Latest revision as of 00:35, 18 October 2014

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.

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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 microfluidic device. Comparing the intensity displayed in the diagnosis chamber with the standardized ones the user will be able to analyse the result correctly

High Intensity of GFP.

Low/or no Intensity of GFP.

Reference

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

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-<html>
+{{:Team:Brasil-SP/Templates/Header|headersrc=TheProjectBRASILSP.png}}
-<h3 align="center">Response Module</h3>
+<h1>Response Module</h1>
-<p><div align="justify">&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 fluorescent signals. However, when a blood sample from a ill person, wich has high levels of circulating Cystatin C, contacts B. subtilis cells it doesn’t induce the production of GFP. The difference of fluorescence show us the diagnostic of the person, if he is sick or healthy, based in a negative signal.</p>
+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.
-<p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;The GFP was chosen because the facility to found in our laboratory, we could use another reporter gene, like YFP (yellow fluorescence protein) or RFP (red florescence protein) for exemple.</p>
-<p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;All the blood samples will be filtered through a microfluidics device, extracting just the plasma, that will be analyzed after it contacts with our modified B. subtilis.</p>
-<p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;In this device, we will store B. subtilis as spores, which are activated by rich medium (Luria-Bertani [LB]) before being used in the device. In order to induce the spore formation, the bacteria are incubated in a minimal nutrient medium (Difco Sporulation Medium [DSM]).
-</div></p>
-</html>
+{{:Team:Brasil-SP/Templates/Image | image=Circuit_response.png | alignment=center | caption=. | size=500px}}
+{{:Team:Brasil-SP/Templates/Image | image=Indicator.png | alignment=center | caption=. | size=500px}}
+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
+{{:Team:Brasil-SP/Templates/Image | image=GFP_cell.JPG | alignment=center | caption=High Intensity of GFP. | size=500px}}
+{{:Team:Brasil-SP/Templates/Image | image=No_GFP.jpg | alignment=center | caption=Low/or no Intensity of GFP. | size=300px}}
+<h2>Reference</h2>
+#REMINGTON SJ. Green fluorescent protein: A perspective. <b>Protein Science</b>, 2011, 20(9):1509–1519.
+{{:Team:Brasil-SP/Templates/Footer}}

Team:Brasil-SP/Project/ResponseModule

From 2014.igem.org

Latest revision as of 00:35, 18 October 2014

Response Module

Reference

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