Rotavirus (RV) is a pathogen that causes gastroenteritis in humans and animals. Although there are A-G types of rotavirus, type A (RVA) is the most commonly studied because it is the leading cause of death in children under 5 years averaged with 200,000 deaths a year. Understanding the infection biology of group B rotavirus (RVB) is essential because it has the potential to cause outbreaks in adults. Because RVB is not heavily studied, there are not many protocols for the culture techniques which is one aim of this research. Until 2021, equine rotaviruses were thought to only be of RVA, but equine rotavirus B (EVRB) has recently been associated with neonatal foal diarrhea outbreaks. This research will focus on both foal and equine Rotavirus B and how the VP8* protein interacts with the host. Through work with X-Ray Crystallography, the static structure of the VP8* protein will be obtained. An NMR experiment will be performed in order to determine the dynamic structure of the protein. One hypothesis of this research is that the dynamic structure of VP8* is involved in the binding, meaning the minority population needs to be analyzed. The results of the sample preparation and gel analysis showed that the arctic expression vector was the most promising option. An equine arctic RVB VP8* NMR sample was able to be prepared and an HSQC spectrum confirmed our protein. It was determined that the RVB VP8* protein is suitable for an NMR experiment which allows for subsequent testing. Future work will include a binding study as well as more buffer optimization experiments because the two buffers used did not result in a high protein concentration. Overall, this research will allow for a crucial step in developing culture techniques and understanding infection biology of RVB.
Author(s): Maggie Blossey, Shuisong Ni, Michael A. Kennedy, PhD.
Advisor(s): Michael Kennedy, Department of Chemistry and Biochemistry
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