IGEM:IMPERIAL/2007/Projects/chassis/Design

• Introduction
• Specifications
• Design
• Notes

Phase 1 - Intial Testing

Intialy testing of various simlpe constructs to confirm there is gene expression in vitro and in veso, the simlpe constructs are:

	Test Constructs
Constitutive E.coli Promoter	Ptet promoting GFP
Constitutive T7 Promoter	PT7 promoting GFP
Inducible E.coli Promoter	PBad promoting GFP
Inducible E.coli Promoter	Ptet promoting LuxR, Plux promoting GFP

Firstly we will test the promoters in vitro system. We will be simply testing whether the promoters actually work so we are not interested in a lot of details at present. The promoter construct plus any inducer required (if the promoter is inducable) will be mixed together and the levels of fluorescence will be monitored over time.

We preferably want to use a constitutive promoter for Cell by Date. If all constitutive promoters are found working then we will need to chose one of them. The dicision can be made based on their levels of activity (the higher, the better) and their operating temperature range. This needs to be as close as possible to the CBD operating range of 4^oC - 37^oC

For Infector Detector, only the Lux promoter can be used so it is needed to work in vitro.

Phase 3 -Validation device construct

The two simple constructs will then be implemented in the two project applications of cell by date and infecter detector.