CLONING A VIRULENCE GENE FROM BORDETELLA AVIUM

Molly Baum, Alice Chiang, Joshua Cook, Christina Crum, Karina Lam, Meghana Limaye, Varun Ponmudi, Susan Putnins, Thomas Riemschneider, Mirat Shah, Monica Thanawala

Advisor: Dr. Louise Temple
Assistant: Karen Riesenburger
Consultants: Jen Brady, Angella Dorsey-Oresto, Gene Gillespie,
Mary Gomes, Matt Hui, Kelly Prescott, Dr. Munju Tyagi

Bordetella avium (B. avium) causes bordetellosis, a disease similar to whooping cough, in turkeys and other birds. B. avium has proteins on its outer membrane called fimbriae which attach to the cilia in the turkey’s trachea. The aim of this project was to isolate the fimbrial genes in order to synthesize fimbrial proteins and to eventually create a component vaccine. Polymerase chain reaction was performed on four different B. avium genes expected to code for fimbriae: fimA, fim1, fim2, and fim3. Three genes were successfully amplified, and were subsequently ligated into the cloning vector, pET151/D- TOPO. The ligated plasmids were then transformed into Escherichia coli (E. coli) to allow the plasmids to replicate. The plasmids were isolated and examined, and although electrophoresis did not prove that ligation had occurred, the E. coli were cultivated to synthesize the fimbrial protein from the plasmid DNA. Protein gel analysis showed that these cells did not contain proteins of the expected size, which leads to the conclusion that the fimbriae genes were not properly expressed. Future research should continue attempts to synthesize the fimbriae protein because this is an essential step to developing a working component vaccine.

Paper

(Acrobat pdf file: 920 kb)

Presentation

(Acrobat pdf file: 1.3 Mb)

Team Project Picture (jpg: 301 kb)

Return to Team Projects