Team:Groningen/Project/secretion

From 2013.igem.org

Silk production

We developed 12 different types of spider silks (figure 1). We have one spider silk protein with a N and C terminus, a spider silk without a N and C terminus and a spidersilk without one block (explain the difference). A codon optimization script was developed to codon optimize the spider silk for b. subtilis. Codon optimization (state why we did codon optimization and link towards the codon optimazation page on the wiki for all the details). Every silk gene was codon optimized twice with different results. All the codon optimized silk genes were ordered and made synthetically (figure 2).



Figure 1, Major ampullate Spidroin 2 (MaSp2) from Argiope aurantia

Figure 2, Our constructed silk. All parts are made into biobricks and can be combined together.

The silk genes are provided with an signal peptide and a strep tag. The signal peptide will ensure the secretion of silk by the natural pathways of b. subtilis (figure 3). The strep tag will be used for our coating mechanism.
Figure 3: the secretion pathway of silk