Skip to content
Snippets Groups Projects
Commit dd1e3f9c authored by changyulve's avatar changyulve
Browse files

update results.html

parent 43786f5f
No related branches found
No related tags found
No related merge requests found
Pipeline #485367 passed
Stage: build
Stage: deploy
Hide whitespace changes
Inline Side-by-side
Showing
with 124 additions and 37 deletions
results.html
+ 124
− 37
View file @ dd1e3f9c
......@@ -31,14 +31,14 @@
<div class="dropdown-menu"><a class="dropdown-item"
href="contribution.html">Contribution</a><a class="dropdown-item"
href="description.html">Description</a>
<a class="dropdown-item" href="design.html">Design</a>
<a class="dropdown-item"
href="engineering.html">Engineering</a><a class="dropdown-item"
<a class="dropdown-item" href="design.html">Design</a>
<a class="dropdown-item" href="engineering.html">Engineering</a><a class="dropdown-item"
href="parts.html">Parts</a><a class="dropdown-item"
href="results.html">Results</a><a class="dropdown-item"
href="experiments.html">Experiments</a><a class="dropdown-item"
href="safety.html">Safety</a></div>
href="safety.html">Safety</a>
</div>
</li>
<li class="nav-item">
<a class="nav-link" href="human-practices.html">Human Practice</a>
......@@ -131,6 +131,30 @@
copper biosensor using endogenous proteins is feasible, revealing the effects of the CueR of
chassis bacteria overlooked by previous work utilizing this promoter.</p>
<figure class="text-center">
<div class="image-container d-flex justify-content-between">
<div class="flex-fill">
<div class="img-wrapper d-flex align-items-center justify-content-center"
style="height: 160px;">
<img class="img-fluid mx-auto product-item-img mb-3 mb-lg-0 rounded"
src="https://static.igem.wiki/teams/5459/wiki/results/image-21.png" />
</div>
<figcaption>successful result of plasmid construction
</figcaption>
</div>
<div class="flex-fill">
<div class="img-wrapper d-flex align-items-center justify-content-center"
style="height: 160px;">
<img class="img-fluid mx-auto product-item-img mb-3 mb-lg-0 rounded"
src="https://static.igem.wiki/teams/5459/wiki/results/image-22.png" />
</div>
<figcaption>photo of plate for gradient dilution Copper of different type of
circuit.</figcaption>
</div>
</div>
</figure>
<figure class="text-center">
<div class="image-container d-flex justify-content-between">
<div class="flex-fill">
......@@ -204,6 +228,25 @@
split between two exogenous proteins (mVenusNB and CueR) at the same time, which affects the
maximal expression of mVenus. </p>
<p>In a finite system like a bacterial cell, the production of exogenous proteins is constrained
by the limited availability of cellular resources, such as energy, ribosomes, and amino
acids. When multiple proteins are expressed simultaneously, such as CueR and mVenusNB in our
system, they compete for these resources. This competition creates a bottleneck in the
intracellular anabolic flow, limiting the amount of each protein that can be produced. As a
result, even though we would expect increasing CueR expression to enhance activation (and
subsequently increase fluorescence), the overall resource limitation actually reduces the
maximal expression of mVenus, which explains the drop in fluorescence.</p>
<p>Rational modeling often assumes that higher expression of activators like CueR will lead to a
linear or predictable increase in downstream outputs. However, this assumption fails in the
context of resource-limited systems, where non-linear effects such as metabolic burden and
competition between exogenous proteins can dominate. This is why rational models may fail to
predict actual system behavior under such conditions.</p>
<p>To address these complexities, high-throughput screening and machine learning (ML) techniques
can be employed. High-throughput screening allows for the rapid testing of multiple
combinations of conditions to capture the emergent behaviors that rational models may
overlook. ML can then analyze these large datasets to identify patterns and relationships
between variables, providing more accurate predictions of how resource competition affects
system performance.</p>
<figure class="text-center">
<div class="image-container d-flex justify-content-between">
<div class="flex-fill d-flex flex-column">
......@@ -222,12 +265,26 @@
to high-coverage copper receptor libraries</span>
<p>We randomly inserted Promoter and RBS libraries into the CueR 5' end to create a plasmid
library with different CueR expression intensities. After cotransforming this plasmid
library with the reporter plasmid, we obtained a Cu(II) biosensor library. We picked 96
strains and measured the kinetic of OD600 and fluorescence at 0uM and 500uM, respectively.
In this library, the fluorescence multiplicity before and after induction at 500uM ranged
from 5- to 10-fold(Fig.5), showing a broad range of the regulation. 8 of 96 strains had a
higher maximum fluorescence after induction than the two systems in sections I & II
(>35,000) under the same parameter conditions(Fig. 6). Our project demonstrates the
library with the reporter plasmid, we obtained a Cu(II) biosensor library. </p>
<figure class="text-center">
<div class="image-container d-flex justify-content-between">
<div class="flex-fill d-flex flex-column">
<div class="img-wrapper d-flex align-items-center justify-content-center">
<img class="img-fluid product-item-img rounded"
src="https://static.igem.wiki/teams/5459/wiki/results/image-23.png"
alt="Time-A.U. curves" />
</div>
<figcaption>plasmid library plates obtained from GGA</figcaption>
</div>
</div>
</figure>
<p>We picked 96 strains and measured the kinetic of OD600 and fluorescence at 0uM and 500uM,
respectively. In this library, the fluorescence multiplicity before and after induction at
500uM ranged from 5- to 10-fold(Fig.5), showing a broad range of the regulation. 8 of 96
strains had a higher maximum fluorescence after induction than the two systems in sections I
& II (>35,000) under the same parameter conditions(Fig. 6). Our project demonstrates the
feasibility of tuning the CueR expression to parameterize the circuit and provides a
biosensor library for Cu(II) ions.</p>
......@@ -259,7 +316,8 @@
</div>
</div>
</div>
<figcaption style="margin-top: -120px;">Figure 5. a, shows the expression levels of 96 different experimental groups in
<figcaption style="margin-top: -120px;">Figure 5. a, shows the expression levels of 96
different experimental groups in
the absence of copper induction, the horizontal coordinate is time and the vertical
coordinate is the relative fluorescence intensity (RFU) obtained by dividing the A.U
value after removing the background by the OD value, which reflects the local expression
......@@ -270,7 +328,7 @@
copper induction.</figcaption>
</figure>
<br /><br />
<figure class="text-center">
......@@ -299,10 +357,18 @@
</div>
</div>
</div>
<figcaption style="margin-top: -120px;">Figure 6. a, shows the expression levels of 96 different experimental groups with 500 uM Cu(II) induction, the horizontal coordinate is time and the vertical coordinate is the relative fluorescence intensity (RFU) obtained by dividing the A.U value after removing the background by the OD value, which reflects the local expression levels of different Promoter RBS combinations, and the individual experimental data and graphs of each set of experiments are shown in the Appendix. b, shows the histogram of RFU peaks for 96 different experimental groups with 500 uM Cu(II) induction. c, shows the heatmap of RFU peaks for 96 different experimental groups with 500 uM Cu(II) induction.</figcaption>
<figcaption style="margin-top: -120px;">Figure 6. a, shows the expression levels of 96
different experimental groups with 500 uM Cu(II) induction, the horizontal coordinate is
time and the vertical coordinate is the relative fluorescence intensity (RFU) obtained
by dividing the A.U value after removing the background by the OD value, which reflects
the local expression levels of different Promoter RBS combinations, and the individual
experimental data and graphs of each set of experiments are shown in the Appendix. b,
shows the histogram of RFU peaks for 96 different experimental groups with 500 uM Cu(II)
induction. c, shows the heatmap of RFU peaks for 96 different experimental groups with
500 uM Cu(II) induction.</figcaption>
</figure>
<br /><br />
<br /><br />
......@@ -324,9 +390,11 @@
</div>
</div>
</div>
<figcaption style="margin-top: -120px;">Figure 7. a, barplot of induced fold-changes of fine-tuned CueR system. b, heatmap of induced fold-changes of fine-tuned CueR system.</figcaption>
<figcaption style="margin-top: -120px;">Figure 7. a, barplot of induced fold-changes of
fine-tuned CueR system. b, heatmap of induced fold-changes of fine-tuned CueR system.
</figcaption>
</figure>
......@@ -358,18 +426,36 @@
of mVenus NB in the bacterial cells for each metal ion treatment. We found that CueR is
highly specific to Cu2+ only.</p>
<figure class="text-center">
<div class="image-container d-flex justify-content-between">
<div class="flex-fill d-flex flex-column">
<div class="img-wrapper d-flex align-items-center justify-content-center">
<img class="img-fluid product-item-img rounded"
src="https://static.igem.wiki/teams/5459/wiki/results/res8.png" />
</div>
<figure class="text-center">
<div class="image-container d-flex justify-content-between">
<div class="flex-fill d-flex flex-column">
<div class="img-wrapper d-flex align-items-center justify-content-center">
<img class="img-fluid product-item-img rounded"
src="https://static.igem.wiki/teams/5459/wiki/results/res8.png" />
</div>
</div>
</div>
<figcaption style="margin-top: -10px;">Figure9. The response of CueR-pCoA system upon
different Cu2+ concentrations over time.</figcaption>
</figure>
<figure class="text-center">
<div class="image-container d-flex justify-content-between">
<div class="flex-fill d-flex flex-column">
<div class="img-wrapper d-flex align-items-center justify-content-center">
<img class="img-fluid product-item-img rounded"
src="https://static.igem.wiki/teams/5459/wiki/results/image-24.png"
alt="Time-A.U. curves" />
</div>
<figcaption>The response of 1 plasmid/ 2 plasmid system to different concentration
of Mg2+ and Zn2+</figcaption>
</div>
<figcaption style="margin-top: -10px;">Figure9. The response of CueR-pCoA system upon different Cu2+ concentrations over time.</figcaption>
</figure>
</div>
</figure>
<span id="s6" class="section-heading-upper h3"><br />Understanding the effect of Cu2+
concentration on E.coli cell growth</span>
......@@ -380,18 +466,19 @@
concentration that &lt;=12.5mM hinders the growth of bacteria. We can take this possibility
into account when working with data collected in the field.</p>
<figure class="text-center">
<div class="image-container d-flex justify-content-between">
<div class="flex-fill d-flex flex-column">
<div class="img-wrapper d-flex align-items-center justify-content-center">
<img class="img-fluid product-item-img rounded"
src="https://static.igem.wiki/teams/5459/wiki/results/res9.png" />
</div>
<figure class="text-center">
<div class="image-container d-flex justify-content-between">
<div class="flex-fill d-flex flex-column">
<div class="img-wrapper d-flex align-items-center justify-content-center">
<img class="img-fluid product-item-img rounded"
src="https://static.igem.wiki/teams/5459/wiki/results/res9.png" />
</div>
</div>
<figcaption style="margin-top: -10px;">Figure9. The response of CueR-pCoA system upon different Cu2+ concentrations over time.</figcaption>
</figure>
</div>
<figcaption style="margin-top: -10px;">Figure9. The response of CueR-pCoA system upon
different Cu2+ concentrations over time.</figcaption>
</figure>
</div>
......
0% or .
You are about to add 0 people to the discussion. Proceed with caution.
Finish editing this message first!
Please register or to comment