TABLE 2.
Bacterial strains and plasmids used
| Strain and plasmid | Description | Reference | ||
|---|---|---|---|---|
| Strains | ||||
| F. nucleatum 23726 | Parental strain (wild-type strain) | From ATCC | ||
| F. nucleatum CTI-2 | Wild-type strain, isolated from human CRC samples | (27) | ||
| F. periodonticum 33693 | Wild-type strain | From ATCC | ||
| F. nucleatum CW2 | ∆ftsX; an isogenic derivative of CW1 | (25) | ||
| F. nucleatum BG01 | ∆radD; an isogenic derivative of 23726 | (39) | ||
| F. nucleatum ZP05 | WT strain 23726 containing pZP4C | This study | ||
| F. nucleatum ZP06 | WT strain 23726 containing pZP4C(radD) | This study | ||
| F. nucleatum ZP07 | WT strain 23726 containing pZP4C(bamA) | This study | ||
| F. nucleatum ZP07a | WT strain 23726 containing pZP4C(control) | This study | ||
| F. nucleatum ZP07b | WT strain 23726 containing pZP4C(bamA)-P-bamA′ | This study | ||
| F. nucleatum ZP07c | WT strain 23726 containing pZP4C(bamA)-P-skp-lpxD-glpQ | This study | ||
| F. nucleatum ZP07d | WT strain 23726 containing pZP4C(bamA)-P-bamA′-skp-lpxD-glpQ | This study | ||
| F. nucleatum ZP08 | WT strain 23726 containing pZP4C(ftsZ) | This study | ||
| F. nucleatum ZP09 | WT strain 33693 containing pZP4C(clpBFp) | This study | ||
| F. nucleatum ZP10 | WT strain 33693 containing pZP4C(tnaAFp) | This study | ||
| F. nucleatum ZP11 | WT strain CTI-2 containing pZP4C(tnaACTI-2) | This study | ||
| E. coli DH5α | Cloning host | From NEB | ||
| Plasmids | ||||
| pCWU6 | E. coli/Fusobacterium shuttle vector, chloramphenicol /thiamphenicol resistance; cmR/thiaR | (25) | ||
| pRECas1 | The fdx promoter incorporating the riboswitch element E controls Cas9 expression | (44) | ||
| pIA33 | Pxyl::dcas9-opt Pgdh::sgRNA-rfp catP | (67) | ||
| pZP4C | Pfdx::dcas9-opt PrpsJFP::sgRNA-ftsX catP | This study | ||
| pZP4C(radD) | Pfdx::dcas9-opt PrpsJFP::sgRNA-radD catP | This study | ||
| pZP4C-mCherry | Pfdx::dcas9-opt PrpsJFP::PrpsJcd::mCherry catP | This study | ||
| pZP4C(control) | Pfdx::dcas9-opt catP | This study | ||
| pZP4C(bamA) | Pfdx::dcas9-opt PrpsJFP::sgRNA-bamA catP | This study | ||
| pZP4C(bamA)-P-bamA′ | Pfdx::dcas9-opt PrpsJFP::sgRNA-bamA PbamA-bamA’ catP | This study | ||
| pZP4C(bamA)-P-skp-lpxD-glpQ | Pfdx::dcas9-opt PrpsJFP::sgRNA-bamA PbamA-skp-lpxD-glpQ catP | This study | ||
| pZP4C(bamA)-P-bamA′-skp-lpxD-glpQ | Pfdx::dcas9-opt PrpsJFP::sgRNA-bamA PbamA-bamA-skp-lpxD-glpQ catP | This study | ||
| pZP4C(ftsZ) | Pfdx::dcas9-opt PrpsJFP::sgRNA-ftsZ catP | This study | ||
| pZP4C(clpBFp) | Pfdx::dcas9-opt PrpsJFP::sgRNA-clpBFP catP | This study | ||
| pZP4C(tnaAFp) | Pfdx::dcas9-opt PrpsJFP::sgRNA-tnaAFP catP | This study | ||
| pZP4C(tnaACTI-2) | Pfdx::dcas9-opt PrpsJFP::sgRNA-tnaACTI-2 catP | This study | ||