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               <div class="h3"><strong>Expression of uTP tagged by Fluorescent proteins</strong></div>
               <p>The ultimate goal with the uTP sequences we identified is to understand and confirm whether they are indeed responsible for protein import into UCYN-A. Conventional methods to check this would require a toolbox for genetic manipulation of B. bigelowii, not yet available and beyond the scope of this project. We therefore opted for using 2 model eukaryotes for further research on uTP’s behavior, namely C. reinhardtii and S. cerevisiae, and designed an experiment to confirm uTP’s function without modifying B. bigelowii.</p>
               <p>We worked off of a <em>Saccharomyces</em> and a <em>Chlamydomonas</em> backbone, pUDE1311 and pOpt2-mVenusBle respectively, in order to design constructs expressing 
-              fluorescent proteins (FP) tagged by known transit peptides as well as uTP. Unmodified, pUDE1311 expresses ymNeongreen and pOpt2-mVenusBle expresses mVenus, a YFP analogue; Both express AmpR for selection on <em>E.coli</em>, while . We designed 2 constructs for expression in our yeast and 3 in our algae. For our yeast, one construct had uTP inserted in the C-terminus of ymNeongreen and the other had MTS1, a mitochondrial transit peptide <a href="#cite2" style="color: #185A4F;">[2]</a>, inserted in the N-terminus of the fluorescent protein.
+              fluorescent proteins (FP) tagged by known transit peptides as well as uTP. Unmodified, pUDE1311 expresses ymNeongreen and pOpt2-mVenusBle expresses mVenus, a YFP analogue; Both express AmpR for selection on <em>E.coli</em>, and while pOpt expresses a Zeocin resistance gene for selection on <em>C. reinhardtii</em> pUDE expresses URA3 for auxotrophic selection on <em>S. cerevisiae</em> CEN.PK 113-5D, an strain with uracil knockout. We designed 2 constructs for expression in our yeast and 3 in our algae. For our yeast, one construct had uTP inserted in the C-terminus of ymNeongreen and the other had MTS1, a mitochondrial transit peptide <a href="#cite2" style="color: #185A4F;">[2]</a>, inserted in the N-terminus of the fluorescent protein.
               For our algae, one construct had uTP inserted in the C-terminus of mVenus, while the two others had a chloroplastic (cTP, <a href="#cite3" style="color: #185A4F;">[3]</a>) and a mitochondrial transit peptide (mTP, <a href="#cite11" style="color: #185A4F;">[11]</a>) respectively, both inserted in the N-terminus of mVenus. Plasmid maps for our constructs and vectors can be found in our Materials and Methods page.
               We planned to observe the localization of uTP in the absence of UCYN-A in these species, hypothesizing based on the dry-lab analysis detailed above that we would observe uniform diffusion in the cytoplasm. Cells transformed with the regular pUDE and pOpt plasmids as well as the known transit peptides would serve as controls showing both uniform diffusion as well as localization to organelles respectively.
               </p>
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             <div class="img-pagestyle">
-              <img src="https://static.igem.wiki/teams/5054/experiment-abstract.jpeg.png" alt="Fig 1: Graphical overview of the experiment plan.">
+              <img src="https://static.igem.wiki/teams/5054/experiment-abstract.jpeg" alt="Fig 1: Graphical overview of the experiment plan.">
             <figcaption>Figure 9: Example of a diagnostic colony PCR gel: Here,
             </figcaption>
             </div>