Purified deoxyribonucleic acids (DNAs) of representative strains of nine established serovars of the human mycoplasma and of five strains of were digested with a variety of restriction endonucleases. The cleavage patterns obtained by electrophoresis of the digestion products separated the nine serovars into two definite clusters, one consisting of serovars 1, 3, and 6 and the other consisting of serovars 2, 4, 5, 7, 8, and 9, in agreement with previous results obtained by electrophoresis of cell proteins and DNA-DNA hybridization. Although the five strains differed in virulence and adherence capacity, they exhibited similar DNA cleavage patterns, indicating a high degree of genetic homogeneity. The enzymes HI, and I, which possess guanine-plus-cytosine-rich recognition sequences, cleaved the DNA of (guanine-plus-cytosine content, about 28 mol%) into a small number of fragments, whereas I, with the recognition sequence CCC/GGG, failed to cleave DNA into visibly distinct fragments. The same restriction enzymes produced multiband cleavage patterns with DNA, which has a guanine-plus-cytosine content of about 40 mol%. Restriction endonucleases RI, III, I, and XI, which have recognition sequences rich in adenine plus thymine, produced multiband patterns with the DNAs of both and . We concluded that the cleavage patterns of mycoplasmal DNAs digested with restriction endonucleases provide a means for determining genetic relatedness among mycoplasmas.


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