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Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1990 Jul;87(13):4976–4980. doi: 10.1073/pnas.87.13.4976

Cloning and expression of a membrane antigen of Entamoeba histolytica possessing multiple tandem repeats.

S L Stanley Jr 1, A Becker 1, C Kunz-Jenkins 1, L Foster 1, E Li 1
PMCID: PMC54244  PMID: 1695007

Abstract

Entamoeba histolytica causes amebic dysentery and amebic liver abscess, major causes of morbidity and mortality worldwide. We have used differential hybridization screening to isolate an E. histolytica-specific cDNA clone. The cDNA was found to encode a serine-rich E. histolytica protein (SREHP) containing multiple tandem repeats. The structural motif of SREHP resembles some of the repetitive antigens of malarial species, especially the circumsporozoite proteins. A recombinant trpE fusion protein containing the tandem repeats of SREHP was recognized by immune serum from a patient with amebiasis, demonstrating that SREHP is a naturally immunogenic protein. An antiserum raised against the recombinant fusion protein specifically bound to two distinct bands with apparent molecular masses of 46 and 52 kDa in a crude preparation of E. histolytica trophozoite membranes. This antiserum also inhibited E. histolytica trophozoite adhesion to Chinese hamster ovary cells in vitro. The ability to isolate E. histolytica-specific genes, and to express those genes in Escherichia coli, may be important in studying the molecular basis of E. histolytica pathogenesis and for the future development of vaccines.

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Selected References

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  1. 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]
  2. 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]
  3. Diamond L. S. Techniques of axenic cultivation of Entamoeba histolytica Schaudinn, 1903 and E. histolytica-like amebae. J Parasitol. 1968 Oct;54(5):1047–1056. [PubMed] [Google Scholar]
  4. Feinberg A. P., Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. doi: 10.1016/0003-2697(83)90418-9. [DOI] [PubMed] [Google Scholar]
  5. Galinski M. R., Arnot D. E., Cochrane A. H., Barnwell J. W., Nussenzweig R. S., Enea V. The circumsporozoite gene of the Plasmodium cynomolgi complex. Cell. 1987 Jan 30;48(2):311–319. doi: 10.1016/0092-8674(87)90434-x. [DOI] [PubMed] [Google Scholar]
  6. Ghadirian E., Meerovitch E., Hartmann D. P. Protection against amebic liver abscess in hamsters by means of immunization with amebic antigen and some of its fractions. Am J Trop Med Hyg. 1980 Sep;29(5):779–784. doi: 10.4269/ajtmh.1980.29.779. [DOI] [PubMed] [Google Scholar]
  7. 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]
  8. Hanahan D. Studies on transformation of Escherichia coli with plasmids. J Mol Biol. 1983 Jun 5;166(4):557–580. doi: 10.1016/s0022-2836(83)80284-8. [DOI] [PubMed] [Google Scholar]
  9. Hardy W. R., Strauss J. H. Processing the nonstructural polyproteins of Sindbis virus: study of the kinetics in vivo by using monospecific antibodies. J Virol. 1988 Mar;62(3):998–1007. doi: 10.1128/jvi.62.3.998-1007.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Heuckeroth R. O., Birkenmeier E. H., Levin M. S., Gordon J. I. Analysis of the tissue-specific expression, developmental regulation, and linkage relationships of a rodent gene encoding heart fatty acid binding protein. J Biol Chem. 1987 Jul 15;262(20):9709–9717. [PubMed] [Google Scholar]
  11. Kemp D. J., Coppel R. L., Anders R. F. Repetitive proteins and genes of malaria. Annu Rev Microbiol. 1987;41:181–208. doi: 10.1146/annurev.mi.41.100187.001145. [DOI] [PubMed] [Google Scholar]
  12. Krupp I. M. Protective immunity to amebic infection demonstrated in guinea pigs. Am J Trop Med Hyg. 1974 May;23(3):355–360. doi: 10.4269/ajtmh.1974.23.355. [DOI] [PubMed] [Google Scholar]
  13. Li E., Becker A., Stanley S. L., Jr Chinese hamster ovary cells deficient in N-acetylglucosaminyltransferase I activity are resistant to Entamoeba histolytica-mediated cytotoxicity. Infect Immun. 1989 Jan;57(1):8–12. doi: 10.1128/iai.57.1.8-12.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Li E., Becker A., Stanley S. L., Jr Use of Chinese hamster ovary cells with altered glycosylation patterns to define the carbohydrate specificity of Entamoeba histolytica adhesion. J Exp Med. 1988 May 1;167(5):1725–1730. doi: 10.1084/jem.167.5.1725. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Li E., Demmer L. A., Sweetser D. A., Ong D. E., Gordon J. I. Rat cellular retinol-binding protein II: use of a cloned cDNA to define its primary structure, tissue-specific expression, and developmental regulation. Proc Natl Acad Sci U S A. 1986 Aug;83(16):5779–5783. doi: 10.1073/pnas.83.16.5779. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Mattern C. F., Keister D. B., Caspar P. A. Experimental amebiasis. III. A rapid in vitro assay for virulence of Entamoeba histolytica. Am J Trop Med Hyg. 1978 Sep;27(5):882–887. [PubMed] [Google Scholar]
  17. Maxam A. M., Gilbert W. Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol. 1980;65(1):499–560. doi: 10.1016/s0076-6879(80)65059-9. [DOI] [PubMed] [Google Scholar]
  18. Ozaki L. S., Svec P., Nussenzweig R. S., Nussenzweig V., Godson G. N. Structure of the plasmodium knowlesi gene coding for the circumsporozoite protein. Cell. 1983 Oct;34(3):815–822. doi: 10.1016/0092-8674(83)90538-x. [DOI] [PubMed] [Google Scholar]
  19. Rodríguez M. A., Hernández F., Santos L., Valdez A., Orozco E. Entamoeba histolytica: molecules involved in the target cell-parasite relationship. Mol Biochem Parasitol. 1989 Nov;37(1):87–99. doi: 10.1016/0166-6851(89)90105-9. [DOI] [PubMed] [Google Scholar]
  20. SWARTZWELDER J. C., AVANT W. H. Immunity to amebic infection in dogs. Am J Trop Med Hyg. 1952 Jul;1(4):567–575. doi: 10.4269/ajtmh.1952.1.567. [DOI] [PubMed] [Google Scholar]
  21. Tannich E., Horstmann R. D., Knobloch J., Arnold H. H. Genomic DNA differences between pathogenic and nonpathogenic Entamoeba histolytica. Proc Natl Acad Sci U S A. 1989 Jul;86(13):5118–5122. doi: 10.1073/pnas.86.13.5118. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Thomas P. S. Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose. Proc Natl Acad Sci U S A. 1980 Sep;77(9):5201–5205. doi: 10.1073/pnas.77.9.5201. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Villa-Komaroff L., Efstratiadis A., Broome S., Lomedico P., Tizard R., Naber S. P., Chick W. L., Gilbert W. A bacterial clone synthesizing proinsulin. Proc Natl Acad Sci U S A. 1978 Aug;75(8):3727–3731. doi: 10.1073/pnas.75.8.3727. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. von Heijne G. A new method for predicting signal sequence cleavage sites. Nucleic Acids Res. 1986 Jun 11;14(11):4683–4690. doi: 10.1093/nar/14.11.4683. [DOI] [PMC free article] [PubMed] [Google Scholar]

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