1. 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

2. N terminus modification of AIDA1

3. 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

4. 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

5. CSR by gene design