FIGURE 3: The Coa:msfGFP fusion protein was successfully secreted from S. aureus and functioned correctly. (A) n-gel fluorescence of GFP/msfGFP in a native PAGE gel containing supernatants from the S. aureus wildtype parent strain, as well as S. aureus wildtype and S. aureus Δvwbp both expressing Coa:msfGFP. His-tagged GFP and supernatant from the strain expressing Sec:msfGFP were also loaded to the gel to serve as a molecular marker and as a positive control for GFP and msfGFP fluorescence, respectively. The gel shows that Coa:msfGFP was secreted as an intact, fluorescent protein, giving a band at the expected weight of msfGFP and Coa combined [53]. (B) Coagulation of S. aureus 29213 wildtype and Δvwbp producing either Coa:msfGFP or unmodified Coa and S. aureus 29213 ΔcoaΔvwbp after 24 hours incubation with human plasma at 37°C. All strains producing Coa coagulated plasma, while the double mutant ΔcoaΔvwbp did not.

53. Boden MK and Flock JI (1989). Fibrinogen-binding protein/clumping factor from Staphylococcus aureus. Infec Immun 57(8): 2358-2363. 10.1128/iai.57.8.2358-2363.1989