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dc.contributor.authorPfefferle, Susanneen_US
dc.contributor.authorKrähling, Verenaen_US
dc.contributor.authorDitt, Vanessaen_US
dc.contributor.authorGrywna, Klausen_US
dc.contributor.authorMühlberger, Elkeen_US
dc.contributor.authorDrosten, Christianen_US
dc.date.accessioned2012-01-12T17:07:25Z
dc.date.available2012-01-12T17:07:25Z
dc.date.copyright2009
dc.date.issued2009-8-24
dc.identifier.citationPfefferle, Susanne, Verena Krähling, Vanessa Ditt, Klaus Grywna, Elke Mühlberger, Christian Drosten. "Reverse genetic characterization of the natural genomic deletion in SARS-Coronavirus strain Frankfurt-1 open reading frame 7b reveals an attenuating function of the 7b protein in-vitro and in-vivo" Virology Journal 6:131. (2009)
dc.identifier.issn1743-422X
dc.identifier.urihttps://hdl.handle.net/2144/3388
dc.description.abstractDuring the outbreak of SARS in 2002/3, a prototype virus was isolated from a patient in Frankfurt/Germany (strain Frankfurt-1). As opposed to all other SARS-Coronavirus strains, Frankfurt-1 has a 45-nucleotide deletion in the transmembrane domain of its ORF 7b protein. When over-expressed in HEK 293 cells, the full-length protein but not the variant with the deletion caused interferon beta induction and cleavage of procaspase 3. To study the role of ORF 7b in the context of virus replication, we cloned a full genome cDNA copy of Frankfurt-1 in a bacterial artificial chromosome downstream of a T7 RNA polymerase promoter. Transfection of capped RNA transcribed from this construct yielded infectious virus that was indistinguishable from the original virus isolate. The presumed Frankfurt-1 ancestor with an intact ORF 7b was reconstructed. In CaCo-2 and HUH7 cells, but not in Vero cells, the variant carrying the ORF 7b deletion had a replicative advantage against the parental virus (4- and 6-fold increase of virus RNA in supernatant, respectively). This effect was neither associated with changes in the induction or secretion of type I interferon, nor with altered induction of apoptosis in cell culture. However, pretreatment of cells with interferon beta caused the deleted virus to replicate to higher titers than the parental strain (3.4-fold in Vero cells, 7.9-fold in CaCo-2 cells). In Syrian Golden Hamsters inoculated intranasally with 10e4 plaque forming units of either virus, mean titers of infectious virus and viral RNA in the lungs after 24 h were increased 23- and 94.8-fold, respectively, with the deleted virus. This difference could explain earlier observations of enhanced virulence of Frankfurt-1 in Hamsters as compared to other SARS-Coronavirus reference strains and identifies the SARS-CoV 7b protein as an attenuating factor with the SARS-Coronavirus genome. Because attenuation was focused on the early phase of infection in-vivo, ORF 7b might have contributed to the delayed accumulation of virus in patients that was suggested to have limited the spread of the SARS epidemic.en_US
dc.description.sponsorshipGerman Ministry of Education and Research (Ökologie und Pathogenese von SARS); the European Commission (SSPE-CT-2005-022639); the German Research Foundation (Mu1365/1-1 and SFB 535); the Sino-German Center for Science Promotionen_US
dc.language.isoen
dc.publisherBioMed Centralen_US
dc.rightsCopyright 2009 Pfefferle et al; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.en_US
dc.rights.urihttp://creativecommons.org/licenses/by/2.0
dc.titleReverse Genetic Characterization of the Natural Genomic Deletion in SARS-Coronavirus Strain Frankfurt-1 Open Reading Frame 7B Reveals an Attenuating Function of the 7B Protein in-Vitro and in-Vivoen_US
dc.typeArticleen_US
dc.identifier.doi10.1186/1743-422X-6-131
dc.identifier.pmid19698190
dc.identifier.pmcid2739521


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Copyright 2009 Pfefferle et al; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Except where otherwise noted, this item's license is described as Copyright 2009 Pfefferle et al; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.