Cargando…

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...

Descripción completa

Detalles Bibliográficos
Autores principales: Lin, Jiahui, Jiang, Yujia, Zhang, Hui, Zhang, Feng, Zhang, Youming, Ni, Bo, Liu, Fuxiao
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2022
Materias:
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
_version_ 1784804829950902272
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
work_keys_str_mv AT linjiahui genomicprofileofegfpexpressingcaninedistempervirusthatundergoesserialplaquetoplaquetransfers
AT jiangyujia genomicprofileofegfpexpressingcaninedistempervirusthatundergoesserialplaquetoplaquetransfers
AT zhanghui genomicprofileofegfpexpressingcaninedistempervirusthatundergoesserialplaquetoplaquetransfers
AT zhangfeng genomicprofileofegfpexpressingcaninedistempervirusthatundergoesserialplaquetoplaquetransfers
AT zhangyouming genomicprofileofegfpexpressingcaninedistempervirusthatundergoesserialplaquetoplaquetransfers
AT nibo genomicprofileofegfpexpressingcaninedistempervirusthatundergoesserialplaquetoplaquetransfers
AT liufuxiao genomicprofileofegfpexpressingcaninedistempervirusthatundergoesserialplaquetoplaquetransfers