Kinetoplastida parasites infect humans worldwide causing high morbidity, and still there are many mysteries regarding the parasites’ metabolism. During a summer research experience our lab studied kinetoplastid metabolism, so we cloned DNA that would enable live <i>Trypanosoma brucei</i>, a species of kinetoplastida, to express fluorescent protein biosensors capable of detecting FBP. Trypanosomes prepared to express fluorescent protein biosensors were set to incubate in PBS plus 10 mM glucose as well as a PBS minus glucose control, and then they were run through a flow cytometer. The analyzed flow cytometry data revealed that 3% of the trypanosome population could fluoresce in the presence of FBP. This small fluorescent population can be sorted and raised, which would benefit the lab’s effort to continue monitoring metabolites of glycolysis in trypanosomes as they react to various drugs. With the help of my mentor, a medium for starving trypanosomes, RPMI-Theta, against PBS also tested to determine if it should be used in future experiments; the results indicated RPMI-Theta promotes parasite metabolism and therefore should not be used.

Detecting FBP in live trypanosomes using fluorescent protein biosensors

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