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<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Synthetic recombinant bat SARS-like coronavirus is infectious in cultured cells and in mice</title><author><name sortKey="Becker, Michelle M" sort="Becker, Michelle M" uniqKey="Becker M" first="Michelle M." last="Becker">Michelle M. Becker</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Graham, Rachel L" sort="Graham, Rachel L" uniqKey="Graham R" first="Rachel L." last="Graham">Rachel L. Graham</name><affiliation><nlm:aff id="aff3"></nlm:aff></affiliation></author><author><name sortKey="Donaldson, Eric F" sort="Donaldson, Eric F" uniqKey="Donaldson E" first="Eric F." last="Donaldson">Eric F. Donaldson</name><affiliation><nlm:aff id="aff3"></nlm:aff></affiliation></author><author><name sortKey="Rockx, Barry" sort="Rockx, Barry" uniqKey="Rockx B" first="Barry" last="Rockx">Barry Rockx</name><affiliation><nlm:aff id="aff3"></nlm:aff></affiliation></author><author><name sortKey="Sims, Amy C" sort="Sims, Amy C" uniqKey="Sims A" first="Amy C." last="Sims">Amy C. Sims</name><affiliation><nlm:aff id="aff3"></nlm:aff></affiliation><affiliation><nlm:aff wicri:cut="; and" id="aff6">Carolina Vaccine Institute, University of North Carolina, Chapel Hill, NC 27599</nlm:aff></affiliation></author><author><name sortKey="Sheahan, Timothy" sort="Sheahan, Timothy" uniqKey="Sheahan T" first="Timothy" last="Sheahan">Timothy Sheahan</name><affiliation><nlm:aff id="aff3"></nlm:aff></affiliation></author><author><name sortKey="Pickles, Raymond J" sort="Pickles, Raymond J" uniqKey="Pickles R" first="Raymond J." last="Pickles">Raymond J. Pickles</name><affiliation><nlm:aff id="aff4">Microbiology and Immunology,</nlm:aff></affiliation><affiliation><nlm:aff wicri:cut=", and" id="aff5">Cystic Fibrosis/Pulmonary Research and Treatment Center and Department of Medicine</nlm:aff></affiliation></author><author><name sortKey="Corti, Davide" sort="Corti, Davide" uniqKey="Corti D" first="Davide" last="Corti">Davide Corti</name><affiliation><nlm:aff id="aff7">Institute for Research in Biomedicine, CH-6500 Bellinzona, Switzerland</nlm:aff></affiliation></author><author><name sortKey="Johnston, Robert E" sort="Johnston, Robert E" uniqKey="Johnston R" first="Robert E." last="Johnston">Robert E. Johnston</name><affiliation><nlm:aff wicri:cut="; and" id="aff6">Carolina Vaccine Institute, University of North Carolina, Chapel Hill, NC 27599</nlm:aff></affiliation></author><author><name sortKey="Baric, Ralph S" sort="Baric, Ralph S" uniqKey="Baric R" first="Ralph S." last="Baric">Ralph S. Baric</name><affiliation><nlm:aff id="aff3"></nlm:aff></affiliation><affiliation><nlm:aff wicri:cut="; and" id="aff6">Carolina Vaccine Institute, University of North Carolina, Chapel Hill, NC 27599</nlm:aff></affiliation><affiliation><nlm:aff id="aff4">Microbiology and Immunology,</nlm:aff></affiliation></author><author><name sortKey="Denison, Mark R" sort="Denison, Mark R" uniqKey="Denison M" first="Mark R." last="Denison">Mark R. Denison</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation><affiliation><nlm:aff id="aff2">Microbiology and Immunology, Vanderbilt University, Nashville, TN 37232;</nlm:aff></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">PMC</idno><idno type="pmid">19036930</idno><idno type="pmc">2588415</idno><idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2588415</idno><idno type="RBID">PMC:2588415</idno><idno type="doi">10.1073/pnas.0808116105</idno><date when="2008">2008</date><idno type="wicri:Area/Pmc/Corpus">000D14</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Corpus" wicri:corpus="PMC">000D14</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">Synthetic recombinant bat SARS-like coronavirus is infectious in cultured cells and in mice</title><author><name sortKey="Becker, Michelle M" sort="Becker, Michelle M" uniqKey="Becker M" first="Michelle M." last="Becker">Michelle M. Becker</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Graham, Rachel L" sort="Graham, Rachel L" uniqKey="Graham R" first="Rachel L." last="Graham">Rachel L. Graham</name><affiliation><nlm:aff id="aff3"></nlm:aff></affiliation></author><author><name sortKey="Donaldson, Eric F" sort="Donaldson, Eric F" uniqKey="Donaldson E" first="Eric F." last="Donaldson">Eric F. Donaldson</name><affiliation><nlm:aff id="aff3"></nlm:aff></affiliation></author><author><name sortKey="Rockx, Barry" sort="Rockx, Barry" uniqKey="Rockx B" first="Barry" last="Rockx">Barry Rockx</name><affiliation><nlm:aff id="aff3"></nlm:aff></affiliation></author><author><name sortKey="Sims, Amy C" sort="Sims, Amy C" uniqKey="Sims A" first="Amy C." last="Sims">Amy C. Sims</name><affiliation><nlm:aff id="aff3"></nlm:aff></affiliation><affiliation><nlm:aff wicri:cut="; and" id="aff6">Carolina Vaccine Institute, University of North Carolina, Chapel Hill, NC 27599</nlm:aff></affiliation></author><author><name sortKey="Sheahan, Timothy" sort="Sheahan, Timothy" uniqKey="Sheahan T" first="Timothy" last="Sheahan">Timothy Sheahan</name><affiliation><nlm:aff id="aff3"></nlm:aff></affiliation></author><author><name sortKey="Pickles, Raymond J" sort="Pickles, Raymond J" uniqKey="Pickles R" first="Raymond J." last="Pickles">Raymond J. Pickles</name><affiliation><nlm:aff id="aff4">Microbiology and Immunology,</nlm:aff></affiliation><affiliation><nlm:aff wicri:cut=", and" id="aff5">Cystic Fibrosis/Pulmonary Research and Treatment Center and Department of Medicine</nlm:aff></affiliation></author><author><name sortKey="Corti, Davide" sort="Corti, Davide" uniqKey="Corti D" first="Davide" last="Corti">Davide Corti</name><affiliation><nlm:aff id="aff7">Institute for Research in Biomedicine, CH-6500 Bellinzona, Switzerland</nlm:aff></affiliation></author><author><name sortKey="Johnston, Robert E" sort="Johnston, Robert E" uniqKey="Johnston R" first="Robert E." last="Johnston">Robert E. Johnston</name><affiliation><nlm:aff wicri:cut="; and" id="aff6">Carolina Vaccine Institute, University of North Carolina, Chapel Hill, NC 27599</nlm:aff></affiliation></author><author><name sortKey="Baric, Ralph S" sort="Baric, Ralph S" uniqKey="Baric R" first="Ralph S." last="Baric">Ralph S. Baric</name><affiliation><nlm:aff id="aff3"></nlm:aff></affiliation><affiliation><nlm:aff wicri:cut="; and" id="aff6">Carolina Vaccine Institute, University of North Carolina, Chapel Hill, NC 27599</nlm:aff></affiliation><affiliation><nlm:aff id="aff4">Microbiology and Immunology,</nlm:aff></affiliation></author><author><name sortKey="Denison, Mark R" sort="Denison, Mark R" uniqKey="Denison M" first="Mark R." last="Denison">Mark R. Denison</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation><affiliation><nlm:aff id="aff2">Microbiology and Immunology, Vanderbilt University, Nashville, TN 37232;</nlm:aff></affiliation></author></analytic><series><title level="j">Proceedings of the National Academy of Sciences of the United States of America</title><idno type="ISSN">0027-8424</idno><idno type="eISSN">1091-6490</idno><imprint><date when="2008">2008</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p>Defining prospective pathways by which zoonoses evolve and emerge as human pathogens is critical for anticipating and controlling both natural and deliberate pandemics. However, predicting tenable pathways of animal-to-human movement has been hindered by challenges in identifying reservoir species, cultivating zoonotic organisms in culture, and isolating full-length genomes for cloning and genetic studies. The ability to design and recover pathogens reconstituted from synthesized cDNAs has the potential to overcome these obstacles by allowing studies of replication and pathogenesis without identification of reservoir species or cultivation of primary isolates. Here, we report the design, synthesis, and recovery of the largest synthetic replicating life form, a 29.7-kb bat severe acute respiratory syndrome (SARS)-like coronavirus (Bat-SCoV), a likely progenitor to the SARS-CoV epidemic. To test a possible route of emergence from the noncultivable Bat-SCoV to human SARS-CoV, we designed a consensus Bat-SCoV genome and replaced the Bat-SCoV Spike receptor-binding domain (RBD) with the SARS-CoV RBD (Bat-SRBD). Bat-SRBD was infectious in cell culture and in mice and was efficiently neutralized by antibodies specific for both bat and human CoV Spike proteins. Rational design, synthesis, and recovery of hypothetical recombinant viruses can be used to investigate mechanisms of transspecies movement of zoonoses and has great potential to aid in rapid public health responses to known or predicted emerging microbial threats.</p></div></front></TEI><pmc article-type="research-article"><pmc-comment>The publisher of this article does not allow downloading of the full text in XML form.</pmc-comment>
<front><journal-meta><journal-id journal-id-type="nlm-ta">Proc Natl Acad Sci U S A</journal-id><journal-id journal-id-type="hwp">pnas</journal-id><journal-id journal-id-type="pmc">pnas</journal-id><journal-id journal-id-type="publisher-id">PNAS</journal-id><journal-title-group><journal-title>Proceedings of the National Academy of Sciences of the United States of America</journal-title></journal-title-group><issn pub-type="ppub">0027-8424</issn><issn pub-type="epub">1091-6490</issn><publisher><publisher-name>National Academy of Sciences</publisher-name></publisher></journal-meta><article-meta><article-id pub-id-type="pmid">19036930</article-id><article-id pub-id-type="pmc">2588415</article-id><article-id pub-id-type="publisher-id">5757</article-id><article-id pub-id-type="doi">10.1073/pnas.0808116105</article-id><article-categories><subj-group subj-group-type="heading"><subject>Biological Sciences</subject><subj-group><subject>Microbiology</subject></subj-group></subj-group></article-categories><title-group><article-title>Synthetic recombinant bat SARS-like coronavirus is infectious in cultured cells and in mice</article-title></title-group><contrib-group><contrib contrib-type="author"><name><surname>Becker</surname><given-names>Michelle M.</given-names></name><xref ref-type="aff" rid="aff1"><sup>a</sup></xref><xref ref-type="author-notes" rid="FN1"><sup>1</sup></xref></contrib><contrib contrib-type="author"><name><surname>Graham</surname><given-names>Rachel L.</given-names></name><xref ref-type="aff" rid="aff3"><sup>b</sup></xref><xref ref-type="author-notes" rid="FN1"><sup>1</sup></xref></contrib><contrib contrib-type="author"><name><surname>Donaldson</surname><given-names>Eric F.</given-names></name><xref ref-type="aff" rid="aff3"><sup>b</sup></xref></contrib><contrib contrib-type="author"><name><surname>Rockx</surname><given-names>Barry</given-names></name><xref ref-type="aff" rid="aff3"><sup>b</sup></xref></contrib><contrib contrib-type="author"><name><surname>Sims</surname><given-names>Amy C.</given-names></name><xref ref-type="aff" rid="aff3"><sup>b</sup></xref><xref ref-type="aff" rid="aff6"><sup>c</sup></xref></contrib><contrib contrib-type="author"><name><surname>Sheahan</surname><given-names>Timothy</given-names></name><xref ref-type="aff" rid="aff3"><sup>b</sup></xref></contrib><contrib contrib-type="author"><name><surname>Pickles</surname><given-names>Raymond J.</given-names></name><xref ref-type="aff" rid="aff4"><sup>d</sup></xref><xref ref-type="aff" rid="aff5"><sup>e</sup></xref></contrib><contrib contrib-type="author"><name><surname>Corti</surname><given-names>Davide</given-names></name><xref ref-type="aff" rid="aff7"><sup>f</sup></xref></contrib><contrib contrib-type="author"><name><surname>Johnston</surname><given-names>Robert E.</given-names></name><xref ref-type="aff" rid="aff6"><sup>c</sup></xref></contrib><contrib contrib-type="author"><name><surname>Baric</surname><given-names>Ralph S.</given-names></name><xref ref-type="aff" rid="aff3"><sup>b</sup></xref><xref ref-type="aff" rid="aff6"><sup>c</sup></xref><xref ref-type="aff" rid="aff4"><sup>d</sup></xref><xref ref-type="corresp" rid="cor1"><sup>2</sup></xref></contrib><contrib contrib-type="author"><name><surname>Denison</surname><given-names>Mark R.</given-names></name><xref ref-type="aff" rid="aff1"><sup>a</sup></xref><xref ref-type="aff" rid="aff2"><sup>g</sup></xref><xref ref-type="corresp" rid="cor1"><sup>2</sup></xref></contrib><aff id="aff1">Departments of<sup>a</sup>Pediatrics and</aff><aff id="aff2"><sup>g</sup>Microbiology and Immunology, Vanderbilt University, Nashville, TN 37232;</aff><aff id="aff3">Departments of<sup>b</sup>Epidemiology and</aff><aff id="aff4"><sup>d</sup>Microbiology and Immunology,</aff><aff id="aff5"><sup>e</sup>Cystic Fibrosis/Pulmonary Research and Treatment Center and Department of Medicine, and</aff><aff id="aff6"><sup>c</sup>Carolina Vaccine Institute, University of North Carolina, Chapel Hill, NC 27599; and</aff><aff id="aff7"><sup>f</sup>Institute for Research in Biomedicine, CH-6500 Bellinzona, Switzerland</aff></contrib-group><author-notes><corresp id="cor1"><sup>2</sup>To whom correspondence may be addressed. E-mail: <email>rbaric@email.unc.edu</email> or <email>mark.denison@vanderbilt.edu</email></corresp><fn fn-type="edited-by"><p>Edited by Peter Palese, Mount Sinai School of Medicine, New York, NY, and approved October 14, 2008</p></fn><fn fn-type="con"><p>Author contributions: M.M.B., R.L.G., E.F.D., R.S.B., and M.R.D. designed research; M.M.B., R.L.G., E.F.D., B.R., and A.C.S. performed research; R.L.G., E.F.D., T.S., R.J.P., D.C., and R.E.J. contributed new reagents/analytic tools; M.M.B., R.L.G., E.F.D., B.R., A.C.S., R.J.P., and R.S.B. analyzed data; and M.M.B., R.L.G., R.S.B., and M.R.D. wrote the paper.</p></fn><fn fn-type="equal" id="FN1"><p><sup>1</sup>M.M.B and R.L.G. contributed equally to this work.</p></fn><fn fn-type="conflict"><p>Conflict of interest statement: R.E.J. is a coinventor of the Venezuelan Equine Encephalitis (VEE) expression vector technology and holds an equity interest in AlphaVax, Inc., the company that has licensed this technology from the University of North Carolina.</p></fn></author-notes><pub-date pub-type="ppub"><day>16</day><month>12</month><year>2008</year></pub-date><pub-date pub-type="epub"><day>26</day><month>11</month><year>2008</year></pub-date><pub-date pub-type="pmc-release"><day>26</day><month>11</month><year>2008</year></pub-date><pmc-comment> PMC Release delay is 0 months and 0 days and was based on the
epub date downloaded from Highwire. </pmc-comment>
<volume>105</volume><issue>50</issue><fpage>19944</fpage><lpage>19949</lpage><history><date date-type="received"><day>18</day><month>8</month><year>2008</year></date></history><permissions><copyright-statement>© 2008 by The National Academy of Sciences of the USA</copyright-statement><license license-type="open-access"><license-p>Freely available online through the PNAS open access option.</license-p></license></permissions><self-uri xlink:title="pdf" xlink:type="simple" xlink:href="zpq05008019944.pdf"></self-uri><abstract><p>Defining prospective pathways by which zoonoses evolve and emerge as human pathogens is critical for anticipating and controlling both natural and deliberate pandemics. However, predicting tenable pathways of animal-to-human movement has been hindered by challenges in identifying reservoir species, cultivating zoonotic organisms in culture, and isolating full-length genomes for cloning and genetic studies. The ability to design and recover pathogens reconstituted from synthesized cDNAs has the potential to overcome these obstacles by allowing studies of replication and pathogenesis without identification of reservoir species or cultivation of primary isolates. Here, we report the design, synthesis, and recovery of the largest synthetic replicating life form, a 29.7-kb bat severe acute respiratory syndrome (SARS)-like coronavirus (Bat-SCoV), a likely progenitor to the SARS-CoV epidemic. To test a possible route of emergence from the noncultivable Bat-SCoV to human SARS-CoV, we designed a consensus Bat-SCoV genome and replaced the Bat-SCoV Spike receptor-binding domain (RBD) with the SARS-CoV RBD (Bat-SRBD). Bat-SRBD was infectious in cell culture and in mice and was efficiently neutralized by antibodies specific for both bat and human CoV Spike proteins. Rational design, synthesis, and recovery of hypothetical recombinant viruses can be used to investigate mechanisms of transspecies movement of zoonoses and has great potential to aid in rapid public health responses to known or predicted emerging microbial threats.</p></abstract><kwd-group><kwd>emerging pathogens</kwd><kwd>synthetic biology</kwd><kwd>vaccine development</kwd><kwd>zoonoses</kwd></kwd-group></article-meta></front></pmc></record>Pour manipuler ce document sous Unix (Dilib)
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