Cloning and expression of a membrane antigen of Entamoeba histolytica possessing multiple tandem repeats.
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.
Department of Medicine, Washington University School of Medicine, Saint Louis, MO 63110.
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Stanley
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Becker
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Kunz-Jenkins
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Foster
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Li
D000595:Amino Acid Sequence; D000818:Animals; D000953:Antigens, Protozoan / Q000235:genetics* / Q000302:isolation & purification; D000954:Antigens, Surface / Q000235:genetics* / Q000302:isolation & purification; D001483:Base Sequence; D015152:Blotting, Northern; D015153:Blotting, Western; D002448:Cell Adhesion; D002460:Cell Line; D003001:Cloning, Molecular / Q000379:methods; D004247:DNA / Q000235:genetics; D004748:Entamoeba histolytica / Q000235:genetics*; D015870:Gene Expression; D015723:Gene Library; D008565:Membrane Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D012313:RNA / Q000235:genetics / Q000302:isolation & purification; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012091:Repetitive Sequences, Nucleic Acid
D000953:Antigens, Protozoan; D000954:Antigens, Surface; D008565:Membrane Proteins; D015345:Oligonucleotide Probes; D011993:Recombinant Fusion Proteins; D012313:RNA; D004247:DNA