Imperial/Cell by Date/Testing

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<center> [https://2007.igem.org/Imperial/Cell_by_Date/Implementation << Implementation ] | Testing/Validation https://2007.igem.org/Imperial >> Home | ]</center>

Revision as of 14:38, 22 October 2007


Testing and Discussion

Investigation properties of system under isothermal scenarios

Isothermal Data Arrhenius Plot

In order to investigate the properties of our system we began by looking at isothermal scenarios in order to determine primarily the activation energy of our sytem following Giannuzzi's work with beef. Again this is useful because as per Taoukis' research for a TTI to function correctly its activation energy needs to be close to the target product. In Figure 1 above we see that under isothermal conditions a steady rate of fluoresence was observed, corresponding to a steady rate of protein production. This rate increase with temperature as per our predictions as protein production is linked to the kinetics of translation and so temperature. We have been able to extract the rate of protein production for range of temperature and made an arrhenius plot to further explore translation kinetics. A strong linear correlation in the log plot seems to suggest that an arrhenius type dependance on temperature seems to fit the translation mechanism. From this we have been able to calculate the activation energy as 1.5kJ/mol.

In addtion to calculating the activation energy through these experiments we have also been able to determine the operating range of our system. Our system seems to 'turn off' at around 4 degrees C. At 8 degrees with have minimal expression which then increases to a substantial expression at 37 degrees. Unfortunetly we have not been able to use a visible reporter and so have no idea whether the construct can in fact produce a visible signal. In terms of lifespan we have had major problems with evaporation.

Investigation System properties under Dynamic Temperature Conditions

CBD Steps.pngCBD Steps.png


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Validation and Conclusion

Property
Value
Achieved ?
Inputs
Isothermal conditions between 0 & 40 C
No
Dynamic conditions eg. steps & ramps
hmm
Outputs
System should give a visual signal to indicate level of thermal exposure where beef is off
No we were unable to use a visual reporter
Activation Energy
System needs to have an activation Energy 30 +/- 20 kJ/mol
No activation energy of system was calculated to be 1.5kJ/mol
Health Regulations
System must not be living replicating bacteria
Yes through use of cell free chassis
Response Time
System needs to have a response time under 1 hour
hmm tbd
Lifespan
System must have a shelf life of 7 days
No with current packaging methods evaporation meant that our system 'died' after 2 days



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