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Team:KULeuven/23 July 2008
From 2008.igem.org
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Lab Work
Wet Lab
- Stefanie and I prepared wet cultures from the plates with the cells we received from iGEM. We also started on the glycerol stock. Tomorrow we'll continue and make a glycerol stock of all the parts.
The other team started with restriction and ligation of the output.
- Elke, Nathalie and Jan started with cutting and pasting to test the output. We purified R0084, B0032, E0022 and B0015, checked the concentration, made the right digest of it and separated on gel. Only B0032 and E0022 were cut by the enzymes en the right pieces could be cut out for the ligation.
Dry Lab
One big question: is the stop codon in the scar a problem in linking a tag and a protein coding region? The BioBricks of [http://partsregistry.org/wiki/index.php?title=Part:BBa_I712043 Ljubljana] prove not, but is this correct?
Modeling
Modeling wiki has been updated, Output, Cell Death, Inverter, Memory, Filter, Pulse Generator(still need for a parameter check) have been revisited, added graphs of both CellDesigner and Matlab. A pdf file containing the ODE equations and more was made.
I'm affraid we will have to come back on our decision to simply remove the pulsgenerator and put the lactonase production after the filter. The problem is the key-lock system which is leaky in such a way that there is constant lactonase production. Even when this production is very small (low efficiency of rbs for lactonase), an equilibrium will be achieved between lactonase, HSL --> HSL_LuxR-complex and LuxR. The result is a constant amount of LuxR which is still large enough to repress the production of ccdB, so we can never achieve celldeath! Tomorrow we will have to take a look on our pulsgenerator again and see what this component can add to the system. Downside: not all the parameters have been found yet (and Jonas is absent).
