PURIFICATION AND CHARACTERIZATION OF ACETYLCHOLINESTERASE FROM DROSOPHILA SUZUKII

Date of Award

2013

Document Type

Thesis

Degree Name

Bachelors

Department

Natural Sciences

First Advisor

Walstrom, Katherine

Second Advisor

McCord, Elzie

Keywords

Drosophila, Acetylcholinesterase, Purification

Area of Concentration

Biochemistry

Abstract

Drosophila suzukii (Matsumura) is an invasive fly species and a current agricultural threat to soft peel fruit, such as strawberries and cherries, in the United States. Native to South East Asia, D. suzukii females have a serrated ovipositor that enables them to pierce the skin of ripening fruit to lay their eggs inside, which leads to the eventual decay of the fruit. D. suzukii has been exposed to pesticides throughout Asia, which may have conferred resistance via a mutated form of the enzyme acetylcholinesterase (AChE). AChE carries out a key step in the termination of synaptic transmission by hydrolyzing the neurotransmitter acetylcholine. AChE is the target of many current pesticides because its inhibition leads to ataxia, convulsions, and eventual death in the insects. In this study, AChE was purified from the heads of 5 day old D. suzukii flies by sequential extraction procedures followed by size exclusion chromatography. The partially purified AChE was used to optimize the pH and temperature of the reaction. Protein concentration assays were completed to calculate specific activity, and kinetic assays were performed. A Km value of 27 +/- 2 μM and a specific activity of 0.23 units/mg of protein were determined.

Rights

The author has not granted New College of Florida the nonexclusive right to archive, make accessible, and distribute for educational purposes this work in whole or in part in all forms of media, now or hereafter known. The copyright of this work remains with the author.

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