Skip to main content
. 2016 Mar 24;11(3):e0152013. doi: 10.1371/journal.pone.0152013

Table 2. Oligonucleotide primers used in this study.

Primer name Purpose Primer sequence
CL-N Amplification from pTW1 to construct mutant strains ATAGTGTTGGTATGATGCG
ToJH1Hd Amplification from pTW1 to construct mutant strains GACAAGCTTAGAGTTGAATA
TJ2b JHTW1 (umuDAbΔ2–59) construction GTCCAGGCGATACAGCCCAA
TJ1 JHKW1 (umuDAbΔ2–83) construction CATCAGCCCTCCTAACACAA
umuD’For JHKW1 (umuDAbΔ2–83) construction GGTTTGCCATCACCTGCACA
CL0 JHDS1 (umuDAb A83Y) construction AGCCAACTAAAGTCATTCG
CL10 JHDS1 (umuDAb A83Y) construction TTTTCATCCGCCTAAAG
CLK JHDS1 (umuDAb A83Y) construction TAACGCATAGGTTTCAGATTG
a118c-a119c_g122c_For JHTW2 (umuDAb K40P R41P) construction CGAGTGCCAGAATCTCAGGTTGCTTTTATTCCGCCTTGGCTTTTAGATAAC
a118c-a119c_g122c_Rev JHTW2 (umuDAb K40P R41P) construction GTTATCTAAAAGCCAAGGCGGAATAAAAGCAACCTGAGATTCTGGCACTCG
DSumuDfor JHDT1 (umuDAb K40P R41P A83Y) construction TCTTATTGATTTTAATTCGGC
CL-4 JHDT1 (umuDAb K40P R41P A83Y) construction CCTGCTTATGCAATGACAG
ExtraUDRev JHDT1 (umuDAb K40P R41P A83Y) construction GCCTGGACTTTCAGTGC
To81Rev JHDT1 (umuDAb K40P R41P A83Y) construction CTGAACGTATTTGATTGAGC
g70a JHMP1 (umuDAb E24K) construction CTGGACGTAAGGCCAAATACCAAAAGCCAACTAAAGT
g70 JHMP1 (umuDAb E24K) construction ACTTTAGTTGGCTTTTGGTATTTGGCCTTACGTCCAG
SDMFirF JH100 promoter mutation construction TTATCGTGCGTCTCTCAACGTTTGTAACGA
SDMFirR JH100 promoter mutation construction TCGTTACAAACGTTGAGAGACGCACGATAA
SDMPromIR2For JH101 promoter mutation construction TGAATTTGTAACGATGAGCTAGCAGATTATTTTAACTTG
SDMPromIR2Rev JH101 promoter mutation construction CAAGTTAAAATAATCTGCTAGCTCATCGTTACAAATTCA
SDMFir1AFor JH102 promoter mutation construction ATTATCGTGCGTCTCTTTGAATTTGTAACGA
SDMFir1ARev JH102 promoter mutation construction TCGTTACAAATTCAAAGAGACGCACGATAAT
SDMPstPromFor JH103 promoter mutation construction CTTGAATCTGCAGCGATTTCAAGTTAGATT
SDMPstPromRev JH103 promoter mutation construction GAAATCGCTGCAGATTCAAGTTGACGCACGATAA
SDMMidEndFor JH104 promoter mutation construction AATAATCTAACTTGGCGCACTTACAAATCAAGT
SDMMidEndRev JH104 promoter mutation construction ACTTGAATTTGTAAGTGCGCCAAGTTAGATTATT
2731RTFor2 JH100-JH104 promoter mutation construction ACGATGGGCATGGATGAAGTGG
17TJ1analog 17978 umuDAbˊ construction CATAATCGCCTCCATTTCAC
17TJ1analogFor 17978 umuDAbˊ construction GGTTTCCCATCACCAGC
E.coliNHisumuDS E. coli umuD expression ACCCACGCGCATGTCGTAAAAAGCACCCAATTGTTTATCAAGCCTGC
E.coliNHisumuDAS E. coli umuD expression CTTGGTTAGTTAGTTATTATCAGCGCATCGCCTTAACGA
ddrR#RTFor RT-qPCR on ddrR ATACCGAACAAGCCGAGCAT
ddrR#2RTRev RT-qPCR on ddrR AGGCATGACTAAAGCCAGCA
umuDAb#RTFor RT-qPCR on umuDAb GGAGCATGTCGAGCAGAGTC
umuDAb#2RTRev RT-qPCR on umuDAb; also inJH100-JH104 promoter mutation construction TCACCTGCTTTGGCCGTAAT
0445RTFor RT-qPCR on gst (ACIAD0445) ACCTGTACTCACTGATGGCG
0445RTRev RT-qPCR on gst (ACIAD0445) ACAGACCTCGTTTCGGATCA
ADP0724RTFor RT-qPCR on nrdA ATGACCGTCGTCGTACTCAC
ADP0724RTRev RT-qPCR on nrdA GCTGTGCAAATTCTTCGCCA
16SrRNA#RTFor RT-qPCR reference primer CCACACTGGGACTGAGACAC
16SrRNA#2RTRev RT-qPCR reference primer AACCAGGTAAGCCTCCTCCT