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Commit 1c5dcd9c authored by Sichang He's avatar Sichang He
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fix attributions

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......@@ -116,7 +116,7 @@ In charge of outreach, secured funding for the project.</p>
yeast group.
Also assisted in Wiki writing and expert interview.</p>
<h3 id="yunyi-ru"><a class="header" href="#yunyi-ru">Yunyi Ru</a></h3>
<p>In charge of background research, E. <em>coli</em> group lab design, wet labs &amp; wiki
<p>In charge of background research, <em>E. coli</em> group lab design, wet labs &amp; wiki
writing.
Hosted journal clubs and science lectures for human practice.</p>
<h3 id="zhujun-yao"><a class="header" href="#zhujun-yao">Zhujun Yao</a></h3>
......@@ -187,7 +187,7 @@ protein models for candidate microbes via Alphafold and Haddock and evaluated
fidelity of each model via quantiative metrics.
Modelled growth of chosen microbe model computationally.</p>
<h3 id="yuxiang-he"><a class="header" href="#yuxiang-he">Yuxiang He</a></h3>
<p>Built a 3D model for the drug delivery system.</p>
<p>Responsible for building a 3D model of the microcapsule. Created standard pages and provided technical support for the wiki. Provided technical support in using the Gitlab repository. Helped with protein modeling using PyMOL.</p>
<h2 id="advising-support"><a class="header" href="#advising-support">Advising support</a></h2>
<h3 id="linfeng-huang"><a class="header" href="#linfeng-huang">Linfeng Huang</a></h3>
<p>Overall project advising, principal investigator.</p>
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......@@ -103,7 +103,7 @@
<h1 class>Contribution</h1>
<img src="https://static.igem.wiki/teams/4161/wiki/contri-bg.jpg" />
</div>
<p>We designed cell surface display platforms for probiotics including yeast,
<p>We designed cell surface display platforms to display nanobodies on probiotics including yeast,
<em>E.coli</em> Nissle 1917, and <em>L.lactis</em>. To be specific, the yeast Aga2 surface
display platform for nanobodies was developed. We also designed a novel way
to detect protein surface expression by using flow cytometry. Another
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......@@ -110,7 +110,7 @@
<h4 id="summary-of-lab-design"><a class="header" href="#summary-of-lab-design">Summary of Lab Design</a></h4>
<p>In order to decorate the yeast surface with nanobodies against the pathogen, we designed the vector which contains the nanobody gene behind a membrane-sealed protein. The overall experiment design could be divided into 3 phases. In phase I, the expected vector that could potentially express our designed module would be constructed and stored. In phase II, the expression vector would be transformed into yeast and detected for its expression abundance and location. In phase III, the binding affinity and inhibitory efficiency of the pathogen would be tested. Due to the limitation of materials and time under the background of the local pandemic situation, the experiments only accomplished Phases I &amp; II.</p>
<h4 id="genes-of-nanobody-and-pathogen-were-harvested"><a class="header" href="#genes-of-nanobody-and-pathogen-were-harvested">Genes of nanobody and pathogen were harvested</a></h4>
<p>The DNA sequences of nanobody 20ipaD and antigen ipaD were obtained from Sangon in the form of pUC19 plasmid insertions, which were stored inside E. coli. The storage bacteria were amplified, and plasmids containing sequences of interest were extracted with Plasmid mini-prep kit from TIangen. Those plasmids were digested by XhoI and ApaI, and the sequences of interest were recycled by agarose gel recycle kit from Beyotime (<strong>Fig1</strong>).</p>
<p>The DNA sequences of nanobody 20ipaD and antigen ipaD were obtained from Sangon in the form of pUC19 plasmid insertions, which were stored inside <em>E. coli</em>. The storage bacteria were amplified, and plasmids containing sequences of interest were extracted with Plasmid mini-prep kit from TIangen. Those plasmids were digested by XhoI and ApaI, and the sequences of interest were recycled by agarose gel recycle kit from Beyotime (<strong>Fig1</strong>).</p>
<h4 id="recombinant-plasmids-were-constructed"><a class="header" href="#recombinant-plasmids-were-constructed">Recombinant plasmids were constructed</a></h4>
<p>The purified ipaD and 20ipaD DNA segments mentioned in the previous section, along with the pYD1 vector digested by XhoI &amp; ApaI, were ligated, and transformed into DH5α cells that were selected by ampicillin. Colony direct PCR was applied to inspect the plasmid condition of the colonies formed. pYD1-forward and pyd1-reverse primers were used to show the size of Aga2-Protein, thus verifying the sequence between the primer pairs (<strong>Table 1</strong>). The size of pYD1-ipaD plasmid primer box should be about 1500bp, while the size of pYD1-20ipaD should be about 1000bp. As shown in <strong>Fig2 a</strong>, samples &quot;2-3&quot;, &quot;2-5&quot; meet the requirement for pYD1-ipaD, so the bacteria in those colonies contain the recombinant plasmid pYD1-ipaD. Additionally, all samples in lanes 1~10 met the requirement for pYD1-ipaD, which indicates the bacteria in those colonies contain pYD1-20ipad recombinant plasmids (<strong>Fig2 b</strong>). Those colonies were amplified, and the plasmids were extracted. Sequencing data further supported the successful construction of the recombinant plasmids (<strong>Fig3</strong>).</p>
<p><img src="https://static.igem.wiki/teams/4161/wiki/y-table1.jpeg" alt="Table1" /></p>
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......@@ -157,7 +157,7 @@ spending) (UN, 2022).</p>
mainly focused on a novel method for battling antibiotic-resistant pathogens by
<strong>displaying nanobodies on the surface of the probiotics</strong>. While meeting the
goal, our laboratory team did various ways (using three different cells for
trial: E. coli, Yeast and <em>Lactococcus lactis</em>) to achieve and increase the
trial: <em>E. coli</em>, Yeast and <em>Lactococcus lactis</em>) to achieve and increase the
possibility of success. Additionally, our team collaborates with other iGEM
teams from developing countries to contribute to scientific advancement there.</p>
<h2 id="goal-17-partnerships-for-the-goals"><a class="header" href="#goal-17-partnerships-for-the-goals">Goal 17| Partnerships for the goals</a></h2>
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