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Characterisation and expression of ORF2 in genome segment 10 of African horse sickness virus
Dissertation (MSc (Genetics))--University of Pretoria, 2022.
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| Format: | Thesis |
| Language: | English |
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University of Pretoria
2023
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| _version_ | 1867613442958950400 |
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| access_status_str | Open Access |
| author2 | Van Staden, Vida |
| author_browse | Van Staden, Vida |
| author_facet | Van Staden, Vida |
| collection | Thesis |
| dc_rights_str_mv | © 2022 University of Pretoria. All rights reserved. The copyright in this work vests in the University of Pretoria. No part of this work may be reproduced or transmitted in any form or by any means, without the prior written permission of the University of Pretoria. |
| description | Dissertation (MSc (Genetics))--University of Pretoria, 2022. |
| format | Thesis |
| id | oai:repository.up.ac.za:2263/89251 |
| institution | University of Pretoria (South Africa) |
| language | English |
| last_indexed | 2026-06-10T12:36:13.446Z |
| license_str | Other — see source repository |
| provenance_str_mv | Harvested via OAI-PMH from UPSpace — University of Pretoria Institutional Repository |
| publishDate | 2023 |
| publishDateRange | 2023 |
| publishDateSort | 2023 |
| publisher | University of Pretoria |
| publisherStr | University of Pretoria |
| record_format | dspace |
| source_str | UPSpace — University of Pretoria Institutional Repository |
| spelling | oai:repository.up.ac.za:2263/89251 Characterisation and expression of ORF2 in genome segment 10 of African horse sickness virus Van Staden, Vida u17088900@tuks.co.za Potgieter, Christiaan Mizrachi, Eshchar Coertzen, Carmen African Horse Sickness Virus Additional open reading frames Synonymous constraint regions Segment 10 UCTD Dissertation (MSc (Genetics))--University of Pretoria, 2022. Viral genome segments are often under strong selective pressure to enhance the coding capacity of the genome, allowing expression of multiple proteins from a single mRNA. A second additional open reading frame, ORF2, was identified in African horse sickness virus (AHSV) Seg-10 which encodes the non-structural protein NS3. The ORF2 conservation, size, nature, expression, function and potential subcellular localisation of the putative protein product AHSV is not known and is investigated in this project. A bioinformatic analysis of all available AHSV Seg-10 sequence data was done. ORF2 was maintained in over 400 AHSV Seg-10 sequences, and six size variants were identified ranging from 183 to 252 nucleotides. ORF2 showed high conservation in the central region. AHSV Seg-10 ORF2 shows strong positive selection. The protein encoded by AHSV ORF2 was recombinantly expressed in insect and mammalian cells. Recombinant baculoviruses were confirmed to express ORF2 or ORF2-eGFP. Localisation of ORF2-eGFP was specific within the nucleus and cytoplasm of Sf9 cells. ORF2 transiently expressed in BSR-T7 cells initially localised in the cytoplasm and later moved to the nucleus, and ultimately caused cells to shrivel showing a cytotoxic effect. Antibodies raised against ORF2 was used to investigate the presence and subcellular localisation of the protein during AHSV infection. Using this antibody, ORF2 could be detected in AHSV infected cells, potentially identifying a novel AHSV non-structural protein. This is the first detection of a putative protein product of Seg-10 ORF2 during normal AHSV infection and paves the way for further functional analysis of this protein in the AHSV replication cycle. Poliomyelitis Research Foundation Deltamune Biochemistry, Genetics and Microbiology (BGM) MSc (Genetics) Unrestricted 2023-02-07T12:55:33Z 2023-02-07T12:55:33Z 2023-04 2022 Thesis * A2023 https://repository.up.ac.za/handle/2263/89251 10.25403/UPresearchdata.22005551 en © 2022 University of Pretoria. All rights reserved. The copyright in this work vests in the University of Pretoria. No part of this work may be reproduced or transmitted in any form or by any means, without the prior written permission of the University of Pretoria. application/pdf University of Pretoria |
| spellingShingle | African Horse Sickness Virus Additional open reading frames Synonymous constraint regions Segment 10 UCTD Characterisation and expression of ORF2 in genome segment 10 of African horse sickness virus |
| title | Characterisation and expression of ORF2 in genome segment 10 of African horse sickness virus |
| title_full | Characterisation and expression of ORF2 in genome segment 10 of African horse sickness virus |
| title_fullStr | Characterisation and expression of ORF2 in genome segment 10 of African horse sickness virus |
| title_full_unstemmed | Characterisation and expression of ORF2 in genome segment 10 of African horse sickness virus |
| title_short | Characterisation and expression of ORF2 in genome segment 10 of African horse sickness virus |
| title_sort | characterisation and expression of orf2 in genome segment 10 of african horse sickness virus |
| topic | African Horse Sickness Virus Additional open reading frames Synonymous constraint regions Segment 10 UCTD |
| url | https://repository.up.ac.za/handle/2263/89251 |