The complete nucleotide sequences of the genomes of the type 2 (P712, Ch, 2ab) and type 3 (Leon 12a₁b) poliovirus vaccine strains were determined. Comparison of the sequences with the previously established genome sequence of type 1 (LS-c, 2ab) poliovirus vaccine strain revealed that 71% of the nucleotides in the genome RNAs were common, that the 5′ and 3′ termini of the genomes were highly homologous, and that more than 80% of the nucleotide differences in the coding region occurred in the third letter position of in-phase codons, resulting in a low frequency of amino acid difference. These results strongly suggested that the serotypes of poliovirus derived from a common prototype. A comparison of the amino acid sequences predicted from the genome sequences showed highest variation in the capsid protein region, whereas non-structural proteins are highly conserved. Initiation of polyprotein synthesis occurs in all three strains more than 740 nucleotides downstream from the 5′ end. An analysis of the non-coding region suggests that small peptides that could potentially originate from this region are not involved in viral replication. All cleavage sites in polyprotein processing are conserved. The amino acid sequences immediately surrounding the cleavage signals, however, show a higher than average degree of variation. The analysis of the amino acid sequences of the capsid protein VP1 of all serotypes has led to the prediction of potential antigenic sites on the virion involved in neutralization.