Table 1.

Strains and plasmids used in this study

Strain or plasmidDescriptionaReference
Strains
E. coliSM10λpirSM10 with a λ prophage carrying the gene encoding the π protein; Kmr 26
B. pseudomallei
  1026bClinical isolate; AGr Tcs 11
  C211026b derivative;gspD::Tn5-OT182 AGrTcs 10
  RM1011026b derivative; amrA::Tn5-OT182 AGs Tcr This study
  RM1021026b derivative; amrB::Tn5-OT182 AGs Tcr This study
  RM1041026b derivative; 1026b::pSK240 Tcr This study
  RM102 Smr RM102 derivative; rpsL(Smr)This study
  DD502RM102 Smr::pDD502 Sms KmrTcr This study
  DD503DD502 derivative; Δ(amrR-oprA) rpsL (Smr) AGs Tcs This study
Plasmids
 pOT182PSUP102(Gm)::Tn5-OT182 Cmr Gmr AprTcr 24
 pUCP28TBroad-host-range vector; IncP OriT; pRO1600 ori; Tpr 36
 p34ECassette cloning vector; Apr 42
 pKAS46Mobilizable allelic exchange vector; π-dependent R6K replicon; Apr Kmr rpsL +(Sms) 37
 pSKM11Mobilizable allelic exchange vector; TcrApr 27
 pSKM240Mobilizable allelic exchange vector; pSKM11 containing a 240-bp internalSst1 fragment from amrR; TcrApr This study
 pBluescript SKGeneral cloning vector; Apr Stratagene
 pAW2214.0-kbNotI fragment from RM101 obtained by self-cloning; Apr Tcr This study
 pAW141.0-kbSalI fragment from RM101 obtained by self-cloning; Apr Tcr This study
 pAW3112.1-kbBamHI fragment from RM102 obtained by self-cloning; Apr Tcr This study
 pAW3219.3-kbBamHI fragment from RM101 obtained by self-cloning; Apr Tcr This study
 pRM105pUCP28T containing a 14.7-kb EcoRI fragment from pAW32This study
 pSR20pSK-Bluescript containing a 4.0-kb SacI fragment from pAW32This study
 pDD500pKAS46 derivative; 2.4-kb SacI-EcoRI fragment from pSR20 containing 3′ end of oprA and 1.4 kb of downstream DNAThis study
 pDD501p34E derivative; 2.1-kb BamHI fragment from pAW22 containing the 3′ end of amrR and 1.5 kb of downstream DNAThis study
 pDD502pDD500 containing a 2.11-kb XbaI fragment from pDD501 (XbaI sites flank the BamHI sites in pDD501)This study
  • a r, resistant; s, susceptible; AG, aminoglycoside; Tc, tetracycline; Ap, ampicillin; Sm, streptomycin; Km, kanamycin.