Fig. 5: Nanobody CAM targeting of E. coli integral membrane proteins OmpA and OmpC.
From: A whole-cell platform for discovering synthetic cell adhesion molecules in bacteria
a The amino acid sequence of NbompA and NbompC. CDR1 (green), CDR2 (red), and CDR3 (blue) are indicated. b Self-aggregation of E. coli expressing NbompA and NbompC. Cultures underwent 24 h aTc induction before settling. Cultures that did not undergo induction (i.e., not expressing nanobodies) are presented as controls. c, d Conjugation frequency of pGenR under the liquid growth condition. Recipient cells expressing indicated nanobodies were co-cultured with S17-1 donor cells expressing (c) or not expressing (d) the indicated antigens. Data are presented as means ± SD. n = 3 biological replicates with three technical replicates each. Asterisks indicate statistically significant differences between the indicated mean values (p < 0.05, two-tailed t-test). Fluorescence micrographs showing the antigen-binding specificity of NbompA-GFP (e) and NbompC-GFP (f). NbompA-GFP and NbompC-GFP were incubated with wild-type E. coli and the antigen-deleted strains (∆ompA or ∆ompC) for 1 h before observation. Scale bar = 1 μm. Full-size images are available in Supplementary Fig. 9. The experiments were repeated three times independently with similar results. Source data are provided as a Source Data file.
