Date of Award

Spring 2012

Document Type

Thesis

Degree Name

Bachelor of Science

Department

Biochemistry & Molecular Biol.

First Advisor

Dr. April Hill

Abstract

The evolution of gene regulatory networks accounts for much of the diversity we observe in the animal kingdom. In particular, the Pax/Six/Eya/Dac gene network has been known to play critical roles in development of all bilaterians and cnidarians. However, this network has not been characterized in Porifera. These studies focus on tracing the evolutionary history of the PSED network back to the most basal organism in the metazoan phylogeny through demonstrating the presence of a direct gene regulatory network between PaxB and Six1/2 orthologs in sponges. To this end, putatitve Six1/2 cis- regulatory elements were identified while the PaxBprd transcription factor was isolated and purified. Binding assays demonstrated that the PaxBprd protein bound directly to upstream cis-regulatory regions of the Six1/2 gene, as well as a conserved Six1/2 intron site in two different species of sponges. Additionally, we focused on broadening the Pax gene regulatory network in sponges by looking at additional downstream gene targets of Pax. We identified a possible indirect regulatory network between Pax and four novel

target genes through the use of RNAi and qRT-PCR techniques. This study helps to elucidate the origins of the PSED network as well as provide the beginnings of a whole

gene regulatory network in sponges.

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