Viral replicase gene products suffice for coronavirus discontinuous transcription.
We have used vaccinia virus as a vector to clone a 22.5-kbp cDNA that represents the 5' and 3' ends of the human coronavirus 229E (HCoV 229E) genome, the HCoV 229E replicase gene, and a single reporter gene (coding for green fluorescent protein [GFP]) located downstream of a regulatory element for coronavirus mRNA transcription. When RNA transcribed from this cDNA was transfected into BHK-21 cells, a small percentage of cells displayed strong fluorescence. A region of the mRNA encoding GFP was amplified by PCR and shown to have the unique mRNA leader-body junction indicative of coronavirus-mediated transcription. These data show that the coronavirus replicase gene products suffice for discontinuous subgenomic mRNA transcription.
Thiel
V
V
Institute of Virology and Immunology, University of Würzburg, 97078 Würzburg, Germany. v.thiel@mail.uni-wuerzburg.de
Herold
J
J
Schelle
B
B
Siddell
S G
SG
D017934:Coronavirus / Q000201:enzymology / Q000235:genetics*; D028941:Coronavirus 229E, Human; D005822:Genetic Vectors; D012333:RNA, Messenger / Q000235:genetics; D012367:RNA, Viral / Q000235:genetics; D012324:RNA-Dependent RNA Polymerase / Q000096:biosynthesis*; D014158:Transcription, Genetic; D014162:Transfection; D014616:Vaccinia virus / Q000235:genetics
D012333:RNA, Messenger; D012367:RNA, Viral; D012324:RNA-Dependent RNA Polymerase