IGEM:IMPERIAL/2008/New/Chassis 1: Difference between revisions

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More: http://openwetware.org/wiki/IGEM:IMPERIAL/2008/Prototype/Motility
More: http://openwetware.org/wiki/IGEM:IMPERIAL/2008/Prototype/Motility
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==Light Sensing==
==Light Sensing==

Revision as of 02:45, 8 September 2008

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<html><a href=http://openwetware.org/wiki/IGEM:IMPERIAL/2008/Prototype><img width=50px src=http://openwetware.org/images/f/f2/Imperial_2008_Logo.png></img</a></html> Home The Project B.subtilis Chassis Wet Lab Dry Lab Notebook

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Authors: Wiki team

Editors: Wiki team

Chassis 1

This page offers a brief overview of how B. subtilis meets our main project specifications - to a much higher degree than E. coli! You'll find information on the main mechanisms behind our proposed system, and some very interesting biology too. If you would like even more depth than the outlines below, you are welcome to visit our OpenWetWare pages on those topics - but don't get to engrossed and forget to come back!

Motility

A schematic of the B. subtilis rotary flagellar motor is shown. Motile cells are powered by interactions of the FliG protein with the MotA/B complex (which generates torque). The protein EpsE acts as a molecular clutch to disengage the rotary flagellar motor, leaving the flagellum intact but unpowered. This shuts down motility and facilitates biofilm formation. Fluorescence microscopy photos of B. subtilis show bacterial membranes in red and flagella in green. FliM and FliF are motor proteins [1]

To achieve accurate distribution of our biomaterial microfactories (an affectionate term for our bacteria) we need to be able to exert fine control over their motility. B. subtilis' prime method of getting about is flagellar locomotion; a ring of protein in the cell membrane rotates and is attached to a flagella that extrudes, acting like a long corkscrew propeller to push the cell through its environment.

Where B. subtilis differs from other bacteria is in our knowledge of its mechanism for this movement. A recent paper [LINK] described a clutch mechanism involved in the process; the flagella can be detached from the rotor by expression of a molecule that interacts with the flagella and distorts it so it is disengaged from the rotor protein.

Control over the expression of this protein should allow us very quick control of the bacteria - when we want it to stop we trigger expression of the clutch molecule which halts movement.

To draw a parallel with a car, current synthetic methods of stopping bacteria are akin to destroying the engine (which can take time!) in order to stop it powering the vehicle where the method we hope to take advantage of would be like putting the car into neutral - disengaging the engine from the driveshaft. It's an elegant solution that offers us quick control and also the opportunity for quick reversal (putting the car back into "drive").

More: http://openwetware.org/wiki/IGEM:IMPERIAL/2008/Prototype/Motility

Light Sensing

We have chosen light stimulus as our input for the Bioprinter. Light allows us to generate complex patterns with well defined edges, while gradients in wavelength and intensity will allow us to build up varying concentrations of biomaterial.

Photoreceptors which were considered in our design include rhodopsins, phytochromes, LOV and BLUF. Of these, B.Subtilis contains the blue light sensing protein YtvA which is related to the plant blue-light receptors phototropins. YtvA is a positive activator of the general stress transcription factor σB. Blue and not red light activates YtvA and thus induces σB activity. We could use σB which is expressed upon blue light illumination to activate the production of EpsE and hence control bacteria motility. However, there are other stressors which activate σB, hence we aim to make blue light the main activator of σB by controlling growth conditions.

Biomaterials

http://openwetware.org/wiki/IGEM:IMPERIAL/2008/Prototype/Biomaterials



The next page details some more pros and cons about working with subtilis, as well as an overview of our development of it as a chassis... > The Second Chassis Page >


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