Southern blot hybridization of digested deoxyribonucleic acids (DNAs) of , and with probes made of total DNA of any one of these mycoplasmas revealed hybridization bands additional to those containing ribosomal ribonucleic acid (rRNA) gene sequences, which were identifiable by specific rRNA gene probes. The number and intensity of these additional bands were most pronounced with the pair, which shares major antigenic determinants. Hybridization with and DNAs as probes revealed only the rRNA gene bands in digested DNAs of , and . DNA-DNA hybridization tests in solution, with and DNA as probes, corroborated the Southern blotting data by showing the highest homology values between and (6.5 to 8.1%), lower with (3.5 to 4.0%), and lowest with (0.5 to 0.9%). The new hybridization approach provides visual evidence for genetic relatedness between and and may facilitate the identification and cloning of genomic DNA segments carrying genes shared by two or more mycoplasmas.


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