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Pseudotyped vesicular stomatitis virus for analysis of virus entry mediated by SARS coronavirus spike proteins.
Methods Mol Biol. 2008; 454:331-8.MM

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.

Authors+Show Affiliations

Department of Virology I, National Institute of Infectious Diseases, Musashimurayama, Tokyo, Japan.No affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

19057867

Citation

Fukushi, Shuetsu, et al. "Pseudotyped Vesicular Stomatitis Virus for Analysis of Virus Entry Mediated By SARS Coronavirus Spike Proteins." Methods in Molecular Biology (Clifton, N.J.), vol. 454, 2008, pp. 331-8.
Fukushi S, Watanabe R, Taguchi F. Pseudotyped vesicular stomatitis virus for analysis of virus entry mediated by SARS coronavirus spike proteins. Methods Mol Biol. 2008;454:331-8.
Fukushi, S., Watanabe, R., & Taguchi, F. (2008). Pseudotyped vesicular stomatitis virus for analysis of virus entry mediated by SARS coronavirus spike proteins. Methods in Molecular Biology (Clifton, N.J.), 454, 331-8. https://doi.org/10.1007/978-1-59745-181-9_23
Fukushi S, Watanabe R, Taguchi F. Pseudotyped Vesicular Stomatitis Virus for Analysis of Virus Entry Mediated By SARS Coronavirus Spike Proteins. Methods Mol Biol. 2008;454:331-8. PubMed PMID: 19057867.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Pseudotyped vesicular stomatitis virus for analysis of virus entry mediated by SARS coronavirus spike proteins. AU - Fukushi,Shuetsu, AU - Watanabe,Rie, AU - Taguchi,Fumihiro, PY - 2008/12/6/pubmed PY - 2009/3/20/medline PY - 2008/12/6/entrez SP - 331 EP - 8 JF - Methods in molecular biology (Clifton, N.J.) JO - Methods Mol Biol VL - 454 N2 - 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. SN - 1064-3745 UR - https://www.unboundmedicine.com/medline/citation/19057867/Pseudotyped_vesicular_stomatitis_virus_for_analysis_of_virus_entry_mediated_by_SARS_coronavirus_spike_proteins_ L2 - https://dx.doi.org/10.1007/978-1-59745-181-9_23 DB - PRIME DP - Unbound Medicine ER -