Difference between revisions of "Team:Bio Without Borders/Results"

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<h3>Cloning and expression of Factor C </h3>
 
<h3>Cloning and expression of Factor C </h3>
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<p> 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:
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<p> Our first choice was whether to express the Factor C gene from the Atlantic horseshoe crab (<i>Limulus polyphemus</i>) or from the Japanese Horseshoe crab (<i>Tachypleus tridentatus</i>). The latter had been described more fully in the literature. But we wanted to use the one from <b>our</b> homeland. We codon-optimized it for our preferred expression system, <i>Pichia pastoris</i>, 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.
  
 
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<h3> Project Achievements </h3>
 
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<p>You can also include a list of bullet points (and links) of the successes and failures you have had over your summer. It is a quick reference page for the judges to see what you achieved during your summer.</p>
 
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<ul>
 
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<li>A list of linked bullet points of the successful results during your project</li>
 
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<li>A list of linked bullet points of the unsuccessful results during your project. This is about being scientifically honest. If you worked on an area for a long time with no success, tell us so we know where you put your effort.</li>
 
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<h3>Inspiration</h3>
 
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<p>See how other teams presented their results.</p>
 
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<li><a href="https://2014.igem.org/Team:TU_Darmstadt/Results/Pathway">2014 TU Darmstadt </a></li>
 
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<li><a href="https://2014.igem.org/Team:Imperial/Results">2014 Imperial </a></li>
 
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<li><a href="https://2014.igem.org/Team:Paris_Bettencourt/Results">2014 Paris Bettencourt </a></li>
 
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Revision as of 03:42, 18 October 2018

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.