Description: Many brainstorming sessions took place from November through February. During those, the team and its academic supervisors discussed and explored many ideas and concept brought forward by the members. This first phase was one of divergence, which enabled us to cover a wide variety of topics and explore possibilities in many biological fields. From late February started a convergence phase during which we deepened our knowledge on the most promising ideas that emerged from the previous phase. Amongst those ideas, we selected a few that were exciting, original and feasible in the spawn of time available. Finally, in March, after short presentations form the team members, we casted a vote to choose the final project. Our project « the Gatekeeper » provides a new secretion method for recombinant proteins produced within the periplasm through the insertion of a phage porin in the outer membrane of E. coli. Following promising results describing the opening regulation mechanism of this viral protein (Spagnuolo et al., 2010) and based on well-established knowledge on secretion in E. coli, our team aims to condition the porin’s opening system to create a gateway between the intra- and extra-cellular environment. Our success would mean getting rid of the cumbersome lysis step in recombinant protein production and could give rise to continuous recombinant protein production. We will use a directed evolution approach to mutate regions known to play a key role in either the gate opening or the opening regulation (identified by Spagnuolo et al., 2010) and subject the resulting mutants to strong specific selection pressure. Through this method we will also attempt to create a regulating system for our gate’s opening/closure.
Collaboration details:
Year: 2016Visit Wiki
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Updated at: 8/9/16