Difference between revisions of "Team:Tokyo Tech/Basic Part"

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{{Tokyo_Tech}}
 
{{Tokyo_Tech}}
  
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<h1 align="center">Parts Basic Parts</h1>
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<div id="best_basic_part" class="container">
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<div id="bset_basic_part_header" class="container_header">
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<h1><span>Best Basic part: BBa_K1949050</span>
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</div><!-- /best_basic_part_header -->
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<div id="best_basic_part_contents" class="container_contents">
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<h2 align="center">BBa_K1949050 meets the Silver Medal criteria!</h2>
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<p class="normal_text">The Prince <span style="font-style: italic;">coli</span> expresses AmiE protein, and Snow White coli recovers from its apparent death and wakes up again. We tested the function of AmiE protein that influences the end of the story. </p>
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<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 is not degraded by AmiE, but 3OC12HSL is degraded by AmiE.</p>
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<ul>
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<li><p class="normal_text">PBAD/araC-<span style="font-style: italic;">rbs-amiE</span>(pSB6A1)</p></li>
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<li><p class="normal_text">Ptet-<span style="font-style: italic;">rbs-luxR</span>-<span style="font-style: italic;">tt</span>-Plux-<span style="font-style: italic;">rbs-gfp</span> (pSB6A1)</p></li>
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<li><p class="normal_text">Ptet-<span style="font-style: italic;">rbs-luxR</span>-<span style="font-style: italic;">tt</span>-Plux-<span style="font-style: italic;">rbs-gfp</span> (pSB6A1)</p></li>
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</ul>
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<div id="other_basic_part" class="container">
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<div id="other_basic_part_header" class="container_header">
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<h1><span>Tokyo Tech 2015 iGEM Team: The Others Basic Parts</span></h1>
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<table class="parts_table">
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<tr>
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<th class="parts_table_name">Name</th>
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<th class="parts_table_type">Type</th>
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<th class="parts_table_description">Description</th>
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<th class="parts_table_design">Design</th>
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<th class="parts_table_length">Length(bp)</th>
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</tr>
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<tr>
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<td><a href="http://parts.igem.org/Part:BBa_K1949000">BBa_K1949000</a></td>
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<td>Regulatory</td>
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<td>Pcold</td>
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<td>Yoshio Takata</td>
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<td>313</td>
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</tr>
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<tr>
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<td><a href="http://parts.igem.org/Part:BBa_K1949020">BBa_K1949020</a></td>
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<td>Coding</td>
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<td><span style="font-style:italic;">yafN</span></td>
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<td>Kazuki Fujisawa</td>
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<td>297</td>
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</tr>
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<td><a href="http://parts.igem.org/Part:BBa_K1949030">BBa_K1949030</a></td>
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<td>Coding</td>
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<td><span style="font-style:italic;">yafO</span></td>
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<td>Yoshio Takata</td>
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<td>402</td>
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</tr>
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<tr>
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<td><a href="http://parts.igem.org/Part:BBa_K1949031">BBa_K1949031</a></td>
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<td>Transrational unit</td>
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<td><span style="font-style:italic;">rbs</span>-<span style="font-style:italic;">yafO</span></td>
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<td>Yoshio Takata</td>
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<td>420</td>
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</tr>
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<tr>
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<td><a href="http://parts.igem.org/Part:BBa_K1949050">BBa_K1949050</a></td>
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<td>Coding</td>
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<td><span style="font-style:italic;">amiE</span></td>
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<td>Yoshio Takata</td>
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<td>1476</td>
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</tr>
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<tr>
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<td><a href="http://parts.igem.org/Part:BBa_K1949051">BBa_K1949051</a></td>
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<td>Transrational unit</td>
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<td><span style="font-style:italic;">rbs-amiE</span></td>
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<td>Yoshio Takata</td>
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<td>1494</td>
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</tr>
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</tbody>
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</table>
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</div><!-- /other_basic_part_contents -->
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</div><!-- /other_basic_part -->
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<div id="pcold" class="container">
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<div id="pcold_header" class="container_header">
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<h1><span>Cold inducible promoter (Pcold): <a href="http://parts.igem.org/Part:BBa_K1949000">BBa_K1949000</a></h1>
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</div><!-- /pcold_header -->
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<div id="pcold_contents" class="container_contents">
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<h2 align="center"><a href="http://parts.igem.org/Part:BBa_K1949000">BBa_K1949000</a> meets the Bronze Medal criteria!</h2>
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<p class="normal_text"> This promoter is used to effectively produce proteins at low temperatures. This new promoter, a cold inducible promoter (we call this Pcold) consists of the <span style="font-style: italic; ">cspA</span> promoter, Cold Box, 5’-UTR, RBS and DB. The combination (of <span style="font-style: italic; ">cspA</span> promoter which is active at both low and high temperature, Cold box which inhibits excessive gene expression, 5’UTR which is stable at only low temperature, and DB which function as an extra RBS) activates gene expression at low temperatures.</p>
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<p class="normal_text">RFU (Relative Fluorescence Units) of GFP per turbidity was measured using cells cultured at 18℃ and 37℃ to confirm function of Pcold. The cells harbored a plasmid which carries Pcold-<span style="font-style: italic; ">gfp</span> or Ptet-<span style="font-style: italic; ">rbs-gfp</span>.</p>
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<p class="normal_text"><span style="font-style: italic; ">E. coli</span> cells which carry the Ptet-<span style="font-style: italic; ">rbs-gfp</span> plasmid was cultured at 18°C, and RFU of GFP was measured at indicated time points. Also, the same experiment was performed at 37°C. We thought this result was obtained because GFP is easily folded into correct structures at low temperatures. By contrast, RFU of <span style="font-style: italic; ">E. coli</span> which harbored Pcold-<span style="font-style: italic; ">rbs-gfp</span> at 18°C was about eight fold higher than that at 37°C. From this result, we confirmed Pcold activates gene expression at low temperatures.</p>
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</div><!-- /pcold_contents -->
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</div><!-- /main_contents -->
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</body>
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{{Tokyo_Tech/Footer}}
 
{{Tokyo_Tech/Footer}}

Revision as of 11:56, 18 October 2016

Parts Basic Parts

Best Basic part: BBa_K1949050

BBa_K1949050 meets the Silver Medal criteria!

The Prince coli expresses AmiE protein, and Snow White coli recovers from its apparent death 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 is not degraded by AmiE, but 3OC12HSL is degraded by AmiE.

  • PBAD/araC-rbs-amiE(pSB6A1)

  • Ptet-rbs-luxR-tt-Plux-rbs-gfp (pSB6A1)

  • Ptet-rbs-luxR-tt-Plux-rbs-gfp (pSB6A1)

Tokyo Tech 2015 iGEM Team: The Others Basic Parts

Name Type Description Design Length(bp)
BBa_K1949000 Regulatory Pcold Yoshio Takata 313
BBa_K1949020 Coding yafN Kazuki Fujisawa 297
BBa_K1949030 Coding yafO Yoshio Takata 402
BBa_K1949031 Transrational unit rbs-yafO Yoshio Takata 420
BBa_K1949050 Coding amiE Yoshio Takata 1476
BBa_K1949051 Transrational unit rbs-amiE Yoshio Takata 1494

Cold inducible promoter (Pcold): BBa_K1949000

BBa_K1949000 meets the Bronze Medal criteria!

This promoter is used to effectively produce proteins at low temperatures. This new promoter, a cold inducible promoter (we call this Pcold) consists of the cspA promoter, Cold Box, 5’-UTR, RBS and DB. The combination (of cspA promoter which is active at both low and high temperature, Cold box which inhibits excessive gene expression, 5’UTR which is stable at only low temperature, and DB which function as an extra RBS) activates gene expression at low temperatures.

RFU (Relative Fluorescence Units) of GFP per turbidity was measured using cells cultured at 18℃ and 37℃ to confirm function of Pcold. The cells harbored a plasmid which carries Pcold-gfp or Ptet-rbs-gfp.

E. coli cells which carry the Ptet-rbs-gfp plasmid was cultured at 18°C, and RFU of GFP was measured at indicated time points. Also, the same experiment was performed at 37°C. We thought this result was obtained because GFP is easily folded into correct structures at low temperatures. By contrast, RFU of E. coli which harbored Pcold-rbs-gfp at 18°C was about eight fold higher than that at 37°C. From this result, we confirmed Pcold activates gene expression at low temperatures.