Fig. 2
Schematic depictions of the expression vectors for the CPR (a) and CYP1A2 (b) autotransporter fusion proteins. The expression cassettes consist of the rhamnose inducible promoter (RhaP), the CtxB signal peptide (SP), passenger (CPR: orange, CYP1A2: red), a connecting region (grey) and the C-terminal parts of the autotransporter AIDA-I or EhaA comprising the β1-domain (β1), α-helix (α) and β-barrel. The connecting region between the CPR and the EhaA is composed of TEV, factor Xa and OmpT protease cleavage sites, a PEYFK epitope and a 15 amino acid long flexible glycine/serine (G4S)3 part (pPQ29). For flow cytometry analysis the myc epitope (Myc) was inserted in between the passengers and autotransporter unit yielding pPQ61 (CPR) and pPQ62 (CYP1A2). The N-termini of the fusion proteins are expanded as white arrows to show the DNA- and amino acid sequences. Plasmids for CYP1A2 autotransporter fusion protein expression (b) contained the pBR322 ori and β-lactamase gene as selection marker (bla) and plasmids for CPR autotransporter fusion protein expression (a) the P15A ori and kanamycin resistance (KanR)