We examined the relationships of the genomes of five strains of Borrelia burgdorferi isolated from ticks, two from North America, including the type strain B31, and three from Switzerland. We determined restriction fragment length polymorphisms by using eight cloned DNA fragments as hybridization probes to genomic Southern blots. Two divergent patterns were observed, represented by B31 and one Swiss strain on the one hand and the two other Swiss strains on the other. The second American strain resembled B31. One of the DNA probes allowed distinction between the closely related strains within a group. The close resemblance of one Swiss strain to the North American strains suggests the possibility of their European origin. All five strains carried a circular plasmid of about 29 kb, and three contained an additional species of about 9 kb, both of which exhibited homology between the strains. The profiles of linear plasmids revealing species of 5.1 kb to 58 kb reflected the polymorphisms of chromosomal DNAs. Linear plasmids of similar size shared DNA se-quence homology. Some of the smaller plasmids tended to become lost during cultivation. © 1990, Gustav Fisher Verlag, Stuttgart · New York. All rights reserved.