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Team:Macquarie Australia/Project
From 2010.igem.org
Aim
The overall goal of our project is to introduce Deinococcus radiodurans and Agrobacterium tumefaciens bacteriophytochromes into E. coli which have the potential to be used as molecular light switches in response to red and far-red light. Comparison and analysis of the phosphorylated peptides in recombinant E. coli can also be considered in the future. The following are our four main aims which we strive to achieve by iGEM in early November 2010.
Our four main aims
1. Isolate the bacteriophytochrome gene from Agrobacterium tumefaciens and Deinococcus radiodurans.
The expected outcome of this aim is that the Agrobacterium tumefaciens DNA is successfully extracted. It is currently unknown as to the availability of the Deinococcus radiodurans DNA as it is currently being cultured in a Macquarie University facility which we have access to however it is unknown if this can be cultured in the time in which this research is being taken place. If Deinococcus radiodurans cannot be used then the research will proceed with the bacteriophytochrome gene from just Agrobacterium tumefaciens .
2. Transform the bacteriophytochrome gene into Escherichia coli using a vector containing heme oxygenase.
The expected outcome of this aim is that the vector will be successfully cloned into Escherichia coli and will be appropriately selected for by use of antibiotic resistance.
3. Create a biological light switching mechanism using red and frared light.
By using the light sources we expect that the Escherichia coli transformants will turn from blue to green when red and far-red light is absorbed by the colonies respectively. This particular light switching mechanism has not been obtained in an Escherichia coli colony in the scientific community therefore this aim is particularly significant in that a novel function is being genetically synthesised in a model organism which allows for further research into the effects of genetically engineering this switching effect in Escherichia coli.
4. Present the findings at the iGEM conference in Boston MIT, USA in November.
A significant outcome of this research is that it can be applied to the International Genetically Engineered Machine competition, which provides exposure of the research internationally. Also as this is an annual competition this research can be built upon in future years by undergraduate students.
Abstract
Photoreceptors are utilized by almost every organism to adapt to their ambient light environment.
Our aim is to engineer a novel reversible molecular ‘light switch’ within E. coli by introducing a photoreceptor from non-photosynthetic bacteria ( D. radiodurans and A. tumafaciens ).
By cloning the bacteriophytochorome coupled with heme-oxygenase, an enzyme that produces biliverdin from heme, the created colonies are able to respond to red and far-red light environmments.
This novel approach results in the colour of the E. coli ‘switching’ from blue to green.
Our E. coli chameleon will serve as a fundamental ‘bio-brick’ for future applications by providing a simple and photo-reversible switch.
