SNOWDRIFT/our idea
From 2009.igem.org
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+ | <p><span style="font-weight:bold; font-size:200%; color:#0000cc;">Our Idea</span></p> | ||
We intend to simulate this system in vivo using engineered E.coli. The scheme is as follows: | We intend to simulate this system in vivo using engineered E.coli. The scheme is as follows: | ||
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Strain 2: (Co-operator Strain): Contains our construct. | Strain 2: (Co-operator Strain): Contains our construct. | ||
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+ | <p><span style="font-weight:bold; font-size:150%; color:#0000cc;">The Construct</span></p> | ||
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+ | [[Image:Constructsnow.JPG|center|500px]] | ||
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+ | The above construct contains a constitutive promoter (J23118 ), an export tag (K233307) in combination with enzyme B-galactosidase and a downstream reporter GFP. | ||
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The co-operators in the population have the ability to synthesize B-galactosidase enzyme constitutively. This enables them to metabolize intracellular lactose into glucose and galactose which are finally converted to usable forms of energy (ATP) which is essential for cell survival, growth and reproduction. The defectors in the population ( LacZ -) do not possess this ability. Thus they must depend upon other carbon sources. This is made available to them due to the presence of the export tag attached to the B-galactosidase enzyme. The export tag facilitates the diffussion of B-galactosidase into the extracellular environment via the TAT-export pathway. This causes the extracellular accumulation of B-galactosidase. The enzyme, active in the cell environment, will cleave lactose molecules into glucose and galactose, which freely diffuse throughout the medium. These molecules can be taken up by the defectors, "free of charge". Thus the growth of defector cells will be supported by the "co-operative" act of B-galactosidase secretion. If the population of co-operators is high, large amounts of extracellular B-galactosidase will accumulate leading to high amounts of extracellular glucose. This facilitates the growth of other defectors and co-operators as well. However, since the defectors do not have to pay the metabolic "cost" of producing and exporting the enzyme, they will have a slight growth advantage over the co-operators. This will lead to rise in defector numbers. However, if the number of defectors becomes too large, the co-operators can no longer produce enough B-galactosidase vis-a-vis glucose to support the entire population. The population would then begin to decline, and would favour the co-operators (who can use the intracellular lactose for their own growth) till the balance is restored. | The co-operators in the population have the ability to synthesize B-galactosidase enzyme constitutively. This enables them to metabolize intracellular lactose into glucose and galactose which are finally converted to usable forms of energy (ATP) which is essential for cell survival, growth and reproduction. The defectors in the population ( LacZ -) do not possess this ability. Thus they must depend upon other carbon sources. This is made available to them due to the presence of the export tag attached to the B-galactosidase enzyme. The export tag facilitates the diffussion of B-galactosidase into the extracellular environment via the TAT-export pathway. This causes the extracellular accumulation of B-galactosidase. The enzyme, active in the cell environment, will cleave lactose molecules into glucose and galactose, which freely diffuse throughout the medium. These molecules can be taken up by the defectors, "free of charge". Thus the growth of defector cells will be supported by the "co-operative" act of B-galactosidase secretion. If the population of co-operators is high, large amounts of extracellular B-galactosidase will accumulate leading to high amounts of extracellular glucose. This facilitates the growth of other defectors and co-operators as well. However, since the defectors do not have to pay the metabolic "cost" of producing and exporting the enzyme, they will have a slight growth advantage over the co-operators. This will lead to rise in defector numbers. However, if the number of defectors becomes too large, the co-operators can no longer produce enough B-galactosidase vis-a-vis glucose to support the entire population. The population would then begin to decline, and would favour the co-operators (who can use the intracellular lactose for their own growth) till the balance is restored. | ||
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There are several confounding factors, which need to be considered while postulating that such a steady state will actually be acheived in our system. One assumption is that the conditions are non-limiting, which may not be the case in our system. As the population grows, the levels of lactose would go down at an ever-increasing pace. This would change the final dynamics and the stable ratios obtained. Again, such stable ratios are obtained after several generations of balancing selection. This would mean that the experiment needs to be carried out under invariant conditions for several generations before the ESS is actually acheived. These and other complications are considered in the modelling section. | There are several confounding factors, which need to be considered while postulating that such a steady state will actually be acheived in our system. One assumption is that the conditions are non-limiting, which may not be the case in our system. As the population grows, the levels of lactose would go down at an ever-increasing pace. This would change the final dynamics and the stable ratios obtained. Again, such stable ratios are obtained after several generations of balancing selection. This would mean that the experiment needs to be carried out under invariant conditions for several generations before the ESS is actually acheived. These and other complications are considered in the modelling section. | ||
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<html> | <html> | ||
+ | <p><a href="https://2009.igem.org/SNOWDRIFT/Proof_of_concept | ||
+ | "><span style="font-weight:bold; font-size:150%; color:#0000cc;"><u>Proof of concept</u></span></a></p> | ||
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</html> | </html> |
Latest revision as of 02:32, 22 October 2009