The typing bacteriophages 55, 80, 83A, and 85 of Staphylococcus aureus, representative of the three major lytic groups of serological group B aureophages, have been examined for relatedness of their genomes and virion proteins. Phages 11 and 80a were also examined to determine the relationship of phage 80a to phages 11 and 80. Total genome hybridization measurements divided the phages into two groups. Phages 55 and 80, in the first group, had DNA homology of 50%. Phages 11, 80a, 83A, and 85 formed a second group with 27 to 65% homology. Homology between the two groups was in the range of 14 to 22%. Phage 80a is more closely related to phage 11 than to phage 80, though it is probably not a simple recombinant of phages 11 and 80. Restriction enzyme digestion and phage [32PJDNA hybridization analysis of the endonuclease-generated fragments from each phage DNA confirmed the findings of the DNA homology measurements. The endonuclease fragment patterns generated by EcoRI and HindlIl were distinctive for eacb phage, confirming that none of the phages are closely related. Common sequences were present in most fragments from the phage DNAs when the labeled probe DNA was from a different phage in the same group. Cross-group probing of endonuclease fragments revealed both a diminished level of homology when similar sequences were present and the probable absence of some sequences. Virion proteins, examined by polyacrylamide gel electrophoresis, were similar in number and molecular weight for phages 11, 80a, 83A, and 85, reflecting the DNA homology analyses. The virion proteins from phages 55 and 80, however, were more distinctive, and both differed from the phages in the other group.

Fifteen bacteriophages representative of the serological and lytic groups of the International Typing Set for Staphylococcus aureus were examined for genomic homology by DNA hybridization and by analysis of high resolution thermal denaturation profiles. Phages 11 and 80 alpha, not part of the set, were also examined. DNA homology measured by filter hybridization showed values ranging from near zero to 88 per cent in pair-wise comparisons. Cluster analysis of these data by standard numerical taxonomical methods yielded clusters which closely reflect the subdivision of the international set on the basis of serological reactions. High resolution thermal denaturation analysis yielded characteristic profiles for each phage DNA, with members closely related by hybridization analysis showing only minor differences. Quantitative analysis of the extent of overlap of these profiles generated relational values which were subjected to the same numerical taxonomic analysis as for the DNA hybridization data. The resultant dendrogram was qualitatively different only in minor respects from that derived from the hybridization analyses, but quantitatively homology was greater by 80 per cent or more for the DNAs which were least related according to the hybridization analyses. This upward shift in measured homology appears to reflect the similar base composition of the DNAs from these phages. Statistical comparison of the homology data obtained by the two methods showed them to be significantly correlated. These results indicate that the International Typing Set consists of phages which all appear to be related to a greater or lesser extent. If, as the history of the collection of these phages indicates, they are a random sample of aureophages, then this group of phages may represent a common genetic pool within which recombination, mutation, and genome rearrangement occur to generate unique individual phages.