Difference between revisions of "Team:Imperial College/Model"

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          <h3>What is Modelling?</h3>
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          <p2>Constructing and manipulating abstract (mathematical and/or graphical) representations of economic, engineering, manufacturing, social, and other types of situations and natural phenomenon, simulated with the help of a computer system. Also called computer simulation.
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        <h3>Why is Modelling important in biological studies?</h3>
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        <p2>Computer modeling simulates the interaction among many different variables to provide scientists with a general idea of how a biological system may work.</p2>
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                     <p> The PixCell Construct consists of a repurposed version of the soxRS regulon from E. coli, consisting of SoxR and GFP being expressed from either side of the pSoxR/pSoxS bidirectional promoter. pSoxR provides constitutive expression of SoxR. When oxidised, either directly by redox-cycling molecule or by oxidative stress, SoxR binds and activates transcription of GFP downstream of pSoxS.</p>
 
                     <p> The PixCell Construct consists of a repurposed version of the soxRS regulon from E. coli, consisting of SoxR and GFP being expressed from either side of the pSoxR/pSoxS bidirectional promoter. pSoxR provides constitutive expression of SoxR. When oxidised, either directly by redox-cycling molecule or by oxidative stress, SoxR binds and activates transcription of GFP downstream of pSoxS.</p>
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       <p> Using the Golden Gate assembly method, with two inserts; the SOXRS regulon and GFP, which were originally amplified out from the E. coli genome and a storage vector respectively, the construct was assembled. Gel electrophoresis showed that a construct of approximately the right length was produced and the construct was then sequence verified. </p>
 
       <p> Using the Golden Gate assembly method, with two inserts; the SOXRS regulon and GFP, which were originally amplified out from the E. coli genome and a storage vector respectively, the construct was assembled. Gel electrophoresis showed that a construct of approximately the right length was produced and the construct was then sequence verified. </p>
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Revision as of 21:50, 16 October 2018

Modelling



What is Modelling?

Constructing and manipulating abstract (mathematical and/or graphical) representations of economic, engineering, manufacturing, social, and other types of situations and natural phenomenon, simulated with the help of a computer system. Also called computer simulation.

Why is Modelling important in biological studies?

Computer modeling simulates the interaction among many different variables to provide scientists with a general idea of how a biological system may work.

Assembling the Pixcell Constructs

Access All

Description

The PixCell Construct consists of a repurposed version of the soxRS regulon from E. coli, consisting of SoxR and GFP being expressed from either side of the pSoxR/pSoxS bidirectional promoter. pSoxR provides constitutive expression of SoxR. When oxidised, either directly by redox-cycling molecule or by oxidative stress, SoxR binds and activates transcription of GFP downstream of pSoxS.

Figure 1. Illustration of the PixCell Construct

Relevance

The Pixcell construct was designed to test whether we could control the expression of GFP by controlling the oxidation state of SoxR through our electrochemical set up.

Results

Using the Golden Gate assembly method, with two inserts; the SOXRS regulon and GFP, which were originally amplified out from the E. coli genome and a storage vector respectively, the construct was assembled. Gel electrophoresis showed that a construct of approximately the right length was produced and the construct was then sequence verified.

Figure 1. PCR results from the amplification of the entire construct

In order to lower the basal level of GFP expression a degradation tag was added. Therefore, any increase in GFP expression induced by the oxidation of SoxR would be more predominant.

Figure 1. Illustration of the PixCell Construct DegTag

Summary

The Pixcell construct enables us to test whether we can control gene expression by controlling the oxidation state of soxR. The Pixcell construct DegTag enables the system to be more dynamic in that seeing the system switch from on to off is quicker.