Table 1.

Strains, plasmids, and primers used in this study

Strain, plasmid, or primerRelevant genotype, description, or sequenceSource or reference
Strains
    E. coli Bre-1Clinical E. coli strain producing CTX-M-93 (pI 8.9)This study
    E. coli Top10supE44 hsdS20 (rB mB) recA13 ara-14 proA2 lacY1 galK2 rpsL20 xyl-5 mtl-1Stratagene
    E. coli J53Azide-resistant E. coli strain22
    E. coli 50192E. coli NTCC 50192 harboring 154-, 66-, 48-, and 7-kb plasmids22
Plasmids
    pNBRE-1Natural plasmid encoding CTX-M-93This study
    pPCRScriptRecombinant pPCRScript plasmidStratagene
    pET-9aRecombinant pET-9a plasmidStratagene
    pM-93Recombinant pPCRScript plasmid that contains a 0.9-kb fragment encoding CTX-M-93 (pI 8.9)This study
    pM-93NBRecombinant pPCRScript plasmid that contains a 0.9-kb fragment encoding CTX-M-93 with NdeI and BamHI restriction sitesThis study
    pM-27Recombinant pPCRScript plasmid that contains a 0.9-kb fragment encoding CTX-M-27 (Gln169Leu)This study
    pM-93-Q169MRecombinant pPCRScript plasmid that contains a 0.9-kb fragment encoding a modified CTX-M-93 protein with a Gln169Met substitutionThis study
    pM-27-L169CRecombinant pPCRScript plasmid that contains a 0.9-kb fragment encoding a modified CTX-M-27 protein with a Leu169Cys substitutionThis study
    pET-M-93Recombinant pET-9a plasmid that contains a 0.9-kb fragment encoding CTX-M-93This study
Primersa
    TEM-α5′-GAGTATTCAACATTTCCGTGT-3′24
    TEM-β5′-TAATCAGTGAGGCACCTATCT-3′24
    SHV-A5′-ATGCGTTATWTTCGCCTGTGT-3′24
    SHV-B5′-TTAGCGTTGCCAGTGCTCG-3′24
    CTX-M-A5′-CGCTTTGCGATGTGCAG-3′24
    CTX-M-B5′-ACCGCGATATCGTTGGT-3′24
    TLA-2A5′-TCCCTGGAGCACTTATGAAT-3′15
    TLA-2B5′-ATTAAGGATAAACTCATCCGC-3′15
    TLA-1A5′-GGCTAAAGGTACGGATTCGC-3′This study
    TLA-1B5′-ACGCTTTTGCAAATTTCGGC-3′This study
    SFO-A5′-TTACGTCAAACCACCCTGATGG-3′This study
    SFO-B5′-TTCTGCATTCTGCTGTGGCTG-3′This study
    BES-A5′-ATGTGGCAGTGGCTTGGAAA-3′This study
    BES-B5′-TTATCTTGCAGTACCAGTCG-3′This study
    BEL-1A5′-CGACAATGCCGCAGCTAACC-3′26
    BEL-1B5′-CAGAAGCAATTAATAACGCCC-3′26
    VEB-1A5′-CGACTTCCATTTCCCGATGC-3′21
    VEB-1B5′-GGACTCTGCAACAAATACGC-3′21
    PER-A5′-ATGAATGTCATTATAAAAGC-3′21
    PER-B5′-AATTTGGGCTTAGGGCAGAA-3′21
    GES-1A5′-ATGCGCTTCATTCACGCAC-3′28
    GES-1B5′-CTATTTGTCCGTGCTCAGG-3′28
    CTX-M-ATG5′-ATGGTGACAAAGAGAGTGCA-3′This study
    CTX-M-Stop5′-TTACAGCCCTTCGGCGATGA-3′This study
    CTX-M-NdeI5′-AAAACATATGGTGACAAAGAGAGTGCA-3′This study
    CTX-M-BamHI5′-AAAAGGATCCTTACAGCCCTTCGGCGATGA-3′This study
    ISEcp prom+5′-TGCTCTGTGGATAACTTGC-3′24
    IS903B15′-GGCTTTGTTGAATAAATCAG-3′24
    CTX-M-93-Q169L-A5′-GAATGGCGGTATTCAGCGTAGGTTCAG-3′This study
    CTX-M-93-Q169L-B5′-CTGAACCTACGCTGAATACCGCCATTC-3′This study
    CTX-M-93-Q169M-A5′-GAATGGCGGTATTCATCGTAGGTTCAG-3′This study
    CTX-M-93-Q169M-B5′-CTGAACCTACGATGAATACCGCCATTC-3′This study
    CTX-M-27-L169C-A5′-GAATGGCGGTATTACACGTAGGTTCAG-3′This study
    CTX-M-27-L169C-B5′-CTGAACCTACGTGTAATACCGCCATTC-3′This study
  • a Primers for Rep typing were reported previously (8). Primers for MLST typing were from the MLST databases at the University College Cork (http://mlst.ucc.ie/mlst/dbs/Ecoli) (37).