Molecular cloning of poliovirus cDNA and determination of the complete nucleotide sequence of the viral genome.

TLDR

The authors synthesized double‑stranded poliovirus cDNA, inserted it into the Pst I site of plasmid pBR322, and obtained three clones that together span the entire viral genome. The complete 7410‑nt poliovirus type I genome was assembled from cloned cDNA, with two clones covering all but the first 115 bases and a third clone extending to the 5′ end, revealing an open reading frame from 743 bases downstream of the 5′ end to 71 bases upstream of the 3′ end that encodes the known poliovirus polypeptide.

Abstract

The complete 7410 nucleotide sequence of poliovirus type I genome was obtained from cloned cDNA. Double-stranded poliovirus cDNA was synthesized and inserted into the Pst I site of plasmid pBR322, and three clones were derived that together provided DNA copies of the entire poliovirus genome. Two of the clones contained inserts of 2.5 and 6.5 kilobases and represented all but the 5' 115 bases of poliovirus RNA. A third clone was generated from primer-extended DNA and contained sequences from the 5' end of the viral RNA. An open reading frame that was identified in the nucleotide sequence starting 743 bases from the 5' end of the RNA and extending to a termination codon 71 bases from the 3' end contained known poliovirus polypeptide sequence.