Our Project

Our project focuses on the creation and applications of L-forms: bacteria that can grow and divide without a cell wall. We propose that L-forms can be used as a novel chassis for synthetic biology.

We have created a BioBrick that switches Bacillus subtilis cells from rod-shape to L-form. This introduces an inducible promoter to control cell wall synthesis, allowing L-form cells to return to rod-shape when required.

We will be using L-forms to attempt 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,causing sexual reproduction, allowing directed evolution.

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 a symbiotic relationship in plants which we will visualise by growing GFP labelled L-forms inside seedlings. L-forms could be engineered to supply nutrients to their host. L-forms are osmotically sensitive, giving biosecurity that they lyse if they escape from their host plant.

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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-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