TABLE 1.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Descriptiona | Source or reference |
|---|---|---|
| Bacterial strains | ||
| E. faecium | ||
| 1,231,410 | Clade A skin and soft tissue infection isolate; VanA-type VRE; DAPs | 25, 26 |
| 1,141,733 | Clade B clinical isolate; VANs | 25, 26 |
| Population A | In vitro-evolved population with reduced CHX susceptibility from expt A, stocked from day 22 | This study |
| Population B | In vitro-evolved population with reduced CHX susceptibility from expt B, stocked from day 22 | This study |
| 410-P1 | E. faecium 410 in vitro-evolved population in BHI from day 21, trial 1 | This study |
| 410-P2 | E. faecium 410 in vitro-evolved population in BHI from day 21, trial 2 | This study |
| DAP-A1 | Mutants with reduced DAP susceptibility from population A | This study |
| DAP-A2 | Mutants with reduced DAP susceptibility from population A | This study |
| DAP-B1 | Mutants with reduced DAP susceptibility from population B | This study |
| DAP-B2 | Mutants with reduced DAP susceptibility from population B | This study |
| PB301 | E. faecium 410 EFTG_02287-88 deletion mutant | This study |
| PB302 | E. faecium DAP-A2 EFTG_02287-88 deletion mutant | This study |
| AH101 | PB301 transformed with empty pLZ12 plasmid | This study |
| AH102 | PB301 transformed with pAH201 | This study |
| PB303 | PB301 transformed with pPB202 | This study |
| E. coli EC1000 | E. coli cloning host; provides repA in trans; F− araD139(ara ABC-leu)7679 galU galK lacX74 rspL thi; repA of pWV01 in glgB; KANr | 74 |
| Plasmids | ||
| pLZ12 | E. coli and Streptococcus shuttle vector | 69 |
| pHA101 | Markerless counterselectable exchange plasmid | 24 |
| pAH201 | pLZ12 containing a 4.186-kb BamHI/BamHI-digested fragment with EFTG_02287-88 and predicted promoter | This study |
| pPB202 | pLZ12 containing a 4.186-kb BamHI/BamHI-digested fragment with EFTG_02287-88 and predicted promoter from strain DAP-A2 | This study |
| pPB301 | pHA101 containing a 1.998-kb EcoRI/BamHI-digested fragment with upstream and downstream regions flanking EFTG_02287-88 | This study |
a
VANs, vancomycin susceptible; DAPs, daptomycin susceptible; Km, kanamycin.