Construction of a severe acute respiratory syndrome coronavirus infectious cDNA clone and a replicon to study coronavirus RNA synthesis
Identifieur interne : 004467 ( Main/Curation ); précédent : 004466; suivant : 004468Construction 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.
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)
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.
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Links to Exploration step
Pascal:06-0518585Le document en format XML
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<sourceDesc><biblStruct><analytic><title xml:lang="en" level="a">Construction of a severe acute respiratory syndrome coronavirus infectious cDNA clone and a replicon to study coronavirus RNA synthesis</title>
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</author>
<author><name sortKey="Alvarez, Enrique" sort="Alvarez, Enrique" uniqKey="Alvarez E" first="Enrique" last="Alvarez">Enrique Alvarez</name>
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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>
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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="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>
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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="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>
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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="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>
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<sZ>11 aut.</sZ>
<sZ>12 aut.</sZ>
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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="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>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="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>
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<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="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>
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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>
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<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>
<double idat="0022-538X:2006:Almazan F:construction:of:a"><INIST><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en" level="a">Construction of a severe acute respiratory syndrome coronavirus infectious cDNA clone and a replicon to study coronavirus RNA synthesis</title>
<author><name sortKey="Almazan, Fernando" sort="Almazan, Fernando" uniqKey="Almazan F" first="Fernando" last="Almazan">Fernando Almazan</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>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="Dediego, Marta L" sort="Dediego, Marta L" uniqKey="Dediego M" first="Marta L." last="Dediego">Marta L. Dediego</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>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="Galan, Carmen" sort="Galan, Carmen" uniqKey="Galan C" first="Carmen" last="Galan">Carmen Galan</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>10 aut.</sZ>
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<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="Escors, David" sort="Escors, David" uniqKey="Escors D" first="David" last="Escors">David Escors</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>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>
<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>
<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>
<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>
<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>
<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>
<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>
<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>
<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>
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<author><name sortKey="Enjuanes, Luis" sort="Enjuanes, Luis" uniqKey="Enjuanes L" first="Luis" last="Enjuanes">Luis Enjuanes</name>
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</titleStmt>
<publicationStmt><idno type="wicri:source">INIST</idno>
<idno type="inist">06-0518585</idno>
<date when="2006">2006</date>
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<sourceDesc><biblStruct><analytic><title xml:lang="en" level="a">Construction of a severe acute respiratory syndrome coronavirus infectious cDNA clone and a replicon to study coronavirus RNA synthesis</title>
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</placeName>
</affiliation>
</author>
<author><name sortKey="Dediego, Marta L" sort="Dediego, Marta L" uniqKey="Dediego M" first="Marta L." last="Dediego">Marta L. Dediego</name>
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<country>Espagne</country>
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</placeName>
</affiliation>
</author>
<author><name sortKey="Galan, Carmen" sort="Galan, Carmen" uniqKey="Galan C" first="Carmen" last="Galan">Carmen Galan</name>
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<country>Espagne</country>
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</affiliation>
</author>
<author><name sortKey="Escors, David" sort="Escors, David" uniqKey="Escors D" first="David" last="Escors">David Escors</name>
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<country>Espagne</country>
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</placeName>
</affiliation>
</author>
<author><name sortKey="Alvarez, Enrique" sort="Alvarez, Enrique" uniqKey="Alvarez E" first="Enrique" last="Alvarez">Enrique Alvarez</name>
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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>
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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="Sola, Isabel" sort="Sola, Isabel" uniqKey="Sola I" first="Isabel" last="Sola">Isabel Sola</name>
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<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="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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<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>
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<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="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>
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<s3>ESP</s3>
<sZ>1 aut.</sZ>
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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="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>
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<country>Espagne</country>
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<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>
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<country>Espagne</country>
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<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>
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<country>Espagne</country>
<placeName><settlement type="city">Madrid</settlement>
<region nuts="2" type="region">Communauté de Madrid</region>
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</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>Complementary DNA</term>
<term>Coronavirus</term>
<term>Microbiology</term>
<term>Severe acute respiratory syndrome</term>
<term>Virology</term>
</keywords>
<keywords scheme="Pascal" xml:lang="fr"><term>Coronavirus</term>
<term>DNA complémentaire</term>
<term>Microbiologie</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>
</INIST>
<PubMed><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Construction of a severe acute respiratory syndrome coronavirus infectious cDNA clone and a replicon to study coronavirus RNA synthesis.</title>
<author><name sortKey="Almazan, Fernando" sort="Almazan, Fernando" uniqKey="Almazan F" first="Fernando" last="Almazán">Fernando Almazán</name>
<affiliation wicri:level="2"><nlm:affiliation>Department of Molecular and Cell Biology, Centro Nacional de Biotecnología, CSIC, Darwin 3, Campus Universidad Autónoma, Cantoblanco, 28049 Madrid, Spain.</nlm:affiliation>
<country xml:lang="fr">Espagne</country>
<wicri:regionArea>Department of Molecular and Cell Biology, Centro Nacional de Biotecnología, CSIC, Darwin 3, Campus Universidad Autónoma, Cantoblanco, 28049 Madrid</wicri:regionArea>
<placeName><region nuts="2" type="communauté">Communauté de Madrid</region>
</placeName>
</affiliation>
</author>
<author><name sortKey="Dediego, Marta L" sort="Dediego, Marta L" uniqKey="Dediego M" first="Marta L" last="Dediego">Marta L. Dediego</name>
</author>
<author><name sortKey="Galan, Carmen" sort="Galan, Carmen" uniqKey="Galan C" first="Carmen" last="Galán">Carmen Galán</name>
</author>
<author><name sortKey="Escors, David" sort="Escors, David" uniqKey="Escors D" first="David" last="Escors">David Escors</name>
</author>
<author><name sortKey="Alvarez, Enrique" sort="Alvarez, Enrique" uniqKey="Alvarez E" first="Enrique" last="Alvarez">Enrique Alvarez</name>
</author>
<author><name sortKey="Ortego, Javier" sort="Ortego, Javier" uniqKey="Ortego J" first="Javier" last="Ortego">Javier Ortego</name>
</author>
<author><name sortKey="Sola, Isabel" sort="Sola, Isabel" uniqKey="Sola I" first="Isabel" last="Sola">Isabel Sola</name>
</author>
<author><name sortKey="Zu Iga, Sonia" sort="Zu Iga, Sonia" uniqKey="Zu Iga S" first="Sonia" last="Zu Iga">Sonia Zu Iga</name>
</author>
<author><name sortKey="Alonso, Sara" sort="Alonso, Sara" uniqKey="Alonso S" first="Sara" last="Alonso">Sara Alonso</name>
</author>
<author><name sortKey="Moreno, Jose L" sort="Moreno, Jose L" uniqKey="Moreno J" first="Jose L" last="Moreno">Jose L. Moreno</name>
</author>
<author><name sortKey="Nogales, Aitor" sort="Nogales, Aitor" uniqKey="Nogales A" first="Aitor" last="Nogales">Aitor Nogales</name>
</author>
<author><name sortKey="Capiscol, Carmen" sort="Capiscol, Carmen" uniqKey="Capiscol C" first="Carmen" last="Capiscol">Carmen Capiscol</name>
</author>
<author><name sortKey="Enjuanes, Luis" sort="Enjuanes, Luis" uniqKey="Enjuanes L" first="Luis" last="Enjuanes">Luis Enjuanes</name>
</author>
</titleStmt>
<publicationStmt><idno type="wicri:source">PubMed</idno>
<date when="2006">2006</date>
<idno type="RBID">pubmed:16928748</idno>
<idno type="pmid">16928748</idno>
<idno type="doi">10.1128/JVI.00385-06</idno>
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<sourceDesc><biblStruct><analytic><title xml:lang="en">Construction of a severe acute respiratory syndrome coronavirus infectious cDNA clone and a replicon to study coronavirus RNA synthesis.</title>
<author><name sortKey="Almazan, Fernando" sort="Almazan, Fernando" uniqKey="Almazan F" first="Fernando" last="Almazán">Fernando Almazán</name>
<affiliation wicri:level="2"><nlm:affiliation>Department of Molecular and Cell Biology, Centro Nacional de Biotecnología, CSIC, Darwin 3, Campus Universidad Autónoma, Cantoblanco, 28049 Madrid, Spain.</nlm:affiliation>
<country xml:lang="fr">Espagne</country>
<wicri:regionArea>Department of Molecular and Cell Biology, Centro Nacional de Biotecnología, CSIC, Darwin 3, Campus Universidad Autónoma, Cantoblanco, 28049 Madrid</wicri:regionArea>
<placeName><region nuts="2" type="communauté">Communauté de Madrid</region>
</placeName>
</affiliation>
</author>
<author><name sortKey="Dediego, Marta L" sort="Dediego, Marta L" uniqKey="Dediego M" first="Marta L" last="Dediego">Marta L. Dediego</name>
</author>
<author><name sortKey="Galan, Carmen" sort="Galan, Carmen" uniqKey="Galan C" first="Carmen" last="Galán">Carmen Galán</name>
</author>
<author><name sortKey="Escors, David" sort="Escors, David" uniqKey="Escors D" first="David" last="Escors">David Escors</name>
</author>
<author><name sortKey="Alvarez, Enrique" sort="Alvarez, Enrique" uniqKey="Alvarez E" first="Enrique" last="Alvarez">Enrique Alvarez</name>
</author>
<author><name sortKey="Ortego, Javier" sort="Ortego, Javier" uniqKey="Ortego J" first="Javier" last="Ortego">Javier Ortego</name>
</author>
<author><name sortKey="Sola, Isabel" sort="Sola, Isabel" uniqKey="Sola I" first="Isabel" last="Sola">Isabel Sola</name>
</author>
<author><name sortKey="Zu Iga, Sonia" sort="Zu Iga, Sonia" uniqKey="Zu Iga S" first="Sonia" last="Zu Iga">Sonia Zu Iga</name>
</author>
<author><name sortKey="Alonso, Sara" sort="Alonso, Sara" uniqKey="Alonso S" first="Sara" last="Alonso">Sara Alonso</name>
</author>
<author><name sortKey="Moreno, Jose L" sort="Moreno, Jose L" uniqKey="Moreno J" first="Jose L" last="Moreno">Jose L. Moreno</name>
</author>
<author><name sortKey="Nogales, Aitor" sort="Nogales, Aitor" uniqKey="Nogales A" first="Aitor" last="Nogales">Aitor Nogales</name>
</author>
<author><name sortKey="Capiscol, Carmen" sort="Capiscol, Carmen" uniqKey="Capiscol C" first="Carmen" last="Capiscol">Carmen Capiscol</name>
</author>
<author><name sortKey="Enjuanes, Luis" sort="Enjuanes, Luis" uniqKey="Enjuanes L" first="Luis" last="Enjuanes">Luis Enjuanes</name>
</author>
</analytic>
<series><title level="j">Journal of virology</title>
<idno type="ISSN">0022-538X</idno>
<imprint><date when="2006" type="published">2006</date>
</imprint>
</series>
</biblStruct>
</sourceDesc>
</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>Cricetinae</term>
<term>DNA, Complementary (genetics)</term>
<term>DNA, Viral (genetics)</term>
<term>Humans</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>Vero Cells</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="MESH" xml:lang="fr"><term>Animaux</term>
<term>Cellules Vero</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>
</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 methyltransferase 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>
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