Latest revision as of 10:03, 28 October 2013

Abstract

Hypertension has become the leading risk factor for mortality worldwide. Human’s blood pressure (BP) has a basic daily rhythm with two peaks,6:00 to 10:00 in the morning and 16:00 to18:00 in the afternoon. The morning peak is also called “death time”,for it’s unlikely to take drugs before waking. Propionate, a short chain fatty acid, was recently shown to produce an acute hypotensive response.2013 HUST-China iGEM use a synthetic way to combine bio-oscillator with propionate-producing enzyme gene, trying to build a gut probiotic which can release propionate periodically in accord with the rhythm of human BP. This could be a great substitute for chemical drugs by saving patients from drug dependence and the risk of sudden death at morning BP peak time.

Oscillator

The key part of the oscillator is araBAD/ lacZYA hybrid p- romoter. It is activated by the AraC protein in the prese- nce of arabinose and repressed by the LacI protein in the absence of IPTG, constructing two feedback loops with opposite effects.And the differential activity of the two feedback loops can drive oscillatory behaviour.

Propionate

Reactor

A four-gene operon in E.coli K12 genome which includes sbm,ygfG,ygfH and ygfD, is significant in the metabolic pathway that converts succinate to propionate through Wood-Werkman reaction. We constructed effective expression plasmid to increase the quantity of the four enzymes independently. By measuring the propionate amount, we figured out which of the four is the most effective.

Future work

In the near future, we will regulate the period of propionate utilizing the frequency divider with a ssrA-tag analog attached to the end of enzyme.Besides,we will replace mRFP with key gene in the synthetic pathway, hoping to see periodical release of propionate in accord with the rhythm of human BP.

Protocol

Our project is divided into three parts: the construction of the biological oscillator, the output evaluation of propionate by HPLC and the standardization of four genes.

Modeling

Detailed analysis of the oscillator makes us clearer about how it works. From the establishment of DDEs to parameter sweep and sensitivity analysis, we know how each parameter contributes to the period. The MCOS shows us the feasibility of a group of oscillators that can be eventually applied in practice in vivo (colon).Lastly, we statistically analyzed our simulated data and wet-lab data.

Human

Practice

We have done a remarkable job in introducing high school students synthetic biology and iGEM jamborees as well as motivating them towards future participation in the iGEM. Besides, we collaborated with two other iGEM teams by sharing plasmids and characterizing their parts. Furthermore, we made public speeches in our school about what we achieved,shared the felt in the iGEM competition.

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+<div style="float:left;"><a href="https://2013.igem.org" title="2013 IGEM Main page"><img src="https://static.igem.org/mediawiki/2013/4/4e/UNIK_Copenhagen_iGEM_logo.png" style="width:50px;"></a></div>
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-<div class="row" style="width:900px;height:250px;margin:20px 0 30px 50px;border:1px solid #fff;">
+<div><img src="https://static.igem.org/mediawiki/2013/3/3a/HUST-zzoscillating-BP-reliever.png" /></div>
+<div class="row" id="abstract" style="width:900px;height:250px;margin:20px 0 30px 50px;border:1px solid #fff;box-shadow:1px 1px 15px 5px rgba(0, 0, 0, 0.5)">
 				<div style="float:left;width:240px;text-align:center;margin-top:105px;"><h1>Abstract</h1></div>
 				<div style="float:left;width:600px;text-align:left;margin:30px 30px 0px;">
-					Hypertension has become the leading risk factor for mortality worldwide. Human’s blood pressure has a basic daily rhythm with two peaks,
+					Hypertension has become the leading risk factor for mortality worldwide. Human’s blood pressure (BP) has a basic daily rhythm with two
-:00 to 10:00 in the morn-
+                                        peaks,6:00 to 10:00 in the
-					ing and 16:00 to18:00 in the afternoon.The
+					morning and 16:00 to18:00 in the afternoon.
-					morning peak is also called “death tim-
+                                        The morning peak is also called “death
-					e”,for it’s unlikely to take drugs before waking. Propionate, a short chain fatty acid,
+					time”,for it’s unlikely to take drugs before waking. Propionate, a short chain fatty acid,
-					 was recently shown to produce an acute hypotensive response.2013 HUST-China iGEM use a synthetic way to combine bio-oscillator with propionate-producing enzy-
+					 was recently shown to produce an acute hypotensive response.2013 HUST-China iGEM use a synthetic way to combine bio-oscillator with propionate-producing enzyme
-					 me gene, trying to build a gut probiotic which can release pro-
+					 gene, trying to build a gut probiotic which can release propionate
-					 pionate periodically in accord with the rhythm of human BP. This could be a great substitute for chemical drugs by saving patients from drug dependence and the risk of sudden death at m-
+					 periodically in accord with the rhythm of human BP. This could be a great substitute for chemical drugs by saving patients from drug dependence and the risk of sudden death at
-					 orning BP peak time.
+					 morning BP peak time.
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 				<div class="clearfix"></div>
 			</div>
-			<div class="row" style="width:900px;margin:20px 0 30px 50px;">
+			<div class="row" style="width:900px;margin:20px 0 30px 50px;box-shadow:1px 1px 15px 5px;box-shadow:1px 1px 15px 5px rgba(0, 0, 0, 0.5)">
-				<div style="float:left;width:408px;height:400px;text-align:left;padding:0 20px;border:1px solid #fff;">
+				<div id="oscillator" style="float:left;width:408px;height:320px;text-align:left;padding:0 20px;border:1px solid #fff;">
 					<div>
 						<span style="font-size:36px;"><a href="https://2013.igem.org/Team:HUST-China/Project">Oscillator</a></span>
@@ Line 86: / Line 90: @@
 					</div>
 					<div>
-						The key part of the oscillator is araBAD/ lacZYA hybrid promoter. It is activated by the AraC protein in the pre-
-						sence of arabinose and repressed by the LacI protein in the absence of IPTG. Hence, activation of the promoters by the addition of Arabinose and IPTG to the medium r-
+						The key part of the oscillator is araBAD/ lacZYA hybrid p-
-						esults in transcription of each component of the circuit.
+						romoter. It is activated by the AraC protein in the prese-
-						Increased production of AraC in the presence of arabin-
+						nce of arabinose and repressed by the LacI protein in the absence of IPTG, constructing two feedback loops with opposite effects.And the differential activity of the two feedback loops can drive oscillatory behaviour.
-						ose results in a positive feedback loop that increases p-
-						romoter activity. However, the concurrent increase in production of LacI results in a linked negative feedback loop that decreases promoter activity. The differential activity of the two feedback loops can drive oscillatory behavior.
 					</div>
 				</div>
-				<div style="float:left;width:408px;height:400px;text-align:left;padding:0 20px;border:1px solid #fff;border-left:none;">
+				<div id="propionate" style="float:left;width:408px;height:320px;text-align:left;padding:0 20px;border:1px solid #fff;border-left:none;">
 					<div>
-						<span style="display:inline-block;font-size:32px;width:190px;margin-top:14px;"><a href="https://2013.igem.org/Team:HUST-China/Project">Propionate<br/><br/>reactor</a></span>
+						<span style="display:inline-block;font-size:32px;width:190px;margin-top:14px;"><a href="https://2013.igem.org/Team:HUST-China/Project">Propionate<br/><br/>Reactor</a></span>
 						<span><a href=""><a href="https://2013.igem.org/Team:HUST-China/Project"><img src="https://static.igem.org/mediawiki/2013/9/9a/HUST-hplc.jpg" style="width:190px;margin-left:10px;"></a></span>
 					</div>
 					<div>
-						A four gene operon in E.coli K12 genome which includes sbm ygfG ygfH ygfD, is significant in the metabolic pathway that converts succinate to propionate through Wood-Werkman reaction. We construct effective expression plasmid to increase the quantity of the four enzymes independently. By detecting the propionate amount by HPLC, we get to know which of the four is the rate-limiting enzyme. We choose it as the output of the oscillator.
+						A four-gene operon in E.coli K12 genome which includes sbm,ygfG,ygfH and ygfD, is significant in the metabolic pathway that converts succinate to propionate through Wood-Werkman reaction. We constructed effective expression plasmid to increase the quantity of the four enzymes independently. By measuring the propionate amount, we figured out which of the four is the most effective.
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-			<div class="row" style="width:900px;margin:20px 0 30px 50px;">
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-				<div style="float:left;width:408px;height:240px;text-align:left;padding:15px 20px 0;border:1px solid #fff;">
+				<div id="future-work" style="float:left;width:408px;height:240px;text-align:left;padding:15px 20px 0;border:1px solid #fff;">
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 						<span style="font-size:29px;"><a href="https://2013.igem.org/Team:HUST-China/Project">Future work</a></span>
@@ Line 112: / Line 114: @@
 					</div>
 					<div>
-						In the future, we will use the gene of rate-limiting enzyme of propionate producing reaction to replace mRFP in the oscillator as the output. And we hope to see periodical r-
+						In the near future, we will regulate the period of propionate utilizing the frequency divider with a ssrA-tag analog attached to the end of enzyme.Besides,we will replace mRFP with key gene in the synthetic pathway, hoping to see periodical release of propionate in accord with the rhythm of human BP.
-						elease of propionate.
-Later ,we may use mouse model to demonstrate our original design.
 					</div>
 				</div>
-				<div style="float:left;width:408px;height:255px;text-align:left;padding:0 20px;border:1px solid #fff;border-left:none;">
+				<div id="protocol" style="float:left;width:408px;height:255px;text-align:left;padding:0 20px;border:1px solid #fff;border-left:none;">
 					<div>
 						<span style="display:inline-block;font-size:37px;width:190px;margin-top:14px;"><a href="https://2013.igem.org/Team:HUST-China/Protocol">Protocol</a></span>
@@ Line 123: / Line 123: @@
 					</div>
 					<div>
-						Aside of traditional molecular biology method, we mainly use HPLC to detect propionate, flow cytometer(FCM) and confocal to detect fluorescence.
+						Our project is divided into three parts: the construction of the biological oscillator, the output evaluation of propionate by HPLC and the standardization of four genes.
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-			<div class="row" style="width:900px;margin:20px 0 30px 50px;">
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-				<div style="float:left;width:408px;height:320px;text-align:left;padding:15px 20px 0;border:1px solid #fff;">
+				<div id="modeling" style="float:left;width:408px;height:320px;text-align:left;padding:15px 20px 0;border:1px solid #fff;">
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 						<span style="font-size:35px;"><a href="https://2013.igem.org/Team:HUST-China/Modelling">Modeling</a></span>
@@ Line 135: / Line 135: @@
 					</div>
 					<div>
-						Detailed analysis of the oscillator makes us clearer about how it works. From the establishment of DDEs (delayed differential equations) to parameter sweep and sensitivity analysis, we know how each parameter contributes to the period. The MCOS (MultiCell Oscillation Simulation)shows us the exact feasibility of the oscillator both in vitro and in vivo (colon).
+						Detailed analysis of the oscillator makes us clearer about how it works. From the establishment of <acronym title="Delayed Differential Equations">DDEs</acronym> to parameter sweep and sensitivity analysis, we know how each parameter contributes to the period. The <acronym title="Multi Cells Oscillation Simulation">MCOS</acronym> shows us the feasibility of a group of oscillators that can be eventually applied in practice in vivo (colon).Lastly, we statistically analyzed our simulated data and wet-lab data.
 					</div>
 				</div>
-				<div style="float:left;width:408px;height:335px;text-align:left;padding:0 20px;border:1px solid #fff;border-left:none;">
+				<div id="human-practice" style="float:left;width:408px;height:335px;text-align:left;padding:0 20px;border:1px solid #fff;border-left:none;">
 					<div>
-						<span style="display:inline-block;font-size:35px;width:190px;margin-top:14px;"><a href="https://2013.igem.org/Team:HUST-China/HumanPractice">Human <br><br>practice</a></span>
+						<span style="display:inline-block;font-size:35px;width:190px;margin-top:14px;"><a href="https://2013.igem.org/Team:HUST-China/HumanPractice">Human <br><br>Practice</a></span>
 						<span><a href="https://2013.igem.org/Team:HUST-China/HumanPractice"><img src="https://static.igem.org/mediawiki/2013/1/11/HUST-humanpractice.jpg" style="width:190px;margin-left:10px;"></a></span>
 					</div>
 					<div>
-						Human practice:We have done a remarkable job in intro-
+						We have done a remarkable job in introducing high school students synthetic biology and
-						ducing high school students to synthetic biology and
+						iGEM jamborees as well as motivating them towards future participation in the iGEM. Besides, we collaborated with two other iGEM teams by sharing plasmids and characterizing their parts. Furthermore, we made  public speeches in our school about what we achieved,shared the felt in the iGEM competition.
-						iGEM jamborees as well as motivating them towards fu-
-						ture participation in the iGEM. Besides, we collaborated with two other iGEM teams by sharing DNA materials and characterizing their parts. Furthermore, we made a public speech in our school to report what we achieved, to sha-
-						re the feelings in the iGEM competition.And we also desi-
-						gn a crossword puzzle about iGEM for fun of all iGEMers.
 					</div>
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@@ Line 157: / Line 153: @@
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-                 <div class="modal-footer footer">
+                 <div class="modal-footer footer"><h2>Acknowledgement</h2><hr>
-			<div style="margin:0 auto 40px;width:530px;text-align:left">
+			<div style="margin:0 auto 40px;width:900px;text-align:left">
-					<strong>Acknowledgement</strong>
 					<p>
-						<span style="display:inline-block;float:left">
+                                                <span style="display:inline-block;float:left;margin-right:20px">
-							<a href="http://life.hust.edu.cn" target="_blank" title="Life Science and Technology of HUST"><img src="https://static.igem.org/mediawiki/2013/a/a5/HUST-life.png" width="100px" height="100px"/></a>
+							<a href="http://english.hust.edu.cn" target="_blank" title="Huazhong University of Science and Technology"><img src="https://static.igem.org/mediawiki/2013/0/00/HUST-hust.jpg" width="120px" height="100px"/></a>
 						</span>
-						<span style="display:inline-block;float:right">
+						<span style="display:inline-block;float:left;margin-right:20px">
+							<a href="http://life.hust.edu.cn" target="_blank" title="Life Science and Technology of HUST"><img src="https://static.igem.org/mediawiki/2013/a/a5/HUST-life.png" width="120px" height="100px"/></a>
+						</span>
+						<span style="display:inline-block;float:left;margin:0 20px 20px 0;">
 							<a href="http://qiming.hust.edu.cn" target="_blank" title="Qiming College of Huazhong University of Science and Technology"><img src="https://static.igem.org/mediawiki/2013/4/46/HUST-qiming.png" /></a>
 						</span>
+						<span style="display:inline-block;float:left;margin:0 20px 20px 0;">
+							<a href="javascript:void();" ><img src="https://static.igem.org/mediawiki/2013/6/6b/HUST-njsd.jpg" width="200px"/></a>
+						</span>
+						<span style="display:inline-block;float:left;margin-right:20px">
+							<a href="javascript:void();" ><img src="https://static.igem.org/mediawiki/2013/c/c7/HUST-bci.jpg" width="270px"/></a>
+						</span>
+						<span style="display:inline-block;float:left;margin-right:20px;">
+							<a href="javascript:void();" ><img src="https://static.igem.org/mediawiki/2013/5/51/HUST-ucsd.jpg" width="270px"/></a>
+						</span>
 					</p>
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-				<div class="clearfix"></div>
+				<div class="clearfix"></div><br>
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 					CopyRight &copy;<a href="https://2013.igem.org">2013.igem.org</a>&nbsp;All Rights Reserved!
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