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

Monday 27th July 2009
Module 1

will this be protease resistant? exactly how to make the protein promoter? E.coli strain? CONTROLS! (measure, detect, purify…)
 * 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?
 * Biobrick protein production for iGEM so we can clone proteins in and out
 * For each application

Module 2

can we show E.coli coated in calcium phosphate?
 * 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

Module 3

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 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
 * Killing depends on output of module 2
 * Promoter leakiness is a PROBLEM
 * Think about which RE's to work with
 * Go for an E.coli system and go for a whole RE set
 * Failsafe
 * 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!