Figure 2
EstA β-domain mediated surface display of heterologous passenger domains. (A-B) Induced cells of P. stutzeri A15 pEstAβ-mCherry/eGFP/yEVenus/aiiA/aiiB/attM/bla or empty vector (EV) were treated with proteinase K (PK +) or mock-treated (PK -). Samples were analyzed with Western blot using anti-EstA serum (α-EstA), anti-E-tag antibodies (α-E-tag) or anti-GroEL antibodies (α-GroEL). (C-D) Heat modifiability analysis of proteins in the membrane fractions of samples in (A-B). SDS-PAGE samples were incubated at room temperature (RT) or 100°C and analyzed with Western blot using anti-EstA serum (α-EstA). The band with the higher apparent molecular weight, containing the signal peptide, is indicated with an ‘*’. The band with the lower apparent molecular weight, without the signal peptide is indicated with an ‘o’. Molecular weight markers are indicated at the side of the panels.