TABLE 1.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Relevant characteristic(s)a | Reference or source |
|---|---|---|
| Strains | ||
| Escherichia coli | ||
| SM10 | Mobilizing strain, transfer genes of RP4 integrated in chromosome; Kmr Sms | 14 |
| SM10λpir | SM10 with a λ prophage carrying the gene encoding the π protein | 8 |
| DH5α | High-efficiency transformation | Bethesda Research Laboratories |
| SURE | High-efficiency transformation; deficient in homologous recombination; Kmr Tcr | Stratagene |
| TOP10 | General cloning and blue/white screening | Invitrogen |
| B. pseudomallei | ||
| 1026b | Clinical isolate; Smr Tcs Tps | 3 |
| DD303-1 | 1026b::pDD303-1; Tcr | This study |
| PBS-1 | 1026b::pPBS-1; Tcr | 3 |
| DD301-1 | 1026b::pDD301-1; Tcr | This study |
| DD314-1 | 1026b::pDD314-1; Tcr | This study |
| DD315-1 | 1026b::pDD315-1; Tcr | This study |
| DD316-1 | 1026b::pDD316-1; Tcr | This study |
| Plasmids | ||
| pOT182 | pSUP102(Gm)::Tn5-OT182; Cmr Gmr Apr Tcr | 7 |
| pUCP28T | Broad-host-range vector; IncP OriT; pRO1600 ori; Tpr | 13 |
| pSKM11 | Positive selection cloning vector; IncP OriT; ColE1 ori; Apr Tcs | 9 |
| pC21Bg | 18.5-kb BglII fragment from C21 obtained by self-cloning; Apr Tcr | This study |
| pC21E | 16.0-kb EcoRI fragment from C21 obtained by self-cloning; Apr Tcr | This study |
| pDD226Nh | 31.0-kb NheI fragment from DD226 obtained by self-cloning; Apr Tcr | This study |
| pDD232E | 8.4-kb EcoRI fragment from DD232 obtained by self-cloning; Apr Tcr | This study |
| pDD235E | 11.6-kb EcoRI fragment from DD235 obtained by self-cloning; Apr Tcr | This study |
| pIR2 | pUCP28T containing a 670-bp PCR product encompassing gspC; Tpr | This study |
| pDD303-1 | pSKM11 containing 750-bp XhoI-BamHI fragment internal to gspE; Apr Tcr | This study |
| pPBS-1 | pSKM11 containing 3′ end of fliC and downstream DNA; Apr Tcr | 3 |
| pDD301-1 | pSKM11 containing 650-bp BamHI fragment internal to orfB; Apr Tcr | This study |
| pDD314-1 | pSKM11 containing a 390-bp PCR product internal to orfC; Apr Tcr | This study |
| pDD315-1 | pSKM11 containing a 480-bp PCR product internal to gspN; Apr Tcr | This study |
| pDD316-1 | pSKM11 containing 330-bp XhoI fragment internal to orfD; Apr Tcr | This study |
a
ori, origin of replication.