Difference between revisions of "Team:Fudan-TSI/Parts"

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                <li style="padding: 0"><div class="sidenavBanner">
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                    <div class="background">
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                    </div>
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                    <p style="width: 100%;text-align: center;font-size: 24px"><span class="white-text">Parts</span></p>
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                </div></li>
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                <li>
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                    <ul class="collapsible expandable">
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                        <li>Team: Fudan-TSI</li>
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<li><div class="collapsible-header">Project</div>
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    <div class="collapsible-body"><ul>
 +
        <li><a href="/Team:Fudan-TSI/Description">Background</a></li>
 +
        <li><a href="/Team:Fudan-TSI/Design">Design</a></li>
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        <li><a href="/Team:Fudan-TSI/Applied_Design">Applied Design</a></li>
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        <li><a href="/Team:Fudan-TSI/Experiments">Experiments</a></li>
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        <li><a href="/Team:Fudan-TSI/Judging">Judging</a></li>
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    </ul></div>
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</li>
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<li><div class="collapsible-header">Results</div>
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    <div class="collapsible-body"><ul>
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        <li><a href="/Team:Fudan-TSI/Results#ReverseTranscription">Reverse Transcription</a></li>
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        <li><a href="/Team:Fudan-TSI/Results#Recombination">Recombination</a></li>
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        <li><a href="/Team:Fudan-TSI/Demonstrate">Demonstration</a></li>
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        <li><a href="/Team:Fudan-TSI/Measurement">Measurement</a></li>
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        <li><a href="/Team:Fudan-TSI/Notebook">Notebook</a></li>
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    </ul></div>
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</li>
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<li><div class="collapsible-header">Model</div>
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    <div class="collapsible-body"><ul>
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        <li><a href="/Team:Fudan-TSI/Model">Modeling</a></li>
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        <li><a href="/Team:Fudan-TSI/Software">Software</a></li>
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        <li><a href="/Team:Fudan-TSI/Hardware">Hardware</a></li>
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    </ul></div>
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</li>
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<li><div class="collapsible-header">Parts</div>
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    <div class="collapsible-body"><ul>
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        <li><a href="/Team:Fudan-TSI/Basic_Part">Basic parts</a></li>
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        <li><a href="/Team:Fudan-TSI/Composite_Part">Composite parts</a></li>
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        <li><a href="/Team:Fudan-TSI/Improve">Improved parts</a></li>
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        <li><a href="/Team:Fudan-TSI/Part_Collection">Part collection</a></li>
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    </ul></div>
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</li>
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<li><div class="collapsible-header">Outreach</div>
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    <div class="collapsible-body"><ul>
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        <li><a href="/Team:Fudan-TSI/Public_Engagement">Education &amp; Public engagement</a></li>
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        <li><a href="/Team:Fudan-TSI/Integrated_Human_Practice">Integrated human practice</a></li>
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        <li><a href="/Team:Fudan-TSI/Collaborations">Collaborations</a></li>
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        <li><a href="/Team:Fudan-TSI/Safety">Safety</a></li>
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    </ul></div>
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</li>
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<li><div class="collapsible-header">Team</div>
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    <div class="collapsible-body"><ul>
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        <li><a href="/Team:Fudan-TSI/Team">Members</a></li>
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        <li><a href="/Team:Fudan-TSI/Attributions">Attributions</a></li>
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        <li><a href="https://2018.igem.org/Team:Fudan/Heritage" target=_blank>Heritage</a></li>
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        <li><a href="/Team:Fudan-TSI">&copy; 2019</a></li>
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    </ul></div>
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                         </p>
 
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                          <p>PCR and subcloning were performed using standard methods. Detailed primer sequences are <a href="/Team:Fudan-TSI/Primers">provided</a>. All constructs were verified by Sanger sequencing.
 
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                          </p>
 
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                          <p>Cells were cultured in DMEM supplemented with 10% FBS (HyClone), 100 U/ml penicillin, 100 μg/ml streptomycin and 1x GlutaMax (Gibco). Transient transfections were performed using Lipofectamine 2000 (Invitrogen) and Opti-MEM (Gibco). Viral packaging, infection and fluorescence-activated cell sorting were performed using standard methods.
 
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                          </p>
 
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                          <p>Images, unless otherwise indicated, were captured using an inverted epifluorescence microscope (IX-81, Olympus) and a sCMOS camera (pixel size = 0.3222 &mu;m; Zyla 5.5, Andor; 20x objective N.A.  0.75) and were controlled by <a href="http://www.micro-manager.org/" target=_blank>Micro-Manager software</a>.
 
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                          </p>
 
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                          <p>All statistical analysis was performed using Prism (Graphpad) and <a href="http://rsbweb.nih.gov/ij/developer/macro/macros.html" target=_blank>ImageJ</a>. All experiments were independently performed in triplicates; unless otherwise indicated. Images were combined and annotated in Powerpoint for presentation. Representative images are shown.
 
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                                  <th>Our protocols as PDF files</th><th>&nbsp;</th>
 
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                                  <th>Download</th>
 
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                              </tr>
 
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                              <tr>
 
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                                  <td>Be a Good Lab Member</td><td><i>GoodLabPractices.pdf</i></td>
 
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                                  <td><a href="https://2018.igem.org/File:T--Fudan--GoodLabPractices.pdf" target="_blank"><i class="fa fa-download"></i></a></td>
 
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                              </tr>
 
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                              <tr>
 
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                                  <td>Molecular Cloning</td><td><i>MolecularCloning.pdf</i></td>
 
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                                  <td><a href="https://2018.igem.org/File:T--Fudan--MolecularCloning.pdf" target="_blank"><i class="fa fa-download"></i></a></td>
 
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                              </tr>
 
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                                  <td>Tissue Culture</td><td><i>CellCulture.pdf</i></td>
 
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                                  <td><a href="https://2018.igem.org/File:T--Fudan--CellCulture.pdf" target="_blank"><i class="fa fa-download"></i></a></td>
 
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                              </tr>
 
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                                  <td>Make a Stable Cell Line</td><td><i>MakeStableCellLine.pdf</i></td>
 
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                                  <td><a href="https://2018.igem.org/File:T--Fudan--MakeStableCellLine.pdf" target="_blank"><i class="fa fa-download"></i></a></td>
 
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                              </tr>
 
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                              <tr>
 
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                                  <td>Cell Sorting</td><td><i>FACS.pdf</i></td>
 
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                                  <td><a href="https://2018.igem.org/File:T--Fudan--FACS.pdf" target="_blank"><i class="fa fa-download"></i></a></td>
 
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                              </tr>
 
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                                  <td>Cell Staining</td><td><i>FixStain.pdf</i></td>
 
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                                  <td><a href="https://2018.igem.org/File:T--Fudan--FixStain.pdf" target="_blank"><i class="fa fa-download"></i></a></td>
 
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                                  <td>Time-lapse Live-cell Imaging</td><td><i>TimeLapseImaging.pdf</i></td>
 
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                                  <td><a href="https://2018.igem.org/File:T--Fudan--TimeLapseImaging.pdf" target="_blank"><i class="fa fa-download"></i></a></td>
 
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                              </tr>
 
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                          </table>
 
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                      </div>
 
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                      <p style="color:grey">
 
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                          For practical reasons, all full-length protocols are in Chinese.
 
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                      </p>
 
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                  </div>
 
  
 
                 </main>
 
                 </main>
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                 <div class="container">
 
                 <div class="container">
 
                     <h2 style="margin: 0;padding: 10px 0;">Project Summary</h2>
 
                     <h2 style="margin: 0;padding: 10px 0;">Project Summary</h2>
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                     <p style="margin: 0">abstract abstract abstract abstract abstract abstract abstract abstract abstract abstract abstract abstract abstract abstract abstract abstract due sep 5 due sep 5 due sep 5 due sep 5 due sep 5 due sep 5 due sep 5 due sep 5 due sep 5 due sep 5 due sep 5 due sep 5
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                     <p style="margin: 0">Mutation library generation is critical for biological and medical research, but current methods cannot mutate a specific sequence continuously without manual intervention. Here we present a toolbox for <i>in vivo</i> continuous mutation library construction. First, the target DNA is transcribed into RNA. Next, our reverse transcriptase reverts RNA into cDNA, during which the target is randomly mutated by enhanced error-prone reverse transcription. Finally, the mutated version replaces the original sequence through recombination. These steps will be carried out iteratively, generating a random mutation library of the target with high efficiency as mutations accumulate along with bacterial growth. Our toolbox is orthogonal and provides a wide range of applications among various species. R-Evolution could mutate coding sequences and regulatory sequences, which enables the <i>in vivo</i> evolution of individual proteins or multiple targets at a time, promotes high-throughput research, and serves as a foundational advance to synthetic biology.
 
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                         <div id="sponsor" class="col m3 s12 row">
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                             <a href="https://2019.igem.org/Team:Fudan"><img alt="2018 Team:Fudan logo white" class="col s3 m6 l3" style="position:relative; padding: 0.5em 0.3rem; margin:-0.15rem 0; left: -0.45rem;" src="https://static.igem.org/mediawiki/2019/7/7f/T--Fudan-TSI--Logo0-crop-grey.png">
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                             <a href="https://2019.igem.org/Team:Fudan-TSI"><img alt="2018 Team:Fudan logo white" class="col s3 m6 l3" style="position:relative; padding: 0.5em 0.3rem; margin:-0.15rem 0; left: -0.45rem;" src="https://static.igem.org/mediawiki/2019/7/7f/T--Fudan-TSI--Logo0-crop-grey.png">
 
                             </a><a href="http://www.fudan.edu.cn/en/" target="_blank"><img class="col s3 m6 l3" alt="Fudan University" src="https://static.igem.org/mediawiki/2018/f/f7/T--Fudan--schoolLogo.png">
 
                             </a><a href="http://www.fudan.edu.cn/en/" target="_blank"><img class="col s3 m6 l3" alt="Fudan University" src="https://static.igem.org/mediawiki/2018/f/f7/T--Fudan--schoolLogo.png">
 
                         </a><a href="http://life.fudan.edu.cn/" target="_blank"><img class="col s3 m6 l3" style="margin-bottom: 4%;/* 该图比其他小一点，排版需要 */" alt="School of Life Sciences, Fudan University" src="https://static.igem.org/mediawiki/2018/1/1d/T--Fudan--schoolOfLifeSciencesIcon.png">
 
                         </a><a href="http://life.fudan.edu.cn/" target="_blank"><img class="col s3 m6 l3" style="margin-bottom: 4%;/* 该图比其他小一点，排版需要 */" alt="School of Life Sciences, Fudan University" src="https://static.igem.org/mediawiki/2018/1/1d/T--Fudan--schoolOfLifeSciencesIcon.png">
 
                         </a><a href="http://www.yfc.cn/en/" target="_blank"><img class="col s3 m6 l3" style="padding: 0.15rem 0.9rem;" alt="Yunfeng Capital" src="https://static.igem.org/mediawiki/2018/e/e2/T--Fudan--yunfengLogo.png">
 
                         </a><a href="http://www.yfc.cn/en/" target="_blank"><img class="col s3 m6 l3" style="padding: 0.15rem 0.9rem;" alt="Yunfeng Capital" src="https://static.igem.org/mediawiki/2018/e/e2/T--Fudan--yunfengLogo.png">
 
                         </a>
 
                         </a>
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                             <h3 class="col s12" style="text-align: left; color: rgba(255, 255, 255, 0.8); font-size: 18px">Repeated Evolution in vivo</h3>
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                             <h3 class="col s12" style="text-align: left; color: rgba(255, 255, 255, 0.8); font-size:12px">R-Evolution: an <i>in vivo</i> sequence-specific toolbox for continuous mutagenesis</h3>
 
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                         <div id="usefulLinks" class="col m9 s12 row">
 
                         <div id="usefulLinks" class="col m9 s12 row">
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                                   <span><a href="/Team:Fudan-TSI/Parts">Parts</a></span>
 
                                   <span><a href="/Team:Fudan-TSI/Parts">Parts</a></span>
 
                                     <ul>
 
                                     <ul>

Revision as of 05:06, 15 September 2019

2019 Team:Fudan-TSI Parts

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2018 team Fudan title attributions

text from greatbay

Parts Overview with 19 composite parts, and 36 basic parts. Don't think we win at quantity but we actually provided a series of data with high-quality. In the production of geraniol in E.coli, our favourite basic part is BBa_K2753003, pSB1C3-obGES cds. We have also created and synthesized competent pathways in yeast to convert geraniol to neptalactol. Besides, based on previous part like BBa_J23119, we generated a stabilised version by adding an up-element, a transcription-activator-like effector(TALE) binding site, and a genetic circuit encoding for TALE2. In addition, We created a novel, well-characterized, and well-documented TALE stabilized promoter (TALEsp) collection as well as two parts enabling future teams to create more stabilized promoters. This promoter library provides a reliable tool which allows teams to construct predictable and robust synthetic systems. Check the GreatBay_China's parts family! Part Table Name Type Description Designer Length W BBa_K2753002 Coding pSB1C3-GPPS cds WEI KUANGYI 894 W BBa_K2753003 Coding pSB1C3-obGES cds WEI KUANGYI 1704 W BBa_K2753018 Composite TALE2 sp1 WEI KUANGYI 3166 W BBa_K2753023 Composite TALE2 sp6 WEI KUANGYI 3164 W BBa_K2753030 Composite TALE1 sp1 WEI KUANGYI 3166 W BBa_K2753010 Composite pSB1C3-TDH3 promoter-G8H-tENO2 WEI KUANGYI 2374 W BBa_K2753011 Composite pSB1C3-TEF1 promoter-GOR-tENO1 WEI KUANGYI 1787 W BBa_K2753012 Composite pSB1C3-pRPL18P promoter-CrISY-tTDH1 WEI KUANGYI 2127 W BBa_K2753013 Composite pSB1C3-pTALE1-CP WEI KUANGYI 3956 W BBa_K2753014 Composite pSB1C3-pTALE2-CP WEI KUANGYI 3956 W BBa_K2753015 Composite pSB1C3-RiboJ-rbs-GPPS-rbs-ObGES WEI KUANGYI 2781 W BBa_K2753016 Composite pSB1C3-RiboJ-rbs-GPPS-GES-double terminator WEI KUANGYI 4028 W BBa_K2753019 Composite TALE2 sp2 WEI KUANGYI 3166 W BBa_K2753020 Composite TALE2 sp3 WEI KUANGYI 3166 W BBa_K2753021 Composite TALE2 sp4 WEI KUANGYI 3160 W BBa_K2753022 Composite TALE2 sp5 WEI KUANGYI 3160 W BBa_K2753024 Composite TALE1 sp2 WEI KUANGYI 3160 W BBa_K2753025 Composite TALE1 sp3 WEI KUANGYI 3166 W BBa_K2753026 Composite TALE1 sp4 WEI KUANGYI 3160 W BBa_K2753027 Composite TALE1 sp5 WEI KUANGYI 3160 W BBa_K2753029 Composite TALE1 sp6 WEI KUANGYI 3160 U W BBa_K2753032 Terminator L3S2P21 WEI KUANGYI 60 U W BBa_K2753033 Regulatory PT7A1w2 WEI KUANGYI 69 U W BBa_K2753034 Regulatory PT7A1w3 WEI KUANGYI 69 U W BBa_K2753035 Tag SarJ WEI KUANGYI 79 U W BBa_K2753036 RBS RBSsp1 WEI KUANGYI 20 U W BBa_K2753037 RBS RBSsp2 WEI KUANGYI 20 U W BBa_K2753038 Coding TALE1 WEI KUANGYI 2640 U W BBa_K2753039 Coding TALEsp2 WEI KUANGYI 2640 U W BBa_K2753043 Regulatory promoter sp1 WEI KUANGYI 79 U W BBa_K2753044 Regulatory TDH3 promoter WEI KUANGYI 667 U W BBa_K2753045 Regulatory TEF1 promoter WEI KUANGYI 408 U W BBa_K2753046 Regulatory pRPL18P promoter WEI KUANGYI 709 U W BBa_K2753047 Coding G8H WEI KUANGYI 1482 U W BBa_K2753048 Coding GOR WEI KUANGYI 1137 U W BBa_K2753049 Coding CrISY WEI KUANGYI 1167 U W BBa_K2753050 Terminator tENO2 WEI KUANGYI 225 U W BBa_K2753051 Terminator tENO1 WEI KUANGYI 242 U W BBa_K2753052 Terminator tTDH1 WEI KUANGYI 251 U W BBa_K2753053 RBS RBStrGPPS WEI KUANGYI 56 U W BBa_K2753054 RBS Charles RBS WEI KUANGYI 14 U W BBa_K2753055 Regulatory UPJ23119 WEI KUANGYI 55 U W BBa_K2753056 Regulatory promoter sp2 WEI KUANGYI 79 U W BBa_K2753057 Regulatory PUPAsp2 WEI KUANGYI 79 U W BBa_K2753058 Regulatory PUPAAsp2 WEI KUANGYI 79 U W BBa_K2753059 Regulatory PDNAsp2 WEI KUANGYI 73 U W BBa_K2753060 Regulatory PDNAsp2w WEI KUANGYI 73 U W BBa_K2753061 Regulatory SP119 WEI KUANGYI 77 U W BBa_K2753062 Regulatory Psp1 mutated WEI KUANGYI 73 U W BBa_K2753063 Regulatory PUPsp1 mutated WEI KUANGYI 79 U W BBa_K2753064 Regulatory PDNsp1 WEI KUANGYI 73 U W BBa_K2753065 Regulatory PDNsp1w WEI KUANGYI 73 U W BBa_K2753066 Regulatory Psp1 WEI KUANGYI 73 U W BBa_K2753067 Regulatory Psp1w WEI KUANGYI 73 U W BBa_K2753070 RBS RBSobGES WEI KUANGYI 24 Go to Basic Parts Go to Composite Parts Go to Part Collection Go to Improve

title

Fangfei Ye is responsible for all art design, which includes our team logo, team flag, team uniform (Dr. Cai gave comments), team name card, brochures, our posters, as well as materials related to our human practice events.

project summary

Project Summary

Mutation library generation is critical for biological and medical research, but current methods cannot mutate a specific sequence continuously without manual intervention. Here we present a toolbox for in vivo continuous mutation library construction. First, the target DNA is transcribed into RNA. Next, our reverse transcriptase reverts RNA into cDNA, during which the target is randomly mutated by enhanced error-prone reverse transcription. Finally, the mutated version replaces the original sequence through recombination. These steps will be carried out iteratively, generating a random mutation library of the target with high efficiency as mutations accumulate along with bacterial growth. Our toolbox is orthogonal and provides a wide range of applications among various species. R-Evolution could mutate coding sequences and regulatory sequences, which enables the in vivo evolution of individual proteins or multiple targets at a time, promotes high-throughput research, and serves as a foundational advance to synthetic biology.