Team:Hannover/Project

From 2014.igem.org

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<h1>Old routes – new ideas</h1>
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<h3>Old routes – new ideas</h3>
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<p class="text">Since 2007, 17 teams from 8 different countries have already chosen the problem of heavy metals in our environment as their topic. A total of 13 medals have been awarded in this subject area. This shows how relevant and highly topical the subject is. While other teams have concentrated more on detection and quantification, we want to go one step further with our concept of plant-based water and soil decontamination.<a href="https://2014.igem.org/Team:Hannover/Background_Project" target="_blank"> Here you find more Background about heavy metals...</a><br><br>With our project we combine cleaning nature by using nature itself: We want to equip plants with a protein which binds four different heavy metals at the same time and hence brings about a significant reduction in the heavy metal concentration of grounds and seas. We thus hope to achieve more extensive binding of hazardous heavy metals than that achieved by conventional methods.</p>
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<p class="text">Since 2007, 17 teams from 8 different countries have already chosen the problem of heavy metals in our environment as their topic. A total of 13 medals have been awarded in this subject area. This shows how relevant and highly topical the subject is. While other teams have concentrated more on detection and quantification, we want to go one step further with our concept of plant-based water and soil decontamination. We want to equip plants with a protein which binds several heavy metals at the same time and hence brings about a significant reduction in the heavy metal concentration. We thus hope to achieve more extensive binding of hazardous heavy metals than that achieved by conventional methods. <a href="https://2014.igem.org/Team:Hannover/Background_Project" target="_blank"> More Background...</a></p>
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<h3>Our path to finding a solution</h3>
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3. step: Quantitative Analysis
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3. step: Water and soil decontamination
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<h1>Our path to finding a solution</h1>
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<p class="text">We created a protein for binding heavy metals, being expressed and produced in plants. This protein, called Top4-Metal-Binding-Protein (T4-MBP), first contains a signal peptide of the expansin-A4-protein (EXP) to secrete the protein to the extracellular space, getting bound there by the cellulose-binding protein A (CBD), which is expressed by the last domain in the T4-MBP. Between those domains there are 4 domains of methallothioneins, each specific for a different heavy metal: copper, arsenic, zinc and cadmium. The transformation of the T4-MBP is done via the binary vector pORE-E3 in which we exchanged the promoter and completed with an optimized 5'UTR.<a href="" target="_blank"> Here you find more about our construct...</a></p>
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<p class="text">Our goal is to produce a Top4 Metal Binding Protein (T4-MBP) which attaches itself to the cellulose of the plants and binds four heavy metals of global relevance. We intend to use naturally occurring metallothioneins - proteins whose specific amino acid sequences alone make them able to form complexes with heavy metals. We have decided on the following heavy metals or domains and combined them to our first synthesis construct on the basis of the cDNA sequences:</p>
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<h1>Our subprojects</h1>
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<tr><td style="border:0px"><h4>Subproject</h4></td><td><h4>How?</h4></td></tr>
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<p class="text">The aim is to be able to use the protein in terrestrial (<i>Arabidopsis thaliana</i>) as well as aquatic plants (Wolffia). Since no-one has had much experience with the transformation of Wolffia, we decided to demonstrate the principle initially in the model organisms <i>A. thaliana</i> and <i>Nicotiana tabacum</i>. Our genetic construct is ultimately to be brought into the target organism with the aid of a transformation by <i>Rhizobium radiobacter</i> (formerly <i>Agrobacterium tumefaciens</i>).</p>
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<tr><td style="border:0px">Constuction of our T4MBP sequenz</td><td>Selection, research and bioinformatics</i><a href="" target="_blank">...</a></td></tr>
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<tr><td style="border:0px">T4MBP Plant</td><td>Stabile transformation of <i>Arabidopsis thaliana</i><a href="" target="_blank">...</a></td></tr>
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<tr><td style="border:0px">Quantitative Analysis of our T4MBP</td><td>Using <i>E. coli</i> as an expression system <a href="" target="_blank">...</a></td></tr>
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<tr><td style="border:0px">Quantitative Analysis of our T4MBP</td><td>Using <i>Nicotiana tabacum</i> as an transient expression system</td></tr>
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<tr><td style="border:0px">Locate the CBD precisely within a plant cell</td><td>CBD fused with GfP, insert it transiently via leaf-infiltration and take a look using a confocal microscope</td></tr>
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<p id="Fusszeile">Fusszeile</p>
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Revision as of 12:45, 7 October 2014

Old routes – new ideas

Since 2007, 17 teams from 8 different countries have already chosen the problem of heavy metals in our environment as their topic. A total of 13 medals have been awarded in this subject area. This shows how relevant and highly topical the subject is. While other teams have concentrated more on detection and quantification, we want to go one step further with our concept of plant-based water and soil decontamination. Here you find more Background about heavy metals...

With our project we combine cleaning nature by using nature itself: We want to equip plants with a protein which binds four different heavy metals at the same time and hence brings about a significant reduction in the heavy metal concentration of grounds and seas. We thus hope to achieve more extensive binding of hazardous heavy metals than that achieved by conventional methods.

1. step: Transformation of Plants 2. step: Secretion and Immobilising 3. step: Water and soil decontamination

Our path to finding a solution

We created a protein for binding heavy metals, being expressed and produced in plants. This protein, called Top4-Metal-Binding-Protein (T4-MBP), first contains a signal peptide of the expansin-A4-protein (EXP) to secrete the protein to the extracellular space, getting bound there by the cellulose-binding protein A (CBD), which is expressed by the last domain in the T4-MBP. Between those domains there are 4 domains of methallothioneins, each specific for a different heavy metal: copper, arsenic, zinc and cadmium. The transformation of the T4-MBP is done via the binary vector pORE-E3 in which we exchanged the promoter and completed with an optimized 5'UTR. Here you find more about our construct...

Our subprojects

Subproject

How?

Constuction of our T4MBP sequenzSelection, research and bioinformatics...
T4MBP PlantStabile transformation of Arabidopsis thaliana...
Quantitative Analysis of our T4MBPUsing E. coli as an expression system ...
Quantitative Analysis of our T4MBPUsing Nicotiana tabacum as an transient expression system
Locate the CBD precisely within a plant cellCBD fused with GfP, insert it transiently via leaf-infiltration and take a look using a confocal microscope

Fusszeile

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