Fig. 5: SpyTag/SpyCatcher system can be used to generate a versatile tool for functionalized surface-displayed bacteria or Bap amyloid-like fibers.
From: Bacterial biofilm functionalization through Bap amyloid engineering
a Schematic representation of Bap-Spy/rCatcher-GFP labelling mechanism. When the protein partner SpyCatcher fused to GFP is added, a covalent bond is formed between Bap-Spy and rCatcher-GFP resulting in fluorescent labeling of Bap. b Fluorescence images showing localization of Bap in S. aureus Bap-Spy grown in LB and LB-glu media at 37 °C, 200 rpm until exponential phase (EX) (OD = 0.5) and stationary phase (ST) (OD = 5). Cells were probed with purified recombinant rCatcher-GFP and purified rGFP alone (used as a control) followed by nucleic-acid stain Hoechst. The fluorescence of GFP and Hoechst, the combination of both signals (merge panels) and the differential interference contrast (DIC) images are shown. Scale bar of panels represents 5 μm. c Graphs correspond to the mean of the intensity profiles of cross-sections cells (n~30). Gray shadow corresponds to standard deviation of the mean.
