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Transient Bluetongue virus serotype 8 capsid protein expression in Nicotiana benthamiana
Bluetongue virus (BTV) causes severe disease in domestic and wild ruminants, and has recently caused several outbreaks in Europe. Current vaccines include live-attenuated and inactivated viruses; while these are effective, there is risk of reversion to virulence by mutation or reassortment with wild...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier
2015
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5360979/ https://www.ncbi.nlm.nih.gov/pubmed/28352588 http://dx.doi.org/10.1016/j.btre.2015.12.001 |
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author | van Zyl, Albertha R. Meyers, Ann E. Rybicki, Edward P. |
author_facet | van Zyl, Albertha R. Meyers, Ann E. Rybicki, Edward P. |
author_sort | van Zyl, Albertha R. |
collection | PubMed |
description | Bluetongue virus (BTV) causes severe disease in domestic and wild ruminants, and has recently caused several outbreaks in Europe. Current vaccines include live-attenuated and inactivated viruses; while these are effective, there is risk of reversion to virulence by mutation or reassortment with wild type viruses. Subunit or virus-like particle (VLP) vaccines are safer options: VLP vaccines produced in insect cells by expression of the four BTV capsid proteins are protective against challenge; however, this is a costly production method. We investigated production of BTV VLPs in plants via Agrobacterium-mediated transient expression, an inexpensive production system very well suited to developing country use. Leaves infiltrated with recombinant pEAQ-HT vectors separately encoding the four BTV-8 capsid proteins produced more proteins than recombinant pTRA vectors. Plant expression using the pEAQ-HT vector resulted in both BTV-8 core-like particles (CLPs) and VLPs; differentially controlling the concentration of infiltrated bacteria significantly influenced yield of the VLPs. In situ localisation of assembled particles was investigated by using transmission electron microscopy (TEM) and it was shown that a mixed population of core-like particles (CLPs, consisting of VP3 and VP7) and VLPs were present as paracrystalline arrays in the cytoplasm of plant cells co-expressing all four capsid proteins. |
format | Online Article Text |
id | pubmed-5360979 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | Elsevier |
record_format | MEDLINE/PubMed |
spelling | pubmed-53609792017-03-28 Transient Bluetongue virus serotype 8 capsid protein expression in Nicotiana benthamiana van Zyl, Albertha R. Meyers, Ann E. Rybicki, Edward P. Biotechnol Rep (Amst) Article Bluetongue virus (BTV) causes severe disease in domestic and wild ruminants, and has recently caused several outbreaks in Europe. Current vaccines include live-attenuated and inactivated viruses; while these are effective, there is risk of reversion to virulence by mutation or reassortment with wild type viruses. Subunit or virus-like particle (VLP) vaccines are safer options: VLP vaccines produced in insect cells by expression of the four BTV capsid proteins are protective against challenge; however, this is a costly production method. We investigated production of BTV VLPs in plants via Agrobacterium-mediated transient expression, an inexpensive production system very well suited to developing country use. Leaves infiltrated with recombinant pEAQ-HT vectors separately encoding the four BTV-8 capsid proteins produced more proteins than recombinant pTRA vectors. Plant expression using the pEAQ-HT vector resulted in both BTV-8 core-like particles (CLPs) and VLPs; differentially controlling the concentration of infiltrated bacteria significantly influenced yield of the VLPs. In situ localisation of assembled particles was investigated by using transmission electron microscopy (TEM) and it was shown that a mixed population of core-like particles (CLPs, consisting of VP3 and VP7) and VLPs were present as paracrystalline arrays in the cytoplasm of plant cells co-expressing all four capsid proteins. Elsevier 2015-12-07 /pmc/articles/PMC5360979/ /pubmed/28352588 http://dx.doi.org/10.1016/j.btre.2015.12.001 Text en © 2015 The Authors http://creativecommons.org/licenses/by-nc-nd/4.0/ This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). |
spellingShingle | Article van Zyl, Albertha R. Meyers, Ann E. Rybicki, Edward P. Transient Bluetongue virus serotype 8 capsid protein expression in Nicotiana benthamiana |
title | Transient Bluetongue virus serotype 8 capsid protein expression in Nicotiana benthamiana |
title_full | Transient Bluetongue virus serotype 8 capsid protein expression in Nicotiana benthamiana |
title_fullStr | Transient Bluetongue virus serotype 8 capsid protein expression in Nicotiana benthamiana |
title_full_unstemmed | Transient Bluetongue virus serotype 8 capsid protein expression in Nicotiana benthamiana |
title_short | Transient Bluetongue virus serotype 8 capsid protein expression in Nicotiana benthamiana |
title_sort | transient bluetongue virus serotype 8 capsid protein expression in nicotiana benthamiana |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5360979/ https://www.ncbi.nlm.nih.gov/pubmed/28352588 http://dx.doi.org/10.1016/j.btre.2015.12.001 |
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