IGEM:IMPERIAL/2009/Encapsulation/Parts/Module3

=Inducible promoter=

Problem
As the encapsulated cell is impermeable/not very receptive to solutes, we need physical means of induction

Possible Solutions
This could be solved in the following ways:


 * light
 * temperature
 * nutrient deprivation.

For temperature, it could be regulated using the following mechanisms:


 * Temperature sensitive lambda cI repressor
 * RNA thermometer
 * Cold sensor hdbB

Our solution
Using the temperature sensitive lambda cI repressor

BBa_K098995 by Harvard 2008 (on our 2009 distribution plate) http://partsregistry.org/Part:BBa_K098995

This is a thermosensitive cI (clts) inducible system driven by a strong promoter.



For strong promoter J23114 Variant RFP= 256 au

BBa_R0051 is a standard lambda cI regulated promoter. This system is leaky so we have decided to make our own or obtain a commercially available thermoinducicble system.

=Restriction Enzymes-Methylase system=

Dpn1

 * blunt cutter
 * recognises GATC

Taq1

 * staggered cutter
 * recognises TCGA

Dam Methylase
=Weak constitutive promoter=

To maintain a basal level of methylation

Using BBa_J23103 variant RFP= 17 au A weak promoter in the popular constitutive promoter family. http://partsregistry.org/Part:BBa_J23103

=Overall design=

The schematic below shows the overall design of the M3 system:

=Useful reading=

| Harvard Thermoinducible cI system


 * 1) mod1 pmid=5274442


 * 1) mod2 pmid=10659857