TABLE 1.

Bacterial strains and plasmids used in this study

Strain/plasmidRelevant characteristic(s)aSource or reference
A. baumannii
    ATCC 19606TClinical isolate, type strainATCC
    ATCC 17978Clinical isolateATCC
    ATCC 17978.ORblsA::aph derivative of 17978; KmrThis work
    ATCC 17978.ORc17978.OR harboring pWHBLSA; Kmr AmprThis work
    ATCC 17978.ORp17978.OR harboring pWH1266; Kmr Tetr AmprThis work
    AYEClinical isolateATCC
    LUH 07672Clinical isolate, EU clone III47
    LUH 8809Clinical isolate, EU clone I48
    LUH 5875Clinical isolate, reference strain, EU clone III51
    LUH 13000Clinical isolate, EU clone IIL. Dijkshoorn
    RUH 134Clinical isolate, reference strain, EU clone II13
    RUH 875Clinical isolate, reference strain, EU clone I13
C. albicans tup1Constitutive filamentous derivative of SC53149
E. coli
    DH5αUsed for DNA recombinant methodsGibco-BRL
    Top10Used for DNA recombinant methodsInvitrogen
    EC100D+pir+, host for pKNOCK-Amp maintenanceEpicentre
    HB101Conjugation helper strain harboring pRK2073; Tpr Str7
    BL21 (DE)Overexpression of His-tagged BlsANovagen
Plasmids
    pCR-Blunt II-TOPOPCR cloning vector; Kmr ZeorInvitrogen
    pGEM-T EasyPCR cloning vector; AmprPromega
    pUC4KSource of the Kmr cassette; Kmr AmprPharmacia Biotech
    pET-TEVpET28a (Novagen) containing TEV protease cleavage site; Kmr25
    pKNOCK-AmpSuicide vector for allelic exchange; Ampr2
    pRK2073Used as helper in plasmid conjugation; Tpr29
    pWH1266E. coli-A. baumannii shuttle vector; Ampr Tcr26
    pBLSAblsA cloned into pCR-Blunt II-TOPO; Kmr ZeorThis work
    pKABLSApKNOCK-Amp harboring blsA; AmprThis work
    pKABLSA-KmpKABLSA with the pUC4K Kmr cassette inserted into NsiI restriction site; Ampr KmrThis work
    pGBLSA1Amplicon harboring blsA promoter and coding region cloned into pGEM-T Easy; AmprThis work
    pWHBLSApWH1266 harboring wild-type copy of blsA expressed under its own promoter; AmprThis work
    pGBLSA2Amplicon harboring blsA coding region cloned into pGEM-T Easy; AmprThis work
    pEBLSApET-TEV harboring a parental copy of blsA; KmrThis work
  • a Ampr, ampicillin resistance; Kmr, kanamycin resistance; Str, streptomycin resistance; Tcr, tetracycline resistance; Tpr, trimethoprim resistance; Zeor, zeocin resistance.