Dr. Murai Éva - Gubányi András szerk.: Parasitologia Hungarica 28. (Budapest, 1995)

plasmid free E. coli DH

1 to prevent contamination of the DNA samples with bacterial plasmids. Mitochondrial DNA from the isolates of Acanthamoeba was obtained by alkaline lysis. Using this method, mtDNA could be isolated from whole amoebae of monoxenic cultures without prior purification of the mitochondria. Mitochondrial DNA from the isolates was digested with two six base recognition restriction enzymes: Bgl II and EcoK I which showed the highest enzymatic activity at 37 °C (pH 7.5) and produced 6 7 fragments (Fig. 1). Fragment sizes obtained with two restriction enzymes are given in Table 2. In a comparison of our results with those reported by Yagita and Endo (1990), the RFLP phenotype of CI is comparable to JAC/E4 from 6 countries excluding Hungary, Dun is comparable to Ma from 9 countries other than Hungary, and Mos is comparable to NZAU from New Zealand (unpublished data). 0>X1

74 Lambda CI MOS Dun CI MOS Dun lambda Hae\\\Sal\ Bgl II EcoRl HindlU Fig. 1. Morphologic features of the cysts and the vegetative forms of the isolates on agar plates. A clinical isolate (Cl); B — isolate from the moss (Mos); C — isolate from the river Danube; D - Comparison of agarose gel electrophoretic patterns of mtDNA for Bgl II (lane 3: CI, lane 4: Mos, lane 5: Dun) and for EcoK 1 (lane 6: CI, lane 7: Mos, lane 8: Dun) digests. Note the usually stellated endocyst and wrinkled outer wall of cysts. The vegetative forms are larger, nuclei and granular cytoplasm can be seen. Comparing A, B and C, no morphologic distinction can be made.