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. 2022 Dec 12;22:299. doi: 10.1186/s12866-022-02722-8

Table 2.

Primers used in this study

Name Sequence (5′–3′)
Primers for construction of mutants
 ∆rpoN-S-F GCTCTAGACAACCTGATGGCGGTACTG
 ∆rpoN-S-R CCTGTTGTCT GCAACCTCGGTATCCGTACT
 ∆rpoN-X-F CCGAGGTTGC AGACAACAGGATGAGGAAGACG
 ∆rpoN-X-R GGGGTACCCTGCCCATTTTTTCGCG
 ∆rpoN-F CAACCTGATGGCGGTACTG
 ∆rpoN-R CTGCCCATTTTTTCGCG
 ∆T2SS-2-S-F GCTCTAGAGTTAGTGAGTCGTCTATGTTGGCTT
 ∆T2SS-2-S-R GCTACGCCATCGTCACCCCGCCGTTAT
 ∆T2SS-2-X-F CGGGGTGACGATGGCGTAGCCGGTGGT
 ∆T2SS-2-X-R GGGGTACCCACCACGCCGCCTTGT
 ∆T2SS-2-F GTTAGTGAGTCGTCTATGTTGGCTT
 ∆T2SS-2-R CACCACGCCGCCTTGT
Primers for identification of plasmid
 pRE112-U-F CACTGTTCGTCCATTTCCG
 pRE112-D-R TTCGTCTCAGCCAATCCCT
 pBAD33-U-F AACAAAGCGGGACCAAAG
 pBAD33-D-R AGAGCGTTCACCGACAAA
 pMS402-U-F GGTCAAATGAATGCAGGGCT
 pMS402-D-R AGAGTCATTCAATATTGGCAGGT
Primers for construction of complemented strain
 pBAD33-rpoN+-F GGGGTACCATGAAGCCAAGTTTACAACTCAAG
 pBAD33-rpoN+-R GCTCTAGATTAAACCAAGCGTTTACGCTG
Primers for lux
 lux-flaC-F CCGCTCGAGATTTCTTGACATGCCGCGTT
 lux-flaC-R CGGGATCCATCTCCGTTAAACTTGCCGC
 lux-motA-F CCGCTCGAGTCCCCAGGTCTCAAAATCGT
 lux-motA-R CGGGATCCCATCAAACCTCTGTGCTCGT
 lux-fliAL-F CCGCTCGAGGCAAGCTGGCATCTCTGTAC
 lux-fliAL-R CGGGATCCCTATCCTCTGTCTACCGCGC
Primers for qRT-PCR
gyrB-RT-F GATTTGCGCACTGGGTAGCC
gyrB-RT-R GCGGCTGTTTGGATCCATGG
flaC-RT-F TTGCGATCGACTCATCCCTG
flaC-RT-R CAGGATCTGCTGCTTGGTCA
motA-RT-F GGTGGTTTTTTTCGCAGC
motA-RT-R TTCCTCTTACAAATAAAACGCC
fliAL-RT-F ATAGGTTTTGCCTTGCGATA
fliAL-RT-R TCTATCCTCTGTCTACCGCG
tssB-RT-F GTGGTCAGCAGGCCGAAATC
tssB-RT-R TTGGCACCCCTCTTCCAGAC
tssC-RT-F GGGTATTGCTGCGTTGGTGG
tssC-RT-R TGGTGGCATGCAAGACTTGG
tssE-RT-F TGTCATTTCAAGTCACAGCACA
tssE-RT-R GGTCACCTGCTCGATATTCTCCA
tssF-RT-F TTGTACGCGGCGATGAAACG
tssF-RT-R CAAATGACGGTGAGCTGCCC
tssG-RT-F CAATCGCGCCAAGATGCTGT
tssG-RT-R ACCTTATCGCCCAGCACCAA
tagH-RT-F ATCGTGAGCTGGCCAGTCAT
tagH-RT-R GGCTGGGATGCTGTAGGGAT
tssJ-RT-F ATGTGGCGATGAACCCGGAT
tssJ-RT-R CGTAATCGGTGGCCAACAGG
tssK-RT-F TGCTCGCTTAAGTGCCCTGA
tssK-RT-R GACCTCACTGACGCCACTGA
tssL-RT-F ACCATGCCGCACAGCTAGAT
tssL-RT-R TCTGTCACCACGCGCTGATA
tssH-RT-F CGAAAGCAAAGCAAACGCCG
tssH-RT-R CTTCGCCAACCACAATCGGG
vasH-RT-F ATTCACGACCGCCTGACTGT
vasH-RT-R GTGATCCAGTGTGCGCTTCG
tagO-RT-F AGGCTCCTGTGTCGGTTAGC
tagO-RT-R CCAAATGCGGGTAATGGCGT
tssA-RT-F TGCGCGAGATGACCTTCAGT
tssA-RT-R ACTCGCTGCCTAAATCCGCT
tssM-RT-F CAGTTTGAGTGCGGCGGATC
tssM-RT-R TCATTGCCAATCACCGCTGC
impA-RT-F CATTGCAGGGTATGGAGCGC
impA-RT-R CAAAGGCGAGGCTTGCTCAG
paaR-RT-F ATTACGGCCAGCCCTGATGT
paaR-RT-R GCTTATCGTGCGGCTCAAGG
vgrG1-RT-F GGGCATGCGGTGATTGTGTT
vgrG1-RT-R CCTGCCCAGCCTGATCTTCA
hcp-RT-F TCACCTCTGAATCCGTGGGC
hcp-RT-R GCGCAACGGTGAACTGGAAT
vgrG2-RT-F CCGGGTTGTGCGTTCAGTTT
vgrG2-RT-R CACGGTAGCGATTTGTGGGC
 orf00078-RT-F TACGCCTGAGTGATGCCCAG
 orf00078-RT-R TCCTCACTTCCCCACCATGC
 orf00080-RT-F TCGGCCAGGCGATACAACTT
 orf00080-RT-R CGGATCTCACGCCACTCAGT
yghF2-RT-F CAAGGATAACGTGCTGGTCG
yghF2-RT-R AATCCGGTAATTCACGCAGC
gspD2-RT-F CGCCAGATATCATGCAGTCG
gspD2-RT-R GCACGCCGAAATTGATGTTG
gspE2-RT-F GCCTTACTCACTAGCGCCTA
gspE2-RT-R TGGCGTTGATGAGCTTGATG
gspF2-RT-F TAGCCGAGCAGTGTGAGAAA
gspF2-RT-R TGGCGCAAAACAACTCATCA
gspG2-RT-F AGTGATGGTGGTGATCGTGA
gspG2-RT-R TCGGGTACACGCTGTTATCC
gspH2-RT-F CATTAGCTGGTCGACAAGCC
gspH2-RT-R TACTGTCGAGCCACACTTGT
gspI2-RT-F GTATTGGCGCAGTAAGTCGG
gspI2-RT-R TCATAGCGCTGACTCCGTAC
gspJ2-RT-F TCAGGTGGTTGATGGGGTAC
gspJ2-RT-R ATTGTAAAAGTCCGTCGCCG
gspK2-RT-F TTAGATGGGCAAGTCACCGT
gspK2-RT-R CGAACTGATGGCGTTGAACA
gspL2-RT-F TTTAGACGTGGCCCAGGATT
gspL2-RT-R ACATCGGGTTTTGTTTCGGG
gspM2-RT-F TGTTGGTTGGCGGATTGTAC
gspM2-RT-R CAATTGCCGTAACAGCTGCT
spaS-RT-F CAACCACACCAGTAGACGCA
spaS-RT-R AACGGATGTGAAATTGGCGC
spaR-RT-F TTCGCTGGATCAATCGTGCT
spaR-RT-R GTCATGCTGGGCCTGTTACT
spaQ-RT-F GTTTCACCATACCAGCCCGA
spaQ-RT-R ATGATTTGATGTTTGCGGGCAA
spaO-RT-F GCCAACCACGTGACAACATC
spaO-RT-R GTGACCGTTGATTTGCCACC
spaN-RT-F GCTGCTGATGTGGATTTCGC
spaN-RT-R CAGCCTTCAGATTCGGTGGT
invC-RT-F AGCTATCAAACACCGACGCA
invC-RT-R TTCGCGATCAGGGTAAACGG
invG-RT-F CGCCCATCCAGAGTGCTTAT
invG-RT-R GAGGATCTGGAGCAACTCGG
invF-RT-F ACAACGCGAACATTCACACG
invF-RT-R GCTGGGTGTTGGCAAATGAC
prgH-RT-F TGGCCGACAATTACCCAGAC
prgH-RT-R CGTTTGAGCTCCGTTTCTGC
prgI-RT-F CCGGCGTTGAGGACTTACAA
prgI-RT-R CGGAAGTTTTGTATGATGGCGG
sipA-RT-F TTGACTCGACATTCTGCGCT
 sipA-RT-R CAAATCCGGCCACAAAGCTC
sipD-RT-F GCCTGCCACGCTTGAAATTT
sipD-RT-R TCAGTGATCGGAAGGTTGCC
 sipC-RT-F GCCTCAAGCGCAGACTTACT
sipC-RT-R CTGGGGCAATTGATGGTCCT
sipB-RT-F GCCTCAAGCGCAGACTTACT
sipB-RT-R CTGGGGCAATTGATGGTCCT
spaT-RT-F GCTTCCGCTTCATCGAGTCT
spaT-RT-R GAAATGGTCTGGGAAGCGGT
spaP-RT-F TGCTTTACCACCGGCATCAT
spaP-RT-R GCGTCTGGGACCTGCTTTAT
invB-RT-F CGAGCTCGCCGTCTTGAATA
invB-RT-R AGCATTCACATCAGCCAGCA
invA-RT-F AGTTAACCACGCGCACCATA
invA-RT-R TCAAGGGCCACTGGAAAAGG
invE-RT-F CCAACACCCGCTCAAAACTG
invE-RT-R CAGCGTTTCGTTCAGTCTGC
invH-RT-F ATGCAGCGAGGATGGAGTCA
invH-RT-R ATGCCCGCTCAAATTCGCTA
hilA-RT-F TACAAAATCGCATTGCCGCC
hilA-RT-R AGCTGCAGTACAATGACGCT
iagB-RT-F GCAACGTTATGGTTATAGCTGGG
iagB-RT-R ACGTTCCCAAATTTTGTTAGCA
prgJ-RT-F ATTACACGCCAAGACACCGAT
prgJ-RT-R TCGAGGCGAGCTAAGATTGC
prgK-RT-F AGTCGCCCAGATGTTTCCTG
prgK-RT-R ACGCGGACAAGTGAATGGAT
orgA-RT-F TCAACAACAAGCCGAGCAGA
orgA-RT-R CATGGGCTAGCAACTGTCGA

Underlined letters show Kpn1 、sacI、Xba1、BamhI or XhoI restriction site

F/R: The upstream and downstream primers

S/X-F/R: The upstream and downstream primers for the upstream and downstream gene fragments of rpoN or T2SS-2 in the O45 genome

U/D-F/R: Upstream and downstream sequencing primers of plasmid