Team:Bio Without Borders/Results

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Results

Here you can describe the results of your project and your future plans.

What should this page contain?

• We succeeded in cloning the two proteins that make up the test system: Factor C and our substrate (GFP-linker-CDB-CBD)
• We submitted them to the iGEM registry in pSB1C3.
• We attempted to express Factor C in Pichia pastoris, a yeast expression system.
• We attempted to express the substrate in E. coli.

Cloning and expression of Factor C

Our first choice was whether to express the Factor C gene from the Atlantic horseshoe crab (Limulus polyphemus) or from the Japanese Horseshoe crab (Tachypleus tridentatus). The latter had been described more fully in the literature. But we wanted to use the one from our homeland. We codon-optimized it for our preferred expression system, Pichia pastoris, and sent it for synthesis. Unfortunately, after several failed attempts, IDT was unable to synthesize it. So we switched to the Japanese horseshoe crab and had it synthesized as G-blocks for later assembly, and also as a complete gene. Our attempts at G-block assembly failed, and we ended up using the full gene synthesized in the plasmid pIDT.