All 1 entries tagged PhD Synthetic Clock
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November 14, 2005
Writing about web page http://blogs.warwick.ac.uk/ulrichjanus/entry/todo_1/
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Results of the meeting with Markus and Isabelle last Friday (11.Nov). Email to get the E.Coli strain used by Atkinson 2003 is under way..
Motivating or guiding Question
- How accurately can synthetic systems be described by mathematical models?
- How well can specific parameters of the in vivo system be changed?
- Review the theoretical model (kinetics and structure) to make it more realistic.
- Identify relevant parameters to move system into the region of sustained oscillations.
- Achieve this parameter shift by mutagenesis of transcription factors by error prone PCR.
- Compare responses of theoretical and in vivo system.
- Model predictions can be improved by using more realistic kinetic description of the regulation events.
- Mutagenesis by PCR allows are more precise manipulation of model parameters than the method used previously.
Schedule of the Experimental Work:
- Check if the strains are still rythmic, i.e. if the previous work can be reproduced (~ 1mth.)
- Intoduce a luciferase reporter gene to measure clock function via luminescence assays (~ 1mth.)
- Test, if cells can be grown on agar plates rather than in liquid cultures
- mutagenise the sequence of relevant transcription factor to change chosen model parameters by erroneous PCR (~ 2mth)
- Identify mutated colonies by changed clock behaviour caused by changes in the affinites of the transcription factor (~ 4mths)
These are 8mths, make it 12mths.
about the advisor(s):