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Journal of Virology, July 2001, p. 6676-6681, Vol. 75, No. 14
0022-538X/01/$04.00+0   DOI: 10.1128/JVI.75.14.6676-6681.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Viral Replicase Gene Products Suffice for Coronavirus Discontinuous Transcription

Volker Thiel,* Jens Herold,dagger Barbara Schelle, and Stuart G. Siddell

Institute of Virology and Immunology, University of Würzburg, 97078 Würzburg, Germany

Received 28 February 2001/Accepted 23 April 2001

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.

* Corresponding author. Mailing address: Institute of Virology and Immunology, University of Würzburg, 97078 Würzburg, Germany. Phone: 49 931 201 3966. Fax: 49 931 201 3934. E-mail: v.thiel{at}mail.uni-wuerzburg.de.

dagger Present address: SWITCH-Biotech AG, 82152 Martinsried, Germany.

Journal of Virology, July 2001, p. 6676-6681, Vol. 75, No. 14
0022-538X/01/$04.00+0   DOI: 10.1128/JVI.75.14.6676-6681.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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