Sequence of 1060 3'-terminal nucleotides of poliovirus RNA as determined by a modification of the dideoxynucleotide method.
The dideoxynucleotide method for sequencing DNA developed by Sanger et al. [Sanger, F., Nicklen, S. & Coulson, A. (1977) Proc. Natl. Acad. Sci. USA 74, 5463-5467] was modified to allow sequence analysis of poliovirus RNA without recourse to cloning. Our method involves reverse transcription of poliovirus RNA followed by cDNA-dependent DNA synthesis in the presence of unlabeled dNTPs and 2',3'-dideoxynucleoside triphosphates, with Escherichia coli DNA polymerase I (Klenow) used to catalyze the reaction. DNA synthesis is primed by 5'-32P-labeled RNase T1- or RNase A-resistant oligonucleotides generated from poliovirus RNA. The sequence of 1060 nucleotides preceding the 3'-terminal poly(A) is presented. Based on the position of termination codons we propose that viral translation terminates at nucleotide -562.
D001483:Base Sequence; D003062:Codon / Q000032:analysis; D004247:DNA / Q000032:analysis; D004256:DNA Polymerase I; D004591:Electrophoresis, Polyacrylamide Gel; D005813:Genes, Synthetic; D017955:Poliovirus / Q000235:genetics*; D014176:Protein Biosynthesis; D012367:RNA, Viral / Q000235:genetics*; D012194:RNA-Directed DNA Polymerase
D003062:Codon; D012367:RNA, Viral; D004247:DNA; D012194:RNA-Directed DNA Polymerase; D004256:DNA Polymerase I