Plasmid-borne blaSHV genes in Klebsiella pneumoniae are associated with strong promoters

M TURNER, Patiyan Andersson, J Bell, J Turnidge, Tegan Harris, Philip Giffard

    Research output: Contribution to journalArticleResearchpeer-review

    Abstract

    Background Extended-spectrum ?-lactamases (ESBLs) belonging to the SHV family remain a major cause of ESBL-positive phenotypes in Klebsiella pneumoniae. The blaSHV gene is a normal constituent of the K. pneumoniae chromosome. However, most ESBL-encoding blaSHV genes found in K. pneumoniae are plasmid borne. The objective was to determine the contribution of promoter variants to the expression of plasmid-borne blaSHV genes. Methods: K. pneumoniae clinical isolates were analysed for the presence of IS26 insertions characteristic of plasmid-borne blaSHV, and differences in their blaSHV promoter sequences and expression levels. A high resolution melting (HRM)-based method for rapid promoter analysis was developed. Results: An IS26 insertion characteristic of the plasmid-borne blaSHV-1blaSHV-2/blaSHV-5 family was 100% linked to a promoter mutated in the 210 region, a mutation previously only found on the chromosome. The mutation was shown by real-time reverse transcriptase PCR to be associated with increased blaSHV expression. Conclusions: Plasmid-borne blaSHV is associated with strong promoters. It is likely that an SHV-dependent ESBL-positive phenotype requires both a strong promoter and a coding sequence mutation. An HRM assay can indicate blaSHV expression. � The Author 2009. Published by Oxford University Press on behalf of the British Society for Antimicrobial Chemotherapy. All rights reserved.
    Original languageEnglish
    Pages (from-to)960-964
    Number of pages5
    JournalJournal of Antimicrobial Chemotherapy
    Volume64
    Issue number5
    Publication statusPublished - 2009

    Fingerprint

    Klebsiella pneumoniae
    Plasmids
    Genes
    Mutation
    Freezing
    Chromosomes
    Phenotype
    Reverse Transcriptase Polymerase Chain Reaction
    Real-Time Polymerase Chain Reaction

    Cite this

    TURNER, M ; Andersson, Patiyan ; Bell, J ; Turnidge, J ; Harris, Tegan ; Giffard, Philip. / Plasmid-borne blaSHV genes in Klebsiella pneumoniae are associated with strong promoters. In: Journal of Antimicrobial Chemotherapy. 2009 ; Vol. 64, No. 5. pp. 960-964.
    @article{b990cc8ba3294dcb81a81063cf73081b,
    title = "Plasmid-borne blaSHV genes in Klebsiella pneumoniae are associated with strong promoters",
    abstract = "Background Extended-spectrum ?-lactamases (ESBLs) belonging to the SHV family remain a major cause of ESBL-positive phenotypes in Klebsiella pneumoniae. The blaSHV gene is a normal constituent of the K. pneumoniae chromosome. However, most ESBL-encoding blaSHV genes found in K. pneumoniae are plasmid borne. The objective was to determine the contribution of promoter variants to the expression of plasmid-borne blaSHV genes. Methods: K. pneumoniae clinical isolates were analysed for the presence of IS26 insertions characteristic of plasmid-borne blaSHV, and differences in their blaSHV promoter sequences and expression levels. A high resolution melting (HRM)-based method for rapid promoter analysis was developed. Results: An IS26 insertion characteristic of the plasmid-borne blaSHV-1blaSHV-2/blaSHV-5 family was 100{\%} linked to a promoter mutated in the 210 region, a mutation previously only found on the chromosome. The mutation was shown by real-time reverse transcriptase PCR to be associated with increased blaSHV expression. Conclusions: Plasmid-borne blaSHV is associated with strong promoters. It is likely that an SHV-dependent ESBL-positive phenotype requires both a strong promoter and a coding sequence mutation. An HRM assay can indicate blaSHV expression. � The Author 2009. Published by Oxford University Press on behalf of the British Society for Antimicrobial Chemotherapy. All rights reserved.",
    keywords = "beta lactamase SHV, extended spectrum beta lactamase, beta lactamase, article, bacterial chromosome, bacterial genetics, bacterium isolate, controlled study, gene expression, gene insertion, gene mutation, gene sequence, genetic analysis, genetic variability, Klebsiella pneumoniae, nonhuman, nucleotide sequence, phenotype, plasmid, promoter region, protein family, reverse transcription polymerase chain reaction, biosynthesis, enzymology, gene expression profiling, genetics, human, isolation and purification, Klebsiella infection, microbiology, molecular genetics, transposon, beta-Lactamases, DNA Transposable Elements, Gene Expression Profiling, Humans, Klebsiella Infections, Molecular Sequence Data, Plasmids, Promoter Regions, Genetic",
    author = "M TURNER and Patiyan Andersson and J Bell and J Turnidge and Tegan Harris and Philip Giffard",
    year = "2009",
    language = "English",
    volume = "64",
    pages = "960--964",
    journal = "Journal of Antimicrobial Chemotherapy",
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    TURNER, M, Andersson, P, Bell, J, Turnidge, J, Harris, T & Giffard, P 2009, 'Plasmid-borne blaSHV genes in Klebsiella pneumoniae are associated with strong promoters', Journal of Antimicrobial Chemotherapy, vol. 64, no. 5, pp. 960-964.

    Plasmid-borne blaSHV genes in Klebsiella pneumoniae are associated with strong promoters. / TURNER, M; Andersson, Patiyan; Bell, J; Turnidge, J; Harris, Tegan; Giffard, Philip.

    In: Journal of Antimicrobial Chemotherapy, Vol. 64, No. 5, 2009, p. 960-964.

    Research output: Contribution to journalArticleResearchpeer-review

    TY - JOUR

    T1 - Plasmid-borne blaSHV genes in Klebsiella pneumoniae are associated with strong promoters

    AU - TURNER, M

    AU - Andersson, Patiyan

    AU - Bell, J

    AU - Turnidge, J

    AU - Harris, Tegan

    AU - Giffard, Philip

    PY - 2009

    Y1 - 2009

    N2 - Background Extended-spectrum ?-lactamases (ESBLs) belonging to the SHV family remain a major cause of ESBL-positive phenotypes in Klebsiella pneumoniae. The blaSHV gene is a normal constituent of the K. pneumoniae chromosome. However, most ESBL-encoding blaSHV genes found in K. pneumoniae are plasmid borne. The objective was to determine the contribution of promoter variants to the expression of plasmid-borne blaSHV genes. Methods: K. pneumoniae clinical isolates were analysed for the presence of IS26 insertions characteristic of plasmid-borne blaSHV, and differences in their blaSHV promoter sequences and expression levels. A high resolution melting (HRM)-based method for rapid promoter analysis was developed. Results: An IS26 insertion characteristic of the plasmid-borne blaSHV-1blaSHV-2/blaSHV-5 family was 100% linked to a promoter mutated in the 210 region, a mutation previously only found on the chromosome. The mutation was shown by real-time reverse transcriptase PCR to be associated with increased blaSHV expression. Conclusions: Plasmid-borne blaSHV is associated with strong promoters. It is likely that an SHV-dependent ESBL-positive phenotype requires both a strong promoter and a coding sequence mutation. An HRM assay can indicate blaSHV expression. � The Author 2009. Published by Oxford University Press on behalf of the British Society for Antimicrobial Chemotherapy. All rights reserved.

    AB - Background Extended-spectrum ?-lactamases (ESBLs) belonging to the SHV family remain a major cause of ESBL-positive phenotypes in Klebsiella pneumoniae. The blaSHV gene is a normal constituent of the K. pneumoniae chromosome. However, most ESBL-encoding blaSHV genes found in K. pneumoniae are plasmid borne. The objective was to determine the contribution of promoter variants to the expression of plasmid-borne blaSHV genes. Methods: K. pneumoniae clinical isolates were analysed for the presence of IS26 insertions characteristic of plasmid-borne blaSHV, and differences in their blaSHV promoter sequences and expression levels. A high resolution melting (HRM)-based method for rapid promoter analysis was developed. Results: An IS26 insertion characteristic of the plasmid-borne blaSHV-1blaSHV-2/blaSHV-5 family was 100% linked to a promoter mutated in the 210 region, a mutation previously only found on the chromosome. The mutation was shown by real-time reverse transcriptase PCR to be associated with increased blaSHV expression. Conclusions: Plasmid-borne blaSHV is associated with strong promoters. It is likely that an SHV-dependent ESBL-positive phenotype requires both a strong promoter and a coding sequence mutation. An HRM assay can indicate blaSHV expression. � The Author 2009. Published by Oxford University Press on behalf of the British Society for Antimicrobial Chemotherapy. All rights reserved.

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    KW - gene sequence

    KW - genetic analysis

    KW - genetic variability

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    KW - beta-Lactamases

    KW - DNA Transposable Elements

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    KW - Klebsiella Infections

    KW - Molecular Sequence Data

    KW - Plasmids

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