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迟缓爱德华氏菌 mltA 过表达降低了对抗生素的耐药性,并增强了斑马鱼的致死率。

Overexpression of mltA in Edwardsiella tarda reduces resistance to antibiotics and enhances lethality in zebra fish.

机构信息

College of Marine Life Sciences, Ocean University of China, Qingdao, China.

出版信息

J Appl Microbiol. 2012 Jun;112(6):1075-85. doi: 10.1111/j.1365-2672.2012.05291.x. Epub 2012 Apr 20.

Abstract

AIMS

The aim of this study was to investigate the role of membrane-bound lytic murein transglycosylase A (MltA) in a bacterial fish pathogen Edwardsiella tarda.

METHODS AND RESULTS

An mltA in-frame deletion mutant (ΔmltA) and an mltA overexpression strain (mltA(+)) of Edw. tarda were constructed through double-crossover allelic exchange and by transformation of a low-copy plasmid carrying the intact mltA into the ΔmltA mutant, respectively. Either inactivation or overexpression of MltA in Edw. tarda resulted in elevated sensitivity to β-lactam antibiotics and lower viability in oligotrophic or high osmotic environment than wild-type strain. Autolysis induced by EDTA was reduced in ΔmltA strain, while mltA(+) strain was virtually flimsy, indicating that MltA is responsible for the lysis effect. Moreover, mltA(+) strain exhibited significant increases in lipopolysaccharide (LPS) biosynthesis and virulence to zebra fish compared with wild-type strain.

CONCLUSIONS

The results indicated that MltA plays essential roles in β-lactam antibiotics and environmental stresses resistance, autolysis, LPS biosynthesis and pathogenicity of Edw. tarda. This is the first report that MltA has a virulence-related function in Edw. tarda.

SIGNIFICANCE AND IMPACT OF THE STUDY

This study provided useful information for further studies on pathogenesis of Edw. tarda.

摘要

目的

本研究旨在探讨膜结合溶菌壁转糖基酶 A(MltA)在鱼类病原菌爱德华氏菌中的作用。

方法和结果

通过双交换基因同源重组构建了 Edw. tarda 的 mltA 缺失突变株(ΔmltA)和 mltA 过表达株(mltA(+)),并通过转化带有完整 mltA 的低拷贝质粒将 mltA(+)转入 ΔmltA 突变株中。Edw. tarda 中 MltA 的失活或过表达导致其对β-内酰胺类抗生素的敏感性增加,在贫营养或高渗环境中的生存能力降低。与野生型菌株相比,ΔmltA 菌株的 EDTA 诱导自溶减少,而 mltA(+)菌株几乎脆弱,表明 MltA 负责溶菌作用。此外,mltA(+)菌株的脂多糖(LPS)生物合成和对斑马鱼的毒力与野生型菌株相比显著增加。

结论

结果表明,MltA 在 Edw. tarda 对β-内酰胺类抗生素和环境胁迫的抗性、自溶、LPS 生物合成和致病性中起重要作用。这是首次报道 MltA 在 Edw. tarda 中具有与毒力相关的功能。

研究的意义和影响

本研究为进一步研究 Edw. tarda 的发病机制提供了有用的信息。

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