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Team:Imperial College London/M3/Modelling
From 2009.igem.org
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| + | This module is concerned with killing the E. coli bacteria after successful drug protein production and encapsulation. Killing will be activated by an increase in temperatures from 28°C to 42°C. | ||
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The E. coli is killed after it has successfully produced PAH or cellulase, has encapsulated itself in colanic acid and produced trehalose to increase stability of PAH or cellulase during storage. Killing is necessary to make the pill non-toxic or pathogenic, and hence safe for human consumption. This self killing mechanism will be activated by an increase in temperatures from 28°C to 42°C. Therefore, this module will enable us to kill the E. coli after protein production and encapsulation has finished. | The E. coli is killed after it has successfully produced PAH or cellulase, has encapsulated itself in colanic acid and produced trehalose to increase stability of PAH or cellulase during storage. Killing is necessary to make the pill non-toxic or pathogenic, and hence safe for human consumption. This self killing mechanism will be activated by an increase in temperatures from 28°C to 42°C. Therefore, this module will enable us to kill the E. coli after protein production and encapsulation has finished. | ||
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| - | Initially, the lambda cI promoter is repressed by the protein cI that is produced constitutively under the strong promoter J23114. At 28°C, functional protein cI will bind to the lambda cI promoter to repress it. Restriction enzymes DpnII and TaqI will not be produced. | + | <!--Initially, the lambda cI promoter is repressed by the protein cI that is produced constitutively under the strong promoter J23114. At 28°C, functional protein cI will bind to the lambda cI promoter to repress it. Restriction enzymes DpnII and TaqI will not be produced. |
Revision as of 00:14, 15 September 2009
Background
This module is concerned with killing the E. coli bacteria after successful drug protein production and encapsulation. Killing will be activated by an increase in temperatures from 28°C to 42°C.
Goals
We aim to:
- Explore how temperature correlates to restriction enzyme concentration, and see how it affects the population of live cells, so as to characterise the effects of temperature on cell death.
- Develop a model of the number of dead cells as this correlates to our live and dead cells assay. (hyperlink to assay). From this, we can monitor the rate of killing and perform data analysis to obtain the necessary parameters.
- Model for cell death without Dam methylase protection.
In summary, our model will allow us to characterise the effects of temperature on the parallel production of restriction enzymes DpnII and TaqI, that are required to kill E. coli. We can also characterise the extent Dam methylase can protect E. coli against cell death. (not yet done)
<biblio>
- 1 pmid=15652426
- 2 pmid=10235623
</biblio>
