TABLE 1.

Strains, plasmids, and oligonucleotide primers used in this study

Strain, plasmid, or primerRelevant characteristicsReference or source
V. vulnificus strains
    ATCC 29307Type strain12
    ARATCC 29307 but spontaneous rifampin resistanceThis study
    MO6-24/0Clinical isolate48
    CN7Clinical isolateThis study
    CNUH94-4Clinical isolateThis study
    SC9649Environmental isolate (seawater)This study
    SC9720Environmental isolate (sediment)This study
    SC97126Environmental isolate (seafood)This study
    KPR101AR but ΔrpoS (allelic exchange with pKP13)This study
E. coli strains
    DH5αφ80dlacZΔM15 recA1 endA1 gyrA96 relA1 thi-1 hsdR17(rK mK) supE44 deoR Δ(lacZYA-argF)U169Laboratory collection
    JM109endA1 recA1 gyrA96 thi-1 hsdR17(rK mK) relA1 supE44 Δ(lac-proAB) [F′ traD36 proAB lacIqZΔM15]Promega
    SM10λpirthi-1 thr leu tonA lacY supE recA::Rp4-2-Tc::Mu λpir; Kmr34
    AMS6K-12 (λ F Δlac)31
    ZK918W3110 ΔlacU169 tna-2 rpos::kan bolA:lacZ2
Plasmids
    pUC19Cloning vector; lacZα Apr49
    pINE32pUC19 with 2.75-kb Sau3AI fragment of V. vulnificus DNA containing the complete coding sequence of V. vulnificus of rpoSThis study
    pKP11pINE32 with deletion of 762-bp NruI fragment (V. vulnificus ΔrpoS)This study
    pDM4Suicide vector; oriR6K Cmr23
    pKP13pDM4 containing SmaI and XbaI fragment of pKP11This study
    pLAFR5IncP Tcr; derivative of pLAFR3 containing a double cos cassette14
    pKP14pLAFR5 containing EcoRI and HindIII fragment of pINE32This study
    pQE30Expression vectorQiagen
    pQErpoSpET containg 1,072-bp PCR-amplified V. vulnificus rpoS coding regionThis study
    pHK0011pRK415 with promoterless luxAB; Tcr12
    pHL-03katGvv::luxAB transcriptional fusion in pHK0011This study
    pRK415IncP ori; broad-host-range vector; oriT of RP4; Tcr14
    pKP51pRK415 containing KpnI and HindIII fragment of V. vulnificus katGThis study
Primers
    rpoSvv-F5′-CGGATCCAGTATCAGCAACACAGTCACCAAAG-3′
    rpoSvv-R5′-GGCCAAGCTTCTATGATTCCATAGCCTTTTTTC-3′
    F25′-AACGCGCTAAGAGGTTATGGC-3′
    R25′-AGCAAGAACGGTGGATATCGC-3′
    katG-F-KpnI5′-GTTGGTACCTCAACTACCGC-3′
    katG-R-XbaI5′-GGATGTCTAGATTGAGGGCC-3′
    katG-R-HindIII5′-GCAAGTCTGATAAAAGCTTCG-3′