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李斯特菌生物膜形成过程中细胞外 DNA 的作用。

Role of extracellular DNA during biofilm formation by Listeria monocytogenes.

机构信息

Food Microbiology, Department of Food Science, Faculty of Life Science, Copenhagen University, Rolighedsvej 30, DK-1958 Frederiksberg C, Denmark.

出版信息

Appl Environ Microbiol. 2010 Apr;76(7):2271-9. doi: 10.1128/AEM.02361-09. Epub 2010 Feb 5.

Abstract

Listeria monocytogenes is a food-borne pathogen that is capable of living in harsh environments. It is believed to do this by forming biofilms, which are surface-associated multicellular structures encased in a self-produced matrix. In this paper we show that in L. monocytogenes extracellular DNA (eDNA) may be the only central component of the biofilm matrix and that it is necessary for both initial attachment and early biofilm formation for 41 L. monocytogenes strains that were tested. DNase I treatment resulted in dispersal of biofilms, not only in microtiter tray assays but also in flow cell biofilm assays. However, it was also demonstrated that in a culture without eDNA, neither Listeria genomic DNA nor salmon sperm DNA by itself could restore the capacity to adhere. A search for additional necessary components revealed that peptidoglycan (PG), specifically N-acetylglucosamine (NAG), interacted with the DNA in a manner which restored adhesion. If a short DNA fragment (less than approximately 500 bp long) was added to an eDNA-free culture prior to addition of genomic or salmon sperm DNA, adhesion was prevented, indicating that high-molecular-weight DNA is required for adhesion and that the number of attachment sites on the cell surface can be saturated.

摘要

李斯特菌是一种食源性病原体,能够在恶劣的环境中生存。据信,它通过形成生物膜来实现这一点,生物膜是一种表面相关的多细胞结构,被自身产生的基质包裹。在本文中,我们表明,在李斯特菌中,细胞外 DNA(eDNA)可能是生物膜基质的唯一核心成分,并且对于 41 株李斯特菌的初始附着和早期生物膜形成都是必需的。DNase I 处理导致生物膜分散,不仅在微量滴定板测定中,而且在流动池生物膜测定中也是如此。然而,也证明了在没有 eDNA 的培养物中,李斯特菌基因组 DNA 或鲑鱼精子 DNA 本身都不能恢复粘附能力。对其他必需成分的搜索表明,肽聚糖(PG),特别是 N-乙酰葡萄糖胺(NAG),以一种恢复粘附的方式与 DNA 相互作用。如果在添加基因组或鲑鱼精子 DNA 之前,将一段短的 DNA 片段(小于约 500bp 长)添加到无 eDNA 的培养物中,粘附就会被阻止,这表明粘附需要高分子量的 DNA,并且细胞表面上的附着位点的数量可以被饱和。

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