Date of Award

Spring 5-8-2017

Document Type

Thesis

Degree Name

Bachelor of Arts

Department

Biochemistry & Molecular Biology

First Advisor

Chris Gillen

Abstract

Aedes aegypti (Yellow Fever Mosquito) is a vector for many diseases, including Zika and yellow fever. With the increase in mosquito resistance to insecticides, new molecular targets for pesticides are needed. One potential protein target are the sodium dependent cation-chloride cotransporters (CCCs) which may play a role in salt and water balance in both larval and adult life stages of A. aegypti. In this study, we measured the protein expression of aeCCC2 and aeCCC3 using western blots. Polyclonal antibodies were developed by injecting two rabbits with two peptide sequences from each paralog. Antiserum from both rabbits reacted with proteins in larvae, adult male, and adult female whole body tissues with the most prominent bands above the 150 kDa size marker on 7.5% SDS-PAGE gels. To develop paralog-specific antibodies, the antiserum was affinity purified against peptides from either aeCCC2 or aeCCC3. We confirmed specificity of these antibodies with peptide competition experiments on dot blots. Similar to the unpurified serum, the affinity purified antibodies reacted with proteins above 150 kDa on western blots of whole body lysates and membrane preps of larvae and adults, both males and females. When testing the protein expression between different life stages, we found that antiserum purified against aeCCC2 peptides had on average an 18-fold higher reactivity in adults than in larvae and that antiserum purified against aeCCC3 had an 22-fold higher reactivity in larvae than in adults.

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