Table 1.
Strains and plasmids used in this study
| Strain or plasmid | Relevant description or genotypea | Source or reference |
|---|---|---|
| Strains | ||
| E. coli | ||
| Top10 | Routine cloning host | Invitrogen |
| BL21(DE3) | Protein overexpression host | Lab stock |
| E. faecalis | ||
| OG1RF | Wild-type reference strain (MLST 1) | 21 |
| CK119 | OG1RF ΔireK2 | 4 |
| CK121 | OG1RF ΔireP2 | 5 |
| CK164 | OG1RF ΔireB2 | This study |
| CK167 | OG1RF ΔireK2 ireB2 | This study |
| JG4 | Ceftriaxone-resistant suppressor mutant of CK119; carries D50A mutation in IreB | This study |
| T1 (SS498) | Wild-type (MLST 21), CDC reference strain | 22 |
| JL376 | T1 ΔireB2 | This study |
| Plasmids | ||
| pET28b | E. coli expression vector (Knr) | Novagen |
| pJLL20 | pET28b::ireB-His6 | This study |
| pJLL22 | pET28b::ireB T4A T7A-His6 | This study |
| pCLH32 | pET28b::ireB T4A-His6 | This study |
| pCLH33 | pET28b::ireB T7A-His6 | This study |
| pCJK111 | pET28b::ireK-n | 5 |
| pCJK112 | pET28b::ireP | 5 |
| pCJK142 | pET28b::ireK-n K41R | 5 |
| pJRG32 | E. faecalis allelic exchange vector with pheS* counterselectable marker (Emr) | 23 |
| pCJK174 | pJRG32::ΔireB2 (ΔE6-H84) | This study |
| pDL278p23 | E. faecalis expression vector, constitutive p23 promoter (Spr) | 24 |
| pJRG39 | pDL278p23::ireB-strep | This study |
| pJLL14 | pDL278p23::ireB T4A T7A-strep | This study |
| pCI3340 | E. coli/E. faecalis shuttle vector (Cmr) | 25 |
| pCJK187 | pCI3340::p23-ireB-strep | This study |
| pCLH140 | pCI3340::p23-ireB T4V-strep | This study |
| pCLH141 | pCI3340::p23-ireB T7V-strep | This study |
| pCLH142 | pCI3340::p23-ireB T4V T7V-strep | This study |
| pCLH111 | pCI3340::p23-ireB T4I-strep | This study |
| pCLH126 | pCI3340::p23-ireB T7I-strep | This study |
a
Cmr, chloramphenicol resistance; Emr, erythromycin resistance; Spr, spectinomycin resistance; Knr, kanamycin resistance.