Team:TU Delft-Leiden/WetLab/landmine

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        Module Landmine Detection
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<h2> Landmine Module</h2>
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<h3> Background Information</h3>
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        Information with respect to literature consulted regarding the Modules is referred to under Context. Also, each of the three complementary Modules is equipped with an Integration of Departments, in which it is described how the Departments Modeling, Experimental Work and Microfluidics interact. Furthermore, each Module contains information on Cloning and results are presented under Characterization.
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On the literature [1] a very interesting type of biosensor was found: two natural promoters of Escherichia coli (ybiJ and yqjF) were found to be activated in the presence of some aromatic N-based compounds such as 2,4,6-Trinitrotoluene (2,4,6-TNT), 2,4-Dinitrotoluene (2,4-DNT) and 1,3-Dinitrobenzene (1,3-DNB). Land mines are mainly composed of 2,4,6-TNT, but many times impurities of 2,4-DNT and 1,3-DNB are also present. These last two compounds are more volatile than 2,4,6-TNT and, therefore, they can more easily leak out of the land mine. As a consequence, Belkin and co-workers envisioned in the two aforementioned promoters (ybiJ and yqjF) a high potential to develop a biosensor for land mine detection.
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This was a very attractive case of study to implement the Plug-and-Play biosensor based on electrical current developed by our team.
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        <ul>Module Landmine Detection
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            <li><a href="/Team:TU_Delft-Leiden/WetLab/landmine/theory">Context</a></li>
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            <li><a href="/Team:TU_Delft-Leiden/Project/Life_science/landmine/integration">Integration of Departments</a></li>
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            <li><a href="/Team:TU_Delft-Leiden/WetLab/landmine/cloning">Cloning</a></li>
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            <li><a href="/Team:TU_Delft-Leiden/WetLab/landmine/characterisation">Characterization</a></li>
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Keen to see our <b>conclusions</b> for this module?  See the list below!
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<h3> Coupling the promoters to our system </h3>
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As a final goal, it was desired to link the ybiJ and yqjF promoters to the generation of an electrical current. Therefore, it was our intention to express two of the three genes of the mtrCAB pathway independently (either constitutively or regulated via an inducer) and the last mtrCAB gene regulated by one of the two promoters induced by land mine compounds. Hence, the presence of land mines would trigger the expression of the lacking gene of the mtrCAB pathway, and therefore, it would start the secreation of electrons out of the cell. This way, it was intended to correlate the presence of the abovementioned chemicals present in land mines with a certain electrical current.
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<p> It was, nevertheless, more convenient to test the selected promoters with a more easily measurable reporting system
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<li>The two best BioBricks for Landmine detection are LD5 (p[J] incl. N-enzymes) and LD4 (p[F] incl. N-enzymes). The N-genes contribute to sensitivity of the promoters.</li>  
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<li> The LD5 construct seems to be the most suitable construct for Landmine Detection. </li>
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<li> Evidence suggests that the ybiJ and yqjF promoters are activated by 2,4-DNT, but it cannot be totally discarded the possibility that acetonitrile is the trigger of the response of these promoters. </li></ul></p>
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<h3> Choosing a reporter gene </h3>
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<p>Want to know how we came to these conclusions? Go to our <a href= https://2014.igem.org/Team:TU_Delft-Leiden/WetLab/landmine/characterisation><b>Characterization</b></a> page!
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To initially check the functionality of the ybiJ and yqjF promoters, a fluorescent protein mKate2 was chosen as a reporter. mKate2 was selected instead of the conventional Green Fluorescent Protein (GFP) because it has been observed to be more evenly expressed among the different cells of a cellular culture than GFP.
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        Interested in one of our other Modules? Navigate to the <a href=" https://2014.igem.org/Team:TU_Delft-Leiden/Project/Life_science/EET"> <b> Module Electron Transport </b> </a> where you can find everything regarding the implementation of the MtrCAB conduit. Interested in living materials? Go to our <a href="https://2014.igem.org/Team:TU_Delft-Leiden/Project/Life_science/curli"> <b> Module Conductive Curli </b> </a> project and find out how you can combine the benefits of both living- and non-living materials.
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<h2> Cloning Strategy and Characterisation of this module</h2>
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  <li><a href="/Team:TU_Delft-Leiden/WetLab/landmine/cloning"><p>Cloning</p>
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        <img title="Deminer" src="https://static.igem.org/mediawiki/2014/5/5b/Deminer.jpg" style="display:block;margin-left:auto;margin-right:auto;height:50%;width:50%;" />
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  <li><a href="/Team:TU_Delft-Leiden/WetLab/landmine/characterisation"><p>Characterisation</p>
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<h3> References </h3>
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<p>[1] S. Yagur-Kroll, S. Belkin <i>et al.</i>, “<i>Escherichia Coli</i> bioreporters for the detection of 2,4-dinitrotoluene and 2,4,6-trinitrotoluene”, Appl. Microbiol. Biotechnol. 98, 885-895, 2014. </p>
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Latest revision as of 20:54, 17 October 2014

Module Landmine Detection

Information with respect to literature consulted regarding the Modules is referred to under Context. Also, each of the three complementary Modules is equipped with an Integration of Departments, in which it is described how the Departments Modeling, Experimental Work and Microfluidics interact. Furthermore, each Module contains information on Cloning and results are presented under Characterization.

Keen to see our conclusions for this module? See the list below!

  • The two best BioBricks for Landmine detection are LD5 (p[J] incl. N-enzymes) and LD4 (p[F] incl. N-enzymes). The N-genes contribute to sensitivity of the promoters.
  • The LD5 construct seems to be the most suitable construct for Landmine Detection.
  • Evidence suggests that the ybiJ and yqjF promoters are activated by 2,4-DNT, but it cannot be totally discarded the possibility that acetonitrile is the trigger of the response of these promoters.

Want to know how we came to these conclusions? Go to our Characterization page!


Interested in one of our other Modules? Navigate to the Module Electron Transport where you can find everything regarding the implementation of the MtrCAB conduit. Interested in living materials? Go to our Module Conductive Curli project and find out how you can combine the benefits of both living- and non-living materials.

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