Publication Date

Summer 2018

Degree Type

Thesis

Degree Name

Master of Science (MS)

Department

Chemistry

Advisor

Alberto A. Rascon, Jr.

Keywords

Autocatalytic, Mosquito, Protease, Recombinant Expression, Serine Protease, Vector Control

Subject Areas

Biochemistry

Abstract

The female Aedes aegypti mosquito requires specific midgut proteases to digest

blood meals in a biphasic manner for completion of the gonotrophic cycle. The anautogenous nature of the female mosquito has led to the mosquito in becoming an efficient vector in transmitting several bloodborne pathogens: Chikungunya, Yellow Fever, Dengue and the Zika virus. With the many flaws associated with current vector control strategies, our research lab is focused on the development of a potential novel vector control strategy by targeting midgut proteases involved in blood meal digestion. The role of one of the four most abundantly expressed proteases, AaET, an early phase trypsin-like serine protease, is the focus of investigation. Characterization of AaET is needed to understand the overall relationship between the mosquito midgut digestion and viral transmission. N-terminal and C-terminal His6-tagged AaET, AaET with a pseudo propeptide (EK), and two mutant AaET genes were cloned and recombinantly expressed in bacterial cells. N-terminal His6-tagged AaET and AaET-EK were purified and tested for activity in vitro. The inactive AaET (WT) zymogen potentially exhibits autocatalytic behavior contributing to purification challenges and low protein yields.

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