Team:Shenzhen SFLS/Project

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<h1 class="hide">Project</h1>
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You are provided with this team page template with which to start the iGEM season. You may choose to personalize it to fit your team but keep the same "look." Or you may choose to take your team wiki to a different level and design your own wiki.  You can find some examples <a href="https://2009.igem.org/Help:Template/Examples">HERE</a>.
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            <div class="project">
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You <strong>MUST</strong> have the following information on your wiki:
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<li>a team description</li>  
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<p>This is our project, EcoPi. <br />
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<li>project description</li>
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<img src="https://static.igem.org/mediawiki/2013hs/4/41/Project_img_1.jpg" width="800" height="262" alt="" /> 
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<li>safety information (did your team take a safety training course? were you supervised in the lab?)</li>
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</p>
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<li>team attribution (who did what part of your project?)</li>
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<p>In this page, you can see how it was designed and made.</p>
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</ul>
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<p>Eutrophication can be a very severe problem to the environment. In an effort to solve this problem, we want to build a genetic system which can test and remove the phosphate from water. In our idea, we want to make a product which can automatically detects phosphate and therefore remove it. And it must be a totally environmentally friendly one. And we began our designing.
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Frist we designed a genetic circuit which can test how the work has been done. We wanted to find a promoter which can only be induced by Pi starvation. Then there is a suppressor protein and an <span>rfp</span>. A double terminator is at the end of this device. This device can only work when the concentration of Pi is low. And the water will show red fluorescence. The red can also be a signal of ‘STOP’, which means the work has already finished.<br />
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<li>lab notebook</li>
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<img src="https://static.igem.org/mediawiki/2013hs/e/e2/Project_img_3.jpg" width="800" height="244" alt="" />
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<li>sponsor information</li>
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Next we designed a second device for removing the phosphate. The promoter of this device is repressed by the protein in Device one, which means when the concentration of Pi is low, Device two will be limited and stop working. Then we wanted to find a CDS which can remove phosphate from water. And a <span>gfp</span>+LVA(degradation tag) with a terminator at the end. When this device is working, The CDS will remove the phosphate from water and meanwhile the gfp+LVA will show green fluorescence, just like the green traffic signal. This device is mainly used for removing phosphate.
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<li>other information</li>
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</p>
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</ul>  
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<p>This photo can show you how the two device are linked
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<img src="https://static.igem.org/mediawiki/2013hs/8/87/Project_img_4.jpg" width="800" height="525" alt="" />
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When we put our product into the water which has a very high concentration of phosphate, Device one will be limited and Device two will be activated. Then the water will show green fluorescence which means our product is removing Pi.
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<img src="https://static.igem.org/mediawiki/2013hs/b/bf/Project_img_5.jpg" width="800" height="458" alt="" />
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After a period of time, due to Device two’s work, the concentration of Pi decreases, then Device one will be activated because of Pi starvation. The suppressor protein will repress Device two. Device one will work and show red fluorescence to tell us that the work has been finished.
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</p>
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<p>That’s the idea of our project, we then started searching for files that support our idea. First we searched for a CDS which can digest phosphate. Then after weeks of searching we found Polyphosphate Kinase, also known as PPK. It is a kind of enzyme widely distributed in ecosystem, more importantly, it can be found in E-coli. Here this picture below can show you how it works.
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<img src="https://static.igem.org/mediawiki/2013hs/c/cd/Project_img_6.jpg" width="800" height="298" alt="" />
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</p>
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<p>We found its genetic sequence and we used PCR to made it a standard biobrick. Then we started to find the promoters of our product. Finally we found the Pi Starvation Promoter(BBa_K737023), it’s not on the kits so we again designed its primer and used PCR.</p>
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<p>For the protein we used TetR protein(BBa_C0040)</p>
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<p>Now it’s time for our final genetic circuit. Take a look!
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<img src="https://static.igem.org/mediawiki/2013hs/7/72/Project_img_7.jpg" width="800" height="262" alt="" />
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</p>
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<p>Result</p>
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</p>
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<p>After we finished our project, we started to test it. We test two part of our project. The first one is our Device One. We tested its response to Pi starvation. Here is the results diagram.</p>
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</p>
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<p>In this diagram we can find that Device one can be induced by phosphate starvation. When the concentration of dipotassium phosphate is 0.03μM we can see that our sensor reached a peak because our promoter can be activated by Pi starvation.</p>
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</p>
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<p>Next is our whole Device. We tested when the green fluorescence would vanish and red fluorescence occur. Here is another diagram.</p>
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</p>
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<p>This diagram shows that after several hours the amount of green fluorescence decreases, which means the work has been done and red fluorescence will occur.</p>
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</p>
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<p>This result clearly pointed out that our product can work smoothly. The Pi starvation promoter is working well too. Our experiments have reached a success at last.</p
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{|align="justify"
 
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|You can write a background of your team here.  Give us a background of your team, the members, etc.  Or tell us more about something of your choosing.
 
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|[[Image:Deerfield_MA_logo.png|200px|right|frame]]
 
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''Tell us more about your project.  Give us background.  Use this as the abstract of your project.  Be descriptive but concise (1-2 paragraphs)''
 
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|[[Image:Shenzhen_SFLS.png|right|frame|Your team picture]]
 
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|align="center"|[[Team:Shenzhen_SFLS | Team Shenzhen_SFLS]]
 
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{| style="color:#1b2c8a;background-color:#0c6;" cellpadding="3" cellspacing="1" border="1" bordercolor="#fff" width="62%" align="center"
 
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!align="center"|[https://igem.org/Team.cgi?year=2013&division=high_school&team_name=Shenzhen_SFLS Official Team Profile]
 
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===Team===
 
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Peilin Li: the instructor of SFLS students.
 
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Kang Kang: the advisor of SFLS students. In BGI, K2 is veeeeery famous for lurking on BBS and sarcasm! As a former champion in a national robot contest, a hacker, a web engineer, a designer, a magazine editor, a citizen reporter, a filmmaker and a drama actor, he has always too many wonderings, including how life works on its core database - genome. Cracking and hacking the code of life, making actors in cell perform on his scripts, has become the primary task of this young scientist.
 
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Hekang Jia: the captain of SFLS students, set his sights on get the only grand prize. The boy who set up the team by himself，has strong desire to make everthing best. He was often very serious in team's seminar and made others under pressure. He is the also the leader of team's students working on theoretical derivation.
 
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Biwei Zheng: the vice captain of SFLS students.
 
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Ruping Liu: Conscientious and careful to anything. She’s interested in biology, not only because of its diversity but also its mystery. Her dream is to do something which is useful to the world in the future. The girl is working on reading paper and do some experiments in SFLS iGEM team.
 
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Kang Li: Eric Lee is a senior one student in SFLS and is responsible for the team’s theoretical group. He has a strong desire in biology and he is very good at communicating with people. So he is also responsible for communicating with the committee. He is a good speaker too. Eric is looking forward to helping the team more.
 
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Ziying Tan: Tina is studying in SFLS Class 7 Grade 10. Aside from spending time before the computer or in the lab, she enjoys rubber sculpture and reading.She is now one of the leaders of vacation club. She couldn`t think of a better way to enjoy her summer in school than joining Shenzhen SFLS IGEM team!
 
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===Project===
 
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Student at SFLS want to use an enzyme known as “polyphosphate kinase”（PPK）to governance the eutrophication of water bodies. We know that Inorganic phosphate (Pi) is recognized as one of the major nutrients contributing to the outbreak of red tide.We have found that PPK is known to digest Inorganic phosphate (Pi). There is a small amount of it in e.coli and we want to copy the gene of PPK in the e.coli to make it more efficient,so the role of PPK will be more obvious Then we will use the GFP to express the result of the digest of PPK.We want to make two device in a gene circut to deal with this problem.
 
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Device 1: We use a limitable promoter which would be limited by high concentrations of Pi, then the GFP in this device won't light and a protein we add in the device won't breed .
 
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Device 2: If the protein didn't breed, the promoter in Device 2 will be induced and the PPK gene will work, it will digest Pi. When the concentration of phosphate become low, devive 2 will stop to digest Pi and the promoter in device 1 will be induced, then the GFP in device 1 will light. That means the work has finished.
 
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( We will put on our picture of gene circuts in few days)
 
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===Notebook===
 
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Show us how you spent your days.
 
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===Results/Conclusions===
 
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What did you achieve over the course of your semester?
 
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===Safety===
 
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What safety precautions did your team take? Did you take a safety training course? Were you supervised at all times in the lab?
 
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===Attributions===
 
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Who worked on what?
 
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===Human Practices===
 
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What impact does/will your project have on the public? 
 
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===Fun!===
 
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What was your favorite team snack?? Have a picture of your team mascot?
 
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<forum_subtle />
 

Latest revision as of 02:35, 22 June 2013

Project

This is our project, EcoPi.

In this page, you can see how it was designed and made.

Eutrophication can be a very severe problem to the environment. In an effort to solve this problem, we want to build a genetic system which can test and remove the phosphate from water. In our idea, we want to make a product which can automatically detects phosphate and therefore remove it. And it must be a totally environmentally friendly one. And we began our designing. Frist we designed a genetic circuit which can test how the work has been done. We wanted to find a promoter which can only be induced by Pi starvation. Then there is a suppressor protein and an rfp. A double terminator is at the end of this device. This device can only work when the concentration of Pi is low. And the water will show red fluorescence. The red can also be a signal of ‘STOP’, which means the work has already finished.
Next we designed a second device for removing the phosphate. The promoter of this device is repressed by the protein in Device one, which means when the concentration of Pi is low, Device two will be limited and stop working. Then we wanted to find a CDS which can remove phosphate from water. And a gfp+LVA(degradation tag) with a terminator at the end. When this device is working, The CDS will remove the phosphate from water and meanwhile the gfp+LVA will show green fluorescence, just like the green traffic signal. This device is mainly used for removing phosphate.

This photo can show you how the two device are linked When we put our product into the water which has a very high concentration of phosphate, Device one will be limited and Device two will be activated. Then the water will show green fluorescence which means our product is removing Pi.

After a period of time, due to Device two’s work, the concentration of Pi decreases, then Device one will be activated because of Pi starvation. The suppressor protein will repress Device two. Device one will work and show red fluorescence to tell us that the work has been finished.

That’s the idea of our project, we then started searching for files that support our idea. First we searched for a CDS which can digest phosphate. Then after weeks of searching we found Polyphosphate Kinase, also known as PPK. It is a kind of enzyme widely distributed in ecosystem, more importantly, it can be found in E-coli. Here this picture below can show you how it works.

We found its genetic sequence and we used PCR to made it a standard biobrick. Then we started to find the promoters of our product. Finally we found the Pi Starvation Promoter(BBa_K737023), it’s not on the kits so we again designed its primer and used PCR.

For the protein we used TetR protein(BBa_C0040)

Now it’s time for our final genetic circuit. Take a look!

Result

After we finished our project, we started to test it. We test two part of our project. The first one is our Device One. We tested its response to Pi starvation. Here is the results diagram.

In this diagram we can find that Device one can be induced by phosphate starvation. When the concentration of dipotassium phosphate is 0.03μM we can see that our sensor reached a peak because our promoter can be activated by Pi starvation.

Next is our whole Device. We tested when the green fluorescence would vanish and red fluorescence occur. Here is another diagram.

This diagram shows that after several hours the amount of green fluorescence decreases, which means the work has been done and red fluorescence will occur.

This result clearly pointed out that our product can work smoothly. The Pi starvation promoter is working well too. Our experiments have reached a success at last.

Retrieved from "http://2013hs.igem.org/Team:Shenzhen_SFLS/Project"