Journal: Journal of Virology
Article Title: Polyvalent phage GSP004 recognizes O-antigen polysaccharide receptors in Salmonella and Escherichia coli through tail fiber protein ORF208
doi: 10.1128/jvi.00810-25
Figure Lengend Snippet: Prediction and identification of the RBP from phage GSP004 . ( A ) Purification of recombinant ORF208 protein analyzed by SDS-PAGE with Coomassie blue staining. Lane M, Precision Plus Protein Dual Color Protein Marker (Bio-Rad). Lane 1, unpurified recombinant ORF208 protein. Lane 2, Ni-NTA affinity chromatography purified recombinant ORF208 protein. ( B ) Competitive binding assay of ORF208 protein to host bacteria. Pre-incubation of S. Enteritidis SE006 or E. coli ATCC 35150 with ORF208 reduced phage adsorption. Conversely, the addition of LPS restored adsorption by competing with ORF208 binding. ( C ) Antibody blocking assay. Pre-incubation of phage GSP004 with anti-ORF208 antibody significantly inhibited bacterial infectivity. ( D ) Fluorescence microscopy analysis of EGFP-tagged ORF208 binding to wild-type, Δ rfaL mutants, and Δ rfaL -complemented strains. EGFP-tagged ORF208 binds to wild-type and complemented strains, but not to Δ rfaL mutant. * P < 0.05, *** P < 0.001.
Article Snippet: LPS was extracted from S . Enteritidis SE006 or E. coli ATCC 35150 using the LPS Extraction Kit (Bestbio, Shanghai, China) for subsequent experiments.
Techniques: Purification, Recombinant, SDS Page, Staining, Marker, Affinity Chromatography, Competitive Binding Assay, Bacteria, Incubation, Adsorption, Binding Assay, Antibody Blocking Assay, Infection, Fluorescence, Microscopy, Mutagenesis