Difference between revisions of "Team:Uppsala/Results"

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   <li>Successfully integrated five genes from FPP to zeaxanthin into the chromosome of E. coli.</li>
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   <li>Successfully integrated five genes from FPP to zeaxanthin into the <a href="https://2017.igem.org/Team:Uppsala/Zea-Strain">chromosome</a> of <i>E. coli</i>.</li>
 
   <li>Successfully transformed the  crocin pathway into the <a href="https://2017.igem.org/Team:Uppsala/Zea-Strain">zeaxanthin strain</a></li>
 
   <li>Successfully transformed the  crocin pathway into the <a href="https://2017.igem.org/Team:Uppsala/Zea-Strain">zeaxanthin strain</a></li>
 
   <li>Extracted zeaxanthin from the zeaxanthin producing <a href="https://2017.igem.org/Team:Uppsala/Zea-Strain">strain</a></li>
 
   <li>Extracted zeaxanthin from the zeaxanthin producing <a href="https://2017.igem.org/Team:Uppsala/Zea-Strain">strain</a></li>
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       <div class="header"> Chromosomal integration: </div>
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       <div class="header"> Step 3: UGTCs2 </div>
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       <div class="header"> Farnesyl Pyrophosphate (FPP) → Zeaxanthin </div>
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       <div class="header"> Crocetin → Crocin </div>
 
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   <li>Successfully integrated five genes from FPP to zeaxanthin into the chromosome of E. coli.</li>
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   <li><a href="https://2017.igem.org/Team:Uppsala/Parts">Biobrick</a> - Coding for the enzyme UGTCs2 with His-tag and Lac-inducible promoter, characterised with correct sequencing!</li>
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   <li>Successfully transformed the crocin pathway into the zeaxanthin strain</li>
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  <li>Homology <a href="https://2017.igem.org/Team:Uppsala/Model">model</a></li>
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   <li>Extracted zeaxanthin from the zeaxanthin producing strain</li>
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   <li><a href="https://2017.igem.org/Team:Uppsala/Model">Molecular dynamics</a> - the model was stable!</li>
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   <li>Successfully combined with pathway and transformed into <a href="https://2017.igem.org/Team:Uppsala/Zea-Strain">zeaxanthin strain</a>!</li>
 
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Revision as of 15:02, 31 October 2017

<!DOCTYPE html> Results

Summary
We created and combined the zeaxanthin producing strain and with a plasmid containing the extended crocin pathway which gave us an E.coli strain including the entire production pathway from FPP to crocin. In the end, we were able to identify, create and extensively characterize the pathway for crafting crocin and confirm that modern production of crocin in E.coli is red-y.
The three enzyme BioBricks in the zeaxanthin-crocin pathway was assembled to one plasmid using 3A assembly(link 3A assembly protocol) and was inserted into the zeaxanthin producing E.coli strain using electroporation(link Electroporation protocol). The color of the colonies changes at each addition of another enzyme construct (another step in the crocin pathway). This is an indication that something is indeed happening with the bacterial production when we introduce our pathway steps, see figure X.
Chromosomal integration:
Farnesyl Pyrophosphate (FPP) → Zeaxanthin
  • Successfully integrated five genes from FPP to zeaxanthin into the chromosome of E. coli.
  • Successfully transformed the crocin pathway into the zeaxanthin strain
  • Extracted zeaxanthin from the zeaxanthin producing strain
Step 1: CaCCD2
Zeaxanthin → Crocetin dialdehyde
  • Biobrick - Coding for the enzyme CaCCD2 with His-tag and Lac-inducible promoter, characterised with correct sequencing!
  • Homology model
  • Molecular dynamics - the model was stable!
  • Successfully combined with pathway and transformed into zeaxanthin strain!
Step 2: CsADH2946
Crocetin dialdehyde → Crocetin
Step 3: UGTCs2
Crocetin → Crocin
  • Biobrick - Coding for the enzyme UGTCs2 with His-tag and Lac-inducible promoter, characterised with correct sequencing!
  • Homology model
  • Molecular dynamics - the model was stable!
  • Successfully combined with pathway and transformed into zeaxanthin strain!