TABLE 1.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Relevant characteristic(s)a | Reference or source |
|---|---|---|
| Strains | ||
| E. faecium | ||
| TX1330 | Fecal isolate from community volunteer; Amps, Chls, Erys, Gens, Kanr, Strs, Tets; Cn-Adh− | 8, 37 |
| TX2466 | Blood isolate; Eryi, Kanr, Gens, Vanr | This study |
| TX0082 | Endocarditis isolate; Ampr, Chls, Eryr, Kanr, Gens, Vanr; Cn-Adh+ | This study |
| TX2555 | Perineal wound isolate; Chls, Eryr, Kanr, Gens, Vanr; Cn-Adh+ | 37 |
| TX6051 | TX0082Δacm, acm deletion mutant of TX0082, Chlr; Cn-Adh− | This study |
| TX6054 | TX2555acm::pTEX6052, acm insertion disruption mutant of TX2555, Genr, Cn-Adh− | This study |
| E. faecalis | ||
| JH2Sm::Tn916 | JH2 harboring Tn916; Gens, Strr, Tetr | 6 |
| TX6053 | JH2Sm::Tn916(pTEX6052); Strr, Tetr, Genr | This study |
| OG1RF | Laboratory strain; Rifr, Fusr, Chls, Gens | 35 |
| E. casseliflavus | ||
| UC73 | aph(2″)-Id containing strain; Genr | 59 |
| E. coli | ||
| DH5α | E. coli host strain for routine cloning | Stratagene |
| TX5500 | DH5α(pTEX5500ts); Chlr, Genr | This study |
| TX5501 | DH5α(pTEX5501ts); Chlr, Genr | This study |
| TX6050 | DH5α(pTEX6050); Chlr, Genr | This study |
| TX6052 | DH5α(pTEX6052); Genr | This study |
| Plasmids | ||
| pAM401 | Shuttle plasmid; Chlr and, in E. coli, Tetr | 64 |
| pAT18 | Shuttle plasmid; Eryr | 60 |
| pTV1-OK | pWV01 replicon-based ts vector; Kanr, Eryr | 14 |
| pTV1-ts | pE194 replicon-based ts vector; Chlr, Eryr | 66 |
| pAM401ts | pIP501 replicon-based ts vector; Chlr | 60 |
| pFW14 | Streptococcal integration vector; Chlr | 42 |
| pHS1 | E. faecalis mobilizable ts plasmid containing oriTTn916; Kanr, Genr | 1 |
| pTEX5500ts | Shuttle plasmid, ts in gram-positive hosts; Chlr, Genr | This study |
| pTEX5501ts | Mobilizable ts vector containing oriTTn916; Chlr, Genr | This study |
| pTEX6050 | Plasmid for acm deletion with flanking regions of the acm binding domain cloned into pTEX5500ts; Chlr, Genr | This study |
| pTEX6052 | acm insertion construct, intragenic fragment of acm cloned into pTEX5501ts with deletion of cat; Genr, Chls | This study |
a
Chl, chloramphenicol; Ery, erythromycin; Fus, fusidic acid; Gen, gentamicin; Kan, kanamycin; Rif, rifampin; Tet, tetracycline; Str, streptomycin; Van, vancomycin; and ts, temperature sensitive. Superscript “s” designates sensitivity, “r” designates resistance, and “i” designates intermediate susceptibility (MIC of 2 to 4 μg/ml for Ery); for aminoglycosides “r” is defined for enterococci as MIC > 500 for Gen and > 2,000 for Kan. Cn-Adh+, adherence to collagen types I and IV; Cn-Adh−, no adherence to collagen types I and IV.