TABLE 1.

Key bacterial plasmids and strains used in this study

Plasmid or strainDescriptionSource or reference
Plasmids
    pGEM-T EasyPCR cloning vector, AmprPromega
    pCApGEM-T Easy containing 1.6-kb N-terminal cfrA fragment, AmprThis study
    pmCApCA with chloramphenicol resistance cassette inserted in cfrA gene, Ampr CmrThis study
    pRY107E. coli-C. jejuni shuttle vector, Kanr51
    pQE-30N-His6 fusion recombinant protein vectorQiagen
    pCFRA-NHISpQE-30 ligated with cfrA segment encoding mature CfrAThis study
    pCfrApRY107 derivative containing a 2.5-kb cfrA open reading frame plus its promoter regionThis study
    pCfrA (K297A)pCfrA derivative with K297A mutation in CfrAThis study
    pCfrA (R327A)pCfrA derivative with R327A mutation in CfrAThis study
    pCfrA (Q271A)pCfrA derivative with Q271A mutation in CfrAThis study
    pCfrA (F337A)pCfrA derivative with F337A mutation in CfrAThis study
C. jejuni strains
        JL10ATCC 33291, human isolateATCC
        JL1215046764, bovine isolate52
        JL36S3B, chicken isolate43
        JL78W42606, human isolate22
        JL81F34078, human isolate22
        JL83M76297, human isolate22
        JL118CVM20088, chicken isolateChickena
        JL241NCTC 11168, human isolate40
        JL24281-176, human isolate6
        JL36481-176 CJJ81176_0471 isogenic mutant, KanrThis study
        JL324JL241 derivative, cfrA::cmThis study
        JL335JL324 containing pCfrAThis study
        JL442JL324 containing pCfrA (K297A)This study
        JL446JL324 containing pCfrA (R327A)This study
        JL494JL324 containing pCfrA (Q271A)This study
        JL495JL324 containing pCfrA (F337A)This study
E. coli strains
        DH5αF φ80dlacZΔM15 Δ(lacZYA-argF)U169 recA1 endA1 hsdR17 (rK mK+) phoA supE44 thi-1 gyrA96 relA1 λInvitrogen
        JM109endA1 recA1 gyrA96 thi hsdR17 (rK mK+) relA1 supE44 Δ(lac-proAB) [F′ traD36 lacIqZΔM15]Promega
        JL48Conjugation helper strain, DH5α containing plasmid RK20131
        JL122E. coli AN102, enterobactin transport mutant for enterobactin purification2
        JL275JM109 containing pCFRA-NHISThis study
        JL334DH5α containing pCfrAThis study
  • a Isolated from chicken breast and kindly provided by Shaohua Zhao (Center for Veterinary Medicine, Food and Drug Administration).