Rapid one-step construction of a Middle East Respiratory Syndrome (MERS-CoV) infectious clone system by homologous recombination
Identifieur interne : 000A23 ( Pmc/Checkpoint ); précédent : 000A22; suivant : 000A24Rapid 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.
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
Affiliations:
Links toward previous steps (curation, corpus...)
Links to Exploration step
PMC:7113859Le document en format XML
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<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>
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<pmc article-type="research-article"><pmc-dir>properties open_access</pmc-dir>
<front><journal-meta><journal-id journal-id-type="nlm-ta">J Virol Methods</journal-id>
<journal-id journal-id-type="iso-abbrev">J. Virol. Methods</journal-id>
<journal-title-group><journal-title>Journal of Virological Methods</journal-title>
</journal-title-group>
<issn pub-type="ppub">0166-0934</issn>
<issn pub-type="epub">1879-0984</issn>
<publisher><publisher-name>Published by Elsevier B.V.</publisher-name>
</publisher>
</journal-meta>
<article-meta><article-id pub-id-type="pmid">27459876</article-id>
<article-id pub-id-type="pmc">7113859</article-id>
<article-id pub-id-type="publisher-id">S0166-0934(16)30312-3</article-id>
<article-id pub-id-type="doi">10.1016/j.jviromet.2016.07.022</article-id>
<article-categories><subj-group subj-group-type="heading"><subject>Article</subject>
</subj-group>
</article-categories>
<title-group><article-title>Rapid one-step construction of a Middle East Respiratory Syndrome (MERS-CoV) infectious clone system by homologous recombination</article-title>
</title-group>
<contrib-group><contrib contrib-type="author" id="aut0005"><name><surname>Nikiforuk</surname>
<given-names>Aidan M.</given-names>
</name>
<xref rid="aff0005" ref-type="aff">a</xref>
<xref rid="aff0010" ref-type="aff">b</xref>
<xref rid="aff0015" ref-type="aff">c</xref>
<xref rid="aff0020" ref-type="aff">d</xref>
</contrib>
<contrib contrib-type="author" id="aut0010"><name><surname>Leung</surname>
<given-names>Anders</given-names>
</name>
<xref rid="aff0015" ref-type="aff">c</xref>
</contrib>
<contrib contrib-type="author" id="aut0015"><name><surname>Cook</surname>
<given-names>Bradley W.M.</given-names>
</name>
<xref rid="aff0005" ref-type="aff">a</xref>
<xref rid="aff0010" ref-type="aff">b</xref>
<xref rid="aff0020" ref-type="aff">d</xref>
</contrib>
<contrib contrib-type="author" id="aut0020"><name><surname>Court</surname>
<given-names>Deborah A.</given-names>
</name>
<xref rid="aff0020" ref-type="aff">d</xref>
</contrib>
<contrib contrib-type="author" id="aut0025"><name><surname>Kobasa</surname>
<given-names>Darwyn</given-names>
</name>
<xref rid="aff0015" ref-type="aff">c</xref>
<xref rid="aff0025" ref-type="aff">e</xref>
</contrib>
<contrib contrib-type="author" id="aut0030"><name><surname>Theriault</surname>
<given-names>Steven S.</given-names>
</name>
<email>steven.theriault@phac-aspc.gc.ca</email>
<xref rid="aff0005" ref-type="aff">a</xref>
<xref rid="aff0010" ref-type="aff">b</xref>
<xref rid="aff0020" ref-type="aff">d</xref>
<xref rid="cor0005" ref-type="corresp">⁎</xref>
</contrib>
</contrib-group>
<aff id="aff0005"><label>a</label>
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</aff>
<aff id="aff0010"><label>b</label>
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</aff>
<aff id="aff0015"><label>c</label>
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</aff>
<aff id="aff0020"><label>d</label>
Department of Microbiology, The University of Manitoba, Winnipeg, Manitoba, R3T 2N2, Canada</aff>
<aff id="aff0025"><label>e</label>
Department of Medical Microbiology, The University of Manitoba, Winnipeg, Manitoba, R3T 2N2,Canada</aff>
<author-notes><corresp id="cor0005"><label>⁎</label>
Corresponding author at: 1015 Arlington Street, Winnipeg, Manitoba, R3E 3P6, Canada. <email>steven.theriault@phac-aspc.gc.ca</email>
</corresp>
</author-notes>
<pub-date pub-type="pmc-release"><day>25</day>
<month>7</month>
<year>2016</year>
</pub-date>
<pmc-comment> PMC Release delay is 0 months and 0 days and was based on .</pmc-comment>
<pub-date pub-type="ppub"><month>10</month>
<year>2016</year>
</pub-date>
<pub-date pub-type="epub"><day>25</day>
<month>7</month>
<year>2016</year>
</pub-date>
<volume>236</volume>
<fpage>178</fpage>
<lpage>183</lpage>
<history><date date-type="received"><day>14</day>
<month>6</month>
<year>2016</year>
</date>
<date date-type="rev-recd"><day>21</day>
<month>7</month>
<year>2016</year>
</date>
<date date-type="accepted"><day>22</day>
<month>7</month>
<year>2016</year>
</date>
</history>
<permissions><copyright-statement>Crown Copyright © 2016 Published by Elsevier B.V.</copyright-statement>
<copyright-year>2016</copyright-year>
<copyright-holder></copyright-holder>
<license><license-p>Since January 2020 Elsevier has created a COVID-19 resource centre with free information in English and Mandarin on the novel coronavirus COVID-19. The COVID-19 resource centre is hosted on Elsevier Connect, the company's public news and information website. Elsevier hereby grants permission to make all its COVID-19-related research that is available on the COVID-19 resource centre - including this research content - immediately available in PubMed Central and other publicly funded repositories, such as the WHO COVID database with rights for unrestricted research re-use and analyses in any form or by any means with acknowledgement of the original source. These permissions are granted for free by Elsevier for as long as the COVID-19 resource centre remains active.</license-p>
</license>
</permissions>
<abstract abstract-type="author-highlights" id="abs0005"><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>
</abstract>
<abstract id="abs0010"><sec><title>Background</title>
<p>Viral Infectious clone systems serve as robust platforms to study viral gene or replicative function by reverse genetics, formulate vaccines and adapt a wild type-virus to an animal host. Since the development of the first viral infectious clone system for the poliovirus, novel strategies of viral genome construction have allowed for the assembly of viral genomes across the identified viral families. However, the molecular profiles of some viruses make their genome more difficult to construct than others. Two factors that affect the difficulty of infectious clone construction are genome length and genome complexity.</p>
</sec>
<sec><title>Results</title>
<p>This work examines the available strategies for overcoming the obstacles of assembling the long and complex RNA genomes of coronaviruses and reports one-step construction of an infectious clone system for the Middle East Respiratory Syndrome coronavirus (MERS-CoV) by homologous recombination in <italic>S. cerevisiae</italic>
.</p>
</sec>
<sec><title>Conclusions</title>
<p>Future use of this methodology will shorten the time between emergence of a novel viral pathogen and construction of an infectious clone system. Completion of a viral infectious clone system facilitates further study of a virus’s biology, improvement of diagnostic tests, vaccine production and the screening of antiviral compounds.</p>
</sec>
</abstract>
<kwd-group id="kwd0005"><title>Keywords</title>
<kwd>Coronavirus</kwd>
<kwd>Middle East Respiratory Syndrome</kwd>
<kwd>Infectious clone system</kwd>
<kwd>Molecular cloning</kwd>
<kwd>Reverse genetics</kwd>
<kwd>Homologous recombination</kwd>
</kwd-group>
</article-meta>
</front>
</pmc>
<affiliations><list><country><li>Canada</li>
</country>
<region><li>Manitoba</li>
</region>
<settlement><li>Winnipeg</li>
</settlement>
<orgName><li>Université du Manitoba</li>
</orgName>
</list>
<tree><country name="Canada"><noRegion><name sortKey="Nikiforuk, Aidan M" sort="Nikiforuk, Aidan M" uniqKey="Nikiforuk A" first="Aidan M." last="Nikiforuk">Aidan M. Nikiforuk</name>
</noRegion>
<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>
<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>
<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>
<name sortKey="Court, Deborah A" sort="Court, Deborah A" uniqKey="Court D" first="Deborah A." last="Court">Deborah A. Court</name>
<name sortKey="Kobasa, Darwyn" sort="Kobasa, Darwyn" uniqKey="Kobasa D" first="Darwyn" last="Kobasa">Darwyn Kobasa</name>
<name sortKey="Kobasa, Darwyn" sort="Kobasa, Darwyn" uniqKey="Kobasa D" first="Darwyn" last="Kobasa">Darwyn Kobasa</name>
<name sortKey="Leung, Anders" sort="Leung, Anders" uniqKey="Leung A" first="Anders" last="Leung">Anders Leung</name>
<name sortKey="Nikiforuk, Aidan M" sort="Nikiforuk, Aidan M" uniqKey="Nikiforuk A" first="Aidan M." last="Nikiforuk">Aidan M. Nikiforuk</name>
<name sortKey="Nikiforuk, Aidan M" sort="Nikiforuk, Aidan M" uniqKey="Nikiforuk A" first="Aidan M." last="Nikiforuk">Aidan M. Nikiforuk</name>
<name sortKey="Nikiforuk, Aidan M" sort="Nikiforuk, Aidan M" uniqKey="Nikiforuk A" first="Aidan M." last="Nikiforuk">Aidan M. Nikiforuk</name>
<name sortKey="Theriault, Steven S" sort="Theriault, Steven S" uniqKey="Theriault S" first="Steven S." last="Theriault">Steven S. Theriault</name>
<name sortKey="Theriault, Steven S" sort="Theriault, Steven S" uniqKey="Theriault S" first="Steven S." last="Theriault">Steven S. Theriault</name>
<name sortKey="Theriault, Steven S" sort="Theriault, Steven S" uniqKey="Theriault S" first="Steven S." last="Theriault">Steven S. Theriault</name>
</country>
</tree>
</affiliations>
</record>
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