Pseudotyped vesicular stomatitis virus for analysis of virus entry mediated by SARS coronavirus spike proteins.
Identifieur interne : 003050 ( Main/Exploration ); précédent : 003049; suivant : 003051Pseudotyped vesicular stomatitis virus for analysis of virus entry mediated by SARS coronavirus spike proteins.
Auteurs : Shuetsu Fukushi [Japon] ; Rie Watanabe ; Fumihiro TaguchiSource :
- Methods in molecular biology (Clifton, N.J.) [ 1064-3745 ] ; 2008.
Descripteurs français
- KwdFr :
- Animaux, Cellules Vero, Glycoprotéine de spicule des coronavirus, Glycoprotéines membranaires (métabolisme), Humains, Lignée cellulaire, Microscopie de fluorescence, Modèles biologiques, Protéines de l'enveloppe virale (métabolisme), Protéines à fluorescence verte (génétique), Protéines à fluorescence verte (métabolisme), Pénétration virale, Virus de la stomatite vésiculeuse de type Indiana (génétique), Virus de la stomatite vésiculeuse de type Indiana (métabolisme), Virus de la stomatite vésiculeuse de type Indiana (physiologie), Virus du SRAS (métabolisme), Virus du SRAS (physiologie).
- MESH :
- génétique : Protéines à fluorescence verte, Virus de la stomatite vésiculeuse de type Indiana.
- métabolisme : Glycoprotéines membranaires, Protéines de l'enveloppe virale, Protéines à fluorescence verte, Virus de la stomatite vésiculeuse de type Indiana, Virus du SRAS.
- physiologie : Virus de la stomatite vésiculeuse de type Indiana, Virus du SRAS.
- Animaux, Cellules Vero, Glycoprotéine de spicule des coronavirus, Humains, Lignée cellulaire, Microscopie de fluorescence, Modèles biologiques, Pénétration virale.
English descriptors
- KwdEn :
- Animals, Cell Line, Chlorocebus aethiops, Green Fluorescent Proteins (genetics), Green Fluorescent Proteins (metabolism), Humans, Membrane Glycoproteins (metabolism), Microscopy, Fluorescence, Models, Biological, SARS Virus (metabolism), SARS Virus (physiology), Spike Glycoprotein, Coronavirus, Vero Cells, Vesicular stomatitis Indiana virus (genetics), Vesicular stomatitis Indiana virus (metabolism), Vesicular stomatitis Indiana virus (physiology), Viral Envelope Proteins (metabolism), Virus Internalization.
- MESH :
- chemical , genetics : Green Fluorescent Proteins.
- chemical , metabolism : Green Fluorescent Proteins, Membrane Glycoproteins, Viral Envelope Proteins.
- genetics : Vesicular stomatitis Indiana virus.
- metabolism : SARS Virus, Vesicular stomatitis Indiana virus.
- physiology : SARS Virus, Vesicular stomatitis Indiana virus.
- Animals, Cell Line, Chlorocebus aethiops, Humans, Microscopy, Fluorescence, Models, Biological, Spike Glycoprotein, Coronavirus, Vero Cells, Virus Internalization.
Abstract
Severe acute respiratory syndrome (SARS) coronavirus (CoV) contains a spike (S) protein that binds to a receptor molecule (angiotensin-converting enzyme 2; ACE2), induces membrane fusion, and serves as a neutralizing epitope. To study the functions of the S protein, we describe here the generation of SARS-CoV S protein-bearing vesicular stomatitis virus (VSV) pseudotype using a VSVdeltaG*/GFP system in which the G gene is replaced by the green fluorescent protein (GFP) gene (VSV-SARS-CoV-St19/GFP). Partial deletion of the cytoplasmic domain of SARS-CoV S protein (SARS-CoV-St19) allowed efficient incorporation into the VSV particle that enabled the generation of a high titer of pseudotype virus. Neutralization assay with anti-SARS-CoV antibody revealed that VSV-SARS-St19/GFP pseudotype infection is mediated by SARS-CoV S protein. The VSVdeltaaG*/SEAP system, which secretes alkaline phosphatase instead of GFP, was also generated as a VSV pseudotype having SARS-CoV S protein (VSV-SARS-CoV-St19/SEAP). This system enabled high-throughput analysis of SARS-CoV S protein-mediated cell entry by measuring alkaline phosphatase activity. Thus, VSV pseudotyped with SARS-CoV S protein is useful for developing a rapid detection system for neutralizing antibody specific for SARS-CoV infection as well as studying the S-mediated cell entry of SARS-CoV.
DOI: 10.1007/978-1-59745-181-9_23
PubMed: 19057867
Affiliations:
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Le document en format XML
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uniqKey="Watanabe R" first="Rie" last="Watanabe">Rie Watanabe</name></author><author><name sortKey="Taguchi, Fumihiro" sort="Taguchi, Fumihiro" uniqKey="Taguchi F" first="Fumihiro" last="Taguchi">Fumihiro Taguchi</name></author></analytic><series><title level="j">Methods in molecular biology (Clifton, N.J.)</title><idno type="ISSN">1064-3745</idno><imprint><date when="2008" type="published">2008</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Animals</term><term>Cell Line</term><term>Chlorocebus aethiops</term><term>Green Fluorescent Proteins (genetics)</term><term>Green Fluorescent Proteins (metabolism)</term><term>Humans</term><term>Membrane Glycoproteins (metabolism)</term><term>Microscopy, Fluorescence</term><term>Models, Biological</term><term>SARS Virus (metabolism)</term><term>SARS Virus (physiology)</term><term>Spike Glycoprotein, Coronavirus</term><term>Vero Cells</term><term>Vesicular stomatitis Indiana virus (genetics)</term><term>Vesicular stomatitis Indiana virus (metabolism)</term><term>Vesicular stomatitis Indiana virus (physiology)</term><term>Viral Envelope Proteins (metabolism)</term><term>Virus Internalization</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Animaux</term><term>Cellules Vero</term><term>Glycoprotéine de spicule des coronavirus</term><term>Glycoprotéines membranaires (métabolisme)</term><term>Humains</term><term>Lignée cellulaire</term><term>Microscopie de fluorescence</term><term>Modèles biologiques</term><term>Protéines de l'enveloppe virale (métabolisme)</term><term>Protéines à fluorescence verte (génétique)</term><term>Protéines à fluorescence verte (métabolisme)</term><term>Pénétration virale</term><term>Virus de la stomatite vésiculeuse de type Indiana (génétique)</term><term>Virus de la stomatite vésiculeuse de type Indiana (métabolisme)</term><term>Virus de la stomatite vésiculeuse de type Indiana (physiologie)</term><term>Virus du SRAS (métabolisme)</term><term>Virus du SRAS (physiologie)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>Green Fluorescent Proteins</term></keywords><keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>Green Fluorescent Proteins</term><term>Membrane Glycoproteins</term><term>Viral Envelope Proteins</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Vesicular stomatitis Indiana virus</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>Protéines à fluorescence verte</term><term>Virus de la stomatite vésiculeuse de type Indiana</term></keywords><keywords scheme="MESH" qualifier="metabolism" xml:lang="en"><term>SARS Virus</term><term>Vesicular stomatitis Indiana virus</term></keywords><keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>Glycoprotéines membranaires</term><term>Protéines de 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fluorescence</term><term>Modèles biologiques</term><term>Pénétration virale</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Severe acute respiratory syndrome (SARS) coronavirus (CoV) contains a spike (S) protein that binds to a receptor molecule (angiotensin-converting enzyme 2; ACE2), induces membrane fusion, and serves as a neutralizing epitope. To study the functions of the S protein, we describe here the generation of SARS-CoV S protein-bearing vesicular stomatitis virus (VSV) pseudotype using a VSVdeltaG*/GFP system in which the G gene is replaced by the green fluorescent protein (GFP) gene (VSV-SARS-CoV-St19/GFP). Partial deletion of the cytoplasmic domain of SARS-CoV S protein (SARS-CoV-St19) allowed efficient incorporation into the VSV particle that enabled the generation of a high titer of pseudotype virus. Neutralization assay with anti-SARS-CoV antibody revealed that VSV-SARS-St19/GFP pseudotype infection is mediated by SARS-CoV S protein. The VSVdeltaaG*/SEAP system, which secretes alkaline phosphatase instead of GFP, was also generated as a VSV pseudotype having SARS-CoV S protein (VSV-SARS-CoV-St19/SEAP). This system enabled high-throughput analysis of SARS-CoV S protein-mediated cell entry by measuring alkaline phosphatase activity. Thus, VSV pseudotyped with SARS-CoV S protein is useful for developing a rapid detection system for neutralizing antibody specific for SARS-CoV infection as well as studying the S-mediated cell entry of SARS-CoV.</div></front></TEI><affiliations><list><country><li>Japon</li></country><region><li>Région de Kantō</li></region><settlement><li>Tokyo</li></settlement></list><tree><noCountry><name sortKey="Taguchi, Fumihiro" sort="Taguchi, Fumihiro" uniqKey="Taguchi F" first="Fumihiro" last="Taguchi">Fumihiro Taguchi</name><name sortKey="Watanabe, Rie" sort="Watanabe, Rie" uniqKey="Watanabe R" first="Rie" last="Watanabe">Rie Watanabe</name></noCountry><country name="Japon"><region name="Région de Kantō"><name sortKey="Fukushi, Shuetsu" sort="Fukushi, Shuetsu" uniqKey="Fukushi S" first="Shuetsu" last="Fukushi">Shuetsu Fukushi</name></region></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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