Team:TU Eindhoven/Design/Membrane Anchor Design
From 2014.igem.org
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 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.
![](https://static.igem.org/mediawiki/2014/7/75/TU_Eindhoven_COMPy_design.jpg)
Figure 1. Gene design COMPy (INPNC).
![](https://static.igem.org/mediawiki/2014/c/cb/TU_Eindhoven_COMPx_design.jpg)
Figure 2. Gene design COMPy (CPX)