Figure 3

EstA β-domain mediated surface display of heterologous passenger domains. (A-B) Induced cells of P. stutzeri A15 pEstA/pEstA*-aiiA/aiiB/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) or anti-GroEL antibodies (α-GroEL). Similar Western blots for P. stutzeri A15 pEstA/pEstA*-aiiA/aiiB/attM/mCherry/eGFP/yEVenus/bla using anti E-tag antibodies are given in Additional file 6: Figures S5-S8. (C) Heat modifiability analysis of proteins in the membrane fractions of pEstA samples in (A-B). SDS-PAGE samples were incubated at room temperature (RT) or 100°C and analyzed with Western blot using anti E-tag antibodies (α-E-tag). Western blots for pEstA-mCherry/eGFP/yEVenus and pEstA* samples are given in Additional file 7: Figure S9-S11. 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 right side of the panels.