IGEM:IMPERIAL/2009/Feedback & Debriefs/Feedback 27 7
Monday 27th July 2009
- Don’t worry about dosage control (will vary on a case by case basis)
- Do not need tags on the protein - but should add one His tag as proof of concept
- Enzyme needs to be exactly the same as human enzyme or will this trigger an immune response?
will this be protease resistant?
- Biobrick protein production for iGEM so we can clone proteins in and out
- For each application
exactly how to make the protein promoter? E.coli strain? CONTROLS! (measure, detect, purify…)
- End alginate research
- Other effects of transcription factor other than colanic acid production
- How do we detect if we are making our Biobrick?
- Visualisation is ok – not quantitative enough though
can we show E.coli coated in calcium phosphate?
- Killing depends on output of module 2
- Promoter leakiness is a PROBLEM
- Think about which RE's to work with
there is a delicate balance between methyltransferases and RE's worried about over expressing methytransferase above what is required RE may get there first and degrade DNA
- Go for an E.coli system and go for a whole RE set
Geoff has EcoRV with RE and methylase on two different plasmids
develop a robust system have one methyltransferase that protects against two different RE’s
- Cutting the gene out from either side after promoter is good enough as it is not functional
- Need some sort of timer/delay mechanism (Biobrick potential)
- External inducers?
- Need to link by experimentation
- Order essentials ASAP (PKU, strain...)
- Need to match specifications to details in each module…
- Gantt chart may help to split areas
- Plan for wet lab (perhaps 2 people in there by the end of this week?)
- Think about experiments we can carry out per module
- Think about modelling (interfaces and some of the modules)
- Remember – think modular!