Difference between revisions of "Team:Jilin China/Improve"

 
(6 intermediate revisions by 3 users not shown)
Line 8: Line 8:
 
     <div class="h1_title">1.Improvement in detecting method in DmpR response intensity.</div>
 
     <div class="h1_title">1.Improvement in detecting method in DmpR response intensity.</div>
 
<div class="h1_content">
 
<div class="h1_content">
−
<p>(1) We realize the quantitative measurement in DmpR response intensity using Renilla Luciferase. Compared to GFP <a href="https://2013.igem.org/Team:Peking/Project/BioSensors/DmpR">team Peking-2013</a> used we can detect DmpR response intensity more accurately.
+
<p>(1) We realized the quantitative measurement in DmpR response intensity using Renilla Luciferase. Compared to GFP <a href="https://2013.igem.org/Team:Peking/Project/BioSensors/DmpR">team Peking-2013</a> used, we can detect DmpR response intensity more accurately.
 
<div class="pic_box center">
 
<div class="pic_box center">
 
    <img src="https://static.igem.org/mediawiki/2017/4/4f/T--Jilin_China--improvement01.png"width="500"  /><br />
 
    <img src="https://static.igem.org/mediawiki/2017/4/4f/T--Jilin_China--improvement01.png"width="500"  /><br />
Line 15: Line 15:
 
    </div>
 
    </div>
 
</p>
 
</p>
−
<p>(2) Renilla Luciferase enables us to conduct the measurement in vivo, which provides an easier and faster method in detecting.</p>
+
<p>(2) Renilla Luciferase enabled us to conduct the measurement <i>in vivo</i>, which provided an easier and faster method in detecting.</p>
 
</div>
 
</div>
 
 
 
<div class="h1_title">2.Improvement in response intensity toward certain phenolics.</div>
 
<div class="h1_title">2.Improvement in response intensity toward certain phenolics.</div>
 
<div class="h1_content">
 
<div class="h1_content">
−
<p>Team Jilin_China in 2017 do several mutagenesis in sensor domain of DmpR. To compare the difference between <a href="https://2013.igem.org/Team:Peking/Project/BioSensors/DmpR">Peking-2013 DmpR</a> and our mutated DmpR downstreaming of the same constitutive promoter, we chose phenol, 2-CP, 4-CP, 2,4-DCP and pyrocatechol as the inducer. Expression level of Renilla Luciferase downstream of <i>dmp</i> operon (P<sub>0</sub> promoter) was used to indicate the response intensity towards different phenolics. Comparisons are based on ratio of different DmpR response and mock's toward phenol, 2-CP, 4-CP, 2,4-DCP and pyrocatechol respectively.
+
<p>Team Jilin_China in 2017 did several mutagenesis in sensor domain of DmpR. To compare the difference between <a href="https://2013.igem.org/Team:Peking/Project/BioSensors/DmpR">Peking-2013 DmpR</a> and our mutated DmpR downstreaming of the same constitutive promoter, we chose phenol, 2-CP, 4-CP, 2,4-DCP and pyrocatechol as the inducer. Expression level of Renilla Luciferase downstream of <i>dmp</i> operon (P<sub>0</sub> promoter) was used to indicate the response intensity towards different phenolics. Comparisons were based on ratio of different DmpR response and mock's toward phenol, 2-CP, 4-CP, 2,4-DCP and pyrocatechol respectively.
 
<div class="pic_box center">
 
<div class="pic_box center">
 
    <img src="https://static.igem.org/mediawiki/2017/b/be/T--Jilin_China--improvement03.png"width="500"  /><br />
 
    <img src="https://static.igem.org/mediawiki/2017/b/be/T--Jilin_China--improvement03.png"width="500"  /><br />
Line 29: Line 29:
 
It turned out that both DmpR can response to phenol, 2-CP, pyrocatechol. However, compared to Peking-DmpR, our DmR showed better response to phenol and pyrocatechol.
 
It turned out that both DmpR can response to phenol, 2-CP, pyrocatechol. However, compared to Peking-DmpR, our DmR showed better response to phenol and pyrocatechol.
 
<div class="pic_box center">
 
<div class="pic_box center">
−
    <img src="https://static.igem.org/mediawiki/2017/7/7c/T--Jilin_China--improvement04.png" /><br />
+
    <img src="https://static.igem.org/mediawiki/2017/7/7c/T--Jilin_China--improvement04.png" width="60%" /><br />                              
−
    Figure 3. DmpR response sensitivity toward different phenolics. Jilin-2017-DmpR showed better response to phenol and pyrocatechol.
+
    Figure 3. DmpR response sensitivity toward different phenolics. Jilin-2017-DmpR showed better response to phenol and pyrocatechol.&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;★，p<0.05
−
    </div>
+
            </div>
 
</p>
 
</p>
 
</div>
 
</div>

Latest revision as of 01:27, 2 November 2017

1.Improvement in detecting method in DmpR response intensity.

(1) We realized the quantitative measurement in DmpR response intensity using Renilla Luciferase. Compared to GFP team Peking-2013 used, we can detect DmpR response intensity more accurately.



Figure 1. DmpR circuit. (A) Peking-2013 (B) Jilin-2017

(2) Renilla Luciferase enabled us to conduct the measurement in vivo, which provided an easier and faster method in detecting.

2.Improvement in response intensity toward certain phenolics.

Team Jilin_China in 2017 did several mutagenesis in sensor domain of DmpR. To compare the difference between Peking-2013 DmpR and our mutated DmpR downstreaming of the same constitutive promoter, we chose phenol, 2-CP, 4-CP, 2,4-DCP and pyrocatechol as the inducer. Expression level of Renilla Luciferase downstream of dmp operon (P0 promoter) was used to indicate the response intensity towards different phenolics. Comparisons were based on ratio of different DmpR response and mock's toward phenol, 2-CP, 4-CP, 2,4-DCP and pyrocatechol respectively.


Figure 2. The device we constructed to do comparison between peking-2013 dmpR and our mutated DmpR


It turned out that both DmpR can response to phenol, 2-CP, pyrocatechol. However, compared to Peking-DmpR, our DmR showed better response to phenol and pyrocatechol.


Figure 3. DmpR response sensitivity toward different phenolics. Jilin-2017-DmpR showed better response to phenol and pyrocatechol.       ★，p<0.05