Team:Cambridge
From 2010.igem.org
Abstract
We placed genes from fireflies and bioluminescent bacteria into E.coli. Codon optimisation and single amino acid mutagenesis allowed us to generate bright light output in a range of different colours. Future applications include and quantitative biosensors and biological alternatives to conventional lighting.
Multimedia
If you want a break from hard-core science, check out our Gibson Assembly music video.
You can also see view videos of our bacterial bubble lamp and project overview.
Tools
The Project
Project Firefly
Over the course of the summer we have built a number of BioBricks to allow bioluminescence. We used genes originally found in fireflies (Photinus pyralis and Luciola cruciata).We adopted a number of strategies to extend the use of firefly luciferase:
- Codon optimisation for increased light output
- Use of a luciferin regenerating enzyme.
- Mutagenesis to create a number of different colours
Project Vibrio
We complemented these firefly systems, which require the addition of the substrate luciferin, with light producing systems from Vibrio fischeri. We believe we have created the first BioBrick to emit light in normal E. coli strains without the addition of any external substrate.Tools
Over the summer we constructed a number of tools to assist the synthetic biologists of the future:
- Gibthon allows the user to enter a series of BioBrick or GenBank IDs in a specific order and computes the appropriate primers for Gibson Assembly.
- BioBrick → GenBank allows parts from the registry to be downloaded in .gb format, making them compatible with a wide range of biological software.
- The E.glometer is a cheap, easily built, piece of electronics for measuring bioluminescence. It allows scientists without access to expensive plate readers to measure bioluminescence.