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Team:KULeuven/15 July 2008
From 2008.igem.org
Contents |
Lab Work
Wetlab
Today was our first day in the lab. We prepared LB-medium, Ampicilline,... Everything is now ready for the serious labwork: from tips to cleaning our benches. We did a first attempt to punch out a part of the registry. Then we transformed competent cells (DH5alpha) with the part. We started with the input device: M30109. Stefanie was very excited to pour plates!
Dry lab
Discovered that the LVA tagging T7 RNA polymerase is a no go as the C-terminus is part of the active site of the enzyme. We'll have to do N-terminal tagging, this could work, see the crystal structure of T7. See the link under literature on this wiki. Lon-mediated degradation of lambda excisionase (Xis) and E.coli UmuD gene product. 30-40 amino temrinal AA of UmuD should do the trick.
Modeling
Parameters, parameters, parameters, ... and Matlab, aah... (total smiley face)
Meeting
Present: All students, IT, KM, JW, JR, BDM, AC
Some remarks we have got on our project:
- They showed us where we could find the right E.coli strains
- There could be a problem with photobleaching of eCFP
- The production of eCFP could also be lowered by the low efficiency of the RBS
- Will there be enough ribokey? (Hanne says: ENHANCER!)
- Enough lactonase?
- Maybe we should put tetR under control of his own promotor and lift out of the cell death construct
- We can inhibit our system exogenous by adding lactonase to our medium
- We should use different ori's for our different plasmids, perhaps we 'll have to link some devices
- The engineers should make a simple scheme that indicates which parameters are important for the sensitivity of our system
