Generation of Fluorescent Recombinant Listeriolysin O Toxin for Analysis of Interactions with Host Protein
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Date
2008-06
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The Ohio State University
Abstract
Listeriolysin O is the major virulence factor of Listeria monocytogenes and is a member of the Cholesterol-Dependant Cytolysin (CDC) family of pore-forming toxins. The CDCs are a crucial virulence factor secreted by Gram-positive bacteria, however the mechanisms underlying host cell subversion by this family of toxins are poorly understood. The goal of this project is to obtain recombinant Listeriolysin O which, when fluorescently labeled, can be used to determine the spatial and temporal dynamics of LLO interactions with host cells. In order to achieve this goal, the project includes two major steps: 1) construction, purification, and fluorescent labeling of the recombinant toxin and 2) analysis of the interplay between toxin and host cells by fluorescence microscopy. Using LLO as a model CDC, this study will lay the groundwork for understanding how CDCs hijack host cells. Determining how CDCs function will bring a better understanding of diseases generated by Gram-positive bacteria. Also, the results will allow us to refine the design of antitoxins to fight infectious diseases.
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Keywords
Listeriolysin O