S. aureus was exposed to gamma radiation at an irradiation dose of 100 Gy, and its descendant were cultured under normal conditions. Protein identification and quantification of unirradiated, irradiated, and descendant of irradiated S. aureus were performed using DIA proteomic technology to obtain protein abundance profiles for each group. Combined with biological experiments, further functional enrichment analysis and protein-protein interaction network analysis were performed to explore the effect of gamma radiation on the pathogenicity of S. aureus.