Cell Surface Reengineering (CSR) CSR by PCR product digestion & ligation: Fusion of ds synthetic oligos to the C terminus of OmpA Fusion of ds synthetic oligos to the N terminus of AIDA1 Mistake here with AIDA1 – we did not make additions to the C terminus

N terminus modification of AIDA1

C terminus modification of OmpA Pre-assay plates C terminus addition of his to the ompA gene The top shows the plates prior to separation by cobalt beads Bottom is the beads – whish have the bact containing the ompAhis + sender (with RFP) The ratios (1:100) are the dilutions I made prior to the assay – so there is in fact enrichment. Post-Assay Plates

CSR by gene Design Fusion of tags & randomers to extracellular portion of OmpA loop 1 (loop insertion) PCR product insertion (950 bps) Insertion of ds oligos

CSR by gene design