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mecA 和 blaZ 调节元件在耐甲氧西林金黄色葡萄球菌中间型区域克隆株 mecA 表达中的作用。

The role of mecA and blaZ regulatory elements in mecA expression by regional clones of methicillin-resistant Staphylococcus pseudintermedius.

机构信息

Department of Comparative Medicine, College of Veterinary Medicine, University of Tennessee, 2407 River Drive, Knoxville, TN 37996, United States.

出版信息

Vet Microbiol. 2011 Aug 5;151(3-4):345-53. doi: 10.1016/j.vetmic.2011.03.026. Epub 2011 Mar 30.

Abstract

Two major regional clones of methicillin-resistant Staphylococcus pseudintermedius (MRSP) have been identified in Europe and North America. They are designated multilocus sequence types (ST) 71 and 68 and contain staphylococcal chromosome cassette (SCCmec) types II-III and V(T), respectively. One notable difference between the two clones is a deletion in the mecI/mecR1 regulatory apparatus of ST 68 SCCmec V(T). This deletion in analogous methicillin-resistant Staphylococcus aureus (MRSA) results in more responsive and greater expression of the mecA encoded penicillin-binding protein 2a, and is associated with SCCmec types occurring in community-acquired MRSA lineages. The aim of this study was to characterize mec and bla regulatory apparatuses in MRSP and determine their effects on expression of mecA. Seventeen S. pseudintermedius isolates representing nine methicillin-resistant ST lineages were screened for the presence of the repressors blaI and mecI and sensors blaR1 and mecR1. The bla and mec operons for each isolate were sequenced and compared for homology between the repressor open-reading frames (ORF), sensor ORFs, and mecA promoter regions. A real-time reverse transcriptase PCR expression assay was developed, validated and applied to nine isolates determining the effect of oxacillin induction on mecA transcription. Significant differences were found in mecA expression between isolates with a full regulatory complement (mecI/mecR1 and blaI/blaR1) and those with truncated and/or absent regulatory elements. Isolates representative of European and North American MRSP ST regional clones have dissimilar mecA responses to oxacillin.

摘要

耐甲氧西林金黄色葡萄球菌(MRSA)的两个主要区域克隆已在欧洲和北美被鉴定。它们被指定为多位点序列类型(ST)71 和 68,分别包含葡萄球菌染色体盒(SCCmec)类型 II-III 和 V(T)。这两个克隆之间的一个显著区别是 ST68 SCCmec V(T)的 mecI/mecR1 调控装置中的缺失。这种缺失在类似的耐甲氧西林金黄色葡萄球菌(MRSA)中导致 mecA 编码的青霉素结合蛋白 2a 的响应更敏感和表达更大,并且与社区获得性 MRSA 谱系中发生的 SCCmec 类型相关。本研究的目的是表征 MRSP 中的 mec 和 bla 调控装置,并确定它们对 mecA 表达的影响。筛选了代表 9 种耐甲氧西林 ST 谱系的 17 株中间葡萄球菌分离株,以检测存在 blaI 和 mecI 抑制剂和 blaR1 和 mecR1 传感器。对每个分离株的 bla 和 mec 操纵子进行了测序,并比较了抑制物开放阅读框(ORF)、传感器 ORF 和 mecA 启动子区域之间的同源性。开发、验证并应用了实时逆转录 PCR 表达测定法,以确定 9 株分离株中 mecA 转录对苯唑西林诱导的影响。在具有完整调控因子(mecI/mecR1 和 blaI/blaR1)的分离株和具有截短和/或缺失调控元件的分离株之间,发现 mecA 表达存在显著差异。代表欧洲和北美 MRSP ST 区域克隆的分离株对苯唑西林的 mecA 反应不同。

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