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

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<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 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.
 
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    Figure 1. DmpR circuit. (A) Peking-2013 (B) Jilin-2017
 
    Figure 1. DmpR circuit. (A) Peking-2013 (B) Jilin-2017
 
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<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 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.
 
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    <img src="https://static.igem.org/mediawiki/2017/b/be/T--Jilin_China--improvement03.png"  /><br />
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    <img src="https://static.igem.org/mediawiki/2017/b/be/T--Jilin_China--improvement03.png"width="500"  /><br />
 
    Figure 1. The device we constructed to do comparison between peking-2013 dmpR and our mutated DmpR
 
    Figure 1. The device we constructed to do comparison between peking-2013 dmpR and our mutated DmpR
 
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Revision as of 19:14, 31 October 2017

1.Improvement in detecting method in DmpR response intensity.

(1) We realize 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 enables us to conduct the measurement in vivo, which provides an easier and faster method in detecting.

2.Improvement in response intensity toward certain phenolics.

Team Jilin_China in 2017 do 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 are based on ratio of different DmpR response and mock's toward phenol, 2-CP, 4-CP, 2,4-DCP and pyrocatechol respectively.


Figure 1. 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 2. DmpR response sensitivity toward different phenolics. Jilin-2017-DmpR showed better response to phenol and pyrocatechol.