From ced4b2c8275d3b97194ec448002c896aab62ee9a Mon Sep 17 00:00:00 2001
From: Ruby Chang <ruby931109@gmail.com>
Date: Wed, 28 Sep 2022 22:15:22 +0800
Subject: [PATCH] modified: static/style.css modified:
wiki/pages/attributions.html modified: wiki/pages/engineering.html
modified: wiki/pages/part-collection.html modified:
wiki/pages/team.html
---
static/style.css | 101 +++-
wiki/pages/attributions.html | 300 +++++++++--
wiki/pages/engineering.html | 4 +-
wiki/pages/part-collection.html | 881 ++------------------------------
wiki/pages/team.html | 41 +-
5 files changed, 432 insertions(+), 895 deletions(-)
diff --git a/static/style.css b/static/style.css
index 43cd9b3..0cfedb8 100644
--- a/static/style.css
+++ b/static/style.css
@@ -650,11 +650,32 @@ html {
background-color: whitesmoke;
}
.temp_pic_advisor {
- width: 60%;
- height: 60%;
+ width: 40%;
+ height: 40%;
+ border-radius: 50%;
+ background-color: whitesmoke;
+}
+.team_advisor_photo_wrap {
+ width: 80%;
+ height: 100%;
border-radius: 50%;
background-color: whitesmoke;
}
+.advisor_photo {
+ display: flex;
+ flex-direction: column;
+ align-items: center;
+ justify-content: center;
+}
+.advisor_image_wrap {
+ display: flex;
+ width: 85%;
+ align-self: center;
+ flex-direction: row;
+ margin-bottom: 50px;
+ justify-content: space-between;
+ align-items: center;
+}
.teamtitleimage {
margin-top: 30px;
margin-bottom: 30px;
@@ -719,6 +740,12 @@ html {
border-radius: 50%;
background-color: #f6f6f6;
}
+.advisor_name {
+ color: #082076;
+ font-size: 30px;
+ text-align: center;
+ margin-top: 50px;
+}
.image_herosection {
/* height: 500px; */
overflow: hidden;
@@ -810,6 +837,7 @@ html {
align-items: center;
justify-content: center;
width: 100%;
+ margin-bottom: 50px;
}
.banner_header {
@@ -869,6 +897,9 @@ html {
.herosectionteam {
height: 55vw;
}
+ .advisor_image_wrap {
+ width: 90%;
+ }
.shadow {
height: 100%;
}
@@ -876,10 +907,7 @@ html {
margin-top: -20%;
}
.temp_pic_advisor {
- width: 90%;
- height: 90%;
- border-radius: 50%;
- background-color: whitesmoke;
+ display: none;
}
.teamtitleword {
font-size: 60px;
@@ -938,11 +966,35 @@ html {
.attribubtion_wrap {
width: 80%;
margin-top: 200px;
+ margin-bottom: 200px;
display: flex;
flex-direction: column;
align-items: center;
justify-content: center;
}
+.jobtitleswrap {
+ display: flex;
+ flex-direction: row;
+}
+.job_title {
+ color: #91cdd1;
+ font-size: 28px;
+}
+.team_membername {
+ color: #676769;
+ font-size: 28px;
+ margin-left: 20px;
+}
+.team_memberwrap {
+ margin-top: 15px;
+ align-self: flex-start;
+ margin-bottom: 15px;
+}
+.workdistribution {
+ font-size: 20px;
+ margin-top: 20px;
+ color: #2e2e30;
+}
.teamtitleimage {
width: 40%;
}
@@ -1088,6 +1140,13 @@ html {
height: 1px;
background-color: white;
}
+.team_divider {
+ width: 100%;
+ border: none;
+ outline: none;
+ height: 1px;
+ background-color: gray;
+}
#yellowoutreachparts {
background-color: #f1d48d;
}
@@ -1227,7 +1286,7 @@ html {
position: absolute;
top: 0;
padding: 30px;
- margin-top: 80px;
+ padding-top: 80px;
height: 100vh;
right: 0px;
overflow-y: auto;
@@ -1407,7 +1466,6 @@ html {
position: fixed;
top: 80px;
left: 0;
- z-index: 30000;
height: 4px;
width: var(--progress);
background-color: #082076;
@@ -1544,6 +1602,9 @@ html {
.chart_image {
height: 200px;
}
+.chart_part_image {
+ height: 100px;
+}
.chart_contents {
background-color: white;
@@ -1619,6 +1680,14 @@ html {
grid-template-rows: repeat(6, 1fr);
grid-template-columns: repeat(4, 1fr);
}
+#three_five {
+ grid-template-rows: repeat(5, 1fr);
+ grid-template-columns: repeat(3, 1fr);
+}
+#three_two {
+ grid-template-rows: repeat(2, 1fr);
+ grid-template-columns: repeat(3, 1fr);
+}
#five_eight {
grid-template-rows: repeat(8, 1fr);
grid-template-columns: repeat(5, 1fr);
@@ -1698,10 +1767,9 @@ html {
margin-bottom: 50px;
}
#safetyimage {
- margin: 0;
+ margin: 0;
height: 300px;
- margin-right: 10px;
- margin-left: 10px;
+ margin: 10px;
}
.constant_width {
height: 280px;
@@ -1751,6 +1819,9 @@ html {
font-size: 17px;
margin-top: 43px;
}
+.info_contents_parts {
+ font-size: 17px;
+}
.workscited {
width: 100%;
font-size: 25px;
@@ -1793,13 +1864,14 @@ html {
height: 200px;
}
.constant_width {
- height: 150px;
+ width: 100%;
+ height: auto;
}
}
@media screen and (max-width: 1130px) {
#experimentalcontents {
width: 100%;
- padding: 150px;
+ padding: 75px;
margin: 0;
}
.search_index {
@@ -1807,6 +1879,9 @@ html {
width: 0;
margin: 0;
}
+ .safety_image_wrap {
+ flex-direction: column;
+ }
}
/*contents*/
diff --git a/wiki/pages/attributions.html b/wiki/pages/attributions.html
index a03e9cd..c9e42b8 100644
--- a/wiki/pages/attributions.html
+++ b/wiki/pages/attributions.html
@@ -1,44 +1,269 @@
{% extends "layout.html" %} {% block page_content %}
-<body class="body">
- <div class="wrap">
- <div class="loadingwrap">
- <img
- class="loadinggif"
- src="https://static.igem.wiki/teams/4271/wiki/logo.gif"
- />
+<div class="wrap">
+ <div class="loadingwrap">
+ <img
+ class="loadinggif"
+ src="https://static.igem.wiki/teams/4271/wiki/logo.gif"
+ />
+ <img
+ class="loadingwords"
+ src="https://static.igem.wiki/teams/4271/wiki/loading2-0.gif"
+ />
+ </div>
+ <div class="bannerwrap">
+ <img
+ class="bannerimg"
+ src="https://static.igem.wiki/teams/4271/wiki/attribution-banner.png"
+ />
+ </div>
+ <div class="attribubtion_wrap">
+ <div class="attribution_specialthanks">
<img
- class="loadingwords"
- src="https://static.igem.wiki/teams/4271/wiki/loading2-0.gif"
+ class="specialthanks"
+ src="https://static.igem.wiki/teams/4271/wiki/attribution-specialthanks.png"
/>
+ <div class="specialthanks_words">
+ Eutro in Vitro is delighted to announce that after months of
+ brainstorming, research, and experiments, we have successfully developed
+ a ground-breaking and effective solution to eutrophication. However, we
+ could not possibly accomplish so much in such a short period of time
+ without the help of many, including Dr. Chi-Yu Chen, Prof. Po-Husn
+ Huang, Institute of Clinical Medicine, NYCU, which provided lab
+ supervision and lab equipment, BIOTOOLS and Abreal Biotech, which
+ provided material and chemicals for experimental uses. Words are not
+ enough to express our appreciation to everyone that accompanied us on
+ this journey. Apart from that, we also want to express our gratitude to
+ the welcoming scientific community that aided us in numerous
+ disciplines.
+ </div>
</div>
- <div class="bannerwrap">
- <img
- class="bannerimg"
- src="https://static.igem.wiki/teams/4271/wiki/attribution-banner.png"
- />
+
+ <b class="title_team">Work Distribution</b>
+ <div class="team_memberwrap">
+ <div class="jobtitleswrap">
+ <b class="job_title">Julie Lin</b>
+ <div class="team_membername">Wet lab</div>
+ </div>
+ <div class="workdistribution">
+ Experimental Design, Experimentation, Figures, OPH proof of concept
+ (data analysis & documentation)
+ </div>
</div>
- <div class="attribubtion_wrap">
- <div class="attribution_specialthanks">
- <img
- class="specialthanks"
- src="https://static.igem.wiki/teams/4271/wiki/attribution-specialthanks.png"
- />
- <div class="specialthanks_words">
- Eutro in Vitro is delighted to announce that after months of
- brainstorming, research, and experiments, we have successfully
- developed a ground-breaking and effective solution to eutrophication.
- However, we could not possibly accomplish so much in such a short
- period of time without the help of many, including NYCU laboratory,
- which provided lab supervision and lab equipemnt, BIOTOOLS and Abreal
- Biotech, which provided lab equipment and chemicals for experimental
- uses. Words are not enough to express our appreciation to everyone
- that accompanied us on this journey. Apart from that, we also want to
- express our gratitude to the welcoming scientific community that aided
- us in numerous disciplines.
- </div>
+ <hr class="team_divider" />
+ <div class="team_memberwrap">
+ <div class="jobtitleswrap">
+ <b class="job_title">Ryan Hsu</b>
+ <div class="team_membername">Wet lab</div>
</div>
+ <div class="workdistribution">
+ Experimental Design, Experimentation, Safety & security documentation,
+ Proposed Implementation Design
+ </div>
+ </div>
+ <hr class="team_divider" />
+ <div class="team_memberwrap">
+ <div class="jobtitleswrap">
+ <b class="job_title">Penny Ho</b>
+ <div class="team_membername">Wet lab</div>
+ </div>
+ <div class="workdistribution">
+ Experimental Design, Experimentation, Figures, Engineering Success
+ Documentation
+ </div>
+ </div>
- <b class="title_team">Outreach</b>
+ <hr class="team_divider" />
+ <div class="team_memberwrap">
+ <div class="jobtitleswrap">
+ <b class="job_title">Kathryn Tsai</b>
+ <div class="team_membername">Wet lab</div>
+ </div>
+ <div class="workdistribution">
+ Experimental Design, Experimentation, Figures, AsPhoU proof of concept
+ (data analysis & documentation)
+ </div>
+ </div>
+
+ <hr class="team_divider" />
+ <div class="team_memberwrap">
+ <div class="jobtitleswrap">
+ <b class="job_title">Ethan Ho</b>
+ <div class="team_membername">Wet lab</div>
+ </div>
+ <div class="workdistribution">
+ Experimental Design, Experimentation, Parts Documentation, Parts
+ Registry, Photography
+ </div>
+ </div>
+
+ <hr class="team_divider" />
+ <div class="team_memberwrap">
+ <div class="jobtitleswrap">
+ <b class="job_title">Abraham Lin</b>
+ <div class="team_membername">Dry lab</div>
+ </div>
+ <div class="workdistribution">
+ Model, implementation (Design), Hardware (3D printing)
+ </div>
+ </div>
+
+ <hr class="team_divider" />
+ <div class="team_memberwrap">
+ <div class="jobtitleswrap">
+ <b class="job_title">Jessie Chen</b>
+ <div class="team_membername">Dry lab</div>
+ </div>
+ <div class="workdistribution">
+ Implementation (Design), Hardware documentation, Prototype crafting
+ </div>
+ </div>
+
+ <hr class="team_divider" />
+ <div class="team_memberwrap">
+ <div class="jobtitleswrap">
+ <b class="job_title">Joseph Haung</b>
+ <div class="team_membername">Dry lab</div>
+ </div>
+ <div class="workdistribution">Implementation (Arduino)</div>
+ </div>
+
+ <hr class="team_divider" />
+ <div class="team_memberwrap">
+ <div class="jobtitleswrap">
+ <b class="job_title">Carol Kao</b>
+ <div class="team_membername">Human practice</div>
+ </div>
+ <div class="workdistribution">
+ Interview (questions design, outreach), Wiki website (design), Education
+ (lecture), Virtual meet-up
+ </div>
+ </div>
+
+ <hr class="team_divider" />
+ <div class="team_memberwrap">
+ <div class="jobtitleswrap">
+ <b class="job_title">Una Chuang</b>
+ <div class="team_membername">Human practice</div>
+ </div>
+ <div class="workdistribution">
+ Interview (questions design, outreach), Education (lecture), Virtual
+ meet-up
+ </div>
+ </div>
+ <hr class="team_divider" />
+ <div class="team_memberwrap">
+ <div class="jobtitleswrap">
+ <b class="job_title">Alyssa Yang</b>
+ <div class="team_membername">Human practice</div>
+ </div>
+ <div class="workdistribution">
+ Education (planning, lecture), Media Outreach, Podcast(brainstorm),
+ Virtual meet-up, Interview
+ </div>
+ </div>
+
+ <hr class="team_divider" />
+ <div class="team_memberwrap">
+ <div class="jobtitleswrap">
+ <b class="job_title">Vincent Chiu</b>
+ <div class="team_membername">Human practice</div>
+ </div>
+ <div class="workdistribution">
+ Education (lecture), Podcast (brainstorm, edit & update), Collaboration
+ (planning, contacting), Interview
+ </div>
+ </div>
+
+ <hr class="team_divider" />
+ <div class="team_memberwrap">
+ <div class="jobtitleswrap">
+ <b class="job_title">Phoebe Lee</b>
+ <div class="team_membername">Human practice</div>
+ </div>
+ <div class="workdistribution">
+ Education (lecture), social media, Virtual meet-up, Interview
+ </div>
+ </div>
+
+ <hr class="team_divider" />
+ <div class="team_memberwrap">
+ <div class="jobtitleswrap">
+ <b class="job_title">Ruby Chang</b>
+ <div class="team_membername">Wiki</div>
+ </div>
+ <div class="workdistribution">
+ Wiki website design, Wiki website coding
+ </div>
+ </div>
+
+ <hr class="team_divider" />
+ <div class="team_memberwrap">
+ <div class="jobtitleswrap">
+ <b class="job_title">Kyle Wang</b>
+ <div class="team_membername">Wiki</div>
+ </div>
+ <div class="workdistribution">
+ Software design, Software coding, Wiki website coding
+ </div>
+ </div>
+
+ <hr class="team_divider" />
+ <div class="team_memberwrap">
+ <div class="jobtitleswrap">
+ <b class="job_title">I-Fan Tu</b>
+ <div class="team_membername">PI</div>
+ </div>
+ <div class="workdistribution">
+ Synbio courses, lab training, content revise and editting, lab
+ supervision
+ </div>
+ </div>
+
+ <hr class="team_divider" />
+ <div class="team_memberwrap">
+ <div class="jobtitleswrap">
+ <b class="job_title">Yu Chuan Lee</b>
+ <div class="team_membername">Instructor</div>
+ </div>
+ <div class="workdistribution">
+ Advisory, content revise and editting, lab supervision
+ </div>
+ </div>
+
+ <hr class="team_divider" />
+ <div class="team_memberwrap">
+ <div class="jobtitleswrap">
+ <b class="job_title">Ting-Yi Wu</b>
+ <div class="team_membername">Instructor</div>
+ </div>
+ <div class="workdistribution">
+ Advisory, content editting
+ </div>
+ </div>
+
+ <hr class="team_divider" />
+ <div class="team_memberwrap">
+ <div class="jobtitleswrap">
+ <b class="job_title">Cheng-Ju Lu</b>
+ <div class="team_membername">Instructor</div>
+ </div>
+ <div class="workdistribution">
+ Modelling courses, Advisory, content revise and editting
+ </div>
+ </div>
+
+ <hr class="team_divider" />
+ <div class="team_memberwrap">
+ <div class="jobtitleswrap">
+ <b class="job_title">Wan-Ju Chen</b>
+ <div class="team_membername">Instructor</div>
+ </div>
+ <div class="workdistribution">
+ Advisory
+ </div>
+ </div>
+
+ <!-- <b class="title_team">Outreach</b>
<div class="outreachwrap">
<div class="outreach_wrap">
<div class="outreachparts" id="yellowoutreachparts">
@@ -51,7 +276,7 @@
<hr class="outreach_divider" />
<div class="outreach_content">Contents</div>
</div>
-
+
<div class="outreachparts" id="greenoutreachparts">
<div class="outreach_name">NAME</div>
<hr class="outreach_divider" />
@@ -87,9 +312,8 @@
<div class="sponsors_logo"></div>
<div class="sponsors_logo"></div>
</div>
- </div>
- </div>
+ </div> -->
</div>
-</body>
+</div>
{% endblock %}
diff --git a/wiki/pages/engineering.html b/wiki/pages/engineering.html
index cd22e11..81ab84b 100644
--- a/wiki/pages/engineering.html
+++ b/wiki/pages/engineering.html
@@ -932,7 +932,7 @@
class="content_link"
href="{{ url_for('pages', page='implementation') }}"
>
- Implementation page.
+ Implementation page
</a>
.
</div>
@@ -957,7 +957,7 @@
and generate false signals for detection. For more information on our
team’s partnership with NYCU_Taipei, please visit our
<a class="content_link" href="{{ url_for('pages', page='partnership') }}">
- Partnership page.
+ Partnership page
</a>
.
</div>
diff --git a/wiki/pages/part-collection.html b/wiki/pages/part-collection.html
index faaf2e3..576bffd 100644
--- a/wiki/pages/part-collection.html
+++ b/wiki/pages/part-collection.html
@@ -26,915 +26,122 @@
<div class="wiki_contents">
<div class="search_index_wrap">
<div class="search_index">
- <div class="search_subindex" onclick="movetoone()">Biosensor Design</div>
+ <div class="search_subindex" onclick="movetoone()">Basic Parts</div>
<div class="search_subindex" onclick="movetotwo()">
- Biosensor Function
- </div>
- <div class="search_subindex" onclick="movetothree()">
- Experimental Methods
- </div>
- <div class="search_subindex" onclick="movetofour()">
- Experimental Results, Learning, and Redesign
- </div>
- <div class="search_subindex" onclick="movetofive()">
- Bacterial System for Cellular Inorganic Phosphate (Pi) Concentration
- Regulation
- </div>
- <div class="search_subindex" onclick="movetosix()">
- Theoretical Function of AsPhoU
- </div>
- <div class="search_subindex" onclick="movetoseven()">
- Preliminary Experiment: 5-Bromo-4-chloro-3-indolyl phosphate (XP)
- Coloration Test of Pho Regulon
- </div>
- <div class="search_subindex" onclick="movetoeight()">
- Malachite Green Coloration
- </div>
- <div class="search_subindex" onclick="movetonine()">
- Theoretical Function of our Assay Kit
- </div>
- <div class="search_subindex" onclick="movetoten()">
- Future plan
+ Composite Parts
</div>
</div>
</div>
<div class="herowrap" id="experimentalcontents">
<div id="progress-bar"></div>
<b class="heading1" id="greenheading1">
- The Paraoxon Hydrolysis Detection Experiment
+ Parts Page
</b>
<div class="index_container"></div>
-
- <div class="info_contents" id="greencontents">
- Insecticides are one of the most significant sources of organic phosphate
- pollution in water bodies. As the active metabolite of organophosphate
- insecticide, paraoxon could result in neurotoxic poisoning in animals. Our
- implementation hardware included a filtering device that contains bacteria
- engineered to overexpress the
- <em>oph</em>
- gene, which encodes the enzyme organophosphate hydrolase (OPH).
- </div>
- <div class="info_contents" id="greencontents">
- In order to prove the ability of OPH to hydrolyze and, thus, detoxify
- paraoxon, we utilize the pNP sensor as the specified switch.
- </div>
- <div class="index_container" id="one"></div>
- <div class="heading2" id="greenheading2">Biosensor Design</div>
-
- <div class="info_contents" id="greencontents">
- To determine the effect of the paraoxon hydrolysis reaction, we detected
- the amount of the reaction’s product, p-nitrophenol (pNP) with the pNP
- sensor. We designed the biosensor by transforming the pNP sensor plasmid
- and enzyme plasmid into the biological system
- <em>E. coli</em>
- BL21(DE3). The enzyme plasmid contains the original lac operon and the
- subcloned
- <em>oph</em>
- gene. The sensor plasmid contains the subcloned GFP (green fluorescent
- protein) gene and pNP mut1-1 gene, which encodes for the transcription
- factor protein to enhance downstream GFP expression when induced by pNP
- (Jha et al. 8495).
- </div>
- <div class="index_container" id="two"></div>
- <div class="heading2" id="greenheading2">Biosensor Function</div>
- <div class="info_contents" id="greencontents">
- The biosensor functions fully under exposure to both IPTG and paraoxon in
- the environment. With the induction of IPTG, the repressor protein is
- prevented from binding to the gene, allowing the transcription of the OPH
- protein. OPH then hydrolyses paraoxon, producing p-nitrophenol (pNP),
- which would bind to the activator protein pNP mut1-1, leading to the
- increased transcription of the GFP protein and, thus, causing the emission
- of green fluorescence (Jha et al. 8495). By measuring the intensity of GFP
- fluorescence, we can detect the detoxification of paraoxon and the level
- of inorganic phosphate production.
- </div>
- <img
- class="constant_height"
- src="https://static.igem.wiki/teams/4271/wiki/paraoxon-biosensor.png "
- />
- <div class="image_description" id="greencontents">
- Fig. 1 The function of the whole-cell biosensor
- </div>
- <img
- class="constant_height"
- id="lower_height"
- src="https://static.igem.wiki/teams/4271/wiki/biosensor-construct.png"
- />
- <div class="image_description" id="greencontents">
- Fig. 2 The construct and function of the pNP sensor plasmid
- </div>
- <div class="index_container" id="three"></div>
- <div class="heading2" id="greenheading2">Experimental Methods</div>
- <em class="info_contents_italics" id="greencontents">
- Experimental work 1: pNP Titration
- </em>
- <div class="info_contents" id="greencontents">
- To determine the standard curves for the positive correlation between pNP
- and GFP fluorescence intensity for further experiments, we titrated E.
- coli BL21 (DE3) carrying pNP sensor with varying concentrations of pNP.
- The concentrations and the resulting fluorescence intensity are mapped out
- as the standard curve for experimental references.
- </div>
- <em class="info_contents_italics" id="greencontents">
- Experimental work 2: Paraoxon Hydrolysis
- </em>
- <div class="info_contents" id="greencontents">
- Experimental work 2: Paraoxon Hydrolysis
- <em>E. coli</em>
- BL21 (DE3) carrying pET22b::OPH in presence of IPTG is treated with
- paraoxon. The rate of paraoxon hydrolysis by OPH can be calculated from
- the pNP production to prove the target protein's effectiveness.
- </div>
-
- <div class="index_container" id="four"></div>
<div class="heading2" id="greenheading2">
- Experimental Results, Learning, and Redesign
- </div>
- <div class="info_contents" id="greencontents">
- The data of the preliminary experiment, which aimed to test the
- transcriptional factor induced by pNP, showed no significance. The results
- were inconsistent with the research paper on which the experimental design
- was based. For more information on the engineering of the pNP sensor gene,
- please visit the
- <a class="content_link" href="{{ url_for('pages', page='engineering') }}">
- Engineering Success page
- </a>
- .
- </div>
- <div class="chart" id="two_five">
- <b class="chart_header" id="greencontents">
- Groups
- </b>
- <b class="chart_header" id="greencontents">
- Fluorescence
- </b>
- <div class="chart_contents" id="greencontents">DH5alpha</div>
- <div class="chart_contents" id="greencontents">24870</div>
- <div class="chart_contents" id="greencontents">DH5alpha + pNP</div>
- <div class="chart_contents" id="greencontents">20650</div>
- <div class="chart_contents" id="greencontents">DH5alpha-sensor</div>
- <div class="chart_contents" id="greencontents">46867</div>
- <div class="chart_contents" id="greencontents">DH5alpha-sensor + pNP</div>
- <div class="chart_contents" id="greencontents">50783</div>
- </div>
- <img
- class="constant_height"
- src="https://static.igem.wiki/teams/4271/wiki/fluorescent-protein-expression-level.png"
- />
- <div class="image_description" id="greencontents">
- Fig. 3 Green fluorescence detection in
- <em>E. coli</em>
- BL21 (DE3) and pNP sensor cell in absence and presence of pNP.
+ Parts for Eutrophic Water Purification (OPH and AsPhoU)
</div>
- <div class="info_contents" id="greencontents">
- Therefore, we redesigned a more direct method to test the degradation of
- paraoxon by OPH. The experimental design was based on the hypothesis that
- paraoxon (transparent) would be hydrolyzed to produce diethyl phosphate
- (transparent) and pNP (yellow, absorbance peak at 410 nm by
- spectrophotometry). We detected the production level of pNP with
- spectrophotometry to test the percentage of paraoxon degradation by OPH
- that could hydrolyze in a certain period of time. We performed two
- experimental works: the amount of pNP detection at the various time points
- (pNP conc. v.s. Time) and in the presence of different IPTG concentrations
- at a fixed time (pNP conc. v.s. IPTG conc.), respectively, to determine
- the optimal factors of OPH degradation activity.
- </div>
- <em class="info_contents_italics" id="greencontents">
- Preliminary Experiment
+ <div class="index_container" id="one"></div>
+ <em class="info_contents_parts" id="greencontents">
+ Basic Parts
</em>
- <div class="info_contents" id="greencontents">
- To ensure the function of OPH before introducing other variables and
- performing complete experiments with all control groups, we designed and
- conducted a preliminary experiment. 3 groups of BL21 (DE3) bacteria and 3
- groups of BL21 (DE3) bacteria engineered with OPH gene are cultured with
- medium only as the negative control, paraoxon, and pNP, respectively. The
- engineered bacteria are then induced by IPTG for protein expression. To
- test if the products of the hydrolysis reaction would be released into the
- environment, we measured the samples from the bacterial suspension and
- supernatant after centrifugation. We then deducted the background data
- (negative control) from the absorbance and divided the result by the
- optical density of the bacteria to test the level of pNP produced (and,
- therefore, the paraoxon degraded) per 10^7 CFU of bacteria added,
- indicating the ability of paraoxon degradation by each bacteria.
- </div>
- <div class="chart" id="four_seven">
+ <div class="chart" id="three_five">
<b class="chart_header" id="greencontents">
- Groups
+ Part Number
</b>
<b class="chart_header" id="greencontents">
- (Absorbance at 410nm - background data) / OD600
+ Description
</b>
<b class="chart_header" id="greencontents">
- (Supernatant Absorbance at 410nm - background data) / OD600
+ Diagram
</b>
- <b class="chart_header" id="greencontents"></b>
- <div class="chart_contents" id="greencontents">
- 1. BL2(DE3) (negative control)
- </div>
- <div class="chart_contents" id="greencontents">0</div>
- <div class="chart_contents" id="greencontents">0</div>
<div class="chart_contents" id="greencontents">
- <img
- class="chart_image"
- src="https://static.igem.wiki/teams/4271/wiki/1.jpg "
- />
- </div>
- <div class="chart_contents_highlight" id="greencontents">
- 2. BL2(DE3) +paraoxon (experimental)
- </div>
- <div class="chart_contents_highlight" id="greencontents">
- 0.2323266987
- </div>
- <div class="chart_contents_highlight" id="greencontents">
- 0.3905284832
- </div>
- <div class="chart_contents_highlight" id="greencontents">
- <img
- class="chart_image"
- src="https://static.igem.wiki/teams/4271/wiki/3.jpg "
- />
+ BBa_K4271000
</div>
<div class="chart_contents" id="greencontents">
- 3. BL2(DE3) +pNP (positive control)
+ Organophosphate Hydrolase + his-tag (OPH)
</div>
- <div class="chart_contents" id="greencontents">8.905950096</div>
- <div class="chart_contents" id="greencontents">9.966890595</div>
<div class="chart_contents" id="greencontents">
<img
- class="chart_image"
- src="https://static.igem.wiki/teams/4271/wiki/5.jpg "
+ class="chart_part_image"
+ src="https://static.igem.wiki/teams/4271/wiki/oph-his-tag.png"
/>
</div>
<div class="chart_contents" id="greencontents">
- 4. PET::OPH +IPTG induction (negative control)
- </div>
- <div class="chart_contents" id="greencontents">0</div>
- <div class="chart_contents" id="greencontents">0</div>
- <div class="chart_contents" id="greencontents">
- <img
- class="chart_image"
- src="https://static.igem.wiki/teams/4271/wiki/2.jpg "
- />
- </div>
- <div class="chart_contents_highlight" id="greencontents">
- 5. PET::OPH +paraoxon +IPTG induction (experimental)
- </div>
- <div class="chart_contents_highlight" id="greencontents">6.720481928</div>
- <div class="chart_contents_highlight" id="greencontents">6.916144578</div>
- <div class="chart_contents_highlight" id="greencontents">
- <img
- class="chart_image"
- src="https://static.igem.wiki/teams/4271/wiki/4.jpg "
- />
+ BBa_K4271002
</div>
<div class="chart_contents" id="greencontents">
- 6. PET::OPH +pNP +IPTG induction (positive control)
+ Antisense-PhoU (AsPhoU)
</div>
- <div class="chart_contents" id="greencontents">11.83912249</div>
- <div class="chart_contents" id="greencontents">12.51005484</div>
<div class="chart_contents" id="greencontents">
<img
- class="chart_image"
- src="https://static.igem.wiki/teams/4271/wiki/6.jpg "
+ class="chart_part_image"
+ src="https://static.igem.wiki/teams/4271/wiki/asphou.png"
/>
</div>
- </div>
- <em class="info_contents_italics" id="greencontents">
- Preliminary Experiment Results and Interpretation
- </em>
-
- <div class="info_contents" id="greencontents">
- The absorbance of the paraoxon solution increased 6.720481928 A.U.
- (highlighted in green) after the reaction with BL21 (DE3) engineered with
- the
- <em>oph</em>
- gene, whereas that of the paraoxon solution reacted with BL21(DE3) only
- increased 0.2323266987 A.U. (highlighted in green). The significant
- increase in the production of pNP proves the ability of our target protein
- OPH to hydrolyze paraoxon. Based on the successful results of our
- preliminary experiment, we further designed experimental works 1 and 2 to
- investigate the optimal reaction time and IPTG induction level,
- respectively, that would allow OPH to reach its highest enzyme expression
- and activity.
- </div>
- <em class="info_contents_italics" id="greencontents">
- Standard Curve Preparation
- </em>
- <div class="info_contents" id="greencontents">
- After observing that reaction with bacteria alters the absorbance of pNP
- solution significantly, we constructed a standard curve that measures the
- absorbance at 410 nm versus the concentration of pNP solution cultured
- with 0.6 O.D. bacteria for 6 hours.
- </div>
- <img
- class="constant_height"
- id="lower_height"
- src="https://static.igem.wiki/teams/4271/wiki/pnp-bacteria-standard-curve-6hr.png
-"
- />
- <div class="image_description" id="greencontents">
- Fig. 4 Standard Curve (410 absorbances v.s. pNP concentration)
- </div>
-
- <em class="info_contents_italics" id="greencontents">
- Experimental Work 1: pNP concentration v.s. Time
- </em>
- <div class="info_contents" id="greencontents">
- We aimed to test the optimal reaction time for OPH enzyme activity with
- experimental work 1. All solutions are prepared with 500 μM PXN/pNP
- chemical, 100 μM IPTG, and 0.6 O.D . bacteria. The experimental groups are
- designed as below:
- </div>
- <div class="chart" id="four_seven">
- <b class="chart_header" id="greencontents">
- Groups
- </b>
- <b class="chart_header" id="greencontents">
- Bacteria Used
- </b>
- <b class="chart_header" id="greencontents">
- Substrates
- </b>
- <b class="chart_header" id="greencontents">
- IPTG Induction
- </b>
- <div class="chart_contents" id="greencontents">1 (positive control)</div>
- <div class="chart_contents_span" id="greencontents">BL21(DE3)</div>
- <div class="chart_contents" id="greencontents">pNP</div>
- <div class="chart_contents" id="greencontents">induced</div>
- <div class="chart_contents" id="greencontents">2 (negative control)</div>
- <div class="chart_contents" id="greencontents">PXN</div>
- <div class="chart_contents" id="greencontents">uninduced</div>
- <div class="chart_contents" id="greencontents">
- 3 (experimental group)
- </div>
- <div class="chart_contents" id="greencontents">PXN</div>
- <div class="chart_contents" id="greencontents">induced</div>
- <div class="chart_contents" id="greencontents">4 (positive control)</div>
- <div class="chart_contents_span_second" id="greencontents">
- BL21(DE3) engineered with OPH
- </div>
- <div class="chart_contents" id="greencontents">pNP</div>
- <div class="chart_contents" id="greencontents">induced</div>
- <div class="chart_contents" id="greencontents">5 (negative control)</div>
- <div class="chart_contents" id="greencontents">PXN</div>
- <div class="chart_contents" id="greencontents">uninduced</div>
- <div class="chart_contents" id="greencontents">
- 6 (experimental group)
- </div>
- <div class="chart_contents" id="greencontents">PXN</div>
- <div class="chart_contents" id="greencontents">induced</div>
- </div>
- <div class="info_contents" id="greencontents">
- The absorbance at the wavelength of 410 nm and 600 nm in each group was
- measured after 1 hr, 2 hrs, 3 hrs, 4 hrs, 5 hrs, 6 hrs, and 24 hrs of
- reaction to test the pNP production level and the concentration of
- bacteria over time.
- </div>
- <em class="info_contents_italics" id="greencontents">
- Experimental 1 Results and Interpretation
- </em>
- <img
- class="constant_height"
- src="https://static.igem.wiki/teams/4271/wiki/time-v-s-pnp.png
- "
- />
-
- <div class="image_description" id="greencontents">
- Fig. 5 Time (hr) v.s. pNP concentration (μM)
- </div>
-
- <div class="info_contents" id="greencontents">
- The result of experimental work 1 shows that pNP concentration increases
- rapidly in the first 5 hours and grows steadily afterward.
- </div>
-
- <em class="info_contents_italics" id="greencontents">
- Experimental Work 2: pNP concentration v.s. IPTG concentration
- </em>
-
- <div class="info_contents" id="greencontents">
- We aimed to determine the optimal concentration of IPTG induction for OPH
- enzyme expression with experimental work 2. Solutions are prepared with
- 500 μM PXN/pNP chemical, 0.6 O.D. bacteria, and various concentrations of
- IPTG. The experimental groups are designed as below:
- </div>
-
- <div class="chart" id="four_seven">
- <b class="chart_header" id="greencontents">
- Groups
- </b>
- <b class="chart_header" id="greencontents">
- Bacteria Used
- </b>
- <b class="chart_header" id="greencontents">
- Substrates
- </b>
- <b class="chart_header" id="greencontents">
- IPTG Induction (μM)
- </b>
- <div class="chart_contents" id="greencontents">1 (negative control)</div>
- <div class="chart_contents_span_third" id="greencontents">BL21(DE3)</div>
- <div class="chart_contents" id="greencontents">-</div>
- <div class="chart_contents" id="greencontents">0</div>
- <div class="chart_contents" id="greencontents">2 (positive control)</div>
- <div class="chart_contents" id="greencontents">pNP</div>
- <div class="chart_contents" id="greencontents">0</div>
- <div class="chart_contents" id="greencontents">
- 3 (experimental group)
- </div>
- <div class="chart_contents" id="greencontents">PXN</div>
- <div class="chart_contents" id="greencontents">0</div>
- <div class="chart_contents" id="greencontents">4 (negative control)</div>
- <div class="chart_contents_span_four" id="greencontents">
- BL21(DE3) engineered with OPH
- </div>
- <div class="chart_contents" id="greencontents">-</div>
- <div class="chart_contents" id="greencontents">0</div>
- <div class="chart_contents" id="greencontents">5 (positive group)</div>
- <div class="chart_contents" id="greencontents">pNP</div>
- <div class="chart_contents" id="greencontents">0</div>
- <div class="chart_contents" id="greencontents">
- 6 (experimental group)
- </div>
- <div class="chart_contents" id="greencontents">PXN</div>
- <div class="chart_contents" id="greencontents">0</div>
<div class="chart_contents" id="greencontents">
- 7 (experimental group)
+ BBa_K4271015
</div>
- <div class="chart_contents" id="greencontents">PXN</div>
- <div class="chart_contents" id="greencontents">2000</div>
<div class="chart_contents" id="greencontents">
- 8 (experimental group)
+ pelB signal peptide
</div>
- <div class="chart_contents" id="greencontents">PXN</div>
- <div class="chart_contents" id="greencontents">1000</div>
<div class="chart_contents" id="greencontents">
- 9 (experimental group)
- </div>
- <div class="chart_contents" id="greencontents">PXN</div>
- <div class="chart_contents" id="greencontents">500</div>
- <div class="chart_contents" id="greencontents">
- 10 (experimental group)
- </div>
- <div class="chart_contents" id="greencontents">PXN</div>
- <div class="chart_contents" id="greencontents">250</div>
- <div class="chart_contents" id="greencontents">
- 11 (experimental group)
+ <img
+ class="chart_part_image"
+ src="https://static.igem.wiki/teams/4271/wiki/pelb-signal-peptide.png"
+ />
</div>
- <div class="chart_contents" id="greencontents">PXN</div>
- <div class="chart_contents" id="greencontents">125</div>
<div class="chart_contents" id="greencontents">
- 12 (experimental group)
+ BBa_K4271016
</div>
- <div class="chart_contents" id="greencontents">PXN</div>
- <div class="chart_contents" id="greencontents">62.5</div>
<div class="chart_contents" id="greencontents">
- 13 (experimental group)
+ AsPhoU RBS
</div>
- <div class="chart_contents" id="greencontents">PXN</div>
- <div class="chart_contents" id="greencontents">31.25</div>
<div class="chart_contents" id="greencontents">
- 14 (experimental group)
+ <img
+ class="chart_part_image"
+ src="https://static.igem.wiki/teams/4271/wiki/pelb-signal-peptide.png"
+ />
</div>
- <div class="chart_contents" id="greencontents">PXN</div>
- <div class="chart_contents" id="greencontents">15.625</div>
- </div>
-
- <div class="info_contents" id="greencontents">
- The absorbance values at the wavelength of 410 nm and 600 nm were measured
- after 6 hrs and 23 hrs of reaction to test the pNP production level and
- the concentration of bacteria with increasing concentration of IPTG.
- </div>
- <em class="info_contents_italics" id="greencontents">
- Experimental Work 2 Results and Interpretation
- </em>
- <img
- class="constant_height"
- src="https://static.igem.wiki/teams/4271/wiki/iptg-v-s-pnp.png"
- />
-
- <div class="image_description" id="greencontents">
- Fig. 6 IPTG induction (μM) vs. pNP concentration after 23 hours (μM)
- </div>
- <div class="info_contents" id="greencontents">
- The results of experimental work 2 demonstrated that after 23 hours, the
- experimental group induced by 250 μM IPTG reached the highest level of pNP
- concentration, thus proving that 250 μM IPTG is the optimal induction for
- the highest OPH protein expression.
- </div>
-
- <b class="heading1" id="greenheading1">
- Inorganic Phosphate Detection Experiments
- </b>
- <div class="index_container"></div>
-
- <div class="info_contents" id="greencontents">
- Inorganic phosphate (Pi) accumulated in the sediments of aquatic
- environments is the primary reason for eutrophication. In order to
- decrease the amount of inorganic phosphate in water bodies, our team
- implemented a device that contains specially engineered
- <em>E. coli</em>
- that expresses the RNA of antisense PhoU (AsPhoU) thus hindering the
- expression of PhoU and enhancing phosphate transportation. Through our
- engineering process, these bacterial bodies can absorb more Pi from the
- environment.
- </div>
-
- <div class="index_container" id="five"></div>
- <div class="heading2" id="greenheading2">
- Bacterial System for Cellular Inorganic Phosphate (Pi) Concentration
- Regulation
- </div>
- <div class="info_contents" id="greencontents">
- PstSCAB is a high-affinity phosphate transporter protein in
- <em>E. coli</em>
- . Pho regulon is a common bacterial regulatory system determining and
- managing intracellular Pi concentration through the regulation of the
- PstSCAB transporter protein. The regulon consists of a histidine kinase
- sensor protein – PhoR – on the inner membrane and a response regulator –
- PhoB – on the cytoplasmic side of the membrane of a prokaryotic cell. The
- way PhoB regulates the simultaneous expression of PstSCAB and PhoA depends
- on the process by which PhoU binds onto PhoR. PhoU, a metal-binding
- protein, detects the intracellular concentration levels of Pi. When the
- environment undergoes a decrease in Pi concentration, PhoU dissociates
- from PhoR and the PstSCAB transporter. This, in turn, promotes the
- transportation of Pi and increases PhoA expression, which absorbs more Pi
- into the cell and enhances the phosphorylation of PhoB. On the other hand,
- when the phosphate level is high, PhoU would be activated to block its
- absorption, meaning that the transportation by the PstSCAB protein and the
- expression of PhoA would be inhibited.
- </div>
-
- <img
- class="constant_height"
- src="https://static.igem.wiki/teams/4271/wiki/pstscab.png "
- />
- <div class="image_description" id="greencontents">
- Fig. 1 Phosphate limitation; PstSCAB active; PhoR kinase inactive (left)
- and Phosphate sufficiency; PstSCAB inactive; PhoR kinase active (right)
- (Devine)
- </div>
-
- <div class="index_container" id="six"></div>
- <div class="heading2" id="greenheading2">
- Theoretical Function of AsPhoU
- </div>
- <div class="info_contents" id="greencontents">
- A previously published paper demonstrates that PhoU knockout leads to a
- halt
- <em>E. coli</em>
- growth (Haldimann). Therefore, we designed the mRNA for AsPhoU hindering
- phoU expression to increase the amount of Pi that bacteria could
- transport, thus lowering the concentration of Pi in aquatic environments.
- AsPhoU cell expresses antisense PhoU RNA under arabinose-control promoter
- (pBAD) and thus binds to the mRNA of PhoU, hindering ribosome binding to
- decrease phoU translation. The inhibition of PhoU protein would allow the
- PstSCAB transporter to be open for Pi transportation at all times, even
- under the high concentration of phosphate in eutrophic water bodies.
- </div>
- <img
- class="constant_height"
- src="https://static.igem.wiki/teams/4271/wiki/phou.png"
- />
- <div class="image_description" id="greencontents">
- Fig. 2 Normal PhoU protein function (left) and the inhibition of PhoU by
- AsPhoU, as well as its downstream effects (right)
- </div>
- <div class="info_contents" id="greencontents">
- With the purpose of proving the effectiveness of our engineered
- <em>E. coli</em>
- in decreasing the concentration of inorganic phosphate (Pi) in the
- environment, 5-Bromo-4-chloro-3-indolyl phosphate coloration and malachite
- green coloration are used.
- </div>
-
- <div class="index_container" id="seven"></div>
- <div class="heading2" id="greenheading2">
- Preliminary Experiment: 5-Bromo-4-chloro-3-indolyl phosphate (XP)
- Coloration Test of Pho Regulon
</div>
- <em class="info_contents_italics" id="greencontents">
- Experimental Design
- </em>
- <div class="info_contents" id="greencontents">
- To ensure the effectiveness and practicality of our aforementioned
- experimental design, we conducted a preliminary experiment.
- 5-Bromo-4-chloro-3-indolyl phosphate (XP) is a chromogenic substrate that
- shows no color in its stable state. However, if XP is in contact with
- PhoA, PhoA will hydrolyze XP, thus severing phosphate ions (a phosphate
- monoester + water = 5,5′-dibromo-4,4′-dichloro-indigo + a phosphate ion),
- and the remaining chemical will appear to be blue. Once an increase in the
- concentration of Pi is detected, the expression of PhoA rises and turns
- the XP solution blue. To test the function of our bacteria engineered with
- AsPhoU, we placed it inside the XP chemical to determine the absorbance of
- Pi by observing the color changes.
- </div>
- <em class="info_contents_italics" id="greencontents">
- Experimental Results and Interpretation
+ <div class="index_container" id="two"></div>
+ <em class="info_contents_parts" id="greencontents">
+ Composite Parts
</em>
- <div class="info_contents" id="greencontents">
- Our positive control group is
- <em>E. coli</em>
- DH5α in a low phosphate medium, which turns the XP solution blue,
- reflecting low phosphate to induce PhoA expression. On the contrary, no
- blue color was observed in a high phosphate medium.
- </div>
- <div class="chart" id="four_six">
+ <div class="chart" id="three_two">
<b class="chart_header" id="greencontents">
- Groups
+ Part Number
</b>
<b class="chart_header" id="greencontents">
- Environmental condition
+ Description
</b>
<b class="chart_header" id="greencontents">
- Resulting coloration of
- <em>E. coli</em>
- colonies
+ Diagram
</b>
- <b class="chart_header" id="greencontents"></b>
- <div class="chart_contents" id="greencontents">
- <em>E. coli</em>
- DH5α
- </div>
- <div class="chart_contents" id="greencontents">Low phosphate</div>
- <div class="chart_contents" id="greencontents">Blue</div>
- <div class="chart_contents" id="greencontents">
- <img
- class="chart_image"
- src="https://static.igem.wiki/teams/4271/wiki/e-coli-low-p.png"
- />
- </div>
- <div class="chart_contents" id="greencontents">
- <em>E. coli</em>
- DH5α (withAsPhoU)
- </div>
- <div class="chart_contents" id="greencontents">
- Low phosphate
- </div>
- <div class="chart_contents" id="greencontents">
- Blue
- </div>
- <div class="chart_contents" id="greencontents">
- <img
- class="chart_image"
- src="https://static.igem.wiki/teams/4271/wiki/e-coli-asphou-low-p.png"
- />
- </div>
<div class="chart_contents" id="greencontents">
- <em>E. coli</em>
- DH5α (with AsPhoU) + arabinose
+ BBa_K4271001
</div>
- <div class="chart_contents" id="greencontents">Low phosphate</div>
- <div class="chart_contents" id="greencontents">Blue</div>
<div class="chart_contents" id="greencontents">
- <img
- class="chart_image"
- src="https://static.igem.wiki/teams/4271/wiki/e-coli-asphou-arabinose-low-p.png"
- />
+ T7 Promoter + Lac operator + RBS + pelB + OPH + his-tag + T7 terminator
</div>
- <div class="chart_contents" id="greencontents">
- <em>E. coli</em>
- DH5α
- </div>
- <div class="chart_contents" id="greencontents">High phosphate</div>
- <div class="chart_contents" id="greencontents">Transparent</div>
<div class="chart_contents" id="greencontents">
<img
class="chart_image"
- src="https://static.igem.wiki/teams/4271/wiki/e-coli-high-p.png"
+ src="https://static.igem.wiki/teams/4271/wiki/linear-map-pet22boph.jpg"
/>
</div>
<div class="chart_contents" id="greencontents">
- <em>E. coli</em>
- DH5α (with AsPhoU)
+ BBa_K4271003
</div>
<div class="chart_contents" id="greencontents">
- High phosphate
+ araBAD promoter + RBS + AsPhoU + T1 T2 terminator
</div>
- <div class="chart_contents" id="greencontents">Transparent</div>
<div class="chart_contents" id="greencontents">
<img
class="chart_image"
- src="https://static.igem.wiki/teams/4271/wiki/e-coli-asphou-high-p.png"
+ src="https://static.igem.wiki/teams/4271/wiki/asphou-linear-map.png"
/>
</div>
- <div class="chart_contents_highlight" id="greencontents">
- <em>E. coli</em>
- DH5α (with AsPhoU) + arabinose
- </div>
- <div class="chart_contents_highlight" id="greencontents">
- High phosphate
- </div>
- <div class="chart_contents_highlight" id="greencontents">Blue</div>
- <div class="chart_contents_highlight" id="greencontents">
- <img
- class="chart_image"
- src="https://static.igem.wiki/teams/4271/wiki/e-coli-asphou-arabinose-high-p.png"
- />
- </div>
- </div>
- <div class="info_contents" id="greencontents">
- After the addition of
- <em>E. coli</em>
- DH5α (with AsPhoU) with 0.2 % arabinose at a high phosphate concentration,
- the XP solution turned blue, proving the expression of PhoA and, thus, the
- transportation of Pi into the bacterial cells.
- </div>
-
- <div class="index_container" id="eight"></div>
- <div class="heading2" id="greenheading2">Malachite Green Coloration</div>
-
- <em class="info_contents_italics" id="greencontents">
- Experimental Design
- </em>
-
- <div class="info_contents" id="greencontents">
- Once molybdate (MoOâ‚„â»Â²) comes into contact with phosphate (POâ‚„â»Â³), a
- complex containing phosphomolybdic acid would usually form. This complex
- interacts with malachite, forming a green chromogenic complex that remains
- in its most stable phase in an acidic environment. The color formation
- runs proportional to the increase of phosphate concentration. The shifts
- in color could determine the absorbance at 620 nm by a spectrophotometer.
- We would add the aforementioned chemical into our bacteria to interpret
- the effectiveness of our engineering method.
- </div>
- <em class="info_contents_italics" id="greencontents">
- Standard Curve Preparation
- </em>
-
- <div class="info_contents" id="greencontents">
- Our team constructed a standard curve that measures the absorbance at 620
- nm versus the concentration of phosphate solution in the presence of E.
- coli of 0.1 O.D.
- </div>
- <img
- class="constant_height"
- src="https://static.igem.wiki/teams/4271/wiki/standard-curve.png "
- />
- <div class="image_description" id="greencontents">
- Fig. 3 Standard Curve (​​620 nm absorbance vs phosphate conc.)
- </div>
- <em class="info_contents_italics" id="greencontents">
- Experimental Work 1: Time versus Pi Concentration
- </em>
- <div class="info_contents" id="greencontents">
- We aimed to test the optimal time of our engineered
- <em>E. coli</em>
- functioning under different conditions. All simulated high phosphate
- solutions are prepared with 2 mM of Kâ‚‚HPOâ‚„, 0.06% glucose, and MOPS
- buffer; all simulated low phosphate solutions are prepared with 0.1 mM of
- Kâ‚‚HPOâ‚„, 0.4% glucose, and MOPS buffer.
- </div>
- <div class="info_contents" id="greencontents">
- We incubated our engineered
- <em>E. coli</em>
- (cultured at 0.1 O.D.) under the same condition, added malachite and
- molybdate into our
- <em>E. coli</em>
- colonies, and detected the absorbance at 600 and 620 nm, respectively,
- reflecting cell density and inorganic phosphate (Pi) concentration at
- different time points. Using the standard curve above, we translated the
- detected data of absorbance at 620 nm into phosphate concentration in μM.
- </div>
- <em class="info_contents_italics" id="greencontents">
- Experimental Results and Interpretation
- </em>
- <div class="info_contents" id="greencontents">
- Experiments on the bacterial body containing AsPhoU in the presence or
- absence of arabinose were conducted for the level of phosphate absorption.
- The groups with arabinose induction clearly reported higher absorbed
- phosphate concentrations as AsPhoU is expressed. More phosphate absorption
- in presence of arabinose was observed compared to the absence of arabinose
- in AsPhoU. The maximal amount of Pi absorption was 3.1 x 10-6 μM per CFU
- (colony forming unit) for 1-hour culture, estimating 2.52 mM phosphate
- absorption per OD.
- </div>
- <div class="info_contents" id="greencontents">
- The full set experimental bar graph (1, 2, 3 hrs) indicating efficiency
- are illustrated below:
- </div>
- <img
- class="constant_height"
- src="https://static.igem.wiki/teams/4271/wiki/pi-content-per-cfu-versus-different-experimental-sets.png"
- />
- <div class="image_description" id="greencontents">
- Fig. 4 Phosphate content per CFU versus different experimental sets
- </div>
- <b class="heading1" id="greenheading1">
- Polyphosphate (PolyP) Detection Experiments
- </b>
- <div class="index_container"></div>
-
- <div class="info_contents" id="greencontents">
- Concluding from the previous experiments, AsPhoU cell has proven its
- effectiveness in absorbing a higher amount of Pi in the environment. While
- acknowledging that, we simultaneously have to make sure that the absorbed
- Pi constructively fixates into inorganic polyphosphate (polyP) so that it
- remains inside the bacterial bodies. Henceforth our team utilized
- Sigma-Aldrich’s PolyP assay kit, which helped in quantifying the amount of
- PolyP in bacterial bodies.
- </div>
-
- <div class="index_container" id="nine"></div>
- <div class="heading2" id="greenheading2">
- Theoretical Function of our Assay Kit
- </div>
- <div class="info_contents" id="greencontents">
- PolyP reacts fully with the fluorescent dye provided by the assay kit,
- forming a detectable complex. The fluorescence of the created complex is
- then measured with a spectrophotometer at λ (excitation) = 415 nm and λ
- (emission) = 550 nm. The amount of fluorescence detected runs proportional
- to the concentration of PolyP in the sampled bacteria.
- </div>
- <em class="info_contents_italics" id="greencontents">
- Standard Curve Preparation
- </em>
- <div class="info_contents" id="greencontents">
- Our team constructed a standard curve with the diluted PolyP solution and
- our assay buffer. The experimental groups are listed below:
- </div>
- <div class="chart" id="four_seven">
- <b class="chart_header" id="greencontents">
- Well
- </b>
- <b class="chart_header" id="greencontents">
- 10 uM Standard PolyP
- </b>
- <b class="chart_header" id="greencontents">
- PolyP Assay Buffer
- </b>
- <b class="chart_header" id="greencontents">
- PolyP (pmol/well)
- </b>
- <div class="chart_contents" id="greencontents">1</div>
- <div class="chart_contents" id="greencontents">0 uL</div>
- <div class="chart_contents" id="greencontents">50 uL</div>
- <div class="chart_contents" id="greencontents">0</div>
-
- <div class="chart_contents" id="greencontents">2</div>
- <div class="chart_contents" id="greencontents">5 uL</div>
- <div class="chart_contents" id="greencontents">45 uL</div>
- <div class="chart_contents" id="greencontents">50</div>
-
- <div class="chart_contents" id="greencontents">3</div>
- <div class="chart_contents" id="greencontents">10 uL</div>
- <div class="chart_contents" id="greencontents">40 uL</div>
- <div class="chart_contents" id="greencontents">100</div>
-
- <div class="chart_contents" id="greencontents">4</div>
- <div class="chart_contents" id="greencontents">14 uL</div>
- <div class="chart_contents" id="greencontents">35 uL</div>
- <div class="chart_contents" id="greencontents">150</div>
-
- <div class="chart_contents" id="greencontents">5</div>
- <div class="chart_contents" id="greencontents">20 uL</div>
- <div class="chart_contents" id="greencontents">30 uL</div>
- <div class="chart_contents" id="greencontents">200</div>
-
- <div class="chart_contents" id="greencontents">6</div>
- <div class="chart_contents" id="greencontents">25 uL</div>
- <div class="chart_contents" id="greencontents">25 uL</div>
- <div class="chart_contents" id="greencontents">250</div>
- </div>
- <img
- class="constant_height"
- src="https://static.igem.wiki/teams/4271/wiki/standard-curve-polyp-conc-vs-fluorescence-intensity.png"
- />
- <div class="image_description" id="greencontents">
- Fig. 5 Standard Curve (PolyP conc. vs. fluorescence intensity)
- </div>
-
- <div class="index_container" id="ten"></div>
- <div class="heading2" id="greenheading2">
- Future plan
- </div>
- <div class="info_contents" id="greencontents">
- Owing to time constraints, we have not been able to test the ability of
- AsPhoU for intracellular PolyP accumulation. Regardless, we would conduct
- this experimental process in the future, with the hope of reiterating that
- our proposed solution successfully operates. Moreover, we would subclone
- oph and AsPhoU to pACYCDuet vector with p15A ori, compatible with PolyP
- sensor carrying pMB ori to generate our PolyP sensor cell in this project.
- </div>
- <div class="workscited" id="greencontents">References</div>
- <div class="info_contents" id="greencontents">
- Jha, Ramesh K., et al. “A Microbial Sensor for Organophosphate Hydrolysis
- Exploiting an Engineered Specificity Switch in a Transcription Factor.â€
- Nucleic Acids Research, vol. 44, no. 17, 2016, pp. 8490–500. Crossref,
- https://doi.org/10.1093/nar/gkw687.
- </div>
- <div class="info_contents" id="greencontents">
- Devine, Kevin M. “Activation of the PhoPR-Mediated Response to Phosphate
- Limitation Is Regulated by Wall Teichoic Acid Metabolism in Bacillus
- subtilis.†Frontiers in microbiology vol. 9 2678. 6 Nov. 2018,
- doi:10.3389/fmicb.2018.02678
- </div>
- <div class="info_contents" id="greencontents">
- Haldimann, A et al. “Use of New Methods for Construction of Tightly
- Regulated Arabinose and Rhamnose Promoter Fusions in Studies of the
- Escherichia coli Phosphate Regulon.†Journal of bacteriology vol. 180,5
- (1998): 1277-86. doi:10.1128/JB.180.5.1277-1286.1998
</div>
</div>
</div>
diff --git a/wiki/pages/team.html b/wiki/pages/team.html
index ff59d9f..e80bc5b 100644
--- a/wiki/pages/team.html
+++ b/wiki/pages/team.html
@@ -1,6 +1,5 @@
{% extends "layout.html" %}
{% block page_content %}
-<body class="body">
<div class="wrap">
<div class="loadingwrap">
<img class="loadinggif" src="https://static.igem.wiki/teams/4271/wiki/logo.gif">
@@ -12,10 +11,10 @@
</div>
<div class="teamwrap">
- <b class="title_team">Team Advisors</b>
+ <b class="title_team">Team Instructor</b>
<div class="advisor-wrap">
- <div class="temp_pic_advisor">
+ <div class="temp_pic_advisor">
<img
class="image_advisor"
src="https://static.igem.wiki/teams/4271/wiki/advisor.png"
@@ -26,9 +25,42 @@
<div class="description_advisor">
"Working with young scientists always generates unexpected, unpredictable and cool ideas. "
</div>
+ </div>
+ </div>
+
+ <b class="title_team">Team Advisors</b>
+ <div class="advisor_image_wrap">
+ <div class="advisor_photo">
+ <div class="team_advisor_photo_wrap">
+ <img
+ class="image_advisor"
+ src="https://static.igem.wiki/teams/4271/wiki/advisor1.png"
+ />
+ </div>
+ <div class="advisor_name">Jenner</div>
+ </div>
+
+ <div class="advisor_photo">
+ <div class="team_advisor_photo_wrap">
+ <img
+ class="image_advisor"
+ src="https://static.igem.wiki/teams/4271/wiki/advisor2.png"
+ />
</div>
+ <div class="advisor_name">Danny</div>
+ </div>
+ <div class="advisor_photo">
+ <div class="team_advisor_photo_wrap">
+ <img
+ class="image_advisor"
+ src="https://static.igem.wiki/teams/4271/wiki/claire.png"
+ />
</div>
- <b class="title_team">Team Members</b>
+ <div class="advisor_name">Claire</div>
+ </div>
+
+ </div>
+ <b class="title_team">Team Members</b>
<div class="team-wrap">
<div class="person">
<div class="temp_pic">
@@ -381,5 +413,4 @@
</div>
</div>
</div>
-</body>
{% endblock %}
--
GitLab