Difference between revisions of "Team:TUST China/Results"

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        function backtop(){
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<h1>Results</h1>
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<p>Here you can describe the results of your project and your future plans. </p>
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<h3>What should this page contain?</h3>
 
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<ul>
 
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<li> Clearly and objectively describe the results of your work.</li>
 
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<li> Future plans for the project. </li>
 
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<li> Considerations for replicating the experiments. </li>
 
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</ul>
 
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</div>
 
  
  
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<div class="column two_thirds_size" >
 
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<h3>Describe what your results mean </h3>
 
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<ul>
 
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<li> Interpretation of the results obtained during your project. Don't just show a plot/figure/graph/other, tell us what you think the data means. This is an important part of your project that the judges will look for. </li>
 
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<li> Show data, but remember all measurement and characterization data must be on part pages in the Registry. </li>
 
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<li> Consider including an analysis summary section to discuss what your results mean. Judges like to read what you think your data means, beyond all the data you have acquired during your project. </li>
 
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    <img src="https://static.igem.org/mediawiki/2018/b/b5/T--TUST_China--result.png"  style="width: 30%;margin-top:50px;"/>
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<h3> Project Achievements </h3>
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<p>You can also include a list of bullet points (and links) of the successes and failures you have had over your summer. It is a quick reference page for the judges to see what you achieved during your summer.</p>
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<li>A list of linked bullet points of the successful results during your project</li>
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    <div class="content-header" style="margin-top: 0px;">
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<li>A list of linked bullet points of the unsuccessful results during your project. This is about being scientifically honest. If you worked on an area for a long time with no success, tell us so we know where you put your effort.</li>
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        <h1 style="top: 160px;text-align: center;font-size: 3.4vw;margin:10px auto;">Result
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        </h1>
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        <div class="head-content"> <p style="color: #fff;text-align: center">After building the chassis strain, we began to design experiments to verify whether it could achieve our desired results.</p>
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<h3>Inspiration</h3>
 
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<p>See how other teams presented their results.</p>
 
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<ul>
 
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<li><a href="https://2014.igem.org/Team:TU_Darmstadt/Results/Pathway">2014 TU Darmstadt </a></li>
 
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<li><a href="https://2014.igem.org/Team:Imperial/Results">2014 Imperial </a></li>
 
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<li><a href="https://2014.igem.org/Team:Paris_Bettencourt/Results">2014 Paris Bettencourt </a></li>
 
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</ul>
 
 
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        <h2>For the Detecting chasis:</h2>
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        <p>Since our chassis strain is E. coli and does not contain tetracycline resistance, we need to verify whether the gene tet(A), tet(R), tet(O) we introduced enables E. coli to survive in water samples containing tetracycline and achieve detection purposes.
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            Firstly, we need to test the tetracycline tolerance of the constructive detection chasis, here is the resluts in the fig.1
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        </p>
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        <img src="https://static.igem.org/mediawiki/2018/e/e9/T--TUST_China--RESULT1.png" width="80%">
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        <center><span>fig.1 The tetracycline tolerance of detecting parts comparing with the orginal E.coli</span></center>
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        <p>From the data, we can clearly see that, the constructive detection chasis obtain a tetracycline tolerance from 0-100μg/μL, and then, detecting chasis can be growth normally in the culture medium with tetracycline.</p>
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        <p>Meanwhile, we have a collaboration with the NKU-China that we sent our detecting chasis to ask for measuring minimal inhibitory concentration (MIC), and we can also get the tetracycline tolerance of this chasis which similar with our test results. </p>
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        <img src="https://static.igem.org/mediawiki/2018/5/5f/T--TUST_China--RESULT2.png" width="80%">
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        <center><span>fig.2 The MIC of the detecting parts</span></center>
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        <p>After determining that our chassis can survive tetracycline, we need to further test its performance and get the following data:</p>
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        <img src="https://static.igem.org/mediawiki/2018/1/12/T--TUST_China--RESULT3.png" width="80%">
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        <center><span>fig.3 The ratio of FL value and ABS value of the detection chasis under different tc concentrations</span></center>
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        <p>In order to further improve the accuracy of detection, we have exchanged the gene tet(A) and gene gfp. we verified those constructive chasis with the same
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            experiment designs, and then we found that it can detect more lower concentration at 0.00005μg/ml.</p>
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        <img src="https://static.igem.org/mediawiki/2018/e/e3/T--TUST_China--RESULT4.png" width="80%">
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        <center><span>fig.4 The ABS value of the detection chasis gene exchange under different tc concentrations</span></center>
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        <p>The detecting chasis with gene exchanged can detect the lower concentration of tetracycline at 0.00005μg/ml in the water samples, but it need more time to growth and this section
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            is not a stable chasis for each time to detect tetracycline.</p>
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        <img src="https://static.igem.org/mediawiki/2018/8/87/T--TUST_China--RESULT5.png" width="80%">
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        <center><span>fig.5 The ABS value of the detection chasis gene exchange under different tc concentrations</span></center>
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        <p>Considering the stability of the project, we prefer to use the original tetracycline detection chasis to detect water samples.</p>
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        <h2>For the Degradation chasis:</h2>
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        <p>We successfully constructed four degradation chasis with lignin peroxidase, Manganese peroxidase and laccase. After the verification experiments,
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            we got lots of data about those enzymes performances under the co-culture system in the below:</p>
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        <img src="https://static.igem.org/mediawiki/2018/9/9d/T--TUST_China--RESULT6.png" width="80%">
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        <img src="https://static.igem.org/mediawiki/2018/1/18/T--TUST_China--RESULT7.png" width="80%">
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        <img src="https://static.igem.org/mediawiki/2018/a/a8/T--TUST_China--RESULT8.png" width="80%">
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        <img src="https://static.igem.org/mediawiki/2018/a/a8/T--TUST_China--result9.png" width="80%">
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        <p>Through the data comparison above, we can get that the lignin peroxidase perform the better ability to degrade the tetracycline in the culture media and Manganese peroxidase cannot work well alone. So we retain the composite parts that including lignin peroxidase and Manganese peroxidase to degrade tetracycline together in this parts.</p>
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{{TUST_China/foot}}

Latest revision as of 23:59, 17 October 2018

nav

Result

After building the chassis strain, we began to design experiments to verify whether it could achieve our desired results.

For the Detecting chasis:

Since our chassis strain is E. coli and does not contain tetracycline resistance, we need to verify whether the gene tet(A), tet(R), tet(O) we introduced enables E. coli to survive in water samples containing tetracycline and achieve detection purposes. Firstly, we need to test the tetracycline tolerance of the constructive detection chasis, here is the resluts in the fig.1

fig.1 The tetracycline tolerance of detecting parts comparing with the orginal E.coli

From the data, we can clearly see that, the constructive detection chasis obtain a tetracycline tolerance from 0-100μg/μL, and then, detecting chasis can be growth normally in the culture medium with tetracycline.

Meanwhile, we have a collaboration with the NKU-China that we sent our detecting chasis to ask for measuring minimal inhibitory concentration (MIC), and we can also get the tetracycline tolerance of this chasis which similar with our test results.

fig.2 The MIC of the detecting parts

After determining that our chassis can survive tetracycline, we need to further test its performance and get the following data:

fig.3 The ratio of FL value and ABS value of the detection chasis under different tc concentrations

In order to further improve the accuracy of detection, we have exchanged the gene tet(A) and gene gfp. we verified those constructive chasis with the same experiment designs, and then we found that it can detect more lower concentration at 0.00005μg/ml.

fig.4 The ABS value of the detection chasis gene exchange under different tc concentrations

The detecting chasis with gene exchanged can detect the lower concentration of tetracycline at 0.00005μg/ml in the water samples, but it need more time to growth and this section is not a stable chasis for each time to detect tetracycline.

fig.5 The ABS value of the detection chasis gene exchange under different tc concentrations

Considering the stability of the project, we prefer to use the original tetracycline detection chasis to detect water samples.

For the Degradation chasis:

We successfully constructed four degradation chasis with lignin peroxidase, Manganese peroxidase and laccase. After the verification experiments, we got lots of data about those enzymes performances under the co-culture system in the below:

Through the data comparison above, we can get that the lignin peroxidase perform the better ability to degrade the tetracycline in the culture media and Manganese peroxidase cannot work well alone. So we retain the composite parts that including lignin peroxidase and Manganese peroxidase to degrade tetracycline together in this parts.

foot