Table 2. Oligonucleotide primers.
Oligonucleotides | Sequence (5′ - 3′)a |
SacI_dsbA_1 | TTTGAGCTCCAAGAAGAGATCCCGATCGTCC |
EcoRI_dsbA_2 | TTTGAATTCCATGACTTTCTCCATTGGATTTATT |
EcoRI_dsbA_3 | AATGAATTCTAATCTCAACCCATGATTCGGTAT |
XbaI_dsbA_4 | TTTTCTAGAGATTAAACTGTTGCTGCCGTCAG |
SacI_dsbB_1 | TTTGAGCTCGTCTTCCTGCCAATGTT |
EcoRI_dsbB_2 | TTTGAATTCCACAGATAGATCCTTGTTAAAAAGA |
EcoRI_dsbB_3 | TTTGAATTCTAAGCCAATCGCATCGCTCAAT |
XbaI_dsabB_4 | TTTTCTAGATAGCATGGAGAGTGAGCCGCCACT |
EcoRI_dsbC_1 | ATTGAATTCGTGCAAACGTCTGGTGGT |
XbaI_dsbC_2 | ATTTCTAGAGAGCTCGTGACCCAGCAT |
SacI_toxRS_1 | TTTGAGCTCATTTGGAAATCACATCGCGCAAAC |
BamHI_toxRS_2 | TTTGGATCCTCCTAATCCGAACATCTAATGTCC |
BamHI_toxR_3 | TTTGGATCCAACCCTAACGATGCCATCAAAGT |
BamHI_toxRS_3 | TTTGGATCCTCCGATGACAATAGTGCAGAAAG |
XbaI_toxRS_4 | TTTTCTAGAATGACGTTTCCCCGCGGTGAG |
XbaI_lacZ_1 | TAATCTAGAACACATAACCCTGCAGTA |
XhoI_lacZ_2 | TTTCTCGAGCTCTACGGCGTACATCCCT |
XhoI_lacZ_3 | TTTCTCGAGTGCGTGTGGAATGTGACGAT |
SacI_lacZ_4 | ATGAGCTCTTATTGTGGGGATGACGCTTT |
SacI_dsbA_5′ | AATGAGCTCGCCACTTTACAAGAACCCCCG |
XbaI_dsbA_3′ | ATTTCTAGAGATTTACAAAGCCGATTAGCACTG |
SacI_dsbB_5′ | AATGAGCTCCAATTGAAACTGAAACTAATCCAAG |
XbaI_dsbB_3′ | AATTCTAGACTTTAAGCGCCTTTTTTATCAACC |
HindIII_toxR_5′_FLAG | AATAAGCTTATGTTCGGATTAGGACACAACTCA |
KpnI_toxR_3′_FLAG | AATGGTACCCTACTCACACACTTTGATGGCAT |
KpnI_toxRC293S_3′_FLAG | AATGGTACCCTACTCAGACACTTTGATGGCATCGTTA b |
toxRC236S_5′ | GGCTACCGTCAATCGAACTGAGCGTTAAAAAATACAATGA b |
toxRC236S_3′ | TCATTGTATTTTTTAACGCTCAGTTCGATTGACGGTAGCC b |
KpnI_toxRS_5′_FLAG | AATGGTACCCATGTTCGGATTAGGACACAACTCA |
BglII_toxRS_3′_FLAG | TTAAGATCTTTAAGAATTACTGAACAGTACGGT |
BamHI_ompU_5′ | ATTGGATCCTCCTAAATCGGGTCGGGTT |
BamHI_ompU_3′ | AATGGATCCGGCTCAGCCATTTTCGTGGC |
BamHI_toxT_5′ | TTAGGATCCGTATAGCAAAGCATATTCAGAGA |
BamHI_toxT_3′ | ATTGTCGACTAAATAAACGCAGAGAGCCATC |
c_FLAGtoxR_3′_F1 | TGTCATCGTCGTCCTTGTAGTCCATCTAATGTCCCAGTATCTCCCTGT |
c_FLAGtoxR_5′_F2 | GGGACAGGGAGATACTGGGACATTAGATGGACTACAAGGACGACGATGA |
c_FLAGtoxR_3′_F2 | CTACTCACACACTTTGATGGCAT |
c_FLAGtoxRC293S_3′_F2 | CTACTCAGACACTTTGATGGCAT b |
c_FLAGtoxRtoxS_3′_F2 | CTTTCTGCACTATTGTCATCGGTCTACTCACACACTTTGATGGCAT |
c_FLAGtoxR_5′_F3 | AACCAGTTAACGCTGAATTACATTC |
c_FLAGtoxRC293S_5′_F3 | GTTGCTAACCCTAACGATGCCATCAAAGTGTCTGAG b |
c_FLAGtoxRtoxS_5′_F3 | ATGCCATCAAAGTGTGTGAGTAGTCCGATGACAATAG |
rpoB_fwc | CTGTCTCAAGCCGGTTACAA |
rpoB_rvc | TTTCTACCAGTGCAGAGATGC |
VC0633_fw | CTCGCGTACGTCTAAACTTCTTGG |
VC0633_rv | CGGTTGTCTAGGCTGTTGTTAGAC |
VC0984_fw | CCTTCATCAGCCACTGTAGTGAAC |
VC0984_rv | GACCGCTATCAGAATAAGCAGTCG |
VC1854_fw | ACCCACTAGTGATCGATGAAGACG |
VC1854_rv | GCCATACTCAGCATATACACGAGC |
VCr001_fw | AGGGAGGAAGGTGGTTAAGT |
VCr001_rv | CGCTACACCTGAAATTCTACCC |
Restriction sites are underlined.
Bold letters indicate codons changed to obtain desired amino acid mutations.
Oligonucleotides for rpoB are according to reference [73].