Poliomos virusų paieška, genomų sekų nustatymas ir analizė Europoje sugautų graužikų mėginiuose

• Main Author: Vasiliūnaitė, Emilija
• Other Authors: Gedvilaitė, Alma
• Format: Master Thesis
• Language: Lithuanian; English
• Published: Institutional Repository of Vilnius University 2020
• Subjects: Microtus arvalis; Rattus rattus
• Online Access: http://vu.oai.elaba.lt/documents/69466526.pdf; http://vu.lvb.lt/VU:ELABAETD69466526&prefLang=en_US

Search, Identification and Analysis of Polyomavirus Genome Sequences in Rodent Samples From Europe Polyomaviruses (PyVs) are ~ 5 kb double-stranded DNA viruses that infect mammals, birds, and fish. Recently, pathogenesis and evolution of PyVs are extensively studied. Rodents, the largest group of mammals, have been relatively little investigated for PyVs, making them a good target for searching for new polyomaviruses. Rodents and their PyVs are a suitable model for the analysis of PyV host-virus coevolution. In this study, PyV-specific hexanucleotide primers were developed for the strand displacement amplification reaction. However, novel primers did not amplify PyV DNA significantly more efficiently and did not significantly increase the ratio of PyV DNA to mitochondrial DNA in samples compared to random primers using Phi29 and EquiPhi29 polymerases for amplification. The rodent samples of genera Microtus, Apodemus, and Rattus were examined for polyomaviruses. One Microtus arvalis vole had a PyV sequence, 99% identical to CVPyV. A new polyomavirus, MarPyV2, was identified in 14 Microtus arvalis voles and had a genomic sequence identity of 73% to MPtV and 62% to CVPyV. Co-evolution with the host and loss were the most important events for the inter-population diversity of this virus. AflaPyV1 DNA was detected in one Apodemus flavicollis mouse, and no PyV DNA was detected in Apodemus agrarius mice. In Rattus rattus rats, a new PyV with 92% genomic sequence identity to RnorPyV1 and a variant of RnorPyV1 with a 1226 bp deletion spanning the NCCR and adjacent sequences were identified. The synthesis of RnorPyV1 VP1 protein in yeast cells resulted in a yield of 0.389 mg per 1 g of wet yeast biomass, and the protein formed virus-like particles of 43 ± 5.3 nm in size. RatPyV2 VP1 protein synthesis was not noticeable in yeast cells.