Date of Submission

Spring 2013

Academic Program

Biology

Project Advisor 1

John Ferguson

Project Advisor 2

Brooke Jude

Abstract/Artist's Statement

Abstract

The Haitian cholera epidemic of 2010 has led to increased interest in virulence factors of Vibrio cholerae, the bacterial agent responsible for cholera. The ATP-yielding Entner-Doudoroff (ED) pathway is widely distributed in gram-negative bacteria, including V. cholerae, and fulfills vital roles in central metabolism. One key enzyme that is distinctive and highly important to the ED pathway is 6-phosphogluconate dehydratase (EDD). In this study, evidence of the ED pathway in a quorum sensing positive strain of V. cholerae (El Tor strain C6706) was obtained through growth on gluconate supplemented agar plates. This research began the construction of a deletion plasmid, which aims to silence the edd gene and deactivate the ED pathway in V. cholerae C6706. Previous research on the ED pathway in a quorum sensing negative strain of V. cholerae revealed that the ED pathway upregulated prime virulence genes ctxA and tcpA and downregulated biofilm formation. Quorum sensing controls biofilm formation in V. cholerae C6706 through cell-to-cell communication, making the presence or absence of biofilms directly related to cell density. Based on previous research on ED pathway significance, I hypothesize that upon the creation of an edd C6706 mutant strain, cholera toxin production would decrease. However, due to the fact El Tor C6706 is quorum sensing positive, I hypothesize that biofilm formation will increase by an insignificant amount in the edd mutant. The study of the ED pathway in V. cholerae indicates its considerable influence in survival and pathogenesis.

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