Difference between revisions of "Team:Newcastle/Proof"

Line 64: Line 64:
 
         <h3>Arabinose Controlled 'Variable Resistor'</h3>
 
         <h3>Arabinose Controlled 'Variable Resistor'</h3>
  
−
         <p>Our aim was to prove that the Arabinose Controlled ‘Variable Resistor’ grew in zinc (II) chloride? When arabinose is present, therefore showing that the SmtA is being produced and binding to the zinc (II). </p>
+
         <p>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 Tokyo-NokoGen iGEM Team in 2011 who used 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), shown by Tokyo-NokoGen, or Zinc (II). </p>
  
−
         <p>For this experiment, we used a plate reader and measured cell survival at various concentrations of zinc (II) chloride?, with and without arabinose present. The plate was set out as seen in Diagram 2. </p>
+
         <p>We took the SmtA part (BBa_K519010) and put it under the control of a pBAD promoter, induced by the presence of L-arabinose. 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>
  
−
        <p>INSERT DIAGRAM OF THE PLATE LAYOUT</p>
 
 
        
 
        
−
        <p>The DH10(α)? cells with the pSB1C3 cells were grown up in liquid culture of LB broth with chloramphenicol overnight at 37°C. The cells were then diluted, using LB broth with chloramphenicol, to an optical density of 0.05 at 600nm. </p>
 
−
 
−
        <p>The zinc chloride was created using XXX of zinc diluted in XXXX? A serial dilution was then made of the zinc chloride in each of the corresponding wells. The cells were then placed in the correct wells.</p>
 
−
 
−
        <p>Arabinose (XXmM) was then added to each well correspondingly. </p>
 
−
 
−
        <p>The results from the experiment can be seen below. </p>
 
−
 
−
        <h4>Results</h4>
 
−
 
−
       
 
−
 
−
      <p> INSERT GRAPHS </p>
 
−
 
−
     
 
−
 
  
 
         <h3>Battery</h3>
 
         <h3>Battery</h3>

Revision as of 16:55, 16 October 2016

Electrically Induced 'Light Bulb'

We aimed to engineer Escherichia coli that it increases fluoresce when an electrical current is passed through the growth medium, via the use of inducible promoters that respond to the heat-stress created by an electrical current.

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

  1. BBa_K1895000 contains a HtpG promoter which is induced by a sigma-factor (σ32). This sigma factor is produced by cells when under different forms of stress. This part also contains a σ32 coding region which should create a positive feedback loop and therefore increase fluorescence.
  2. BBa_K1895006 contains a DnaK promoter which is induced by dnaK, a product of other stress related responses within the cell.

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 Tokyo-NokoGen iGEM Team in 2011 who used 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), shown by Tokyo-NokoGen, or Zinc (II).

We took the SmtA part (BBa_K519010) and put it under the control of a pBAD promoter, induced by the presence of L-arabinose. 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.

Battery

To make a microbial fuel cell we followed the Reading University’s protocol

We sourced the material such as the neoprene gaskets, carbon fibre electrode material, cation-exchange membrane, J-cloth from Professor Ian Head, Dr. Ed Milner and Paniz Izadi from the School of Civil Engineering and Geosciences.

Results

INSERT GRAPHS