IGEM:Imperial/2010/Output module

Effector 1 (Protease)	Effector 2 (Dye,Enz)	Pigment biosynthetic pathways
transcr sigma 54	2 colourless	Bilins
Activation (phosphorylation)	Enz-in-pathway short pathways ensure - colourless -> colour	Anthocyanins short pathways ensure - colourless -> colour
Target proteases to use not many AA non-toxic can work in E.Coli quantize speed, efficiency	Protein scaffold	Fret pairs as back up for effectors
2C DNA binding prot release

This review article has some useful information on FRET.

James- some suggested papers:

1. Kerppola TK. Complementary methods for studies of protein interactions in living cells. Nat Methods. 2006 Dec;3(12):969-71. DOI:10.1038/nmeth1206-969 |
2. Wehr MC, Laage R, Bolz U, Fischer TM, Grünewald S, Scheek S, Bach A, Nave KA, and Rossner MJ. Monitoring regulated protein-protein interactions using split TEV. Nat Methods. 2006 Dec;3(12):985-93. DOI:10.1038/nmeth967 |

GFP as output

• In vivo and in vitro protein solubility assays using split GFP
• Monitoring of conformational change in maltose binding protein using split green fluorescent protein
• Protein Splicing-Based Reconstitution of Split Green Fluorescent Protein for Monitoring Protein−Protein Interactions in Bacteria: Improved Sensitivity and Reduced Screening Time
• A Fluorescent Indicator for Detecting Protein−Protein Interactions in Vivo Based on Protein Splicing