TABLE 1.

Bacterial strains, plasmids, and primers used in this study

Strain, plasmid, or primerRelevant characteristics or sequenceaReference or source
E. coli strains
    EC One ShotHigh-efficiency transformationInvitrogen
    DH5αHigh-efficiency transformation; NalrInvitrogen
    C600E. coli K-1220
    C600::933WC600 lysogenized with Stx2-converting phage 933W20
    C600::ΔtoxC600 lysogenized with phage Δtox; Cmr; GFPThis study
    C600-SSpontaneous Strr C600This study
    C600KC600-S(pBBR1MCS-2); Strr KanrThis study
    C600A::933WC600-S(pBBR1MCS-4) lysogenized with 933W; Strr AmprThis study
    C600G::ΔtoxC600-S(pBBR1MCS-5) lysogenized with Δtox; Cmr Strr GenrThis study
    PT-32E. coli O157:H7; stx1, stx2; isolated from a patientThis study
    FI-4Fecal E. coli isolated from a healthy personThis study
    FI-15Fecal E. coli isolated from a healthy personThis study
    FI-31Fecal E. coli isolated from a healthy personThis study
    FI-29Fecal E. coli isolated from a healthy personThis study
    FI-37Fecal E. coli isolated from a healthy personThis study
Plasmids
    pBluescript KS(+)Cloning vector; AmprStratagene
    pCR2.1TA cloning vector; Ampr KanrInvitrogen
    pGFPuvGFPuv; AmprBD Biosciences Clontech
    pST76-CCloning vector; Cmr24
    pPIR-KTemperature sensitive suicide vector; Kanr24
    pSG039Upstream and downstream regions of stx2A, GFP, Cmr; in vector pPIR-KThis study
    pBBR1MCS-2Cloning vector; Kanr12
    pBBR1MCS-4Cloning vector; Ampr12
    pBBR1MCS-5Cloning vector; Genr12
Primers
    STX-UP (forward)5′-GCGGCCGCACGTGTTGGGGGAGACGTTC (NotI)This study
    STX-UP (reverse)5′-GAATTCAAGCGAGCCTCCTAAATAAATATGG (EcoRI)This study
    STX-DOWN (forward)5′-ATCGATAATTCAGTCAGTTGACACTTGCCTG (ClaI)This study
    STX-DOWN (reverse)5′-GGGCCCGCGGCCGCACGGAACCAGCAGAATGCC (NotI, ApaI)This study
    GFP (forward)5′-GAATTCAAGCGAGCCTGGTAAATAAATATGG (EcoRI)This study
    GFP (reverse)5′-ATCGATTTATTTGTAGAGCTCATCCATGCC (ClaI)This study
    Cmr (forward)5′-ATCGATCAGCATCACCCGACGCACTTTG (ClaI)This study
    Cmr (reverse)5′-ATCGATCAGTGAGGCACCAATAACTGCC (ClaI)This study
    Verification of Δtox (forward)5′-CGAAATCGAATCGAACCATAACCG (upstream of stx2A)This study
    Verification of Δtox (reverse)5′-CCGAAGAAAAACCCAGTAACAGG (in stx2A)This study
  • a Nalr, naladixic acid resistance; Ampr, ampicillin resistance; Genr, gentamicin resistance. Restriction sites introduced for cloning are underlined in the primer sequence and identified in parentheses.