Senior Honors Projects, 2010-current

Date of Award

Spring 2017

Document Type

Thesis

Degree Name

Bachelor of Science (BS)

Department

Department of Integrated Science and Technology

Advisor(s)

Louise Temple, Ph.D.

Stephanie Stockwell, Ph.D.

Christopher Berndsen, Ph.D.

Abstract

A Gram negative species of bacteria, Campylobacter jejuni, is the leading cause of food poisoning worldwide. Humans often contract food poisoning after ingesting contaminated poultry. Detecting the presence of C. jejuni in poultry is difficult because it is part of the natural flora and does not cause symptomatic infection. In a related manner, Bordetella avium is a Gram negative species of bacteria that causes bordetellosis in poultry. This disease is similar to whooping cough caused by the related pathogen of humans, B. pertussis. Though the mortality rate for bordetellosis is low, it weakens the birds’ immune systems, often leading to secondary infections. The aim of this project was to construct a vaccine platform capable of immunizing poultry against both pathogens—B. avium and C. jejuni—thus reducing disease in birds and humans. A heterologous construct can be made utilizing the B. avium autotransporter Baa1 that plays a role in host cell attachment. Autotransporters are comprised of three genetic regions: promoters to drive expression, and encoded passenger and transporter domains. The transporter is a beta barrel anchored in the outer membrane. The passenger domain is translocated through the transporter and secreted to the outer surface of the bacterial cell. An antigenic Campylobacter gene was cloned into the encoded passenger domain of the Baa1 autotransporter in the context of a suicide plasmid. Tri-parental mating was done to promote homologous recombination of the construct into the B. avium chromosome. After concluding the construct was not in the chromosome most likely due to instability, a plasmid was synthesized and the codons of the Campylobacter region were optimized. Tri-parental mating was completed again. All steps were verified with PCR and gel electrophoresis. A B. avium transconjugant containing the chimeric cja::baa1construct was isolated. Future work will involve gene expression and challenge studies.

Included in

Biotechnology Commons

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