TABLE 1.
B. pseudomallei strains and plasmids used in this study
| Strain or plasmid | Relevant genotype or propertiesa | Reference or source |
|---|---|---|
| B. pseudomallei strains | ||
| 1026b | Wild-type strain; clinical isolate | 12 |
| Bp50 | 1026b with Δ(amrRAB-oprA)::FRT | 7 |
| Bp58 | Bp50 with ΔbpeR::FRT | This study |
| Bp173 | Bp58 with mini-Tn7T | This study |
| Bp174 | Bp58 with mini-Tn7T-bpeR+ | This study |
| Bp175 | Bp58 with mini-Tn7T-P1-bpeR+ | This study |
| Bp207 | Bp50 with Δ(bpeAB-oprB)::FRT | This study |
| Bp216 | Bp207 with mini-Tn7T | This study |
| Bp227 | 1026b with Δ(bpeAB-oprB)::FRT | This study |
| Bp250 | Bp207 with mini-Tn7T-bpeA+B+-oprB+ | This study |
| Bp340 | 1026b with Δ(amrRAB-oprA) | This study |
| Bp400 | Bp227 with Δ(amrRAB-oprA) | This study |
| Plasmids | ||
| p34E-Tp1 | Apr Tpr; source of P1 promoter | 13 |
| pEXKm5 | Kmr; allele replacement vector optimized for Burkholderia spp. | 22 |
| pWSK29 | Apr; low-copy-number cloning vector | 41 |
| pUC18T-mini-Tn7T-Km-FRT | Apr Kmr; mobilizable mini-Tn7 base vector | 7 |
| pUC18T-mini-Tn7T-Zeo-FRT | Apr Zeor; mobilizable mini-Tn7 base vector | 7 |
| pFKM2 | Apr Kmr; source of FRT-nptII-FRT cassette | 7 |
| pFLPe2 | Zeor; curable Flp recombinase expression vector | 7 |
| pFLPe4 | Kmr; curable Flp recombinase expression vector | 7 |
| pFZE1 | Apr Zeor; source of FRT-PEM7-ble-FRT cassette | 7 |
| pPS856 | Apr Gmr; source of FRT-aacC1-FRT cassette | 15 |
| pPS1615 | Cmr; fosmid clone with a 35.5-kb 1026b DNA chromosomal insert containing the bpeR-bpeAB-oprB region | M. Jacobs, University of Washington |
| pPS2009 | Apr; pWSK29 with EcoRI fragment from pPS1615 containing the bpeR-bpeAB-oprB region | This study |
| pPS2254 | Apr Zeor; pGEM-T Easy (Promega, Madison, WI) with Δ(amrRAB-oprA)::FRT-PEM7-ble-FRT allele | This study |
| pPS2296 | Apr Zeor; pGEM-T Easy with ΔbpeR::FRT-PEM7-ble-FRT recombinant DNA fragment | This study |
| pPS2321 | Apr; pGEM-T Easy with bpeR+ | This study |
| pPS2323 | Apr Kmr; pUC18T-mini-Tn7T-Km-FRT with bpeR+ | This study |
| pPS2333 | Apr; pGEM-T Easy with P1-bpeR+ | This study |
| pPS2339 | Apr Zeor; pUC18T-mini-Tn7T-Zeo-FRT with P1-bpeR+ | This study |
| pPS2411 | Apr Gmr; pGEM-T Easy with Δ(bpeAB-oprB)::FRT-aacC1-FRT recombinant DNA fragment | This study |
| pPS2412 | Apr Kmr; pGEM-T Easy with Δ(bpeAB-oprB)::FRT-nptII-FRT recombinant DNA fragment | This study |
| pPS2482 | Apr Zeor; pUC18T-mini-Tn7T-Zeo-FRT with bpeA+B+-oprB+ 5′ region | This study |
| pPS2502 | Apr Zeor; pUC18T-mini-Tn7T-Zeo-FRT with bpeA+B+-oprB+ | This study |
| pPS2551 | Apr; pPS2254 with Δ(amrRAB-oprA) and FRT-PEM7-ble-FRT deleted | This study |
| pPS2557 | Kmr; pEXKm5 with Δ(amrRAB-oprA)-containing fragment from pPS2551 | This study |
a
Abbreviations: Ap, ampicillin; Cm, chloramphenicol; Gm, gentamicin; Km, kanamycin; Zeo, zeocin. All chromosomal mini-Tn7 insertions used in this study are located at the glmS2-associated attachment site (7). PEM7 is a synthetic prokaryotic promoter.