Construction of a non-infectious SARS coronavirus replicon for application in drug screening and analysis of viral protein function
Identifieur interne : 003145 ( Main/Exploration ); précédent : 003144; suivant : 003146Construction of a non-infectious SARS coronavirus replicon for application in drug screening and analysis of viral protein function
Auteurs : Jian-Min Wang [République populaire de Chine] ; Lin-Fa Wang [Australie] ; Zheng-Li Shi [République populaire de Chine]Source :
- Biochemical and Biophysical Research Communications [ 0006-291X ] ; 2008.
Descripteurs français
- KwdFr :
- Antiviraux (pharmacologie), Délétion de gène, Humains, Protéines virales non structurales (génétique), Protéines virales non structurales (métabolisme), Protéines à fluorescence verte (analyse), Protéines à fluorescence verte (génétique), RNA replicase (génétique), RNA replicase (métabolisme), Réplication virale (), Réplication virale (génétique), Réplicon, Virus du SRAS (), Virus du SRAS (génétique), Virus du SRAS (métabolisme), Évaluation préclinique de médicament ().
- MESH :
- analyse : Protéines à fluorescence verte.
- génétique : Protéines virales non structurales, Protéines à fluorescence verte, RNA replicase, Réplication virale, Virus du SRAS.
- métabolisme : Protéines virales non structurales, RNA replicase, Virus du SRAS.
- pharmacologie : Antiviraux.
- Délétion de gène, Humains, Réplication virale, Réplicon, Virus du SRAS, Évaluation préclinique de médicament.
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-ΔE, 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.
Url:
DOI: 10.1016/j.bbrc.2008.06.129
PubMed: 18619943
PubMed Central: 7092913
Affiliations:
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Le document en format XML
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sortKey="Wang, Jian Min" sort="Wang, Jian Min" uniqKey="Wang J" first="Jian-Min" last="Wang">Jian-Min Wang</name><affiliation wicri:level="1"><nlm:aff id="aff1">State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, 44 Xiao Hong Shan, Wuhan, Hubei 430071, PR China</nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, 44 Xiao Hong Shan, Wuhan, Hubei 430071</wicri:regionArea><wicri:noRegion>Hubei 430071</wicri:noRegion></affiliation></author><author><name sortKey="Wang, Lin Fa" sort="Wang, Lin Fa" uniqKey="Wang L" first="Lin-Fa" last="Wang">Lin-Fa Wang</name><affiliation wicri:level="1"><nlm:aff id="aff2">CSIRO Australian Animal Health Laboratory and Australian Biosecurity Cooperative Research Center for Emerging Infectious Diseases, Geelong, Australia</nlm:aff><country xml:lang="fr">Australie</country><wicri:regionArea>CSIRO Australian Animal Health Laboratory and Australian Biosecurity Cooperative Research Center for Emerging Infectious Diseases, Geelong</wicri:regionArea><wicri:noRegion>Geelong</wicri:noRegion></affiliation></author><author><name sortKey="Shi, Zheng Li" sort="Shi, Zheng Li" uniqKey="Shi Z" first="Zheng-Li" last="Shi">Zheng-Li Shi</name><affiliation wicri:level="1"><nlm:aff id="aff1">State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, 44 Xiao Hong Shan, Wuhan, Hubei 430071, PR China</nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, 44 Xiao Hong Shan, Wuhan, Hubei 430071</wicri:regionArea><wicri:noRegion>Hubei 430071</wicri:noRegion></affiliation></author></analytic><series><title level="j">Biochemical and Biophysical Research Communications</title><idno type="ISSN">0006-291X</idno><idno type="eISSN">1090-2104</idno><imprint><date when="2008">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="KwdFr" xml:lang="fr"><term>Antiviraux (pharmacologie)</term><term>Délétion de gène</term><term>Humains</term><term>Protéines virales non structurales (génétique)</term><term>Protéines virales non structurales (métabolisme)</term><term>Protéines à fluorescence verte (analyse)</term><term>Protéines à fluorescence verte (génétique)</term><term>RNA replicase (génétique)</term><term>RNA replicase (métabolisme)</term><term>Réplication virale ()</term><term>Réplication virale (génétique)</term><term>Réplicon</term><term>Virus du SRAS ()</term><term>Virus du SRAS (génétique)</term><term>Virus du SRAS (métabolisme)</term><term>Évaluation préclinique de médicament ()</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="analyse" xml:lang="fr"><term>Protéines à fluorescence verte</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="génétique" xml:lang="fr"><term>Protéines virales non structurales</term><term>Protéines à fluorescence verte</term><term>RNA replicase</term><term>Réplication virale</term><term>Virus du SRAS</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" qualifier="métabolisme" xml:lang="fr"><term>Protéines virales non structurales</term><term>RNA replicase</term><term>Virus du SRAS</term></keywords><keywords scheme="MESH" qualifier="pharmacologie" xml:lang="fr"><term>Antiviraux</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Gene Deletion</term><term>Humans</term><term>Replicon</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Délétion de gène</term><term>Humains</term><term>Réplication virale</term><term>Réplicon</term><term>Virus du SRAS</term><term>Évaluation préclinique de médicament</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p>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-ΔE, 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.</p></div></front><back><div1 type="bibliography"><listBibl><biblStruct><analytic><author><name sortKey="Peiris, J S" uniqKey="Peiris J">J.S. Peiris</name></author><author><name sortKey="Yuen, K Y" uniqKey="Yuen K">K.Y. Yuen</name></author><author><name sortKey="Osterhaus, A D" uniqKey="Osterhaus A">A.D. Osterhaus</name></author><author><name sortKey="Stohr, K" uniqKey="Stohr K">K. Stohr</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Chan Yeung, M" uniqKey="Chan Yeung M">M. Chan-Yeung</name></author><author><name sortKey="Xu, R H" uniqKey="Xu R">R.H. Xu</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Lau, S K" uniqKey="Lau S">S.K. 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Liang</name></author><author><name sortKey="Wong, C H" uniqKey="Wong C">C.H. Wong</name></author></analytic></biblStruct></listBibl></div1></back></TEI><affiliations><list><country><li>Australie</li><li>République populaire de Chine</li></country></list><tree><country name="République populaire de Chine"><noRegion><name sortKey="Wang, Jian Min" sort="Wang, Jian Min" uniqKey="Wang J" first="Jian-Min" last="Wang">Jian-Min Wang</name></noRegion><name sortKey="Shi, Zheng Li" sort="Shi, Zheng Li" uniqKey="Shi Z" first="Zheng-Li" last="Shi">Zheng-Li Shi</name></country><country name="Australie"><noRegion><name sortKey="Wang, Lin Fa" sort="Wang, Lin Fa" uniqKey="Wang L" first="Lin-Fa" last="Wang">Lin-Fa Wang</name></noRegion></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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