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Construction of a novel bat coronavirus glycoprotein pseudotyped lentiviral vector and analysis of cell tropism

Haynes, Soraya (2019) Construction of a novel bat coronavirus glycoprotein pseudotyped lentiviral vector and analysis of cell tropism. Honours thesis, Murdoch University.

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Since the SARS epidemic in 2003, and the subsequent identification of bats as the reservoir host, there has been a surge of interest in research into bat-borne viruses, with over 30 new bat coronaviruses alone being discovered. Investigating the cell tropism of these newly discovered coronaviruses deepens the understanding of their pathology and possible host ranges, as well as allowing assessments to be made on their zoonotic potential. Aside from biosafety concerns however, this is also reliant on isolation of the virus from its host, which can be difficult using conventional methods.

This project aimed to develop a second generation lentivirus pseudotyping system which would allow spike proteins, the main determinants of coronavirus cell entry, of a non-isolated coronavirus to be safely expressed on a lentiviral particle in order to assess its cell tropism. The system was developed and tested using the spike gene from CoV1087, a novel bat-borne coronavirus with a 70% similarity at both the coding RNA and amino acid level between its spike protein and that of porcine epidemic diarrhea virus strain CH-HNYF-14.

Overall this study resulted in the successful development of a lentiviral pseudotyping system which allows the efficient analysis of the cellular tropism of coronaviruses. Through the testing and development of this system, it was determined that CoV1087 could enter two human cell lines, 293T and CaCo-2 as well as one rabbit cell line, RK-13. This raises the possibility of the transfer of the virus to feral rabbits as well as the possibility of zoonotic human infections.

Item Type: Thesis (Honours)
Murdoch Affiliation(s): School of Veterinary and Life Sciences
Supervisor(s): O'Dea, Mark, Abraham, Sam and Muhling, Jillian
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