Generation of Full-Length Infectious cDNA Clones of Middle East Respiratory Syndrome Coronavirus.
Identifieur interne : 000561 ( Main/Exploration ); précédent : 000560; suivant : 000562Generation of Full-Length Infectious cDNA Clones of Middle East Respiratory Syndrome Coronavirus.
Auteurs : Jeong Lee [Corée du Sud] ; Sojung Bae [Corée du Sud] ; Jinjong Myoung [Corée du Sud]Source :
- Journal of microbiology and biotechnology [ 1738-8872 ] ; 2019.
Descripteurs français
- KwdFr :
- ADN complémentaire (génétique), ARN viral (génétique), Coronavirus du syndrome respiratoire du Moyen-Orient (génétique), Coronavirus du syndrome respiratoire du Moyen-Orient (isolement et purification), Gènes viraux (génétique), Génie génétique, Génome viral (génétique), Génétique inverse, Humains, Infections à coronavirus (virologie), Transcription génétique.
- MESH :
- génétique : ADN complémentaire, ARN viral, Coronavirus du syndrome respiratoire du Moyen-Orient, Gènes viraux, Génome viral.
- isolement et purification : Coronavirus du syndrome respiratoire du Moyen-Orient.
- virologie : Infections à coronavirus.
- Génie génétique, Génétique inverse, Humains, Transcription génétique.
English descriptors
- KwdEn :
- Coronavirus Infections (virology), DNA, Complementary (genetics), Genes, Viral (genetics), Genetic Engineering, Genome, Viral (genetics), Humans, Middle East Respiratory Syndrome Coronavirus (genetics), Middle East Respiratory Syndrome Coronavirus (isolation & purification), RNA, Viral (genetics), Reverse Genetics, Transcription, Genetic.
- MESH :
- chemical , genetics : DNA, Complementary, RNA, Viral.
- genetics : Genes, Viral, Genome, Viral, Middle East Respiratory Syndrome Coronavirus.
- isolation & purification : Middle East Respiratory Syndrome Coronavirus.
- virology : Coronavirus Infections.
- Genetic Engineering, Humans, Reverse Genetics, Transcription, Genetic.
Abstract
Middle East respiratory syndrome coronavirus (MERS-CoV) was first identified in Saudi Arabia in 2012 and related infection cases have been reported in over 20 countries. Roughly 10,000 human cases have so far been reported in total with fatality rates at up to 40%. The majority of cases have occurred in Saudi Arabia with mostly sporadic outbreaks outside the country except for the one in South Korea in 2015. The Korean MERS-CoV strain was isolated from the second Korean patient and its genome was fully sequenced and deposited. To develop virusspecific protective and therapeutic agents against the Korean isolate and to investigate molecular determinants of virus-host interactions, it is of paramount importance to generate its full-length cDNA. Here we report that two full-length cDNAs from a Korean patientisolated MERS-CoV strain were generated by a combination of conventional cloning techniques and efficient Gibson assembly reactions. The full-length cDNAs were validated by restriction analysis and their sequence was verified by Sanger method. The resulting cDNA was efficiently transcribed in vitro and the T7 promoter-driven expression was robust. The resulting reverse genetic system will add to the published list of MERS-CoV cDNAs and facilitate the development of Korean isolate-specific antiviral measures.
DOI: 10.4014/jmb.0905.05061
PubMed: 31154749
Affiliations:
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Le document en format XML
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of Korea.</nlm:affiliation><country xml:lang="fr">Corée du Sud</country><wicri:regionArea>Korea Zoonosis Research Institute and Genetic Engineering Research Institute, Chonbuk National University, Jeonju 561-756</wicri:regionArea><wicri:noRegion>Jeonju 561-756</wicri:noRegion></affiliation></author><author><name sortKey="Myoung, Jinjong" sort="Myoung, Jinjong" uniqKey="Myoung J" first="Jinjong" last="Myoung">Jinjong Myoung</name><affiliation wicri:level="1"><nlm:affiliation>Korea Zoonosis Research Institute and Genetic Engineering Research Institute, Chonbuk National University, Jeonju 561-756, Republic of Korea.</nlm:affiliation><country xml:lang="fr">Corée du Sud</country><wicri:regionArea>Korea Zoonosis Research Institute and Genetic Engineering Research Institute, Chonbuk National University, Jeonju 561-756</wicri:regionArea><wicri:noRegion>Jeonju 561-756</wicri:noRegion></affiliation></author></analytic><series><title level="j">Journal of microbiology and 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viraux (génétique)</term><term>Génie génétique</term><term>Génome viral (génétique)</term><term>Génétique inverse</term><term>Humains</term><term>Infections à coronavirus (virologie)</term><term>Transcription génétique</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>DNA, Complementary</term><term>RNA, Viral</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Genes, Viral</term><term>Genome, Viral</term><term>Middle East Respiratory Syndrome Coronavirus</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>ADN complémentaire</term><term>ARN viral</term><term>Coronavirus du syndrome respiratoire du Moyen-Orient</term><term>Gènes viraux</term><term>Génome viral</term></keywords><keywords scheme="MESH" qualifier="isolation & purification" xml:lang="en"><term>Middle East Respiratory Syndrome Coronavirus</term></keywords><keywords scheme="MESH" qualifier="isolement et purification" xml:lang="fr"><term>Coronavirus du syndrome respiratoire du Moyen-Orient</term></keywords><keywords scheme="MESH" qualifier="virologie" xml:lang="fr"><term>Infections à coronavirus</term></keywords><keywords scheme="MESH" qualifier="virology" xml:lang="en"><term>Coronavirus Infections</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Genetic Engineering</term><term>Humans</term><term>Reverse Genetics</term><term>Transcription, Genetic</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Génie génétique</term><term>Génétique inverse</term><term>Humains</term><term>Transcription génétique</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Middle East respiratory syndrome coronavirus (MERS-CoV) was first identified in Saudi Arabia in 2012 and related infection cases have been reported in over 20 countries. Roughly 10,000 human cases have so far been reported in total with fatality rates at up to 40%. The majority of cases have occurred in Saudi Arabia with mostly sporadic outbreaks outside the country except for the one in South Korea in 2015. The Korean MERS-CoV strain was isolated from the second Korean patient and its genome was fully sequenced and deposited. To develop virusspecific protective and therapeutic agents against the Korean isolate and to investigate molecular determinants of virus-host interactions, it is of paramount importance to generate its full-length cDNA. Here we report that two full-length cDNAs from a Korean patientisolated MERS-CoV strain were generated by a combination of conventional cloning techniques and efficient Gibson assembly reactions. The full-length cDNAs were validated by restriction analysis and their sequence was verified by Sanger method. The resulting cDNA was efficiently transcribed in vitro and the T7 promoter-driven expression was robust. The resulting reverse genetic system will add to the published list of MERS-CoV cDNAs and facilitate the development of Korean isolate-specific antiviral measures.</div></front></TEI><affiliations><list><country><li>Corée du Sud</li></country></list><tree><country name="Corée du Sud"><noRegion><name sortKey="Lee, Jeong" sort="Lee, Jeong" uniqKey="Lee J" first="Jeong" last="Lee">Jeong Lee</name></noRegion><name sortKey="Bae, Sojung" sort="Bae, Sojung" uniqKey="Bae S" first="Sojung" last="Bae">Sojung Bae</name><name sortKey="Myoung, Jinjong" sort="Myoung, Jinjong" uniqKey="Myoung J" first="Jinjong" last="Myoung">Jinjong Myoung</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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