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新型广谱 T 偶数噬菌体 S16 的长尾纤维特异性识别沙门氏菌 OmpC。

Long tail fibres of the novel broad-host-range T-even bacteriophage S16 specifically recognize Salmonella OmpC.

机构信息

Institute of Food, Nutrition and Health, ETH Zürich, Schmelzbergstrasse 7, 8092 Zürich, Switzerland.

出版信息

Mol Microbiol. 2013 Feb;87(4):818-34. doi: 10.1111/mmi.12134. Epub 2013 Jan 15.

Abstract

We report isolation and characterization of the novel T4-like Salmonella bacteriophage vB_SenM-S16. S16 features a T-even morphology and a highly modified 160 kbp dsDNA genome with 36.9 mol % G+C, containing 269 putative coding sequences and three tRNA genes. S16 is a virulent phage, and exhibits a maximally broad host range within the genus Salmonella, but does not infect other bacteria. Synthesis of functional S16 full-length long tail fibre (LTF) in Escherichia coli was possible by coexpression of gp37 and gp38. Surface plasmon resonance analysis revealed nanomolar equilibrium affinity of the LTF to its receptor on Salmonella cells. We show that OmpC serves as primary binding ligand, and that S16 adsorption can be transferred to E. coli by substitution of ompC with the Salmonella homologue. S16 also infects 'rough' Salmonella strains which are defective in lipopolysaccharide synthesis and/or its carbohydrate substitution, indicating that this interaction does not require an intact LPS structure. Altogether, its virulent nature, broad host range and apparent lack of host DNA transduction render S16 highly suitable for biocontrol of Salmonella in foods and animal production. The S16 LTF represents a highly specific affinity reagent useful for cell decoration and labelling, as well as bacterial immobilization and separation.

摘要

我们报告了新型 T4 样沙门氏菌噬菌体 vB_SenM-S16 的分离和特性。S16 具有 T 偶数形态和高度修饰的 160kbp dsDNA 基因组,GC 含量为 36.9 mol%,包含 269 个推定编码序列和三个 tRNA 基因。S16 是一种毒性噬菌体,在沙门氏菌属内表现出最大的广谱宿主范围,但不感染其他细菌。通过共表达 gp37 和 gp38,在大肠杆菌中合成功能性 S16 全长长尾纤维(LTF)是可能的。表面等离子体共振分析显示 LTF 与沙门氏菌细胞上受体的纳米级平衡亲和力。我们表明 OmpC 作为主要结合配体,并且通过用沙门氏菌同源物替代 ompC,可以将 S16 的吸附转移到大肠杆菌上。S16 还感染脂多糖合成和/或其碳水化合物取代缺陷的“粗糙”沙门氏菌菌株,表明这种相互作用不需要完整的 LPS 结构。总的来说,其毒性、广谱宿主范围和明显缺乏宿主 DNA 转导使 S16 非常适合在食品和动物生产中用于沙门氏菌的生物控制。S16 LTF 是一种高度特异性的亲和试剂,可用于细胞装饰和标记,以及细菌固定和分离。

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