Date of Submission

Spring 2013

Academic Program

Biology

Project Advisor 1

Brooke Jude

Abstract/Artist's Statement

Following the 2010 earthquake in Haiti, and after thousands of people were impoverished and displaced, cholera outbreak hit the country leaving more than eight thousands dead. Many studies have proposed various ways to fight this pandemic, which is caused by a new cholera strain, BAA-2163 El Tor variant.

Previous studies have shown that in addition to the toxin-coregulated pilus (TCP), there is another colonization factor, GlcNAc Binding Protein A (GbpA), that aids in the attachment of some wild-type El Tor V. cholerae strains to the chitinous exoskeletons of zooplanktons and the chitin monomers in small intestine of humans. The secretion of this protein is modulated by the HapR quorum sensing mechanism dependent on the bacterial density. There has been little research dedicated to studying this important colonization factor in the new Haitian El Tor variant. Our western blotting analyses demonstrated that the Haitian El Tor variant strain secretes GbpA and its level is dependent upon bacterial cell density, an indication that HapR quorum sensing is responsible for this fluctuation.

In order to explore its effect on the colonization of the Haitian El Tor variant, we attempted to construct BAA-2163 ΔgbpA strain by performing in-frame deletion. Initial experiments were unsuccessful in constructing the gbpA deletion plasmid via direct cloning. We propose that an efficient alternative technique could be TOPO cloning.

The HapR quorum sensing system along with GbpA facilitate the cholera pathogens transition between the aquatic and intestinal environments; thus, understanding their function in the Haitian variant could provide not only a new method for vaccine development, but also a novel way to limit its spread in aquatic environment in the long run.

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