Cargando…

Recovery of genetically defined murine norovirus in tissue culture by using a fowlpox virus expressing T7 RNA polymerase

Despite the significant disease burden caused by human norovirus infection, an efficient tissue-culture system for these viruses remains elusive. Murine norovirus (MNV) is an ideal surrogate for the study of norovirus biology, as the virus replicates efficiently in tissue culture and a low-cost anim...

Descripción completa

Detalles Bibliográficos
Autores principales: Chaudhry, Yasmin, Skinner, Michael A., Goodfellow, Ian G.
Formato: Texto
Lenguaje:English
Publicado: Society for General Microbiology 2007
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2884977/
https://www.ncbi.nlm.nih.gov/pubmed/17622609
http://dx.doi.org/10.1099/vir.0.82940-0
_version_ 1782182339360587776
author Chaudhry, Yasmin
Skinner, Michael A.
Goodfellow, Ian G.
author_facet Chaudhry, Yasmin
Skinner, Michael A.
Goodfellow, Ian G.
author_sort Chaudhry, Yasmin
collection PubMed
description Despite the significant disease burden caused by human norovirus infection, an efficient tissue-culture system for these viruses remains elusive. Murine norovirus (MNV) is an ideal surrogate for the study of norovirus biology, as the virus replicates efficiently in tissue culture and a low-cost animal model is readily available. In this report, a reverse-genetics system for MNV is described, using a fowlpox virus (FWPV) recombinant expressing T7 RNA polymerase to recover genetically defined MNV in tissue culture for the first time. These studies demonstrated that approaches that have proved successful for other members of the family Caliciviridae failed to lead to recovery of MNV. This was due to our observation that vaccinia virus infection had a negative effect on MNV replication. In contrast, FWPV infection had no deleterious effect and allowed the recovery of infectious MNV from cells previously transfected with MNV cDNA constructs. These studies also indicated that the nature of the 3′-terminal nucleotide is critical for efficient virus recovery and that inclusion of a hepatitis delta virus ribozyme at the 3′ end can increase the efficiency with which virus is recovered. This system now allows the recovery of genetically defined noroviruses and will facilitate the analysis of the effects of genetic variation on norovirus pathogenesis.
format Text
id pubmed-2884977
institution National Center for Biotechnology Information
language English
publishDate 2007
publisher Society for General Microbiology
record_format MEDLINE/PubMed
spelling pubmed-28849772010-07-06 Recovery of genetically defined murine norovirus in tissue culture by using a fowlpox virus expressing T7 RNA polymerase Chaudhry, Yasmin Skinner, Michael A. Goodfellow, Ian G. J Gen Virol Jgv Direct Despite the significant disease burden caused by human norovirus infection, an efficient tissue-culture system for these viruses remains elusive. Murine norovirus (MNV) is an ideal surrogate for the study of norovirus biology, as the virus replicates efficiently in tissue culture and a low-cost animal model is readily available. In this report, a reverse-genetics system for MNV is described, using a fowlpox virus (FWPV) recombinant expressing T7 RNA polymerase to recover genetically defined MNV in tissue culture for the first time. These studies demonstrated that approaches that have proved successful for other members of the family Caliciviridae failed to lead to recovery of MNV. This was due to our observation that vaccinia virus infection had a negative effect on MNV replication. In contrast, FWPV infection had no deleterious effect and allowed the recovery of infectious MNV from cells previously transfected with MNV cDNA constructs. These studies also indicated that the nature of the 3′-terminal nucleotide is critical for efficient virus recovery and that inclusion of a hepatitis delta virus ribozyme at the 3′ end can increase the efficiency with which virus is recovered. This system now allows the recovery of genetically defined noroviruses and will facilitate the analysis of the effects of genetic variation on norovirus pathogenesis. Society for General Microbiology 2007-08 /pmc/articles/PMC2884977/ /pubmed/17622609 http://dx.doi.org/10.1099/vir.0.82940-0 Text en Copyright © 2007, SGM http://creativecommons.org/licenses/by/2.5/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Jgv Direct
Chaudhry, Yasmin
Skinner, Michael A.
Goodfellow, Ian G.
Recovery of genetically defined murine norovirus in tissue culture by using a fowlpox virus expressing T7 RNA polymerase
title Recovery of genetically defined murine norovirus in tissue culture by using a fowlpox virus expressing T7 RNA polymerase
title_full Recovery of genetically defined murine norovirus in tissue culture by using a fowlpox virus expressing T7 RNA polymerase
title_fullStr Recovery of genetically defined murine norovirus in tissue culture by using a fowlpox virus expressing T7 RNA polymerase
title_full_unstemmed Recovery of genetically defined murine norovirus in tissue culture by using a fowlpox virus expressing T7 RNA polymerase
title_short Recovery of genetically defined murine norovirus in tissue culture by using a fowlpox virus expressing T7 RNA polymerase
title_sort recovery of genetically defined murine norovirus in tissue culture by using a fowlpox virus expressing t7 rna polymerase
topic Jgv Direct
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2884977/
https://www.ncbi.nlm.nih.gov/pubmed/17622609
http://dx.doi.org/10.1099/vir.0.82940-0
work_keys_str_mv AT chaudhryyasmin recoveryofgeneticallydefinedmurinenorovirusintissueculturebyusingafowlpoxvirusexpressingt7rnapolymerase
AT skinnermichaela recoveryofgeneticallydefinedmurinenorovirusintissueculturebyusingafowlpoxvirusexpressingt7rnapolymerase
AT goodfellowiang recoveryofgeneticallydefinedmurinenorovirusintissueculturebyusingafowlpoxvirusexpressingt7rnapolymerase