Construction of a Polyphosphate Kinase Gene Deletion in Yersenia spp.

Construction of a Polyphosphate Kinase Gene Deletion in Yersenia spp.

The goal of this experiment is to study the bacteria, Yersinia pestis, Yersenia enterocolitica, andYersenia pseudotuberculosis, and the possible role of the enzyme polyphosphate kinase (PPK) in pathogenicity, motility, and gene expression. PPK is an enzyme that produces inorganic polyphosphate, which is universal and found in all forms of life from prokaryotic bacteria to eukaryotic mammals. The PPK enzyme has been shown to be required for motility and virulence of several pathogenic bacteria.Yersinia spp. are diverse bacteria that cause diseases ranging from gastrointestinal diseases to bubonic plaque; however, the role of PPK is unknown. The focus will be on constructing the Y. pseudotuberculosis and Y. enterocolitica ppk deletion strains using λ Red-mediated gene replacement in order to understand the effects of deleting the ppk gene and determine if removal of the PPK enzyme affected motility and virulence. Determining a role for PPK in Yersinia pathogenicity could make it a prospective target for new antimicrobial drugs, innovative vaccines, and may have a great impact on the future.