SrasV1, PubMed, Checkpoint, bibRecord, 001A56

Pseudotyped vesicular stomatitis virus for analysis of virus entry mediated by SARS coronavirus spike proteins.

Identifieur interne : 001A56 ( PubMed/Checkpoint ); précédent : 001A55; suivant : 001A57

Pseudotyped vesicular stomatitis virus for analysis of virus entry mediated by SARS coronavirus spike proteins.

Auteurs : Shuetsu Fukushi [Japon] ; Rie Watanabe ; Fumihiro Taguchi

Source :

Methods in molecular biology (Clifton, N.J.) [ 1064-3745 ] ; 2008.

RBID : pubmed:19057867

Descripteurs français

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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<term>Virus de la stomatite vésiculeuse de type Indiana (métabolisme)</term>
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<term>Virus du SRAS (physiologie)</term>
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<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>
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Pseudotyped vesicular stomatitis virus for analysis of virus entry mediated by SARS coronavirus spike proteins.

Pseudotyped vesicular stomatitis virus for analysis of virus entry mediated by SARS coronavirus spike proteins.

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