TABLE 1.
Strains and plasmids used in the study
| Strain or plasmid | Genotype/relevant characteristic(s)a | Reference |
|---|---|---|
| Strains | ||
| E. asburiae Temo_S | Human clinical isolate Ecc20200302 (isolated from urine, 28 February 2020); Tems | This study |
| E. asburiae Temo_R | Human clinical isolate Ecc20200303 (isolated from urine, 28 February 2020); Temr | This study |
| Temo_S/pBAD | E. asburiae Temo_S trans-complemented strain carrying pBAD202/D-TOPO | This study |
| Temo_S/pBADΩmdtABCD | E. asburiae Temo_S trans-complemented strain carrying pBAD202/D-TOPOΩmdtABCD (ECL_03401-ECL_03404) | This study |
| Temo_S/pBADΩacrD | E. asburiae Temo_S trans-complemented strain carrying pBAD202/D-TOPOΩacrD (ECL_03767) | This study |
| E. cloacae subsp. cloacae ATCC 13047 | Reference strain ATCC 13047 (cluster XI) | 51 |
| ATCC 13047ΔbaeS | ATCC 13047 deleted for baeS (ECL_03405) | This study |
| ATCC 13047ΔmdtB | ATCC 13047 deleted for mdtB (ECL_03403) | 63 |
| ATCC 13047ΔacrD | ATCC 13047 deleted for acrD (ECL_03767) | 63 |
| ATCC 13047/pBAD | ATCC 13047 trans-complemented strain carrying pBAD202/D-TOPO | 63 |
| ATCC 13047/pBADΩmdtABCD | ATCC 13047 trans-complemented strain carrying pBAD202/D-TOPOΩmdtABCD (ECL_03401-ECL_03404) | 63 |
| ATCC 13047/pBADΩacrD | ATCC 13047 trans-complemented strain carrying pBAD202/D-TOPOΩacrD (ECL_03767) | 63 |
| E. coli CFT073 | Wild-type susceptible clinical isolate (phylogenetic group B2) | 52 |
| CFT073_baeSThr175Pro | CFT073 derivative with allelic replacement of baeS by baeSThr175Pro | This study |
| E. coli DH5α λpir | Reference strain (supE4 ΔlacU169 [Φ80 lacZΔM15] hsdR17 recA1 endA1 gyrA96 thi-1 relA1 λpir) | Laboratory collection |
| Plasmids | ||
| pBAD202/D-TOPO | General expression vector with arabinose-inducible promoter; Kanr | Life Technologies |
| pKOBEG | Recombination vector, phage λ recγβα operon under control of pBAD promoter; Cmr | 60 |
| pKD4 | Plasmid containing FRT-flanked kanamycin cassette; Kanr | 59 |
| pCP20_Gm | Flp-mediated recombination vector; Gmr | 61 |
| pDS132 | Conditional replication vector, R6K origin, mobRP4 transfer origin, sucrose-inducible sacB, Cmr | 64 |
a
Tems, temocillin susceptible; Temr, temocillin resistant; Cmr, chloramphenicol resistant; Gmr, gentamicin resistant; Kanr, kanamycin resistant; FRT, flippase recognition target; Flp, flippase.