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Applications of BBa_K729001

UCL iGEM 2016 proved that IrrE could be repurposed for assisting bacterial growth within lubricant.

IrrE Concept

IrrE is a part (5) from a former UCL iGEM team, where, when E. coli was transformed, protected against salt, oxidative and thermal shock. In our case we would see if E. coli transformed with IrrE is better adapted to living in lubricant than wild type E. coli. If so the IrrE transformed E. coli cells could be used as a chassi to introduce pathogen detecting and reporting systems into. Originally IrrE protects against radiation as it is a protein originating from Deinococcus radiodurans.

Experimental Design

The experiment to be conducted is to see whether IrrE transformed E. coli can more readily survive in lubricant than wild type E. coli. To test this firstly wilt type E. coli will be introduced into different concentrations of Superdrug, head quartered in Croydon, Surrey, (Code: 380900) lubricant and LB and be grown over several hours. Then the same will be done with the transformed cells and we will check whether there is a significant difference between the growth curves of the different cells in different concentrations.

Diagram to show different growth of wild type E.coli vs. IrrE E.coli in 40% Lubricant 60% LB using Absorbance at 600nm

From the data shown in the graph above it clearly shown that the E.Coli transformed with IrrE grows better in the 40%Lubricant/60%LB solution in direct comparison with the Wild type E.Coli, which hardly grew at all. This data thus suggests that with the IrrE the E.Coli is better adapted to living in, which would allow further studies being conducted on the maximum concentration of Lubricant it can be grown in. Furthermore once that has been determined the pathogen detecting aspect of the concept can be realised. Thus overall this experiment has allowed us to show that IrrE increases the growth of the E. Coli in the Superdrug Lubricant containing: purified water, glycerine, Carbopol 940, Triethanolamine and Sodium Butyl Paraben. This supports the previous experiments that concluded that IrrE allows for better growth in saline conditions[iv], except this time in Superdrug Lubricant.

For further conceptualisation and experimentation it should be taken into consideration that within a commercialisable product the bacteria will not be under ideal growth conditions and the nutrients will be a limiting factor, especially when considering that the lubricant concentration will have to be a lot higher than in this experiment.

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