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Journal of Virology, November 1998, p. 9365-9369, Vol. 72, No. 11
0022-538X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Identification and Characterization of an RNA-Dependent RNA Polymerase Activity within the Nonstructural Protein 5B Region of Bovine Viral Diarrhea Virus

Weidong Zhong,* Lester L. Gutshall, and Alfred M. Del Vecchio

Department of Molecular Virology and Host Defense, SmithKline Beecham Pharmaceuticals, Collegeville, Pennsylvania 19426

Received 30 March 1998/Accepted 24 July 1998

Nonstructural protein 5B (NS5B) of bovine viral diarrhea virus (BVDV) contains sequence motifs that are predictive of an RNA-dependent RNA polymerase activity. We describe the expression and purification of the BVDV NS5B protein derived from an infectious cDNA clone of BVDV (NADL strain). BVDV NS5B protein was active in an in vitro RNA polymerase assay using homopolymeric RNA or BVDV minigenomic RNA templates. The major product was a covalently linked double-stranded molecule generated by a "copy-back" mechanism from the input template RNA. In addition, a nucleotide-nonspecific and template-independent terminal nucleotidyl transferase activity was observed with the BVDV NS5B preparation.

* Corresponding author. Mailing address: Department of Molecular Virology and Host Defense, SmithKline Beecham Pharmaceuticals, 1250 S. Collegeville Rd., P.O. Box 5089, Collegeville, PA 19426-0989. Phone: (610) 917-6356. Fax: (610) 917-4170. E-mail: weidong_zhong-1{at}sbphrd.com.

Journal of Virology, November 1998, p. 9365-9369, Vol. 72, No. 11
0022-538X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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