|
|
Line 1: |
Line 1: |
| {{IGEM_TopBar}} | | {{IGEM_TopBar}} |
| {{Fudan}} | | {{Fudan}} |
− | <html> | + | <html> |
| <link rel="stylesheet" href="/wiki/index.php?title=Template:Fudan/materialize.min.css&action=raw&ctype=text/css"> | | <link rel="stylesheet" href="/wiki/index.php?title=Template:Fudan/materialize.min.css&action=raw&ctype=text/css"> |
| <style> | | <style> |
Line 221: |
Line 221: |
| html代码不在乎多个空格,仅显示一个 | | html代码不在乎多个空格,仅显示一个 |
| --> | | --> |
− | <h1>Our part collection offers a complete array of parts for further researchers to develop other probiotic secreted proteins/peptides.</h1> | + | <h1 tabIndex="1">Our part collection offers a complete array of parts for future researchers to develop other probiotic secreted proteins/peptides.</h1> |
| <div class="column full_size"> | | <div class="column full_size"> |
| + | <p>Updated on 2020-11-22: we were nominated for the Best Part Collection prize.</p> |
| <p>In the collection, we have secreted mcbABCD to help its colonization in the gut, and secreted CaAP to facilitate calcium uptake by gut cells. We test all parts used in these two circles: 5 signal peptide for secretion, 14 promoters for different strength expression (previously publish and available in <a href="https://addgene.org" target="_blank">addgene.org</a> but not in <a href="http://parts.igem.org/Main_Page" target="_blank">the Registry</a>), IPTG inducible promoter driven various protein coding sequences (confirmed individually before assembled together), and quorum-sensing components. A file contains those parts could be | | <p>In the collection, we have secreted mcbABCD to help its colonization in the gut, and secreted CaAP to facilitate calcium uptake by gut cells. We test all parts used in these two circles: 5 signal peptide for secretion, 14 promoters for different strength expression (previously publish and available in <a href="https://addgene.org" target="_blank">addgene.org</a> but not in <a href="http://parts.igem.org/Main_Page" target="_blank">the Registry</a>), IPTG inducible promoter driven various protein coding sequences (confirmed individually before assembled together), and quorum-sensing components. A file contains those parts could be |
| <a href="https://static.igem.org/mediawiki/2020/d/d5/T--Fudan--part_collection_list.xlsx ">downloaded (updated on 2020-10-27)</a>.</p> | | <a href="https://static.igem.org/mediawiki/2020/d/d5/T--Fudan--part_collection_list.xlsx ">downloaded (updated on 2020-10-27)</a>.</p> |
Line 232: |
Line 233: |
| <h2>Five signal peptide for secretion</h2> | | <h2>Five signal peptide for secretion</h2> |
| <div class="column full_size"> | | <div class="column full_size"> |
− | <p>We have constructed plasmids using CaAP or MBP (Maltose Binding Protein, a protein that is expressed in the cytoplasm of <i>E.coli</i>) with five signal peptides (CaAP fused with signal peptides: BBa_K3606031~BBa_K3606035; MBP fused: BBa_K3606037~BBa_K3606041). We have quickly test several of them: the total production and relative amount being secreted. We show that OmpA, one of our signal peptides, could lead MBP to secret. Our results consist with the literature that for a specfic protein all possible signal peptides should be test for best results.</p> | + | <p>We have constructed plasmids using CaAP or MBP (Maltose Binding Protein, a protein that is expressed in the cytoplasm of <i>E. coli</i>) with five signal peptides (CaAP fused with signal peptides: BBa_K3606031~BBa_K3606035; MBP fused: BBa_K3606037~BBa_K3606041). We have quickly test several of them: the total production and relative amount being secreted. We show that OmpA, one of our signal peptides, could lead MBP to secret. Our results consist with the literature that for a specfic protein all possible signal peptides should be test for best results.</p> |
| </div> | | </div> |
| <div class="column half_size"> | | <div class="column half_size"> |
Line 242: |
Line 243: |
| <img src="https://static.igem.org/mediawiki/2020/6/6f/T--Fudan--2020-10-27-gel2.jpg" alt="1027 gel2" /> | | <img src="https://static.igem.org/mediawiki/2020/6/6f/T--Fudan--2020-10-27-gel2.jpg" alt="1027 gel2" /> |
| <h6>Figure 2. IPTG induced signal peptide fused CaAP</h6> | | <h6>Figure 2. IPTG induced signal peptide fused CaAP</h6> |
− | <p>Lane 4 and 5, bacteria DH5a with PhoA-CaAP, culture to OD 0.6; (lane 5) was induced with 1mM IPTG for 6 hours. Then, the bacteria whole-cell samples were run on 17% SDS-PAGE. Lane 1, aprotinin 6.5 kDa.</p> | + | <p>When wiki freeze, we do NOT have Plac driven PhoA-CaAP.<br/>Bacteria DH5a w/ pGEX-6P1, culture to OD 0.6; (lane 5) was induced with 1mM IPTG for 6 hours. Then, the bacteria whole-cell samples were run on 17% SDS-PAGE. Lane 1, aprotinin 6.5 kDa.</p> |
| </div> | | </div> |
| | | |
Line 283: |
Line 284: |
| <div class="column half_size"> | | <div class="column half_size"> |
| <img src="https://static.igem.org/mediawiki/2020/3/31/T--Fudan--2020108-027.jpg" alt="mcbE F G" /> | | <img src="https://static.igem.org/mediawiki/2020/3/31/T--Fudan--2020108-027.jpg" alt="mcbE F G" /> |
− | <h6>Figure 6. IPTG induced mcbE mcbF mcbG</h6> | + | <h6>Figure 6. IPTG induced mcbE mcbF</h6> |
− | <p>Red dot, aprotinin 6.5 kDa; lane 2 vs 1, mcbE induced by IPTG; lane 4 vs 3, mcbF induced by IPTG; lane 6 vs 5, mcbG induced by IPTG. Bacteria were grown to OD 0.6, induced with or without 1 mM IPTG for 5 hours. Whole-cell samples were run. Yellow dots were placed to show induced bands in lane 2,4,6.</p> | + | <p>Red dot, aprotinin 6.5 kDa; lane 2 vs 1, mcbE induced by IPTG; lane 4 vs 3, mcbF induced by IPTG; lane 6 vs 5, GST induced by IPTG. Bacteria were grown to OD 0.6, induced with or without 1 mM IPTG for 5 hours. Whole-cell samples were run. Yellow dots were placed to show induced bands in lane 2,4,6.<br/>When wiki freeze, we do NOT have Plac driven mcbG.</p><!-- lane 6 vs 5, not McbG 21.8 kDa --> |
| </div> | | </div> |
| | | |
| + | <div class="clear"></div> |
| | | |
− | <div class="clear extra_space"></div><!-- 一个章节结束需要clear来留白 如果觉得内容间差异大再加上extra_space --> | + | <div class="column full_size"> |
| + | <p>2020 Team Fudan parts at <a href="http://parts.igem.org/cgi/partsdb/pgroup.cgi?pgroup=iGEM2020&group=Fudan" target="_blank">parts.igem.org</a>, including:</p> |
| + | </div> |
| + | <!-- 本页面特殊的 groupparts 自动生成表格,需要先关掉html再打开 --> |
| + | </html> |
| + | <groupparts>iGEM20 Fudan</groupparts> |
| + | <html> |
| | | |
| <div class="column full_size"><div id="FudanSignature"> | | <div class="column full_size"><div id="FudanSignature"> |
Line 299: |
Line 307: |
| 每次active页面将会由js自动判断 | | 每次active页面将会由js自动判断 |
| --> | | --> |
− | <div class="clear"></div><!-- MUST HAVE -->
| + | |
| + | <div class="clear"></div><!-- MUST HAVE --> |
| </article> | | </article> |
| | | |
Line 307: |
Line 316: |
| <div class="container"><div class="row"> | | <div class="container"><div class="row"> |
| <div class="col m3 s12 row" | | <div class="col m3 s12 row" |
− | style="margin-top:4rem" > | + | style="margin-top:4rem" tabIndex="0" > |
| <a href="http://www.yfc.cn/" target="_blank"><img class="col s3 m6 l4 FudanSponsors" alt="Yunfeng Capital" src="https://static.igem.org/mediawiki/2020/b/ba/T--Fudan--YFcapital.svg"> </a> | | <a href="http://www.yfc.cn/" target="_blank"><img class="col s3 m6 l4 FudanSponsors" alt="Yunfeng Capital" src="https://static.igem.org/mediawiki/2020/b/ba/T--Fudan--YFcapital.svg"> </a> |
| <a href="http://www.fudan.edu.cn/en/" target="_blank"><img alt="Fudan University" class="col s3 m6 l4 FudanSponsors" src="https://static.igem.org/mediawiki/2020/9/9d/T--Fudan--sponsor2.svg"> </a> | | <a href="http://www.fudan.edu.cn/en/" target="_blank"><img alt="Fudan University" class="col s3 m6 l4 FudanSponsors" src="https://static.igem.org/mediawiki/2020/9/9d/T--Fudan--sponsor2.svg"> </a> |
Line 317: |
Line 326: |
| <div class="col s12 l6 row"> | | <div class="col s12 l6 row"> |
| <div class="col s12 m4"> | | <div class="col s12 m4"> |
− | <span><a href="/Team:Fudan/Description#Project">Project</a></span> | + | <span tabIndex="2"><a href="/Team:Fudan/Description#Project">Project</a></span> |
| <ul><li><a href="/Team:Fudan/Description">Description</a></li> | | <ul><li><a href="/Team:Fudan/Description">Description</a></li> |
| <li><a href="/Team:Fudan/Sustainable">Sustainable</a></li> | | <li><a href="/Team:Fudan/Sustainable">Sustainable</a></li> |
Line 325: |
Line 334: |
| </ul></div> | | </ul></div> |
| <div class="col s12 m4"> | | <div class="col s12 m4"> |
− | <span><a href="/Team:Fudan/Notebook">Notebook</a></span> | + | <span tabIndex="3"><a href="/Team:Fudan/Notebook">Notebook</a></span> |
| <ul><li><a href="/Team:Fudan/Safety">Safety</a></li> | | <ul><li><a href="/Team:Fudan/Safety">Safety</a></li> |
| <li><a href="/Team:Fudan/Poster">Poster</a></li> | | <li><a href="/Team:Fudan/Poster">Poster</a></li> |
Line 333: |
Line 342: |
| </ul></div> | | </ul></div> |
| <div class="col s12 m4"> | | <div class="col s12 m4"> |
− | <span><a href="/Team:Fudan/Results">Results</a></span> | + | <span tabIndex="4"><a href="/Team:Fudan/Results">Results</a></span> |
| <ul><li><a href="/Team:Fudan/Contribution">Contribution</a></li><!-- replace Judging --> | | <ul><li><a href="/Team:Fudan/Contribution">Contribution</a></li><!-- replace Judging --> |
| <li><a href="/Team:Fudan/Implementation">Implementation</a></li> | | <li><a href="/Team:Fudan/Implementation">Implementation</a></li> |
Line 343: |
Line 352: |
| <div class="col s12 l6 row"> | | <div class="col s12 l6 row"> |
| <div class="col s12 m4"> | | <div class="col s12 m4"> |
− | <span><a href="/Team:Fudan/Parts">Parts</a></span> | + | <span tabIndex="5"><a href="/Team:Fudan/Parts">Parts</a></span> |
| <ul><li><a href="/Team:Fudan/Parts#Basic">Basic part</a></li> | | <ul><li><a href="/Team:Fudan/Parts#Basic">Basic part</a></li> |
| <li><a href="/Team:Fudan/Parts#Composite">Composite part</a></li> | | <li><a href="/Team:Fudan/Parts#Composite">Composite part</a></li> |
Line 350: |
Line 359: |
| </ul></div> | | </ul></div> |
| <div class="col s12 m4"> | | <div class="col s12 m4"> |
− | <span><a href="/Team:Fudan/Human_Practices">Outreach</a></span> | + | <span tabIndex="6"><a href="/Team:Fudan/Human_Practices">Outreach</a></span> |
| <ul><li><a href="/Team:Fudan/Human_Practices#Integrated">Integrated HP</a></li> | | <ul><li><a href="/Team:Fudan/Human_Practices#Integrated">Integrated HP</a></li> |
| <li><a href="/Team:Fudan/Education">Education</a></li> | | <li><a href="/Team:Fudan/Education">Education</a></li> |
Line 357: |
Line 366: |
| </ul></div> | | </ul></div> |
| <div class="col s12 m4"> | | <div class="col s12 m4"> |
− | <span><a href="/Team:Fudan/Team">Team</a></span> | + | <span tabIndex="7"><a href="/Team:Fudan/Team">Team</a></span> |
| <ul><li><a href="/Team:Fudan/Team">Members</a></li> | | <ul><li><a href="/Team:Fudan/Team">Members</a></li> |
| <li><a href="/Team:Fudan/Attributions">Attributions</a></li> | | <li><a href="/Team:Fudan/Attributions">Attributions</a></li> |
| <li><a href="/Team:Fudan/Partnership">Partnership</a></li> | | <li><a href="/Team:Fudan/Partnership">Partnership</a></li> |
| <li><a href="/Team:Fudan/Acknowledgement">Acknowledgement</a></li> | | <li><a href="/Team:Fudan/Acknowledgement">Acknowledgement</a></li> |
− | <li><a href="https://2019.igem.org/Team:Fudan-TSI/Heritage" target="_blank">Heritage</a></li> | + | <li><a href="/Team:Fudan/Heritage">Heritage</a></li> |
| </ul></div> | | </ul></div> |
| <div class="col s12 m4"> </div> | | <div class="col s12 m4"> </div> |