TABLE 1.

Bacterial strains and plasmids used in this study

Strain or plasmidDescriptionSource or reference
Y. enterocolitica
    WA-314Clinical isolate of serotype O8, carrying virulence plasmid pYVO816
    WA-cPlasmid-cured derivative of WA-31416
E. coli
    DH5αendA1 supE44 hsdR17 (rK mK+) thi-1 recA1 gyrA relA1 Δ(lacZYA-argF)U169 (φ80 lacZΔM15)14
    Sm10 λpirthi thr leu tonA lacY supE recA::RP4-2-TC::Mu-Kan (λpir)23
Plasmids
    pUC-A-1pUC13 with 5-kb EcoRI-HindIII insert fragment of pYVO8 carrying yadA (A-1 fragment)23
    pUC-A-NpUC-A-1, bp 568-645 deleted in yadA (corresponding to neck region, aa 190-215)This study
    pUC-A-S1pUC-A-1, bp 685-732 deleted in yadA (corresponding to 1st 15-mer repeat, aa 229-244)This study
    pUC-A-S2pUC-A-1, bp 685-777 deleted in yadA (corresponding to 1st and 2nd 15-mer repeats, aa 229-259)This study
    pUC-A-S3pUC-A-1, bp 685-867 deleted in yadA (corresponding to first four 15-mer repeats, aa 229-289)This study
    pUC-A-S4pUC-A-1, bp 685-990 deleted in yadA (corresponding to all 15-mer repeats, aa 229-330)This study
    pUC-A-HpUC-A-1, bp 88-564 deleted in yadA (corresponding to head region, aa 30-188)This study
    pUC-A-HNS1pUC-A-1, bp 88-732 deleted in yadA (corresponding to head region, neck region, and 1st 15-mer repeat, aa 30-244)This study
    pUC-A-HNS2pUC-A-1, bp 88-777 deleted in yadA (corresponding to head region, neck region, and 1st and 2nd 15-mer repeats, aa 30-259)This study
    pUC-A-HNS3pUC-A-1, bp 88-867 deleted in yadA (corresponding to head region, neck region, and first four 15-mer repeats, aa 30-289)This study
    pUC-A-HNS4pUC-A-1, bp 88-990 deleted in yadA (corresponding to head region, neck region, and all 15-mer repeats, aa 30-330)This study
    pUC-A-HNS5pUC-A-1, bp 88-1056 deleted in yadA (corresponding to head, neck and stalk regions, aa 30-352)This study
    pUC-A-L1pUC-A-1, bp 991-1056 deleted in yadA (corresponding to three 7-mer repeats, aa 331-352)This study
    pUC-A-L2pUC-A-1, bp 1057-1107 deleted in yadA (corresponding to linking region, aa 353-369)This study
    pHP45ΩOrigin of spectinomycin resistance cassetteSanonnetti
    pGP704Suicide vector, R6K2 replicon, requires π protein in trans by λpir-containing host23
    pGPS-A-1pGP704, 1.8-kb Spcr cassette in EcoRI site, carrying yadA as an EcoRI-SphI fragment from pUC-A-123
    pGPS-A-NpGPS, EcoRI-SphI A-N fragment of pUC-A-NThis study
    pGPS-A-S1pGPS, EcoRI-SphI A-S1 fragment of pUC-A-S1This study
    pGPS-A-S2pGPS, EcoRI-SphI A-S2 fragment of pUC-A-S2This study
    pGPS-A-S3pGPS, EcoRI-SphI A-S3 fragment of pUC-A-S3This study
    pGPS-A-S4pGPS, EcoRI-SphI A-S4 fragment of pUC-A-S4This study
    pGPS-A-HpGPS, EcoRI-SphI A-H fragment of pUC-A-HThis study
    pGPS-A-HNS1pGPS, EcoRI-SphI A-HNS1 fragment of pUC-A-HNS1This study
    pGPS-A-HNS2pGPS, EcoRI-SphI A-HNS2 fragment of pUC-A-HNS2This study
    pGPS-A-HNS3pGPS, EcoRI-SphI A-HNS3 fragment of pUC-A-HNS3This study
    pGPS-A-HNS4pGPS, EcoRI-SphI A-HNS4 fragment of pUC-A-HNS4This study
    pGPS-A-HNS5pGPS, EcoRI-SphI A-HNS5 fragment of pUC-A-HNS5This study
    pGPS-A-L1pGPS, EcoRI-SphI A-L1 fragment of pUC-A-L1This study
    pGPS-A-L2pGPS, EcoRI-SphI A-L2 fragment of pUC-A-L2This study
    pGPS-A-N1pGPS-A-HNS1 carrying neck region (yadA bp 568-684) in front of truncated yadA of pGPS-A-HNS1This study
    pGPS-A-N2pGPS-A-HNS2 carrying neck region (yadA bp 568-684) in front of truncated yadA of pGPS-A-HNS2This study
    pGPS-A-N3pGPS-A-HNS3 carrying neck region (yadA bp 568-684) in front of truncated yadA of pGPS-A-HNS3This study
    pGPS-A-N4pGPS-A-HNS4 carrying neck region (yadA bp 568-684) in front of truncated yadA of pGPS-A-HNS4This study
    pGPS-A-HNS2-FlagpGPS-A-HNS2 carrying 32-bp Flag epitope sequence in front of truncated yadA of pGPS-A-HNS2This study
    pGPS-A-HNS3-FlagpGPS-A-HNS3 carrying 32-bp Flag epitope sequence in front of truncated yadA of pGPS-A-HNS3This study
    pGPS-A-HNS4-FlagpGPS-A-HNS4 carrying 32-bp Flag epitope sequence in front of truncated yadA of pGPS-A-HNS4This study
    pGPS-A-HNS5-FlagpGPS-A-HNS5 carrying 32-bp Flag epitope sequence in front of truncated yadA of pGPS-A-HNS5This study
    pYVO8Virulence plasmid of WA-31416
    pYVO8-A-0pYVO8 yadA,Km-GenBlock inserted in PstI sites of yadA by allelic exchange23
    pYVO8-A-1pYVO8-A-0 with integrated pGPS-A-1, wild-type yadA23
    pYVO8-A-Δ29-81pYVO8-A-0 with integrated pGPS-A-A-Δ29-8123
    pYVO8-A-NpYVO8-A-0 with integrated pGPS-A-NThis study
    pYVO8-A-S1pYVO8-A-0 with integrated pGPS-A-S1This study
    pYVO8-A-S2pYVO8-A-0 with integrated pGPS-A-S2This study
    pYVO8-A-S3pYVO8-A-0 with integrated pGPS-A-S3This study
    pYVO8-A-S4pYVO8-A-0 with integrated pGPS-A-S4This study
    pYVO8-A-S5pYVO8-A-0 with integrated pGPS-A-S5This study
    pYVO8-A-HpYVO8-A-0 with integrated pGPS-A-HThis study
    pYVO8-A-HNS1pYVO8-A-0 with integrated pGPS-A-HNS1This study
    pYVO8-A-HNS2pYVO8-A-0 with integrated pGPS-A-HNS2This study
    pYVO8-A-HNS3pYVO8-A-0 with integrated pGPS-A-HNS3This study
    pYVO8-A-HNS4pYVO8-A-0 with integrated pGPS-A-HNS4This study
    pYVO8-A-L1pYVO8-A-0 with integrated pGPS-A-L1This study
    pYVO8-A-L2pYVO8-A-0 with integrated pGPS-A-L2This study
    pYVO8-A-N1pYVO8-A-0 with integrated pGPS-A-N1This study
    pYVO8-A-N2pYVO8-A-0 with integrated pGPS-A-N2This study
    pYVO8-A-N3pYVO8-A-0 with integrated pGPS-A-N3This study
    pYVO8-A-N4pYVO8-A-0 with integrated pGPS-A-N4This study
    pYVO8-A-HNS2-FlagpYVO8-A-0 with integrated pGPS-A-HNS2-FlagThis study
    pYVO8-A-HNS3-FlagpYVO8-A-0 with integrated pGPS-A-HNS3-FlagThis study
    pYVO8-A-HNS4-FlagpYVO8-A-0 with integrated pGPS-A-HNS4-FlagThis study
    pYVO8-A-HNS5-FlagpYVO8-A-0 with integrated pGPS-A-HNS5-FlagThis study