TABLE 1.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Genotype or description | Reference or source |
|---|---|---|
| Strains | ||
| C. crescentus | ||
| NA1000 | syn-1000 | 16 |
| LS107 | syn-1000 bla-6 | Dickon Alley |
| LS800 | rec-526 (pKM3001) | 37 |
| LS1218 | syn-1000 ΔfliF | 22 |
| LS1917 | syn1000 flgI::Tn5 | 32 |
| LS2195 | syn-1000 ctrA401 | 39 |
| LS2821 | syn-1000 fliX::Tn5 | This work |
| LS2996 | rec526 fliX::Tn5 | This work |
| SC508 | flaS153 (ΔfliQR) | 24 |
| SC1117 | flbN194::Tn5 str-152 | 10 |
| SC1131 | fliLM196::Tn5 | 57 |
| E. coli | ||
| TG1 | F′ lacIqproA+B+ lacZΔM15 supE44 | 7 |
| S17-1 | Integrated RP4-2, Tc::Mu, Km::Tn7 | 46 |
| Plasmids | ||
| pCM1 | pLAFR5-derived cosmid containing flgI | 32 |
| pCM2 | 5.3-kb SacI fragment containing flgI subcloned from pCM1 into pSKII+ | 32 |
| pCM9 | 800-bp PstI fragment subcloned from pCM2 into pRKlac290 | This work |
| pCM10 | 800-bp PstI fragment subcloned from pCM2 into pRKlac290 | This work |
| pCM11 | 5.3-kb SacI fragment containing flgI and fliX subcloned from pCM2 into pMR4 | This work |
| pCM12 | 1.3-kb SacI/XhoI fragment containing fliX subcloned from pCM2 into pMR4 | This work |
| pCM13 | 600-bp NlaIII fragment containing fliX subcloned from pCM2 into pMR4 | This work |
| pCM14 | 400-bp PstI/NlaIII fragment containing truncated fliX subcloned from pCM2 into pMR4 | This work |
| pCM15 | 613-bp PstI/XhoI fragment containing the fliX-flgI intergenic region subcloned from pCM2 into pSKII(+) | This work |
| pCM16 | 613-bp PstI/XhoI fragment containing the fliX-flgI intergenic region subcloned from pCM2 into pSKII(−) | This work |
| pCM17 | fliX cloned as a 420-bp EcoRI/HindIII fragment (generated by PCR) into pET21b | This work |
| pCM18 | 1.1-kb PstI fragment containing dksA subcloned from pCM1 into pSKII(+) | This work |
| pRKlac290 | lacZ transcriptional fusion vector, Tetr IncP1 replicon, mob+ | 20 |
| pMR4 | Broad-host-range vector, Tetr | C. Mohr and R. Roberts |
| pSKII(+) | Cloning and single-stranded phagemid | Stratagene |
| pSKII(−) | Cloning and single-stranded phagemid | Stratagene |
| pET21b | Ampr vector with T7 promoter for protein overexpression and generation of carboxyl-terminal polyhistidine tag | Novagen |
| pKM3001 | Pseudomonas aeruginosa recA-containing fragment in the broad-host-range vector pCP13 (Tetr) | 27 |