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Genomic profile of eGFP-expressing canine distemper virus that undergoes serial plaque-to-plaque transfers
Canine distemper virus (CDV) is classified into the genus Morbillivirus in the family Paramyxoviridae. This virus has a single-stranded genomic RNA with negative polarity. The wild-type CDV genome is generally composed of 15 690 nucleotides. We previously rescued an enhanced green fluorescence prote...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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Frontiers Media S.A.
2022
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9545482/ https://www.ncbi.nlm.nih.gov/pubmed/36211954 http://dx.doi.org/10.3389/fcimb.2022.1006273 |
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author | Lin, Jiahui Jiang, Yujia Zhang, Hui Zhang, Feng Zhang, Youming Ni, Bo Liu, Fuxiao |
author_facet | Lin, Jiahui Jiang, Yujia Zhang, Hui Zhang, Feng Zhang, Youming Ni, Bo Liu, Fuxiao |
author_sort | Lin, Jiahui |
collection | PubMed |
description | Canine distemper virus (CDV) is classified into the genus Morbillivirus in the family Paramyxoviridae. This virus has a single-stranded genomic RNA with negative polarity. The wild-type CDV genome is generally composed of 15 690 nucleotides. We previously rescued an enhanced green fluorescence protein (eGFP)-tagged recombinant CDV (rCDV-eGFP) using reverse genetics. In this study, the rCDV-eGFP at passage-7 was subjected to 38 serial plaque-to-plaque transfers (or bottleneck passages) and two extra common passages in cells. In theory, the effect of Muller’s ratchet may fix deleterious mutations in a single viral population after consecutive plaque-to-plaque transfers. In order to uncover a mutated landscape of the rCDV-eGFP under the circumstances of bottleneck passages, the passage-47 progeny was collected for the in-depth analysis via next-generation sequencing. The result revealed a total of nine single-nucleotide mutations (SNMs) in the viral antigenome. Out of them, SNMs at nt 1832, 5022, 5536, 5580, 5746, 6913 and 8803 were identified as total single-nucleotide substitution, i.e., 100% of mutation frequency. The result suggested no notable formation of viral quasispecies in the rCDV-eGFP population after consecutive plaque-to-plaque transfers. |
format | Online Article Text |
id | pubmed-9545482 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
spelling | pubmed-95454822022-10-08 Genomic profile of eGFP-expressing canine distemper virus that undergoes serial plaque-to-plaque transfers Lin, Jiahui Jiang, Yujia Zhang, Hui Zhang, Feng Zhang, Youming Ni, Bo Liu, Fuxiao Front Cell Infect Microbiol Cellular and Infection Microbiology Canine distemper virus (CDV) is classified into the genus Morbillivirus in the family Paramyxoviridae. This virus has a single-stranded genomic RNA with negative polarity. The wild-type CDV genome is generally composed of 15 690 nucleotides. We previously rescued an enhanced green fluorescence protein (eGFP)-tagged recombinant CDV (rCDV-eGFP) using reverse genetics. In this study, the rCDV-eGFP at passage-7 was subjected to 38 serial plaque-to-plaque transfers (or bottleneck passages) and two extra common passages in cells. In theory, the effect of Muller’s ratchet may fix deleterious mutations in a single viral population after consecutive plaque-to-plaque transfers. In order to uncover a mutated landscape of the rCDV-eGFP under the circumstances of bottleneck passages, the passage-47 progeny was collected for the in-depth analysis via next-generation sequencing. The result revealed a total of nine single-nucleotide mutations (SNMs) in the viral antigenome. Out of them, SNMs at nt 1832, 5022, 5536, 5580, 5746, 6913 and 8803 were identified as total single-nucleotide substitution, i.e., 100% of mutation frequency. The result suggested no notable formation of viral quasispecies in the rCDV-eGFP population after consecutive plaque-to-plaque transfers. Frontiers Media S.A. 2022-09-23 /pmc/articles/PMC9545482/ /pubmed/36211954 http://dx.doi.org/10.3389/fcimb.2022.1006273 Text en Copyright © 2022 Lin, Jiang, Zhang, Zhang, Zhang, Ni and Liu https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
spellingShingle | Cellular and Infection Microbiology Lin, Jiahui Jiang, Yujia Zhang, Hui Zhang, Feng Zhang, Youming Ni, Bo Liu, Fuxiao Genomic profile of eGFP-expressing canine distemper virus that undergoes serial plaque-to-plaque transfers |
title | Genomic profile of eGFP-expressing canine distemper virus that undergoes serial plaque-to-plaque transfers |
title_full | Genomic profile of eGFP-expressing canine distemper virus that undergoes serial plaque-to-plaque transfers |
title_fullStr | Genomic profile of eGFP-expressing canine distemper virus that undergoes serial plaque-to-plaque transfers |
title_full_unstemmed | Genomic profile of eGFP-expressing canine distemper virus that undergoes serial plaque-to-plaque transfers |
title_short | Genomic profile of eGFP-expressing canine distemper virus that undergoes serial plaque-to-plaque transfers |
title_sort | genomic profile of egfp-expressing canine distemper virus that undergoes serial plaque-to-plaque transfers |
topic | Cellular and Infection Microbiology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9545482/ https://www.ncbi.nlm.nih.gov/pubmed/36211954 http://dx.doi.org/10.3389/fcimb.2022.1006273 |
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