Table 1.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Phenotype or genotypea | Reference or source |
|---|---|---|
| Strains | ||
| E. coli | ||
| DH5α F′IQ | F′ ϕ80dlacZΔM15 Δ(lacZYA-argF)U169 deoR recA1 endA1 hsdR17(rK+ mK+) phoA supE44 λ− thi-1 gyrA96 relA1/F′ proAB+ lacIqZΔM15 zzf::Tn5 (Kmr) | 57 |
| XL1-Blue MRF′ | Δ(mcrA)183 Δ(mcrCB-hsdSMR-mrr)173 endA1 supE44 thi-1 recA1 gyrA96 relA1 lac [F′ proAB lacIqZΔM15 Tn10 (Tcr)]c | 58 |
| S17.1(λpir) | λ lysogenic S17-1 derivative producing π protein for replication of plasmids carrying oriR6K, Nals | 59 |
| B. abortus | ||
| Wild-type 2308 | Virulent, field isolated, wild type, Nalr, erythritol resistant | Laboratory stock |
| ΔcypAB mutant | B. abortus 2308 unmarked double-deletion mutant in cypA and cypB genes | This study |
| ΔcypAB(pDK51) mutant | B. abortus 2308 ΔcypAB mutant with plasmid pDK51, Ampr | This study |
| ΔcypAB(pcypA) mutant | B. abortus 2308 ΔcypAB mutant with plasmid pcypB, Ampr | This study |
| ΔcypAB(pcypB) mutant | B. abortus 2308 ΔcypAB mutant with plasmid pcypA, Ampr | This study |
| ΔcypAB(pcypAB) mutant | B. abortus 2308 ΔcypAB mutant with plasmid pcypAB, Ampr | This study |
| ΔcypAB(pcypBR55A/F60A) mutant | B. abortus 2308 ΔcypAB mutant with plasmid pcypBR55A/F60A , Ampr | This study |
| Plasmids | ||
| pK18mobSacB | Mobilizable cloning vector, Kmr Sucs | 36 |
| pK18ΔcypAB | ΔcypAB deletion cloned into pK18mobSacB | This study |
| pDK51 | pBBR1MCS-4 broad-host-range cloning vector (Ampr), pLac | 52 |
| pcypA | 1.1-kb ApaI/PstI fragment containing the B. abortus 2308 cypA gene cloned into pDK51, Ampr | This study |
| pcypB | 0.96-kb ApaI/PstI fragment containing the B. abortus 2308 cypB gene cloned into pDK51, Ampr | This study |
| pcypAB | 1.7-kb ApaI/PstI fragment containing the B. abortus 2308 cypA and cypB genes cloned into pDK51, Ampr | This study |
| pcypBR55A/F60A | B. abortus cypBK55A/F60A gene cloned into pDK51, Ampr | This study |
a
Abbreviations: Ampr, ampicillin resistance; Nalr, nalidixic acid resistance; Tcr, tetracycline resistance; Kmr, kanamycin resistance.