Team:Goettingen/Project

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Contents

Overall project

Tell us more about your project. Give us background. Use this is the abstract of your project. Be descriptive but concise (1-2 paragraphs)

The commonly used bacterial model organism Escherichia coli possesses lots of beneficial traits like, for example a short generation time or easily accessible genetic manipulation tools. Most E. coli strains used in laboratories do not exhibit high motility. This is due to a reduction of the bacterial flagellum, which is the crucial element for a motile cell. Our project aims to engineer the bacterium to generate a genetically modified strain that is able to move at a higher velocity. Using directed mutagenesis techniques of motile E. coli strains their swarming ability should be enhanced.

The flhDC operon codes for the master regulator FlhD4C2. This protein complex acts as a transcription factor regulating flagellum biosynthesis and assembly. The increased expression of the master regulator via the introduction of the operon into our E. coli strain leads to the production of additional flagella and thus to hypermotility compared to usual laboratory strains.

To investigate the mobility special “swimming plates” were developed and optimized. These plates contain only a small amount of agar to allow observable movement of the bacterial cells. Furthermore, a second assay introducing a nutrition gradient should trigger a directed movement towards a particular substance. This behavior is called chemotaxis. With these plates in hand, we are able to design an effective motility-selection method. The recognition of these substances is accomplished by chemoreceptors. We work with the receptor TAR that specifically recognizes aspartate. The ligand-binding domain of each subunit consists of two short and two long α-helices to form a four-helix bundle. We aim to change the binding specificity of the ligand-binding pocket by using directed mutagenesis techniques. The establishment of a mutation library will hopefully result in the recognition of novel substances.

In a global perspective, we want to couple substrate sensing (chemotaxis) with motility. Our engineered bacteria could then sense molecules like pollutants or medically relevant substances, like for example cancer markers and move directed and fast towards these.By this means, the targeted substance cannot only be detected but eventually also rendered harmless.

Project Details

Part 2

The Experiments

Part 3

Results