TABLE 1.
Bacterial strains used in this study
| Strain or plasmid | Genotype or description | Reference or source |
|---|---|---|
| Strains | ||
| Escherichia coli DH5α | F− φ80lacZΔM15 Δ(lacZYA-argF)U169recA1endA1hsdR17(rK− mK+) phoAsupE44thi-1gyrA96relA1 λ− | Invitrogen |
| Brucella abortus | ||
| 2308 | Virulent challenge strain | Laboratory stock |
| KH16 | 2308 ahpCD::cat; Cmr | This study |
| KH40 | 2308 ahpCD::bla; Apr | This study |
| KH2 | 2308 katE::cat; Cmr | This study |
| MEK6 | 2308 katE::aph3A; Knr | This study |
| KK9 | 2308 ahpCD::blakatE::aph3A; Apr Knr | This study |
| KK21 | 2308 ahpCD::catkatE::aph3A; Cmr Knr | This study |
| Plasmids | ||
| pGEM-T Easy | ColE1-based cloning vector; Apr | Promega |
| pBC KS+ | ColE1-based cloning vector; Cmr | Stratagene |
| pBBR1MCS-4 | pBBR-based broad-host-range cloning vector; moderate copy no. (10-14 copies per cell); Apr | 35 |
| pMR10-Ap | RK2-based broad-host-range cloning vector; low copy no. (2-4 copies per cell); Apr | 69 |
| pMR10 | RK2-based broad-host-range cloning vector; low copy no. (2-4 copies per cell); Knr | 22 |
| pBlue-CM2 | 656-bp cat gene from pBC cloned into the EcoRV site of pBluescript KS+ | 54 |
| pKS + Kan | 794-bp aph3A gene from TnphoA cloned into SalI-HindIII-digested pBluescript II KS+ | 35 |
| pMWV19 | 2,072-bp genomic DNA fragment from B. abortus 2308 containing ahpCD (PCR primers ahpCD-3F/ahpCD-3R) cloned into pGEM-T Easy | This study |
| pMEK21 | Derivative of pBBR1MCS-4 carrying the katE gene from B. abortus S19 | 21 |
| pMWV77 | 2,094-bp NotI fragment from pMWV19 containing ahpCD cloned into pMR10-Ap | This study |
| pKHS6 | 1,382-bp genomic DNA fragment from B. abortus 2308 containing ahpCD (PCR primers ahpCD-2F/ahpCD-2R) cloned into pBBR1MCS-4 | This study |
| pKHS2 | 2,499-bp genomic DNA fragment from B. abortus 2308 containing ahpCD (PCR primers ahpCD-1F/ahpCD-1R) cloned into pGEM-T Easy | This study |
| pKHS3 | Derivative of pKHS2 in which a 673-bp BsmBI/HindIII fragment internal to the ahpC and ahpD coding regions was replaced with the cat gene from pBlue-CM2 | This study |
| pKHS4 | 2,534-bp fragment from pKHS2 containing ahpCD cloned into pBC KS+ | This study |
| pKHS5 | Derivative of pKHS4 in which a 950-bp HindIII/HincII fragment internal to the ahpC and ahpD coding regions was replaced with the bla gene from pGEM-T Easy | This study |
| pKHS4 | 2,534-bp fragment from pKHS2 containing ahpCD cloned into pBC KS+ | This study |
| pMEK7-9 | 1,917-bp genomic DNA fragment from B. abortus S19 containing katE cloned into the PvuII site of pUC18 | 21 |
| pMEK7-9c | Derivative of pMEK7-9 in which a 1-kb PflMI/EcoRI fragment internal to the katE coding region was replaced with the cat gene from pBlue-CM2 | This study |
| pMEK7-9k | Derivative of pMEK7-9 in which a 1-kb PflMI/EcoRI fragment internal to the katE coding region was replaced with the aph3A gene from pKS + Kan | 21 |