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Patterson, Ian 
ORCID: https://orcid.org/0000-0003-3465-0848, Khanipov, Kamil, Swetnam, Daniele M., Walsdorf, Samantha, Kautz, Tiffany F., Thangamani, Saravanan, Fofanov, Yuriy and Forrester, Naomi L.
(2020)
'Measuring Alphavirus Fidelity Using Non-Infectious Virus Particles'. Viruses, Vol 12, Issue 5, e546.
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viruses-12-00546 - IPatterson May 20.pdf - Published Version Available under License Creative Commons Attribution. Download (820kB) | Preview |
Abstract
Mutations are incorporated into the genomes of RNA viruses at an optimal frequency and altering this precise frequency has been proposed as a strategy to create live-attenuated vaccines. However, determining the effect of specific mutations that alter fidelity has been difficult because of the rapid selection of the virus population during replication. By deleting residues of the structural polyprotein PE2 cleavage site, E3∆56-59, in Venezuelan equine encephalitis virus (VEEV) TC-83 vaccine strain, non-infectious virus particles were used to assess the effect of single mutations on mutation frequency without the interference of selection that results from multiple replication cycles. Next-generation sequencing analysis revealed a significantly lower frequency of transversion mutations and overall mutation frequency for the fidelity mutants compared to VEEV TC-83 E3∆56-59. We demonstrate that deletion of the PE2 cleavage site halts virus infection while making the virus particles available for downstream sequencing. The conservation of the site will allow the evaluation of suspected fidelity mutants across alphaviruses of medical importance.
| Item Type: | Article |
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| Additional Information: | This article belongs to the Special Issue Transmission Dynamics of Insect Viruses |
| Subjects: | QU Biochemistry > Genetics > QU 470 Genetic structures WC Communicable Diseases > Virus Diseases > General RNA Virus Infections > WC 501 RNA virus infections (General or not elsewhere classified) WC Communicable Diseases > Virus Diseases > Infectious Mononucleosis. Arbovirus Infections > WC 524 Arbovirus infections |
| Faculty: Department: | Biological Sciences > Vector Biology Department |
| Digital Object Identifer (DOI): | https://doi.org/10.3390/v12050546 |
| Depositing User: | Mel Finley |
| Date Deposited: | 21 May 2020 14:20 |
| Last Modified: | 21 May 2020 14:20 |
| URI: | https://archive.lstmed.ac.uk/id/eprint/14483 |
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