CONSTRUCTION AND CHARACTERIZATION OF REPORTER STRAINS FOR THE IDENTIFICATION OF PHYSIOCHEMICAL PARAMETERS INFLUENCING...OF ESCHERICHIA COLI

Identify appropriate primers and PCR amplify the promoters and upstream regions for eaeA, stx2, hns, and rpoS. Ligate the amplified promoters into a promoter-less fluorescent protein vector. Transform into E. coli O157:H7 and compare the survival with the plasmid-free parent strain. Monitor the promoter-EFP expression during growth and following exposure to various physiochemical parameters. Conduct studies on ground beef inoculated with the engineered strains to determine the utility of these strains in identifying food-related parameters that influence the expression of these virulence and regulatory genes.