Basic Parts

BBa_K2719000: GST Coding site

Glutathione S-transferase (GST) is a peptide derived from Schistosoma japonicum, in which a target protein may be fused on the N-terminal. This fusion allows the purification of the target protein using glutathione affinity and maximize the presence of the protein in soluble state, rather than on inclusion bodies. This tag may be later removed by adding a protease recognition site between GST and the target protein (Harper & Speicher, 2013).

BBa_K2719001: Tenascin Domain V

Tenascin is a human protein that is involved in tissue regeneration processes. The domain V contains heparin binding sites that are fundamental for our scaffold. Also contains multiple growth factor binding sites and, even when we are only working with leptin is important because it would help in the improvement of tissue regeneration. In addition, it has been reported that it participated stem cells signalling.

BBa_K2719002: GST-Tenascin Domain V Fusion Protein

This part is the fusion between both proteins and that helps us to simplify tenascin purification. GST has tenascin fused on the N-terminal. For more information see BBa_K2719001 and BBa_K2719003.

BBa_K2719003: Optimized GST Coding site

GST coding sequence (see part BBa_K2719000) with codon optimization for E.coli BL21 (DE3) to avoid complications in the production of this recombinant protein.

BBa_K2719004: Tenascin Fibronectin Domains

Coding sequence from tenascin fibronectin domains I-V have the property of interacting with the proliferation of fibroblast, but the domain with the main characteristic previously mentioned is domain V (see part BBa_K2719001).

BBa_K2719006: Collagen V- like protein coding site

Collagen V is one of the main proteins involved in the extracellular matrix, thus it would be important in the structure of our scaffold. The main challenge for producing recombinant collagen V is that for its correct structure it requires post-traductional modifications, specially the hydroxylation of proline. To overcome this challenge, we designed this part to incorporate hydroxyproline, a non-canonical amino acid. For its proper incorporation, this part is optimized for its use on E.coli Rosetta (DE3).

BBa_K2719008: Leptin coding site.

This is one of the most important parts because is the growth factor that we are using for accelerating the process of skin regeneration. Is a protein that is found in adipocytes cells on many living beings and it has been discovered that it has a regenerating property, because it has been found in many wound repair processes. It has been shown that it increases glucose intake speeding up metabolism, thus, accelerating cell proliferation.