Molecular cloning of the gene for a cryptosporidium sporozoite surface protein.

Important note: Information in this article was accurate in 1991. The state of the art may have changed since the publication date.

Molecular cloning of the gene for a cryptosporidium sporozoite surface protein.

Int Conf AIDS. 1991 Jun 16-21;7(1):142 (abstract no. M.A.1200). Unique Identifier : AIDSLINE ICA7/1120091
Petersen C; Gut J; Leech JH; Nelson RG; Parasitology Laboratory, San Francisco General Hospital and; Departments of Medicine and Pharmaceutical Chemistry, University; of California, San Francisco, CA, USA

Abstract: OBJECTIVE: To clone the gene for a surface glycoprotein of Cryptosporidium sporozoites that is the target of a monoclonal antibody (Mab 10C6) that reacts with intracellular merozoites and the surface of sporozoites, causes shedding of the sporozoite surface coat and inhibits sporozoite invasion of Mardin-Darby Canine Kidney (MDCK) cells in vitro. METHODS: A lambda gt 11 gDNA library was screened with polyclonal mouse antisera raised to sporozoites and oocysts. Positive clones were plaque purified and used to prepare recombinant eluted antibody (REA) from the polyclonal antisera. RESULTS: REA from four clones reacted with a greater than 300 kD protein on Western blot and exhibited an IFA pattern identical to that of Mab 10C6 on fixed sporozoites. REA from all four clones reacted on Western blot with the greater than 300 kD protein immunoprecipitated by Mab 10C6. Following N-glycosidase F treatment, REA from the clones recognized multiple Cryptosporidium proteins of Mr 150-190, but Mab 10C6 did not react with any. CONCLUSIONS: The gene for a potentially protective antigen on the surface of Cryptosporidium sporozoites has been cloned and encodes a heavily glycosylated protein. The Mab 10C6 epitope requires N-linked glycosylation.

Keywords: Animal Antibodies, Monoclonal/IMMUNOLOGY Antigens, Protozoan/*GENETICS Antigens, Surface/*GENETICS Blotting, Western Cells, Cultured Cloning, Molecular Cryptosporidium/*GENETICS/IMMUNOLOGY Dogs Epitopes Glycoside Hydrolases/IMMUNOLOGY Glycosylation Membrane Glycoproteins/*GENETICS Mice Molecular Weight Precipitin Tests Protozoan Proteins/*GENETICS/IMMUNOLOGY Recombinant Proteins/IMMUNOLOGY ABSTRACT
912130
M91C3755

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