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. 1989 Jul;86(13):5118–5122. doi: 10.1073/pnas.86.13.5118

Genomic DNA differences between pathogenic and nonpathogenic Entamoeba histolytica.

E Tannich 1, R D Horstmann 1, J Knobloch 1, H H Arnold 1
PMCID: PMC297568  PMID: 2544890

Abstract

cDNA libraries were constructed from pathogenic (HM-1:IMSS) and nonpathogenic (SAW 1734) isolates of Entamoeba histolytica. A cDNA clone (cEH-P1) specific for pathogenic amoebae was identified by screening with a pool of sera from patients with invasive amoebiasis that had been absorbed with nonpathogenic amoebae. This clone was used for the identification of a homologous clone (cEH-NP1) in the cDNA from nonpathogenic amoebae. Sequence analysis and comparison of the predicted amino acid sequences for both clones disclosed 12% evolutionary divergence in structure. Hybridization of both cDNA probes to genomic DNA from four pathogenic and five nonpathogenic E. histolytica isolates revealed two distinct Southern blot patterns, one characteristic for pathogenic amoebae and the other for nonpathogenic amoebae. Further, the complex pattern of restriction fragments hybridizing to an actin cDNA probe was also different between pathogenic and nonpathogenic isolates but was conserved within each group of amoebae. The results indicate that pathogenic isolates of E. histolytica are genetically distinct from nonpathogenic isolates.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Allason-Jones E., Mindel A., Sargeaunt P., Williams P. Entamoeba histolytica as a commensal intestinal parasite in homosexual men. N Engl J Med. 1986 Aug 7;315(6):353–356. doi: 10.1056/NEJM198608073150603. [DOI] [PubMed] [Google Scholar]
  2. Bhattacharya S., Bhattacharya A., Diamond L. S. Comparison of repeated DNA from strains of Entamoeba histolytica and other Entamoeba. Mol Biochem Parasitol. 1988 Jan 15;27(2-3):257–262. doi: 10.1016/0166-6851(88)90045-x. [DOI] [PubMed] [Google Scholar]
  3. Chirgwin J. M., Przybyla A. E., MacDonald R. J., Rutter W. J. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979 Nov 27;18(24):5294–5299. doi: 10.1021/bi00591a005. [DOI] [PubMed] [Google Scholar]
  4. Denhardt D. T. A membrane-filter technique for the detection of complementary DNA. Biochem Biophys Res Commun. 1966 Jun 13;23(5):641–646. doi: 10.1016/0006-291x(66)90447-5. [DOI] [PubMed] [Google Scholar]
  5. Diamond L. S. A new liquid medium for xenic cultivation of Entamoeba histolytica and other lumen-dwelling protozoa. J Parasitol. 1982 Oct;68(5):958–959. [PubMed] [Google Scholar]
  6. Diamond L. S., Harlow D. R., Cunnick C. C. A new medium for the axenic cultivation of Entamoeba histolytica and other Entamoeba. Trans R Soc Trop Med Hyg. 1978;72(4):431–432. doi: 10.1016/0035-9203(78)90144-x. [DOI] [PubMed] [Google Scholar]
  7. Edman U., Meza I., Agabian N. Genomic and cDNA actin sequences from a virulent strain of Entamoeba histolytica. Proc Natl Acad Sci U S A. 1987 May;84(9):3024–3028. doi: 10.1073/pnas.84.9.3024. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Erttmann K. D., Unnasch T. R., Greene B. M., Albiez E. J., Boateng J., Denke A. M., Ferraroni J. J., Karam M., Schulz-Key H., Williams P. N. A DNA sequence specific for forest form Onchocerca volvulus. Nature. 1987 Jun 4;327(6121):415–417. doi: 10.1038/327415a0. [DOI] [PubMed] [Google Scholar]
  9. Garfinkel L. I., Giladi M., Huber M., Gitler C., Mirelman D., Revel M., Rozenblatt S. DNA probes specific for Entamoeba histolytica possessing pathogenic and nonpathogenic zymodemes. Infect Immun. 1989 Mar;57(3):926–931. doi: 10.1128/iai.57.3.926-931.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Gelderman A. H., Keister D. B., Bartgis I. L., Diamond L. S. Characterization of the deoxyribonucleic acid of representative strains of Entamoeba histolytica, E. histoytica-like amebae, and E. moshkovskii. J Parasitol. 1971 Aug;57(4):906–911. [PubMed] [Google Scholar]
  11. Gubler U., Hoffman B. J. A simple and very efficient method for generating cDNA libraries. Gene. 1983 Nov;25(2-3):263–269. doi: 10.1016/0378-1119(83)90230-5. [DOI] [PubMed] [Google Scholar]
  12. Huber M., Garfinkel L., Gitler C., Mirelman D., Revel M., Rozenblatt S. Entamoeba histolytica: cloning and characterization of actin cDNA. Mol Biochem Parasitol. 1987 Jul;24(3):227–235. doi: 10.1016/0166-6851(87)90154-x. [DOI] [PubMed] [Google Scholar]
  13. Macina R. A., Arauzo S., Reyes M. B., Sanchez D. O., Basombrio M. A., Montamat E. E., Solari A., Frasch A. C. Trypanosoma cruzi isolates from Argentina and Chile grouped with the aid of DNA probes. Mol Biochem Parasitol. 1987 Aug;25(1):45–53. doi: 10.1016/0166-6851(87)90017-x. [DOI] [PubMed] [Google Scholar]
  14. Messing J. New M13 vectors for cloning. Methods Enzymol. 1983;101:20–78. doi: 10.1016/0076-6879(83)01005-8. [DOI] [PubMed] [Google Scholar]
  15. Mirelman D., Bracha R., Chayen A., Aust-Kettis A., Diamond L. S. Entamoeba histolytica: effect of growth conditions and bacterial associates on isoenzyme patterns and virulence. Exp Parasitol. 1986 Aug;62(1):142–148. doi: 10.1016/0014-4894(86)90017-2. [DOI] [PubMed] [Google Scholar]
  16. Mirelman D., Bracha R., Wexler A., Chayen A. Changes in isoenzyme patterns of a cloned culture of nonpathogenic Entamoeba histolytica during axenization. Infect Immun. 1986 Dec;54(3):827–832. doi: 10.1128/iai.54.3.827-832.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Mirelman D. Effect of culture conditions and bacterial associates on the zymodemes of Entamoeba histolytica. Parasitol Today. 1987 Feb;3(2):37–37. doi: 10.1016/0169-4758(87)90210-9. [DOI] [PubMed] [Google Scholar]
  18. Moss D. M., Mathews H. M. A fast electrophoretic isoenzyme technique for the identification of invasive and non-invasive Entamoeba histolytica and "E. histolytica-like" organisms. J Protozool. 1987 Aug;34(3):253–255. doi: 10.1111/j.1550-7408.1987.tb03169.x. [DOI] [PubMed] [Google Scholar]
  19. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Sargeaunt P. G., Jackson T. F., Simjee A. Biochemical homogeneity of Entamoeba histolytica isolates, especially those from liver abscess. Lancet. 1982 Jun 19;1(8286):1386–1388. doi: 10.1016/s0140-6736(82)92502-8. [DOI] [PubMed] [Google Scholar]
  21. Sargeaunt P. G. The reliability of Entamoeba histolytica zymodemes in clinical diagnosis. Parasitol Today. 1987 Feb;3(2):40–37. doi: 10.1016/0169-4758(87)90211-0. [DOI] [PubMed] [Google Scholar]
  22. Sargeaunt P. G., Williams J. E., Grene J. D. The differentiation of invasive and non-invasive Entamoeba histolytica by isoenzyme electrophoresis. Trans R Soc Trop Med Hyg. 1978;72(5):519–521. doi: 10.1016/0035-9203(78)90174-8. [DOI] [PubMed] [Google Scholar]
  23. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  24. Strachan W. D., Chiodini P. L., Spice W. M., Moody A. H., Ackers J. P. Immunological differentiation of pathogenic and non-pathogenic isolates of Entamoeba histolytica. Lancet. 1988 Mar 12;1(8585):561–563. doi: 10.1016/s0140-6736(88)91355-4. [DOI] [PubMed] [Google Scholar]
  25. Walsh J. A. Problems in recognition and diagnosis of amebiasis: estimation of the global magnitude of morbidity and mortality. Rev Infect Dis. 1986 Mar-Apr;8(2):228–238. doi: 10.1093/clinids/8.2.228. [DOI] [PubMed] [Google Scholar]
  26. Wirth D. F., Rogers W. O., Barker R., Jr, Dourado H., Suesebang L., Albuquerque B. Leishmaniasis and malaria: new tools for epidemiologic analysis. Science. 1986 Nov 21;234(4779):975–979. doi: 10.1126/science.3535070. [DOI] [PubMed] [Google Scholar]
  27. Young R. A., Davis R. W. Efficient isolation of genes by using antibody probes. Proc Natl Acad Sci U S A. 1983 Mar;80(5):1194–1198. doi: 10.1073/pnas.80.5.1194. [DOI] [PMC free article] [PubMed] [Google Scholar]

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