Table 2.
Bacterial Strains and plasmids
| Plasmid or Strain | Relevant genotype or features | Source, construction or reference |
|
|---|---|---|---|
| Strains | |||
| E. coli | |||
| HB101 | F− mcrB mrr hsdS20(rB− mB−) recA13 leuB6 ara-14 proA2 lacY1 galK2 xyl-5 mtl-1 rpsL20 | B. Dupuy | |
| MC101 | HB101 pRK24 | B. Dupuy | |
| DH5α Max Efficiency | F− Φ80lacZΔM15 Δ(lacZYA-argF) U169 recA1 endA1 hsdR17 (rk−, mk+) phoA supE44 λ−thi− 1 gyrA96 relA1 | Invitrogen | |
| MC341 | HB101 pRK24 pMC263 | This study | |
| MC344 | HB101 pRK24 pMC266 | This study | |
| C. difficile | |||
| R20291 | Clinical isolate; ribotype 027 | (Stabler et al., 2009) | |
| 630Δerm | ErmS derivative of strain 630 | N. Minton (Hussain et al., 2005) | |
| MC310 | 630Δerm spo0A::ermB | (Edwards et al., 2014) | |
| MC346 | 630Δerm eutG::ermB | This study | |
| MC394 | 630Δerm eutA::ermB | This study | |
| Plasmids | pCE240 | C. difficile Targetron construct based on pJIR750ai (group II intron, ermB::RAM ltrA) catP | C. Ellermeier (Ho and Ellermeier, 2011) |
| pRK24 | Tra+, Mob+; bla, tet | (Thomas and Smith, 1987) | |
| pCR2.1 | bla kan | Invitrogen | |
| pMC123 | E. coli-C. difficile shuttle vector; bla, catP | (McBride and Sonenshein, 2011b) | |
| pMC253 | pCR2.1 Group II intron targeting region for eutA | This study | |
| pMC256 | pCR2.1 Group II intron targeting region for eutG | This study | |
| pMC257 | pCE240 Group II intron targeted to eutA | This study | |
| pMC260 | pCE240 Group II intron targeted to eutG | This study | |
| pMC263 | pMC123 Group II intron targeted to eutA, ermB::RAM ltrA catP | This study | |
| pMC266 | pMC123 Group II intron targeted to eutG, ermB::RAM ltrA catP | This study | |