Dengue fever is a rapidly growing concern to human health and is currently the most prevalent mosquito-borne viral disease worldwide. Although there are several vaccine candidates being tested in clinical trials, there are no vaccines publicly available to prevent this disease. Plant-based vaccines are rapidly becoming viable alternatives to traditional animal-based vaccines because they are safe, easy to manufacture, and more cost-efficient. The purpose of this project is to develop a vaccine against the dengue virus by producing a recombinant DENV2 protein, engineered by Dr. David Lo and his lab at University of California Riverside, in Nicotiana benthamiana plants through Tobacco Mosaic Virus (TMV) infection. Initial attempts to ligate the complete DENV2 epitope, a combination of hybrid flagellin sequences and the envelope protein from dengue viral serotype 2, into the pJL TRBO vector were incompatible with established protocols. However, a proof of concept test that replaced the DENV2 envelope protein with Green Fluorescent Protein (GFP) successfully inserted the new sequence into pJL TRBO. In the future, the DENV2 envelope protein sequence will be re-inserted into the construct and updated protocols will be repeated for DENV2 protein expression. The recombinant DENV2 proteins will be extracted from the plants after signs of infection become apparent and tested for their ability to induce an immunogenic response that produces pathogen-specific antibodies.
| Identifer | oai:union.ndltd.org:CLAREMONT/oai:scholarship.claremont.edu:scripps_theses-1925 |
| Date | 01 January 2016 |
| Creators | Sheih, Tianna |
| Publisher | Scholarship @ Claremont |
| Source Sets | Claremont Colleges |
| Detected Language | English |
| Type | text |
| Format | application/pdf |
| Source | Scripps Senior Theses |
| Rights | © 2016 Tianna Sheih, default |
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