TABLE 1.

Bacterial strains and plasmids used in this study

Strain or plasmidCharacteristicsSource or Reference
K. pneumoniae KG914, KG1116, KG525, KG545, KG502, KG827Clinical isolates from neonatal specimensThis study
E. coli
    CSH-2metB F nalidixic acidr rifampinrT. Sawai, Chiba University
    XL1-BluesupE44 recA1 endA1 gyrA96 thi hsdR17(rK mK+) relA1 lac [F proAB+ lac1qZΔM15::Tn10(Tetr)]Stratagene
    BL21(DE3)pLysSF ompT hsdSB (rB mB) gal dcm (DE3) pLysS (Camr)Invitrogen
Plasmids
    pKGC525A natural plasmid carrying blaGES-3 of K. pneumoniae KG525This study
    pKGB525A recombinant plasmid carrying a 6.7-kb BamHI fragment containing blaGES-3 of K. pneumoniae KG525This study
    pKGM525A recombinant plasmid carrying a 11.6-kb BamHI fragment containing blaGES-3 of K. pneumoniae KG525This study
    pTAGES3A recombinant plasmid carrying a PCR fragment with the entire blaGES-3 sequence and its promoter region cloned into the pCR2.1 vectorThis study
    pGES3A recombinant plasmid carrying EcoR1 fragment from pTAGES3This study
    pIBC1A recombinant plamid carrying blaIBC-1 constructed from pGES3This study
    pBCSK+A cloning vector; chloramphenicolrStratagene
    pCR2.1A cloning vector; ampicillinr kanamycinrInvitrogen
    pET29a(+)An expression vector; kanamycinrNovagen
    pET-GES3A recombinant plasmid carrying PCR-amplified blaGES-3 gene ligated to pET29a(+)This study