Team:British Columbia/DSZNotebook

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To obtain a control for our project which distributes the DszABC operon amongst three different auxotrophs, we needed to show the rate of desulfurization (removal of dibenzothiophene) when the entire DszABC pathway is transformed into a single host. To perform this experiment, we grew a culture of E.coli containing the optimized plasmid for the DszABC pathway cloned from Rhodococcus erythropolis IGTS8. This construct was obtained from Dr.Keasling and was the source for our DszC biobrick.

An overnight culture for this E.coli cloned with the DszABC operon was grown overnight in 5mL culture of LB. We inoculated the overnight culture in 200mL culture of LB in a 500mL flask so that we have a starting O.D600 is 0.05. Prior to inoculation, dibenzothiophene was dissolved in DMSO at a concentration 100ppm. This was added to the 200mL culture such that it contains 2% DMSO. For a control, we inoculated an E.coli K12 strain without the plasmid, which should not contain the necessary enzyme of biodesulfurization.

Next, we incubated the