SrasV1, Main, Exploration, bibRecord, 004467

Construction of a severe acute respiratory syndrome coronavirus infectious cDNA clone and a replicon to study coronavirus RNA synthesis

Identifieur interne : 004467 ( Main/Exploration ); précédent : 004466; suivant : 004468

Construction of a severe acute respiratory syndrome coronavirus infectious cDNA clone and a replicon to study coronavirus RNA synthesis

Auteurs : Fernando Almazan [Espagne] ; Marta L. Dediego [Espagne] ; Carmen Galan [Espagne] ; David Escors [Espagne] ; Enrique Alvarez [Espagne] ; Javier Ortego [Espagne] ; Isabel Sola [Espagne] ; Sonia Zumiga [Espagne] ; Sara Alonso [Espagne] ; Jose L. Moreno [Espagne] ; Aitor Nogales [Espagne] ; Carmen Capiscol [Espagne] ; Luis Enjuanes [Espagne]

Source :

Journal of virology [ 0022-538X ] ; 2006.

RBID : Pascal:06-0518585

Descripteurs français

KwdFr :
- ADN complémentaire (génétique), ADN viral (génétique), ARN viral (biosynthèse), ARN viral (génétique), Animaux, Cellules Vero, Chromosomes artificiels de bactérie (génétique), Clonage moléculaire, Cricetinae, Données de séquences moléculaires, Humains, Lignée cellulaire, Réplicon, Séquence nucléotidique, Virus du SRAS (génétique), Virus du SRAS (métabolisme).
MESH :
- biosynthèse : ARN viral.
- génétique : ADN complémentaire, ADN viral, ARN viral, Chromosomes artificiels de bactérie, Virus du SRAS.
- métabolisme : Virus du SRAS.
Pascal (Inist)
- Animaux, Cellules Vero, Clonage moléculaire, Coronavirus, Cricetinae, DNA complémentaire, Données de séquences moléculaires, Humains, Lignée cellulaire, Microbiologie, Réplicon, Séquence nucléotidique, Virologie, Syndrome respiratoire aigu sévère.

English descriptors

KwdEn :
- Animals, Base Sequence, Cell Line, Chlorocebus aethiops, Chromosomes, Artificial, Bacterial (genetics), Cloning, Molecular, Complementary DNA, Coronavirus, Cricetinae, DNA, Complementary (genetics), DNA, Viral (genetics), Humans, Microbiology, Molecular Sequence Data, RNA, Viral (biosynthesis), RNA, Viral (genetics), Replicon, SARS Virus (genetics), SARS Virus (metabolism), Severe acute respiratory syndrome, Vero Cells, Virology.
MESH :
- chemical , biosynthesis : RNA, Viral.
- chemical , genetics : DNA, Complementary, DNA, Viral, RNA, Viral.
- genetics : Chromosomes, Artificial, Bacterial, SARS Virus.
- metabolism : SARS Virus.
- Animals, Base Sequence, Cell Line, Chlorocebus aethiops, Cloning, Molecular, Cricetinae, Humans, Molecular Sequence Data, Replicon, Vero Cells.

Abstract

The engineering of a full-length infectious cDNA clone and a functional replicon of the severe acute respiratory syndrome coronavirus (SARS-CoV) Urbani strain as bacterial artificial chromosomes (BACs) is described in this study. In this system, the viral RNA was expressed in the cell nucleus under the control of the cytomegalovirus promoter and further amplified in the cytoplasm by the viral replicase. Both the infectious clone and the replicon were fully stable in Escherichia coli. Using the SARS-CoV replicon, we have shown that the recently described RNA-processing enzymes exoribonuclease, endoribonuclease, and 2'-O-ribose methyl-transferase were essential for efficient coronavirus RNA synthesis. The SARS reverse genetic system developed as a BAC constitutes a useful tool for the study of fundamental viral processes and also for developing genetically defined vaccines.

Affiliations:

Le document en format XML

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<author><name sortKey="Alvarez, Enrique" sort="Alvarez, Enrique" uniqKey="Alvarez E" first="Enrique" last="Alvarez">Enrique Alvarez</name>
<affiliation wicri:level="3"><inist:fA14 i1="01"><s1>Centro Nacional de Biotecnología, CSIC, Department of Molecular and Cell Biology, Darwin 3, Campus Universidad Autónoma, Cantoblanco</s1>
<s2>Madrid</s2>
<s3>ESP</s3>
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<country>Espagne</country>
<placeName><settlement type="city">Madrid</settlement>
<region nuts="2" type="region">Communauté de Madrid</region>
</placeName>
</affiliation>
</author>
<author><name sortKey="Ortego, Javier" sort="Ortego, Javier" uniqKey="Ortego J" first="Javier" last="Ortego">Javier Ortego</name>
<affiliation wicri:level="3"><inist:fA14 i1="01"><s1>Centro Nacional de Biotecnología, CSIC, Department of Molecular and Cell Biology, Darwin 3, Campus Universidad Autónoma, Cantoblanco</s1>
<s2>Madrid</s2>
<s3>ESP</s3>
<sZ>1 aut.</sZ>
<sZ>2 aut.</sZ>
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<sZ>4 aut.</sZ>
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<sZ>11 aut.</sZ>
<sZ>12 aut.</sZ>
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</inist:fA14>
<country>Espagne</country>
<placeName><settlement type="city">Madrid</settlement>
<region nuts="2" type="region">Communauté de Madrid</region>
</placeName>
</affiliation>
</author>
<author><name sortKey="Sola, Isabel" sort="Sola, Isabel" uniqKey="Sola I" first="Isabel" last="Sola">Isabel Sola</name>
<affiliation wicri:level="3"><inist:fA14 i1="01"><s1>Centro Nacional de Biotecnología, CSIC, Department of Molecular and Cell Biology, Darwin 3, Campus Universidad Autónoma, Cantoblanco</s1>
<s2>Madrid</s2>
<s3>ESP</s3>
<sZ>1 aut.</sZ>
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</inist:fA14>
<country>Espagne</country>
<placeName><settlement type="city">Madrid</settlement>
<region nuts="2" type="region">Communauté de Madrid</region>
</placeName>
</affiliation>
</author>
<author><name sortKey="Zumiga, Sonia" sort="Zumiga, Sonia" uniqKey="Zumiga S" first="Sonia" last="Zumiga">Sonia Zumiga</name>
<affiliation wicri:level="3"><inist:fA14 i1="01"><s1>Centro Nacional de Biotecnología, CSIC, Department of Molecular and Cell Biology, Darwin 3, Campus Universidad Autónoma, Cantoblanco</s1>
<s2>Madrid</s2>
<s3>ESP</s3>
<sZ>1 aut.</sZ>
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</inist:fA14>
<country>Espagne</country>
<placeName><settlement type="city">Madrid</settlement>
<region nuts="2" type="region">Communauté de Madrid</region>
</placeName>
</affiliation>
</author>
<author><name sortKey="Alonso, Sara" sort="Alonso, Sara" uniqKey="Alonso S" first="Sara" last="Alonso">Sara Alonso</name>
<affiliation wicri:level="3"><inist:fA14 i1="01"><s1>Centro Nacional de Biotecnología, CSIC, Department of Molecular and Cell Biology, Darwin 3, Campus Universidad Autónoma, Cantoblanco</s1>
<s2>Madrid</s2>
<s3>ESP</s3>
<sZ>1 aut.</sZ>
<sZ>2 aut.</sZ>
<sZ>3 aut.</sZ>
<sZ>4 aut.</sZ>
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<sZ>12 aut.</sZ>
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</inist:fA14>
<country>Espagne</country>
<placeName><settlement type="city">Madrid</settlement>
<region nuts="2" type="region">Communauté de Madrid</region>
</placeName>
</affiliation>
</author>
<author><name sortKey="Moreno, Jose L" sort="Moreno, Jose L" uniqKey="Moreno J" first="Jose L." last="Moreno">Jose L. Moreno</name>
<affiliation wicri:level="3"><inist:fA14 i1="01"><s1>Centro Nacional de Biotecnología, CSIC, Department of Molecular and Cell Biology, Darwin 3, Campus Universidad Autónoma, Cantoblanco</s1>
<s2>Madrid</s2>
<s3>ESP</s3>
<sZ>1 aut.</sZ>
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<sZ>4 aut.</sZ>
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<sZ>11 aut.</sZ>
<sZ>12 aut.</sZ>
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</inist:fA14>
<country>Espagne</country>
<placeName><settlement type="city">Madrid</settlement>
<region nuts="2" type="region">Communauté de Madrid</region>
</placeName>
</affiliation>
</author>
<author><name sortKey="Nogales, Aitor" sort="Nogales, Aitor" uniqKey="Nogales A" first="Aitor" last="Nogales">Aitor Nogales</name>
<affiliation wicri:level="3"><inist:fA14 i1="01"><s1>Centro Nacional de Biotecnología, CSIC, Department of Molecular and Cell Biology, Darwin 3, Campus Universidad Autónoma, Cantoblanco</s1>
<s2>Madrid</s2>
<s3>ESP</s3>
<sZ>1 aut.</sZ>
<sZ>2 aut.</sZ>
<sZ>3 aut.</sZ>
<sZ>4 aut.</sZ>
<sZ>5 aut.</sZ>
<sZ>6 aut.</sZ>
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<sZ>8 aut.</sZ>
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<sZ>10 aut.</sZ>
<sZ>11 aut.</sZ>
<sZ>12 aut.</sZ>
<sZ>13 aut.</sZ>
</inist:fA14>
<country>Espagne</country>
<placeName><settlement type="city">Madrid</settlement>
<region nuts="2" type="region">Communauté de Madrid</region>
</placeName>
</affiliation>
</author>
<author><name sortKey="Capiscol, Carmen" sort="Capiscol, Carmen" uniqKey="Capiscol C" first="Carmen" last="Capiscol">Carmen Capiscol</name>
<affiliation wicri:level="3"><inist:fA14 i1="01"><s1>Centro Nacional de Biotecnología, CSIC, Department of Molecular and Cell Biology, Darwin 3, Campus Universidad Autónoma, Cantoblanco</s1>
<s2>Madrid</s2>
<s3>ESP</s3>
<sZ>1 aut.</sZ>
<sZ>2 aut.</sZ>
<sZ>3 aut.</sZ>
<sZ>4 aut.</sZ>
<sZ>5 aut.</sZ>
<sZ>6 aut.</sZ>
<sZ>7 aut.</sZ>
<sZ>8 aut.</sZ>
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<sZ>10 aut.</sZ>
<sZ>11 aut.</sZ>
<sZ>12 aut.</sZ>
<sZ>13 aut.</sZ>
</inist:fA14>
<country>Espagne</country>
<placeName><settlement type="city">Madrid</settlement>
<region nuts="2" type="region">Communauté de Madrid</region>
</placeName>
</affiliation>
</author>
<author><name sortKey="Enjuanes, Luis" sort="Enjuanes, Luis" uniqKey="Enjuanes L" first="Luis" last="Enjuanes">Luis Enjuanes</name>
<affiliation wicri:level="3"><inist:fA14 i1="01"><s1>Centro Nacional de Biotecnología, CSIC, Department of Molecular and Cell Biology, Darwin 3, Campus Universidad Autónoma, Cantoblanco</s1>
<s2>Madrid</s2>
<s3>ESP</s3>
<sZ>1 aut.</sZ>
<sZ>2 aut.</sZ>
<sZ>3 aut.</sZ>
<sZ>4 aut.</sZ>
<sZ>5 aut.</sZ>
<sZ>6 aut.</sZ>
<sZ>7 aut.</sZ>
<sZ>8 aut.</sZ>
<sZ>9 aut.</sZ>
<sZ>10 aut.</sZ>
<sZ>11 aut.</sZ>
<sZ>12 aut.</sZ>
<sZ>13 aut.</sZ>
</inist:fA14>
<country>Espagne</country>
<placeName><settlement type="city">Madrid</settlement>
<region nuts="2" type="region">Communauté de Madrid</region>
</placeName>
</affiliation>
</author>
</analytic>
<series><title level="j" type="main">Journal of virology</title>
<title level="j" type="abbreviated">J. virol.</title>
<idno type="ISSN">0022-538X</idno>
<imprint><date when="2006">2006</date>
</imprint>
</series>
</biblStruct>
</sourceDesc>
<seriesStmt><title level="j" type="main">Journal of virology</title>
<title level="j" type="abbreviated">J. virol.</title>
<idno type="ISSN">0022-538X</idno>
</seriesStmt>
</fileDesc>
<profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Animals</term>
<term>Base Sequence</term>
<term>Cell Line</term>
<term>Chlorocebus aethiops</term>
<term>Chromosomes, Artificial, Bacterial (genetics)</term>
<term>Cloning, Molecular</term>
<term>Complementary DNA</term>
<term>Coronavirus</term>
<term>Cricetinae</term>
<term>DNA, Complementary (genetics)</term>
<term>DNA, Viral (genetics)</term>
<term>Humans</term>
<term>Microbiology</term>
<term>Molecular Sequence Data</term>
<term>RNA, Viral (biosynthesis)</term>
<term>RNA, Viral (genetics)</term>
<term>Replicon</term>
<term>SARS Virus (genetics)</term>
<term>SARS Virus (metabolism)</term>
<term>Severe acute respiratory syndrome</term>
<term>Vero Cells</term>
<term>Virology</term>
</keywords>
<keywords scheme="KwdFr" xml:lang="fr"><term>ADN complémentaire (génétique)</term>
<term>ADN viral (génétique)</term>
<term>ARN viral (biosynthèse)</term>
<term>ARN viral (génétique)</term>
<term>Animaux</term>
<term>Cellules Vero</term>
<term>Chromosomes artificiels de bactérie (génétique)</term>
<term>Clonage moléculaire</term>
<term>Cricetinae</term>
<term>Données de séquences moléculaires</term>
<term>Humains</term>
<term>Lignée cellulaire</term>
<term>Réplicon</term>
<term>Séquence nucléotidique</term>
<term>Virus du SRAS (génétique)</term>
<term>Virus du SRAS (métabolisme)</term>
</keywords>
<keywords scheme="MESH" type="chemical" qualifier="biosynthesis" xml:lang="en"><term>RNA, Viral</term>
</keywords>
<keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>DNA, Complementary</term>
<term>DNA, Viral</term>
<term>RNA, Viral</term>
</keywords>
<keywords scheme="MESH" qualifier="biosynthèse" xml:lang="fr"><term>ARN viral</term>
</keywords>
<keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Chromosomes, Artificial, Bacterial</term>
<term>SARS Virus</term>
</keywords>
<keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>ADN complémentaire</term>
<term>ADN viral</term>
<term>ARN viral</term>
<term>Chromosomes artificiels de bactérie</term>
<term>Virus du SRAS</term>
</keywords>
<keywords scheme="MESH" qualifier="metabolism" xml:lang="en"><term>SARS Virus</term>
</keywords>
<keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>Virus du SRAS</term>
</keywords>
<keywords scheme="MESH" xml:lang="en"><term>Animals</term>
<term>Base Sequence</term>
<term>Cell Line</term>
<term>Chlorocebus aethiops</term>
<term>Cloning, Molecular</term>
<term>Cricetinae</term>
<term>Humans</term>
<term>Molecular Sequence Data</term>
<term>Replicon</term>
<term>Vero Cells</term>
</keywords>
<keywords scheme="Pascal" xml:lang="fr"><term>Animaux</term>
<term>Cellules Vero</term>
<term>Clonage moléculaire</term>
<term>Coronavirus</term>
<term>Cricetinae</term>
<term>DNA complémentaire</term>
<term>Données de séquences moléculaires</term>
<term>Humains</term>
<term>Lignée cellulaire</term>
<term>Microbiologie</term>
<term>Réplicon</term>
<term>Séquence nucléotidique</term>
<term>Virologie</term>
<term>Syndrome respiratoire aigu sévère</term>
</keywords>
</textClass>
</profileDesc>
</teiHeader>
<front><div type="abstract" xml:lang="en">The engineering of a full-length infectious cDNA clone and a functional replicon of the severe acute respiratory syndrome coronavirus (SARS-CoV) Urbani strain as bacterial artificial chromosomes (BACs) is described in this study. In this system, the viral RNA was expressed in the cell nucleus under the control of the cytomegalovirus promoter and further amplified in the cytoplasm by the viral replicase. Both the infectious clone and the replicon were fully stable in Escherichia coli. Using the SARS-CoV replicon, we have shown that the recently described RNA-processing enzymes exoribonuclease, endoribonuclease, and 2'-O-ribose methyl-transferase were essential for efficient coronavirus RNA synthesis. The SARS reverse genetic system developed as a BAC constitutes a useful tool for the study of fundamental viral processes and also for developing genetically defined vaccines.</div>
</front>
</TEI>
<affiliations><list><country><li>Espagne</li>
</country>
<region><li>Communauté de Madrid</li>
</region>
<settlement><li>Madrid</li>
</settlement>
</list>
<tree><country name="Espagne"><region name="Communauté de Madrid"><name sortKey="Almazan, Fernando" sort="Almazan, Fernando" uniqKey="Almazan F" first="Fernando" last="Almazan">Fernando Almazan</name>
</region>
<name sortKey="Alonso, Sara" sort="Alonso, Sara" uniqKey="Alonso S" first="Sara" last="Alonso">Sara Alonso</name>
<name sortKey="Alvarez, Enrique" sort="Alvarez, Enrique" uniqKey="Alvarez E" first="Enrique" last="Alvarez">Enrique Alvarez</name>
<name sortKey="Capiscol, Carmen" sort="Capiscol, Carmen" uniqKey="Capiscol C" first="Carmen" last="Capiscol">Carmen Capiscol</name>
<name sortKey="Dediego, Marta L" sort="Dediego, Marta L" uniqKey="Dediego M" first="Marta L." last="Dediego">Marta L. Dediego</name>
<name sortKey="Enjuanes, Luis" sort="Enjuanes, Luis" uniqKey="Enjuanes L" first="Luis" last="Enjuanes">Luis Enjuanes</name>
<name sortKey="Escors, David" sort="Escors, David" uniqKey="Escors D" first="David" last="Escors">David Escors</name>
<name sortKey="Galan, Carmen" sort="Galan, Carmen" uniqKey="Galan C" first="Carmen" last="Galan">Carmen Galan</name>
<name sortKey="Moreno, Jose L" sort="Moreno, Jose L" uniqKey="Moreno J" first="Jose L." last="Moreno">Jose L. Moreno</name>
<name sortKey="Nogales, Aitor" sort="Nogales, Aitor" uniqKey="Nogales A" first="Aitor" last="Nogales">Aitor Nogales</name>
<name sortKey="Ortego, Javier" sort="Ortego, Javier" uniqKey="Ortego J" first="Javier" last="Ortego">Javier Ortego</name>
<name sortKey="Sola, Isabel" sort="Sola, Isabel" uniqKey="Sola I" first="Isabel" last="Sola">Isabel Sola</name>
<name sortKey="Zumiga, Sonia" sort="Zumiga, Sonia" uniqKey="Zumiga S" first="Sonia" last="Zumiga">Sonia Zumiga</name>
</country>
</tree>
</affiliations>
</record>

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Construction of a severe acute respiratory syndrome coronavirus infectious cDNA clone and a replicon to study coronavirus RNA synthesis

Construction of a severe acute respiratory syndrome coronavirus infectious cDNA clone and a replicon to study coronavirus RNA synthesis

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