TABLE 4.
Strains and plasmids used in this study
| Strains and plasmids | Characteristics | Source |
|---|---|---|
| Strains | ||
| C. difficile | ||
| R20291 | Hypervirulent PCR ribotype 027, isolated from an outbreak in 2004–2005 | Ref. 24 |
| R20291fliC::CT | fliC mutant derived from R20291 by ClosTron insertion | This study |
| GT1::CT | GT1 mutant derived from R20291 by ClosTron insertion | This study |
| GT2::CT | GT2 mutant derived from R20291 by ClosTron insertion | This study |
| GT3::CT | GT3 mutant derived from R20291 by ClosTron insertion | This study |
| CDR20291_0245::CT | CDR20291_0245 mutant derived from R20291 by ClosTron insertion | This study |
| CDR20291_0246::CT | CDR20291_0246 mutant derived from R20291 by ClosTron insertion | This study |
| R20291fliC:: CT::fliC | R20291fliC mutant derived from R20291 containing the pMTL-fliC plasmid | This study |
| GT1::CT::GT1 (GT1comp) | GT1 mutant derived from R20291 containing the pMTL-GT1 plasmid | This study |
| GT2::CT::GT2 (GT2comp) | GT2 mutant derived from R20291 containing the pMTL-GT2 plasmid | This study |
| GT3::CT::GT3 (GT3comp) | GT3 mutant derived from R20291 containing the pMTL-GT3 plasmid | This study |
| CDR20291_0245::CT::R0245 (CDR20291_0245comp) | ORF5 mutant derived from R20291 containing the pMTL-CDR20291_0245 plasmid | This study |
| CDR20291_0246::CT::R0246 (CDR202911_0246comp) | ORF6 mutant derived from R20291 containing the pMTL-CDR20291_0246 plasmid | This study |
| CD1426 | Hypervirulent PCR ribotype 023, isolated from an outbreak in 2010 | Queens Hospital Remford |
| CD1426_GT2::CT | GT2 mutant derived from CD1426 by ClosTron insertion | This study |
| CD1426_GT2::CT::GT2 (CD1426_GT2comp) | GT2 mutant derived from CD1426 containing the pMTL-GT2 plasmid | This study |
| 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 | Ref. 50 |
| Plasmids | ||
| pMLT007C-E2-R20291fliC | ClosTron plasmid retargeted to fliC at 344/345a | DNA 2.0 |
| pMLT007C-E2-GT1 | ClosTron plasmid retargeted to GT1 at 478/479s | DNA 2.0 |
| pMLT007C-E2-GT2 | ClosTron plasmid retargeted to GT2 at 1293/1294s | DNA 2.0 |
| pMLT007C-E2-GT3 | ClosTron plasmid retargeted to GT3 at 1800/1801s | DNA 2.0 |
| pMTL007C-E2-CDR20291_0245 | ClosTron plasmid retargeted to CDR20291_0245 at 147/148s | DNA 2.0 |
| pMTL007C-E2-CDR20291_0246 | ClosTron plasmid retargeted to CDR20291_0246 at 392/393a | DNA 2.0 |
| pMTL84153 | E. coli–C. difficile shuttle plasmid (pCD6;catP; ColE1+tra; fdx promoter) | Ref. 51 |
| pMTL-fliC | pMTL84151 with fliC cloned with its native promoter | This study |
| pMTL-GT1 | pMTL84153 with GT1 cloned behind the fdx promoter | This study |
| pMTL-GT2 | pMTL84153 with GT2 and GT3 cloned behind the fdx promoter | This study |
| pMTL-GT3 | pMTL84153 with GT3 cloned behind the fdx promoter | This study |
| pMTL-CDR20291_0245 | pMTL84153 with CDR20291_0245 cloned behind the fdx promoter | This study |
| pMTL-CDR20291_0246 | pMTL84153 with CDR20291_0246 cloned behind the fdx promoter | This study |