TABLE 4.
Strains and plasmids used in this study
| Strain | Genotype or descriptiona | Source or reference |
|---|---|---|
| Strains | ||
| E. coli | ||
| Top10 | Routine cloning host | Lab stock |
| DH5α | Routine cloning host | Lab stock |
| E. faecalis | ||
| OG1 | Wild-type laboratory strain isolated from oral sample (MLST 1) | 39 |
| SB35 | OG1 Δ(croR croS)3 | 26 |
| E. faecium | ||
| 1,141,733 | Wild-type reference strain, isolated from blood culture of hospitalized patient (MLST 327) | 40 |
| JL537 | 1,141,733 Δ(croR croS)4 | This study |
| 1,231,408 | Wild-type reference strain, isolated from blood culture of hospitalized patient (MLST 582) | 40 |
| Plasmids | ||
| pJH123 | Expression vector, constitutive P23S promoter (Cmr) | This study |
| pSLK252 | pbp5 from E. faecium 1,141,733 in pJH123 | This study |
| pSLK253 | pbp5 from E. faecium 1,231,408 in pJH123; carries M485A and Ser466 insertion | This study |
| pBK2 | cCF10-inducible expression vector with lacZ in MCS (Cmr) | 34 |
| pJLL105 | pBK2 with lacZ removed (Cmr) | This study |
| pJLL160 | croRS from E. faecium 1,141,733 in pBK2 (replaces lacZ) | This study |
| pJH086 | E. faecalis allelic-exchange vector (Cmr, repA V71G, lacZ pheS*) | This study |
| pJLL150 | Δ(croREfm croSEfm)4 deletion allele in pJH086 | This study |
a
Cmr, chloramphenicol resistance.