Table 1. Strains, plasmids and primers used in this study.
| Strain or Plasmid | Relevant Propertiesa | Reference or Source |
| B. pseudomallei | ||
| 1026b | AG and ML resistant wild-type strain; clinical isolate | [45] |
| DD503 | AG and ML susceptible Δ(amrR-amrAB-oprA)1026b derivative | [13] |
| 708a | AG and ML susceptible clinical isolate | [16] |
| 2188a | AG and ML susceptible clinical isolate | [16] |
| 3799a | AG and ML susceptible clinical isolate | [16] |
| Bp24 | Spontaneous AG and ML resistant derivative of 3799a | This study |
| Bp35 | Spontaneous AG and ML resistant derivative of 2188a | This study |
| Bp50 | 1026b with Δ(amrR-amrAB-oprA) | [20] |
| Bp66 | Low level Gmr derivative of 708a | This study |
| Bp187 | Bp24 with Δ(amrR-amrRAB-oprA) | This study |
| Bp202 | Bp187::mini-Tn7T-LAC | This study |
| Bp194 | Bp187::mini-Tn7T-LAC-amrA + B +-oprA + | This study |
| Bp192 | Bp35 with Δ(amrR-amrAB-oprA) | This study |
| Bp201 | Bp192::mini-Tn7T-LAC | This study |
| Bp200 | Bp192::mini-Tn7T-LAC-amrA + B +-oprA + | This study |
| Plasmids | ||
| pEX-S12pheS | Gmr; gene replacement vector | Lopez and Schweizer, unpublished |
| pUC18T-mini-Tn7T-LAC | Apr, Gmr; mini-Tn7 cloning and delivery vector | [46] |
| pPS2142 | Apr, Gmr; pUC18T-miniTn7T-LAC with amrA+B+-oprA+; amrAB-oprA expression under Ptac b control | [20] |
| pTNS3 | Apr; source of Tn7 transposase components TnsABCD | [20] |
| pFKM2 | Apr Kmr; source of FRT-nptII-FRT cassette | [20] |
| pFLPe2b | Zeor; source of Flpe recombinase | [20] |
| pPS1927 | Apr; pWSK29 [47] with ∼15 kb strain 1026b chromosomal EcoRI fragment containing amrA+B+-oprA+ | This study |
| pPS2282 | Apr; pGEM-T Easy (Novagen) with ∼3.1 kb PCR fragment containing Δ(amrAB-OprA)::FRT-nptII-FRT t | This Study |
| pPS2354 | Gmr Kmr; pEX-S12pheS with ∼3.1 kb blunt-ended EcoRI fragment of pPS2282 cloned into the SmaI site | This Study |
| Primers c | ||
| 597 | 5′-CGAATTGGGGATCTTGAAGTTCCT | This study |
| 1546 | 5′-TACATGGCGATAGCTAGACTGG | This study |
| 1599 | 5′-CGCGCGCAATTGTTCCTC | This study |
| 1600 | 5′-TCGTAAGAAAGCGACACGCA | This study |
| 1601 | 5′-CGATTCTTCGCGCGTCTTG | This study |
| 1602 | 5′-CGCGTGCGTGCCCATTCG | This study |
| 1742 | 5′-AAGACCGCGCTCTATTACGA | This study |
| 1743 | 5′-TCGTCACCGTATCAGTGCAT | This study |
| 1756 | 5′-ATCTTGCCGTTGAAGTGTCC | This study |
| 1757 | 5′-ATCGCTGAACACGAAGAACC | This study |
| 1774 | 5′-ACTAGTAGTGAGCGCAACGCAATTA | This study |
| 1779 | 5′-GCCTCTTCGCTATTACGC | This study |
| 1797 | 5′-GTTCGTCGCCGAGGAGT | This study |
| 1801 | 5′-GAAGCCGGTGAAATCGACG | This study |
| 1954 | 5′-CTCAAGTCGGTGTCCATTCC | This study |
| 1955 | 5′-ACGTTATCCGGCGTGATCT | This study |
| 2031 | 5′-CCTGGTTCACCTGCTCGATG | This study |
| 2032 | 5′-CTTCGTCGCTGCAAGAAACG | This study |
| 2033 | 5′-CGATCGACCTGCCTGAAACC | This study |
| 2034 | 5′-AGCTCGTCGTGAACACGGC | This study |
| 2035 | 5′-GACGTAATGGAACGACGCGC | This study |
| 2036 | 5′-CGTCGGCGCATTGAACGACA | This study |
| 2037 | 5′-CGATTCGTACATCGCGGCGA | This study |
| 2038 | 5′-CTCAACTTCACGGGCGAGAT | This study |
a
Abbreviations: AG, aminoglycosides; Ap, ampicillin; Gm, gentamicin; Km, kanamycin; ML, macrolides; r, resistance; Zeo, zeocin.
b
Ptac, E. coli lac/trp operon hybrid promoter.
c
Only selected primers are shown; other primer sequences are available from the authors upon request. Oligonucleotides were purchased from IDT, Coralville, IA.