Difference between revisions of "Team:Newcastle/Description"

 
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<h2>Our Project </h2>
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<h2>Results</h2>
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<img alt="IDT" src="https://static.igem.org/mediawiki/2016/9/9f/NCLtinker1.png"/>
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        <h3><a href="https://2016.igem.org/Team:Newcastle/Proof/ElectricallyInducedLightBulb">Heat Induced 'Light Bulb'</a></h3>
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<p> The concept of BioBricks was first introduced by Tom Knight at MIT in 2003. His vision was to standardise synthetic biological parts in a similar way to Lego bricks that interlock to form larger constructs. In turn, this has enabled research groups to engineer novel biological systems. Our team has been inspired by Tom Knight’s “Lego-like” approach to synthetic biology.  
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<p> We aimed to engineer <i>Escherichia coli</i> so that it increases expression of a fluorescent protein (sfGFP) when an electrical current is passed through the growth medium, via the use of inducible promoters that respond to the heat-stress response created by the conversion of electrical energy to waste heat.</p>
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<p> We designed two parts (<a href="http://parts.igem.org/Part:BBa_K1895000">BBa_K1895000</a> and <a href="http://parts.igem.org/Part:BBa_K1895006">BBa_K1895006</a>) which respond to the heat-stress in two different ways:</p>
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<li><a href="http://parts.igem.org/Part:BBa_K1895000">BBa_K1895000</a> contains the <em>E. coli</em> <i>htpG</i> promoter. RNA polymerase requires the stress response sigma-factor (&sigma;<sup>32</sup>) to initiate transcription of genes downstream of this promoter. &sigma;<sup>32</sup> is produced by cells when under different forms of stress, one of which is heat. This composite part also contains a modified BioBrick compatible &sigma;<sup>32</sup> coding region (the gene <em>rpoH</em>, <a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1895001">BBa_K1895001</a>) which will create a positive feedback loop to the P<em><sub>htpG</em></sub> promoter, therefore increasing the expression of the downstream reporter gene <em>sfGFP</em> and the  fluorescence of the cell.</li>
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<li><a href="http://parts.igem.org/Part:BBa_K1895006">BBa_K1895006</a> contains the <em>dnaK</em> promoter which, like P<em><sub>htpG</em></sub>, is transcribed via binding of RNA polymerase by &sigma;<sup>32</sup>. P<em><sub>dnaK</sub></em> is placed upstream of the BBa_0034 RBS and BBa_I746916 sfGFP.</li>
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<p>To see our results <a href="https://2016.igem.org/Team:Newcastle/Proof/ElectricallyInducedLightBulb">click here </a></p>
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          <h3><a href="https://2016.igem.org/Team:Newcastle/Proof/VariableResistor">Arabinose Controlled Variable Resistor</a></h3>
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        <p>We aimed to create a biological “variable resistor” by modifying the <i>E. coli’s</i> natural systems to allow for controlled ion uptake. In order to do so, we looked at the work carried out by the <a href="https://2011.igem.org/Team:Tokyo-NoKoGen/metallothionein">2011 Tokyo-NokoGen iGEM team</a> who used the <i>smtA</i> gene from Cyanobacteria and inserted it into a strain of <i>E. coli</i>. SmtA is thought to play a role in preventing heavy metal toxicity by binding excess heavy metal ions such as Cadmium (II), as characterised by <a href="https://2011.igem.org/Team:Tokyo-NoKoGen/metallothionein">Tokyo-NokoGen</a>, or Zinc (II). </p>
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        <p>We took the <i>smtA</i> gene, (<a href="http://parts.igem.org/Part:BBa_K519010">BBa_K519010</a>), and put it under the control of a P<em><sub>BAD</em></sub> promoter, induced by the presence of L-arabinose, making our BioBrick <a href="http://parts.igem.org/Part:BBa_K1895999">BBa_K1895999</a>. This should allow us to control the uptake of zinc ions by adding or removing L-arabinose, resulting in control over the resistance of the LB media.</p>
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<p>To see our results <a href="https://2016.igem.org/Team:Newcastle/Proof/VariableResistor">click here </a></p>
  
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<p>However, we wondered how we could mix electronic devices with synthetic biological devices. Our project therefore involves us replacing some of the traditional electronic components in a circuit with biological alternatives. Like Lego, the circuit will allow synthetic biologists to mix and match bacterial and electronic components to create electro-biological circuits. This approach will represent a foundational advance in the way synthetic biological circuits are designed and implemented.</p>
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<p>We believe that by merging biology, electronic engineering and computer science, our project also holds fantastic opportunities for education. There is a common misconception that all bugs are bad, but our project will demonstrate the benefits of bacteria in a fun and safe manner. </p>
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        <h3><a href="https://2016.igem.org/Team:Newcastle/Proof/MFC">Microbial Fuel Cell</a></h3>
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<p>We aimed to look at different ways of improving the voltage output of a microbial fuel cell. At first we looked at yeast microbial fuel cells with the help of <a href="http://www.ncl.ac.uk/ceg/role/profile/edmilner.html#publications">Dr Ed Milner</a>, <a href="https://www.researchgate.net/profile/Paniz_Izadi">Dr Paniz Izadi</a> and <a href="http://www.ncl.ac.uk/ceg/role/profile/ianhead.html#background">Professor Ian Head</a>, but after <a href="https://2016.igem.org/Team:Newcastle/HP/Gold">talking with PEALS</a> we decided to move away from using yeast and looked at working with <i>E. coli</i> instead. </p>
  
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<p>Synthetic biology is an exciting, and unique field that we want to make accessible to children worldwide. We want the next generation to have a better understanding of what synthetic biology is, and inspire new ways to apply synthetic biology to real world applications. Perhaps our circuits will alter the perception of Genetically Modified Organisms, inspire a new way to generate sustainable electricity, or even work towards creating a computer made entirely from synthetic organisms. </p>
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<p>For inspiration we looked at the <a href="https://2013.igem.org/Team:Bielefeld-Germany/Project/MFC">Bielefeld 2013 iGEM Team </a>. One of the issues we noticed with their design was that their porin overexpression protein was taken from <i>Pseudomonas fluorescens</i> and so the pores size was too large for the <i>E. coli</i> to handle. We changed this by overexpressing <i>E. coli’s</i> natural porin producing genes <i>ompF</i>,<a href="http://parts.igem.org/Part:BBa_K1895004 "> BBa_K1895004</a>. Bielefeld also had issues with cell growth due to the metabolic stress of using a T7 promoter. To improve this part we used a P<sub><i>BAD</i></sub> promoter to allow the cell population to grow before inducing the porin over-expression, <a href="http://parts.igem.org/Part:BBa_K1895005 ">BBa_K1895005.</a></p>
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<h2>Design</h2>
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<p>To see our results <a href="https://2016.igem.org/Team:Newcastle/Proof/MFC">click here </a></p>
  
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Latest revision as of 02:05, 20 October 2016


Results

Heat Induced 'Light Bulb'

We aimed to engineer Escherichia coli so that it increases expression of a fluorescent protein (sfGFP) when an electrical current is passed through the growth medium, via the use of inducible promoters that respond to the heat-stress response created by the conversion of electrical energy to waste heat.

We designed two parts (BBa_K1895000 and BBa_K1895006) which respond to the heat-stress in two different ways:

  1. BBa_K1895000 contains the E. coli htpG promoter. RNA polymerase requires the stress response sigma-factor (σ32) to initiate transcription of genes downstream of this promoter. σ32 is produced by cells when under different forms of stress, one of which is heat. This composite part also contains a modified BioBrick compatible σ32 coding region (the gene rpoH, BBa_K1895001) which will create a positive feedback loop to the PhtpG promoter, therefore increasing the expression of the downstream reporter gene sfGFP and the fluorescence of the cell.
  2. BBa_K1895006 contains the dnaK promoter which, like PhtpG, is transcribed via binding of RNA polymerase by σ32. PdnaK is placed upstream of the BBa_0034 RBS and BBa_I746916 sfGFP.

To see our results click here


Arabinose Controlled Variable Resistor

We aimed to create a biological “variable resistor” by modifying the E. coli’s natural systems to allow for controlled ion uptake. In order to do so, we looked at the work carried out by the 2011 Tokyo-NokoGen iGEM team who used the smtA gene from Cyanobacteria and inserted it into a strain of E. coli. SmtA is thought to play a role in preventing heavy metal toxicity by binding excess heavy metal ions such as Cadmium (II), as characterised by Tokyo-NokoGen, or Zinc (II).

We took the smtA gene, (BBa_K519010), and put it under the control of a PBAD promoter, induced by the presence of L-arabinose, making our BioBrick BBa_K1895999. This should allow us to control the uptake of zinc ions by adding or removing L-arabinose, resulting in control over the resistance of the LB media.

To see our results click here


Microbial Fuel Cell

We aimed to look at different ways of improving the voltage output of a microbial fuel cell. At first we looked at yeast microbial fuel cells with the help of Dr Ed Milner, Dr Paniz Izadi and Professor Ian Head, but after talking with PEALS we decided to move away from using yeast and looked at working with E. coli instead.

For inspiration we looked at the Bielefeld 2013 iGEM Team . One of the issues we noticed with their design was that their porin overexpression protein was taken from Pseudomonas fluorescens and so the pores size was too large for the E. coli to handle. We changed this by overexpressing E. coli’s natural porin producing genes ompF, BBa_K1895004. Bielefeld also had issues with cell growth due to the metabolic stress of using a T7 promoter. To improve this part we used a PBAD promoter to allow the cell population to grow before inducing the porin over-expression, BBa_K1895005.

To see our results click here