TABLE 1.

Strains and plasmids used in this study

Strain or plasmidDescriptionaReference or source
Strains
    DH5αF endA1 glnV44 thi-1 recA1 relA1 gyrA96 deoR nupG φ80lacZΔM15 Δ(lacZYA-argF)U169 hsdR17(rK mK+) λLaboratory stock
    MC4100F′ araD139 Δ(argF-lac)U169 rpsL150 (Strr) relA1 flbB5301 deoC1 ptsF25 rbsRLaboratory stock
    CLM24W3110 ΔwaaL16
    BW25113lacIq rrnBT14 ΔlacZWJ16 hsdR514 ΔaraBADAH33 ΔrhaBADLD78, where subscripts indicate strain names12
    BW25113 waaC::KanBW25113 waaC::Kan3
    BW25113 ΔwaaCBW25113 waaC::Kan with kanamycin marker removedThis study
    BW25113 waaL::KanBW25113, waaL::Kan3
    BW25113 ΔwaaLBW25113 waaL::Kan with kanamycin marker removedThis study
    JC80311292 ΔtolRA5
    CE8032 waaL::KanJC8031 waaL::KanThis study
Plasmids
    pACYCpglEntire pgl locus from C. jejuni in pACYC18457
    pACYCpglmutpACYCpgl with W458A and Y459A mutations in the highly conserved region of pglB57
    pACYCpglB::KanpACYCpgl with a Kan insertion in pglB40
    pBAD18Cloning vector, arabinose inducible, Ampr22
    pBAD24Cloning vector, arabinose inducible, Ampr22
    pCP20Ampr and Cmr plasmid with temp-sensitive replication and thermal induction of the yeast Flp recombinase9
    pTrc99ACloning vector, IPTG inducible, AmprAmersham Pharmacia
    pTrc-GT-6×-HisGT acceptor site sequence with C-terminal 6×-His tag in pTrc99AThis study
    pTrc-MBP-GTmalE gene from E. coli in plasmid pTrc-GT-6×-HisThis study
    pET-AcrA-4×-HisacrA gene from C. jejuni with 2 additional glycan acceptor sites in pET22bThis study
    pTrc-ΔspMBP-GTmalE lacking the native signal peptide in plasmid pTrc-GT-6×-HisThis study
    pTrc-spDsbA-MBP-GTmalE with the native signal peptide replaced by the DsbA signal peptide in plasmid pTrc-GT-6×-HisThis study
    pTrc-spTorA-MBP-GTmalE with the native signal peptide replaced by the TorA signal peptide in plasmid pTrc-GT-6×-HisThis study
    pTrc-spMBP-GT-MBPmalE with GT after the native signal peptide in plasmid pTrc-6×-HisThis study
    pTrc-spMBP-GT-MBP-GTmalE with GT after the native signal peptide in plasmid pTrc-GT-6×-HisThis study
    pTrc-spDsbA-TOP7-GTtop7 gene with the DsbA signal peptide in plasmid pTrc-GT-6×-HisThis study
    pTrc-spTorA-GFP-GTgfpmut2 gene with the TorA signal peptide in plasmid pTrc-GT-6×-HisThis study
    pTrc-spDsbA-26.10LC-spPelB-26.10HC-GT26.10 light chain with the DsbA signal peptide followed by the 26.10 heavy chain with the PelB signal peptide in plasmid pTrc-GT-6×-HisThis study
    pTrc-spDsbA-FcIgG1 Fc domain with the DsbA signal peptide in pTrc99AThis study
    pTrc-spDsbA-FcDQNATIgG1 Fc domain with Q295D, Y296Q, and S298A substitutions and the DsbA signal peptide in pTrc99AThis study
    pTrc-spDsbA-AcrA-4×acrA gene from C. jejuni with 2 additional glycan acceptor sites and the native export signal replaced with the signal peptide from E. coli DsbA in pTrc99AThis study
    pTrc-spTorA-AcrA-4×acrA gene from C. jejuni with 2 additional glycan acceptor sites and the native export signal replaced with the signal peptide from E. coli TorA in pTrc99AThis study
    pBAD18-CjaAcjaA gene from C. jejuni with a C-terminal Flag epitope in pBAD18This study
    pBAD24-OmpX*-HisompX gene from E. coli with a C-terminal 6×-His tag in pBAD24; contains unique KpnI and SpeI restriction sites between S53 and S54 of extracellular loop 2 of OmpX44
    pBAD24-OmpX-GTGT acceptor site sequence in pBAD24-OmpX*-HisThis study
    pBAD24-OmpX-GT::PglBBicistronic expression of OmpX-GT and C. jejuni PglB in pBAD24This study
    pBAD18-ClyA-GTclyA gene from E. coli with a C-terminal GT and 6×-His tag in pBAD18This study
    pTrc-YebF-GTyebF gene from E. coli with a C-terminal GT and 6×-His tag in pTrc99AThis study
    pTrc-ΔspYebF-GTyebF gene lacking the N-terminal signal peptide with a C-terminal GT and 6×-His tag in pTrc99AThis study
    pTrc-spYebF-GTN-terminal signal peptide from yebF with a C-terminal GT and 6×-His tag in pTrc99AThis study
    pTrc-YebF-ΔspMBP-GTmalE gene from E. coli lacking the N-terminal signal peptide in pTrc-YebF-GTThis study
    pTrc-ΔspYebF-ΔspMBP-GTmalE gene from E. coli lacking the N-terminal signal peptide in pTrc- ΔspYebF-GTThis study
    pTrc-spYebF-ΔspMBP-GTmalE gene from E. coli lacking the N-terminal signal peptide in pTrc-spYebF-GTThis study
  • a Strr, streptomycin resistant.