Team:Dundee/Project/ProductionExport
From 2013.igem.org
Production & Export
Introduction
The ToxiMop is an engineered E. coli bacteria that expresses PP1 and can be used as a molecular mop to remove microcystin from contaminated water. Central to successfully engineering this machine was PP1 production and export. This is because microcystin binds to PP1 in the periplasm.
We explored both the Twin Arginine Translocase (Tat) pathway and Secretory (Sec) pathway as potential export mechanisms. However initial Western blot results indicated that PP1 was exported into the periplasm more successfully via the Tat pathway .Therefore production and export, based on Tat transportation, was selected as a modelling focus to allow us to optimise the construction of our prototype ToxiMop.
Building a Model for Tat Transport
The Tat machinery is a biological pathway that transports folded proteins from the cytoplasm into the periplasm. It consists of three small membrane proteins; TatA, TatB and TatC.