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Team:Newcastle
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Revision as of 13:28, 24 July 2013
BareCillus
Our project focuses on the creation and potential applications of L-forms of Bacillus subtilis. L-forms are bacterial cells that are able to propagate and grow without a cell wall. We propose that L-forms can be used as a novel chassis for synthetic biology.
We are creating a BioBrick to facilitate the switching of Bacillus subtilis cells from rod-shape to L-form allowing future iGEM teams to work with this as a chassis. This BioBrick will be under the control of a xylose inducible promoter allowing L-form cells to return to rod-shape when required.
One of the many applications possible with L-forms is genome shuffling. It is significantly easier to fuse bacteria without cell walls interfering with the process. Fusion of L-forms will cause genetic recombination between cells, forcing bacteria to reproduce sexually, and can be used for directed evolution. We will create BioBricks encoding green and red fluorescently labelled Hbsu’s (non-specific DNA binding protein) in order to tag bacterial chromosomes, and observe this recombination.
We aim to put L-forms into single cell wide microfluidics channels with differently shaped end chambers. We will then observe whether the L-forms will grow and take up the shape of these chambers due to their lack of cell wall.
L-forms can exist in plants in a symbiotic relationship with the plant providing an osmotically suitable environment. In return, L-forms can confer benefits to their host including reducing the rate of fungal infection. We plan to grow GFP labelled L-forms inside plants to visualise this relationship. Future L-forms could be engineered to supply nutrients to their host plant, potentially increasing crop yield in low fertility soil. L-forms are osmotically sensitive, giving the ethical advantage that they would lyse if they escaped from their host plant into the environment.
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Training Week
Training Week
The team spent the first week of iGEM learning different basic lab techniques and modelling principles. Everything was documented and written up as a "Starter's Guide" to Synthetic Biology which can be found by clicking this box.
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L-forms
L-form
An L-form is a bacterium that has no cell wall. Bacterial morphology is determined by the cell wall, and so their morphology differs from the strain of bacteria from which they are derived, giving rise to a variety of cell sizes. The cell wall is important for cell division. Binary fission is a highly conserved mechanism required for proliferation of almost all cells. Due to the absence of the cell wall, L-forms are easily transformed, so we propose to use inducible L-forms of Bacillus subtilis as a novel chassis.
We are working on four themes which include: Shuffling, Recombination & Endosymbiosis; Introducing and detecting L-forms in Plants; Shape-shifting; Investigating two-component systems in L-forms. Details of the individual themes can be found under "Projects" on the main menu
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