Restriction endonuclease analysis of mitochondrial DNA of <i>Acanthamoeba</i> sp. YM-4 (Korean isolate)

, Shin, H J; , Im, K; , Jeon, K W

doi:1997.35.2.119

Korean J Parasito > Volume 35(2):1997 > Article

Original Article


Korean J Parasitol. 1997 Jun;35(2):119-125. English. Published online Jun 20, 1997. http://dx.doi.org/10.3347/kjp.1997.35.2.119
Copyright © 1997 by The Korean Society for Parasitology


Restriction endonuclease analysis of mitochondrial DNA of Acanthamoeba sp. YM-4 (Korean isolate)
H J Shin,^*¹K Im,² and K W Jeon³
¹Department of Microbiology, Ajou University School of Medicine, Suwon 442-749, Korea.

Received February 11, 1997; Accepted May 28, 1997.
Abstract
Acanthamoeba sp. YM-4 is similar to A. culbertsoni based upon morphological characteristics of trophozoites and cysts. However, based on other characteristics, pathogenicity to mice, in vitro cytotoxicity and isoenzyme patterns. Acanthamoeba sp. YM-4 was quite different from A. culbertsoni. Restriction fragment length polymorphism (RFLP) analysis of mtDNA is useful in the classification of members belonging to the genus Acanthamoeba. Therefore, in this study, RFLP analysis of Acanthamoeba mtDNAs was accomplished using five restriction enzymes: HaeIII, HindIII, ClaI, PvuII and SalI. Each restriction enzyme produced approximately 3-15 fragments (range: from 0.6 kbp to 34.4 kbp). The mtDNA genome size, calculated by the summation of restriction fragments, averaged 46.4 kbp in Acanthamoeba sp. YM-4, 48.3 kbp in A. culbertsoni and 48.8 kbp in A. polyphaga, respectively. Digested mtDNA fragments of Acanthamoeba sp. YM-4 contained nine and seven same size fragments, respectively, from a total of 67 and 69 fragments observed in A. culbertsoni and A. polyphaga. An estimate of the genetic divergence was 10.1% between Acanthamoeba sp. YM-4 and A. culbertsoni, and 9.9% between Acanthamoeba sp. YM-4 and A. polyphaga.

Figures


	Fig. 1 DNA gel electrophoretic patterns of Acanthamoeba YM-4 (lane 2), A. culbertsoni (lane 3) and A. polyphaga (lane 4) after digestion with HaeIII, HindIII, ClaI, PvuII and SalI, respectively. Lane 1 is the size marker (λ DNA digested with HindIII)


	Fig. 2 Dendrogram showing estimated genetic divergence among Acanthamoeba spp. using UPGMA cluster analysis.

Tables


	Table 1 Fragment-size estimates for Acanthamoeba spp.


	Table 2 Mitochondrial genome sizes in Acanthamoeba spp. calculated by summation of five endonucleases


	Table 3 Proportions of homologous fragments. Estimates of interspecies genetic divergence in paired comparisons of ClaI, HaeIII, HindIII, PvuII and SalI digestion fragment patterns

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