Cloning of the entire COWP gene of Cryptosporidium parvum and ultrastructural localization of the protein during sexual parasite development

• Published in: Parasitology Vol. 114 ( Pt 5); p. 427
• Main Authors: Spano, F, Puri, C, Ranucci, L, Putignani, L, Crisanti, A
• Format: Journal Article
• Language: English
• Published: England 01.05.1997
• Subjects: Amino Acid Sequence; Animals; Base Sequence; Cell Wall - ultrastructure; Cloning, Molecular; Cryptosporidium parvum - genetics; Cryptosporidium parvum - growth & development; Cryptosporidium parvum - ultrastructure; DNA, Protozoan - analysis; Epithelium - parasitology; Genes, Protozoan - genetics; Ileum - parasitology; Mice; Microscopy, Immunoelectron; Molecular Sequence Data; Protozoan Proteins - analysis; Protozoan Proteins - chemistry; Protozoan Proteins - genetics; Recombinant Fusion Proteins; Restriction Mapping; Sequence Analysis, DNA; Sequence Homology, Amino Acid
• ISSN: 0031-1820
• DOI: 10.1017/S0031182096008761

Molecular cloning and immunoelectron microscopy have been used to clone the full-length gene encoding Cryptosporidium parvum oocyst wall protein (COWP) and to analyse at the ultrastructural level the expression and localization of COWP during development in the gut. COWP is 1622 amino acids long, has a typical leader peptide and consists of 2 amino acidic domains each containing distinct repeated elements possibly originating from a common ancestral precursor. Electron microscopy localized COWP in a large cytoplasmic inclusion and in the wall-forming bodies of early and late macrogametes, respectively. Ultrastructural analysis of double-walled sporulating and mature oocysts indicated that COWP is selectively localized in the inner layer of the oocyst wall. This study provides the first localization at the ultrastructural level of a cloned coccidian oocyst wall protein.