Team:Aberdeen Scotland/Parts/Device

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<h1>Background to Parts Design</h1>
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<h1>Detector Test Data</h1>
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<p>Antigen 43 (Ag43), the product of the </i>flu</i> gene, is a cell-surface autotransporter protein found in <i>Escherichia coli</i>. It is expressed at about 50,
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<p>The initial concept of the fluorescence detector device had a very rough design. Thus it was built from bits and pieces in order to do a preliminary test and
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000 copies/cell and is initially synthesised as a precursor of 1039 amino acids. Upon removal of the signal peptide, the protein is transported to the cell surface
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verify its viability.</p>
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and is composed of an α subunit (499 amino acids) at the N-terminus and a β subunit (488 amino acids) at the C-terminus. Ag43 is mainly known to induce cell-to-cell
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aggregation and be involved in biofilm formation. However, as the necessary information required for auto transportation resides in the protein itself, the main of
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<img src="https://static.igem.org/mediawiki/2014/a/a0/Rough_set.jpg">
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our project was to use it as a platform for displaying specific peptides on the surface of <i>E. coli</i>.</p>
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<img src="https://static.igem.org/mediawiki/2014/2/2e/Ag43.jpg" alt="Ag43">
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<p>The preliminary data from the tests showed that the detector was able to differentiate between GFP producing and non-producing bacteria.</p>
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</div>
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<img src="https://static.igem.org/mediawiki/2014/e/e9/Lb_vs_gfp.png">
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<p>Fig.1 Series dilutions to estimate detector sensitivity</p>
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<p>From Fig.1 we can see that the detector can easily distinguish the GFP producing culture from below 1/10 dilutions. Thus we continued on and improved the design by soldering some of the circuit and making it more compact. Then we made more test with the Sender and Receiver bacteria.</p>
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<img src="https://static.igem.org/mediawiki/2014/3/33/Send_rec_vs.png">
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<p>Fig.2 Detector response to Sender and Receiver cultures</p>
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<p>Again the detector is sensitive enough to pick up the GFP in the Receiver.</p>
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<p>For the final test, we made clean overnight cultures of the Receiver and Sender and mixed them in a growing culture. This was done to see after how long the detector is going to pick up the GFP production. Hence, if our diagnostic method is going to work.</p>
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<img src="https://static.igem.org/mediawiki/2014/7/79/Resp_time.png">
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<p>Fig.3 Detector response to QS GFP-producing Receiver and Sender culture, normalised against LB medium</p>
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<p><b>Finally, the simple detector proved capable of detecting the QS culture, thus confirming our design success.</b></p>
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Latest revision as of 02:31, 18 October 2014

Team:Aberdeen Scotland/Parts - 2014.ogem.org



Detector Test Data


The initial concept of the fluorescence detector device had a very rough design. Thus it was built from bits and pieces in order to do a preliminary test and verify its viability.

The preliminary data from the tests showed that the detector was able to differentiate between GFP producing and non-producing bacteria.

Fig.1 Series dilutions to estimate detector sensitivity

From Fig.1 we can see that the detector can easily distinguish the GFP producing culture from below 1/10 dilutions. Thus we continued on and improved the design by soldering some of the circuit and making it more compact. Then we made more test with the Sender and Receiver bacteria.

Fig.2 Detector response to Sender and Receiver cultures

Again the detector is sensitive enough to pick up the GFP in the Receiver.

For the final test, we made clean overnight cultures of the Receiver and Sender and mixed them in a growing culture. This was done to see after how long the detector is going to pick up the GFP production. Hence, if our diagnostic method is going to work.

Fig.3 Detector response to QS GFP-producing Receiver and Sender culture, normalised against LB medium

Finally, the simple detector proved capable of detecting the QS culture, thus confirming our design success.