Development of pVCR94ΔX from Vibrio cholerae, a prototype for studying multidrug resistant IncA/C conjugative plasmids.
Development of pVCR94ΔX from Vibrio cholerae, a prototype for studying multidrug resistant IncA/C conjugative plasmids.
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DOI：
10.3389/fmicb.2014.00044
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发表时间：
2014
影响因子：
5.2
通讯作者：
Burrus V
中科院分区：
文献类型：
--
作者：
Carraro N;Sauvé M;Matteau D;Lauzon G;Rodrigue S;Burrus V
在过去的几十年里，霍乱弧菌的抗生素耐药性稳步增长，成为受霍乱影响的国家的主要威胁。多药耐药（MDR）通过通常由SXT/R391家族的整合和接合元件（ICE）介导的横向基因转移在临床和环境霍乱弧菌菌株中传播。然而，在少数报道但似乎是孤立的病例中，霍乱弧菌的MDR显示与其他自我传播的遗传元件（如接合质粒）相关。印加/C接合质粒常与肠杆菌科细菌的多药耐药相关。到目前为止，印加/C质粒在霍乱弧菌或其他弧菌中还不常见。在这里，我们提出了一个详细的分析pVCR 94 ΔX来自pVCR 94，一种新的印加/C接合质粒中确定的霍乱弧菌临床菌株分离于1994年卢旺达霍乱爆发。发现pVCR 94赋予对磺胺甲恶唑、甲氧苄啶、氨苄青霉素、链霉素、四环素和氯霉素的抗性，并且以非常高的频率转移。序列分析显示其嵌合性以及负责转移和维持的核心基因与其他印加/C质粒和SXT/R391家族的ICE的高度相似性。虽然印加/C质粒被认为是抗生素耐药性的主要威胁，但它们的基础生物学很少受到关注，主要是因为难以遗传操纵这些MDR赋予元件。因此，我们开发了一个方便的衍生自pVCR 94，pVCR 94 Δ X，一个120.5-kb的接合质粒，仅编码磺胺甲恶唑抗性。使用pVCR 94 Δ X，我们确定了转移的起源（oriT），并发现了一个转移的必需基因，两者都位于共享的骨架内，允许所有印加/C质粒的注释更新。pVCR 94 Δ X可能是一个有用的模型，它将为印加/C接合质粒的基础生物学提供新的见解。
Antibiotic resistance has grown steadily in Vibrio cholerae over the last few decades to become a major threat in countries affected by cholera. Multi-drug resistance (MDR) spreads among clinical and environmental V. cholerae strains by lateral gene transfer often mediated by integrative and conjugative elements (ICEs) of the SXT/R391 family. However, in a few reported but seemingly isolated cases, MDR in V. cholerae was shown to be associated with other self-transmissible genetic elements such as conjugative plasmids. IncA/C conjugative plasmids are often found associated with MDR in isolates of Enterobacteriaceae. To date, IncA/C plasmids have not been commonly found in V. cholerae or other species of Vibrio. Here we present a detailed analysis of pVCR94ΔX derived from pVCR94, a novel IncA/C conjugative plasmid identified in a V. cholerae clinical strain isolated during the 1994 Rwandan cholera outbreak. pVCR94 was found to confer resistance to sulfamethoxazole, trimethoprim, ampicillin, streptomycin, tetracycline, and chloramphenicol and to transfer at very high frequency. Sequence analysis revealed its mosaic nature as well as high similarity of the core genes responsible for transfer and maintenance with other IncA/C plasmids and ICEs of the SXT/R391 family. Although IncA/C plasmids are considered a major threat in antibiotics resistance, their basic biology has received little attention, mostly because of the difficulty to genetically manipulate these MDR conferring elements. Therefore, we developed a convenient derivative from pVCR94, pVCR94Δ X, a 120.5-kb conjugative plasmid which only codes for sulfamethoxazole resistance. Using pVCR94Δ X, we identified the origin of transfer (oriT) and discovered an essential gene for transfer, both located within the shared backbone, allowing for an annotation update of all IncA/C plasmids. pVCR94Δ X may be a useful model that will provide new insights on the basic biology of IncA/C conjugative plasmids.
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影响因子：
4.9
作者：
Giske, Christian G.;Froding, Inga;Walsh, Timothy R.
通讯作者：
Walsh, Timothy R.
影响因子：
3.5
作者：
CHEREPANOV, PP;WACKERNAGEL, W
通讯作者：
WACKERNAGEL, W
影响因子：
4.5
作者：
Baharoglu Z;Bikard D;Mazel D
通讯作者：
Mazel D
影响因子：
3.2
作者：
Beaber, JW;Hochhut, B;Waldor, MK
通讯作者：
Waldor, MK
影响因子：
9.4
作者：
Dalsgaard, A;Forslund, A;Sandström, A
通讯作者：
Sandström, A