TABLE 1.

Bacterial strains and plasmids used in this study

Strain or plasmidGenotype or characteristicsaReference or source
Strains
    E. coli
        DH5αendA1 hsdR17 (rK mK+) glnV44 thi-1 recA1 gyrA (Nalr) relA Δ(lacIZYA-argF)U169 deoR [φ80dlacΔ(lacZ)M15]54
        CC118λpirΔ(ara-leu) araD ΔlacX74 galE galK phoA20 thi-1 rpsE rpoB argE(Am) recA1 λpir20
        S17-1λpirthi pro hsdR hsdM+ recA λpir43
        CGSC4982galKCGSCb
    V. fischeri
        ES114WT6
        ESR1Rfr14
        KV733Rfr pgm::Tn10This study
        KV828Rfr, carries pPS18 integrated into the chromosomeThis study
        KV1069Rfr pgm::ermThis study
        KV1170Rfr attTn7::seqA-pgm+This study
        KV1171Rfr pgm::Tn10 attTn7::seqA-pgm+This study
        KV1177Rfr ΔpgmThis study
        KV1306Rfr, carries pKV161 integrated into the chromosomeThis study
        KV1358Rfr, ΔgalKThis study
Plasmids
    pBSBlue-white screen cloning vector; AprStratagene
    pCD3pEVS107 (SpeI) + 3-kb NheI fragment from pPS21, contains seqA and pgm inside the Tn7 transposonThis study
    pCNW1pVO8 BamHI + 5-kb galK+ BglII fragmentThis study
    pEVS79pBC + mob; Cmr44
    pEVS104Conjugal helper plasmid (tra trb); Knr44
    pEVS107Tn7 delivery plasmid; Knr EmrEric Stabb
    pKV25pUC19::erm50
    pKV36pUC19::catThis study
    pKV124pBSL181 containing promoterless lacZ and oriR6K within Tn10 ends52
    pKV161pEVS79 HindIII + 2.8-kb galK+ HindIII fragment from pCNW1This study
    pPS811-kb NheI fragment cloned from KV733, contains pgm::Tn10 and flanking DNAThis study
    pPS12pBS (SacI) + 6-kb SacI fragment from pPS8, contains DNA flanking the Tn10 insertionThis study
    pPS15pBS (XbaI) + 2.6-kb NheI fragment from pPS12, contains DNA upstream of pgmThis study
    pPS18pPS15 BglII + 1.1-kb BamHI fragment encoding Cmr from pKV36 (results in fur::cat mutation)This study
    pPS1914-kb NsiI fragment cloned from KV828, contains seqA+ and a portion of pgmThis study
    pPS20pEVS79 (BamHI) + 2.2-kb BamHI/BglII fragment from pPS19, contains a portion of the pgm locusThis study
    pPS2120-kb XbaI fragment cloned from KV828, contains seqA+ and pgm+This study
    pPS24pPS20 SphI/filled + 1.2-kb SmaI/EcoRV fragment encoding Emr from pKV25 (results in a pgm::erm mutation)This study
    pPS27pEVS79 (XbaI) + 3-kb Nhe fragment from pPS21This study
    pPS33pPS20 with two MscI sites internal to pgm deleted (results in a 606-bp deletion)This study
    pRK2013Conjugal helper plasmid (tra trb); Knr10
    pTMB79.6-kb ClaI fragment cloned from KV1306, contains galK+ and flanking DNAThis study
    pTMB9pTMB7 deleted between two AseI sites (results in 250-bp deletion at the 5′ end of galK)This study
    pUX-BF13Encodes Tn7 transposase (tnsABCDE); Apr4
    pVO8Blue-white screen cloning vector; Cmr Emr51
  • a Nalr, nalidixic acid resistance; WT, wild type; Rfr, rifampin resistance; Apr, ampicillin resistance; Knr, kanamycin resistance; Cmr, CHL resistance; Emr, ERY resistance.

  • b CGSC, E. coli Genetic Stock Center, Yale University, New Haven, Conn.