Restriction site maps of mitochondrial DNA for Acanthamoeba strain in Korea

Abstract

The study of taxonomic relationships among members of Acanthamoeba has been based upon morphological, physiological, biochemical and immunological criteria. Studies of above methods including isoenzyme electrophoresis, however, were not consistent with various species assignments based on morphological criteria. Analysis of interstrain variability in nucleotide sequences of mitochondrial DNA(mtDNA) should be better molecular method of measuring overall genotype differences; restriction endonuclease analysis have proven to be quite powerful tool in revealing mtDNA phylogenetic relationships among closely-related organisms. In the present study, we performed the restriction endonuclease digestions of mitochondrial DNAs in order to make restriction site maps. Fragments of mtDNAs of Acanthamoeba sp. YM-4, A. culbertsoni and A. polyphaga digested with six restriction endonucleases, Cla I, Hae III, Hin dIII, Pvu II, Sal I and Spe I, have shown approximately 3-13 bands. Total mtDNA genome size by summation of six restriction enzymes averaged 42.4 kbp in Acanthamoeba sp. YM-4, 43.9 kbp in A. culbertsoni and 43.8 kbp in A. polyphaga, respectively. Fragment sizes ranged between 0.6 kbp and 39.3 kbp. Based on these data, restriction site maps were established.