Strains and plasmids
| Strain or plasmid | Relevant genotype, description, or sequence | Reference or source |
|---|---|---|
| E. coli strains | ||
| DH5α | l− f80dlacZDM15D(lacZYA-argF)U169 recA1 endA hsdR17(rK− mK−) supE44 thi-1 gyrA relA1 | Life Technologies |
| S17-1 (λpir) | thi pro hsdR- hsdM+ ΔrecA RP4-2::TcMu-Km::Tn7 | 55 |
| P. aeruginosa strains | ||
| PA14 | Wild type | |
| bifA strain | Unmarked in-frame deletion of the bifA gene | 30 |
| bifA pilY1 strain | Unmarked in-frame deletions of bifA and pilY1 | 29 |
| bifA pilA strain | Unmarked in-frame deletions of bifA and pilA | 29 |
| bifA fimU strain | Unmarked in-frame deletions of bifA and fimU | This study |
| bifA pilV strain | Unmarked in-frame deletions of bifA and pilV | This study |
| bifA pilW strain | Unmarked in-frame deletions of bifA and pilW | This study |
| bifA pilX strain | Unmarked in-frame deletions of bifA and pilX | This study |
| bifA pilE strain | Unmarked in-frame deletions of bifA and pilE | This study |
| bifA fimU-pilVWXY1Y2E strain | Unmarked deletions of bifA and the fimU-pilVWXY1Y2E operon | This study |
| bifA pilD strain | Unmarked in-frame deletions of bifA and pilD | This study |
| pilX+::His strain | PA14 with chromosomal insertion of a C-terminal His6 tag in the pilX gene | This study |
| pilX(G−1F)::His strain | PA14 with chromosomal insertion of a His6 tag and mutation of G−1 to F in the pilX gene | This study |
| pilXΔLP::His strain | PA14 with chromosomal deletion of the pilX leader peptide sequence and insertion of a His6 epitope | This study |
| bifA pilX+::His strain | bifA mutant with chromosomal insertion of a C-terminal His6 tag in the pilX gene | This study |
| bifA pilX(G−1F)::His strain | bifA mutant with chromosomal insertion of a His6 tag and mutation of G−1 to F in the pilX gene | This study |
| bifA pilXΔLP::His strain | bifA mutant with chromosomal deletion of the pilX leader peptide sequence and insertion of a His6 epitope | This study |
| pilD pilX+::His strain | pilD mutant with chromosomal insertion of a C-terminal His6 tag in the pilX gene | This study |
| pilD pilXΔLP::His strain | pilD mutant with chromosomal deletion of the pilX leader peptide sequence and insertion of a His6 epitope | This study |
| Plasmids | ||
| pMQ72 | Shuttle vector for cloning in yeast and for arabinose-inducible gene expression; Gmr | 52 |
| pMQ30 | Shuttle vector for yeast cloning and Gram-negative allelic replacement; Gmr | 52 |
| pMQ30-fimU | Plasmid for in-frame deletion of fimU gene; Gmr | This study |
| pMQ30-pilV | Plasmid for in-frame deletion of pilV gene; Gmr | This study |
| pMQ30-pilW | Plasmid for in-frame deletion of pilW gene; Gmr | This study |
| pMQ30-pilX | Plasmid for in-frame deletion of pilX gene; Gmr | This study |
| pMQ30-pilE | Plasmid for in-frame deletion of pilE gene; Gmr | This study |
| pMQ30-pilD | Plasmid for in-frame deletion of pilD gene; Gmr | This study |
| pMQ30-fimUpilVWXY1Y2E | Plasmid for in-frame deletion of the fimU-pilVWXY1Y2E operon; Gmr | This study |
| pMQ30-pilX+::His | Plasmid for chromosomal insertion of a His6 tag in the pilX gene | This study |
| pMQ30-pilX(G−1F)::His | Plasmid for chromosomal insertion of a His6 epitope and an amino acid substitution of G−1 to F in pilX | This study |
| pMQ30-pilXΔLP::His | Plasmid for chromosomal deletion of the pilX leader peptide sequence and insertion of a His6 epitope | |
| pPilX-His | His-tagged pilX gene in pMQ72; Gmr | This study |
| pPilX(G−1A)-His | His-tagged pilX gene with the G−1A substitution cloned in pMQ72; Gmr | This study |
| pPilX(G−1D)-His | His-tagged pilX with the G−1D substitution cloned in pMQ72; Gmr | This study |
| pPilX(G−1F)-His | His-tagged pilX gene with the G−1F substitution cloned in pMQ72; Gmr | This study |
| pPilW-His | His-tagged pilW gene in pMQ72; Gmr | This study |
| pPilW(G−1F)-His | His-tagged pilW gene with the G−1F substitution cloned in pMQ72; Gmr | This study |
| pPilY1-His | His-tagged pilY1 gene cloned in the pMQ80 expression vector | 29 |