Purification of Novel Shiga Toxin Based Vaccine Adjuvants
Dr. Juliette Tinker
Shiga toxin is a bacterial enterotoxin that represents a promising vaccine adjuvant. The purpose of this experiment is to purify a detoxified derivative of Shiga toxin. These experiments have focused on cloning the StxB gene, a subunit of Shiga toxin, into the commercially available plasmid pBAD18. The StxB gene will be cloned to create three plasmids. The first plasmid will contain StxB with a 6x Histidine tag to allow for nickel column purification. This plasmid will also have a multiple cloning site to allow for the addition of antigens to the subunit protein. The second plasmid will have just the StxB gene alone. The purpose of this plasmid is to test if the StxB subunit can be purified using D-galactose agarose. The final plasmid will contain the StxB subunit and a multiple cloning site. This plasmid can be used to test if StxB can be purified with D-galactose while also bound to an antigen. Once the plasmids have been constructed, further experiments will be conducted to determine the best method of purification for Shiga toxin derived vaccine adjuvants.
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