Team:UCL/Results
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RESULTS
Final experimental data
GOLD MEDAL: IrrE BioBrick for lubricant
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.
SILVER: Lycopene BioBrick
Hypoxia experiment: Inducing oxidative stress
mNARK lycopene enables ecoli growth under Hypoxia conditions
Hypoxia is a condition in which cells are deprived of oxygen due to low concentration of oxygen in the extracellular milieu. In humans, low oxygen levels in the blood affect tissues. Oxygen is essential for diverse cellular functions, such as catabolic and anabolic processes, and low intracellular concentrations have a negative impact on cell functions and survival.
Oxygen deficit can have a severe impact on cellular function, as seen in cell stress. The inability of cells to effectively manage cellular stress over time has been linked to cellular ageing and age-related diseases (Haigis and Yankner, 2010; Poljšak and Milisav, 2012). Cellular stress leads to deregulation of intracellular processes, as both the structure and function of macromolecules are compromised. Furthermore, high amounts of ROS have been implicated in cellular stress and ageing (Poljšak and Milisav, 2012).
We performed an additional assay expressing lycopene under the mNARK promoter, to test if the cells could survive longer under hypoxia-induced stress. E. coli cells transformed with this construct were compared with the wild type TOP10 E. coli (W/T) monitoring growth and division via optical density (OD) at 600 nm, at specific time points – 3 hours and 16 hours following withdrawal of oxygen.
mNARK-Lyco cells had a higher OD compared to the wild type cells. Cells exposed to hypoxia were also compared with the cells that were grown with oxygen. Most cells still survived despite the presence of hypoxia.
Furthermore, with regards to W/T cells, a depletion in the oxygen concentration caused a drop in cell growth and division, as reflected in decreased OD measurements. However, growth and division of mNARK-Lyco-containing cells was maintained in oxygen-deficient environment during the 16-hour time test period.
This shows that our BioBrick construct (mNARK-Lyco) was able to ensure cell growth and division in oxygen-deficient environment.
Inducing oxidative stress through other ways - copper and --
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GOLD:SPY + GFP - pH promotor
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Superoxide Dismutase 3 Biobrick: GFP titration experiment
