Transient expression of the rabies glycoprotein in soybean

WCU Author/Contributor (non-WCU co-authors, if there are any, appear on document)
Grayson Williams (Creator)
Institution
Western Carolina University (WCU )
Web Site: http://library.wcu.edu/
Advisor
Amanda Storm

Abstract: Rabies is a highly preventable disease that is still claiming thousands of lives every year. An edible rabies vaccine for animals would reduce the cost and resources needed for the vaccine to reach less developed regions by simplifying administration of the vaccine and removing the need for cold-chain transport. When expressed on its own, the glycoprotein of rabies has been shown to form virus-like particles (VLPs), which structurally resemble the native virus without any chance of being infectious. VLPs of other viruses produced in plants have been shown to produce varying levels of immunity in animal studies, and some virus VLPs are used today in FDA approved vaccines. The promise of VLP vaccines coupled with soybean as a promising system due to its scalable production, protein rich seeds, and extensive leaf material, makes an edible vaccine possible. This research presents the optimization of a successful agroinfiltration method for soybean leaf tissue, as well as production of a plasmid containing a GUS (ß glucuronidase) reporter gene and the rabies Glycoprotein (G protein) containing a His-tag and ER (endoplasmic reticulum) retention signal sequences for future work towards the transient and transgenic expression of the G protein in soybean. A syringe-mediated injection agroinfiltration method for effective transient transformation of soybean leaf tissue has been developed. The new plasmid will allow for simple detection of transformed soybean tissue using GUS, and isolation and detection of the G protein in the transformed samples using the His-tag. The findings of this project provide protocols and products developed to work toward a successful edible rabies vaccine for animals. While this research lays the foundation for producing an edible vaccine for animals, this is a steppingstone toward a much greater goal. Edible vaccines for humans would overcome the limitations that hold traditional vaccines back from reaching many underdeveloped regions, where people are still dying from preventable disease.

Additional Information

Publication
Thesis
Language: English
Date: 2021
Subjects
Rabies virus
Glycoproteins
Vaccines
Soybean

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