IGEM:IMPERIAL/2009/Feedback & Debriefs/Feedback 27 7

Monday 27th July 2009

Module 1

• 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…)

Module 2

• 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
• Experiment

can we show E.coli coated in calcium phosphate?

Module 3

• 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

• Failsafe

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

Linking Modules

• Need some sort of timer/delay mechanism (Biobrick potential)
• External inducers?
• Need to link by experimentation

General

• 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!