1. Techniques Fusion of ds synthetic oligos to the C terminus of OmpA
Fusion of ds synthetic oligos to the N terminus of AIDA1
Fusion of tags & randomers to extracellular portion of OmpA loop 1 (loop insertion)
Mistake here with AIDA1 – we did not make additions to the C terminus

2. Fusion of tags to N-terminus of AIDA-1

3. Fusion at C terminus: Bead Assay (His tag)
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.
Beads

4. Loop Insertion PCR product digestion & ligation Gene design
Primer design
Digest-Ligate-Transform motif
Gene design
Created insertion points for compatible synthetic constructs