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RamA, a transcriptional regulator conferring florfenicol resistance in Leclercia adecarboxylata R25
C. Cheng, Y. Ying, D. Zhou, L. Zhu, J. Lu, A. Li, Q. Bao, M. Zhu
Language English Country United States
Document type Journal Article
Grant support
Nos. LQ17H190001
Natural Science Foundation of Zhejiang Province
LY19C060002
Natural Science Foundation of Zhejiang Province
- MeSH
- Anti-Bacterial Agents pharmacology MeSH
- Bacterial Proteins genetics metabolism MeSH
- Enterobacteriaceae drug effects genetics metabolism MeSH
- Escherichia coli genetics MeSH
- Gene Knockout Techniques MeSH
- Cloning, Molecular MeSH
- Microbial Sensitivity Tests MeSH
- Drug Resistance, Multiple, Bacterial drug effects genetics MeSH
- RNA, Ribosomal, 16S MeSH
- Whole Genome Sequencing MeSH
- Thiamphenicol analogs & derivatives MeSH
- Trans-Activators genetics MeSH
- Computational Biology MeSH
- Publication type
- Journal Article MeSH
Due to the inappropriate use of florfenicol in agricultural practice, florfenicol resistance has become increasingly serious. In this work, we studied the novel florfenicol resistance mechanism of an animal-derived Leclercia adecarboxylata strain R25 with high-level florfenicol resistance. A random genomic DNA library was constructed to screen the novel florfenicol resistance gene. Gene cloning, gene knockout, and complementation combined with the minimum inhibitory concentration (MIC) detection were conducted to determine the function of the resistance-related gene. Sequencing and bioinformatics methods were applied to analyze the structure of the resistance gene-related sequences. Finally, we obtained a regulatory gene of an RND (resistance-nodulation-cell division) system, ramA, that confers resistance to florfenicol and other antibiotics. The ramA-deleted variant (LA-R25ΔramA) decreased the level of resistance against florfenicol and several other antibiotics, while a ramA-complemented strain (pUCP24-prom-ramA/LA-R25ΔramA) restored the drug resistance. The whole-genome sequencing revealed that there were five RND efflux pump genes (mdtABC, acrAB, acrD, acrEF, and acrAB-like) encoded over the chromosome, and ramA located upstream of the acrAB-like genes. The results of this work suggest that ramA confers resistance to florfenicol and other structurally unrelated antibiotics, presumably by regulating the RND efflux pump genes in L. adecarboxylata R25.
Department of Clinical Laboratory Zhejiang Hospital Hangzhou 310013 Zhejiang China
Medical Research Center Taizhou Hospital of Zhejiang Province Taizhou 317000 Zhejiang China
The 5th Affiliated Hospital Wenzhou Medical University Lishui 323000 Zhejiang China
Vocational and Technical College Lishui University Lishui 323000 China
References provided by Crossref.org
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- $a Due to the inappropriate use of florfenicol in agricultural practice, florfenicol resistance has become increasingly serious. In this work, we studied the novel florfenicol resistance mechanism of an animal-derived Leclercia adecarboxylata strain R25 with high-level florfenicol resistance. A random genomic DNA library was constructed to screen the novel florfenicol resistance gene. Gene cloning, gene knockout, and complementation combined with the minimum inhibitory concentration (MIC) detection were conducted to determine the function of the resistance-related gene. Sequencing and bioinformatics methods were applied to analyze the structure of the resistance gene-related sequences. Finally, we obtained a regulatory gene of an RND (resistance-nodulation-cell division) system, ramA, that confers resistance to florfenicol and other antibiotics. The ramA-deleted variant (LA-R25ΔramA) decreased the level of resistance against florfenicol and several other antibiotics, while a ramA-complemented strain (pUCP24-prom-ramA/LA-R25ΔramA) restored the drug resistance. The whole-genome sequencing revealed that there were five RND efflux pump genes (mdtABC, acrAB, acrD, acrEF, and acrAB-like) encoded over the chromosome, and ramA located upstream of the acrAB-like genes. The results of this work suggest that ramA confers resistance to florfenicol and other structurally unrelated antibiotics, presumably by regulating the RND efflux pump genes in L. adecarboxylata R25.
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