Fig. 4

Co-overexpression of BAM improves surface display of DTS trimeric autotransporter UpaG chimeras. A, B Schematic representation of UpaG-SpT2-Calm (A) and UpaG-GFPnb (B) and Western blots of whole cell lysates to analyze their expression in BL21 (DE3). The HA tag, SpT2 tag and calmodulin domains in (A) and GFPnb in (B) were inserted at the N-terminus of the truncated passenger domain (UpaG position are given). The UpAG bands were detected by incubating the Westernblots with anti-HA. The putative positions of monomeric (*), dimeric (**) and trimeric (***) complexes [45] are indicated on the right side of the panels. C Fold change of surface displayed UpaG- -SpT2-Calm, as indicated by flow-cytometric analysis of binding of SpC2-mScarlet, when expressed with or without overproduced BAM. D Fold change of surface displayed UpaG-GFPnb, as indicated by flow cytometric analysis of binding of GFP, when expressed with or without overproduced BAM. ***p < 0.005. Data presented is the mean of three independent experiments