TABLE 1.
Bacterial strains and plasmids
| Strain or plasmid | Description | Reference or source |
|---|---|---|
| Strains | ||
| P. aeruginosa | ||
| PA1244 | Wild-type strain, pilin group I | 44 |
| PA1244N3 | Interruption of sigma factor rpoN with tetracycline cassette, Tcr | 43 |
| PA103 | Wild-type strain, pilin group II | 34 |
| PA103 | Interruption of wzy gene with a gentamicin cassette (aacC1), GmrwzyPaO11::aacC1 | 13 |
| PA683 | Wild-type strain, pilin group III | This study |
| E. coli | ||
| XL-Gold | endA1 supE44 thi-1 recA1 gryA96 relA1 lacHte | Stratagene |
| HB101 | supE44 hsdS20 recA13 ara-14 proA2 lacY1 galK2 rpsL20 xyl-5 mtl-1 | 4 |
| Plasmids | ||
| pHK734 | pTrc99a with E. coli phoA | 31 |
| pCR2.1-TOPO | 3.9-kb cloning vector, Apr | InVitrogen |
| pECAP | pMMB66EH with pHK734 phoA | This study |
| pECAPaas | pMMB66EH with mutated E. coli phoA that had an AAS stop extension added to the C-terminal lysine residue | This study |
| pECAPpep | pECAP with DNA coding for C-terminal TAWKPNYAPANAPKS stop | This study |
| pMMB66EH | 8,807-bp broad-host-range expression vector, IPTG inducible tac promoter, Ampr Cbr | 20 |
| pUCP26PilO | pUCP26 with pilO under the control of tac promoter, Tcr | 27 |
| pPAC24 | pMMB66EH with 1244 pilA | 6 |
| pPAC46 | pMMB66EH with 1244 pilAO | 6 |
| pUC46 | pilAO in pUC19 | 9 |
| pW136A | pPAC46 with pilA codon for W136 converted to A | This study |
| pK137A | pPAC46 with pilA codon for K137 converted to A | This study |
| pN139A | pPAC46 with pilA codon for N139 converted to A | This study |
| pY140A | pPAC46 with pilA codon for Y140 converted to A | This study |
| pP142A | pPAC46 with pilA codon for P142 converted to A | This study |
| pN144A | pPAC46 with pilA codon for N144 converted to A | This study |
| pP146A | pPAC46 with pilA codon for P146 converted to A | This study |
| pK147A | pPAC46 with pilA codon for K147 converted to A | This study |
| pS148A | pPAC46 with pilA codon for S148 converted to A | 9 |
| pRMCD28 | Apr, PhoA fusion vector | 12 |
| pPilAPhoA | pRMCD28 with 1244 pilA | This study |
| pPilAP | pMMB66EH with 1244 pilA-E. coli phoA fusion | This study |
| pPilAPaas | pPilAP with DNA coding for C-terminal AAS stop | This study |
| pPilAPpep | pPilAP with DNA coding for C-terminal TAWKPNYAPANAPKS stop | This study |
| pSAD100 | pUC18 with PA103 pilA | 27 |
| pSD5 | pMMB66EH with PA103 pilA | 27 |
| pMAM | pSD5 with DNA coding for C-terminal TAWKPNYAPANAPKS stop | This study |
| pPAL100 | pMal-cRI containing DNA coding for a malE-pilO gene fusion | 42 |
| pUCP24 | Broad-host-range cloning vector, pUC18 derived, Gmr | 50 |
| pUCP24PilO | pUCP24 with pilO under control of tac promoter | This study |
| p683PilA | pMMB66EH with PA683 pilA | This study |
| p683aas | p683pilA with DNA coding for C-terminal AAS stop | This study |
| p683pep | p683pilA with DNA coding for C-terminal TAWKPNYAPANAPKS stop | This study |