Trypanosoma cruzi recombinant protein expression in Escherichia coli
Document Type
Oral Presentation
Campus where you would like to present
SURC Ballroom C/D
Start Date
15-5-2014
End Date
15-5-2014
Keywords
Chagas Disease, Paraflagellar Rod, Vaccine Potential
Abstract
Chagas disease, or American Trypanosomiasis, is a parasitic disease found throughout Central and South America. It is caused by the single-celled parasite, Trypanosoma cruzi, and transmitted by Triatominae, or kissing bug, a large blood-sucking insect that is often found in rural adobe homes. Trypanosomes have a unique structure, the paraflagellar rod (PFR), which runs along the length of the flagellum. The PFR is composed of a lattice of cytoskeletal filaments and is critical for cell motility. The proteins of the PFR in T. cruzi have been shown to be immunogenic, protecting mice from an otherwise lethal challenge with the parasite. Two previously unidentified PFR-like genes, PFR-5 and PFR-6, were discovered when the T. cruzi genome was sequenced. The aim of this project was to determine if these two putative PFR proteins are associated with the flagellum. Portions of the PFR-5 and PFR-6 genes have been cloned into expression plasmids. These plasmids are expressed in Escherichia coli to produce recombinant proteins which would be harvested, purified, and injected into mice to generate PFR-specific antibodies. Unfortunately, no recombinant protein was detected in the E. coli cultures. Further research will require the recloning of the PFR-genes to obtain recombinant E. coli.
Recommended Citation
McDonald, Jay, "Trypanosoma cruzi recombinant protein expression in Escherichia coli " (2014). Symposium Of University Research and Creative Expression (SOURCE). 33.
https://digitalcommons.cwu.edu/source/2014/posters/33
Poster Number
24
Additional Mentoring Department
Biological Sciences
Trypanosoma cruzi recombinant protein expression in Escherichia coli
SURC Ballroom C/D
Chagas disease, or American Trypanosomiasis, is a parasitic disease found throughout Central and South America. It is caused by the single-celled parasite, Trypanosoma cruzi, and transmitted by Triatominae, or kissing bug, a large blood-sucking insect that is often found in rural adobe homes. Trypanosomes have a unique structure, the paraflagellar rod (PFR), which runs along the length of the flagellum. The PFR is composed of a lattice of cytoskeletal filaments and is critical for cell motility. The proteins of the PFR in T. cruzi have been shown to be immunogenic, protecting mice from an otherwise lethal challenge with the parasite. Two previously unidentified PFR-like genes, PFR-5 and PFR-6, were discovered when the T. cruzi genome was sequenced. The aim of this project was to determine if these two putative PFR proteins are associated with the flagellum. Portions of the PFR-5 and PFR-6 genes have been cloned into expression plasmids. These plasmids are expressed in Escherichia coli to produce recombinant proteins which would be harvested, purified, and injected into mice to generate PFR-specific antibodies. Unfortunately, no recombinant protein was detected in the E. coli cultures. Further research will require the recloning of the PFR-genes to obtain recombinant E. coli.
Faculty Mentor(s)
Stryker, Gabrielle