Supplementary Table 2. Oligonucleotide Primers Used in This Study

Supplementary Table 2. Oligonucleotide Primers Used in This Study

Supplementary Table 2.Oligonucleotide primers used in this study

Primer name / Sequence (5’-3’)a / Use / Reference
M13RP / GAAACAGCTATGACCATG / R primer binds 3’ cat, used in SOE with F primers / (2)
DAP142 / GGTTCATCGCGTTTTTCC / Ppil::cat PCR screening F primer (used with DAP427 and DAP1263) binds 148bp upstream of the BglII site in theiga gene / C.S. Ryan, unpublished
DAP225 / CGTACAAATGACGGCAGCAACG / Ppil::cat PCR screening F primer (used with DAP427) binds within the spectinomycin cassette / C.S. Ryan, unpublished
DAP427 / TCCAGCTGAACGGTCTGG / Ppil::cat PCR screening R primer (used with DAP142 and DAP225) binds within the cat gene / C.S. Ryan, unpublished
DAP1263 / CGTTTGGATGCCAATGGC / Ppil::cat PCR screening R primer (used with DAP142) binds within the iga gene (downstream of the cat gene) / C.S. Ryan, unpublished
DAP1529 / ATTGGGGAGGGCTATCGTCG / Fluorescent labeled R primer for mapping the transcriptional start point (tsp) for pilH-X / This study
DAP1531 / GTAAACGCGGATACTTCG / Fluorescent labeled R primer for mapping the transcriptional start point (tsp) for pilG/D / This study
DAP1550 / ACCGTCTGAAAACAGCATCG / Fluorescent labeled R primer for mapping the transcriptional start point (tsp) for pilF / This study
DAP1568 / TTACCAATTAACGTCCTATTCTAAGGAAGCTAAAATGGAG / F primer (used with M13RP) to amplify cat from pJKD699 / This study
DAP1569 / CTCCATTTTAGCTTCCTTAGAATAGGACGTTAATTGGTAA / R primer (used with DAP2053) to amplify FA1090 PpilH region and contains sequence of the ribosomal binding site (RBS) and the start codon of cat / This study
DAP1571 / ATTACTTTACGGGGCTAGAACTAAGGAAGCTAAAATGGAG / F primer (used with M13RP) to amplify cat from pJKD699 / This study
DAP1572 / CTCCATTTTAGCTTCCTTAGTTCTAGCCCCGTAAAGTAAT / R primer (used with DAP1573) to amplify FA1090 PpilG region / This study
DAP1573 / TGTAGTCTAACAAACGGC / F primer to amplify FA1090 pilG promoter (PpilG) region and used with PpilGassociated R primers / This study
DAP1574 / TAACAATGAATAAATTTGGGCTAAGGAAGCTAAAATGGAG / F primer (used with M13RP) to amplify cat from pJKD699 / This study
DAP1575 / CTCCATTTTAGCTTCCTTAGCCCAAATTTATTCATTGTTA / R primer (used with DAP1576) to amplify FA1090 and FAM18 PpilF region / This study
DAP1576 / ACGGATAGAACGTATGCC / F primer to amplify FA1090 and FAM18 pilF promoter (PpilF) region and used with PpilF associated R primers / This study
DAP1751 / TATAGTGGATTAACAAAAATCAGG / F primer (used with M13RP) to exclude sequence immediately upstream of CREE in FA1090 PpilH using pJKD3271 as PCR template / This study
DAP1752 / AGGCAGTACGGATAGTACG / F primer (used with M13RP) to amplify FA1090 PpilG without the 5’ CR and the sequence upstream using pJKD3274 as PCR template / This study
DAP1753 / TGACCCGCAACAATATCC / F primer (used with M13RP) to amplify FA1090 PpilG without CREE and the sequence upstream using pJKD3274 as PCR template / This study
DAP1762 / TTCAGCCCGCATATAATTGAAACTAAGGAAGCTAAATGGAG / F primer (used with M13RP) to amplify cat from pJKD3271 / This study
DAP1763 / CTCCATTTAGCTTCCTTAGTTTTCAATTATATGCGGGCTGAA / R primer (used with DAP2053) to amplify the FA1090 PpilH sequence immediately upstream of CREE using pJKD3271 as PCR template / This study
DAP1935 / CGCACCCCCCTGAAAATATAGTGG / F primer (used with M13RP) to introduce P1 mutation (from TATAAT to CCCCCC) and amplify cat using pJKD3271 as PCR template / This study
DAP1936 / CCACTATATTTTCAGGGGGGTGCG / R primer (used with DAP2053) to introduce P1 mutation using pJKD3271 as PCR template / This study
DAP1937 / CAGGCCCCCCAATGAATAAATTTGG / F primer (used with M13RP) to introduce P1 mutation (from TATAAC to CCCCCC) and amplify cat using pJKD3181 as PCR template / This study
DAP1938 / CCAAATTTATTCATTGGGGGGCCTG / R primer (used with DAP1576) to introduce P1 mutation using pJKD3181 as PCR template / This study
DAP1947 / TATGAGGGGGGCTGTACAGG / F primer (used with M13RP) to introduce P2 mutation (from TAAAAT to GGGGGG) and amplify cat using pJKD3181 as PCR template / This study
DAP1948 / CCTGTACAGCCCCCCTCATA / R primer (used with DAP1576) to introduce P2 mutation using pJKD3181 as PCR template / This study
DAP1949 / TCGTTGGGGGGCATGCTTAGAGATTGCCC / F primer (used with M13RP) to introduce P2 mutation (from TAAAAT to GGGGGG) and amplify cat using pJKD3271 as PCR template / This study
DAP1950 / GGGCAATCTCTAAGCATGCCCCCCAACGA / R primer (used with DAP2053) to introduce P2 mutation using pJKD3271 as PCR template / This study
DAP2025 / TGGCGCAAATCGAAAAAAGT / F primer used in quantitative qRT-PCR to amplify recA transcripts / (1)
DAP2026 / GCTGCCGTCCATTTTCATG / R primer used in quantitative qRT-PCR to amplify recA transcripts / (1)
DAP2027 / GACGATTGAGGAACAACGTG / F primer used in quantitative qRT-PCR to amplify rpoD transcripts / C.S. Ryan, unpublished
DAP2028 / GCGTTCTTTGCCCATAATGA / R primer used in quantitative qRT-PCR to amplify rpoD transcripts / C.S. Ryan, unpublished
DAP2053 / TTATTAGGCGGAATCGAGGC / Forward (F) primer to amplify FA1090 and MS11-A pilH promoter (PpilH) region and used with PpilH associated reverse primers (R) / This study
DAP2054 / TACAGTTGACGTCCTATTGCTCTAAGGAAGCTAAAATGGAG / F primer (used with M13RP) to amplify cat from pJKD699 / This study
DAP2055 / CTCCATTTTAGCTTCCTTAGAGCAATAGGACGTCAACTGTA / R primer (used with DAP2053) to amplify MS11-A PpilH region / This study
DAP2056 / TAAGGATCTTGCGCATCCGATTCTGTCC / F primer (used with M13RP) to introduce P1 mutation (from TATAAT to ATCTTG) and amplify cat using pJKD3274 as PCR template / This study
DAP2057 / GGACAGAATCGGATGCGCAAGATCCTTA / R primer (used with DAP1573) to introduce P1 mutation using pJKD3274 as PCR template / This study
DAP2062 / CGACAAAAACGGCAATAAGG / F primer used in quantitative reverse-transcriptase PCR (qRT-PCR) to amplify pilH transcripts / This study
DAP2063 / AACCGAAAGCGATATGG
TG / R primer used in qRT-PCR to amplify pilH transcripts / This study
DAP2068 / ATGCCGCAAATTCAGAAAAC / F primer used in qPCR to amplify pilFtranscripts / This study
DAP2069 / TCATCCTCGACAATGACCAA / R primer used in quantitative qRT-PCR to amplify pilF transcripts / This study
  1. Underlined bases are mutations introduced for the construction Ppil::cat mutants

REFERENCES

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2.Messing, J. 1983. New M13 vectors for cloning. Methods Enzymol. 101:20-78.