Cloning and characterization of Clonorchis sinensis myoglobin using immune sera against excretory-secretory antigens.
Clonorchis sinensis excretory-secretory (ES) antigens were separated by gradient SDS-PAGE and the antigenic protein isolated at 21 kDa was injected into rats. A C. sinensis cDNA library was then immunoscreened with sera from the 21-kDa antigenic protein-immunized rats. The selected genes, which were named C. sinensis myoglobin, contained a single open reading frame of 450 base pairs encoding 150 amino acids. A single hybridized band of 0.57 kb and two to four hybridized bands were detected by Northern and Southern blotting. Purified recombinant C. sinensis myoglobin was recognized by clonorchiasic rabbit sera (50%) and clonorchiasic human sera (25%). An indirect immunofluorescence assay showed that C. sinensis myoglobin is distributed over the whole body of the adult worm. It is believed that the abundance of C. sinensis myoglobin plays an important role as an oxygen reservoir under anaerobic conditions.
Sim
Seobo
S
Department of Parasitology and Institute of Tropical Medicine, BK21 Project for Medical Science, Yonsei University College of Medicine, 120-752, Seoul, Korea.
Park
Gab-Man
GM
Yong
Tai-Soon
TS
D000818:Animals; D000947:Antigens, Helminth / Q000276:immunology / Q000302:isolation & purification; D015152:Blotting, Northern; D015139:Blotting, Southern; D003001:Cloning, Molecular; D003004:Clonorchis sinensis / Q000276:immunology* / Q000378:metabolism; D019084:Fluorescent Antibody Technique, Indirect; D015723:Gene Library; D015801:Helminth Proteins / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification; D006801:Humans; D009211:Myoglobin / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification; D011817:Rabbits; D051381:Rats; D017207:Rats, Sprague-Dawley; D011994:Recombinant Proteins / Q000032:analysis / Q000276:immunology / Q000378:metabolism
D000947:Antigens, Helminth; D015801:Helminth Proteins; D009211:Myoglobin; D011994:Recombinant Proteins