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. 2017 Jul 25;61(8):e00811-17.
doi: 10.1128/AAC.00811-17. Print 2017 Aug.

Frequent Tn 2 Misannotation in the Genetic Background of rmtB

Willames M B S Martins  1 , Ana C Gales  2
Affiliations

Frequent Tn 2 Misannotation in the Genetic Background of rmtB

Willames M B S Martins et al. Antimicrob Agents Chemother. .
No abstract available

Keywords: 16S rRNA methyltransferase; Tn2; plasmids; transposons.

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Figures

FIG 1
FIG 1
(A) Comparison of the main transposon sequence regions (tnpA, res, tnpR, and blaTEM) with Tn3. Genes are shown by labeled arrows, inverted repeats of Tn3 are shown as black triangles, and the resolution site is a dark-gray box. Inverted repeats are located in the same positions of Tn2. Vertical black lines indicate differences in the nucleotide sequences of Tn3 (reference sequence) (GenBank accession number HM749966) from those of the other analyzed transposons. The red box in pCT-KPC and the p477Kp transposons indicate the presence of truncated tnpR. Insertion sequences were identified by ISfinder and are demonstrated by light-purple arrows. The pEC012 plasmid shows an insertion of a partial IS26 (ΔIS26) sequence and an IS15DI sequence that show 99% similarity with IS26, according to ISfinder. Purple arrows indicate the presence of rmtB downstream of Tn2 transposons in pHeBE7 (Tn2), pCT-KPC (ΔTn2), p477Kp (ΔTn2), pCC1409-1 (ΔTn2), and pEC012 (Tn2). This figure was compiled from alignments created using SeaView version 4.6.1. (B) In silico MslI restriction analysis of Tn2 and its variants proposed by Bailey and colleagues (8). Lane 1, Tn2 of pHeBE7; lane 2, Tn2 of pIP1206; lane 3, Tn2 of pMC-NDM; lane 4, ΔTn2 (tnpR and blaTEM-1b) of pC06114; lane 5, ΔTn2 of p397Kp (ΔtnpR and blaTEM-1b). In both ΔTn2 sequences analyzed, tnpA was missing. MslI restriction confirmed the presence of Tn2 in isolates with the complete transposon present (tnpA, tnpR, and blaTEM-1b). However, this methodology was shown to not be accurate in identifying the ΔTn2 sequence, since the digested fragment comprises the end of tnpA (absent in ΔTn2) and the beginning of blaTEM-1b. MW, molecular weight markers.
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