Assessment of the Role of Brucella abortus Type-V Auto-secreted Proteins in the Pathogenesis and Host Immunity of Brucellosis
Hornay, Alyssa
Hornay, Alyssa
Abstract
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Brucella abortus is the etiological agent of brucellosis in cattle and elk and is known to cause abortion, sterility and decreased milk production. Two vaccines have previously been developed for brucellosis. Immunization with strain 19, the first vaccine to be developed and used, is highly reactogenic and not recommended for use in wildlife. The current vaccine strain, RB51, is able to induce protection from natural brucellosis infection in cattle but not in wildlife. This study was designed to evaluate a protein from B. abortus that may potentially make an excellent candidate for induction of protective immunity and a possible subunit vaccine candidate. The gene product encoded by AidA, an auto-secreted putative adhesin, has been found to be up-regulated during infection, using a gene discovery method known as In-vivo Induced Antigen Technology (IVIAT). Presently, AidA was amplified by polymerase chain reaction (PCR) and successfully transformed into competent Escherichia coli cells using the pETBlue AccepTorVector. Induced expression of AidA was analyzed by SDS-PAGE and Western blotting. Preliminary results indicated that the recombinant protein levels were nominal and optimization methods to stably express the gene are currently being developed. Once produced, if recombinant AidA is better able to induce protective immunity, it may prove to be safer and more reliable than the current live attenuated platforms.
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University of Wyoming Libraries