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食品及食品加工环境中分离的无致病性李斯特菌属中质粒携带的苯扎氯铵抗性盒 bcrABC 和镉抗性 cadA 基因的流行率。

Prevalence of plasmid-borne benzalkonium chloride resistance cassette bcrABC and cadmium resistance cadA genes in nonpathogenic Listeria spp. isolated from food and food-processing environments.

机构信息

Department of Applied Microbiology, Faculty of Biology, University of Warsaw, Miecznikowa 1, 02-096 Warsaw, Poland.

Department of Bacterial Genetics, Faculty of Biology, University of Warsaw, Miecznikowa 1, 02-096 Warsaw, Poland.

出版信息

Int J Food Microbiol. 2019 Feb 2;290:247-253. doi: 10.1016/j.ijfoodmicro.2018.10.019. Epub 2018 Oct 26.

Abstract

The sixty-seven nonpathogenic Listeria spp. strains isolated from food and food processing environments in Poland were examined for the presence of benzalkonium chloride (BC) resistance cassette (bcrABC) and four different variants of cadmium resistance determinants (cadA1-cadA4). All the strains were phenotypically resistant to cadmium and 22 among them were also resistant to BC. PCR-based analysis revealed that bcrABC cassette was harbored by 95.5% of the strains phenotypically resistant to BC. All of them harbored also either cadA1 or cadA2 genes (none carried cadA3 or cadA4), which corresponded to the presence of plasmids with two restriction patterns. The strains resistant to cadmium but susceptible to BC harbored only the cadA1 gene variant. DNA-DNA hybridization analysis showed that all the identified bcrABC, cadA1 and cadA2 genes were located within plasmids, classified into 11 groups of RFLP profiles. Only one of the plasmids - pLIS1 of Listeria welshimeri (carrying bcrABC and cadA2) - was capable of efficient conjugal transfer from nonpathogenic Listeria isolates to a pathogenic Listeria monocytogenes strain. Analysis of the complete nucleotide sequence of pLIS1 (the first sequenced plasmid of L. welshimeri species) revealed the presence of genes involved in plasmid replication, stabilization and transfer as well as genes conferring resistance phenotypes. Comparative analysis showed that pLIS1 genome is highly similar to a group of plasmids originating from L. monocytogenes strains. A common feature of pLIS1 and its relatives, besides the presence of the resistance genes, is the presence of numerous transposable elements (TEs). The analysis revealed the important role of TEs in both promoting genetic rearrangements within Listeria spp. plasmids and the acquisition of resistance determinants.

摘要

从波兰食品和食品加工环境中分离出的 67 株非致病性李斯特菌,对其苯扎氯铵(BC)抗性盒(bcrABC)和四种不同变体的镉抗性决定簇(cadA1-cadA4)进行了检测。所有菌株均表现出对镉的表型抗性,其中 22 株对 BC 也具有抗性。基于 PCR 的分析表明,表型上对 BC 具有抗性的 95.5%的菌株都携带 bcrABC 盒。它们还携带 cadA1 或 cadA2 基因(均未携带 cadA3 或 cadA4),这与具有两种限制图谱的质粒存在相对应。对镉具有抗性但对 BC 敏感的菌株仅携带 cadA1 基因变体。DNA-DNA 杂交分析表明,所有鉴定的 bcrABC、cadA1 和 cadA2 基因均位于质粒中,分为 11 组 RFLP 图谱。只有一种质粒 - 李斯特菌的 pLIS1(携带 bcrABC 和 cadA2) - 能够从非致病性李斯特菌分离株高效地转移到致病性李斯特菌单增李斯特菌菌株。对 pLIS1 的完整核苷酸序列分析(李斯特菌属物种的第一个测序质粒)表明,存在与质粒复制、稳定和转移以及赋予抗性表型的基因相关的基因。比较分析表明,pLIS1 基因组与一组源自李斯特菌单增李斯特菌菌株的质粒高度相似。pLIS1 及其亲缘质粒的一个共同特征是,除了存在抗性基因外,还存在大量转座元件(TEs)。分析表明,TEs 在促进李斯特菌属质粒内的遗传重排以及获得抗性决定簇方面发挥了重要作用。

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