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iGEM 2009 Team:Newcastle Bac-Man: The Quest to Sequester
 
We have a problem… ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],Overview So what’s the problem?... … ACCUMULATION
What can our Project do about it? AIMS ,[object Object],[object Object],[object Object]
Our System ,[object Object],[object Object],[object Object],[object Object],Cadmium Intake Cadmium sequestering and sporulation Stochastic Switch Cadmium Sensing Cadmium efflux
[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],Sub-projects Overview
Cadmium Sensing
Cadmium Sensing: What is this sub-project about? ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],Metal  Sensor
Cadmium Sensing: ArsR and CzrA ,[object Object],[object Object],[object Object],[object Object],For the same reason we are using the AND gate – metal sensitive promoters can sense MORE THAN ONE METAL! We know that the CadA promoter contains CzrA binding site Solution: engineer ArsR binding site next to CadA promoter to create cadmium sensing AND gate Metal Sensor  Metals Sensed  ArsR  As(III)  Ag(I)  Cu  Cd  Metal Sensor  Metals Sensed  czrA  Zn  Co  Ni  Cd
Cadmium Sensing: BioBrick Construct AND gate BioBrick ArsR binding sites cadA promoter with CadA binding site RBS Metal  Sensor
Cadmium Sensing: AND gate
Cadmium Sensing: Modelling Metal  Sensor
Cadmium Sensing: Novelty and Importance ,[object Object],[object Object],[object Object],[object Object],[object Object],Metal  Sensor
Stochastic switch
Stochastic Switch: What is this sub-project? ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],Stochastic  Switch
Stochastic Switch: Novelty in design ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],Stochastic  Switch
Stochastic Switch: BioBricks and Devices… EcoRI Xbal Biobrick  Prefix Rfp AccIII BalI hixC hixC hin invertase sigA(pveg) pspac NaeI xlyA promoter NruI SpeI PstI Biobrick  Suffix RBS +  GFP Prefix (ATG) NheI Key Restriction site Invertase site Double terminator RBS ‘ Right facing’ promoter ‘ Left facing’ promoter ,[object Object],[object Object],[object Object],Stochastic  Switch
Stochastic Switch: Modelling ,[object Object],Stochastic  Switch
pSpac XylA Germination activator (FimE) Sporulation  activator  (CinR) pSpac XylA Sporulation  activator  (CinR) Germination activator (FimE) Hin ,[object Object],[object Object],Stochastic  Switch Stochastic Switch: Modelling hixC hixC hin Pveg hixC hixC hin Pveg
Cadmium Sequestration
[object Object],[object Object],[object Object],[object Object],[object Object],Cadmium Sequestration: What is this sub-project about?
Cadmium Sequestration: BioBrick Construct
Sporulation Tuning
Sporulation Tuning: Aims and Novelty ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]
Sporulation Tuning – the sporulation trigger  ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]
Sporulation Tuning: Modelling
Sporulation Tuning: Lab Work and Characterisation
References ,[object Object],[object Object]
Sin (sporulation inhibition) Operon Model ,[object Object],[object Object],[object Object]
Population Modelling
Population Modelling: Aims and Novelty ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]
Population Modelling: How does it work? ,[object Object],[object Object],[object Object]
Population Modelling: Distributed Computing ,[object Object],[object Object],Database Server (Microbase) ‏ Amazon EC2 Cell
Any Questions?

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Newcastle iGEM Presentation 2009

Notes de l'éditeur

  1. Our
  2. Our
  3. The stochastic switch sub project involves a flippable promoter region controlled by the Hin invertase protein. The region will undergo random flipping when the invertase is expressed. The invertase is under the control of 2 variable strength promoters as well as another protein ( sspb from E.coli ) which controls Hin’s degradation rate. This means that we have a 3 way control over the levels of the invertase and so over the flipping rate. Thus our switch is tuneable.
  4. Here is an overview diagram of the entire device that we had synthesised. The diagram shows that for testing purposes we actually replaces the variable strength promoters with inducible promoters pSpac and xylA (induced by IPTG and Xylose respectively). Our degratation controller is induced by Arabinose using the Ara promoter region form the Bacillus ara operon. Overall this means that we are able to test this construct alone without the many other regulatory devices involved. And once characterised we can change the promoters due to the cut sited entered at either side of the inducible promoters.
  5. We need to change these graphs to something that fits with what we have done.