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化学和低温处理及适应对大肠杆菌O157:H7毒力因子基因和外膜蛋白反应的影响

Effects of chemical and low-temperature treatments and adaption on the responses of virulence factor genes and outer membrane proteins in Escherichia coli O157:H7.

作者信息

Huang Da-Hao, Wang Kun, Chiu Chihwei P, Pan Tzu-Ming, Tsai Tsung-Yu

机构信息

Department of Food Science, Fu Jen Catholic University, New Taipei City, Taiwan.

Division of Gastroenterology, Department of Internal Medicine, Cardinal Tien Hospital, New Taipei City, Taiwan.

出版信息

J Microbiol Immunol Infect. 2015 Dec;48(6):604-12. doi: 10.1016/j.jmii.2014.03.007. Epub 2014 May 22.

Abstract

BACKGROUND

In the years after the discovery of this pathogen, Escherichia coli O157:H7 has become increasingly prominent, and outbreaks have been reported in many areas.

METHODS

In the current study, we determined the expressions of virulent factor genes shiga-like toxin II (stxII) and E. coli attaching and effacing (eaeA), in strains that were isolated from cattle feces and a clinical case in Taiwan. In addition, the effects of chemical and low-temperature stresses and adaptions on the expressions of virulent factor genes were investigated. Furthermore, the outer membrane proteins of acid-adapted E. coli O157:H7 TWC01 (TWC01) was separated using two-dimensional electrophoresis, and proteins were identified using mass spectrometry in order to illustrate the changes in protein expression after adaption.

RESULTS

Expressions of stxII and eaeA in the TWC01 isolated from a clinical case were higher than those in two strains isolated from cattle feces, and both organic and inorganic acid stresses and adaptions enhance the expression of genes encoding virulent factors in strains. In addition, the outer membrane proteins of TWC01 were regulated under hydrochloric acid adaption, indicating induction of acid tolerance and enhancement adhesion in TWC01. Lactic acid treatment of TWC01 resulted in downregulation of channel protein and adherence-related protein expressions.

CONCLUSION

The results of this study are helpful in understanding the resistance of locally isolated TWC01 to chemical and low-temperature stresses, and improving the control of this pathogen.

摘要

背景

在发现这种病原体后的数年里,大肠埃希菌O157:H7日益突出,许多地区都报告了疫情暴发。

方法

在本研究中,我们测定了从台湾牛粪便和一个临床病例中分离出的菌株中,毒力因子基因志贺样毒素II(stxII)和大肠埃希菌紧密黏附素(eaeA)的表达情况。此外,还研究了化学和低温应激及适应对毒力因子基因表达的影响。此外,利用二维电泳分离酸适应的大肠埃希菌O157:H7 TWC01(TWC01)的外膜蛋白,并通过质谱鉴定蛋白质,以阐明适应后蛋白质表达的变化。

结果

从临床病例中分离出的TWC01中stxII和eaeA的表达高于从牛粪便中分离出的两株菌株,有机酸和无机酸应激及适应均增强了菌株中毒力因子编码基因的表达。此外,TWC01的外膜蛋白在盐酸适应下受到调控,表明TWC01诱导了耐酸性并增强了黏附性。用乳酸处理TWC01导致通道蛋白和黏附相关蛋白表达下调。

结论

本研究结果有助于了解本地分离的TWC01对化学和低温应激的抗性,并改善对该病原体的控制。

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