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基于羟自由基和超氧阴离子的氧化应激对对数期和静止期大肠杆菌 O157:H7 应激和毒力基因表达的影响。

Impact of hydroxyl- and superoxide anion-based oxidative stress on logarithmic and stationary phase Escherichia coli O157:H7 stress and virulence gene expression.

机构信息

Food, Nutrition and Health Program, Faculty of Land and Food Systems, University of British Columbia, 218-2205 East Mall, Vancouver, British Columbia, Canada.

出版信息

Food Microbiol. 2012 Feb;29(1):141-7. doi: 10.1016/j.fm.2011.09.014. Epub 2011 Sep 29.

Abstract

Targeted microarray analysis was used to examine growth phase-related transcriptional responses of Escherichia coli O157:H7 stress and virulence genes. Logarithmic and stationary phase cells were exposed to hydrogen peroxide (H(2)O(2); 1 and 2.5 mM) and paraquat (PQ; 350 μM) for 40 min. For both oxidants, more genes were significantly induced in logarithmic cells compared to cells in stationary phase. In total, 14 genes at 1 mM H(2)O(2) and 15 genes at 2.5 mM H(2)O(2) were significantly induced in logarithmic cells, with 11 affected genes common between treatments, including stx2a and stx2b. At 2.5 mM H(2)O(2), additional virulence genes were induced, including stx1a, stx1b, lpfA, a putative enterotoxin, and ler. Stx2 ELISA showed transcriptional induction of stx2a and stx2b correlated with significantly increased Stx2 in cultures exposed to 1 and 2.5 mM H(2)O(2). Logarithmic cells significantly induced 15 genes following PQ exposure, 12 of which were induced in H(2)O(2)-treated cells, including several virulence genes (stx1a, stx2a, and stx2b). Exposure of stationary phase cells to H(2)O(2) and PQ had little effect on the queried transcriptome, demonstrating the importance of considering this inherently stress resistant state when evaluating food safety interventions.

摘要

采用靶向微阵列分析技术,研究了大肠杆菌 O157:H7 应激和毒力基因的生长阶段相关转录反应。对数期和稳定期细胞分别暴露于过氧化氢(H2O2;1 和 2.5 mM)和百草枯(PQ;350 μM)40 分钟。对于这两种氧化剂,对数期细胞中诱导的基因明显多于稳定期细胞。在 1 mM H2O2 下,共有 14 个基因和在 2.5 mM H2O2 下有 15 个基因在对数期细胞中明显被诱导,其中 11 个受影响的基因在两种处理中是共同的,包括 stx2a 和 stx2b。在 2.5 mM H2O2 下,还诱导了其他毒力基因,包括 stx1a、stx1b、lpfA、一种假定的肠毒素和 ler。stx2 ELISA 显示,stx2a 和 stx2b 的转录诱导与暴露于 1 和 2.5 mM H2O2 的培养物中 Stx2 的显著增加相关。对数期细胞在 PQ 暴露后显著诱导了 15 个基因,其中 12 个基因在 H2O2 处理的细胞中被诱导,包括几个毒力基因(stx1a、stx2a 和 stx2b)。稳定期细胞暴露于 H2O2 和 PQ 对所查询的转录组几乎没有影响,这表明在评估食品安全干预措施时,考虑到这种固有的抗应激状态非常重要。

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