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Nucleic Acids Research, 1984, Vol. 12, No. 12 5123-5141
© 1984

MOLECULAR BIOLOGY

Production of infectious poliovirus from cloned cDNA is dramatically increased by SV40 transcription and replication signals

Ben L. Semler, Andrew J. Dorner*,+ and Eckard Wimmer*

Department of Microbiology and Molecular Genetics, College of Medicine, University of California Irvine, CA 92717 *Department of Microbiology, School of Medicine, State University of New York Stonv Brook. NY 11794. USA

Received February 14, 1984. Revised May 25, 1984. Accepted May 25, 1984.

Sub-genomic cDNA clones representing the entire genomic RNA of poliovirus Type 1 (Mahoney) have been isolated in E. coli. Construction of a complete cDNA copy of the poliovirus genome in the EcoRI site of plasm id vector pBR325 from these clones is described. Introduction of plasmid DNA containing the complete cDNA copy of polio RNA into cultured primate cells by transfection produces infectious poliovirus. The virus produced by such a transfection appears to be identical to wild type poliovirus. Isolation of a polio recombinant plasmid containing SV40 transcription and replication signals is also described. Transfection of COS-1 cells with this plasmid yields greater than 1,600 plaque-forming units (PFU) per ug of input DNA.

+Present address: Department of Molecular Biology and Microbiology, Tufts University School of Medicine, Boston, MA 02111, USA


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