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- Title
Novel lnu(G) gene conferring resistance to lincomycin by nucleotidylation, located on Tn6260 from Enterococcus faecalis E531.
- Authors
Xiao-Qing Zhu; Xiao-Ming Wang; Hui Li; Yan-Hong Shang; Yu-Shan Pan; Cong-Ming Wu; Yang Wang; Xiang-Dang Du; Jian-Zhong Shen; Zhu, Xiao-Qing; Wang, Xiao-Ming; Li, Hui; Shang, Yan-Hong; Pan, Yu-Shan; Wu, Cong-Ming; Wang, Yang; Du, Xiang-Dang; Shen, Jian-Zhong
- Abstract
<bold>Objectives: </bold>To identify a novel putative lincosamide resistance gene determinant in a swine Enterococcus faecalis E531 exhibiting a lincosamide resistance/macrolide susceptibility (L R M S ) phenotype and to determine its location and genetic environment.<bold>Methods: </bold>The whole genomic DNA of E. faecalis E531, which tested negative for the known lincosamide nucleotidyltransferase genes, was sequenced. A putative lincosamide resistance gene determinant was cloned into an Escherichia coli - E. faecalis shuttle vector (pAM401) and transformed into E. faecalis JH2-2. The MICs were determined by the microbroth dilution method. Inactivity of lincomycin was examined by UPLC-MS/MS. Inverse PCR and primer walking were used to explore the genetic environment based on the assembled sequence.<bold>Results: </bold>A novel resistance gene, designated lnu (G), which encodes a putative lincosamide nucleotidyltransferase, was found in E. faecalis E531. The deduced Lnu(G) amino acid sequence displayed 76.0% identity to Lnu(B) in Enterococcus faecium . Both E. faecalis E531 and E. faecalis JH2-2 harbouring pAM401- lnu (G) showed a 4-fold increase in the MICs of lincomycin, compared with E. faecalis JH2-2 or E. faecalis JH2-2 harbouring empty vector pAM401 only. UPLC-MS/MS demonstrated that the Lnu(G) enzyme catalysed adenylylation of lincomycin. The genetic environment analysis revealed that the lnu (G) gene was embedded into a novel putative transposon, designated Tn 6260 , which was active.<bold>Conclusions: </bold>A novel lincosamide nucleotidyltransferase gene lnu (G) was identified in E. faecalis . The location of the lnu (G) gene on a mobile element Tn 6260 makes it easy to disseminate.
- Subjects
LINCOMYCIN; DRUG resistance in bacteria; ENTEROCOCCUS faecalis; NUCLEOTIDYLTRANSFERASES; AMINO acids; ANIMAL experimentation; ANTIBIOTICS; COMPARATIVE studies; DNA; DRUG resistance in microorganisms; ENTEROCOCCUS; GENOMES; RESEARCH methodology; MEDICAL cooperation; MICROBIAL sensitivity tests; RESEARCH; SWINE; GRAM-positive bacterial infections; EVALUATION research; SEQUENCE analysis; PHARMACODYNAMICS
- Publication
Journal of Antimicrobial Chemotherapy (JAC), 2017, Vol 72, Issue 4, p993
- ISSN
0305-7453
- Publication type
journal article
- DOI
10.1093/jac/dkw549