Research Article

Molecular Characterization of Putative Virulence Determinants in Burkholderia pseudomallei

Table 2

Primers used for PCR and construction of mutants and complemented plasmids.

GenePrimer nameNucleotide sequence (5′ to 3′)Purpose

BPSL2033 2033IFAGAACTTCGAGCAATTGCTGInternal PCR
2033IRGAGAGATGACGTTCGGTCTTInternal PCR
M2033FGTGAACTGGTACAAAGAAATATCGMutant confirmation
M2033RCACGTTTCTCGGATAGAGCMutant confirmation
C2033FCTAGAGCGCGGCCTCGCGComplementation of BPSL2033::Km
C2033RCATCACTCGGCGCAATGAGACTGComplementation of BPSL2033::Km

BP1026B_I2784 2784IFGTCGAGAGTACGGTGTGTTCInternal PCR
2784IRCCTGCGAAATCCTTATCACInternal PCR
M2784FCTGTTTTCTAAGCGTCAGAAGMutant confirmation
M2784RAAATATGCAGGAAATAGCCCGMutant confirmation
C2784FCCTTCGCGCTGATTTGGTComplementation of BP1026B_I2784::Km
C2784RCTACTTCGTAGCTTGATGCGCCComplementation of BP1026B_I2784::Km

BP1026B_I2780 2780IFAAACCAGAAGGGCGATTTCInternal PCR
2780IRGCGTTCTTTTAAGAATTGGGTAGInternal PCR
M2780FCAGGTACGATTCATGGAACGMutant confirmation
M2780RGGTATTCCGTGACCTGAATGTMutant confirmation
C2780FAGGCTCGGAGTAGTAACACTTComplementation of BP1026B_I2780::Km
C2780RCTACTGCCTATGCTGGGGTATComplementation of BP1026B_I2780::Km

BURPS1106A_A0094 94IFAGTCGGGGGGTACACCTACInternal PCR
94IRCGACACCGAGGAAAATTTCInternal PCR
M94FGTGAATGTCGATCTTGCGMutant confirmation
M94RTCAATCTCCAGCGAGCTTMutant confirmation
C94FTTCTACCAGCGACTTGGCComplementation of BURPS1106A_A0094::Km
C94RTCAATCTCCAGCGAGCTTComplementation of BURPS1106A_A0094::Km

BURPS1106A_1131 1131IFAAGGAGTTGGGTACGTCGInternal PCR
1131RCCCTTGTGCCATTGATAGInternal PCR
pUT-RTTTGAGTGACACAGGAACACMutant confirmation
C1131FGTGGTTCAGCCAGGCACGComplementation of BURPS1106A_1131::Km
C1131RGTATGTGTCGTCCGCATTTGComplementation of BURPS1106A_1131::Km

BURPS1710A_1419 1419IFGCGATAGCGATTGGAAAACTInternal PCR
1419IRGAATCCAGACCCATTCCGTInternal PCR
KmFATGAGCCATATTCAACGGGAMutant confirmation
C1419FTGCACAAGCTGTTCAAATGComplementation of BURPS1710A_1419::Km
C1419RTTAAGGCTTGGGTGCAAGComplementation of BURPS1710A_1419::Km

BPSL3147 3147IFGACCAGTACGCGCTCAAGInternal PCR
3147IRGAACGAATACTTGTCGATCGInternal PCR
M3147FGATGTACACGTTCAACGACAAGMutant confirmation
M3147RCTCTTCCGGCATCTCGTAMutant confirmation
C3147FTTCAGGGTTACGAAGCGAAGComplementation of BPSL3147::Km
C3147RTCAGTGCAGCCGGATGCTCGComplementation of BPSL3147::Km