Hostname: page-component-78c5997874-s2hrs Total loading time: 0 Render date: 2024-11-05T03:48:26.377Z Has data issue: false hasContentIssue false
  • English
  • Français

Isolation and characterization of a species-specific multicopy DNA sequence from Entamoeba histolytica

Published online by Cambridge University Press:  06 April 2009

V. Mittal ,
A. Bhattacharya  and
S. Bhattacharya
V. Mittal
Affiliation:
School of Life Sciences, Jawaharlal Nehru University, New Delhi 110067, India
A. Bhattacharya
Affiliation:
School of Life Sciences, Jawaharlal Nehru University, New Delhi 110067, India
S. Bhattacharya
Affiliation:
School of Environmental Sciences, Jawaharlal Nehru University, New Delhi 110067, India
Get access Rights & Permissions [Opens in a new window]

Summary

A genomic library of Entamoeba histolytica (pathogenic strain HM-1: IMSS) was screened to detect repetitive DNA clones other than those from the highly abundant ribosomal DNA (rDNA). One such clone (HMc) had a 2·3 kb insert which hybridized with the main genome and not the rDNA circle. Southern hybridization of E. histolytica genomic DNA, digested with EcoR I and probed with HMc, showed multiple bands. The banding pattern was identical in all axenic pathogenic strains tested. Differences, however, existed when the banding pattern of a pathogenic strain was compared with that of a non-pathogenic strain. HMc was present in about 25–30 copies per genome in strain HM-1: IMSS. Nucleotide sequence analysis of HMc revealed a partial open reading frame which hybridized with a 1·35 kb poly A+ transcript in Northern blots. The deduced amino acid sequence did not, however, show significant homology with known proteins. The HMc sequence was found only in E. histolytica as it hybridized with 5 different axenic strains of E. histolytica but did not recognize other closely related species of Entamoeba. It has thus the potential to be used as a species-specific DNA probe.

Keywords

Entamoeba histolyticarepetitive DNAgenomic organizationspecies-specific sequence
Type
Research Article
Information
Parasitology , Volume 108 , Issue 3 , April 1994 , pp. 237 - 244
Copyright
Copyright © Cambridge University Press 1994

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

REFERENCES

Bhattacharya, S., Bhattacharya, A. & Diamond, L. S. (1988). Comparison of repeated DNA from strains of Entamoeba histolytica and other Entamoeba. Molecular and Biochemical Parasitology 27, 257–62.CrossRefGoogle ScholarPubMed
Bhattacharya, S., Bhattacharya, A., Diamond, L. S. & Soldo, A. T. (1989). Circular DNA of Entamoeba histolytica encodes ribosomal RNA. Journal of Protozoology 36, 455–8.CrossRefGoogle ScholarPubMed
Chomczynski, P. & Sacchi, N. (1987). Single step method of RNA isolation by guanidinium thiocyanate-phenol-chloroform extraction. Analytical Biochemistry 162, 156–9.CrossRefGoogle ScholarPubMed
Clark, C. G. & Diamond, L. S. (1991). Ribosomal RNA genes of ‘pathogenic’ and ‘nonpathogenic’ Entamoeba histolytica are distinct. Molecular and Biochemical Parasitology 49, 297–302.CrossRefGoogle ScholarPubMed
Cruz-Reyes, J. A., Spice, W. M., Rehman, T., Gisborne, E. & Ackers, J. P. (1992). Ribosomal DNA sequences in the differentiation of pathogenic and nonpathogenic isolates of Entamoeba histolytica. Parasitology 104, 239–46.CrossRefGoogle ScholarPubMed
Descoteaux, S., Ayala, P., Orozco, E. & Samuelson, J. (1992). Primary sequence of 2 P-glycoprotein genes of Entamoeba histolytica. Molecular and Biochemical Parasitology 54, 201–12.CrossRefGoogle Scholar
Diamond, L. S. (1982). A new liquid medium for xenic cultivation of Entamoeba histolytica and other lumen dwelling protozoa. Journal of Parasitology 68, 958–9.Google Scholar
Diamond, L. S. (1983). Lumen dwelling protozoa: Entamoeba, Trichomonads and Giardia. In In Vitro Cultivation of Protozoan Parasites (ed. Jensen, J. B.), pp. 65–109. Boca Raton, Florida: CRC press.Google Scholar
Diamond, L. S. & Clark, C. G. (1993). A redescription of Entamoeba histolytica Schaudinn, 1903 (Emended Walker, 1911) separating it from Entamoeba dispar Brumpt, 1925. Journal of Eukaryotic Microbiology 40, 340–4.CrossRefGoogle ScholarPubMed
Edman, U., Meza, I. & Agabian, A. (1987). Genomic and cDNA actin sequences from a virulent strain of Entamoeba histolytica. Proceedings of the National Academy of Sciences, USA 84, 3024–8.Google Scholar
Edman, U., Meraz, A. M., Rausser, S., Agabian, A. & Meza, I. (1990). Characterization of an immuno-dominant variable surface antigen from pathogenic and nonpathogenic Entamoeba histolytica. Journal of Experimental Medicine 172, 879–88.Google Scholar
Garfinkel, L. I., Giladi, M., Huber, M., Gitler, C., Mirelman, D., Revel, M. & Rozenblatt, S. (1989). DNA probes specific for Entamoeba histolytica possessing pathogenic and nonpathogenic zymodemes. Infection and Immunity 57, 926–31.CrossRefGoogle ScholarPubMed
Gelderman, A. H., Bartgis, I. L., Keister, D. B. & Diamond, L. S. (1971). A comparison of genome sizes and thermal-denaturation-derived base composition of DNAs from several members of Entamoeba (histolytica group). Journal of Parasitology 57, 912–16.CrossRefGoogle ScholarPubMed
Hanahan, D. (1985). Techniques for transformation of E. coli. In DNA Cloning. A Practical Approach, Vol 1 (ed. Glover, D. M.), pp. 109–135. Oxford: IRL Press.Google Scholar
Henikoff, S. (1984). Unidirectional digestion with Exonuclease III creates targeted breakpoints for DNA sequencing. Gene 28, 351–9.Google Scholar
Huber, M., Garfinkel, L., Gitler, C., Mirelman, D., Revel, M. & Rozenblatt, S. (1987). Entamoeba histolytica: cloning and characterisation of actin cDNA. Molecular and Biochemical Parasitology 24, 227–35.Google Scholar
Huber, M., Garfinkel, L., Gitler, C., Mirelman, D., Revel, M. & Rozenblatt, M. (1988). Nucleotide sequence of Entamoeba histolytica ferredoxin gene. Molecular and Biochemical Parasitology 31, 27–34.CrossRefGoogle ScholarPubMed
Huber, J., Koller, B., Gitler, C., Mirelman, D., Revel, M., Rozenblatt, S. & Garfinkel, L. (1989). E. histolytica ribosomal RNA genes are carried on palindromic circular DNA molecules. Molecular and Biochemical Parasitology 32, 285–96.Google Scholar
Maniatis, T., Fritsch, E. F. & Sambrook, J. (1982). Molecular Cloning: A Laboratory Manual. Cold Spring Harbour, New York: Cold Spring Harbour Laboratory.Google Scholar
Michel, B., Alagon, A., Lizardi, P. M. & Zurita, M. (1992). Characterisation of a repetitive DNA element from Entamoeba histolytica. Molecular and Biochemical Parasitology 51, 165–8.Google Scholar
Mittal, V., Ramachandran, S., Sehgal, D., Bhattacharya, A. & Bhattacharya, S. (1991). Sequence analysis of a DNA fragment with yeast autonomously replicating sequence activity from the extrachromosomal ribosomal DNA circle of Entamoeba histolytica. Nucleic Acids Research 19, 2777.Google Scholar
Mittal, V., Sehgal, D., Bhattacharya, A. & Bhattacharya, S. (1992). A second short repeat detected downstream of rRNA genes in the Entamoeba histolytica rDNA circle. Molecular and Biochemical Parasitology 54, 97–100.CrossRefGoogle Scholar
Prasad, J., Bhattacharya, A. & Bhattacharya, S. (1992). Cloning and sequence analysis of a calcium binding protein gene from a pathogenic strain of E. histolytica. Molecular and Biochemical Parasitology 52, 137–40.CrossRefGoogle Scholar
Sanger, F., Nicklen, D. & Coulson, A. R. (1977). DNA sequencing with chain terminating inhibitors. Proceedings of the National Academy of Sciences, USA 74, 5463–7.Google Scholar
Sargeaunt, P. G. (1987). The reliability of Entamoeba histolytica zymodemes in clinical diagnosis. Parasitology Today 3, 40–3.Google Scholar
Stanley, S. L. Jr, Becker, A., Kunz-Jenkins, C., Foster, L. & Li, E. (1990). Cloning and expression of a membrane antigen of Entamoeba histolytica possessing multiple tandem repeats. Proceedings of the National Academy of Sciences, USA 87, 4976–80.CrossRefGoogle ScholarPubMed
Strachan, D., Spice, W. M., Chiodini, P. L., Moody, A. H. & Ackers, J. P. (1988). Immunological differentiation of pathogenic and nonpathogenic isolates of Entamoeba histolytica. Lancet 1, 561–2.Google Scholar
Tannich, E., Horstmann, R. D., Knobloch, J. & Arnold, H. H. (1989). Genomic DNA differences between pathogenic and nonpathogenic Entamoeba histolytica. Proceedings of the National Academy of Sciences, USA 86, 5118–22.CrossRefGoogle ScholarPubMed
Tannich, E., Ebert, F. & Horstmann, R. D. (1991 a). Primary structure of the 170-kda surface lectin of pathogenic Entamoeba histolytica. Proceedings of the National Academy of Sciences, USA 88, 1849–53.Google Scholar
Tannich, E., Scholze, H., Nickel, R. & Horstmann, R. D. (1991 b). Homologous cysteine proteinases of pathogenic and nonpathogenic Entamoeba histolytica. Differences in structure and expression. Journal of Biological Chemistry 266, 4798–803.Google Scholar
Tannich, E., Bruchhaus, I., Walter, R. D. & Horstman, R. D. (1991 c). Pathogenic and nonpathogenic Entamoeba histolytica: identification and molecular cloning of an iron-containing superoxide dismutase. Molecular and Biochemical Parasitology 49, 61–71.Google Scholar
Walsh, J. A. (1986). Problems in recognition and diagnosis of amebiasis: estimation of the global magnitude of morbidity and mortality. Reviews of Infectious Diseases 8, 228–38.Google Scholar