Team:TU Eindhoven/Overview

Overview Click Coli

Below a quick overview is given of the mechanism behind Click Coli. This system enables the covalent bonding of any DBCO functionalized molecule onto protein anchors that are expressed on the outer membrane of E. Coli. If you wish to use this click system, please take a look at our step-by-step manual.

1. Two plasimids are transformed into E. Coli. One contains a orthogonal tRNA synthetase , which is the engineered Methanocaldococcus jannaschii tRNA(Tyr)(CUA)-tyrosyl-tRNA synthetase pair orthogonal to Escherichia coli. [1] The other one contains the sequence that encodes for a mutated outer membrane protein, hereafter named Clickable Outer Membrane Protein (COMP).
2. Copper catalysed [3+2] azide-alkyne cycloaddition
3. Strain promoted [3+2] azide-alkyne cycloaddition