Team:Aalto-Helsinki/Parts
PARTS
PARTS OVERVIEW
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.
BASIC PARTS
Our basic parts include V. natriegens’ components of the twin-arginine translocation (Tat) pathway, two Tat signal peptides for periplasmic transport, as well as two production proteins as proofs of concept for the utilization of the pathway:
BBa_K3016030 – Vibrio natriegens’ TatAB operon
An operon containing the TatA and TatB components of Vibrio natriegens’ twin-arginine translocation (Tat) pathway.
BBa_K3016031 – Vibrio natriegens’ TatC CDS
A CDS containing the TatC component of Vibrio natriegens’ twin-arginine translocation (Tat) pathway.
BBa_K3016100 – Vibrio natriegens’ TorA Tat signal peptide
A twin-arginine (RR) motif containing signal peptide for periplasmic transport of proteins by the twin-arginine translocation (Tat) pathway. Derived from Vibrio natriegens’ torA gene.
BBa_K3016101 – Vibrio natriegens’ Aminotransferase Tat signal peptide
A twin-arginine (RR) motif containing signal peptide for periplasmic transport of proteins by the twin-arginine translocation (Tat) pathway. Derived from Vibrio natriegens’ aminotransferase gene.
BBa_K3016500 – hGH-6xHis CDS
A CDS containing an intronless human growth hormone gene with a C-terminal 6xHis-tag.
BBa_K3016600 – YGFP CDS
A CDS containing YGFP, a slow bleaching variant of green fluorescent protein (GFP).
COMPOSITE PARTS
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.