Imperial/Infector Detector/F2620 Comparison
From 2007.igem.org
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<span style="font-size:120%;">'''Key Difference:'''</span> | <span style="font-size:120%;">'''Key Difference:'''</span> | ||
- | *GFP Synthesis - The life of synthesis of ''in vitro'' is about 90 minutes, compared to ''in vivo'' that | + | *GFP Synthesis - The life of synthesis of ''in vitro'' is about 90 minutes, compared to ''in vivo'' that has a life span of days. The differences is that ''in vitro'' has a burst of expression giving a high but short rate of GFP synthesis. ''In vivo'' is a slower rate but more prolonged rate of synthesis. |
*Intial Rate of Synthesis - The initial rate of GFP synthesis is slower for ''in vitro''. This is thought to be due to LuxR levels, which ''in vivo'' are at steady state but ''in vitro'' has to produce the LuxR upon addition of DNA. | *Intial Rate of Synthesis - The initial rate of GFP synthesis is slower for ''in vitro''. This is thought to be due to LuxR levels, which ''in vivo'' are at steady state but ''in vitro'' has to produce the LuxR upon addition of DNA. | ||
Interestingly the values of rate of synthesis are in the same order magnitude of hundreds, this suggesting that the normalisation we are using to compare these chassis is valid. | Interestingly the values of rate of synthesis are in the same order magnitude of hundreds, this suggesting that the normalisation we are using to compare these chassis is valid. |
Revision as of 21:42, 26 October 2007
Summary of Comparison
We thought to compare our in vitro characterisation to the characterisation of [http://partsregistry.org/Part:BBa_F2620 F2620] in vivo with the aim to highlight some of the differences between the chassis and investigate how the constructs characteristics may change between them. The [http://partsregistry.org/Part:BBa_F2620 F2620] is an ideal construct to compare for comparison because of its detailed characterisation in vivo. The construct is the same as the construct 1 that was used for infecter detector, pTet-LuxR-pLux-GFPmut3b. The key results the comparison were;
- The creation of a new unit to allow comparison between in vitro and in vivo chassis.
- That although we are changing the E.coli chassis from in vivo to in vitro the construct characteristic response is independent of the chassis.
These are exciting findings, revealing the potential for the exploration of new chassis and the ability to use constructs in an exchangable mannor.
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Comparison between in vivo and in vitro for rate of GFPmut3b synthesis for 100nM AHL. The in vivo chassis used was the bacterial strain MG1655 and the in vitro chassis was Promega Commercial S30 Cell Extract(60µl)
Key Difference:
Interestingly the values of rate of synthesis are in the same order magnitude of hundreds, this suggesting that the normalisation we are using to compare these chassis is valid. |
Transfer Function
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The graph above shows the transfer function of [AHL] input vs rate of GFP synthesis output. The graph shows the max rate of synthesis for each of the chassis; for in vivo this is the steady state reached after about 30 minutes and for in vitro it is the rate between 60 and 90 minutes which is the maximum rate before the energy limitations of the system cause the rate to drop. The blue line on corresponds to the range of AHL and the response of the in vitro chassis.
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