From c0944a039c48e8c35b2d4ebe9315508cde58f1d9 Mon Sep 17 00:00:00 2001
From: lovisongjie <q2516581@126.com>
Date: Tue, 1 Oct 2024 23:21:51 +0800
Subject: [PATCH] design fix and engineering fix
---
design.html | 2 +-
engineering.html | 38 +++++++++++++++++++++++++++++++-------
2 files changed, 32 insertions(+), 8 deletions(-)
diff --git a/design.html b/design.html
index f2216a7..0579b03 100644
--- a/design.html
+++ b/design.html
@@ -68,7 +68,7 @@
<section class="clean-block clean-post dark">
<div class="container">
<div class="block-content">
- <div class="post-image" style="background-image:url('assets/img/scenery/image3.jpg');"></div>
+ <div class="post-image" style="background-image:url('https://static.igem.wiki/teams/5459/design.jpg');"></div>
<div class="post-body">
<h2 class="section-heading mb-4">
<span class="section-heading-lower"><strong>Design</strong></span>
diff --git a/engineering.html b/engineering.html
index 817eb17..19081f3 100644
--- a/engineering.html
+++ b/engineering.html
@@ -84,12 +84,12 @@
important role in regulating copper ion homeostasis and resistance to copper toxicity. When
the external copper ion concentration increases, CueR binds to copper ions with high
affinity. Once copper ions enter the cell, CueR rapidly binds to them, triggering a
- conformational change in the CueR protein.</p>
+ conformational change in the CueR protein[1,2].</p>
<p>In the absence of copper ions, CueR binds to the promoter region (pCoA) in a repressive
state, preventing the expression of downstream genes. When CueR binds to copper ions, it
undergoes a conformational change, transitioning from a repressive state to an active state,
- which then initiates the expression of downstream genes. These genes include:</p>
+ which then initiates the expression of downstream genes. These genes include[3]:</p>
<li>CopA: Encodes a copper ion-transporting ATPase that expels excess copper ions from the cell,
maintaining copper ion balance. </li>
@@ -107,7 +107,7 @@
</ul>
</li>
<li>2. The second plasmid contains a CueR-sensitive pCoA promoter, with a downstream encoded
- fluorescent protein:
+ fluorescent protein[4]:
<ul>
<li>CopA -BBa_B0030- mVenusNB - BBa_B1005</li>
</ul>
@@ -130,9 +130,9 @@
<span id="s11" class="section-heading-upper h4"><br />Design</span>
<p>Copper efflux regulator (CueR) is a Cu+- and Ag+- sensing metalloregulator in E.coli that
controls the expression of two genes involved in metal homeostasis: CopA, which encodes a
- copper/silver ATPase, and CueO, which encodes a copper oxidase [1]. Like most other
+ copper/silver ATPase, and CueO, which encodes a copper oxidase [5]. Like most other
metalloregulators, CueR acts on promoter DNA that exceeds the optimal length (~17bp) for
- recognition by sigma70, a subunit of RNA polymerase (RNAP) [2,3]. Taking advantage of this
+ recognition by sigma70, a subunit of RNA polymerase (RNAP) [5,6]. Taking advantage of this
endogenous Cu(II) sensor system, we designed a simplified prototype plasmid to measure the
concentration of Cu(II) ions. </p>
<p>This reporting plasmid contains a CueR-regulated copper inducible promoter (BBa_K190017)
@@ -150,7 +150,7 @@
<p>We monitored the growth status and fluorescence intensity of the bacterial culture in this
expression system at different copper ion concentrations, as shown in the figure.
Additionally, after the system reached a steady state, we recorded the final fluorescence
- values and plotted a response curve against the different copper ion concentrations.</p>
+ values and plotted a response curve against the different copper ion concentrations[7].</p>
<figure class="text-center">
<div class="image-container d-flex justify-content-between">
@@ -384,7 +384,31 @@
this model can be consulted on our provided "Model" page. Through this model, we aim to
reveal the specific mechanisms by which promoter sequence characteristics affect their
initiation activity, providing a theoretical foundation and experimental guidance for
- subsequent research on gene expression regulation.</p>
+ subsequent research on gene expression regulation.
+ </p>
+
+
+ <span id="References" class="section-heading-upper h3"><br />References</span>
+ <span>
+ <br />
+ [1] Hu, Y. & Liu, B., 2024. The copper efflux regulator (CueR). Subcellular Biochemistry, 104, pp.17-31. doi:10.1007/978-3-031-58843-3_2.
+ <br /><br />
+ [2] Checa, S.K. & Soncini, F.C., 2011. Bacterial gold sensing and resistance. Biometals, 24(3), pp.419-427. doi:10.1007/s10534-010-9393-2.
+ <br /><br />
+ [3] Zhou, X., Xiang, Q., Wu, Y., et al., 2024. A low-cost and eco-friendly recombinant protein expression system using copper-containing industrial wastewater. Frontiers in Microbiology, 15, p.1367583. Published on 21 March 2024. doi:10.3389/fmicb.2024.1367583.
+ <br /><br />
+ [4] Lischik, C.Q., Adelmann, L. & Wittbrodt, J., 2019. Enhanced in vivo-imaging in medaka by optimized anaesthesia, fluorescent protein selection and removal of pigmentation. PLoS One, 14(3), p.e0212956. Published on 7 March 2019. doi:10.1371/journal.pone.0212956.
+ <br /><br />
+ [5] Brown, N.L., Stoyanov, J.V., Kidd, S.P. & Hobman, J.L., 2003. The MerR family of transcriptional regulators. FEMS Microbiology Reviews, 27(2-3), pp.145-163.
+ <br /><br />
+ [6] Phillips, C., Canalizo-Hernandez, M., Yidirim, A., Schatz, G.C., Mondragon, A. & O'Halloran, T.V., 2015. Allosteric transcription regulation via changes in the overall topology of the core promoter. Science, 349(6250), pp.877-881.
+ <br /><br />
+ [7] Wang, Z.K., Gong, J.S., Su, C., et al., 2024. Multilevel systematic optimization to achieve efficient integrated expression of Escherichia coli. ACS Synthetic Biology, 13(9), pp.2887-2898. doi:10.1021/acssynbio.4c00280.
+ </span>
+
+
+
+
</div>
</div>
--
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