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Journal of Virology, July 2000, p. 6394-6400, Vol. 74, No. 14
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Poliovirus Requires a Precise 5' End for Efficient Positive-Strand RNA Synthesis

Jens Herold and Raul Andino*

Department of Microbiology and Immunology, University of California at San Francisco, San Francisco, California 94143-0414

Received 21 November 1999/Accepted 24 April 2000

Poliovirus infectious RNA can be synthesized in vitro using phage DNA-dependent RNA-polymerases. These synthetic transcripts contain several extra nucleotides at the 5' end, which are deleted during replication to generate authentic viral genomes. We removed those 5'-end extra nucleotides utilizing a hammerhead ribozyme to produce transcripts with accurate 5' ends. These transcripts replicate substantially more rapidly in cell culture, demonstrating no lag before replication; they also replicate more efficiently in Xenopus laevis oocytes and in in vitro translation-replication cell extracts. In both systems, an exact 5' end is necessary for synthesis of positive-strand RNA but not negative-strand RNA.

* Corresponding author. Mailing address: University of California at San Francisco, Department of Microbiology and Immunology, 513 Parnassus Ave., Box 0414, San Francisco, CA 94143-0414. Phone: (415) 502-6358. Fax: (415) 476-0939. E-mail: andino{at}cgl.ucsf.edu.

Journal of Virology, July 2000, p. 6394-6400, Vol. 74, No. 14
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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