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UBC Theses and Dissertations

Cloning and expression of organic cation transporters (ORCT and ORCT2) from the fruit fly Drosophila melanogaster Meigen Matier, Brieanne Joelle


Organic cations include endogenous metabolites, and xenobiotics (drugs, pesticides, environmental toxins), that must be effectively eliminated in order for organisms to survive. The midgut and Malpighian tubules of insects have been shown to play a role in the active transport of organic cations. Two putative organic cation transporters (OCTs) were cloned from adult Drosophila melanogaster. Phylogenetic analysis indicated that these OCTs exist in an insect specific clade separate and equally divergent from identified vertebrate OCT isoforms. Gene expression patterns for these D. melanogaster transporters were determined using quantitative real time polymerase chain reaction (qPCR). D. melanogaster genes were found to be differentially distributed across the Malpighian tubules and the midgut. The isolated ORCT protein was transiently expressed in Sf9 insect cell lines. Preliminary experiments indicated successful expression of ORCT, visualized through Western blotting. A complete understanding of the molecular structure, tissue expression, and physiological characterization of these organic cation transporters may hold promise for the formulation of more effective and environmentally benign insecticides, and may provide insights into the evolutionary origin of OCTs themselves.

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