Transient Bluetongue virus serotype 8 capsid protein expression in Nicotiana benthamiana.
Identifieur interne : 000264 ( Main/Exploration ); précédent : 000263; suivant : 000265Transient Bluetongue virus serotype 8 capsid protein expression in Nicotiana benthamiana.
Auteurs : Albertha R. Van Zyl [Afrique du Sud] ; Ann E. Meyers [Afrique du Sud] ; Edward P. Rybicki [Afrique du Sud]Source :
- Biotechnology reports (Amsterdam, Netherlands) [ 2215-017X ] ; 2016.
Abstract
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.
DOI: 10.1016/j.btre.2015.12.001
PubMed: 28352588
PubMed Central: PMC5360979
Affiliations:
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Van Zyl</name><affiliation wicri:level="1"><nlm:affiliation>Department of Molecular and Cell Biology, University of Cape Town, Private Bag X3, Rondebosch 7700, South Africa.</nlm:affiliation><country xml:lang="fr">Afrique du Sud</country><wicri:regionArea>Department of Molecular and Cell Biology, University of Cape Town, Private Bag X3, Rondebosch 7700</wicri:regionArea><wicri:noRegion>Rondebosch 7700</wicri:noRegion></affiliation></author><author><name sortKey="Meyers, Ann E" sort="Meyers, Ann E" uniqKey="Meyers A" first="Ann E" last="Meyers">Ann E. Meyers</name><affiliation wicri:level="1"><nlm:affiliation>Department of Molecular and Cell Biology, University of Cape Town, Private Bag X3, Rondebosch 7700, South Africa.</nlm:affiliation><country xml:lang="fr">Afrique du Sud</country><wicri:regionArea>Department of Molecular and Cell Biology, University of Cape Town, Private Bag X3, Rondebosch 7700</wicri:regionArea><wicri:noRegion>Rondebosch 7700</wicri:noRegion></affiliation></author><author><name sortKey="Rybicki, Edward P" sort="Rybicki, Edward P" uniqKey="Rybicki E" first="Edward P" last="Rybicki">Edward P. Rybicki</name><affiliation wicri:level="1"><nlm:affiliation>Department of Molecular and Cell Biology, University of Cape Town, Private Bag X3, Rondebosch 7700, South Africa.</nlm:affiliation><country xml:lang="fr">Afrique du Sud</country><wicri:regionArea>Department of Molecular and Cell Biology, University of Cape Town, Private Bag X3, Rondebosch 7700</wicri:regionArea><wicri:noRegion>Rondebosch 7700</wicri:noRegion></affiliation></author></analytic><series><title level="j">Biotechnology reports (Amsterdam, Netherlands)</title><idno type="ISSN">2215-017X</idno><imprint><date when="2016" type="published">2016</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">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 <i>Agrobacterium</i>-mediated transient expression, an inexpensive production system very well suited to developing country use. Leaves infiltrated with recombinant pEAQ-<i>HT</i> vectors separately encoding the four BTV-8 capsid proteins produced more proteins than recombinant pTRA vectors. Plant expression using the pEAQ-<i>HT</i> 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. <i>In situ</i> 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.</div></front></TEI><pubmed><MedlineCitation Status="PubMed-not-MEDLINE" Owner="NLM"><PMID Version="1">28352588</PMID><DateRevised><Year>2020</Year><Month>10</Month><Day>01</Day></DateRevised><Article PubModel="Electronic-eCollection"><Journal><ISSN IssnType="Print">2215-017X</ISSN><JournalIssue CitedMedium="Print"><Volume>9</Volume><PubDate><Year>2016</Year><Month>Mar</Month></PubDate></JournalIssue><Title>Biotechnology reports (Amsterdam, Netherlands)</Title><ISOAbbreviation>Biotechnol Rep (Amst)</ISOAbbreviation></Journal><ArticleTitle>Transient Bluetongue virus serotype 8 capsid protein expression in <i>Nicotiana benthamiana</i>.</ArticleTitle><Pagination><MedlinePgn>15-24</MedlinePgn></Pagination><ELocationID EIdType="doi" ValidYN="Y">10.1016/j.btre.2015.12.001</ELocationID><Abstract><AbstractText>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 <i>Agrobacterium</i>-mediated transient expression, an inexpensive production system very well suited to developing country use. Leaves infiltrated with recombinant pEAQ-<i>HT</i> vectors separately encoding the four BTV-8 capsid proteins produced more proteins than recombinant pTRA vectors. Plant expression using the pEAQ-<i>HT</i> 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. <i>In situ</i> 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.</AbstractText></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>van Zyl</LastName><ForeName>Albertha R</ForeName><Initials>AR</Initials><AffiliationInfo><Affiliation>Department of Molecular and Cell Biology, University of Cape Town, Private Bag X3, Rondebosch 7700, South Africa.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Meyers</LastName><ForeName>Ann 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MajorTopicYN="N">Agroinfiltration</Keyword><Keyword MajorTopicYN="N">Bluetongue virus</Keyword><Keyword MajorTopicYN="N">Nicotiana benthamiana</Keyword><Keyword MajorTopicYN="N">Paracrystalline arrays</Keyword><Keyword MajorTopicYN="N">Virus-like particle</Keyword></KeywordList></MedlineCitation><PubmedData><History><PubMedPubDate PubStatus="received"><Year>2015</Year><Month>08</Month><Day>11</Day></PubMedPubDate><PubMedPubDate PubStatus="revised"><Year>2015</Year><Month>11</Month><Day>27</Day></PubMedPubDate><PubMedPubDate PubStatus="accepted"><Year>2015</Year><Month>12</Month><Day>01</Day></PubMedPubDate><PubMedPubDate PubStatus="entrez"><Year>2017</Year><Month>3</Month><Day>30</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="pubmed"><Year>2015</Year><Month>12</Month><Day>7</Day><Hour>0</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate 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IdType="pubmed">20422594</ArticleId></ArticleIdList></Reference></ReferenceList></PubmedData></pubmed><affiliations><list><country><li>Afrique du Sud</li></country></list><tree><country name="Afrique du Sud"><noRegion><name sortKey="Van Zyl, Albertha R" sort="Van Zyl, Albertha R" uniqKey="Van Zyl A" first="Albertha R" last="Van Zyl">Albertha R. Van Zyl</name></noRegion><name sortKey="Meyers, Ann E" sort="Meyers, Ann E" uniqKey="Meyers A" first="Ann E" last="Meyers">Ann E. Meyers</name><name sortKey="Rybicki, Edward P" sort="Rybicki, Edward P" uniqKey="Rybicki E" first="Edward P" last="Rybicki">Edward P. Rybicki</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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