Rapid one-step construction of a Middle East Respiratory Syndrome (MERS-CoV) infectious clone system by homologous recombination
Identifieur interne : 001722 ( Ncbi/Curation ); précédent : 001721; suivant : 001723Rapid one-step construction of a Middle East Respiratory Syndrome (MERS-CoV) infectious clone system by homologous recombination
Auteurs : Aidan M. Nikiforuk [Canada] ; Anders Leung [Canada] ; Bradley W. M. Cook [Canada] ; Deborah A. Court [Canada] ; Darwyn Kobasa [Canada] ; Steven S. Theriault [Canada]Source :
- Journal of Virological Methods [ 0166-0934 ] ; 2016.
Descripteurs français
- KwdFr :
- MESH :
English descriptors
- KwdEn :
- MESH :
- genetics : Middle East Respiratory Syndrome Coronavirus.
- methods : Genetic Engineering, Reverse Genetics.
- physiology : Middle East Respiratory Syndrome Coronavirus.
- Animals, Cell Line, Homologous Recombination.
Abstract
This work describes a minimalist approach to the construction and validation of a coronavirus reverse genetics system utilizing homologous recombination for vector construction and a variety of molecular techniques for virus detection. Homologous recombination in Detection of rescued virus by molecular assay (i.e.: RT-PCR) is a strong alternative to the more traditional immunological detection assays. Rescued MERS-CoV was genomically and phenotypically similar to the original isolate MERS-CoV/EMC-2012.
Url:
DOI: 10.1016/j.jviromet.2016.07.022
PubMed: 27459876
PubMed Central: 7113859
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PMC:7113859Le document en format XML
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Nikiforuk</name><affiliation wicri:level="1"><nlm:aff id="aff0005">Applied Biosafety Research Program, National Microbiology Laboratory at the Canadian Science Centre for Human and Animal Health, Public Health Agency of Canada, 1015 Arlington Street, Winnipeg, Manitoba, R3E 3P6, Canada</nlm:aff><country xml:lang="fr">Canada</country><wicri:regionArea>Applied Biosafety Research Program, National Microbiology Laboratory at the Canadian Science Centre for Human and Animal Health, Public Health Agency of Canada, 1015 Arlington Street, Winnipeg, Manitoba, R3E 3P6</wicri:regionArea><wicri:noRegion>R3E 3P6</wicri:noRegion></affiliation><affiliation wicri:level="1"><nlm:aff id="aff0010">National Microbiology Laboratory at the J. C. Wilt Infectious Diseases Research Centre, Public Health Agency of Canada, 745 Logan Street, Winnipeg, Manitoba, R3E 3L5, Canada</nlm:aff><country xml:lang="fr">Canada</country><wicri:regionArea>National Microbiology Laboratory at the J. C. Wilt Infectious Diseases Research Centre, Public Health Agency of Canada, 745 Logan Street, Winnipeg, Manitoba, R3E 3L5</wicri:regionArea><wicri:noRegion>R3E 3L5</wicri:noRegion></affiliation><affiliation wicri:level="1"><nlm:aff id="aff0015">High Containment Respiratory Viruses Group, Special Pathogens Program, National Microbiology Laboratory at the Canadian Science Centre for Human and Animal Health, Public Health Agency of Canada, 1015 Arlington Street, Winnipeg, Manitoba, R3E 3R2, Canada</nlm:aff><country xml:lang="fr">Canada</country><wicri:regionArea>High Containment Respiratory Viruses Group, Special Pathogens Program, National Microbiology Laboratory at the Canadian Science Centre for Human and Animal Health, Public Health Agency of Canada, 1015 Arlington Street, Winnipeg, Manitoba, R3E 3R2</wicri:regionArea><wicri:noRegion>R3E 3R2</wicri:noRegion></affiliation><affiliation wicri:level="4"><nlm:aff id="aff0020">Department of Microbiology, The University of Manitoba, Winnipeg, Manitoba, R3T 2N2, Canada</nlm:aff><country xml:lang="fr">Canada</country><wicri:regionArea>Department of Microbiology, The University of Manitoba, Winnipeg, Manitoba, R3T 2N2</wicri:regionArea><orgName type="university">Université du Manitoba</orgName><placeName><settlement type="city">Winnipeg</settlement><region type="state">Manitoba</region></placeName></affiliation></author><author><name sortKey="Leung, Anders" sort="Leung, Anders" uniqKey="Leung A" first="Anders" last="Leung">Anders Leung</name><affiliation wicri:level="1"><nlm:aff id="aff0015">High Containment Respiratory Viruses Group, Special Pathogens Program, National Microbiology Laboratory at the Canadian Science Centre for Human and Animal Health, Public Health Agency of Canada, 1015 Arlington Street, Winnipeg, Manitoba, R3E 3R2, Canada</nlm:aff><country xml:lang="fr">Canada</country><wicri:regionArea>High Containment Respiratory Viruses Group, Special Pathogens Program, National Microbiology Laboratory at the Canadian Science Centre for Human and Animal Health, Public Health Agency of Canada, 1015 Arlington Street, Winnipeg, Manitoba, R3E 3R2</wicri:regionArea><wicri:noRegion>R3E 3R2</wicri:noRegion></affiliation></author><author><name sortKey="Cook, Bradley W M" sort="Cook, Bradley W M" uniqKey="Cook B" first="Bradley W. M." last="Cook">Bradley W. M. Cook</name><affiliation wicri:level="1"><nlm:aff id="aff0005">Applied Biosafety Research Program, National Microbiology Laboratory at the Canadian Science Centre for Human and Animal Health, Public Health Agency of Canada, 1015 Arlington Street, Winnipeg, Manitoba, R3E 3P6, Canada</nlm:aff><country xml:lang="fr">Canada</country><wicri:regionArea>Applied Biosafety Research Program, National Microbiology Laboratory at the Canadian Science Centre for Human and Animal Health, Public Health Agency of Canada, 1015 Arlington Street, Winnipeg, Manitoba, R3E 3P6</wicri:regionArea><wicri:noRegion>R3E 3P6</wicri:noRegion></affiliation><affiliation wicri:level="1"><nlm:aff id="aff0010">National Microbiology Laboratory at the J. C. Wilt Infectious Diseases Research Centre, Public Health Agency of Canada, 745 Logan Street, Winnipeg, Manitoba, R3E 3L5, Canada</nlm:aff><country xml:lang="fr">Canada</country><wicri:regionArea>National Microbiology Laboratory at the J. C. Wilt Infectious Diseases Research Centre, Public Health Agency of Canada, 745 Logan Street, Winnipeg, Manitoba, R3E 3L5</wicri:regionArea><wicri:noRegion>R3E 3L5</wicri:noRegion></affiliation><affiliation wicri:level="4"><nlm:aff id="aff0020">Department of Microbiology, The University of Manitoba, Winnipeg, Manitoba, R3T 2N2, Canada</nlm:aff><country xml:lang="fr">Canada</country><wicri:regionArea>Department of Microbiology, The University of Manitoba, Winnipeg, Manitoba, R3T 2N2</wicri:regionArea><orgName type="university">Université du Manitoba</orgName><placeName><settlement type="city">Winnipeg</settlement><region type="state">Manitoba</region></placeName></affiliation></author><author><name sortKey="Court, Deborah A" sort="Court, Deborah A" uniqKey="Court D" first="Deborah A." last="Court">Deborah A. Court</name><affiliation wicri:level="4"><nlm:aff id="aff0020">Department of Microbiology, The University of Manitoba, Winnipeg, Manitoba, R3T 2N2, Canada</nlm:aff><country xml:lang="fr">Canada</country><wicri:regionArea>Department of Microbiology, The University of Manitoba, Winnipeg, Manitoba, R3T 2N2</wicri:regionArea><orgName type="university">Université du Manitoba</orgName><placeName><settlement type="city">Winnipeg</settlement><region type="state">Manitoba</region></placeName></affiliation></author><author><name sortKey="Kobasa, Darwyn" sort="Kobasa, Darwyn" uniqKey="Kobasa D" first="Darwyn" last="Kobasa">Darwyn Kobasa</name><affiliation wicri:level="1"><nlm:aff id="aff0015">High Containment Respiratory Viruses Group, Special Pathogens Program, National Microbiology Laboratory at the Canadian Science Centre for Human and Animal Health, Public Health Agency of Canada, 1015 Arlington Street, Winnipeg, Manitoba, R3E 3R2, Canada</nlm:aff><country xml:lang="fr">Canada</country><wicri:regionArea>High Containment Respiratory Viruses Group, Special Pathogens Program, National Microbiology Laboratory at the Canadian Science Centre for Human and Animal Health, Public Health Agency of Canada, 1015 Arlington Street, Winnipeg, Manitoba, R3E 3R2</wicri:regionArea><wicri:noRegion>R3E 3R2</wicri:noRegion></affiliation><affiliation wicri:level="4"><nlm:aff id="aff0025">Department of Medical Microbiology, The University of Manitoba, Winnipeg, Manitoba, R3T 2N2,Canada</nlm:aff><country xml:lang="fr">Canada</country><wicri:regionArea>Department of Medical Microbiology, The University of Manitoba, Winnipeg, Manitoba, R3T 2N2</wicri:regionArea><orgName type="university">Université du Manitoba</orgName><placeName><settlement type="city">Winnipeg</settlement><region type="state">Manitoba</region></placeName></affiliation></author><author><name sortKey="Theriault, Steven S" sort="Theriault, Steven S" uniqKey="Theriault S" first="Steven S." last="Theriault">Steven S. Theriault</name><affiliation wicri:level="1"><nlm:aff id="aff0005">Applied Biosafety Research Program, National Microbiology Laboratory at the Canadian Science Centre for Human and Animal Health, Public Health Agency of Canada, 1015 Arlington Street, Winnipeg, Manitoba, R3E 3P6, Canada</nlm:aff><country xml:lang="fr">Canada</country><wicri:regionArea>Applied Biosafety Research Program, National Microbiology Laboratory at the Canadian Science Centre for Human and Animal Health, Public Health Agency of Canada, 1015 Arlington Street, Winnipeg, Manitoba, R3E 3P6</wicri:regionArea><wicri:noRegion>R3E 3P6</wicri:noRegion></affiliation><affiliation wicri:level="1"><nlm:aff id="aff0010">National Microbiology Laboratory at the J. C. Wilt Infectious Diseases Research Centre, Public Health Agency of Canada, 745 Logan Street, Winnipeg, Manitoba, R3E 3L5, Canada</nlm:aff><country xml:lang="fr">Canada</country><wicri:regionArea>National Microbiology Laboratory at the J. C. Wilt Infectious Diseases Research Centre, Public Health Agency of Canada, 745 Logan Street, Winnipeg, Manitoba, R3E 3L5</wicri:regionArea><wicri:noRegion>R3E 3L5</wicri:noRegion></affiliation><affiliation wicri:level="4"><nlm:aff id="aff0020">Department of Microbiology, The University of Manitoba, Winnipeg, Manitoba, R3T 2N2, Canada</nlm:aff><country xml:lang="fr">Canada</country><wicri:regionArea>Department of Microbiology, The University of Manitoba, Winnipeg, Manitoba, R3T 2N2</wicri:regionArea><orgName type="university">Université du Manitoba</orgName><placeName><settlement type="city">Winnipeg</settlement><region type="state">Manitoba</region></placeName></affiliation></author></analytic><series><title level="j">Journal of Virological Methods</title><idno type="ISSN">0166-0934</idno><idno type="eISSN">1879-0984</idno><imprint><date when="2016">2016</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Animals</term><term>Cell Line</term><term>Genetic Engineering (methods)</term><term>Homologous Recombination</term><term>Middle East Respiratory Syndrome Coronavirus (genetics)</term><term>Middle East Respiratory Syndrome Coronavirus (physiology)</term><term>Reverse Genetics (methods)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Animaux</term><term>Coronavirus du syndrome respiratoire du Moyen-Orient (génétique)</term><term>Coronavirus du syndrome respiratoire du Moyen-Orient (physiologie)</term><term>Génie génétique ()</term><term>Génétique inverse ()</term><term>Lignée cellulaire</term><term>Recombinaison homologue</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Middle East Respiratory Syndrome Coronavirus</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>Coronavirus du syndrome respiratoire du Moyen-Orient</term></keywords><keywords scheme="MESH" qualifier="methods" xml:lang="en"><term>Genetic Engineering</term><term>Reverse Genetics</term></keywords><keywords scheme="MESH" qualifier="physiologie" xml:lang="fr"><term>Coronavirus du syndrome respiratoire du Moyen-Orient</term></keywords><keywords scheme="MESH" qualifier="physiology" xml:lang="en"><term>Middle East Respiratory Syndrome Coronavirus</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Animals</term><term>Cell Line</term><term>Homologous Recombination</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Animaux</term><term>Génie génétique</term><term>Génétique inverse</term><term>Lignée cellulaire</term><term>Recombinaison homologue</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><title>Highlights</title><p><list list-type="simple" id="lis0005"><list-item id="lsti0005"><label>•</label><p id="par0005">This work describes a minimalist approach to the construction and validation of a coronavirus reverse genetics system utilizing homologous recombination for vector construction and a variety of molecular techniques for virus detection.</p></list-item><list-item id="lsti0010"><label>•</label><p id="par0010">Homologous recombination in <italic>S. cerevisiae</italic> proves an efficient and fast method of constructing the large and complex genome of MERS-CoV, this strategy will be helpful for future construction of other virus genomes.</p></list-item><list-item id="lsti0015"><label>•</label><p id="par0015">Detection of rescued virus by molecular assay (i.e.: RT-PCR) is a strong alternative to the more traditional immunological detection assays.</p></list-item><list-item id="lsti0020"><label>•</label><p id="par0020">Rescued MERS-CoV was genomically and phenotypically similar to the original isolate MERS-CoV/EMC-2012.</p></list-item></list></p></div></front><back><div1 type="bibliography"><listBibl><biblStruct><analytic><author><name sortKey="Almazan, F" uniqKey="Almazan F">F. 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