College of Veterinary Medicine, Yangzhou Universitygrid.268415.c, Yangzhou, China.
Jiangsu Co-Innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou, China.
Microbiol Spectr. 2022 Oct 26;10(5):e0141022. doi: 10.1128/spectrum.01410-22. Epub 2022 Sep 7.
The P1-like phage plasmid (PP) has been widely used as a molecular biology tool, but its role as an active accessory cargo element is not fully understood. In this study, we provide insights into the structural features and gene content similarities of 77 P1-like PPs in the RefSeq database. We also describe a P1-like PP carrying a gene, JL22, which was isolated from a clinical strain of Escherichia coli from a duck farm. P1-like PPs were very similar and conserved based on gene content similarities, with only eight highly variable regions. Importantly, two kinds of replicon types, namely, IncY and p0111, were identified and can be used to specifically identify the P1-like phage. JL22 is similar to P1, acquiring an important foreign DNA fragment with two obvious features, namely, the plasmid replication gene (p0111) replacing the gene (IncY) and a 4,200-bp fragment mobilized by IS and IS and containing a gene and a gene encoding tryptophan synthase (indole salvaging). The JL22 phage could be induced but had no lytic capacities. However, a lysogenic recipient and intact structure of JL22 virions were observed, showing that the extended-spectrum β-lactamase gene was successfully transferred. Overall, conserved genes can be a good complement to improve the identification efficiency and accuracy in future screening for P1-like PPs. Moreover, the highly conserved structures may be important for their prevalence and dissemination. As a PP, P1 DNA exists as a low-copy-number plasmid and replicates autonomously with a lysogenization style. This unique mode of P1-like elements probably indicates a stable contribution to antibiotic resistance. After analyzing these elements, we show that P1-like PPs are very similar and conserved, with only eight highly variable regions. Moreover, we observed the occurrence of replicon IncY and p0111 only in the P1-like PP community, implying that these conserved regions, coupled with IncY and p0111, can be an important complement in future screening of P1-like PPs. Identification and characterization of JL22 confirmed our findings that major changes were located in variable regions, including the first detection of in such a mobile genetic element. This suggests that these variable regions may facilitate foreign DNA mobilization. This study features a comprehensive genetic analysis of P1-like PPs, providing new insights into the dissemination mechanisms of antibiotic resistance through P1 PPs.
P1 样噬菌体质粒 (PP) 已被广泛用作分子生物学工具,但它作为一种活跃的辅助货物元素的作用尚未完全被理解。在这项研究中,我们深入了解了 RefSeq 数据库中 77 个 P1 样 PP 的结构特征和基因内容相似性。我们还描述了一个携带基因的 P1 样 PP,JL22,它是从一个鸭场的大肠杆菌临床株中分离出来的。P1 样 PP 基于基因内容相似性非常相似且保守,只有八个高度可变区域。重要的是,我们鉴定了两种复制子类型,即 IncY 和 p0111,可以用来特异性地识别 P1 样噬菌体。JL22 与 P1 相似,获得了一个重要的外来 DNA 片段,具有两个明显的特征,即质粒复制基因 (p0111) 取代基因 (IncY) 和一个由 IS 和 IS 移动的 4200bp 片段,其中包含基因和编码色氨酸合成酶(吲哚回收)的基因。JL22 噬菌体可以被诱导,但没有裂解能力。然而,观察到溶原性受体和 JL22 病毒粒子的完整结构,表明成功转移了扩展谱β-内酰胺酶基因。总的来说,保守基因可以很好地补充,以提高未来筛选 P1 样 PP 的效率和准确性。此外,高度保守的结构可能对它们的流行和传播很重要。作为一个 PP,P1 DNA 以低拷贝数质粒的形式存在,并以溶原化的方式自主复制。这种独特的 P1 样元素模式可能表明它对抗生素耐药性的稳定贡献。在分析这些元素后,我们表明 P1 样 PP 非常相似和保守,只有八个高度可变区域。此外,我们仅在 P1 样 PP 群落中观察到复制子 IncY 和 p0111 的发生,这意味着这些保守区域与 IncY 和 p0111 一起,可以成为未来筛选 P1 样 PP 的重要补充。JL22 的鉴定和表征证实了我们的发现,即主要变化位于可变区域,包括首次在这种可移动遗传元件中检测到。这表明这些可变区域可能促进了外源 DNA 的移动。本研究对 P1 样 PP 进行了全面的遗传分析,为通过 P1 PP 了解抗生素耐药性的传播机制提供了新的见解。