dsRNA Infection of Protozoan, Euglena
Additional Funding Sources
The project described was supported by the Research Experience for Undergraduates Program Site: Molecular and organismal evolution at the University of Idaho under Award No. 1757826.
Presentation Date
7-2019
Abstract
In recent years, the presence of double-stranded RNA viruses have been linked to increased pathogenicity of Protozoa, including the human parasite, Leishmania, the causative agent of Leishmaniasis. Also known as Black Fever, the disease causes ~70,000 deaths each year. Virally infected pathogens such as these share the phylum Euglenozoa with the non-virulent Euglena. The potential presence and composition of viruses in various Euglenid species could illuminate differences in capacity for viral infection between harmless Euglena species and their pathogenic relatives. To address this, dsRNA was extracted and purified using an elegant approach that exploits affinity chromatography. To begin, Euglena cultures were harvested and pulverized using mortar and pestle with liquid nitrogen. Proteins and lipids were then removed from the concentrated Euglena cells using phenol-chloroform and SDS extraction. Samples were then filtered using cellulose columns that bind dsRNA. The dsRNA eluate was analyzed using gel electrophoresis. After imaging, viral presence may be denoted by dsRNAs of approximately 3.5 and 2.2 kb. These sizes correspond to the family of Tombus-like viruses commonly found in Euglena’s Leishmania relatives. The experiments of this study are still ongoing. Pending the presence of dsRNA viruses in Euglena, future work may focus on characterization of these viruses.
dsRNA Infection of Protozoan, Euglena
In recent years, the presence of double-stranded RNA viruses have been linked to increased pathogenicity of Protozoa, including the human parasite, Leishmania, the causative agent of Leishmaniasis. Also known as Black Fever, the disease causes ~70,000 deaths each year. Virally infected pathogens such as these share the phylum Euglenozoa with the non-virulent Euglena. The potential presence and composition of viruses in various Euglenid species could illuminate differences in capacity for viral infection between harmless Euglena species and their pathogenic relatives. To address this, dsRNA was extracted and purified using an elegant approach that exploits affinity chromatography. To begin, Euglena cultures were harvested and pulverized using mortar and pestle with liquid nitrogen. Proteins and lipids were then removed from the concentrated Euglena cells using phenol-chloroform and SDS extraction. Samples were then filtered using cellulose columns that bind dsRNA. The dsRNA eluate was analyzed using gel electrophoresis. After imaging, viral presence may be denoted by dsRNAs of approximately 3.5 and 2.2 kb. These sizes correspond to the family of Tombus-like viruses commonly found in Euglena’s Leishmania relatives. The experiments of this study are still ongoing. Pending the presence of dsRNA viruses in Euglena, future work may focus on characterization of these viruses.
Comments
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