Slide Collection

1
Slide Collection

• SKS

2
Techniques

• Fusion of ds-synthetic oligos to the C terminus
• OmpA AIDA1
• Fusion to the N terminus
• AIDA1
• Fusion to extracellular portion of loop 1 (loop
  insertion)
• OmpA

3
Fusion at C terminus Bead Assay (His tag)
Pre-assay plates
Beads
4
AIDA results
5
Loop Insertion

• PCR product digestion ligation
• Primer design
• Digest-Ligate-Transform motif
• Gene design
• Insertion points created for inserting synthetic
  constructs

6
Loop Insertion PCR products
Pre-loop1 OmpA
OmpA Portion with Modified loop1
Complete Plasmid
EP digested vector
7
PCR products
Lane1 Ladder Lane2 1st portion OmpA Lane 3
strep2-OmpA portion 2 Lane 4 6xHis-OmpA portion
2
8
PCR final plasmid as a template
Red line indicates the 1 kb band
9
Gene Design
10
Gene Design Operations
M
M
11
Gene Design Operations
His/strep tags OR randomers
M
M
M
M
12
PCR Results
Red line indicates the 1 kb line (7/8)