生长时,大肠杆菌虽然能够转录所需的 mRNA,但可能缺乏翻译所需的细胞资源。阐明大肠杆菌 O157:H7 在接触冷却和水活度应激时的全球适应反应为了解该病原体的生理学提供了一个基础。

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