Construction of a non-infectious SARS coronavirus replicon for application in drug screening and analysis of viral protein function.
Identifieur interne : 001B02 ( PubMed/Corpus ); précédent : 001B01; suivant : 001B03Construction of a non-infectious SARS coronavirus replicon for application in drug screening and analysis of viral protein function.
Auteurs : Jian-Min Wang ; Lin-Fa Wang ; Zheng-Li ShiSource :
- Biochemical and biophysical research communications [ 1090-2104 ] ; 2008.
English descriptors
- KwdEn :
- Antiviral Agents (pharmacology), Drug Evaluation, Preclinical (methods), Gene Deletion, Green Fluorescent Proteins (analysis), Green Fluorescent Proteins (genetics), Humans, RNA Replicase (genetics), RNA Replicase (metabolism), Replicon, SARS Virus (drug effects), SARS Virus (genetics), SARS Virus (metabolism), Viral Nonstructural Proteins (genetics), Viral Nonstructural Proteins (metabolism), Virus Replication (drug effects), Virus Replication (genetics).
- MESH :
- chemical , analysis : Green Fluorescent Proteins.
- chemical , genetics : Green Fluorescent Proteins, RNA Replicase, Viral Nonstructural Proteins.
- chemical , metabolism : RNA Replicase, Viral Nonstructural Proteins.
- chemical , pharmacology : Antiviral Agents.
- drug effects : SARS Virus, Virus Replication.
- genetics : SARS Virus, Virus Replication.
- metabolism : SARS Virus.
- methods : Drug Evaluation, Preclinical.
- Gene Deletion, Humans, Replicon.
Abstract
Severe acute respiratory syndrome virus (SARS-CoV) was the causative agent of the SARS outbreaks in 2002-2003. A safer in vitro system is desirable for conducting research on SARS-CoV and to screen for antiviral drugs against the virus. Based on the infectious cDNA clone of rSARS-CoV-DeltaE, in which the E gene has been deleted, a safe non-infectious replicon was constructed by replacing the S gene with the enhanced green fluorescent protein (eGFP) gene. Successful replication was achieved as evident from continuous expression of eGFP detected by both fluorescence and Western blot. Treatment with antiviral drugs demonstrated that the replication could be significantly inhibited by 0.4 mg/ml of cysteine proteinase inhibitor E-64D, but not by ribavirin. The same replicons containing further deletion of the coding regions for non-structural proteins (nsp) 1, 2 or 16 confirmed previous observation that nsp16, but not nsp1 or nsp2, was essential for efficient viral replication or transcription.
DOI: 10.1016/j.bbrc.2008.06.129
PubMed: 18619943
Links to Exploration step
pubmed:18619943Le document en format XML
<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Construction of a non-infectious SARS coronavirus replicon for application in drug screening and analysis of viral protein function.</title><author><name sortKey="Wang, Jian Min" sort="Wang, Jian Min" uniqKey="Wang J" first="Jian-Min" last="Wang">Jian-Min Wang</name><affiliation><nlm:affiliation>State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, 44 Xiao Hong Shan, Wuhan, Hubei 430071, PR China.</nlm:affiliation></affiliation></author><author><name sortKey="Wang, Lin Fa" sort="Wang, Lin Fa" uniqKey="Wang L" first="Lin-Fa" last="Wang">Lin-Fa Wang</name></author><author><name sortKey="Shi, Zheng Li" sort="Shi, Zheng Li" uniqKey="Shi Z" first="Zheng-Li" last="Shi">Zheng-Li Shi</name></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="2008">2008</date><idno type="RBID">pubmed:18619943</idno><idno type="pmid">18619943</idno><idno type="doi">10.1016/j.bbrc.2008.06.129</idno><idno type="wicri:Area/PubMed/Corpus">001B02</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Corpus" wicri:corpus="PubMed">001B02</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en">Construction of a non-infectious SARS coronavirus replicon for application in drug screening and analysis of viral protein function.</title><author><name sortKey="Wang, Jian Min" sort="Wang, Jian Min" uniqKey="Wang J" first="Jian-Min" last="Wang">Jian-Min Wang</name><affiliation><nlm:affiliation>State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, 44 Xiao Hong Shan, Wuhan, Hubei 430071, PR China.</nlm:affiliation></affiliation></author><author><name sortKey="Wang, Lin Fa" sort="Wang, Lin Fa" uniqKey="Wang L" first="Lin-Fa" last="Wang">Lin-Fa Wang</name></author><author><name sortKey="Shi, Zheng Li" sort="Shi, Zheng Li" uniqKey="Shi Z" first="Zheng-Li" last="Shi">Zheng-Li Shi</name></author></analytic><series><title level="j">Biochemical and biophysical research communications</title><idno type="eISSN">1090-2104</idno><imprint><date when="2008" type="published">2008</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Antiviral Agents (pharmacology)</term><term>Drug Evaluation, Preclinical (methods)</term><term>Gene Deletion</term><term>Green Fluorescent Proteins (analysis)</term><term>Green Fluorescent Proteins (genetics)</term><term>Humans</term><term>RNA Replicase (genetics)</term><term>RNA Replicase (metabolism)</term><term>Replicon</term><term>SARS Virus (drug effects)</term><term>SARS Virus (genetics)</term><term>SARS Virus (metabolism)</term><term>Viral Nonstructural Proteins (genetics)</term><term>Viral Nonstructural Proteins (metabolism)</term><term>Virus Replication (drug effects)</term><term>Virus Replication (genetics)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="analysis" xml:lang="en"><term>Green Fluorescent Proteins</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>Green Fluorescent Proteins</term><term>RNA Replicase</term><term>Viral Nonstructural Proteins</term></keywords><keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>RNA Replicase</term><term>Viral Nonstructural Proteins</term></keywords><keywords scheme="MESH" type="chemical" qualifier="pharmacology" xml:lang="en"><term>Antiviral Agents</term></keywords><keywords scheme="MESH" qualifier="drug effects" xml:lang="en"><term>SARS Virus</term><term>Virus Replication</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>SARS Virus</term><term>Virus Replication</term></keywords><keywords scheme="MESH" qualifier="metabolism" xml:lang="en"><term>SARS Virus</term></keywords><keywords scheme="MESH" qualifier="methods" xml:lang="en"><term>Drug Evaluation, Preclinical</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Gene Deletion</term><term>Humans</term><term>Replicon</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Severe acute respiratory syndrome virus (SARS-CoV) was the causative agent of the SARS outbreaks in 2002-2003. A safer in vitro system is desirable for conducting research on SARS-CoV and to screen for antiviral drugs against the virus. Based on the infectious cDNA clone of rSARS-CoV-DeltaE, in which the E gene has been deleted, a safe non-infectious replicon was constructed by replacing the S gene with the enhanced green fluorescent protein (eGFP) gene. Successful replication was achieved as evident from continuous expression of eGFP detected by both fluorescence and Western blot. Treatment with antiviral drugs demonstrated that the replication could be significantly inhibited by 0.4 mg/ml of cysteine proteinase inhibitor E-64D, but not by ribavirin. The same replicons containing further deletion of the coding regions for non-structural proteins (nsp) 1, 2 or 16 confirmed previous observation that nsp16, but not nsp1 or nsp2, was essential for efficient viral replication or transcription.</div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">18619943</PMID><DateCompleted><Year>2008</Year><Month>08</Month><Day>25</Day></DateCompleted><DateRevised><Year>2020</Year><Month>03</Month><Day>31</Day></DateRevised><Article PubModel="Print-Electronic"><Journal><ISSN IssnType="Electronic">1090-2104</ISSN><JournalIssue CitedMedium="Internet"><Volume>374</Volume><Issue>1</Issue><PubDate><Year>2008</Year><Month>Sep</Month><Day>12</Day></PubDate></JournalIssue><Title>Biochemical and biophysical research communications</Title><ISOAbbreviation>Biochem. Biophys. Res. Commun.</ISOAbbreviation></Journal><ArticleTitle>Construction of a non-infectious SARS coronavirus replicon for application in drug screening and analysis of viral protein function.</ArticleTitle><Pagination><MedlinePgn>138-42</MedlinePgn></Pagination><ELocationID EIdType="doi" ValidYN="Y">10.1016/j.bbrc.2008.06.129</ELocationID><Abstract><AbstractText>Severe acute respiratory syndrome virus (SARS-CoV) was the causative agent of the SARS outbreaks in 2002-2003. A safer in vitro system is desirable for conducting research on SARS-CoV and to screen for antiviral drugs against the virus. Based on the infectious cDNA clone of rSARS-CoV-DeltaE, in which the E gene has been deleted, a safe non-infectious replicon was constructed by replacing the S gene with the enhanced green fluorescent protein (eGFP) gene. Successful replication was achieved as evident from continuous expression of eGFP detected by both fluorescence and Western blot. Treatment with antiviral drugs demonstrated that the replication could be significantly inhibited by 0.4 mg/ml of cysteine proteinase inhibitor E-64D, but not by ribavirin. The same replicons containing further deletion of the coding regions for non-structural proteins (nsp) 1, 2 or 16 confirmed previous observation that nsp16, but not nsp1 or nsp2, was essential for efficient viral replication or transcription.</AbstractText></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Wang</LastName><ForeName>Jian-Min</ForeName><Initials>JM</Initials><AffiliationInfo><Affiliation>State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, 44 Xiao Hong Shan, Wuhan, Hubei 430071, PR China.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Wang</LastName><ForeName>Lin-Fa</ForeName><Initials>LF</Initials></Author><Author ValidYN="Y"><LastName>Shi</LastName><ForeName>Zheng-Li</ForeName><Initials>ZL</Initials></Author></AuthorList><Language>eng</Language><PublicationTypeList><PublicationType UI="D016428">Journal Article</PublicationType><PublicationType UI="D013485">Research Support, Non-U.S. 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