TABLE 1.
Bacterial strains and plasmids
| Plasmid or strain | Relevant genotype or features | Source or reference |
|---|---|---|
| Strains | ||
| E. coli | ||
| DH5α Max Efficiency | F− ϕ80dlacZΔM15 Δ(lacZYA-argF)U169 recA1 endA1 hsdR17 (rK− mK+) phoA supE44 λ− thi-1 gyrA96 relA1 | Invitrogen |
| MC101 | HB101/pRK24 | B. Dupuy |
| MC135 | HB101 containing pRK24 and pMC123 | 48 |
| MC253 | HB101 containing pRK24 and pMC199 | This study |
| MC263 | HB101 containing pRK24 and pMC213 | This study |
| MC274 | HB101 containing pRK24 and pMC217 | This study |
| MC277 | HB101 containing pRK24 and pMC211 | This study |
| MC295 | HB101 containing pRK24 and pMC230 | This study |
| MC306 | HB101 containing pRK24 and pJS107::spo0A178::TargeTron | 69 |
| MC329 | HB101 containing pRK24 and pMC238 | This study |
| MC330 | HB101 containing pRK24 and pMC240 | This study |
| C. difficile | ||
| 630 | Clinical isolate | 97 |
| 630Δerm | Erms derivative of strain 630 | N. Minton and 98 |
| R20291 | Clinical isolate | 41 |
| MC282 | 630Δerm pMC211 | This study |
| MC283 | 630Δerm pMC217 | This study |
| MC296 | 630Δerm oppB::ermB | This study |
| MC301 | 630Δerm appA::ermB | This study |
| MC307 | 630Δerm oppB::ermB appA::ermB | This study |
| MC310 | 630Δerm spo0A::ermB | This study |
| MC322 | 630Δerm oppB::ermB/pMC213 | This study |
| MC323 | 630Δerm oppB::ermB appA::ermB/pMC213 | This study |
| MC324 | 630Δerm/pMC123 | This study |
| MC331 | 630Δerm appA::ermB/pMC238 | This study |
| MC332 | 630 Δerm oppB::ermB/pMC240 | This study |
| MC334 | 630Δerm oppB::ermB appA::ermB/pMC238 | This study |
| MC337 | 63 Δerm oppB::ermB/pMC123 | This study |
| MC338 | 630Δerm appA::ermB/pMC123 | This study |
| MC339 | 630Δerm oppB::ermB appA::ermB/pMC123 | This study |
| Plasmids | ||
| pRK24 | Tra+ Mob+ bla tet | 99 |
| pBL100 | Untargeted group II intron-TargeTron vector | 29 |
| pCR2.1 | bla kan | Invitrogen |
| pCE240 | C. difficile TargeTron construct based on pJIR750ai (group II intron, ermB::RAM ltrA) catP | C. Ellermeier (44) |
| pJS107-spo0A-178TT | Group II intron targeted to spo0A | 69 |
| pMC123 | E. coli-C. difficile shuttle vector, bla catP | 48 |
| pMC199 | pBL100 with group II intron targeted to oppB | This study |
| pMC211 | pMC123 with cprA promoter | This study |
| pMC213 | pMC123 with PoppBCAD operon | This study |
| pMC217 | pMC211 with CD0852 | This study |
| pMC220 | pCR2.1 with group II intron targeted to appA | This study |
| pMC226 | pCE240 with appA-targeted intron | This study |
| pMC230 | pMC123 with appA-targeted intron, ermB::RAM ltrA catP | This study |
| pMC237 | pCR2.1 with PappFDABC operon | This study |
| pMC238 | pMC123 with PappFDABC operon | This study |
| pMC239 | pCR2.1 with PoppBCADF from R20291 | This study |
| pMC240 | pMC123 with PoppBCADF from R20291 | This study |