Production of infectious poliovirus from cloned cDNA is dramatically increased by SV40 transcription and replication signals.
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 plasmid 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 microgram of input DNA.
D001483:Base Sequence; D002460:Cell Line; D003001:Cloning, Molecular; D004247:DNA / Q000378:metabolism*; D004261:DNA Replication; D004262:DNA Restriction Enzymes; D004279:DNA, Viral / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics; D005814:Genes, Viral; D006801:Humans; D009690:Nucleic Acid Conformation; D010957:Plasmids; D017955:Poliovirus / Q000235:genetics*; D013539:Simian virus 40 / Q000235:genetics*; D014158:Transcription, Genetic; D014162:Transfection
D004279:DNA, Viral; D004247:DNA; D004262:DNA Restriction Enzymes