TABLE 1

Bacterial plasmids and strains used in this study

Plasmid or strainDescriptionaSource or reference
Plasmids
    pGEM-TCloning vector, AmprPromega
    pUC19Cloning vector, AmprNew England BioLabs
    pRY107E. coli-C. jejuni shuttle vector, Kanr39
    RK212.1Helper plasmid for biparental conjugation40
    pCjRpUC19 derivative containing C. jejuni ribosomal site, AmprThis study
    PCjR-MCSpCjR derivative with MCS, AmprThis study
    pROpCjR-MCS derivative with Cj0299 overexpression promoter, AmprThis study
    pROCpRO derivative with chloramphenicol resistance marker, Ampr CmrThis study
    pROC-CjeIpROC derivative with CjeI ORF (rrs::cjeI-cat), Ampr CmrThis study
    pET-21bC-6×His fusion recombinant protein expression vector, AmprNovagen
    pET-28bN-6×His and C-6×His fusion protein expression vector, KanrNovagen
    pChuB::ermpUC19 derivative, suicidal vectorThis study
    pCjeI::catpUC19 derivative, suicidal vectorThis study
    pET21b-CjeIExpression construct of CjeI fused with C-6×HisThis study
    pET28b-CjeIExpression construct of CjeI fused with double 6×HisThis study
Strains
    C. jejuni
        JL241NCTC 11168, wild type, LFC38
        JL686CG8486, wild type, with HFC24
        JL1126CG8486, Eryr (chuB::erm), HFCThis study
        JL1128NCTC 11168, Eryr transformant 1, HFC, sequencedThis study
        JL1129NCTC 11168, Eryr transformant 11, HFCThis study
        JL1130NCTC 11168, Eryr transformant 12, HFC, sequencedThis study
        JL1131NCTC 11168, Eryr transformant 13, HFCThis study
        JL1132NCTC 11168, Eryr transformant 14, HFC, sequencedThis study
        JL1133NCTC 11168, Eryr transformant 17, HFC, sequencedThis study
        JL1134NCTC 11168, Eryr transformant 18, HFCThis study
        JL1135NCTC 11168, Eryr transformant 19, HFC, sequencedThis study
        JL1136NCTC 11168, Eryr transformant 20, LFC, sequencedThis study
        JL1137NCTC 11168, Eryr transformant 21, HFC, sequencedThis study
        JL1138NCTC 11168, Eryr transformant 22, LFC, sequencedThis study
        JL1341CG8486, rrs::cjeI-catThis study
        JL1345NCTC 11168, Eryr transformant 11, rrs::cjeI-catThis study
        JL1347NCTC 11168, Eryr transformant 12, rrs::cjeI-catThis study
        JL1160NCTC 11168, cjeI::catThis study
    E. coli
        BL21(DE3)fhuA2 [lon] ompT gal (λDE3) [dcm] ΔhsdS; λDE3 = λ sBamHIo ΔEcoRI-B int::(lacI::PlacUV5::T7 gene 1) i21 Δnin5New England BioLabs
        DH5αF Φ80lacZΔM15 Δ(lacZYA-argF)U169 recA1 endA1 hsdR17(rK mK+) phoA supE44 thi-1 gyrA96 relA1 λInvitrogen
        TOP10F mcrA Δ(mrr-hsdRMS-mcrBC) Φ80lacZΔM15 ΔlacX74 recA1 araD139 Δ(ara leu)7697 galU galK rpsL (Strr) endA1 nupGInvitrogen
        JL837TOP10/pChuB::ermThis study
        JL1116DH5α/RK212.1/pRY107; donor strain for biparental conjugation23
        JL1159TOP10/pCjeI::catThis study
        JL1165BL21(DE3)/pET21b-CjeI, AmprThis study
        JL1166BL21(DE3)/pET28b-CjeI, Kanr, recombinant CjeI expressionThis study
  • a Ampr, ampicillin resistant; MCS, multiple-cloning site; Cmr, chloramphenicol resistant; Strr, streptomycin resistant.