Team:Brasil-SP/Project/ResponseModule

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<h3 align="center">Response Module</h3>
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<h1>Response Module</h1>
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<p><div align="justify">&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;The reporter gene chosen for the output system was the GFP. The main reason for that choice was the simplicity in measuring its fluorescence with the fluorimeter and flow cytometry, but it can be replaced by any other reporter system. In the final development stage of the project we envision a output that do not need to be excited like the GFP does.</p>
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The reporter gene chosen for the output system was the GFP (REMINGTON, 2014). The main reason for that choice was the simplicity in measuring its fluorescenchttp with the fluorimeter and flow cytometry, but it can be replaced by any other reporter system. In the final development stage of the project we envision an output that do not need to be optically excited like the GFP does.
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<img src="https://static.igem.org/mediawiki/2014/c/cd/Resnponse_module.png" width="500" height="370"/>
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{{:Team:Brasil-SP/Templates/Image | image=Circuit_response.png | alignment=center | caption=. | size=500px}}
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Note that the biossensor performs a negative detection. In other words, when the patient is healthy the bacteria will display a green fluorescence, and when in a unhealthy situation the bacteria will not glow. In a more realistic approach, we do not expect our system to have only two intensities (zero or maximum glow). What might happen is that we'll have a more continuous spectrum of intensities, which would make it harder to differentiate between a negative and positive diagnosis. To deal with this problem we are adding a intensity pattern ruler in our [https://2014.igem.org/Team:Brasil-SP/Project/Device  microfluidic device]. Comparing the intensity displayed in the diagnosis chamber with the standardized ones the user will be able to analyse the result correctly
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<!--We have chosen the GFP as a reporter gene to responding our kidney sensing because the facility to found in our laboratory, where all our professors use it in the lab for their researches. We could use another one, like YFP (yellow fluorescence protein) or RFP (red florescence protein) for exemple.</p> 
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<p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;A blood sample from a healthy person, which has low levels of circulating Cystatin C, when in contact with B. subtilis cells triggers the expression of the reporter gene GFP in the biodetector, resulting in high intensity of fluorescence signal. However, when a blood sample from a ill person, wich has high levels of circulating Cystatin C, contacts B. subtilis cells induce the minimum of fluorescence signal, or doesn’t induce the production of GFP. In this way, the intensity of the fluorescence signal show us the diagnostic of the person, if he is sick or healthy, based in a negative signal.</div></p>
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<img src="https://static.igem.org/mediawiki/2014/b/b2/GFP_cell.JPG" width="300" height "auto"><p><strong>High Intensity of GFP</strong></p>
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<h2>Reference</h2>
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#REMINGTON SJ. Green fluorescent protein: A perspective. <b>Protein Science</b>, 2011, 20(9):1509–1519.
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<img src="https://static.igem.org/mediawiki/2014/4/48/No_GFP.jpg" width="200" height="auto"><p><strong>Low/or no Intensity of GFP</strong></p>
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Latest revision as of 00:35, 18 October 2014

TheProjectBRASILSP.png

Response Module

The reporter gene chosen for the output system was the GFP (REMINGTON, 2014). The main reason for that choice was the simplicity in measuring its fluorescenchttp with the fluorimeter and flow cytometry, but it can be replaced by any other reporter system. In the final development stage of the project we envision an output that do not need to be optically excited like the GFP does.

Circuit response.png
.

Indicator.png
.

Note that the biossensor performs a negative detection. In other words, when the patient is healthy the bacteria will display a green fluorescence, and when in a unhealthy situation the bacteria will not glow. In a more realistic approach, we do not expect our system to have only two intensities (zero or maximum glow). What might happen is that we'll have a more continuous spectrum of intensities, which would make it harder to differentiate between a negative and positive diagnosis. To deal with this problem we are adding a intensity pattern ruler in our microfluidic device. Comparing the intensity displayed in the diagnosis chamber with the standardized ones the user will be able to analyse the result correctly


GFP cell.JPG
High Intensity of GFP.

No GFP.jpg
Low/or no Intensity of GFP.

Reference

  1. REMINGTON SJ. Green fluorescent protein: A perspective. Protein Science, 2011, 20(9):1509–1519.


Sponsors

This project is part of iGEM 2014

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