Team:Groningen/Project/secretion

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Silk production


Figure 2: schematic drawing of natural silk

The spider silk we used is the so called dragline silk from the Argiope aurantia (MaSp2) from Brookes et al (2008). It is the strongest silk produced inside the spider. The spider silk consists out of a big repetitive domain (around 2500 base pairs) with an N and a C terminus (figure 2 b). (write something about figure 2a as well of crop the image).

Our spider silk

We developed 12 different types of spider silk (Link towards biobricks and check the number) (figure 3). 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 4).




Figure 3, our spider silks


Figure 4: Not sure if this image needs to be on our site, if so please explain thoroughly. 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 5). The strep tag will be used for our coating mechanism.


Figure 5: the secretion pathway of silk


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   Silk
   Silk Secretion
   Coating Mechanism
   Backbone

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