Table 1.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Relevant characteristic(s) and antibiotic resistance(s)a | Reference or source |
|---|---|---|
| Strains | ||
| E. faecium HM1070 | Wild-type parental strain; Fusr Rifr | 31 |
| E. faecium UCN90A | LSAPr mutant derived from HM1070 (selected in vitro with Lin) | This study |
| E. faecium UCN90B | LSAPr mutant derived from HM1070 (selected in vitro with Lin) | This study |
| E. faecium UCN91A | LSAPr mutant derived from HM1070 (selected in vitro with Da) | This study |
| E. faecium UCN91B | LSAPr mutant derived from HM1070 (selected in vitro with Da) | This study |
| E. faecium UCN92A | LSAPr mutant derived from HM1070 (selected in vitro with Tia) | This study |
| E. faecium UCN92B | LSAPr mutant derived from HM1070 (selected in vitro with Tia) | This study |
| E. faecium UCN94 | HM1070 derivative with allelic replacement of eat(A) by eat(A)v from UCN90B (LSAP phenotype) | This study |
| E. faecium UCN95 | UCN90B derivative with allelic replacement of eat(A)v by eat(A) from HM1070 (wild-type phenotype) | This study |
| E. faecium UCN80 | Susceptible strain (isolated from patient B) | 15 |
| E. faecium UCN80-1 | LSAPr mutant derived from UCN80 (selected in vivo with Q-D) | 15 |
| E. faecium UCN81 | Susceptible strain (isolated from patient C) | 15 |
| E. faecium UCN81-1 | LSAPr mutant derived from UCN81 (selected in vivo with Q-D) | 15 |
| E. faecium UCN82 | Susceptible strain (isolated from patient F) | 15 |
| E. faecium UCN82-1 | LSAPr mutant derived from UCN82 (selected in vivo with Q-D) | 15 |
| E. coli TOP10 | Cloning strain | Invitrogen |
| Plasmids | ||
| pCR2.1-TOPO | General AT cloning vector; Kanr Ampr | Invitrogen |
| pG1KT | Derivative of the thermosensitive shuttle plasmid pG(+)host5 (containing promoterless and terminatorless Kanr cassette AphA-3); used for mutagenesis; Eryr | 19 |
| pG1KTΩeat(A) | pG1KT derivative carrying the 3′ end of eat(A); used for allelic replacement in UCN90B | This study |
| pG1KTΩeat(A)v | pG1KT derivative carrying the 3′ end of eat(A)v; used for allelic replacement in HM1070 | This study |
a
Ampr, ampicillin resistance; Da, dalfopristin; Eryr, erythromycin resistance; Fusr, fusidic acid resistance; Kanr, kanamycin resistance; Lin, lincomycin; LSAP, lincosamides-streptogramins A-pleuromutilins (LSAP phenotype); Q-D, quinupristin-dalfopristin; Rifr, rifampin resistance; Tia, tiamulin; eat(A), wild-type allele; eat(A)v, allele of eat(A) with a single mutation (C1349T) leading to an amino acid substitution (Thr450Ile).