Figure 1
From: Structural insights from random mutagenesis of Campylobacter jejunioligosaccharyltransferase PglB
Western blot and ELISA analysis of periplasmic glycoproteins. Analysis of periplasmic proteins of EPA-CP5 producing E. coli strains and control strains harbouring two different expression vectors for PglB Cj . (A) Western blot of normalised periplasmic samples, detection of glycosylated protein with anti-CP5 (left panel) and anti-EPA antisera (right panel). Theoretical molecular weight of unglycosylated EPA: 67.4 kDa. (B) Sandwich ELISA analysis of periplasmic samples for EPA-CP5, average values and standard deviations of ≥ 3 replicates, absorbance measured against air after 15 min development time. lcp: low copy number plasmids pGVXN114 (wt) and pGVXN115 (mut), mcp: medium copy number plasmids pGVXN112Kan (wt) and pGVXN113Kan (mut), mut: inactive PglB W458A-D459A, host strain: E. coli CLM24 (pGVXN150, pGVXN345).