Difference between revisions of "Team:ITB Indonesia/Demonstrate"

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<h3>★  ALERT! </h3>
 
<p>This page is used by the judges to evaluate your team for the <a href="https://2017.igem.org/Judging/Medals">medal criterion</a> or <a href="https://2017.igem.org/Judging/Awards"> award listed above</a>. </p>
 
<p> Delete this box in order to be evaluated for this medal criterion and/or award. See more information at <a href="https://2017.igem.org/Judging/Pages_for_Awards"> Instructions for Pages for awards</a>.</p>
 
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<h1 class="ITB_h1">Demonstrate</h1>
  
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<h1>Demonstrate</h1>
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<h3>Gold Medal Criterion #4</h3>
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Teams that can show their system working under real world conditions are usually good at impressing the judges in iGEM. To achieve gold medal criterion #4, convince the judges that your project works. There are many ways in which your project working could be demonstrated, so there is more than one way to meet this requirement. This gold medal criterion was introduced in 2016, so check our what 2016 teams did to achieve a their gold medals!
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<h1 class="ITB_h1" style="padding-bottom: 30px; margin-bottom: 50px; border-bottom: 2px solid #1c2922 !important; padding-left: 30px; text-align: center; color: #1c2922">Demonstrate</h1>
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Please see the <a href="https://2017.igem.org/Judging/Medals">2017 Medals Page</a> for more information.
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<h4> What should we do for our demonstration?</h4>
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<h5> Standard teams </h5>
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If you have built a proof of concept system, you can demonstrate it working under real world conditions. If you have built a biological device that is intended to be a sensor, can you show it detecting whatever it is intended to sense. If it is intended to work in the field, you can show how this might work using a simulated version in the lab, or a simulation of your device in the field.<strong> Please note biological materials must not be taken out of the lab</strong>.
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<p style="font-size: 18px"><strong>Real World's Conditions, Real World's Materials</strong></p>
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<p>In order to demonstrate the work of our project in real world's conditions, we did not only use chemical test reagents (such as pNPB) to test our PETase activity. We instead tried to find the realest material that would be encountered by our bacteria on its applications. We ended up using plastic bottles directly as materials to be degraded.</p>
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<p>The results of commonly found plastic bottle degradation by our bacteria are quite satisfactory. SEM analyses showed that our bacteria are able to degrade even real life plastic bottles, which are not pure PET. The effect is shown by SEM analyses where the plastic surface inoculated by our PETase-containing bacteria are rougher and has more cracks compared to its control.</p>
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<center><img src="https://static.igem.org/mediawiki/parts/a/a0/T--ITB_Indonesia--SEMControl.jpeg" style="width: 500px; height: auto; img-align:center"></center>
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<center><em> Figure 2. SEM result of plastic fragment incubated with control BL21 cells bearing no plasmid </em></center>
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<center><img src="https://static.igem.org/mediawiki/parts/7/72/T--ITB_Indonesia--SEMPETase.jpeg" style="width: 500px; height: auto; img-align:center"></center>
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<center><em> Figure 3. SEM result of plastic fragment incubated with BL21 cells transformed with BBa_K2378005 shows rougher surface with more cracks </em></center>
 
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<h5> Special track teams </h5>
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<center><img src="https://static.igem.org/mediawiki/parts/c/c2/T--ITB_Indonesia--SEMNhaR.jpeg" style="width: 500px; height: auto; img-align:center"></center>
 
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<center><em> Figure 4. SEM result of plastic fragment incubated with BL21 cells transformed with BBa_K2378006 shows rougher surface with more cracks </em></center>
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Special track teams can achieve this medal criterion by bringing their work to the Jamboree and showcasing it in the track event. Art & Design, Measurement, Hardware and Software tracks will all have showcase events at the Giant Jamboree.<strong> Please note biological materials must not be taken out of the lab</strong>.
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<p style="font-size: 18px"><strong>Real World's Conditions, Real World's Inducers</strong></p>
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<p>Aside from using daily materials, we also tried to incorporate the use of real world's inducers. Our bacteria are supposed to be induced naturally when they encounter plastic, due to the presence of organic pollutants (OPs) on the surface of the plastic.</p>
 
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<p>In order to mimic real world's conditions, we also tried to induce the expression of PETase in our bacteria through exposure with plastic wastes.</p>
 
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<p>The culture induced by plastic wastes did not have the same expression rate as cultures induced with common chemical inducer salicylate. However, it turned out that some degree of expression is observed in cultures induced with plastic wastes, albeit the difference of the expression rate compared to uninduced cultures are not too significant.</p>
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<center><img src="https://static.igem.org/mediawiki/parts/6/6b/T--ITB_Indonesia--psalpnpb.jpeg" style="width: 500px; height: auto; img-align:center"></center>
 
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<center><em> Figure 5. pSal-Regulated PETase Activity (pNPB Assay) of BBa K2378004 Transformants</em></center>
 
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Latest revision as of 03:26, 2 November 2017


Demonstrate


Demonstrate

Real World's Conditions, Real World's Materials

In order to demonstrate the work of our project in real world's conditions, we did not only use chemical test reagents (such as pNPB) to test our PETase activity. We instead tried to find the realest material that would be encountered by our bacteria on its applications. We ended up using plastic bottles directly as materials to be degraded.

The results of commonly found plastic bottle degradation by our bacteria are quite satisfactory. SEM analyses showed that our bacteria are able to degrade even real life plastic bottles, which are not pure PET. The effect is shown by SEM analyses where the plastic surface inoculated by our PETase-containing bacteria are rougher and has more cracks compared to its control.

Figure 2. SEM result of plastic fragment incubated with control BL21 cells bearing no plasmid

Figure 3. SEM result of plastic fragment incubated with BL21 cells transformed with BBa_K2378005 shows rougher surface with more cracks

Figure 4. SEM result of plastic fragment incubated with BL21 cells transformed with BBa_K2378006 shows rougher surface with more cracks

Real World's Conditions, Real World's Inducers

Aside from using daily materials, we also tried to incorporate the use of real world's inducers. Our bacteria are supposed to be induced naturally when they encounter plastic, due to the presence of organic pollutants (OPs) on the surface of the plastic.

In order to mimic real world's conditions, we also tried to induce the expression of PETase in our bacteria through exposure with plastic wastes.

The culture induced by plastic wastes did not have the same expression rate as cultures induced with common chemical inducer salicylate. However, it turned out that some degree of expression is observed in cultures induced with plastic wastes, albeit the difference of the expression rate compared to uninduced cultures are not too significant.

Figure 5. pSal-Regulated PETase Activity (pNPB Assay) of BBa K2378004 Transformants