Reverse genetics with a full-length infectious cDNA of severe acute respiratory syndrome coronavirus.
A previously undescribed coronavirus (CoV) is the etiologic agent responsible for severe acute respiratory syndrome (SARS). Using a panel of contiguous cDNAs that span the entire genome, we have assembled a full-length cDNA of the SARS-CoV Urbani strain, and have rescued molecularly cloned SARS viruses (infectious clone SARS-CoV) that contained the expected marker mutations inserted into the component clones. Recombinant viruses replicated as efficiently as WT virus and both were inhibited by treatment with the cysteine proteinase inhibitor (2S,3S)-transepoxysuccinyl-L-leucylamido-3-methylbutane ethyl ester. In addition, subgenomic transcripts were initiated from the consensus sequence ACGAAC in both the WT and infectious clone SARS-CoV. Availability of a SARS-CoV full-length cDNA provides a template for manipulation of the viral genome, allowing for the rapid and rational development and testing of candidate vaccines and therapeutics against this important human pathogen.
Department of Epidemiology and Microbiology, University of North Carolina, Chapel Hill, NC 27599-7435, USA.
Boyd
B
Yount
Kristopher M
KM
Curtis
Elizabeth A
EA
Fritz
Lisa E
LE
Hensley
Peter B
PB
Jahrling
Erik
E
Prentice
Mark R
MR
Denison
Thomas W
TW
Geisbert
Ralph S
RS
Baric
D000818:Animals; D001483:Base Sequence; D002522:Chlorocebus aethiops; D003001:Cloning, Molecular; D016384:Consensus Sequence; D018076:DNA, Complementary / Q000235:genetics*; D005819:Genetic Markers; D005821:Genetic Techniques; D006801:Humans; D016366:Open Reading Frames; D045473:Severe acute respiratory syndrome-related coronavirus / Q000235:genetics* / Q000472:pathogenicity; D045169:Severe Acute Respiratory Syndrome / Q000821:virology; D014158:Transcription, Genetic; D014709:Vero Cells
D018076:DNA, Complementary; D005819:Genetic Markers