The QseB/QseC two-component system contributes to virulence of Actinobacillus pleuropneumoniae by downregulating apf gene cluster transcription

Duan, Benzhen; Peng, Wei; Yan, Kang; Liu, Feng; Tang, Jia; Yang, Fengming; Chen, Huanchun; Yuan, Fangyan; Bei, Weicheng

doi:10.1186/s44149-022-00036-w

Table 2 Primers used in this study

From: The QseB/QseC two-component system contributes to virulence of Actinobacillus pleuropneumoniae by downregulating apf gene cluster transcription

Primers	Sequence (5′-3′)^a
For mutant construction
qseBC-up-F/R	TTGTCGACCTATGGCTTTAACTTCCTTCG
qseBC-up-F/R	GGGAATTCATTATTGTCCGATAAAGCGAC
qseBC-down-F/R	GGGAATTCTTAAGCCGAAGAGAAAAAGACG
qseBC-down-F/R	GGTCTAGAGTCGTATCGTTTAATGCGATC
apfA-up-F/R	CTGTCGACTTTGATACCGAGCTGCAATAAGCGT
apfA-up-F/R	GCGTTTACTTTAATTGAATTGATGAGGAAACGAAATGTAAAGGGGAGGAC
apfA-down-F/R	GTCCTCCCCTTTACATTTCGTTTCCTCATCAATTCAATTAAAGTAAACGC
apfA-down-F/R	ATGCGGCCGCCGCTACGGCTGAGGAAAATC
apfB-up-F/R	CTGTCGACCGATAATCAGCATTAAGATCGGCTC
apfB-up-F/R	GAAATACCTGCTCGGGCAATTAATATTGTTATCACGGCTACAGAGGC
apfB-down-F/R	GCCTCTGTAGCCGTGATAACAATATTAATTGCCCGAGCAGGTATTTC
apfB-down-F/R	ATGCGGCCGCTACATTCTTTGCATTGCCCCAT
apfC-up-F/R	CTGTCGACGGCTTGTATTTGGCTTAACATCTCT
apfC-up-F/R	ACAACTTCAGAAAGGAAAAGCGTATTATCGGTACGGTTTTGATCGGAT
apfC-down-F/R	ATCCGATCAAAACCGTACCGATAATACGCTTTTCCTTTCTGAAGTTGT
apfC-down-F/R	ATGCGGCCGCCGAGATTTTTGCTTTTCTTACGC
apfD-up-F/R	CTGTCGACCATTTGCTTTCCTTATCTTTTGCTA
apfD-up-F/R	GTGGTGTAAGCGAAGTATTCCTCAATTTTCGACTTTCGTGCTAATTATTG
apfD-down-F/R	CAATAATTAGCACGAAAGTCGAAAATTGAGGAATACTTCGCTTACACCAC
apfD-down-F/R	ATGCGGCCGCGCTGAATGATTTAAACCGTTTGATA
apxIVA-F/R	GGCTACCCATTTCCCTTCG
apxIVA-F/R	GAGCAACAACGTCGCACA
qseBC-exterior-F/R	GTTTATCGGTAGGTCCACAGG
qseBC-exterior-F/R	TGATCGCAGCAAATACCAAA
qseBC-interior-F/R	CGGAACGACTCGCCTCTT
qseBC-interior-F/R	AAATCGCCAAAATCAATAGCGGTAG
apfA-exterior-F/R	AATACCGGCTCAATTCGCTTA
apfA-exterior-F/R	ACCAGTCACGTAAAGGGTCGGC
apfA-interior-F/R	GGTGTCATCGTATAACCGTATT
apfA-interior-F/R	TTGGCGGCATCGGCTTCTT
apfB-exterior-F/R	GTTGGATACGGATCTGTGAATAG
apfB-exterior-F/R	TTACGGTGGAAGGCGGAACGATTAC
apfB-interior-F/R	CGCATCATTGGTATGTAAGGTC
apfB-interior-F/R	GGATTTCCGTGTTTCGAGTTT
apfC-exterior-F/R	AGGAATACTTCGCTTACACCAC
apfC-exterior-F/R	GTTATCACGGCTACAGAGGC
apfC-interior-F/R	CTAATCGCCCGCTTTGTT
apfC-interior-F/R	CTGAATGCCGCAGTCCCT
apfD-exterior-F/R	CATCATCGGCATAACGCAGTC
apfD-exterior-F/R	AGGCGGATTAGTCGGAGGC
apfD-interior-F/R	GCATAAACCCAGCATACCA
apfD-interior-F/R	TTCCTCTGCTTGCCTTTG
For complement construction
qseBC-F/R	AACTGCAGTTGCAAAATTTCAGCAAATTTAGAC
qseBC-F/R	TTGCGGCCGCTTAGAAGATAATTTCCGCTTTTAAA
apfA-F/R	CCCTCGAGATGCAAAAACTAAGTCTTATTCGAC
apfA-F/R	TTGCGGCCGCTTAATTTGATGCGCAGAAATTTGCC
apfB-F/R	CCCTCGAGATGCAGTATTCCGTATGTGATGTAA
apfB-F/R	TTGCGGCCGCTCATACACTTTCAACATGACCTAAC
apfC-F/R	CCCTCGAGATGTTGAAAGTGTATGAATTTCATT
apfC-F/R	TTGCGGCCGCCTAACCGCCTCCGACTAATCCGCCT
apfD-F/R	CCCTCGAGATGAGTTCTCCGATTTTCGCATGGT
apfD-F/R	TTGCGGCCGCTTAGATATAAATTTTATCAATAATT
For RT-qPCR
16S rRNA-F/R	CCATGCCGCGTGAATGA
16S rRNA-F/R	TTCCTCGCTACCGAAAGAACTT
apfA-qPCR-F/R	GACCGCCGCTACAGTTTTTA
apfA-qPCR-F/R	TTGCCATTTTAGCTACGGTTG
apfB-qPCR-F/R	CGAAGGCGCATCATTGGTAT
apfB-qPCR-F/R	CAAACGCAAATAAACCCGGC
apfC-qPCR-F/R	AAATTTCGCCTAATCGCCCG
apfC-qPCR-F/R	GCCGCAGTCCCTTTAAAACA
apfD-qPCR-F/R	TGTCCCCGCTTCCCAATATT
apfD-qPCR-F/R	TGCTTGCCTTTGGAATTGCT
For protein expression
PqseB-F/R	CGCCCATATGTTGCAAAATTTCAGCAAATTTAGAC
PqseB-F/R	CGCCCTCGAGAGCATCTGAATTTCCTATTTTA
For EMSAs
apf-EMSA-F/R	ATCCCTCTCCTTATCATATCTAAGA
apf-EMSA-F/R	TTTGACCTCTGAATTTATTAAAACG
pilM-EMSA-F/R	CGAAAAATTCTCTTCTGGTAAATGA
pilM-EMSA-F/R	AATTTTAGTTCCTTAAATCAACTTT
glpK-EMSA-F/R	CGTTTCCCTTTTTTACGGACTAAAC
glpK-EMSA-F/R	ATAGTCCTCCTACGGATAACGAAAA

^aRestricton sites are underlined

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Animal Diseases