Cloning and Purification of the ArtB Binding Subunit From a Novel Toxin in Salmonella enterica Typhimurium
Dr. Juliette Tinker
According to the CDC, Salmonella is responsible for about 1.35 million infections, 26,500 hospitalizations, and 420 deaths in the United States every year. Food is the source for most of these illnesses. Salmonella enterica is common in cattle and has a number of serovars that are pathogenic in humans. Many Salmonella serovars encode for the AB5 toxin, or ArtAB, which is similar in structure to pertussis toxin. The pentameric B subunit is non-covalently linked to the A subunit, thus allowing separation of the receptor-binding B subunit from the holotoxin. We hypothesize that being able to extract and use ArtAB or the non-toxic ArtB subunit for vaccines could decrease prevalence of Salmonella infections in cows and potentially prevent transmission from cattle and agriculture to humans. The B subunit can be transformed into E. coli using a vector plasmid and then purified and extracted using D-galactose and fetuin affinity chromatography methods. Our results show that we have successfully cloned the B subunit into a plasmid, transformed the plasmid into E. coli, and had ArtB protein expression. Further research will examine the toxicity of ArtAB and ArtB to determine whether or not they are suitable for use in vaccines.
Cufurovic, Leyla; Rodriguez, Adriana; Mousa, Omid Mohammad; and Tinker, Juliette, "Cloning and Purification of the ArtB Binding Subunit From a Novel Toxin in Salmonella enterica Typhimurium" (2020). 2020 Undergraduate Research Showcase. 37.