Team:TU Eindhoven/Design Anchors

From 2014.igem.org

Project Description

Membrane Anchors

Gene 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.



Figure 1. Gene design COMPy (INPNC)


Figure 2. Gene design COMPx (CPX)