Difference between revisions of "Team:Worldshaper-XSHS/design.html"

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                         <p class="v1" id="Ourteam" style="text-align:  center; ">
 
                         <p class="v1" id="Ourteam" style="text-align:  center; ">
                             Collaborations with other teams
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                             Design
 
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                        <p class="itemstyle">
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                            Based on our previous study, interviews and social investigation results, we planned to develop an efficient, affordable and convenient method to monitor the concentration of nicotine in the particular environment. If the idea of the experiment is proved feasible, the experimental results are likely to be directly applied into the environmental quality testing of relevant units. The first step in this plan is to construct plasmids that can express specifically in the presence of nicotine. After comprehending a great number of related researches and reading a plenty of related literatures, we finally found the Nox and NicA genes that bear on nicotine degradation, and predicted their promoters by literature【1】 and several websites. Then we named these promoters Pnox, Pnica1 and Pnica2 We hoped their promoters to be nicotine-induced and if fortunately the result is exactly as we expected, it would be possible for us to use the synthetic biological methods to construct E. coli strains as a component of detector.
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                         <p style="width: 800px;margin-left: 11px;margin-top: -23px;font-size: 22px;margin-bottom: 20px;">Collaboration with ASTWS-China team and HFLS_ZhejiangUnited team.</p>
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                         <p style="width: 800px;margin-left: 11px;margin-top: -23px;font-size: 22px;margin-bottom: 20px;">Design of the First Version</p>
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                        <p class="itemstyle">
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                            During the interlab experiment, we cooperated with ASTWS-China team and HFLS_ZhejiangUnited team, we provided the ASTWS-China team with E. coli DH5a and 100μl silicon beads that had better quality as well as gave HFLS_ZhejiangUnited team the transcriptional shaking table bacteria solution and some experimental equipment (chloramphenicol resistance, gun head, etc.). At the same time, we also borrowed the glycerin bacteria we needed from the ASTWS-China team.
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                     <div class="xshs-box3">
 
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                         <p class="itemstyle">
 
                         <p class="itemstyle">
                            During the experiment, we lent ASTWS-China team kit1 and kit2, which enables its students to extract the useful DNA.
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                          First of all, we designed the first version of the nicotine induced gene fragment, including: the predicted Nox promoter as a sensor, and the GFP gene as a reporter. In a single cell, different amounts of GFP will reflect nicotine concentrations in different nicotine solutions. We hope that nicotine concentration will be reflected in different fluorescence intensity eventually. Unfortunately, the part contains GFP gene could not be successfully transformed into E. coli, so our following experiment was shelved. Later we used RFP and BFP gene instead of GFP to reflect nicotine concentration. However, RFP gene doesn't contain RBS and terminator while BFP gene doesn't contain terminator, and may need later addition.
                        </p>
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                        <p class="itemstyle">
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                            During the Collaborations, our research members visited HFLS_ZhejiangUnited team’s experiments and watched its members’ studying notes.
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                         <p>
                             <img class="img-responsive" alt="" src="https://static.igem.org/mediawiki/2018/3/35/T--worldshaper-XSHS--c002.png" />
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                            <img class="img-responsive" alt="" src="https://static.igem.org/mediawiki/2018/a/ac/T--Worldshaper-XSHS--des005.png" />
                        <p class="itemstyle">
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                            Because our conversion process——A procedure that obtaining the desired plasmid from the distribution kits—— always failed,we communicated with ASTWS-China team, eliminating the possibilities derived from operate miss and finding the cause of the problem: Homemade E. coli receptive state.
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                         </p>
 
                         </p>
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                    <div class="xshs-box2">
 
 
                         <p>
 
                         <p>
                             <img class="img-responsive" alt="" src="https://static.igem.org/mediawiki/2018/b/bb/T--worldshaper-XSHS--c003.png" />
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                             <img class="img-responsive" alt="" src="https://static.igem.org/mediawiki/2018/3/3e/T--Worldshaper-XSHS--des006.png" />
 
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                     <div class="xshs-box3">
 
                     <div class="xshs-box3">
                        <p class="v1" id="OurSchool"style="text-align:  center; "> Collaborations with other teams</p>
 
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                            <p style="width: 800px;margin-left: 11px;margin-top: -23px;font-size: 22px;margin-bottom: 20px;">Collaboration with ZJU-China team.</p>
 
 
 
                         <p class="itemstyle">
 
                         <p class="itemstyle">
                             The aspect about experimental problem: Because of the invalid homemade E. coli receptive state, we did experiments in the ZJU-China team’s laboratories by utilizing its commercial E. coli receptive state. This attempt provided enough opportunities for us to discuss the difference between the experiments, which exerted great advantage to solve such problems in the nest phase.
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                             We found that the Nox promoter was not a nicotine specifically induced promoter, so we predicted and built two series of NicA promoters. At the same time, we were lucky to borrow the GFP gene transformed Escherichia coli strain, so that we designed two of the first version monitoring systems that using NicA promoter as the monitor, GFP gene as the reporter and containing the double terminator. The result shows that one sort of predicted NicA promoters could be specifically induced by nicotine.
 
                         </p>
 
                         </p>
 
                     </div>
 
                     </div>
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                     <div class="xshs-box2">
 
                     <div class="xshs-box2">
 
                         <p>
 
                         <p>
                             <img class="img-responsive" alt="" src="https://static.igem.org/mediawiki/2018/7/74/T--worldshaper-XSHS--c004.png" />
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                             <img class="img-responsive" alt="" src="https://static.igem.org/mediawiki/2018/5/5b/T--Worldshaper-XSHS--des001.png" />
 
                         </p>
 
                         </p>
                    </div>
 
                    <div class="xshs-box3">
 
                        <p class="itemstyle">
 
                            Before carrying out publicity activities, our team made comprehensive discussions with the ZJU-China team like establishing programs of activities, preparing materials, and decorating the venues.
 
                        </p>
 
                    </div>
 
                    <div class="xshs-box2">
 
 
                         <p>
 
                         <p>
                             <img class="img-responsive" alt="" src="https://static.igem.org/mediawiki/2018/e/e1/T--worldshaper-XSHS--c005.png" />
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                             <img class="img-responsive" alt="" src="https://static.igem.org/mediawiki/2018/7/7e/T--Worldshaper-XSHS--des002.png" />
 
                         </p>
 
                         </p>
 
                     </div>
 
                     </div>
 
  
 
                     <div class="xshs-box3">
 
                     <div class="xshs-box3">
                         <p class="v1" id="Others"style="text-align: center;">Collaborations with social organizations</p>
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                         <p class="itemstyle">
                        <div style="
+
                             With these three plasmids construction experiments, we hope to further improve the expression efficiency of GFP on the basis of the first generation detection system, so that we can more accurately detect nicotine concentration in the environment. With the help of Professor Zhu Xufen, we use T7 RNA polymerase gene and T7 strong promoter to improve the efficiency of GFP expression. Therefore, we hope that the gene fragments of the second generation detection system include: Pnica2 promoter, T7 RNA polymerase gene, T7 strong promoter and GFP coding gene. Pnica2 promoter does not directly control the expression of GFP, but directly controls the expression of T7RNA polymerase. The latter was consistent with the T 7 promoter and the expression efficiency was much higher than that of the other combinations. We expect that GFP, indirectly regulated by Pnica2 promoter, will have higher expression efficiency. Of course, later experimental results confirm that our expectation has been fulfilled.
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                        ">
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                        <div class="xshs-box3">
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                            <p style="width: 800px;margin-left: 11px;margin-top: -23px;font-size: 22px;margin-bottom: 20px;">Collaboration with Zhejiang Province Science Museum</p>
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                        </div>
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                        <p class="itemstyle" style="margin-left: 0px;width: 1100;">
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                          We cooperated with Science museum and did some simple experiments in the venue stadium, which is not only appeals visitors, but also facilitates to promote the IGEM project and knowledge about synthetic biology.
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                         </p>
 
                         </p>
                        <div class="xshs-box3">
 
                            <div style="
 
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                            <p style="width: 800px;margin-left: 11px;margin-top: -23px;font-size: 22px;margin-bottom: 20px;">Collaborations with Zhejiang Science and Technology Market</p>
 
                        </div>
 
                       
 
                        <p class="itemstyle" style="margin-left: 0px;width: 1100;">
 
                          During the science and technology week held by Zhejing Science administrators, we collaborated with organizers. In that exhibition, we conducted a questionnaire survey as well as advertised the JGEM project, thus, enriching the program’s content.
 
                        </p>
 
                     
 
 
                     </div>
 
                     </div>
                   
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                     <div class="xshs-box2">
 
                     <div class="xshs-box2">
 
                         <p>
 
                         <p>
                             <img class="img-responsive" alt="" src="https://static.igem.org/mediawiki/2018/a/a6/T--worldshaper-XSHS--c006.png" />
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                             <img class="img-responsive" alt="" src="https://static.igem.org/mediawiki/2018/4/44/T--Worldshaper-XSHS--des003.png" />
 
                         </p>
 
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Revision as of 12:43, 17 October 2018

Design

Design


Based on our previous study, interviews and social investigation results, we planned to develop an efficient, affordable and convenient method to monitor the concentration of nicotine in the particular environment. If the idea of the experiment is proved feasible, the experimental results are likely to be directly applied into the environmental quality testing of relevant units. The first step in this plan is to construct plasmids that can express specifically in the presence of nicotine. After comprehending a great number of related researches and reading a plenty of related literatures, we finally found the Nox and NicA genes that bear on nicotine degradation, and predicted their promoters by literature【1】 and several websites. Then we named these promoters Pnox, Pnica1 and Pnica2 We hoped their promoters to be nicotine-induced and if fortunately the result is exactly as we expected, it would be possible for us to use the synthetic biological methods to construct E. coli strains as a component of detector.

Design of the First Version

First of all, we designed the first version of the nicotine induced gene fragment, including: the predicted Nox promoter as a sensor, and the GFP gene as a reporter. In a single cell, different amounts of GFP will reflect nicotine concentrations in different nicotine solutions. We hope that nicotine concentration will be reflected in different fluorescence intensity eventually. Unfortunately, the part contains GFP gene could not be successfully transformed into E. coli, so our following experiment was shelved. Later we used RFP and BFP gene instead of GFP to reflect nicotine concentration. However, RFP gene doesn't contain RBS and terminator while BFP gene doesn't contain terminator, and may need later addition.

We found that the Nox promoter was not a nicotine specifically induced promoter, so we predicted and built two series of NicA promoters. At the same time, we were lucky to borrow the GFP gene transformed Escherichia coli strain, so that we designed two of the first version monitoring systems that using NicA promoter as the monitor, GFP gene as the reporter and containing the double terminator. The result shows that one sort of predicted NicA promoters could be specifically induced by nicotine.

With these three plasmids construction experiments, we hope to further improve the expression efficiency of GFP on the basis of the first generation detection system, so that we can more accurately detect nicotine concentration in the environment. With the help of Professor Zhu Xufen, we use T7 RNA polymerase gene and T7 strong promoter to improve the efficiency of GFP expression. Therefore, we hope that the gene fragments of the second generation detection system include: Pnica2 promoter, T7 RNA polymerase gene, T7 strong promoter and GFP coding gene. Pnica2 promoter does not directly control the expression of GFP, but directly controls the expression of T7RNA polymerase. The latter was consistent with the T 7 promoter and the expression efficiency was much higher than that of the other combinations. We expect that GFP, indirectly regulated by Pnica2 promoter, will have higher expression efficiency. Of course, later experimental results confirm that our expectation has been fulfilled.

  • Worldshaper-XSHS, Xiaoshan High School

    Adress: No.538,Gongxiu Road,Xiaoshan District,Hangzhou,Zhejiang Province,China