Table 2 Oligonucleotides used for site directed mutagenesis, and overexpression
Oligonucleotide | Sequence | Description or reference |
|---|---|---|
pbrR C14S | 5’ CCA CCG GGG ATG CGG TGC 3’ | PbrR mutagenesis primer |
pbrR C55S | 5’ CCA GAG ACC GGG AGT GAC G 3’ | PbrR mutagenesis primer |
pbrR C79S | 5’ GAC TTC ACC GGA ATC CTG G 3’ | PbrR mutagenesis primer |
pbrR C114S | 5’ GGC ACC AGA AGA GGC TTC G 3’ | PbrR mutagenesis primer |
pbrR C123S | 5’ GCA GAA TCC CGG ACG ATT G 3’ | PbrR mutagenesis primer |
pbrR C132S | 5’ CGT ATC ACA CAC GGA GTC CGA C 3’ | PbrR mutagenesis primer |
pbrR C134S | 5’ CGT ATC AGA CAC GCA GTC CGA C 3’ | PbrR mutagenesis primer |
pbrR C132S/C134S | 5’ CGT ATC AGA CAC GGA GTC CGA C 3’ | PbrR mutagenesis primer |
pbrApe | 5’ GCG CCA ACC GTG CTC GGT TCT GGG 3’ | Primer extension/sequencing primer [4] |
pbrBstEII | 5’ GCG AAT GGT CAC CAC CGG 3’ | Primer to amplify PpbrA |
pbrNruI | 5’ GCT TGT CGC GAA TCA GCG 3’ | Primer to amplify PpbrA |
pMU forward | 5’ GAT TCT CCC CAC ATC ACC AG 3’ | Sequencing primer for pMU2385 |
pMU reverse | 5’ TGC CAG CAT TTC ATA ACC AA 3’ | Sequencing primer for pMU2385 |
M13-F | 5’ CGC CAG GGT TTT CCC AGT CAC GAC 3’ | Sequencing primer for pUK plasmids |
M13-R | 5’ GAG CGG ATA ACA ATT TCA CAC AGG 3’ | Sequencing primer for pUK plasmids |
conpbr: | 5’ CTAGAGGGTTAATCGGCAAC 3’ | PpbrA mutagenesis primer |
merpbr: | 5’ CTAGAGGGTGTAAGGTCGGCAAC 3’ | PpbrA mutagenesis primer |
-1EcoPbr | 5’ GGG GAA TTC GAA GCT TGC T 3’ (3’ primer) | PpbrA mutagenesis primer |
-1CentreBam | 5’ GCC GAT TTA AAC CCT CTA GT 3’ (primer B) | PpbrA mutagenesis primer |
-1CentreEco | 5’ CGG CTA AAT TTG GGA GAT CA 3’ (primer A) | PpbrA mutagenesis primer |
-1BamPbr | 5’ CAG TAT ACC TAG GCA GCT GG 3’ (5’ primer) | PpbrA mutagenesis primer |
pbrR ATG (LIC) | 5’ GAC GAC GAC AAG ATG AAT ATC CAG ATC GGC GAG C 3’ | PbrR cloning and overexpression primer |
pbrR TAG (LIC) | 5’ GAG GAG AAG CCC GGT CTA GTC GCT TGG ATG GGC 3’ | PbrR cloning and overexpression primer |
T7 terminator | 5’ CGA TCA ATA ACG AGT CGC C 3’ | Sequencing primer |