Table 2.
Bacterial strains and plasmids
| Strain/plasmid | Genotype or description | Reference(s) or source |
|---|---|---|
| Strains | ||
| E. coli | ||
| Top10 | F− mcrA Δ(mrr-hsdRMS-mcrBC) φ80lacZΔM15 ΔlacX74 nupG recA1 araD139 Δ(ara-leu)7697 galE15 galK16 rpsL(Strr) endA1 λ− | Invitrogen |
| CA434 | E. coli HB101 [F− mcrB mrr hsdS20(rB− mB−) recA13 leuB6 ara-14 proA2 lacY1 galK2 xyl-5 mtl-1 rpsL20(Smr) glnV44 λ−] containing plasmid R702 | Williams et al. (1990) |
| C. difficile | ||
| 630Δerm | Derivative of 630 strain, erythromycin sensitive | Hussain et al. (2005) |
| 630Δerm-fliC | fliC::CT insertion mutant derived from 630Δerm | Twine et al. (2009) |
| 630Δerm-0240 | 0240 mutant derived from 630Δerm by clostron insertion | Twine et al. (2009) |
| M68Δerm | M68Δerm: Derived from M68 by deletion of ermB | Faulds-Pain and Wren (2013) |
| M68ΔfliC | M68ΔfliC: Derived from M68 by deletion of fliC | Faulds-Pain and Wren (2013) |
| 630Δerm-0241::CT | 0241 mutant derived from 630Δerm by clostron insertion | This study |
| 630Δerm-0242::CT | 0242 mutant derived from 630Δerm by clostron insertion | This study |
| 630Δerm-0243::CT | 0243 mutant derived from 630Δerm by clostron insertion | This study |
| 630Δerm-0244::CT | 0244 mutant derived from 630Δerm by clostron insertion | This study |
| M68Δerm-0242::CT | 0242 mutant derived from M68Δerm by clostron insertion | This study |
| M68Δerm-0243::CT | 0243 mutant derived from M68Δerm by clostron insertion | This study |
| 630Δerm-0241 ΔfliC | Double mutant fliC deletion mutant derived from 630Δerm-0241::CT | This study |
| 630Δerm-0242 ΔfliC | Double mutant fliC deletion mutant derived from 630Δerm-0242::CT | This study |
| 630Δerm-0244 ΔfliC | Double mutant fliC deletion mutant derived from 630Δerm-0244::CT | This study |
| Plasmids | ||
| pMTL84151 | E. coli – C. difficile shuttle plasmid (pCD6;catP; ColE1+tra) | Heap et al. (2009) |
| pfliC_comp | pMTL84151, with C. difficile 630 fliC and its native promoter | This study |
| pMTL84153 | E. coli – C. difficile shuttle plasmid (pCD6;catP; ColE1+tra; fdx promoter) | Heap et al. (2009) |
| p0241_comp | pMTL84153 with CD0241 cloned behind the fdx promoter | This study |
| p0242_comp | pMTL84153 with CD0242 cloned behind the fdx promoter | This study |
| p0244_comp | pMTL84153 with CD0244 cloned behind the fdx promoter | This study |
| pMTL007C-E2-CD0241 | Clostron plasmid retargeted to CD0241 at 231/232s (DNA 2.0) | This study |
| pMTL007C-E2-CD0242 | Clostron plasmid retargeted to CD0242 at 156/157s (DNA 2.0) | This study |
| pMTL007C-E2-CD0243 | Clostron plasmid retargeted to CD0243 at 309/310a (DNA 2.0) | This study |
| pMTL007C-E2-CD0244 | Clostron plasmid retargeted to CD0244 at 556/557a (DNA 2.0) | This study |
| pMTLAFPfliC | pMTL82151 with + /− 630fliC 1200 bp | Faulds-Pain and Wren (2013) |