The twin-arginine translocation (Tat) pathway in Vibrio natriegens was the main focus of our iGEM project and our submitted BioBricks reflect this. We identified the genes encoding proteins of the pathway and multiple signal sequences targeting proteins for translocation. Additionally we included two proteins for testing the pathway and as a proof of concept for industrial applications.
The signal peptides can be combined with a wide range of different proteins to facilitate their transport into the periplasm. We have illustrated this with our composite parts TorA-yGFP and TorA-hGH. The Tat pathway components can be overexpressed to increase translocation rates into the periplasm.
Our basic parts include: tatAB operon, tatC CDS, TorA signal peptide, Aminotransferase signal peptide, YGFP, hGH.
Our composite parts include: TorA-YGFP, TorA-hGH.
BBa_K3016210 – TorA-hGH-6xHis
A Composite part of C-terminally 6xHis tagged human growth hormone (hGH) fused with a N-terminal Vibrio natriegens’ TorA signal peptide for periplasmic transport via the twin-arginine translocation (Tat) pathway.
BBa_K3016200 – TorA-YGFP
A Composite part of YGFP fused with a N-terminal Vibrio natriegens’ TorA signal peptide for periplasmic transport via the twin-arginine translocation (Tat) pathway.