TABLE 1.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Relevant characteristics | Source or reference |
|---|---|---|
| Acinetobacter strains | ||
| ADP1 | Wild type (strain BD413, ATCC 33305) | 26 |
| ADPU6 | PcaU binding site downstream of pcaU replaced by Kmr cassette | This study |
| E. coli strains | ||
| DH5α | General-purpose cloning strain | 20 |
| BL21(DE3) | F−ompT hsdSB(rB−mB−) gal dcm (DE3); gene for T7 RNA polymerase under control of lacUV5 integrated into the chromosome | 39 |
| Plasmids | ||
| pBend5 | Vector designed for circular permutation assay based on pBluescript SK(−) | 46 |
| pET-21(+) | Transcription vector with T7 promoter | 39 |
| pKOK6 | Contains promoterless lacZ-Kmr cassette | 28 |
| pZR15 | 7-kbp HpaII fragment containing pcaU inserted into pUC19 | 17 |
| pZR17 | 2,792-bp EcoRI fragment from pZR15 cloned into pUC19 | 17 |
| pAC7 | 2,792-bp EcoRI fragment from pZR15 cloned into pET-21(+), T7 promoter upstream of pcaU | This study |
| pZR18 | 2,618-bp HindIII-HpaII fragment containing pcaU inserted into pUC19 | 17 |
| pAC25 | Corresponds to pZR18 after removal of 218-bp SwaI-Eco47III fragment | This study |
| p5/128 | 128-bp EcoRI-BstUI fragment of the pcaU-pcaI intergenic region in HpaI site of pBend5 | This study |
| pAC29 | pZR17 with 130-bp BstBI fragment replaced by Kmr cassette from pKOK6 | This study |
| pAC48 | Contains pcaI′-pcaU, lacZ-Kmr cassette from pKOK6 fused to pcaI′ on vector pRK415 | P. Patz and U. Gerischer, unpublished results |
| pAC52 | 130-bp BstBI fragment deleted from pAC48 | This study |
| p5DR | Intergenic PcaU binding site without the external inverted 10-bp sequence repetition on vector pBend5 | This study |