AgrobacTransV1, Main, Exploration, bibRecord, 000181

Expression of Rift Valley fever virus N-protein in Nicotiana benthamiana for use as a diagnostic antigen.

Identifieur interne : 000181 ( Main/Exploration ); précédent : 000180; suivant : 000182

Expression of Rift Valley fever virus N-protein in Nicotiana benthamiana for use as a diagnostic antigen.

Auteurs : Sandiswa Mbewana [Afrique du Sud] ; Ann E. Meyers [Afrique du Sud] ; Brandon Weber [Afrique du Sud] ; Vuyokazi Mareledwane [Afrique du Sud] ; Maryke L. Ferreira [Afrique du Sud] ; Phelix A O. Majiwa [Afrique du Sud] ; Edward P. Rybicki [Afrique du Sud]

Source :

BMC biotechnology [ 1472-6750 ] ; 2018.

RBID : pubmed:30537953

Descripteurs français

KwdFr :
- Animaux (MeSH), Antigènes viraux (génétique), Antigènes viraux (métabolisme), Antigènes viraux (sang), Expression des gènes (MeSH), Fièvre de la Vallée du Rift (diagnostic), Fièvre de la Vallée du Rift (sang), Fièvre de la Vallée du Rift (virologie), Maladies des ovins (diagnostic), Maladies des ovins (sang), Maladies des ovins (virologie), Ovis (MeSH), Protéines nucléocapside (génétique), Protéines nucléocapside (métabolisme), Protéines nucléocapside (sang), Tabac (génétique), Tabac (métabolisme), Test ELISA (méthodes), Virus de la fièvre de la vallée du Rift (génétique), Virus de la fièvre de la vallée du Rift (métabolisme).
MESH :
- diagnostic : Fièvre de la Vallée du Rift, Maladies des ovins.
- génétique : Antigènes viraux, Protéines nucléocapside, Tabac, Virus de la fièvre de la vallée du Rift.
- métabolisme : Antigènes viraux, Protéines nucléocapside, Tabac, Virus de la fièvre de la vallée du Rift.
- méthodes : Test ELISA.
- sang : Antigènes viraux, Fièvre de la Vallée du Rift, Maladies des ovins, Protéines nucléocapside.
- virologie : Fièvre de la Vallée du Rift, Maladies des ovins.
- Animaux, Expression des gènes, Ovis.

English descriptors

KwdEn :
- Animals (MeSH), Antigens, Viral (blood), Antigens, Viral (genetics), Antigens, Viral (metabolism), Enzyme-Linked Immunosorbent Assay (methods), Gene Expression (MeSH), Nucleocapsid Proteins (blood), Nucleocapsid Proteins (genetics), Nucleocapsid Proteins (metabolism), Rift Valley Fever (blood), Rift Valley Fever (diagnosis), Rift Valley Fever (virology), Rift Valley fever virus (genetics), Rift Valley fever virus (metabolism), Sheep (MeSH), Sheep Diseases (blood), Sheep Diseases (diagnosis), Sheep Diseases (virology), Tobacco (genetics), Tobacco (metabolism).
MESH :
- chemical , blood : Antigens, Viral, Nucleocapsid Proteins.
- chemical , genetics : Antigens, Viral, Nucleocapsid Proteins.
- chemical , metabolism : Antigens, Viral, Nucleocapsid Proteins.
- blood : Rift Valley Fever, Sheep Diseases.
- diagnosis : Rift Valley Fever, Sheep Diseases.
- genetics : Rift Valley fever virus, Tobacco.
- metabolism : Rift Valley fever virus, Tobacco.
- methods : Enzyme-Linked Immunosorbent Assay.
- virology : Rift Valley Fever, Sheep Diseases.
- Animals, Gene Expression, Sheep.

Abstract

BACKGROUND

Rift Valley fever virus (RVFV), the causative agent of Rift Valley fever, is an enveloped single-stranded negative-sense RNA virus in the genus Phlebovirus, family Bunyaviridae. The virus is spread by infected mosquitoes and affects ruminants and humans, causing abortion storms in pregnant ruminants, high neonatal mortality in animals, and morbidity and occasional fatalities in humans. The disease is endemic in parts of Africa and the Arabian Peninsula, but is described as emerging due to the wide range of mosquitoes that could spread the disease into non-endemic regions. There are different tests for determining whether animals are infected with or have been exposed to RVFV. The most common serological test is antibody ELISA, which detects host immunoglobulins M or G produced specifically in response to infection with RVFV. The presence of antibodies to RVFV nucleocapsid protein (N-protein) is among the best indicators of RVFV exposure in animals. This work describes an investigation of the feasibility of producing a recombinant N-protein in Nicotiana benthamiana and using it in an ELISA.

RESULTS

The human-codon optimised RVFV N-protein was successfully expressed in N. benthamiana via Agrobacterium-mediated infiltration of leaves. The recombinant protein was detected as monomers and dimers with maximum protein yields calculated to be 500-558 mg/kg of fresh plant leaves. The identity of the protein was confirmed by liquid chromatography-mass spectrometry (LC-MS) resulting in 87.35% coverage, with 264 unique peptides. Transmission electron microscopy revealed that the protein forms ring structures of ~ 10 nm in diameter. Preliminary data revealed that the protein could successfully differentiate between sera of RVFV-infected sheep and from sera of those not infected with the virus.

CONCLUSIONS

To the best of our knowledge this is the first study demonstrating the successful production of RVFV N-protein as a diagnostic reagent by Agrobacterium-mediated transient heterologous expression in N. benthamiana. Preliminary testing of the antigen showed its ability to distinguish RVFV-positive animal sera from RVFV negative animal sera when used in an enzyme linked immunosorbent assay (ELISA). The cost-effective, scalable and simple production method has great potential for use in developing countries where rapid diagnosis of RVFV is necessary.

DOI: 10.1186/s12896-018-0489-z
PubMed: 30537953
PubMed Central: PMC6290525

Affiliations:

Afrique du Sud

Links toward previous steps (curation, corpus...)

to stream Main, to step Corpus: 000135
to stream Main, to step Curation: 000135

Le document en format XML

<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Expression of Rift Valley fever virus N-protein in Nicotiana benthamiana for use as a diagnostic antigen.</title>
<author><name sortKey="Mbewana, Sandiswa" sort="Mbewana, Sandiswa" uniqKey="Mbewana S" first="Sandiswa" last="Mbewana">Sandiswa Mbewana</name>
<affiliation wicri:level="1"><nlm:affiliation>Biopharming Research Unit, Department of Molecular and Cell Biology, University of Cape Town, 6503200115084, Rondebosch, Cape Town, 7700, South Africa.</nlm:affiliation>
<country xml:lang="fr">Afrique du Sud</country>
<wicri:regionArea>Biopharming Research Unit, Department of Molecular and Cell Biology, University of Cape Town, 6503200115084, Rondebosch, Cape Town, 7700</wicri:regionArea>
<wicri:noRegion>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>Biopharming Research Unit, Department of Molecular and Cell Biology, University of Cape Town, 6503200115084, Rondebosch, Cape Town, 7700, South Africa. ann.meyers@uct.ac.za.</nlm:affiliation>
<country xml:lang="fr">Afrique du Sud</country>
<wicri:regionArea>Biopharming Research Unit, Department of Molecular and Cell Biology, University of Cape Town, 6503200115084, Rondebosch, Cape Town, 7700</wicri:regionArea>
<wicri:noRegion>7700</wicri:noRegion>
</affiliation>
</author>
<author><name sortKey="Weber, Brandon" sort="Weber, Brandon" uniqKey="Weber B" first="Brandon" last="Weber">Brandon Weber</name>
<affiliation wicri:level="1"><nlm:affiliation>Structural Biology Research Unit, University of Cape Town, P Bag X3, Rondebosch, 7700, South Africa.</nlm:affiliation>
<country xml:lang="fr">Afrique du Sud</country>
<wicri:regionArea>Structural Biology Research Unit, University of Cape Town, P Bag X3, Rondebosch, 7700</wicri:regionArea>
<wicri:noRegion>7700</wicri:noRegion>
</affiliation>
</author>
<author><name sortKey="Mareledwane, Vuyokazi" sort="Mareledwane, Vuyokazi" uniqKey="Mareledwane V" first="Vuyokazi" last="Mareledwane">Vuyokazi Mareledwane</name>
<affiliation wicri:level="1"><nlm:affiliation>ARC-Onderstepoort Veterinary Institute, 100 Old Southpan Road, Onderstepoort, 0110, South Africa.</nlm:affiliation>
<country xml:lang="fr">Afrique du Sud</country>
<wicri:regionArea>ARC-Onderstepoort Veterinary Institute, 100 Old Southpan Road, Onderstepoort, 0110</wicri:regionArea>
<wicri:noRegion>0110</wicri:noRegion>
</affiliation>
</author>
<author><name sortKey="Ferreira, Maryke L" sort="Ferreira, Maryke L" uniqKey="Ferreira M" first="Maryke L" last="Ferreira">Maryke L. Ferreira</name>
<affiliation wicri:level="1"><nlm:affiliation>ARC-Onderstepoort Veterinary Institute, 100 Old Southpan Road, Onderstepoort, 0110, South Africa.</nlm:affiliation>
<country xml:lang="fr">Afrique du Sud</country>
<wicri:regionArea>ARC-Onderstepoort Veterinary Institute, 100 Old Southpan Road, Onderstepoort, 0110</wicri:regionArea>
<wicri:noRegion>0110</wicri:noRegion>
</affiliation>
</author>
<author><name sortKey="Majiwa, Phelix A O" sort="Majiwa, Phelix A O" uniqKey="Majiwa P" first="Phelix A O" last="Majiwa">Phelix A O. Majiwa</name>
<affiliation wicri:level="1"><nlm:affiliation>ARC-Onderstepoort Veterinary Institute, 100 Old Southpan Road, Onderstepoort, 0110, South Africa.</nlm:affiliation>
<country xml:lang="fr">Afrique du Sud</country>
<wicri:regionArea>ARC-Onderstepoort Veterinary Institute, 100 Old Southpan Road, Onderstepoort, 0110</wicri:regionArea>
<wicri:noRegion>0110</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>Biopharming Research Unit, Department of Molecular and Cell Biology, University of Cape Town, 6503200115084, Rondebosch, Cape Town, 7700, South Africa.</nlm:affiliation>
<country xml:lang="fr">Afrique du Sud</country>
<wicri:regionArea>Biopharming Research Unit, Department of Molecular and Cell Biology, University of Cape Town, 6503200115084, Rondebosch, Cape Town, 7700</wicri:regionArea>
<wicri:noRegion>7700</wicri:noRegion>
</affiliation>
<affiliation wicri:level="1"><nlm:affiliation>Institute of Infectious Disease and Molecular Medicine, University of Cape Town, Anzio Road, Observatory, Cape Town, 7925, South Africa.</nlm:affiliation>
<country xml:lang="fr">Afrique du Sud</country>
<wicri:regionArea>Institute of Infectious Disease and Molecular Medicine, University of Cape Town, Anzio Road, Observatory, Cape Town, 7925</wicri:regionArea>
<wicri:noRegion>7925</wicri:noRegion>
</affiliation>
</author>
</titleStmt>
<publicationStmt><idno type="wicri:source">PubMed</idno>
<date when="2018">2018</date>
<idno type="RBID">pubmed:30537953</idno>
<idno type="pmid">30537953</idno>
<idno type="doi">10.1186/s12896-018-0489-z</idno>
<idno type="pmc">PMC6290525</idno>
<idno type="wicri:Area/Main/Corpus">000135</idno>
<idno type="wicri:explorRef" wicri:stream="Main" wicri:step="Corpus" wicri:corpus="PubMed">000135</idno>
<idno type="wicri:Area/Main/Curation">000135</idno>
<idno type="wicri:explorRef" wicri:stream="Main" wicri:step="Curation">000135</idno>
<idno type="wicri:Area/Main/Exploration">000135</idno>
</publicationStmt>
<sourceDesc><biblStruct><analytic><title xml:lang="en">Expression of Rift Valley fever virus N-protein in Nicotiana benthamiana for use as a diagnostic antigen.</title>
<author><name sortKey="Mbewana, Sandiswa" sort="Mbewana, Sandiswa" uniqKey="Mbewana S" first="Sandiswa" last="Mbewana">Sandiswa Mbewana</name>
<affiliation wicri:level="1"><nlm:affiliation>Biopharming Research Unit, Department of Molecular and Cell Biology, University of Cape Town, 6503200115084, Rondebosch, Cape Town, 7700, South Africa.</nlm:affiliation>
<country xml:lang="fr">Afrique du Sud</country>
<wicri:regionArea>Biopharming Research Unit, Department of Molecular and Cell Biology, University of Cape Town, 6503200115084, Rondebosch, Cape Town, 7700</wicri:regionArea>
<wicri:noRegion>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>Biopharming Research Unit, Department of Molecular and Cell Biology, University of Cape Town, 6503200115084, Rondebosch, Cape Town, 7700, South Africa. ann.meyers@uct.ac.za.</nlm:affiliation>
<country xml:lang="fr">Afrique du Sud</country>
<wicri:regionArea>Biopharming Research Unit, Department of Molecular and Cell Biology, University of Cape Town, 6503200115084, Rondebosch, Cape Town, 7700</wicri:regionArea>
<wicri:noRegion>7700</wicri:noRegion>
</affiliation>
</author>
<author><name sortKey="Weber, Brandon" sort="Weber, Brandon" uniqKey="Weber B" first="Brandon" last="Weber">Brandon Weber</name>
<affiliation wicri:level="1"><nlm:affiliation>Structural Biology Research Unit, University of Cape Town, P Bag X3, Rondebosch, 7700, South Africa.</nlm:affiliation>
<country xml:lang="fr">Afrique du Sud</country>
<wicri:regionArea>Structural Biology Research Unit, University of Cape Town, P Bag X3, Rondebosch, 7700</wicri:regionArea>
<wicri:noRegion>7700</wicri:noRegion>
</affiliation>
</author>
<author><name sortKey="Mareledwane, Vuyokazi" sort="Mareledwane, Vuyokazi" uniqKey="Mareledwane V" first="Vuyokazi" last="Mareledwane">Vuyokazi Mareledwane</name>
<affiliation wicri:level="1"><nlm:affiliation>ARC-Onderstepoort Veterinary Institute, 100 Old Southpan Road, Onderstepoort, 0110, South Africa.</nlm:affiliation>
<country xml:lang="fr">Afrique du Sud</country>
<wicri:regionArea>ARC-Onderstepoort Veterinary Institute, 100 Old Southpan Road, Onderstepoort, 0110</wicri:regionArea>
<wicri:noRegion>0110</wicri:noRegion>
</affiliation>
</author>
<author><name sortKey="Ferreira, Maryke L" sort="Ferreira, Maryke L" uniqKey="Ferreira M" first="Maryke L" last="Ferreira">Maryke L. Ferreira</name>
<affiliation wicri:level="1"><nlm:affiliation>ARC-Onderstepoort Veterinary Institute, 100 Old Southpan Road, Onderstepoort, 0110, South Africa.</nlm:affiliation>
<country xml:lang="fr">Afrique du Sud</country>
<wicri:regionArea>ARC-Onderstepoort Veterinary Institute, 100 Old Southpan Road, Onderstepoort, 0110</wicri:regionArea>
<wicri:noRegion>0110</wicri:noRegion>
</affiliation>
</author>
<author><name sortKey="Majiwa, Phelix A O" sort="Majiwa, Phelix A O" uniqKey="Majiwa P" first="Phelix A O" last="Majiwa">Phelix A O. Majiwa</name>
<affiliation wicri:level="1"><nlm:affiliation>ARC-Onderstepoort Veterinary Institute, 100 Old Southpan Road, Onderstepoort, 0110, South Africa.</nlm:affiliation>
<country xml:lang="fr">Afrique du Sud</country>
<wicri:regionArea>ARC-Onderstepoort Veterinary Institute, 100 Old Southpan Road, Onderstepoort, 0110</wicri:regionArea>
<wicri:noRegion>0110</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>Biopharming Research Unit, Department of Molecular and Cell Biology, University of Cape Town, 6503200115084, Rondebosch, Cape Town, 7700, South Africa.</nlm:affiliation>
<country xml:lang="fr">Afrique du Sud</country>
<wicri:regionArea>Biopharming Research Unit, Department of Molecular and Cell Biology, University of Cape Town, 6503200115084, Rondebosch, Cape Town, 7700</wicri:regionArea>
<wicri:noRegion>7700</wicri:noRegion>
</affiliation>
<affiliation wicri:level="1"><nlm:affiliation>Institute of Infectious Disease and Molecular Medicine, University of Cape Town, Anzio Road, Observatory, Cape Town, 7925, South Africa.</nlm:affiliation>
<country xml:lang="fr">Afrique du Sud</country>
<wicri:regionArea>Institute of Infectious Disease and Molecular Medicine, University of Cape Town, Anzio Road, Observatory, Cape Town, 7925</wicri:regionArea>
<wicri:noRegion>7925</wicri:noRegion>
</affiliation>
</author>
</analytic>
<series><title level="j">BMC biotechnology</title>
<idno type="eISSN">1472-6750</idno>
<imprint><date when="2018" type="published">2018</date>
</imprint>
</series>
</biblStruct>
</sourceDesc>
</fileDesc>
<profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Animals (MeSH)</term>
<term>Antigens, Viral (blood)</term>
<term>Antigens, Viral (genetics)</term>
<term>Antigens, Viral (metabolism)</term>
<term>Enzyme-Linked Immunosorbent Assay (methods)</term>
<term>Gene Expression (MeSH)</term>
<term>Nucleocapsid Proteins (blood)</term>
<term>Nucleocapsid Proteins (genetics)</term>
<term>Nucleocapsid Proteins (metabolism)</term>
<term>Rift Valley Fever (blood)</term>
<term>Rift Valley Fever (diagnosis)</term>
<term>Rift Valley Fever (virology)</term>
<term>Rift Valley fever virus (genetics)</term>
<term>Rift Valley fever virus (metabolism)</term>
<term>Sheep (MeSH)</term>
<term>Sheep Diseases (blood)</term>
<term>Sheep Diseases (diagnosis)</term>
<term>Sheep Diseases (virology)</term>
<term>Tobacco (genetics)</term>
<term>Tobacco (metabolism)</term>
</keywords>
<keywords scheme="KwdFr" xml:lang="fr"><term>Animaux (MeSH)</term>
<term>Antigènes viraux (génétique)</term>
<term>Antigènes viraux (métabolisme)</term>
<term>Antigènes viraux (sang)</term>
<term>Expression des gènes (MeSH)</term>
<term>Fièvre de la Vallée du Rift (diagnostic)</term>
<term>Fièvre de la Vallée du Rift (sang)</term>
<term>Fièvre de la Vallée du Rift (virologie)</term>
<term>Maladies des ovins (diagnostic)</term>
<term>Maladies des ovins (sang)</term>
<term>Maladies des ovins (virologie)</term>
<term>Ovis (MeSH)</term>
<term>Protéines nucléocapside (génétique)</term>
<term>Protéines nucléocapside (métabolisme)</term>
<term>Protéines nucléocapside (sang)</term>
<term>Tabac (génétique)</term>
<term>Tabac (métabolisme)</term>
<term>Test ELISA (méthodes)</term>
<term>Virus de la fièvre de la vallée du Rift (génétique)</term>
<term>Virus de la fièvre de la vallée du Rift (métabolisme)</term>
</keywords>
<keywords scheme="MESH" type="chemical" qualifier="blood" xml:lang="en"><term>Antigens, Viral</term>
<term>Nucleocapsid Proteins</term>
</keywords>
<keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>Antigens, Viral</term>
<term>Nucleocapsid Proteins</term>
</keywords>
<keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>Antigens, Viral</term>
<term>Nucleocapsid Proteins</term>
</keywords>
<keywords scheme="MESH" qualifier="blood" xml:lang="en"><term>Rift Valley Fever</term>
<term>Sheep Diseases</term>
</keywords>
<keywords scheme="MESH" qualifier="diagnosis" xml:lang="en"><term>Rift Valley Fever</term>
<term>Sheep Diseases</term>
</keywords>
<keywords scheme="MESH" qualifier="diagnostic" xml:lang="fr"><term>Fièvre de la Vallée du Rift</term>
<term>Maladies des ovins</term>
</keywords>
<keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Rift Valley fever virus</term>
<term>Tobacco</term>
</keywords>
<keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>Antigènes viraux</term>
<term>Protéines nucléocapside</term>
<term>Tabac</term>
<term>Virus de la fièvre de la vallée du Rift</term>
</keywords>
<keywords scheme="MESH" qualifier="metabolism" xml:lang="en"><term>Rift Valley fever virus</term>
<term>Tobacco</term>
</keywords>
<keywords scheme="MESH" qualifier="methods" xml:lang="en"><term>Enzyme-Linked Immunosorbent Assay</term>
</keywords>
<keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>Antigènes viraux</term>
<term>Protéines nucléocapside</term>
<term>Tabac</term>
<term>Virus de la fièvre de la vallée du Rift</term>
</keywords>
<keywords scheme="MESH" qualifier="méthodes" xml:lang="fr"><term>Test ELISA</term>
</keywords>
<keywords scheme="MESH" qualifier="sang" xml:lang="fr"><term>Antigènes viraux</term>
<term>Fièvre de la Vallée du Rift</term>
<term>Maladies des ovins</term>
<term>Protéines nucléocapside</term>
</keywords>
<keywords scheme="MESH" qualifier="virologie" xml:lang="fr"><term>Fièvre de la Vallée du Rift</term>
<term>Maladies des ovins</term>
</keywords>
<keywords scheme="MESH" qualifier="virology" xml:lang="en"><term>Rift Valley Fever</term>
<term>Sheep Diseases</term>
</keywords>
<keywords scheme="MESH" xml:lang="en"><term>Animals</term>
<term>Gene Expression</term>
<term>Sheep</term>
</keywords>
<keywords scheme="MESH" xml:lang="fr"><term>Animaux</term>
<term>Expression des gènes</term>
<term>Ovis</term>
</keywords>
</textClass>
</profileDesc>
</teiHeader>
<front><div type="abstract" xml:lang="en"><p><b>BACKGROUND</b>
</p>
<p>Rift Valley fever virus (RVFV), the causative agent of Rift Valley fever, is an enveloped single-stranded negative-sense RNA virus in the genus Phlebovirus, family Bunyaviridae. The virus is spread by infected mosquitoes and affects ruminants and humans, causing abortion storms in pregnant ruminants, high neonatal mortality in animals, and morbidity and occasional fatalities in humans. The disease is endemic in parts of Africa and the Arabian Peninsula, but is described as emerging due to the wide range of mosquitoes that could spread the disease into non-endemic regions. There are different tests for determining whether animals are infected with or have been exposed to RVFV. The most common serological test is antibody ELISA, which detects host immunoglobulins M or G produced specifically in response to infection with RVFV. The presence of antibodies to RVFV nucleocapsid protein (N-protein) is among the best indicators of RVFV exposure in animals. This work describes an investigation of the feasibility of producing a recombinant N-protein in Nicotiana benthamiana and using it in an ELISA.</p>
</div>
<div type="abstract" xml:lang="en"><p><b>RESULTS</b>
</p>
<p>The human-codon optimised RVFV N-protein was successfully expressed in N. benthamiana via Agrobacterium-mediated infiltration of leaves. The recombinant protein was detected as monomers and dimers with maximum protein yields calculated to be 500-558 mg/kg of fresh plant leaves. The identity of the protein was confirmed by liquid chromatography-mass spectrometry (LC-MS) resulting in 87.35% coverage, with 264 unique peptides. Transmission electron microscopy revealed that the protein forms ring structures of ~ 10 nm in diameter. Preliminary data revealed that the protein could successfully differentiate between sera of RVFV-infected sheep and from sera of those not infected with the virus.</p>
</div>
<div type="abstract" xml:lang="en"><p><b>CONCLUSIONS</b>
</p>
<p>To the best of our knowledge this is the first study demonstrating the successful production of RVFV N-protein as a diagnostic reagent by Agrobacterium-mediated transient heterologous expression in N. benthamiana. Preliminary testing of the antigen showed its ability to distinguish RVFV-positive animal sera from RVFV negative animal sera when used in an enzyme linked immunosorbent assay (ELISA). The cost-effective, scalable and simple production method has great potential for use in developing countries where rapid diagnosis of RVFV is necessary.</p>
</div>
</front>
</TEI>
<pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">30537953</PMID>
<DateCompleted><Year>2019</Year>
<Month>08</Month>
<Day>08</Day>
</DateCompleted>
<DateRevised><Year>2020</Year>
<Month>03</Month>
<Day>09</Day>
</DateRevised>
<Article PubModel="Electronic"><Journal><ISSN IssnType="Electronic">1472-6750</ISSN>
<JournalIssue CitedMedium="Internet"><Volume>18</Volume>
<Issue>1</Issue>
<PubDate><Year>2018</Year>
<Month>12</Month>
<Day>11</Day>
</PubDate>
</JournalIssue>
<Title>BMC biotechnology</Title>
<ISOAbbreviation>BMC Biotechnol</ISOAbbreviation>
</Journal>
<ArticleTitle>Expression of Rift Valley fever virus N-protein in Nicotiana benthamiana for use as a diagnostic antigen.</ArticleTitle>
<Pagination><MedlinePgn>77</MedlinePgn>
</Pagination>
<ELocationID EIdType="doi" ValidYN="Y">10.1186/s12896-018-0489-z</ELocationID>
<Abstract><AbstractText Label="BACKGROUND">Rift Valley fever virus (RVFV), the causative agent of Rift Valley fever, is an enveloped single-stranded negative-sense RNA virus in the genus Phlebovirus, family Bunyaviridae. The virus is spread by infected mosquitoes and affects ruminants and humans, causing abortion storms in pregnant ruminants, high neonatal mortality in animals, and morbidity and occasional fatalities in humans. The disease is endemic in parts of Africa and the Arabian Peninsula, but is described as emerging due to the wide range of mosquitoes that could spread the disease into non-endemic regions. There are different tests for determining whether animals are infected with or have been exposed to RVFV. The most common serological test is antibody ELISA, which detects host immunoglobulins M or G produced specifically in response to infection with RVFV. The presence of antibodies to RVFV nucleocapsid protein (N-protein) is among the best indicators of RVFV exposure in animals. This work describes an investigation of the feasibility of producing a recombinant N-protein in Nicotiana benthamiana and using it in an ELISA.</AbstractText>
<AbstractText Label="RESULTS">The human-codon optimised RVFV N-protein was successfully expressed in N. benthamiana via Agrobacterium-mediated infiltration of leaves. The recombinant protein was detected as monomers and dimers with maximum protein yields calculated to be 500-558 mg/kg of fresh plant leaves. The identity of the protein was confirmed by liquid chromatography-mass spectrometry (LC-MS) resulting in 87.35% coverage, with 264 unique peptides. Transmission electron microscopy revealed that the protein forms ring structures of ~ 10 nm in diameter. Preliminary data revealed that the protein could successfully differentiate between sera of RVFV-infected sheep and from sera of those not infected with the virus.</AbstractText>
<AbstractText Label="CONCLUSIONS">To the best of our knowledge this is the first study demonstrating the successful production of RVFV N-protein as a diagnostic reagent by Agrobacterium-mediated transient heterologous expression in N. benthamiana. Preliminary testing of the antigen showed its ability to distinguish RVFV-positive animal sera from RVFV negative animal sera when used in an enzyme linked immunosorbent assay (ELISA). The cost-effective, scalable and simple production method has great potential for use in developing countries where rapid diagnosis of RVFV is necessary.</AbstractText>
</Abstract>
<AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Mbewana</LastName>
<ForeName>Sandiswa</ForeName>
<Initials>S</Initials>
<AffiliationInfo><Affiliation>Biopharming Research Unit, Department of Molecular and Cell Biology, University of Cape Town, 6503200115084, Rondebosch, Cape Town, 7700, South Africa.</Affiliation>
</AffiliationInfo>
</Author>
<Author ValidYN="Y"><LastName>Meyers</LastName>
<ForeName>Ann E</ForeName>
<Initials>AE</Initials>
<Identifier Source="ORCID">0000-0002-2116-7913</Identifier>
<AffiliationInfo><Affiliation>Biopharming Research Unit, Department of Molecular and Cell Biology, University of Cape Town, 6503200115084, Rondebosch, Cape Town, 7700, South Africa. ann.meyers@uct.ac.za.</Affiliation>
</AffiliationInfo>
</Author>
<Author ValidYN="Y"><LastName>Weber</LastName>
<ForeName>Brandon</ForeName>
<Initials>B</Initials>
<AffiliationInfo><Affiliation>Structural Biology Research Unit, University of Cape Town, P Bag X3, Rondebosch, 7700, South Africa.</Affiliation>
</AffiliationInfo>
</Author>
<Author ValidYN="Y"><LastName>Mareledwane</LastName>
<ForeName>Vuyokazi</ForeName>
<Initials>V</Initials>
<AffiliationInfo><Affiliation>ARC-Onderstepoort Veterinary Institute, 100 Old Southpan Road, Onderstepoort, 0110, South Africa.</Affiliation>
</AffiliationInfo>
</Author>
<Author ValidYN="Y"><LastName>Ferreira</LastName>
<ForeName>Maryke L</ForeName>
<Initials>ML</Initials>
<AffiliationInfo><Affiliation>ARC-Onderstepoort Veterinary Institute, 100 Old Southpan Road, Onderstepoort, 0110, South Africa.</Affiliation>
</AffiliationInfo>
</Author>
<Author ValidYN="Y"><LastName>Majiwa</LastName>
<ForeName>Phelix A O</ForeName>
<Initials>PAO</Initials>
<AffiliationInfo><Affiliation>ARC-Onderstepoort Veterinary Institute, 100 Old Southpan Road, Onderstepoort, 0110, South Africa.</Affiliation>
</AffiliationInfo>
</Author>
<Author ValidYN="Y"><LastName>Rybicki</LastName>
<ForeName>Edward P</ForeName>
<Initials>EP</Initials>
<AffiliationInfo><Affiliation>Biopharming Research Unit, Department of Molecular and Cell Biology, University of Cape Town, 6503200115084, Rondebosch, Cape Town, 7700, South Africa.</Affiliation>
</AffiliationInfo>
<AffiliationInfo><Affiliation>Institute of Infectious Disease and Molecular Medicine, University of Cape Town, Anzio Road, Observatory, Cape Town, 7925, South Africa.</Affiliation>
</AffiliationInfo>
</Author>
</AuthorList>
<Language>eng</Language>
<PublicationTypeList><PublicationType UI="D016428">Journal Article</PublicationType>
<PublicationType UI="D013485">Research Support, Non-U.S. Gov't</PublicationType>
</PublicationTypeList>
<ArticleDate DateType="Electronic"><Year>2018</Year>
<Month>12</Month>
<Day>11</Day>
</ArticleDate>
</Article>
<MedlineJournalInfo><Country>England</Country>
<MedlineTA>BMC Biotechnol</MedlineTA>
<NlmUniqueID>101088663</NlmUniqueID>
<ISSNLinking>1472-6750</ISSNLinking>
</MedlineJournalInfo>
<ChemicalList><Chemical><RegistryNumber>0</RegistryNumber>
<NameOfSubstance UI="D000956">Antigens, Viral</NameOfSubstance>
</Chemical>
<Chemical><RegistryNumber>0</RegistryNumber>
<NameOfSubstance UI="D019590">Nucleocapsid Proteins</NameOfSubstance>
</Chemical>
</ChemicalList>
<MeshHeadingList><MeshHeading><DescriptorName UI="D000818" MajorTopicYN="N">Animals</DescriptorName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D000956" MajorTopicYN="N">Antigens, Viral</DescriptorName>
<QualifierName UI="Q000097" MajorTopicYN="N">blood</QualifierName>
<QualifierName UI="Q000235" MajorTopicYN="Y">genetics</QualifierName>
<QualifierName UI="Q000378" MajorTopicYN="N">metabolism</QualifierName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D004797" MajorTopicYN="N">Enzyme-Linked Immunosorbent Assay</DescriptorName>
<QualifierName UI="Q000379" MajorTopicYN="N">methods</QualifierName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D015870" MajorTopicYN="N">Gene Expression</DescriptorName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D019590" MajorTopicYN="N">Nucleocapsid Proteins</DescriptorName>
<QualifierName UI="Q000097" MajorTopicYN="N">blood</QualifierName>
<QualifierName UI="Q000235" MajorTopicYN="Y">genetics</QualifierName>
<QualifierName UI="Q000378" MajorTopicYN="N">metabolism</QualifierName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D012295" MajorTopicYN="N">Rift Valley Fever</DescriptorName>
<QualifierName UI="Q000097" MajorTopicYN="N">blood</QualifierName>
<QualifierName UI="Q000175" MajorTopicYN="Y">diagnosis</QualifierName>
<QualifierName UI="Q000821" MajorTopicYN="N">virology</QualifierName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D012296" MajorTopicYN="N">Rift Valley fever virus</DescriptorName>
<QualifierName UI="Q000235" MajorTopicYN="Y">genetics</QualifierName>
<QualifierName UI="Q000378" MajorTopicYN="Y">metabolism</QualifierName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D012756" MajorTopicYN="N">Sheep</DescriptorName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D012757" MajorTopicYN="N">Sheep Diseases</DescriptorName>
<QualifierName UI="Q000097" MajorTopicYN="N">blood</QualifierName>
<QualifierName UI="Q000175" MajorTopicYN="Y">diagnosis</QualifierName>
<QualifierName UI="Q000821" MajorTopicYN="N">virology</QualifierName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D014026" MajorTopicYN="N">Tobacco</DescriptorName>
<QualifierName UI="Q000235" MajorTopicYN="Y">genetics</QualifierName>
<QualifierName UI="Q000378" MajorTopicYN="N">metabolism</QualifierName>
</MeshHeading>
</MeshHeadingList>
<KeywordList Owner="NOTNLM"><Keyword MajorTopicYN="Y">DIVA</Keyword>
<Keyword MajorTopicYN="Y">Diagnostic</Keyword>
<Keyword MajorTopicYN="Y">ELISA</Keyword>
<Keyword MajorTopicYN="Y">Molecular farming</Keyword>
<Keyword MajorTopicYN="Y">N-protein</Keyword>
<Keyword MajorTopicYN="Y">Plant production</Keyword>
<Keyword MajorTopicYN="Y">Rift Valley fever virus</Keyword>
</KeywordList>
</MedlineCitation>
<PubmedData><History><PubMedPubDate PubStatus="received"><Year>2017</Year>
<Month>11</Month>
<Day>29</Day>
</PubMedPubDate>
<PubMedPubDate PubStatus="accepted"><Year>2018</Year>
<Month>11</Month>
<Day>29</Day>
</PubMedPubDate>
<PubMedPubDate PubStatus="entrez"><Year>2018</Year>
<Month>12</Month>
<Day>13</Day>
<Hour>6</Hour>
<Minute>0</Minute>
</PubMedPubDate>
<PubMedPubDate PubStatus="pubmed"><Year>2018</Year>
<Month>12</Month>
<Day>13</Day>
<Hour>6</Hour>
<Minute>0</Minute>
</PubMedPubDate>
<PubMedPubDate PubStatus="medline"><Year>2019</Year>
<Month>8</Month>
<Day>9</Day>
<Hour>6</Hour>
<Minute>0</Minute>
</PubMedPubDate>
</History>
<PublicationStatus>epublish</PublicationStatus>
<ArticleIdList><ArticleId IdType="pubmed">30537953</ArticleId>
<ArticleId IdType="doi">10.1186/s12896-018-0489-z</ArticleId>
<ArticleId IdType="pii">10.1186/s12896-018-0489-z</ArticleId>
<ArticleId IdType="pmc">PMC6290525</ArticleId>
</ArticleIdList>
<ReferenceList><Reference><Citation>Protein Expr Purif. 2016 Nov;127:61-67</Citation>
<ArticleIdList><ArticleId IdType="pubmed">27402440</ArticleId>
</ArticleIdList>
</Reference>
<Reference><Citation>Transbound Emerg Dis. 2013 Nov;60 Suppl 2:24-30</Citation>
<ArticleIdList><ArticleId IdType="pubmed">24589098</ArticleId>
</ArticleIdList>
</Reference>
<Reference><Citation>BMC Biotechnol. 2012 Apr 26;12:14</Citation>
<ArticleIdList><ArticleId IdType="pubmed">22536810</ArticleId>
</ArticleIdList>
</Reference>
<Reference><Citation>Ecohealth. 2012 Jun;9(2):107-10</Citation>
<ArticleIdList><ArticleId IdType="pubmed">22777051</ArticleId>
</ArticleIdList>
</Reference>
<Reference><Citation>PLoS Pathog. 2011 May;7(5):e1002030</Citation>
<ArticleIdList><ArticleId IdType="pubmed">21589902</ArticleId>
</ArticleIdList>
</Reference>
<Reference><Citation>J Hyg (Lond). 1986 Oct;97(2):317-29</Citation>
<ArticleIdList><ArticleId IdType="pubmed">3537118</ArticleId>
</ArticleIdList>
</Reference>
<Reference><Citation>Biomed Res Int. 2014;2014:136419</Citation>
<ArticleIdList><ArticleId IdType="pubmed">24745008</ArticleId>
</ArticleIdList>
</Reference>
<Reference><Citation>J Virol. 2005 Sep;79(18):11974-80</Citation>
<ArticleIdList><ArticleId IdType="pubmed">16140773</ArticleId>
</ArticleIdList>
</Reference>
<Reference><Citation>J Clin Microbiol. 2001 Dec;39(12):4456-61</Citation>
<ArticleIdList><ArticleId IdType="pubmed">11724861</ArticleId>
</ArticleIdList>
</Reference>
<Reference><Citation>J Clin Microbiol. 2008 Nov;46(11):3653-9</Citation>
<ArticleIdList><ArticleId IdType="pubmed">18799705</ArticleId>
</ArticleIdList>
</Reference>
<Reference><Citation>J Virol Methods. 2013 Jan;187(1):177-81</Citation>
<ArticleIdList><ArticleId IdType="pubmed">23022325</ArticleId>
</ArticleIdList>
</Reference>
<Reference><Citation>Virol J. 2008 Jul 18;5:82</Citation>
<ArticleIdList><ArticleId IdType="pubmed">18638365</ArticleId>
</ArticleIdList>
</Reference>
<Reference><Citation>J Virol Methods. 2012 Aug;183(2):99-105</Citation>
<ArticleIdList><ArticleId IdType="pubmed">22449758</ArticleId>
</ArticleIdList>
</Reference>
<Reference><Citation>S Afr Med J. 1980 Nov 15;58(20):803-6</Citation>
<ArticleIdList><ArticleId IdType="pubmed">7192434</ArticleId>
</ArticleIdList>
</Reference>
<Reference><Citation>Drug Discov Today. 2009 Jan;14(1-2):16-24</Citation>
<ArticleIdList><ArticleId IdType="pubmed">18983932</ArticleId>
</ArticleIdList>
</Reference>
<Reference><Citation>Rev Sci Tech. 1993 Jun;12(2):435-50</Citation>
<ArticleIdList><ArticleId IdType="pubmed">8400384</ArticleId>
</ArticleIdList>
</Reference>
<Reference><Citation>J Virol Methods. 2016 Oct;236:170-177</Citation>
<ArticleIdList><ArticleId IdType="pubmed">27474493</ArticleId>
</ArticleIdList>
</Reference>
<Reference><Citation>Vaccine. 2010 Mar 8;28(11):2330-9</Citation>
<ArticleIdList><ArticleId IdType="pubmed">20056185</ArticleId>
</ArticleIdList>
</Reference>
<Reference><Citation>J Virol Methods. 2012 Feb;179(2):373-82</Citation>
<ArticleIdList><ArticleId IdType="pubmed">22172972</ArticleId>
</ArticleIdList>
</Reference>
<Reference><Citation>Vet Microbiol. 2007 Mar 31;121(1-2):29-38</Citation>
<ArticleIdList><ArticleId IdType="pubmed">17187944</ArticleId>
</ArticleIdList>
</Reference>
<Reference><Citation>J Clin Microbiol. 1984 Feb;19(2):225-9</Citation>
<ArticleIdList><ArticleId IdType="pubmed">6538206</ArticleId>
</ArticleIdList>
</Reference>
<Reference><Citation>Curr Protoc Protein Sci. 2013 Sep 24;73:9.9.1-9.9.23</Citation>
<ArticleIdList><ArticleId IdType="pubmed">24510596</ArticleId>
</ArticleIdList>
</Reference>
<Reference><Citation>J Virol Methods. 2006 Jan;131(1):34-40</Citation>
<ArticleIdList><ArticleId IdType="pubmed">16102851</ArticleId>
</ArticleIdList>
</Reference>
<Reference><Citation>Plant Biotechnol J. 2010 Jun;8(5):620-37</Citation>
<ArticleIdList><ArticleId IdType="pubmed">20233333</ArticleId>
</ArticleIdList>
</Reference>
<Reference><Citation>J Virol Methods. 2007 Mar;140(1-2):106-14</Citation>
<ArticleIdList><ArticleId IdType="pubmed">17174410</ArticleId>
</ArticleIdList>
</Reference>
<Reference><Citation>Lancet Glob Health. 2016 Nov;4(11):e864-e871</Citation>
<ArticleIdList><ArticleId IdType="pubmed">27692776</ArticleId>
</ArticleIdList>
</Reference>
<Reference><Citation>MAbs. 2016 Nov/Dec;8(8):1456-1466</Citation>
<ArticleIdList><ArticleId IdType="pubmed">27559626</ArticleId>
</ArticleIdList>
</Reference>
<Reference><Citation>Onderstepoort J Vet Res. 2014 May 27;81(1):</Citation>
<ArticleIdList><ArticleId IdType="pubmed">25686466</ArticleId>
</ArticleIdList>
</Reference>
<Reference><Citation>Trop Anim Health Prod. 2014 Apr;46(4):629-34</Citation>
<ArticleIdList><ArticleId IdType="pubmed">24464589</ArticleId>
</ArticleIdList>
</Reference>
<Reference><Citation>J Virol. 2007 May;81(10):5246-56</Citation>
<ArticleIdList><ArticleId IdType="pubmed">17329326</ArticleId>
</ArticleIdList>
</Reference>
<Reference><Citation>Virol J. 2009 Aug 13;6:125</Citation>
<ArticleIdList><ArticleId IdType="pubmed">19678951</ArticleId>
</ArticleIdList>
</Reference>
<Reference><Citation>J Virol Methods. 2003 Nov;113(2):103-12</Citation>
<ArticleIdList><ArticleId IdType="pubmed">14553896</ArticleId>
</ArticleIdList>
</Reference>
<Reference><Citation>PLoS One. 2013 Aug 12;8(8):e71842</Citation>
<ArticleIdList><ArticleId IdType="pubmed">23951256</ArticleId>
</ArticleIdList>
</Reference>
<Reference><Citation>J Virol Methods. 2012 Mar;180(1-2):68-74</Citation>
<ArticleIdList><ArticleId IdType="pubmed">22240075</ArticleId>
</ArticleIdList>
</Reference>
<Reference><Citation>J Virol Methods. 2011 Nov;177(2):140-6</Citation>
<ArticleIdList><ArticleId IdType="pubmed">21827790</ArticleId>
</ArticleIdList>
</Reference>
<Reference><Citation>Int J Mol Sci. 2011;12(11):8275-87</Citation>
<ArticleIdList><ArticleId IdType="pubmed">22174663</ArticleId>
</ArticleIdList>
</Reference>
<Reference><Citation>J Clin Microbiol. 2009 Mar;47(3):645-51</Citation>
<ArticleIdList><ArticleId IdType="pubmed">19109471</ArticleId>
</ArticleIdList>
</Reference>
<Reference><Citation>J Virol Methods. 2005 Mar;124(1-2):173-81</Citation>
<ArticleIdList><ArticleId IdType="pubmed">15664066</ArticleId>
</ArticleIdList>
</Reference>
<Reference><Citation>Plant Biotechnol J. 2009 Sep;7(7):682-93</Citation>
<ArticleIdList><ArticleId IdType="pubmed">19627561</ArticleId>
</ArticleIdList>
</Reference>
<Reference><Citation>J Clin Virol. 2012 Aug;54(4):308-12</Citation>
<ArticleIdList><ArticleId IdType="pubmed">22683006</ArticleId>
</ArticleIdList>
</Reference>
<Reference><Citation>Vet Res. 2010 Nov-Dec;41(6):61</Citation>
<ArticleIdList><ArticleId IdType="pubmed">21188836</ArticleId>
</ArticleIdList>
</Reference>
<Reference><Citation>Plant Biotechnol J. 2010 Jan;8(1):38-46</Citation>
<ArticleIdList><ArticleId IdType="pubmed">19929900</ArticleId>
</ArticleIdList>
</Reference>
<Reference><Citation>J Gen Virol. 2007 May;88(Pt 5):1460-9</Citation>
<ArticleIdList><ArticleId IdType="pubmed">17412974</ArticleId>
</ArticleIdList>
</Reference>
<Reference><Citation>Emerg Infect Dis. 2013 Dec;19(12):</Citation>
<ArticleIdList><ArticleId IdType="pubmed">29360021</ArticleId>
</ArticleIdList>
</Reference>
<Reference><Citation>J Clin Microbiol. 2002 May;40(5):1587-91</Citation>
<ArticleIdList><ArticleId IdType="pubmed">11980926</ArticleId>
</ArticleIdList>
</Reference>
<Reference><Citation>J Clin Microbiol. 2002 Jul;40(7):2323-30</Citation>
<ArticleIdList><ArticleId IdType="pubmed">12089242</ArticleId>
</ArticleIdList>
</Reference>
<Reference><Citation>Hum Vaccin Immunother. 2017 Dec 2;13(12):2912-2917</Citation>
<ArticleIdList><ArticleId IdType="pubmed">28846485</ArticleId>
</ArticleIdList>
</Reference>
<Reference><Citation>Curr Opin Biotechnol. 2015 Apr;32:156-162</Citation>
<ArticleIdList><ArticleId IdType="pubmed">25562816</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="Mbewana, Sandiswa" sort="Mbewana, Sandiswa" uniqKey="Mbewana S" first="Sandiswa" last="Mbewana">Sandiswa Mbewana</name>
</noRegion>
<name sortKey="Ferreira, Maryke L" sort="Ferreira, Maryke L" uniqKey="Ferreira M" first="Maryke L" last="Ferreira">Maryke L. Ferreira</name>
<name sortKey="Majiwa, Phelix A O" sort="Majiwa, Phelix A O" uniqKey="Majiwa P" first="Phelix A O" last="Majiwa">Phelix A O. Majiwa</name>
<name sortKey="Mareledwane, Vuyokazi" sort="Mareledwane, Vuyokazi" uniqKey="Mareledwane V" first="Vuyokazi" last="Mareledwane">Vuyokazi Mareledwane</name>
<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>
<name sortKey="Rybicki, Edward P" sort="Rybicki, Edward P" uniqKey="Rybicki E" first="Edward P" last="Rybicki">Edward P. Rybicki</name>
<name sortKey="Weber, Brandon" sort="Weber, Brandon" uniqKey="Weber B" first="Brandon" last="Weber">Brandon Weber</name>
</country>
</tree>
</affiliations>
</record>

Pour manipuler ce document sous Unix (Dilib)

EXPLOR_STEP=$WICRI_ROOT/Bois/explor/AgrobacTransV1/Data/Main/Exploration

HfdSelect -h $EXPLOR_STEP/biblio.hfd -nk 000181 | SxmlIndent | more

HfdSelect -h $EXPLOR_AREA/Data/Main/Exploration/biblio.hfd -nk 000181 | SxmlIndent | more

Pour mettre un lien sur cette page dans le réseau Wicri

{{Explor lien
   |wiki=    Bois
   |area=    AgrobacTransV1
   |flux=    Main
   |étape=   Exploration
   |type=    RBID
   |clé=     pubmed:30537953
   |texte=   Expression of Rift Valley fever virus N-protein in Nicotiana benthamiana for use as a diagnostic antigen.
}}

Pour générer des pages wiki

HfdIndexSelect -h $EXPLOR_AREA/Data/Main/Exploration/RBID.i   -Sk "pubmed:30537953" \
       | HfdSelect -Kh $EXPLOR_AREA/Data/Main/Exploration/biblio.hfd   \
       | NlmPubMed2Wicri -a AgrobacTransV1

This area was generated with Dilib version V0.6.38.
Data generation: Fri Nov 20 15:45:55 2020. Site generation: Wed Mar 6 15:24:41 2024

	Serveur d'exploration sur l'agrobacterium et la transgénèse
	Attention, ce site est en cours de développement ! Attention, site généré par des moyens informatiques à partir de corpus bruts. Les informations ne sont donc pas validées.

Serveur d'exploration sur l'agrobacterium et la transgénèse

Expression of Rift Valley fever virus N-protein in Nicotiana benthamiana for use as a diagnostic antigen.

Expression of Rift Valley fever virus N-protein in Nicotiana benthamiana for use as a diagnostic antigen.

Source :

Descripteurs français

English descriptors

Abstract

Links toward previous steps (curation, corpus...)

Le document en format XML

Pour manipuler ce document sous Unix (Dilib)

Pour mettre un lien sur cette page dans le réseau Wicri

Pour générer des pages wiki