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Team:Cambridge/Notebook
From 2009.igem.org
Contents |
iGEM Thought-shower
Monday 12 July
Each team member researched their own ideas for our project
Tuesday 13 July
Afternoon presentations - possible projects included
- Bacto-Stat
- Counterfeit Bill Detector
- Traffic light-obedient bacteria
- Light-induced pigment production
- Predator prey bacteria + a parasite
- Modelling pesticide resistance
- Galvanotaxis
- Wave-pulse bacteria
- Ai2 quorum sensing system
Conclusions
Decided to concentrate on a pigment output. Wednesday plans involve discussing ideas for pigments, following the same meeting pattern used on Tuesday. Ideally the pigment output would be a new registry part which could then be connected to any input system
Wednesday 14 July
Investigated possible outputs, with a kind of bacterial printer in mind.
Colour Wheels
- Primary school style: BLUE, YELLOW, RED
- True colour wheel: YELLOW, CYAN, MAGENTA
- Autochrome: ORANGE, GREEN, VIOLET
Viable Pigments
- In an ideal system, we would have lots of pathways making different coloured pigments from a common precursor. One can dream...
- Carotenoids - RED, ORANGE, YELLOW
- Biobricks exist for part of the system
- apparently you can go from yellow to white (cool)
- Pseudomons aeruginosa - RED, GREENISH/BLUE
- Pyocyanin is greenish/blue, can be synthesized from chorismic acid, or more simply, from phenazine-1-carboxylic acid (PCA)
- Knocking out one of the genes between PCA and pyocyanin leads to the production of a red pigment
- Chromobacterium violacein - VIOLET, CYAN
- Violacein is a violet pigment
- A precursor is cyan.
- BROWN
- melanin - easily attainable
- anyway we could make black?
Follow ups for tomorrow
- Possible inputs
- Population control
- bacterial chlorophyll?
Thursday 15 July
Hoping to start wet work on Monday!
Pigments
- Duncan has orange and brown bacteria we can start to work with. The melanin gene has been sequence, has a restriction site we would need to remove to submit it to the registry.
- We have the genes for violacein! However there seem to be several internal restriction sites within the plasmid that would need to be removed, with three being withing the violacein operon.
- Contacted the authors of a paper to get the genes for pyocyanin biosynthesis. Two of them have forbidden restriciton sites, so we'll need to figure out how to remove them.
Inputs
- explored the idea of using common repressor / inducer systems - arabinose, lac repressor, and tet repressor - to control pigment production
- scourged the registry for lots of different inducible promoters
Population Control
- Crispian proposed a growth control dependency pathway that would make the growth of each type of bacterium (red colour-producing, blue colour-producing, yellow colour-producing, for example) dependent on an other using HSLs
Friday 16 July
- found where to get PCA, pyocyanin precursor
- did an inventory of supplies
- familiarized ourselves with protocols and planned for next week's wet work
- Thinking about a team logo
Current plans for the wet work involve starting transformation of the bacteria in order to view the pigment colours and intensities. For this we will need to create competant cells and then use Duncan's plasmids to transform the violet and brown and the biobrick registary parts to transform the red and orange.
The Project
Monday 20 July
Finalized our project into a few objectives
- Pigment production
- Transform E. coli to produce pigment
- Hook up the pigmentation genes to inducible promoters
- Toggle switch
- build, test a toggle switch
- Hook up pigmentation genes under inducible promoters to the toggle switch
- Improve on previous iGEM projects
- arsenic sensor?
- bacterial photography?
- Modelling
- toggle switch
- Growth rate regulation
Goals for Wet Work
- Inducible pigment production integrated with a toggle switch
- Possibly attach the arsenic bio-sensor to our green/red pigment output
