TABLE 1.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Description | Source or reference |
|---|---|---|
| B. avium strains | ||
| 4169 | Wild type | 34 |
| 4169rif | Spontaneous Rifr mutant of 4169 | This study |
| Pho20 | 4169rif bhuR::TnphoA | 5 |
| Pho20(fur) | Pho20, fur gene interrupted by insertional integration of ptr5-1 | This study |
| E. coli strains | ||
| H1717 | Chromosomal lacZ under the control of a strong Fur-regulated promoter | 40 |
| MGIQ1 | K-12 derivative; lacIq1 Kanr | V. J. Hernandez |
| AF633 | MC4100 λφ(uspB-lacZ) | 11 |
| Plasmids | ||
| pCOS5 | Cosmid vector; Ampr Tetr | 4 |
| p1016 | 45-kbp fragment of 4169 chromosome encoding the rhuI-rhuR-bhuR locus ligated into pCOS5 | Murphy and Connell, unpublished |
| pMECA | Cloning vector, Ampr | W. A. Parrott |
| pERM1 | 3-kbp EcoRI fragment of p1016 ligated into pMECA | Murphy and Connell, unpublished |
| pBluescript KS | High-copy-number expression vector; Ampr | Stratagene |
| pAEK10 | 199-bp rhuR-bhuR intergenic region ligated into pBluescript KS | This study |
| pAEK13 | 142-bp rhuI promoter region ligated into pBluescript KS | This study |
| p1910 | Mobilizable ColE1 replicon; Ampr | S. Stibitz |
| ptr5-1 | Truncated fur with the ATG start codon mutated to TGA; mutant gene ligated into p1910 | 29 |
| pMGJ66 | lacIq1-containing plasmid; Tetr | M. G. Jobling |
| placI | lacIq1 allelle from pMGJ66 ligated into pUFR047 | This study |
| pET21a | Expression vector; Ampr | Novagen |
| pFI-7 | rhuI ORF ligated into pET21a | This study |
| pRK415 | Bordetella spp. expression vector, Tetr | 19 |
| pERM26 | rhuI ORF from pFI-7 ligated into pRK415 | This study |
| pERM26.4 | rhuI frameshifted at the XhoI site ligated into pRK415 | This study |
| pRK415Δ | Alpha peptide gene in pRK415 frameshifted at the XhoI site | This study |
| pRS415 | Promoter reporter vector; Ampr | 38 |
| pDJM31 | rhuR-bhuR intergenic region ligated into pRS415 | This study |
| pUFR047 | E. coli/Bordetella spp. shuttle vector; Ampr Gmr | 6 |
| pDJM41 | Fragment of pDJM31 containing the tandem transcription terminators, the rhuR-bhuR intergenic region, and the lacZYA locus ligated into pUFR047 | This study |