Publication Date

Spring 2017

Degree Type

Thesis

Degree Name

Master of Science (MS)

Department

Chemistry

Advisor

Alberto A. Rascon, Jr.

Keywords

Aedes aegypti, bacteria, DNA, mosquito, proteases, recombinant expression

Subject Areas

Chemistry; Biochemistry; Molecular biology

Abstract

Aedes aegypti mosquitoes can be found globally in tropical and subtropical urban areas and spread Zika, Dengue fever, yellow fever, and Chikungunya viruses. Current vector control methods are limited and nonspecific. The female Ae. aegypti mosquito uses blood meal proteins to obtain nutrients required for oogenesis; inhibition of the midgut trypsin-like serine proteases responsible for blood meal digestion may provide a novel method of vector control. Ae. aegypti blood meal digestion is complex and the role of uncharacterized serine proteases in blood digestion is unclear; specifically, a group of trypsin-like serine proteases (AaSPII–V) is expressed at constant levels before and following Ae. aegypti blood meal acquisition. This research focuses on the in vitro biochemical study of two specific Ae. aegypti trypsin-like serine proteases (AaSPII and AaSPIV) in order to gain further understanding of their role in blood meal digestion. The approach involved the successful cloning and bacterial expression of these soluble, recombinant proteases. Results from attempts to purify these proteases were unsuccessful but indicative of potential autocatalytic and autodigestive behavior. Future studies will focus on obtaining purified recombinant proteases for further study. The study of AaSPII and AaSPIV, as well as other midgut Ae. aegypti proteases, will aid in understanding the overall role proteases play in blood meal digestion and may eventually allow for the development of mosquito-specific enzyme inhibitors.

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