TABLE 1.
Bacterial strains and plasmids used in this work
| Strain or plasmid | Relevant features | Source or reference |
|---|---|---|
| E. coli strains | ||
| DH5α | F′ supE4 lacU169 (φ80 lacZΔM15) hsdR17 recA1 endA1 gyrA96 thi-1 relA1 | Clontech |
| HB101 | supE4 hsdS20 recA13 ara-1 proA2 lacY1 galK2 rpsL20 xyl-5 mtl-1 | Clontech |
| MC1061 (λpir) | hsdR mcrB araD139 Δ(araABC-leu) 7679 ΔlacX74 galI galK rpsL thi lysogenized with λpir bacteriophage | Our laboratory |
| BL21 (λDE3) | F−ompT hsdSB (rB− rB+) dcm gal λ(DE3) | 38 |
| P. multocida strains | ||
| PM25 | Wild type | Our laboratory |
| PM1002 | Same as PM25, but Rifr Spcr | Our laboratory |
| PM1077 | Same as PM1002, but carrying pUA961 | This study |
| PM1078 | Same as PM1002, but hgbA::pUA967 | This study |
| PM1079 | Same as PM1002, but PM0298 defective and carrying pUA964 | This study |
| Plasmids | ||
| pGEM-T | PCR cloning vector, Apr | Promega |
| pRK2013 | rep (ColE1), Mob+ Tra+ Kmr | 5 |
| pHRP309 | Broad-host-range cloning vector for transcriptional lacZ fusion construction, Gmr Mob+ | 23 |
| pUA826 | Mob+, R6K replicon, Apr Strr Spcr | 2 |
| pET-22b | Overexpression plasmid, Apr | Novagen |
| pGH436 | Same as pBR322, but carrying the resl-tet-resl construction | 35 |
| pIVET5mut | R6K replicon containing tnpR and lacZY promotorless genes | 15 |
| pUA958 | Same as pUA826, but carrying the promotorless tnpR gene | This study |
| pUA959 | Same as pGH436, but carrying the Cm cassette | This study |
| pUA960 | Same as pUA958, but carrying the res1-Cm-res1 construction | This study |
| pUA961 | Same as pUA960, but carrying a fusion between the P. multocida hgbA 5′ end and the tnpR gene | This study |
| pUA962 | Same as pGEM-T, but carrying the P. multocida PM0298-PM0299-hgbA operon | This study |
| pUA963 | Same as pET-22b, but carrying a fusion between the P. multocida hgbA promoterless gene and the T7 promoter | This study |
| pUA964 | Same as pHRP309, but carrying P. multocida ORFs PM0298 and PM0299 | This study |
| pUA965 | Same as pUA826, but carrying a 169-bp internal fragment of P. multocida ORF PM0298 | This study |
| pUA966 | Same as pUA826, but carrying a 258-bp internal fragment of P. multocida ORF PM0299 | This study |
| pUA967 | Same as pUA826, but carrying a 832-bp internal fragment of P. multocida hgbA gene | This study |