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+ | <head> | ||
+ | <meta charset="UTF-8" /> | ||
+ | <style type="text/css"> | ||
+ | <!-- | ||
+ | /********************** All style of this page ************************/ | ||
+ | body{ | ||
+ | background-image: url("https://static.igem.org/mediawiki/2016/e/eb/T--Tokyo_Tech--Background.png"); | ||
+ | } | ||
+ | #globalWrapper, #content{ | ||
+ | background: transparent; | ||
+ | } | ||
+ | #content{ | ||
+ | // background-image: url("https://static.igem.org/mediawiki/2016/e/eb/T--Tokyo_Tech--Background.png"); | ||
+ | margin: 0 auto; | ||
+ | } | ||
+ | /**********************************************************************/ | ||
+ | #main_contents{ | ||
+ | margin: 0 auto; | ||
+ | width: 90%; | ||
+ | min-width: 880px; | ||
+ | max-width: 1100px; | ||
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+ | line-height: 1.5em; | ||
+ | width: 800px; | ||
+ | margin: 0 auto; | ||
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+ | } | ||
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+ | font-size: 18px; | ||
+ | text-indent: 18px; | ||
+ | text-align: justify; | ||
+ | // padding: 18px; | ||
+ | margin: 0; | ||
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+ | color: #0000000; | ||
+ | line-height: 1.5em; | ||
+ | } | ||
+ | #main_contents table th{ | ||
+ | font-size: 20px; | ||
+ | vertical-align: middle; | ||
+ | } | ||
+ | |||
+ | /************************* this page setting ******************/ | ||
+ | #main_contents a{ | ||
+ | text-decoration: none; | ||
+ | } | ||
+ | #main_contents .parts_table{ | ||
+ | width: 800px; | ||
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+ | } | ||
+ | #main_contents td, #main_contents th{ | ||
+ | border: 1px solid #000000; | ||
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+ | #main_contents th.parts_table_name, #main_contents th.parts_table_type { | ||
+ | width: 150px; | ||
+ | } | ||
+ | #main_contents th.parts_table_design{ | ||
+ | width: 120px; | ||
+ | }r | ||
+ | #main_contents th.parts_table_description{ | ||
+ | width: 220px; | ||
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+ | #main_contens th.parts_table_length{ | ||
+ | width: auto; | ||
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+ | --></style> | ||
+ | </head> | ||
<body> | <body> | ||
+ | <div id="main_contents"> | ||
+ | <div id="page_header" class="container container_top"> | ||
+ | <h1 align="center">Parts</h1> | ||
+ | </div><!-- /page_header --> | ||
+ | <div id="parts_overview" class="container"> | ||
+ | <div id="parts_overview_contents" class="container_contents"> | ||
+ | <p class="normal_text"><b>To meet the criteria of the Gold Medal</b>, we submitted <a href="http://parts.igem.org/Part:BBa_K1949060">BBa_K1949060</a>
and we characterized <a href="http://parts.igem.org/Part:BBa_R0071">BBa_R0071</a>, <a href="http://parts.igem.org/Part:BBa_C0071">BBa_C0071</a>, <a href="http://parts.igem.org/Part:BBa_C0171">BBa_C0171</a>, <a href="http://parts.igem.org/Part:BBa_K1529300">BBa_K1529300</a>, <a href="http://parts.igem.org/Part:BBa_K1529310">BBa_K1529310</a>, <a href="http://parts.igem.org/Part:BBa_K1096001">BBa_K1096001</a> and <a href="http://parts.igem.org/Part:BBa_K1096002">BBa_K1096002</a>. </p> | ||
+ | <p class="normal_text"><b>To meet the criteria of the Silver Medal</b>, we submitted <a href="http://parts.igem.org/Part:BBa_K1949050">BBa_K1949050</a>, <a href="http://parts.igem.org/Part:BBa_K1949052">BBa_K1949052</a>, <a href="http://parts.igem.org/Part:BBa_K1949030">BBa_K1949030</a> and <a href="http://parts.igem.org/Part:BBa_K1949032">BBa_K1949032</a>.</p> | ||
+ | <p class="normal_text"><b>To meet the criteria of the Bronze Medal</b>, we submitted <a href="http://parts.igem.org/Part:BBa_K1949000">BBa_K1949000</a>, | ||
+ | <a href="http://parts.igem.org/Part:BBa_K1949001">BBa_K1949001</a>, <a href="http://parts.igem.org/Part:BBa_K1949100">BBa_K1949100</a>, <a href="http://parts.igem.org/Part:BBa_K1949101">BBa_K1949101</a> and <a href="http://parts.igem.org/Part:BBa_K1949102">BBa_K1949102</a>.</p> | ||
+ | </div><!-- /parts_overview_contents --> | ||
+ | </div><!-- /parts_overview --> | ||
+ | |||
+ | <div id="parts_2016" class="container"> | ||
+ | <div id="favorite_parts_header" class="container_header"> | ||
+ | <h1><span>Favorite Tokyo Tech 2016 iGEM Team Parts</span></h1> | ||
+ | </div><!-- /favorite_parts_header --> | ||
+ | <div id="favorite_parts_contents" class="container_contents"> | ||
+ | <table border="1" align="center" class="parts_table"> | ||
+ | <tbody> | ||
+ | <tr> | ||
+ | <th class="parts_table_name">Name</th> | ||
+ | <th class="parts_table_type">Type</th> | ||
+ | <th class="parts_table_description">Description</th> | ||
+ | <th class="parts_table_design">Design</th> | ||
+ | <th class="parts_table_length">Length(bp)</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td><a href="http://parts.igem.org/Part:BBa_K1949050">BBa_K1949050</a></td> | ||
+ | <td>Coding</td> | ||
+ | <td><span style="font-style:italic;">amiE</span></td> | ||
+ | <td>Yoshio Takata</td> | ||
+ | <td>1476</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td><a href="http://parts.igem.org/Part:BBa_K1949060">BBa_K1949060</a></td> | ||
+ | <td>Composite</td> | ||
+ | <td>Prhl(NM)-<span style="font-style:italic;">rbs</span>-<span style="font-style:italic;">gfp</span></td> | ||
+ | <td>Yoshio Takata</td> | ||
+ | <td>799</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td><a href="http://parts.igem.org/Part:BBa_K1949102">BBa_K1949102</a></td> | ||
+ | <td>Composite</td> | ||
+ | <td>PBAD-<span style="font-style:italic;">rbs</span>-<span style="font-style:italic;">mazF</span>-<span style="font-style:italic;">tt</span>-Ptet-<span style="font-style:italic;">rbs</span>-<span style="font-style:italic;">gfp</span></td> | ||
+ | <td>Yoshio Takata</td> | ||
+ | <td>2520</td> | ||
+ | </tr> | ||
+ | </tbody> | ||
+ | </table> | ||
+ | </div><!-- /favorite_parts_contents --> | ||
+ | |||
+ | <div id="normal_parts_header" class="container_header"> | ||
+ | <h1><span>Tokyo Tech 2016 iGEM Team Parts</span></h1> | ||
+ | </div><!-- /normal_parts_header --> | ||
+ | <div id="normal_parts_contents" class="container_contents"> | ||
+ | <table border="1" align="center" class="parts_table"> | ||
+ | <tbody> | ||
+ | <tr> | ||
+ | <th class="parts_table_name">Name</th> | ||
+ | <th class="parts_table_type">Type</th> | ||
+ | <th class="parts_table_description">Description</th> | ||
+ | <th class="parts_table_design">Design</th> | ||
+ | <th class="parts_table_length">Length(bp)</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td><a href="http://parts.igem.org/Part:BBa_K1949000">BBa_K1949000</a></td> | ||
+ | <td>Regulatory</td> | ||
+ | <td>Pcold</td> | ||
+ | <td>Yoshio Takata</td> | ||
+ | <td>313</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td><a href="http://parts.igem.org/Part:BBa_K1949001">BBa_K1949001</a></td> | ||
+ | <td>Measurement</td> | ||
+ | <td>Pcold-<span style="font-style:italic;">gfp</span></td> | ||
+ | <td>Yoshio Takata</td> | ||
+ | <td>1033</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td><a href="http://parts.igem.org/Part:BBa_K1949020">BBa_K1949020</a></td> | ||
+ | <td>Coding</td> | ||
+ | <td><span style="font-style:italic;">yafN</span></td> | ||
+ | <td>Kazuki Fujisawa</td> | ||
+ | <td>297</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td><a href="http://parts.igem.org/Part:BBa_K1949022">BBa_K1949022</a></td> | ||
+ | <td>Composite</td> | ||
+ | <td>Plac-<span style="font-style:italic;">rbs</span>-<span style="font-style:italic;">yafN</span<</td> | ||
+ | <td>Kazuki Fujisawa</td> | ||
+ | <td>523</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td><a href="http://parts.igem.org/Part:BBa_K1949030">BBa_K1949030</a></td> | ||
+ | <td>Coding</td> | ||
+ | <td><span style="font-style:italic;">yafO</span></td> | ||
+ | <td>Yoshio Takata</td> | ||
+ | <td>402</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td><a href="http://parts.igem.org/Part:BBa_K1949031">BBa_K1949031</a></td> | ||
+ | <td>Transrational unit</td> | ||
+ | <td><span style="font-style:italic;">rbs</span>-<span style="font-style:italic;">yafO</span></td> | ||
+ | <td>Yoshio Takata</td> | ||
+ | <td>420</td> | ||
+ | </tr> | ||
+ | |||
+ | <tr> | ||
+ | <td><a href="http://parts.igem.org/Part:BBa_K1949032">BBa_K1949032</a></td> | ||
+ | <td>Composite</td> | ||
+ | <td>PBAD-<span style="font-style:italic;">rbs-yafO</span></td> | ||
+ | <td>Yoshio Takata</td> | ||
+ | <td>1638</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td><a href="http://parts.igem.org/Part:BBa_K1949033">BBa_K1949033</a></td> | ||
+ | <td>Composite</td> | ||
+ | <td>PBAD-<span style="font-style:italic;">rbs-yafO-tt</span>-Ptet-<span style="font-style:italic;">rbs-gfp</a></td> | ||
+ | <td>Yoshio Takata</td> | ||
+ | <td>2580</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td><a href="http://parts.igem.org/Part:BBa_K1949051">BBa_K1949051</a></td> | ||
+ | <td>Transrational unit</td> | ||
+ | <td><span style="font-style:italic;">rbs-amiE</span></td> | ||
+ | <td>Yoshio Takata</td> | ||
+ | <td>1494</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td><a href="http://parts.igem.org/Part:BBa_K1949052">BBa_K1949052</a></td> | ||
+ | <td>Composite</td> | ||
+ | <td>PBAD-<span style="font-style:italic;">rbs-amiE</a></td> | ||
+ | <td>Yoshio Takata</td> | ||
+ | <td>2712</td> | ||
+ | </tr> | ||
+ | |||
+ | <tr> | ||
+ | <td><a href="http://parts.igem.org/Part:BBa_K1949100">BBa_K1949100</a></td> | ||
+ | <td>Composite</td> | ||
+ | <td>Plac-<span style="font-style:italic;">rbs-mazE</span></td> | ||
+ | <td>Yoshio Takata</td> | ||
+ | <td>565</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td><a href="http://parts.igem.org/Part:BBa_K1949101">BBa_K1949101</a></td> | ||
+ | <td>Composite</td> | ||
+ | <td>PBAD-<span style="font-style:italic;">rbs-mazF</span></td> | ||
+ | <td>Yoshio Takata</td> | ||
+ | <td>1575</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td><a href="http://parts.igem.org/Part:BBa_K1949103">BBa_K1949103</a></td> | ||
+ | <td>Composite</td> | ||
+ | <td>Ptet-<span style="font-style:italic;">rbs</span>(<a href="http://parts.igem.org/Part:BBa_B0034">BBa_B0034)</a>)-<span style="font-style:italic;">mazE</span></td> | ||
+ | <td>Yoshio Takata</td> | ||
+ | <td>332</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td><a href="http://parts.igem.org/Part:BBa_K1949104">BBa_K1949104</a></td> | ||
+ | <td>Composite</td> | ||
+ | <td>Ptet-<span style="font-style:italic;">rbs</span>(<a href="http://parts.igem.org/Part:BBa_J61117">BBa_J61117</a>)-<span style="font-style:italic;">mazE</span></td> | ||
+ | <td>Yoshio Takata</td> | ||
+ | <td>332</td> | ||
+ | </tr> | ||
+ | </tbody> | ||
+ | </table> | ||
+ | </div><!-- /normal_parts_contents --> | ||
+ | </div><!-- /pats_2016 --> | ||
+ | |||
+ | |||
+ | |||
+ | <div id="improved_part" class="container"> | ||
+ | <div id="improved_part_header" class="container_header"> | ||
+ | <h1><span>1. Improved Part: <a href="http://parts.igem.org/Part:BBa_K1949060">BBa_K1949060</a></span></h1> | ||
+ | </div><!-- /improved_part_header --> | ||
+ | <div id="improved_part_contents" class="container_contents"> | ||
+ | <h2 align="center"><a href="http://parts.igem.org/Part:BBa_K1949060">BBa_K1949060</a> meets the Gold Medal criteria!</h2> | ||
+ | <p class="normal_text"> We simulated our final genetic circuits and found that the circuits would not work, because compared with Plux, Prhl activity was too weak. (see <a href="https://2016.igem.org/Team:Tokyo_Tech/Model"> the Model page </a> and <a href="https://2016.igem.org/Team:Tokyo_Tech/AHL_Assay/Only_Assay">the AHL only assay page</a>). We therefore considered using the improved Prhl (<a href="http://parts.igem.org/Part:BBa_K1529310">BBa_K1529310</a>, <a href="http://parts.igem.org/Part:BBa_K1529300">BBa_K1529300</a>) established by iGEM 2014 Tokyo_Tech team, but we noticed it was inappropriate for two reasons (see <a href="https://2016.igem.org/Team:Tokyo_Tech/AHL_Assay/Rhl_System_Assay">Rhl_system_assay</a>). Then, we decided to further improve this Prhl and obtain our original improved Prhl (Noticeable Mutant) (<a href="http://parts.igem.org/Part:BBa_K1949060">BBa_K1949060</a>). We referred to as Prhl(NM) from now on that suited our goal.</p> | ||
+ | <p class="normal_text">Our purpose is to create a strong Prhl for our final genetic circuits.</p> | ||
+ | <p class="normal_text">This experiment consists of the three parts below. </p> | ||
+ | <ol type="1"> | ||
+ | <li><p class="normal_text">Improved Prhl by iGEM 2014 Tokyo_Tech team and characterization of <i>rhlR</i>(Fig. 4-1-1).</p></li> | ||
+ | <div align="center"><img src="https://static.igem.org/mediawiki/2016/c/cc/T--Tokyo_tech--Fig.4_1_1.png" height="300"><br></div> | ||
+ | <div align="center"><p class="caption" style="font-size: 16px; text-align: center;"><span style="font-weight: bold;">Fig. 4-1-1 Effect of RhlR by tagging LVA <br>and The colonies of transformants with a rhlR (left) or a rhlR - LVA (right)</span> | ||
+ | <li><p class="normal_text">Improvement of the wild type Prhl(Fig. 4-1-2).</p></li> | ||
+ | <div align="center"><img src="https://static.igem.org/mediawiki/2016/a/a7/T--Tokyo_tech--Fig.4_1_2.png" height="300"><br></div> | ||
+ | <div align="center"><p class="caption" style="font-size: 16px; text-align: center;"><span style="font-weight: bold;">Fig. 4-1-2 RFU of GFP / Turbidity of imoroved Prhl mutants </span> | ||
+ | <li><p class="normal_text">Comparison of the improved Prhl by iGEM 2014 Tokyo_Tech team with our original improved Prhl(Fig. 4-1-3)</p></li> | ||
+ | <div align="center"><img src="https://static.igem.org/mediawiki/2016/b/b1/T--Tokyo_tech--Fig.4_1_3.png" height="300"><br></div> | ||
+ | <div align="center"><p class="caption" style="font-size: 16px; text-align: center;"><span style="font-weight: bold;">Fig. 4-1-3 Comparison of Prhl(NM) by Prhl(LR)</span> | ||
+ | </ol> | ||
+ | |||
+ | <p class="normal_text">The past improved Prhl did not suit for our final circuits and we could construct the improved Prhl appropriate to our final circuits.</p> | ||
+ | <p class="normal_text">For more information, read <a href="http://parts.igem.org/Part:BBa_K1949060">this page</a> !</p> | ||
+ | </div><!-- /improved_part_contents --> | ||
+ | </div><!-- /improved_part --> | ||
+ | |||
+ | |||
+ | |||
+ | |||
+ | <div id="best_basic_part" class="container"> | ||
+ | <div id="best_basic_part_header" class="container_header"> | ||
+ | <h1><span>2.Best Basic Part: <a href="http://parts.igem.org/Part:BBa_K1949050">BBa_K1949050</span></h1> | ||
+ | </div><!-- /best_basic_part_header --> | ||
+ | |||
+ | <div id="best_basic_part_contents" class="container_contents"> | ||
+ | <h2 align="center"><a href="http://parts.igem.org/Part:BBa_K1949050">BBa_K1949050</a> meets the Silver Medal criteria!</h2> | ||
+ | |||
+ | <p class="normal_text"><i>amiE</i> codes for protein AmiE. AmiE is an acylase that degrades long chain [N]-acyl homoserine lactone (AHL) molecules with acyl chains longer than six carbons. Prince <span style="font-style:italic;">coli</span>(Fig. 4-1-4) expresses <span style="font-style:italic;">amiE</span>, and Snow White <span style="font-style:italic;">coli</span> resuscitate from its dormancy and wakes up again. We tested the function of AmiE protein that influences the end of the story. </p> | ||
+ | <div align="center"><img src="https://static.igem.org/mediawiki/2016/4/4a/T--Tokyo_tech--Fig.4_2_3%282%29.png" height="200"><br></div> | ||
+ | <div align="center"><p class="caption" style="font-size: 16px; text-align: center;"><span style="font-weight: bold;">Fig. 4-1-4 AmiE degrades long chain [N]-acyl homoserine lactone (AHL) molecules<br> with acyl chains longer than six carbons</span></p></div> | ||
+ | </ol> | ||
+ | <p class="normal_text">Our objective is to characterize the function of AmiE protein. We prepared three samples shown below. When we tested the AmiE degradation ability with these samples, the results show that C4HSL(refer to it as C4 hereafter) is not degraded by AmiE, but 3OC12HSL(C12) is degraded by AmiE(Fig. 4-1-5).</p> | ||
+ | |||
+ | <ul> | ||
+ | <li><p class="normal_text">PBAD ‐ <span style="font-style:italic;">rbs</span> ‐ <span style="font-style:italic;">amiE</span> (pSB6A1)</p></li> | ||
+ | <li><p class="normal_text">Ptet ‐ <span style="font-style:italic;">rbs</span> ‐ <span style="font-style:italic;">luxR ‐ tt</span> ‐ Plux ‐ <span style="font-style:italic;">rbs</span> ‐ <span style="font-style:italic;">gfp</span> (pSB6A1)</p></li> | ||
+ | <li><p class="normal_text">Ptet ‐ <span style="font-style:italic;">rbs ‐ rhlR ‐ tt</span> ‐ Prhl ‐ <span style="font-style:italic;">rbs ‐ gfp</span> (pSB6A1)</p></li> | ||
+ | <div align="center"><img src="https://static.igem.org/mediawiki/2016/8/83/T--Tokyo_tech--cold.png" height="300"><br></div> | ||
+ | <div align="center"><p class="caption" style="font-size: 16px; text-align: center;"><span style="font-weight: bold;">Fig. 4-1-5 AmiE degrades C12 but barely degrades C4</span></p></div> | ||
+ | <p class="normal_text">For more information, read <a href="http://parts.igem.org/Part:BBa_K1949050">this page</a> !</p> | ||
+ | </ul> | ||
+ | </div><!-- /best_basic_part_contents --> | ||
+ | </div><!-- /best_basic_part --> | ||
+ | |||
+ | |||
+ | <div id="best_composite_part" class="container"> | ||
+ | <div id="best_composite_part_header" class="container_header"> | ||
+ | <h1><span>Best Composite Part: <a href="http://parts.igem.org/Part:BBa_K1949102">BBa_K1949102</a></span></h1> | ||
+ | </div><!-- /best_composite_part_header --> | ||
+ | |||
+ | <div id="bast_composite_part_contents" class="container_contents"> | ||
+ | <h2 align="center"><a href="http://parts.igem.org/Part:BBa_K1949102">BBa_K1949102</a> meets the Bronze Medal criteria!</h2> | ||
+ | |||
+ | <p class="normal_text">The biggest attraction of the TA system is that it is able to control cell growth and synthesis of protein. In this experiment, <span style="font-style: italic;">mazE</span> expression was induced by the addition of IPTG (2 mM) after <span style="font-style: italic;">mazF</span> expression was induced by the addition of arabinose(0.02%). As a result, it was able to resuscitate from a state of being inhibited cell growth. We named this experiment as "Stop & Go" because it was to resuscitate growth from inhibiting cell growth.</p> | ||
+ | |||
+ | <div align="center"><img src="https://static.igem.org/mediawiki/2016/7/77/T--Tokyo_tech--Fig.4.png" height="200"><br></div> | ||
+ | <div align="center"><p class="caption" style="font-size: 16px; text-align: center;"><span style="font-weight: bold;">Fig. 4-1-6 Time vs Turbidity (Stop & Go) and Time vs RFU of GFP (Stop & Go)</span> | ||
+ | |||
+ | <p class="normal_text">It was found from Fig. 4-1-6 that MazF inhibited cell growth. MazE was induced 2 h after <span style="font-style: italic;">mazE</span> expression, and about 8 h later, cell growth which had stopped was resuscitated. From these results, it was suggested that <span style="font-style: italic;">E. coli</span> whose cell growth was inhibited by MazF was able to resuscitate by expression of <span style="font-style: italic;">mazE</span>. | ||
+ | <p class="normal_text">For more information, read <a href="http://parts.igem.org/Part:BBa_K1949102">this page</a> !</p> | ||
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Latest revision as of 02:20, 20 October 2016
Parts
To meet the criteria of the Gold Medal, we submitted BBa_K1949060 and we characterized BBa_R0071, BBa_C0071, BBa_C0171, BBa_K1529300, BBa_K1529310, BBa_K1096001 and BBa_K1096002.
To meet the criteria of the Silver Medal, we submitted BBa_K1949050, BBa_K1949052, BBa_K1949030 and BBa_K1949032.
To meet the criteria of the Bronze Medal, we submitted BBa_K1949000, BBa_K1949001, BBa_K1949100, BBa_K1949101 and BBa_K1949102.
Favorite Tokyo Tech 2016 iGEM Team Parts
Name | Type | Description | Design | Length(bp) |
---|---|---|---|---|
BBa_K1949050 | Coding | amiE | Yoshio Takata | 1476 |
BBa_K1949060 | Composite | Prhl(NM)-rbs-gfp | Yoshio Takata | 799 |
BBa_K1949102 | Composite | PBAD-rbs-mazF-tt-Ptet-rbs-gfp | Yoshio Takata | 2520 |
Tokyo Tech 2016 iGEM Team Parts
Name | Type | Description | Design | Length(bp) |
---|---|---|---|---|
BBa_K1949000 | Regulatory | Pcold | Yoshio Takata | 313 |
BBa_K1949001 | Measurement | Pcold-gfp | Yoshio Takata | 1033 |
BBa_K1949020 | Coding | yafN | Kazuki Fujisawa | 297 |
BBa_K1949022 | Composite | Plac-rbs-yafN | Kazuki Fujisawa | 523 |
BBa_K1949030 | Coding | yafO | Yoshio Takata | 402 |
BBa_K1949031 | Transrational unit | rbs-yafO | Yoshio Takata | 420 |
BBa_K1949032 | Composite | PBAD-rbs-yafO | Yoshio Takata | 1638 |
BBa_K1949033 | Composite | PBAD-rbs-yafO-tt-Ptet-rbs-gfp | Yoshio Takata | 2580 |
BBa_K1949051 | Transrational unit | rbs-amiE | Yoshio Takata | 1494 |
BBa_K1949052 | Composite | PBAD-rbs-amiE | Yoshio Takata | 2712 |
BBa_K1949100 | Composite | Plac-rbs-mazE | Yoshio Takata | 565 |
BBa_K1949101 | Composite | PBAD-rbs-mazF | Yoshio Takata | 1575 |
BBa_K1949103 | Composite | Ptet-rbs(BBa_B0034))-mazE | Yoshio Takata | 332 |
BBa_K1949104 | Composite | Ptet-rbs(BBa_J61117)-mazE | Yoshio Takata | 332 |
1. Improved Part: BBa_K1949060
BBa_K1949060 meets the Gold Medal criteria!
We simulated our final genetic circuits and found that the circuits would not work, because compared with Plux, Prhl activity was too weak. (see the Model page and the AHL only assay page). We therefore considered using the improved Prhl (BBa_K1529310, BBa_K1529300) established by iGEM 2014 Tokyo_Tech team, but we noticed it was inappropriate for two reasons (see Rhl_system_assay). Then, we decided to further improve this Prhl and obtain our original improved Prhl (Noticeable Mutant) (BBa_K1949060). We referred to as Prhl(NM) from now on that suited our goal.
Our purpose is to create a strong Prhl for our final genetic circuits.
This experiment consists of the three parts below.
Improved Prhl by iGEM 2014 Tokyo_Tech team and characterization of rhlR(Fig. 4-1-1).
Improvement of the wild type Prhl(Fig. 4-1-2).
Comparison of the improved Prhl by iGEM 2014 Tokyo_Tech team with our original improved Prhl(Fig. 4-1-3)
PBAD ‐ rbs ‐ amiE (pSB6A1)
Ptet ‐ rbs ‐ luxR ‐ tt ‐ Plux ‐ rbs ‐ gfp (pSB6A1)
Ptet ‐ rbs ‐ rhlR ‐ tt ‐ Prhl ‐ rbs ‐ gfp (pSB6A1)
The past improved Prhl did not suit for our final circuits and we could construct the improved Prhl appropriate to our final circuits.
For more information, read this page !
2.Best Basic Part: BBa_K1949050
BBa_K1949050 meets the Silver Medal criteria!
amiE codes for protein AmiE. AmiE is an acylase that degrades long chain [N]-acyl homoserine lactone (AHL) molecules with acyl chains longer than six carbons. Prince coli(Fig. 4-1-4) expresses amiE, and Snow White coli resuscitate from its dormancy and wakes up again. We tested the function of AmiE protein that influences the end of the story.
Our objective is to characterize the function of AmiE protein. We prepared three samples shown below. When we tested the AmiE degradation ability with these samples, the results show that C4HSL(refer to it as C4 hereafter) is not degraded by AmiE, but 3OC12HSL(C12) is degraded by AmiE(Fig. 4-1-5).
For more information, read this page !
Best Composite Part: BBa_K1949102
BBa_K1949102 meets the Bronze Medal criteria!
The biggest attraction of the TA system is that it is able to control cell growth and synthesis of protein. In this experiment, mazE expression was induced by the addition of IPTG (2 mM) after mazF expression was induced by the addition of arabinose(0.02%). As a result, it was able to resuscitate from a state of being inhibited cell growth. We named this experiment as "Stop & Go" because it was to resuscitate growth from inhibiting cell growth.
It was found from Fig. 4-1-6 that MazF inhibited cell growth. MazE was induced 2 h after mazE expression, and about 8 h later, cell growth which had stopped was resuscitated. From these results, it was suggested that E. coli whose cell growth was inhibited by MazF was able to resuscitate by expression of mazE.
For more information, read this page !