Difference between revisions of "Team:Goettingen/Measurement"

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          <h2>Measurement</h2>
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        <p>Glyphosate is a widely used herbicide with unknown risks for the environment and human health. Our goal is to develop a simple and cheap detection system, which allows average consumers to assess whether glyphosate is present in their food products. The colour-based glyphosate detection system relies on monitoring intraspecies competition of strains of the environmental bacterium <i>Bacillus&nbsp;subtilis</i> tolerating different amounts of the weed killer. To obtain <i>B.&nbsp;subtilis</i> strains that could grow with glyphosate, we have evolved the bacteria in the presence of the weed killer. These experiments revealed that <i>B.&nbsp;subtilis</i> rapidly adapts to glyphosate at the genome level by the acquisition of beneficial mutations, rendering the bacteria insensitive to the herbicide. The characterization of the mutants led to the discovery of the first glyphosate uptake systems! Furthermore, the bacterial strains tolerating different amounts of glyphosate facilitated the construction of our herbicide detection system.
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        <p>Our measurement method uses fluorescence that is linked to competing <i>Bacillus&nbsp;subtilis</i> strains. The sensitive strain is tagged with GFP, while the glyphosate-resistant strain is tagged with mOrange. In a competition assay are both strains 1:1 mixed and grown with a possibly glyphosate-containing sample. The ratio between green and orange fluorescence gives information on the glyphosate concentration. The functionality is shown <a href="https://2018.igem.org/Team:Goettingen/Results#r_Competition" target="_blank">here</a>.
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        <p>Furthermore, we constructed another system that uses the activity of the &beta;&ndash;galactosidase that is expressed in the wildtype-like strain BP193 (<i>trpC2 p<sub>alf4</sub>::lacZ</i>). The glyphosate-resistant strain BP235 (<i>trpC2 amyE::(morange-lacZ-aphA3)</i> &Delta;<i>gltT::spc</i> &Delta;<i>gltP::cat</i> eliminates strain BP193 stronger the more glyphosate is present. This elimination can be monitored <i>via</i> the intensity of the blue color of the medium (<a href="https://2018.igem.org/Team:Goettingen/Results#r_Competition" target="_blank">Results</a>). The intensity is compared to a test strip and the concentration can be read.
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        <p>The functionality of our product is shown in the following picture (Figure 1).</p>
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        <p>Roundup<sup>&reg;</sup> is sprayed on soil, Then, water is applied to solve the glyphosate from the sample. The solution is filtered through a filter devide to avoid any other contaminations like other microorganisms. The sample drops onto solid media powder with all necessary ingredients and <i>Bacillus&nbsp;subtilis</i> spores from the two competing strains. The sample is incubated for 10&nbsp;h at 37°C with agitation. Now, the blue color can be compared to a test strip. The intensity of the blue color determines the concentration of glyphosate in the sample. The usage of our product is also shown in the following movie.
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<h3>★  ALERT! </h3>
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<p>This page is used by the judges to evaluate your team for the <a href="https://2018.igem.org/Judging/Medals">medal criterion</a> or <a href="https://2018.igem.org/Judging/Awards"> award listed below</a>. </p>
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<p> Delete this box in order to be evaluated for this medal criterion and/or award. See more information at <a href="https://2018.igem.org/Judging/Pages_for_Awards"> Instructions for Pages for awards</a>.</p>
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<h1>Measurement</h1>
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<p>There are a lot of exciting parts in the Registry, but many parts have still not been characterized. Synthetic Biology needs great measurement approaches for characterizing new parts, and efficient new methods for characterizing many parts at once. If you've done something exciting in the area of Measurement, describe it here!</p>
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<h3>Best Innovation in Measurement Special Prize</h3>
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<p>If you've done excellent work in measurement, you should consider nominating your team for this special prize. Designing great measurement approaches for characterizing new parts or developing and implementing an efficient new method for characterizing thousands of parts are good examples.
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To compete for the <a href="https://2018.igem.org/Judging/Awards">Best Innovation in Measurement prize</a>, please describe your work on this page and also fill out the description on the <a href="https://2018.igem.org/Judging/Judging_Form">judging form</a>.
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You must also delete the message box on the top of this page to be eligible for this prize.
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<h3>Inspiration</h3>
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<p>You can look at what other teams did to get some inspiration! <br />
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Here are a few examples:</p>
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<li><a href="https://2016.igem.org/Team:Stanford-Brown">2016 Stanford-Brown</a></li>
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<li><a href="https://2016.igem.org/Team:Genspace">2016 Genspace</a></li>
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<li><a href="https://2015.igem.org/Team:William_and_Mary">2015 William and Mary</a></li>
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<li><a href="https://2014.igem.org/Team:Aachen">2014 Aachen  </a></li>
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Revision as of 19:17, 10 October 2018

Measurement

Glyphosate is a widely used herbicide with unknown risks for the environment and human health. Our goal is to develop a simple and cheap detection system, which allows average consumers to assess whether glyphosate is present in their food products. The colour-based glyphosate detection system relies on monitoring intraspecies competition of strains of the environmental bacterium Bacillus subtilis tolerating different amounts of the weed killer. To obtain B. subtilis strains that could grow with glyphosate, we have evolved the bacteria in the presence of the weed killer. These experiments revealed that B. subtilis rapidly adapts to glyphosate at the genome level by the acquisition of beneficial mutations, rendering the bacteria insensitive to the herbicide. The characterization of the mutants led to the discovery of the first glyphosate uptake systems! Furthermore, the bacterial strains tolerating different amounts of glyphosate facilitated the construction of our herbicide detection system.

Our measurement method uses fluorescence that is linked to competing Bacillus subtilis strains. The sensitive strain is tagged with GFP, while the glyphosate-resistant strain is tagged with mOrange. In a competition assay are both strains 1:1 mixed and grown with a possibly glyphosate-containing sample. The ratio between green and orange fluorescence gives information on the glyphosate concentration. The functionality is shown here.

Furthermore, we constructed another system that uses the activity of the β–galactosidase that is expressed in the wildtype-like strain BP193 (trpC2 palf4::lacZ). The glyphosate-resistant strain BP235 (trpC2 amyE::(morange-lacZ-aphA3) ΔgltT::spc ΔgltP::cat eliminates strain BP193 stronger the more glyphosate is present. This elimination can be monitored via the intensity of the blue color of the medium (Results). The intensity is compared to a test strip and the concentration can be read.

The functionality of our product is shown in the following picture (Figure 1).

Roundup® is sprayed on soil, Then, water is applied to solve the glyphosate from the sample. The solution is filtered through a filter devide to avoid any other contaminations like other microorganisms. The sample drops onto solid media powder with all necessary ingredients and Bacillus subtilis spores from the two competing strains. The sample is incubated for 10 h at 37°C with agitation. Now, the blue color can be compared to a test strip. The intensity of the blue color determines the concentration of glyphosate in the sample. The usage of our product is also shown in the following movie.