Team:TU Delft/11 August 2010 content
From 2010.igem.org
Alkane degradation
The colony PCR run overnight on colonies possibly containing BioBrick 020A were checked on gel, but unfortunately none of the products had the correct length (2482bp). We will try to make the BioBrick again tomorrow.
Lane description
# | Description | Primers | Expected length (bp) | ✓ | ✗ |
L | SmartLadder | n/a | n/a | n/a | n/a |
1-16 | 020A | G00100 + G00101 | 2482 | none | all |
L | SmartLadder | n/a | n/a | n/a | n/a |
Modelling of hydrocarbon sensing
To characterize the hydrocarbon sensing system (AlkS and promotors), a model has to be build that describes the GFP and RFP expression that will be measured with the plate reader. The AlkS protein can bind to various promotors (PAlkS1, PAlkS2, PAlkB) and hydrocarbons. This results in the following protein state diagram:
Part of the constants will have to be assumed, and hopefully some can be estimated from plate reader expression data.
Matlab implementation of this model, and some initial guesses for constants results in the following graphs:
It shows that the model correctly maintains a basal level of AlkS when there are no hydrocarbons (HC), and increases to a high level when HC are added.