Title

Trypanosoma cruzi recombinant protein expression in Escherichia coli

Presenter Information

Jay McDonald

Document Type

Oral Presentation

Location

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.

Poster Number

24

Faculty Mentor(s)

Stryker, Gabrielle

Additional Mentoring Department

Biological Sciences

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May 15th, 8:30 AM May 15th, 11:00 AM

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.