Figure 2
From: Engineering the Campylobacter jejuni N-glycan to create an effective chicken vaccine
The E. coli cell surface display-based C. jejuni N-glycan vaccine.
(A) Cartoon depicting the O-antigen ligase (WaaL)-dependent addition of the N-glycan structure to the LPS core of E. coli. (B) Western blot of proteinase K digested whole cell lysates of E. coli wzy::kan (pACYC184 (pglmut)), lane 1 and E. coli wzy::kan (pACYC184), lane 2 probed with R1-4 are shown. The formation of the LPS-core-N-glycan molecule is indicated by an arrow. Molecular weight markers (in kDa) are indicated on the left. (C) FACS analysis of 2 × 10*4 cells of E. coli wzy::kan (pACYC184 (pglmut)) in light grey and E. coli wzy::kan (pACYC184) in dark grey. Fluorescent intensity is shown on the x-axis, cell counts (arbitrary units) are shown on the y-axis. (D) The sequence of the LPS core N-glycan fusion product is shown. N-glycan-derived monosaccharide residues are shown in red. For the LPS-core part, only carbohydrate residues that could be assigned by NMR are shown. Capital letters refer to residues as outlined in Supplementary Table 2. (E) NMR spectrum of the purified LPS core-C. jejuni-N-glycan compound. Correlations from anomeric protons (as indicated) are shown. Green, COSY; red, TOCSY; black, ROESY.
