Team:TU Eindhoven/Design/Membrane Anchor Design

Membrane Anchor Design

In short, both membrane proteins had a TAG codon introduced for the non-natural amino acid and a HA-tag for detection methods. For INPNC the TAG codon and the HA-tag were introduced at the C terminus of the protein (Figure 1). The CPX proteins already contained a peptide library part at the N-terminus so the TAG codon was introduced in that part. This has been done by using Site Directed Mutagenesis. The HA-tag was introduced at the C-terminus (Figure 2). After modification the proteins were renamed to Clickable Outer Membrane Protein x (COMPx), originally CPX, and COMPy, originally INPNC. Protocols for creation of these anchors can be found on the protocol page.

Figure 1. Gene design COMPy (INPNC).

Figure 2. Gene design COMPy (CPX)