Bacteriophage host range evolution through engineered enrichment bias, exploiting heterologous surface receptor expression.
Bacteriophage host range evolution through engineered enrichment bias, exploiting heterologous surface receptor expression.
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DOI：
10.1111/1462-2920.15188
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发表时间：
2020-08
影响因子：
5.1
通讯作者：
Ziyue Zeng;G. Salmond
中科院分区：
文献类型：
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作者：
Ziyue Zeng;G. Salmond
最初的噬菌体-宿主相互作用的研究已经在有限的噬菌体及其同源受体库上进行，例如噬菌体λ和大肠杆菌LamB（EcLamB）蛋白。除了噬菌体λ，对靶向EcLamB的其他噬菌体知之甚少。在这里，我们开发了一种简单的方法，用于以可预测的方式分离新的环境毒素，即分离靶向细菌特定受体的毒素，在这种情况下，是EcLamB蛋白。将编码EcLamB孔蛋白的质粒（pMUT 13）转移到三种不同的肠细菌属中。通过富集这些工程细菌，可以很容易地从环境中分离出许多针对EcLamB的噬菌体（ZZ噬菌体）。有趣的是，虽然在重组异源宿主中依赖于EcLamB，但这些新分离的ZZ质粒在感染E.杆菌此外，噬菌体宿主范围很容易扩展到三个不同的细菌属异源表达的EcLamB。与噬菌体λ不同，噬菌体λ是Siphoviridae家族的成员，基于透射电子显微镜和基因组序列，这些新分离的EcLamB依赖性噬菌体更常见于Myoviridae家族的成员。对这种方便、有效的噬菌体富集方法进行改进，可能有助于发现新的噬菌体。本文受版权保护。All rights reserved.
Research on the initial phage-host interaction has been conducted on a limited repertoire of phages and their cognate receptors, such as phage λ and the Escherichia coli LamB (EcLamB) protein. Apart from phage λ, little is known about other phages which target EcLamB. Here, we developed a simple method for isolating novel environmental phages in a predictable way, i.e. isolating phages which target a particular receptor(s) of a bacterium, in this case, the EcLamB protein. A plasmid (pMUT13) encoding the EcLamB porin was transferred into three different enterobacterial genera. By enrichment with these engineered bacteria, a number of phages (ZZ phages) that targeted EcLamB were easily isolated from the environment. Interestingly, although EcLamB-dependent in their recombinant heterologous hosts, these newly isolated ZZ phages also targeted OmpC as an alternative receptor when infecting E. coli. Moreover, the phage host range was readily extended within three different bacterial genera with heterologously expressed EcLamB. Unlike phage λ, which is a member of the Siphoviridae family, these newly isolated EcLamB-dependent phages were more commonly members of the Myoviridae family, based on transmission electron microscopy and genomic sequences. Modifications of this convenient and efficient phage enrichment method could be useful for discovery of novel phages. This article is protected by copyright. All rights reserved.