ena-STUDY-CARDIFF UNIVERSITY-31-08-2014-21:18:04:727-9
Whole genome sequence of Acinetobacter bereziniae CHI-40-1, harbouring plasmid pNDM-40-1.
A clinical isolate of Acinetobacter bereziniae, CHI-40-1, isolated in India in 2005 was fully sequenced using an an Illumina HiSeq platform. Reads were assembled de novo using the Velvet Assembly Tool (Version 1.2.10). Annotation was performed using RAST (Version 2.0). The genetic context of the metallo-ß-lactamase gene blaNDM-1 was defined, and the full plasmid pNDM-40-1 was closed by bridging of contig gaps using PCR and Sanger sequencing of amplicons. pNDM-40-1 was subjected to detailed comparison to similar plasmids and other blaNDM-1 contexts, and the annotation updated with reference to these related sequences. pNDM-40-1 was successfully transferred to Escherichia coli and Acinetobacter pittii recipients. The stability of the plasmid in A. bereziniae CHI-40-1 and transconjugants was explored by a passage experiment with and without exposure to meropenem. The presence of pNDM-40-1 was quantified by gene probing of S1 PFGE gels and quantitative real-time PCR.
A clinical isolate of Acinetobacter bereziniae, CHI-40-1, isolated in India in 2005 was fully sequenced using an an Illumina HiSeq platform. Reads were assembled de novo using the Velvet Assembly Tool (Version 1.2.10). Annotation was performed using RAST (Version 2.0). The genetic context of the metallo-ß-lactamase gene blaNDM-1 was defined, and the full plasmid pNDM-40-1 was closed by bridging of contig gaps using PCR and Sanger sequencing of amplicons. pNDM-40-1 was subjected to detailed comparison to similar plasmids and other blaNDM-1 contexts, and the annotation updated with reference to these related sequences. pNDM-40-1 was successfully transferred to Escherichia coli and Acinetobacter pittii recipients. The stability of the plasmid in A. bereziniae CHI-40-1 and transconjugants was explored by a passage experiment with and without exposure to meropenem. The presence of pNDM-40-1 was quantified by gene probing of S1 PFGE gels and quantitative real-time PCR.