TABLE 1.

Bacterial strains and plasmids used in this study

Strain or plasmidCharacteristic(s)Reference or source
E. coli strains
    DH5αendA1 hsdR17 supE44 thi-1 recA1 gyrA96 relA1 Δ(lacZYA-argF)U169 deoR [φ80dlac Δ(lacZ)M15]50
    HB101F Δ(gpt-proA)62 leu supE44 ara-14 galK2 lacY1 Δ(mcrC-mrr) rpsL20 xyl-5 mtl-1 recA1350
S. flexneri strains
    2457TS. flexneri wild-type serotype 2a60
    SA100S. flexneri wild-type serotype 2a36
    SA101SA100, Vir, source of aerobactin21
    SA240SA100 iucD::kan21
    SM100SA100 StrrStefan Seliger
    SM160SM100 recA::camLaura Runyen-Janecky
    SM166SM100 sitA::cam48
    SM193wSM100 sitA::cam feoB::tmp iucD::kan48, 62
    SM100 LacSM100 with lacZ integrated into chromosomeThis study
    SM166 LacSM166 with lacZ integrated into chromosomeThis study
Plasmids
    pACYC184Plasmid cloning vector6
    pWKS30Plasmid cloning vector58
    pLAFR1Cosmid cloning vector12
    pEG1Cosmid vector pLAFR1 carrying SA100 sit operon and surrounding sequence48
    pEG3pWKS30 carrying 8-kb HindIII fragment from pEG1 including sitABCDThis study
    pAR1219T7 RNA polymerase gene under control of lacUV5 promoter in pBR32210
    pFZ68pACYC184 carrying 4.6-kb EcoRI fragment of pEG1; includes entire sit operonThis study
    pFZ69pACYC184 carrying 6.4-kb EcoRI fragment of pEG1This study
    pFZ70pACYC184 carrying 13-kb EcoRI fragment of pEG1; includes T7 promoter regionThis study
    pFZ74pACYC184 carrying 5.6-kb EcoRV fragment of pFZ70; includes T7 promoter regionThis study
    pFZ76pACYC184 carrying 2-kb EcoRV/DraI fragment of pFZ74; includes T7 promoter regionThis study
    pGRG36Tn7 insertion vector27
    pGRGlac1E. coli lacZ cloned into pGRG36This study