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Assembly of Recombinant Israeli Acute Paralysis Virus Capsids

The dicistrovirus Israeli Acute Paralysis Virus (IAPV) has been implicated in the worldwide decline of honey bees. Studies of IAPV and many other bee viruses in pure culture are restricted by available isolates and permissive cell culture. Here we show that coupling the IAPV major structural precurs...

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Detalles Bibliográficos
Autores principales: Ren, Junyuan, Cone, Abigail, Willmot, Rebecca, Jones, Ian M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4143317/
https://www.ncbi.nlm.nih.gov/pubmed/25153716
http://dx.doi.org/10.1371/journal.pone.0105943
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author Ren, Junyuan
Cone, Abigail
Willmot, Rebecca
Jones, Ian M.
author_facet Ren, Junyuan
Cone, Abigail
Willmot, Rebecca
Jones, Ian M.
author_sort Ren, Junyuan
collection PubMed
description The dicistrovirus Israeli Acute Paralysis Virus (IAPV) has been implicated in the worldwide decline of honey bees. Studies of IAPV and many other bee viruses in pure culture are restricted by available isolates and permissive cell culture. Here we show that coupling the IAPV major structural precursor protein ORF2 to its cognate 3C-like processing enzyme results in processing of the precursor to the individual structural proteins in a number of insect cell lines following expression by a recombinant baculovirus. The efficiency of expression is influenced by the level of IAPV 3C protein and moderation of its activity is required for optimal expression. The mature IAPV structural proteins assembled into empty capsids that migrated as particles on sucrose velocity gradients and showed typical dicistrovirus like morphology when examined by electron microscopy. Monoclonal antibodies raised to recombinant capsids were configured into a diagnostic test specific for the presence of IAPV. Recombinant capsids for each of the many bee viruses within the picornavirus family may provide virus specific reagents for the on-going investigation of the causes of honeybee loss.
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spelling pubmed-41433172014-08-27 Assembly of Recombinant Israeli Acute Paralysis Virus Capsids Ren, Junyuan Cone, Abigail Willmot, Rebecca Jones, Ian M. PLoS One Research Article The dicistrovirus Israeli Acute Paralysis Virus (IAPV) has been implicated in the worldwide decline of honey bees. Studies of IAPV and many other bee viruses in pure culture are restricted by available isolates and permissive cell culture. Here we show that coupling the IAPV major structural precursor protein ORF2 to its cognate 3C-like processing enzyme results in processing of the precursor to the individual structural proteins in a number of insect cell lines following expression by a recombinant baculovirus. The efficiency of expression is influenced by the level of IAPV 3C protein and moderation of its activity is required for optimal expression. The mature IAPV structural proteins assembled into empty capsids that migrated as particles on sucrose velocity gradients and showed typical dicistrovirus like morphology when examined by electron microscopy. Monoclonal antibodies raised to recombinant capsids were configured into a diagnostic test specific for the presence of IAPV. Recombinant capsids for each of the many bee viruses within the picornavirus family may provide virus specific reagents for the on-going investigation of the causes of honeybee loss. Public Library of Science 2014-08-25 /pmc/articles/PMC4143317/ /pubmed/25153716 http://dx.doi.org/10.1371/journal.pone.0105943 Text en © 2014 Ren et al http://creativecommons.org/licenses/by/4.0/ 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 author and source are properly credited.
spellingShingle Research Article
Ren, Junyuan
Cone, Abigail
Willmot, Rebecca
Jones, Ian M.
Assembly of Recombinant Israeli Acute Paralysis Virus Capsids
title Assembly of Recombinant Israeli Acute Paralysis Virus Capsids
title_full Assembly of Recombinant Israeli Acute Paralysis Virus Capsids
title_fullStr Assembly of Recombinant Israeli Acute Paralysis Virus Capsids
title_full_unstemmed Assembly of Recombinant Israeli Acute Paralysis Virus Capsids
title_short Assembly of Recombinant Israeli Acute Paralysis Virus Capsids
title_sort assembly of recombinant israeli acute paralysis virus capsids
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4143317/
https://www.ncbi.nlm.nih.gov/pubmed/25153716
http://dx.doi.org/10.1371/journal.pone.0105943
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