Date of Award

5-2023

Document Type

Thesis

Degree Name

Master of Science (MS)

Department

Biological Sciences

Committee Chair/Advisor

Matthew Turnbull

Committee Member

David Feliciano

Committee Member

Min Cao

Abstract

Testing gene expression patterns is an important process in learning more about an organism. The standard methods of rtPCR and RNASeq provide highly detailed data on specific expression patterns, but can be resource consuming if many different samples including genes or organisms must be tested. A potential alternative for studying expression is the use of a reporter system carried by a vector system that encodes fluorescent proteins under the control of promoters of interest. Many lepidopterans (moths) are pestiferous and better understanding of gene expression levels in lepidopterans, both endogenous and exogenous including from their viruses, would be beneficial. Baculoviruses have been used widely as vectors to deliver genetic elements to lepidopteran hosts. Here, I developed a first generation baculovirus reporter vector to analyze gene expression regulatory elements in lepidopterans. I generated backbone transfer plasmids and viruses, including recombinant experimental viruses incorporating previously examined polydnavirus promoter elements. Recombinant viruses were used to infect Sf9 insect cells and fluorescence emission was measured by plate reader at several multiplicities of infection and times post-infection to estimate promoter activity. Infected cells were also observed via microscopy. I observed repeatable emission patterns associated with the inserted promoter elements, while identifying possible issues associated with the chosen reporter system that may be addressed in future modifications.

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