Figure 1.

Depiction of chimeric scaffold proteins and expression cassettes. (A) Chimeric protein scaffolds generated as fusions of the CipA type 1 cohesin coh1C3 (green) with the OlpB type 2 cohesin coh2C2 (blue) and cellulose binding domain CBD (grey). Linkers between cohesin domains, the cell anchor, or the CBD are derived from OlpB (black dotted) or CipA (grey). Optional linkers are represented by dotted lines. (B) Chimeric protein scaffolds generated as fusions of CipA type 1 cohesin coh1C3 (green) with the type 2 cohesin of SdbA (purple) and cellulose binding domain CBD (grey). Double lines represent direct fusion of two domains without a linker sequence. (C) Scaffold expression cassettes showing the N-terminal signal peptide from the lactococcal Usp45 secreted protein (sp_Usp45) and the cell wall anchor motif of the M6 protein (cwa_M6). Expression of the cassettes is under the control of the nisA nisin-inducible promoter (P_nisA) and ribosome-binding site (rbs_nisA) from L. lactis. The transcriptional terminators of the rrnB operon (tlt2) and trpA gene (t_trpA) are located upstream and downstream of the expression cassette, respectively. Optional DNA sequences encoding certain modules are surrounded by dotted lines.