We have used vaccinia virus as a vector to clone a 22.5-kbp cDNA that represents the 5โ€ฒ and 3โ€ฒ 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.
year โฐ 2001
issn ๐Ÿ—„ 0022538X
volume 75
number 14
page 6676-6681
citedbycount 86