From 2006.igem.org

Making E. coli Glow

Preliminary Design: -Light Sensing Inhibitor- -> -Light emission device-

Light Sensing Inihibitor: [http://partsregistry.org/Part:BBa_I15008 BBa_I15008] , [http://partsregistry.org/Part:BBa_I15009 BBa_I15009] , [http://partsregistry.org/Part:BBa_I15010 BBa_I15010] , [http://partsregistry.org/Part:BBa_R0082 BBa_R0082]

Light Emission Device: (from lux [http://en.wikipedia.org/wiki/Operon operon]) THIS IS WHAT WE NEED TO DO.

Lux Operon

Light Sensor Parts: http://partsregistry.org/Featured_Parts:Light_Sensor and paper : http://www.nature.com/nature/journal/v438/n7067/full/nature04405.html (this paper simply explains how the device works!)

It seems to repress gene expression by having red light inhibit phosphorylation which would activate a promoter. We would replace the LacZ protein coding region with our lux [http://en.wikipedia.org/wiki/Operon operon].

Sequence details:

[http://www.ai.mit.edu/projects/ntt/documents/proposals2000/MIT9904-10/proposal.html According] to Tom Knight, the Photorhabdus luminescens luxCDABE operon that he cloned is NCBI accession number [http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?db=nucleotide&val=155405 M90093]. I checked this sequence against the biobrick restriction enzymes (EcoRI, XbaI, SpeI, PstI, NotI) using the [http://bioinformatics.org/sms2/rest_summary.html Sequence Manipulation Suite]. Results: EcoRI cuts at the ends of the sequence (+2 and -5); XbaI cuts in the middle (+2411); and SpeI, PstI, and NotI do not cut M00093. The question therefore becomes, did Tom Knight's group add or remove restriction sites? We have the DNA, we can test this in lab.