Publication Date

Spring 2017

Degree Type

Thesis

Degree Name

Master of Science (MS)

Department

Chemistry

Advisor

Alberto A. Rascón

Keywords

AaSPVII, Aedes aegypti, blood meal, dengue, midgut serine protease, Zika

Subject Areas

Biochemistry

Abstract

The Aedes aegpyti mosquito is an efficient biological vector of four known neglected tropical arboviruses: the yellow fever, dengue, chikungunya, and Zika viruses. Since the female mosquito can acquire multiple blood meals to complete the gonotrophic cycle, she can potentially infect multiple hosts. With the reemergence of these viruses, especially due to global warming, it is crucial to slow the spread of the mosquito and the blood-borne diseases she transmits. The Ae. aegypti mosquito relies heavily on midgut serine proteases that digest blood meal proteins to provide nutrients required for the egg laying process. The focus of this work is on a late phase midgut serine protease, AaSPVII. AaSPVII is of interest due to its potential difference in specificity and functional role in the digestion process, despite its sequence similarity to another late phase protease known as AaSPVI. By understanding the important role these midgut serine proteases play, we could potentially inhibit these proteases and disrupt fecundity. Currently, two different expression constructs were produced, one with an N-terminal His6-tag and the other with a C-terminal His6-tag. Both constructs have been recombinantly expressed using bacteria, but only the C-terminally His6-tagged AaSPVII has been purified and activated successfully. This enzyme has comparable steady-state kinetic parameters to previously published results, suggesting that we have optimized soluble bacterial expression.

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