Difference between revisions of "Team:TU Darmstadt/Project/Kill Switch"
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| − | <p>Since our kill switch is based on linking the availability of an essential gene (product) to the presence of a biofilm, this essential gene first had to be knocked out. In our first approach we chose the gene <i>dnaA</i> for this. Since it encodes for a protein that plays an important role in initiating <b>DNA replication</b> in prokaryotes [1Jillian], its absence would lead to cell death. However, after some time we realized that the choice of this gene caused a problem: <i>dnaA</i> is located in an operon that contains other genes [1Jillian]. In the case of a knockout by destroying the promoter, the entire operon would no longer be transcribed. This could have led to unpredictable complications. | + | <p>Since our kill switch is based on linking the availability of an essential gene (product) to the presence of a biofilm, this essential gene first had to be knocked out. In our first approach we chose the gene <i>dnaA</i> for this. Since it encodes for a protein that plays an important role in initiating <b>DNA replication</b> in prokaryotes |
| + | <sup id="cite_ref-1"><a href="#cite_note-1">[1Jillian]</a></sup> | ||
| + | , its absence would lead to cell death. However, after some time we realized that the choice of this gene caused a problem: <i>dnaA</i> is located in an operon that contains other genes | ||
| + | <sup id="cite_ref-1"><a href="#cite_note-1">[1Jillian]</a></sup> | ||
| + | . In the case of a knockout by destroying the promoter, the entire operon would no longer be transcribed. This could have led to unpredictable complications. | ||
<br> | <br> | ||
<br> | <br> | ||
| − | In addition, dnaA is only essential for cell division, so it is possible that the microorganisms do not die but go into dormancy instead. For this reason, we searched for an easier accessible gene - and found <i>rpsB</i>. Its gene product is the ribosomal protein S2 in the 30S subunit of prokaryotic ribosomes. As one of the biggest ribosomal proteins in the small subunit [2Jillian], RpsB is essential for the functionality of the ribosomes and thus for translation. When a bacterium leaves the biofilm, due to our killswitch RpsB will no longer be produced. This leads to the bacterium being unable to keep up metabolic function, resulting into a fast cell death. Another advantage is that <i>rpsB</i> is the only gene under control of the corresponding operon, therefore being much easier to control.</p> | + | In addition, dnaA is only essential for cell division, so it is possible that the microorganisms do not die but go into dormancy instead. For this reason, we searched for an easier accessible gene - and found <i>rpsB</i>. Its gene product is the ribosomal protein S2 in the 30S subunit of prokaryotic ribosomes. As one of the biggest ribosomal proteins in the small subunit |
| + | <sup id="cite_ref-1"><a href="#cite_note-1">[2Jillian]</a></sup> | ||
| + | , RpsB is essential for the functionality of the ribosomes and thus for translation. When a bacterium leaves the biofilm, due to our killswitch RpsB will no longer be produced. This leads to the bacterium being unable to keep up metabolic function, resulting into a fast cell death. Another advantage is that <i>rpsB</i> is the only gene under control of the corresponding operon, therefore being much easier to control.</p> | ||
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<a class="referencestd" href="https://www.sciencedirect.com/science/article/abs/pii/0167478194900256" | <a class="referencestd" href="https://www.sciencedirect.com/science/article/abs/pii/0167478194900256" | ||
target="_blank">Quelle1Jillian (muss noch ausgeschrieben werden)</a> | target="_blank">Quelle1Jillian (muss noch ausgeschrieben werden)</a> | ||
| + | <a class="anchor" id="cite_note-2"></a> | ||
| + | <a class="referencestd" href="https://link.springer.com/article/10.1007/BF00333661" | ||
| + | target="_blank">Quelle2Jillian (muss noch ausgeschrieben werden)</a> | ||
</div> | </div> | ||
Revision as of 12:47, 13 October 2020
[EDIT] essential gene decision making [EDIT]
Since our kill switch is based on linking the availability of an essential gene (product) to the presence of a biofilm, this essential gene first had to be knocked out. In our first approach we chose the gene dnaA for this. Since it encodes for a protein that plays an important role in initiating DNA replication in prokaryotes
[1Jillian]
, its absence would lead to cell death. However, after some time we realized that the choice of this gene caused a problem: dnaA is located in an operon that contains other genes
[1Jillian]
. In the case of a knockout by destroying the promoter, the entire operon would no longer be transcribed. This could have led to unpredictable complications.
In addition, dnaA is only essential for cell division, so it is possible that the microorganisms do not die but go into dormancy instead. For this reason, we searched for an easier accessible gene - and found rpsB. Its gene product is the ribosomal protein S2 in the 30S subunit of prokaryotic ribosomes. As one of the biggest ribosomal proteins in the small subunit
[2Jillian]
, RpsB is essential for the functionality of the ribosomes and thus for translation. When a bacterium leaves the biofilm, due to our killswitch RpsB will no longer be produced. This leads to the bacterium being unable to keep up metabolic function, resulting into a fast cell death. Another advantage is that rpsB is the only gene under control of the corresponding operon, therefore being much easier to control.
Experts
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At vero eos et
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Former iGEMers
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Lorem ipsum dolor sit amet, consetetur sadipscing elitr, sed diam nonumy eirmod tempor invidunt ut labore et dolore magna aliquyam erat, sed diam voluptua. At vero eos et accusam et justo duo dolores et ea rebum. Stet clita kasd gubergren, no sea takimata sanctus est Lorem ipsum dolor sit amet. Lorem ipsum dolor sit amet, consetetur sadipscing elitr, sed diam nonumy eirmod tempor invidunt ut labore et dolore magna aliquyam erat, sed diam voluptua.
At vero eos et
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Lorem ipsum dolor sit amet, consetetur sadipscing elitr, sed diam nonumy eirmod tempor invidunt ut labore et dolore magna aliquyam erat, sed diam voluptua. At vero eos et accusam et justo duo dolores et ea rebum. Stet clita kasd gubergren, no sea takimata sanctus est Lorem ipsum dolor sit amet. Lorem ipsum dolor sit amet, consetetur sadipscing elitr, sed diam nonumy eirmod tempor invidunt ut labore et dolore magna aliquyam erat, sed diam voluptua. At vero eos et accusam et justo duo dolores et ea rebum. Stet clita kasd gubergren, no sea takimata sanctus est Lorem ipsum dolor sit amet.
Others
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At vero eos et
Lorem ipsum dolor sit amet, consetetur sadipscing elitr, sed diam nonumy eirmod tempor invidunt ut labore et dolore magna aliquyam erat, sed diam voluptua. At vero eos et accusam et justo duo dolores et ea rebum. Stet clita kasd gubergren, no sea takimata sanctus est Lorem ipsum dolor sit amet. Lorem ipsum dolor sit amet, consetetur sadipscing elitr, sed diam nonumy eirmod tempor invidunt ut labore et dolore magna aliquyam erat, sed diam voluptua. At vero eos et
Lorem ipsum dolor sit amet, consetetur sadipscing elitr, sed diam nonumy eirmod tempor invidunt ut labore et dolore magna aliquyam erat, sed diam voluptua. At vero eos et accusam et justo duo dolores et ea rebum. Stet clita kasd gubergren, no sea takimata sanctus est Lorem ipsum dolor sit amet. Lorem ipsum dolor sit amet, consetetur sadipscing elitr, sed diam nonumy eirmod tempor invidunt ut labore et dolore magna aliquyam erat, sed diam voluptua. At vero eos et
Lorem ipsum dolor sit amet, consetetur sadipscing elitr, sed diam nonumy eirmod tempor invidunt ut labore et dolore magna aliquyam erat, sed diam voluptua. At vero eos et accusam et justo duo dolores et ea rebum. Stet clita kasd gubergren, no sea takimata sanctus est Lorem ipsum dolor sit amet. Lorem ipsum dolor sit amet, consetetur sadipscing elitr, sed diam nonumy eirmod tempor invidunt ut labore et dolore magna aliquyam erat, sed diam voluptua. At vero eos et accusam et justo duo dolores et ea rebum. Stet clita kasd gubergren, no sea takimata sanctus est Lorem ipsum dolor sit amet.