{"title":"多粘菌素 B 诱导的沙门氏菌耐药突变体中的长目标重复插入。","authors":"","doi":"10.1016/j.jgar.2024.07.002","DOIUrl":null,"url":null,"abstract":"<div><h3>Objectives</h3><p>A <em>Salmonella enterica</em> subsp. <em>diarizonae</em> (hereafter <em>S. diarizonae</em>) clinical strain S499 demonstrated unique genomic features. The strain S499 was treated with polymyxin B in vitro to investigate the mechanism of resistance.</p></div><div><h3>Methods</h3><p>S499 was treated with polymyxin B by increasing concentration gradually to obtain a resistant mutant S499V. Whole genomes of the two strains were sequenced using Illumina HiSeq X<strong>-</strong>10 and PacBio RS II platforms. Reverse transcription–quantitative polymerase chain reaction (RT-qPCR) was performed to compare the gene expression.</p></div><div><h3>Results</h3><p>The chromosome of strain S499 contained a 40-kb DNA region that was replicated after treatment with polymyxin B and generated a triple tandem DNA repeat region in the chromosome of mutant strain S499V. This repeat region in S499V was flanked by IS<em>1</em> and contained <em>pmrD, pmrG</em>, and <em>arnBCADTEF</em> operon<em>.</em> In comparison to the homologous 40-kb DNA region of strain S499, a few genes in the repeat DNA region of strain S499V contained truncating mutations that generate two open reading frames (ORFs). The expression of <em>pmrD, pmrG</em>, and <em>arnT</em> was significantly upregulated in S499V.</p></div><div><h3>Conclusion</h3><p>The duplication and overexpression of <em>pmrD, pmrG</em>, and <em>arnT</em> operon may be responsible for the polymyxin B resistance of mutant strain S499V.</p></div>","PeriodicalId":15936,"journal":{"name":"Journal of global antimicrobial resistance","volume":null,"pages":null},"PeriodicalIF":3.7000,"publicationDate":"2024-07-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2213716524001334/pdfft?md5=b614bd4747185be867b00655dbb4f751&pid=1-s2.0-S2213716524001334-main.pdf","citationCount":"0","resultStr":"{\"title\":\"Insertion with long target duplication in polymyxin B-induced resistant mutant of Salmonella\",\"authors\":\"\",\"doi\":\"10.1016/j.jgar.2024.07.002\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><h3>Objectives</h3><p>A <em>Salmonella enterica</em> subsp. <em>diarizonae</em> (hereafter <em>S. diarizonae</em>) clinical strain S499 demonstrated unique genomic features. The strain S499 was treated with polymyxin B in vitro to investigate the mechanism of resistance.</p></div><div><h3>Methods</h3><p>S499 was treated with polymyxin B by increasing concentration gradually to obtain a resistant mutant S499V. Whole genomes of the two strains were sequenced using Illumina HiSeq X<strong>-</strong>10 and PacBio RS II platforms. Reverse transcription–quantitative polymerase chain reaction (RT-qPCR) was performed to compare the gene expression.</p></div><div><h3>Results</h3><p>The chromosome of strain S499 contained a 40-kb DNA region that was replicated after treatment with polymyxin B and generated a triple tandem DNA repeat region in the chromosome of mutant strain S499V. This repeat region in S499V was flanked by IS<em>1</em> and contained <em>pmrD, pmrG</em>, and <em>arnBCADTEF</em> operon<em>.</em> In comparison to the homologous 40-kb DNA region of strain S499, a few genes in the repeat DNA region of strain S499V contained truncating mutations that generate two open reading frames (ORFs). The expression of <em>pmrD, pmrG</em>, and <em>arnT</em> was significantly upregulated in S499V.</p></div><div><h3>Conclusion</h3><p>The duplication and overexpression of <em>pmrD, pmrG</em>, and <em>arnT</em> operon may be responsible for the polymyxin B resistance of mutant strain S499V.</p></div>\",\"PeriodicalId\":15936,\"journal\":{\"name\":\"Journal of global antimicrobial resistance\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":3.7000,\"publicationDate\":\"2024-07-14\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://www.sciencedirect.com/science/article/pii/S2213716524001334/pdfft?md5=b614bd4747185be867b00655dbb4f751&pid=1-s2.0-S2213716524001334-main.pdf\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Journal of global antimicrobial resistance\",\"FirstCategoryId\":\"3\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/S2213716524001334\",\"RegionNum\":3,\"RegionCategory\":\"医学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q2\",\"JCRName\":\"INFECTIOUS DISEASES\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of global antimicrobial resistance","FirstCategoryId":"3","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S2213716524001334","RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"INFECTIOUS DISEASES","Score":null,"Total":0}
引用次数: 0
摘要
目的：肠炎沙门氏菌 diarizonae 亚种（以下简称 diarizonae）临床菌株 S499 具有独特的基因组特征。在体外用多粘菌素 B 处理 S499 菌株，以研究其耐药性机制：方法：用多粘菌素 B 处理 S499，逐渐增加浓度，以获得抗性突变体 S499V。使用 Illumina HiSeq X-10 和 PacBio RS II 平台对两株菌株的全基因组进行测序。对基因表达进行了反转录定量 PCR（RT-qPCR）比较：结果：菌株 S499 的染色体包含一个 40 kb 的 DNA 区域，经多粘菌素 B 处理后复制，在突变菌株 S499V 的染色体中产生了一个三重串联 DNA 重复区域。S499V 的这一重复区两侧是 IS1，包含 pmrD、pmrG 和 arnBCADTEF 操作子。与菌株 S499 的同源 40-kb DNA 区域相比，菌株 S499V 重复 DNA 区域中的一些基因含有截短突变，产生了两个开放阅读框（ORF）。在 S499V 中，pmrD、pmrG 和 arnT 的表达显著上调：结论：pmrD、pmrG 和 arnT 操作子的重复和过度表达可能是突变株 S499V 对多粘菌素 B 产生抗性的原因。
Insertion with long target duplication in polymyxin B-induced resistant mutant of Salmonella
Objectives
A Salmonella enterica subsp. diarizonae (hereafter S. diarizonae) clinical strain S499 demonstrated unique genomic features. The strain S499 was treated with polymyxin B in vitro to investigate the mechanism of resistance.
Methods
S499 was treated with polymyxin B by increasing concentration gradually to obtain a resistant mutant S499V. Whole genomes of the two strains were sequenced using Illumina HiSeq X-10 and PacBio RS II platforms. Reverse transcription–quantitative polymerase chain reaction (RT-qPCR) was performed to compare the gene expression.
Results
The chromosome of strain S499 contained a 40-kb DNA region that was replicated after treatment with polymyxin B and generated a triple tandem DNA repeat region in the chromosome of mutant strain S499V. This repeat region in S499V was flanked by IS1 and contained pmrD, pmrG, and arnBCADTEF operon. In comparison to the homologous 40-kb DNA region of strain S499, a few genes in the repeat DNA region of strain S499V contained truncating mutations that generate two open reading frames (ORFs). The expression of pmrD, pmrG, and arnT was significantly upregulated in S499V.
Conclusion
The duplication and overexpression of pmrD, pmrG, and arnT operon may be responsible for the polymyxin B resistance of mutant strain S499V.
期刊介绍：
The Journal of Global Antimicrobial Resistance (JGAR) is a quarterly online journal run by an international Editorial Board that focuses on the global spread of antibiotic-resistant microbes.
JGAR is a dedicated journal for all professionals working in research, health care, the environment and animal infection control, aiming to track the resistance threat worldwide and provides a single voice devoted to antimicrobial resistance (AMR).
Featuring peer-reviewed and up to date research articles, reviews, short notes and hot topics JGAR covers the key topics related to antibacterial, antiviral, antifungal and antiparasitic resistance.
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