Difference between revisions of "Team:Tokyo Tech/Parts"

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<p class="normal_text">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. </p>
 
<p class="normal_text">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. </p>
 +
                              <div align="center"><img src="https://static.igem.org/mediawiki/2016/a/a7/T--Tokyo_tech--Fig.4_1_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. 4-1-4</span>
 +
</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 is not degraded by AmiE, but 3OC12HSL is degraded by AmiE.</p>
 
<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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<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</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-luxR-tt</span>-Plux-<span style="font-style:italic;">rbs-gfp</span> (pSB6A1)</p></li>
 
<li><p class="normal_text">Ptet-<span style="font-style:italic;">rbs-luxR-tt</span>-Plux-<span style="font-style:italic;">rbs-gfp</span> (pSB6A1)</p></li>
 +
<div align="center"><img src="https://static.igem.org/mediawiki/2016/3/35/T--Tokyo_tech--Fig.4_1_5.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</span>
 
</ul>
 
</ul>
 
</div><!-- /best_basic_part_contents -->
 
</div><!-- /best_basic_part_contents -->

Revision as of 13:54, 18 October 2016

Parts Overview

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 did not work, because Prhl activity was too weak compared to Plux. (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, but we noticed that they were inappropriate for two reasons (see the this page). Then, we decided to improve Prhl Promoter and obtain our original improved Prhl (Noticeable Mutant) (BBa_K1949060), hereafter referred to as Prhl(NM), that suited our goal.

Our purpose is to create strong Prhl for our final genetic circuits.

This experiment consists of the three parts below.

  1. Improved Prhl by iGEM 2014 Tokyo_Tech and characterization of rhlR.


    2. Fig. 4-1-1

  2. Improvement of the wild type Prhl.


    3. Fig. 4-1-2

  3. Comparison of the improved Prhl by iGEM 2014 Tokyo_Tech to our original improved Prhl


    4. Fig. 4-1-3

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, see this !

2.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.


Fig. 4-1-4

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-rbs-amiE(pSB6A1)

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

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


    • Fig. 4-1-5

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.