Modified pili expression
We wanted to produce modified pili to primarily bind a variety of metal ions but also with reporters to allow easy verification of protein expression. A two plasmid system was designed where the modified FimH would be expressed from one plasmid and the remaining proteins in the fim operon expressed from the second plasmid. The two plasmids carried compatible origins of replication (pUC and p15A) and different antibiotic resistance genes (AmpR and CmR) to allow for co-transformation in E. coli (Figure 1).
The FimH protein was modified to carry either sfGFP (reporter), 6xHistidine tag (reporter and metal binding) or two different metallothioneins (metal binding) (Figure 2a). See Basic Parts page for information. The Fim operon consisting of six proteins with native RBS (Figure 2b). Four different promoters were chosen to allow investigation of expression in a range of E. coli chassis. See Composite Parts page for information. Together this design gives a collection of parts for modified pili production.
The purpose of this experiment was to determine the effectiveness of using hydrocyclones as a means of heavy sediment filtration. By running a number of experiments, we were able to implement an iterative design process to optimise performance until our hydrocyclones met the industry standard of particle separation. The idea of using hydrocyclones to filter sediment from contaminated water originated through a conversation with one our major stakeholders, Taunton Aquariums.
Our hydrocyclone was developed to prevent the metal binding reactor from being clogged with sediment. This sediment could range from clay, to sand, to gravel and the geometry of the hydrocyclone can be adapted to suit the user's requirements.
Metal Binding Reactor
The metal binding reactor (MBR) is a piece of cylindrical housing apparatus designed to contain our genetically modified bacteria. We conducted experiments to determine the best media to be used in the reactor, and then used design of experiment (DOE) to optimise the conditions inside the MBR to maximise the removal of a chosen solute.
Taunton Aquarium and Plymouth Marine Laboratory had a large influence on the design of our reactor, while results from the MBR were used to inform our model, an interface designed for use by stakeholders, which we trialled with members of Viola Treatment plant.
The purpose of this experiment was to test the effectiveness of using UV as a bactericide to determine if it was suitable to be used within our filtration system as our biosecurity mechanism. The idea originated through our communications with South West Water who employ UV to remove naturally occurring bacteria that are used in their works to digest organic waste. As we are using genetically modified bacteria we felt it necessary to experimentally test its effectiveness.