The study of viral capsid proteins, such as VP2 of CtenDNAV-II, is essential for understanding viral capsid assembly, infection mechanisms, and host manipulation. Capsid proteins, the protein shells encapsulating viral genetic material, play critical roles in protecting the virus and aiding in its attachment to host cells. By capturing high-resolution images of individual virus particles embedded in vitrified ice, cryo-EM facilitates the reconstruction of three-dimensional (3D) structures of viral capsids. The study of viral capsid structures using cryo-EM provides insights into viral life cycles, host-virus interactions, and evolutionary events. The aim of this research was to gain deeper insights into viral capsid architecture and dynamics by reconstructing the small capsid particle (T=1) and investigating its role in the capsid assembly of CtenDNAV-II. The capsid protein was expressed using Sf9 insect cells that were inoculated with a baculovirus containing the VP2 gene. The capsid protein expression in E. coli was performed by transforming the cells with a ubiquitin-expressing plasmid. An already existing cryo-EM dataset, which has been used to reconstruct the large capsid particle (T=3) was used to reconstruct the small capsid particle and determine whether or not it was hollow inside. The single particle reconstruction of the T=1 capsid particles, realized using cryoSPARC, achieved a fairly high resolution and indicated that the particles are most likely empty inside, containing no genome. It was noticed that many of the small particles were broken, which indicates they were most likely intermediate or incomplete VLP assembly states.
| Identifer | oai:union.ndltd.org:UPSALLA1/oai:DiVA.org:his-24152 |
| Date | January 2024 |
| Creators | Stoe, Armand |
| Publisher | Högskolan i Skövde, Institutionen för biovetenskap |
| Source Sets | DiVA Archive at Upsalla University |
| Language | English |
| Detected Language | English |
| Type | Student thesis, info:eu-repo/semantics/bachelorThesis, text |
| Format | application/pdf |
| Rights | info:eu-repo/semantics/openAccess |
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