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Team:Edinburgh/Bacterial
From 2010.igem.org
Bacterial BRIDGEs
Communication capability is a key component of the modern information-driven world. From internet to instant messenger, SMS to telephone, society has developed a large number of technological means for people to keep in constant contact with one another and to exchange information ranging from the trivial to the complex. Even simple speech, and communication of ideas via concepts and words, is a key differentiating factor between human beings and other higher mammals.
What if bacteria such as E. coli were able to communicate via a means more efficient than simple chemical signalling? The creation and sensing of light is not a novel idea in synthetic biology, as evidenced by efforts such as the firefly luciferase reporter developed by Ljubljana 2007 and the photoreceptor submitted by Lausanne 2009. Until now, however, there has not been a concentrated effort to match light production with light reception. The 2010 University of Edinburgh iGEM team has worked to create a standard set of light producing and light sensing BioBricks with which light-based communication can take place.
FORTH stands for Fabricated Organism Reception and Transmission of Heterogeneous light. It establishes a core set of BioBricks that allow synthetic organisms to create light of a determined wavelength upon a specified stimulus, and to activate a specified response when they sense said light.
Our Project
Figure 1: Normalised absorbance spectra of:
a. the lov2 domain of Avena sativa with bound FMN (adapted from Schüttrigkeit et al., 2003).
b. Green light absorbing form of cyanobacteriochrome CcaS from Synechocystis sp. PCC 6803. Adapted from Hirose et al, 2008.
c. Red light absorbing form of phytochrome Cph1-PCB adduct, from Synechocystis sp. PCC 6803. Adapted from Gambetta and Lagarias (2001).
Note that the relative absorbance of each spectrum is normalised to one.
The response of light receptors Cph8, lovTAP and our green light receptor to different wavelengths of light have not been measured. The absorbance spectra of the light sensitive proteins might not exactly mirror their response to light, but should give us a good idea until they are characterized.
In lovTAP, the light sensitive domain of the protein is the lov2 domain from the Avena sativa blue light receptor phototropin. This binds flavin mononucleotide (FMN) which is its co factor. Schüttrigkeit et al. (2003) measured the absorbance of the wild type lov2 bound to FMN (see Figure 3). Since this is the active part of lovTAP, we expect our blue light receptor to have a similar response in vivo. The red light absorbing form of Cph1 is what responds to red light in Cph8. The absorbance of Cph1 was measured by Gambetta and lagarias (2001). Similarly, the absorbance of the green light absorbing form of CcaS, which we are planning on using in our green light receptor, was measured by Hirose et al. (2008).
Table of Contents
- The core repressilator upon which we chose to base our project.
- A description of the work we did on the red light producing component.
- A description of the work we did on the red light sensing component.
- A description of the work we did on the blue light producing component.
- A description of the work we did on the blue light sensing component.
- A description of the work we did on the green light producing component.
- A description of the work we did on the green light sensing component.
- The BioBricks we submitted as part of developing the light producing and light sensing components.
- A summary of what we achieved as part of developing the light producing and light sensing components.
- Our vision of the future of E. coli communication through light, and where we would like to go next.
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References used throughout the section.
