Title
Characterization of Salmonella Typhimurium AB5 Enterotoxin
Document Type
Presentation
Publication Date
April 2010
Faculty Sponsor
Dr. Juliette Tinker
Abstract
Every year approximately 40,000 cases of Salmonella food poisoning are reported in the United States. While much is understood about Salmonella virulence, currently there is little known about the enterotoxins produced by Salmonella. Our goal with this study is to isolate and characterize the AB5 enterotoxins produced by this bacterium. After isolation and purification of this toxin we will be able to analyze the functional characteristics and compare it to similar AB5 toxins. With the evident need for novel vaccines this research is of importance in finding potential adjuvants that will stimulate a strong immune response with antigens associated with oral vaccines. We have screened and identified two strains of Salmonella Typhimurium that contain the correct gene sequence for the AB5 enterotoxin and cloned them into an appropriate vector for characterization. We have performed PCR to identify the presence of the ArtAB enterotoxin in two strains of Salmonella phagetype DT104. ArtAB was cloned into the pROExHTC plasmid to express and purify this toxin with an N-terminal Histine tag and a nickel column. In addition ArtAB was cloned into pTrchis-Topo to express and purify ArtAB with a C-terminal histine tag. After Isolation and purification of this toxin we will be able to analyze the functional characteristics and compare it to similar AB5 toxins. With the evident need for novel vaccines this research is of importance in finding potential new adjuvants that will stimulate a strong immune response with antigens administered as oral vaccines