Difference between revisions of "Team:WashU StLouis/Results"

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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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<h5>What should this page contain?</h5>
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<p style="font-size:4vw; text-align:center">Results</p>
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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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<h5>You should also describe what your results mean: </h5>
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    <p style="font-size: 2.5vw; text-align:center; padding 0.5vw">Dsup</p>
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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/2017/b/ba/Results01.png" style="width:65vw; float:center; margin:2vw"/>
  
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    <p style="font-size:1.5vw">The cells that were transformed to contain Dsup were found to have significantly higher survival rates than those without Dsup. Overall, cells containing Dsup--both induced and uninduced--showed an average survival rate of approximately 26% after four hours of exposure to UV-B radiation, while the cells containing only blue chromoprotein (without any radiation resistance genes) showed a survival rate of 0% after four hours. The most pronounced difference between cells containing Dsup and cells containing no radiation resistance was found at three hours, with Dsup-containing cells having an average survival rate of 49% and non-Dsup-containing cells having an average survival rate of 2.3%, meaning that cells with Dsup survived at a rate that was, on average, 2130% higher than the rate at which non-Dsup cells survived.
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    <p style="font-size:1.5vw">There was no significant difference observed in the survival rates of cells in which expression of Dsup was induced via addition of IPTG and cells in which it was not induced, indicating extremely leaky expression in the Lac promoter. </p>
  
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<h5> Project Achievements </h5>
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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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<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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    <p style="font-size: 2.5vw; text-align:center; padding 0.5vw">Failed Expression and mRNA Secondary Structures</p>
  
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<p style="font-size:1.5vw">Although the constructs were successfully transformed into E. coli and showed expression of blue chromoprotein, our reporter, we were not able to obtain any conclusive data on the radiation resistance capabilities of phrAc, phrAt, and uvsE. </p>
  
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<h5>Inspiration</h5>
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<img src="https://static.igem.org/mediawiki/2017/f/f0/Results5.jpeg" style="width:28vw; float:center; margin:2vw"/>
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<p>See how other teams presented their results.</p>
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<p style="font-size:1.5vw">A protein expression test was performed on uvsE + blue chromoprotein (BCP), phrAc + BCP, and phrAt + BCP in order to determine the cause of the failures of our trials. The expression test showed expression of BCP in the constructs containing uvsE and phrAc but no expression of the radiation resistance genes themselves, indicating that it is likely that the mRNA was successfully transcribed but problems occurred during translation of the radiation resistance genes. In the construct containing phrAt, there did appear to be expression of the phrAt gene, but no radiation resistance was observed. </p>
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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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<img src="https://static.igem.org/mediawiki/2017/4/40/Results03.png" style="width:65vw; float:center; margin:2vw"/>
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</ul>
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    <p style="font-size:1.5vw">The failure of uvsE, phrAc, and phrAt to provide any radiation resistance can likely be attributed to the strong secondary structures present in the radiation resistance portions of these constructs and minimal secondary structures present in the BCP portion of the constructs. This is consistent with the observation that blue colonies were present on the plates on which these cells were grown, but no radiation resistance was observed.</p>
  
 
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Latest revision as of 23:06, 1 November 2017

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Results

Dsup

The cells that were transformed to contain Dsup were found to have significantly higher survival rates than those without Dsup. Overall, cells containing Dsup--both induced and uninduced--showed an average survival rate of approximately 26% after four hours of exposure to UV-B radiation, while the cells containing only blue chromoprotein (without any radiation resistance genes) showed a survival rate of 0% after four hours. The most pronounced difference between cells containing Dsup and cells containing no radiation resistance was found at three hours, with Dsup-containing cells having an average survival rate of 49% and non-Dsup-containing cells having an average survival rate of 2.3%, meaning that cells with Dsup survived at a rate that was, on average, 2130% higher than the rate at which non-Dsup cells survived.

There was no significant difference observed in the survival rates of cells in which expression of Dsup was induced via addition of IPTG and cells in which it was not induced, indicating extremely leaky expression in the Lac promoter.

Failed Expression and mRNA Secondary Structures

Although the constructs were successfully transformed into E. coli and showed expression of blue chromoprotein, our reporter, we were not able to obtain any conclusive data on the radiation resistance capabilities of phrAc, phrAt, and uvsE.

A protein expression test was performed on uvsE + blue chromoprotein (BCP), phrAc + BCP, and phrAt + BCP in order to determine the cause of the failures of our trials. The expression test showed expression of BCP in the constructs containing uvsE and phrAc but no expression of the radiation resistance genes themselves, indicating that it is likely that the mRNA was successfully transcribed but problems occurred during translation of the radiation resistance genes. In the construct containing phrAt, there did appear to be expression of the phrAt gene, but no radiation resistance was observed.

The failure of uvsE, phrAc, and phrAt to provide any radiation resistance can likely be attributed to the strong secondary structures present in the radiation resistance portions of these constructs and minimal secondary structures present in the BCP portion of the constructs. This is consistent with the observation that blue colonies were present on the plates on which these cells were grown, but no radiation resistance was observed.