Identification and characterization of Eimeria tenella apical membrane antigen-1 (AMA1)

• Published in: PloS one Vol. 7; no. 7; p. e41115
• Main Authors: Jiang, Lianlian, Lin, Jiaojiao, Han, Hongyu, Dong, Hui, Zhao, Qiping, Zhu, Shunhai, Huang, Bing
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 19.07.2012; Public Library of Science (PLoS)
• Subjects: Agriculture; Amino acids; Animals; Antibodies; Antigens; Antigens, Protozoan - genetics; Antigens, Protozoan - metabolism; Biology; Blotting, Western; Cell culture; Chickens; Developmental stages; E coli; Eimeria; Eimeria tenella; Eimeria tenella - genetics; Eimeria tenella - growth & development; Eimeria tenella - metabolism; Enzymes; Escherichia coli; Expressed sequence tags; Fluorescent Antibody Technique; Genomes; Immunofluorescence; Immunohistochemistry; Infections; Laboratories; Malaria; Merozoites; Oocysts; Parasites; Parasitology; Plasmodium falciparum; Polyclonal antibodies; Poultry; Protein expression; Proteins; Real-Time Polymerase Chain Reaction; Sporozoites; Sporozoites - growth & development; Sporozoites - metabolism; Toxoplasma gondii; Trends; Vaccines; Veterinary Science; Western blotting; China
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0041115

Apical membrane antigen-1 (AMA1) is a micronemal protein of apicomplexan parasites that appears to be essential during the invasion of host cells. In this study, a full-length cDNA of AMA1 was identified from Eimeria tenella (Et) using expressed sequence tag and the rapid amplification of cDNA ends technique. EtAMA1 had an open reading frame of 1608 bp encoding a protein of 535 amino acids. Quantitative real-time PCR analysis revealed that EtAMA1 was expressed at higher levels in sporozoites than in the other developmental stages (unsporulated oocysts, sporulated oocysts and second-generation merozoites). The ectodomain sequence was expressed as recombinant EtAMA1 (rEtAMA1) and rabbit polyclonal antibodies raised against the rEtAMA1 recognized a 58-kDa native parasite protein by Western Blotting and had a potent inhibitory effect on parasite invasion, decreasing it by approximately 70%. Immunofluorescence analysis and immunohistochemistry analysis showed EtAMA1 might play an important role in sporozoite invasion and development.