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      Complete Nucleotide Sequence of CTX-M-15-Plasmids from Clinical Escherichia coli Isolates: Insertional Events of Transposons and Insertion Sequences

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          Abstract

          Background

          CTX-M-producing Escherichia coli strains are regarded as major global pathogens.

          Methodology/Principal Findings

          The nucleotide sequence of three plasmids (pEC_B24: 73801-bp; pEC_L8: 118525-bp and pEC_L46: 144871-bp) from Escherichia coli isolates obtained from patients with urinary tract infections and one plasmid (pEC_Bactec: 92970-bp) from an Escherichia coli strain isolated from the joint of a horse with arthritis were determined. Plasmid pEC_Bactec belongs to the IncI1 group and carries two resistance genes: bla TEM-1 and bla CTX-M-15. It shares more than 90% homology with a previously published bla CTX-M-plasmid from E. coli of human origin. Plasmid pEC_B24 belongs to the IncFII group whereas plasmids pEC_L8 and pEC_L46 represent a fusion of two replicons of type FII and FIA. On the pEC_B24 backbone, two resistance genes, bla TEM-1 and bla CTX-M-15, were found. Six resistance genes, bla TEM-1, bla CTX-M-15, bla OXA-1, aac6'-lb-cr, tetA and catB4, were detected on the pEC_L8 backbone. The same antimicrobial drug resistance genes, with the exception of tetA, were also identified on the pEC_L46 backbone. Genome analysis of all 4 plasmids studied provides evidence of a seemingly frequent transposition event of the bla CTX-M-15-IS Ecp1 element. This element seems to have a preferred insertion site at the tnpA gene of a bla TEM-carrying Tn 3-like transposon, the latter itself being inserted by a transposition event. The IS 26-composite transposon, which contains the bla OXA-1, aac6'-lb-cr and catB4 genes, was inserted into plasmids pEC_L8 and pEC_L46 by homologous recombination rather than a transposition event. Results obtained for pEC_L46 indicated that IS 26 also plays an important role in structural rearrangements of the plasmid backbone and seems to facilitate the mobilisation of fragments from other plasmids.

          Conclusions

          Collectively, these data suggests that IS 26 together with IS Ecp1 could play a critical role in the evolution of diverse multiresistant plasmids found in clinical Enterobacteriaceae.

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          Most cited references30

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          Resistance plasmid families in Enterobacteriaceae.

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            Inactivation of antibiotics and the dissemination of resistance genes.

            J. Davies (1994)
            The emergence of multidrug-resistant bacteria is a phenomenon of concern to the clinician and the pharmaceutical industry, as it is the major cause of failure in the treatment of infectious diseases. The most common mechanism of resistance in pathogenic bacteria to antibiotics of the aminoglycoside, beta-lactam (penicillins and cephalosporins), and chloramphenicol types involves the enzymic inactivation of the antibiotic by hydrolysis or by formation of inactive derivatives. Such resistance determinants most probably were acquired by pathogenic bacteria from a pool of resistance genes in other microbial genera, including antibiotic-producing organisms. The resistance gene sequences were subsequently integrated by site-specific recombination into several classes of naturally occurring gene expression cassettes (typically "integrons") and disseminated within the microbial population by a variety of gene transfer mechanisms. Although bacterial conjugation once was believed to be restricted in host range, it now appears that this mechanism of transfer permits genetic exchange between many different bacterial genera in nature.
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              The 3'-terminal sequence of Escherichia coli 16S ribosomal RNA: complementarity to nonsense triplets and ribosome binding sites.

              With a stepwise degradation and terminal labeling procedure the 3'-terminal sequence of E. coli 16S ribosomal RNA is shown to be Pyd-A-C-C-U-C-C-U-U-A(OH). It is suggested that this region of the RNA is able to interact with mRNA and that the 3'-terminal U-U-A(OH) is involved in the termination of protein synthesis through base-pairing with terminator codons. The sequence A-C-C-U-C-C could recognize a conserved sequence found in the ribosome binding sites of various coliphage mRNAs; it may thus be involved in the formation of the mRNA.30S subunit complex.
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                Author and article information

                Contributors
                Role: Editor
                Journal
                PLoS One
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, USA )
                1932-6203
                2010
                18 June 2010
                : 5
                : 6
                : e11202
                Affiliations
                [1 ]Department of Pathology, Bacteriology and Avian Diseases, Faculty of Veterinary Medicine, Ghent University, Merelbeke, Belgium
                [2 ]Laboratory of Pharmaceutical Biotechnology, Ghent University, Ghent, Belgium
                [3 ]NXTGNT sequencing facility, Ghent University, Ghent, Belgium
                [4 ]Biobix-Laboratory for Bioinformatics and Computational Genomics, Department of Molecular Biotechnology, Ghent University, Ghent, Belgium
                [5 ]Department of Bacteriology and Immunology, CODA-CERVA-VAR, Brussels, Belgium
                National Institute of Allergy and Infectious Diseases, National Institutes of Health, United States of America
                Author notes

                Conceived and designed the experiments: AS FVN. Performed the experiments: AS. Analyzed the data: AS TV. Contributed reagents/materials/analysis tools: FVN. Wrote the paper: AS. Provided important suggestions: AM DD PB FH. Supervised the whole project: AM DD PB FH.

                ¶ These authors also contributed equally to this work.

                Article
                10-PONE-RA-17525R1
                10.1371/journal.pone.0011202
                2887853
                20585456
                f15dbafa-3d85-4de0-888a-31d5eff0e219
                Smet et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
                History
                : 30 March 2010
                : 29 May 2010
                Page count
                Pages: 8
                Categories
                Research Article
                Biotechnology/Applied Microbiology
                Genetics and Genomics/Bioinformatics
                Genetics and Genomics/Comparative Genomics
                Genetics and Genomics/Microbial Evolution and Genomics
                Microbiology/Applied Microbiology
                Microbiology/Microbial Evolution and Genomics
                Infectious Diseases/Antimicrobials and Drug Resistance
                Infectious Diseases/Bacterial Infections
                Public Health and Epidemiology/Infectious Diseases

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                Uncategorized

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