Identification and characterization of Eimeria tenella apical membrane antigen-1 (AMA1)
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...
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Published in | PloS one Vol. 7; no. 7; p. e41115 |
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Main Authors | , , , , , , |
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19.07.2012
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Abstract | 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. |
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AbstractList | 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. 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. |
Audience | Academic |
Author | Han, Hongyu Zhu, Shunhai Lin, Jiaojiao Zhao, Qiping Dong, Hui Huang, Bing Jiang, Lianlian |
AuthorAffiliation | Key Laboratory for Animal Parasitology, Ministry of Agriculture, Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Minhang District, Shanghai, China University of Melbourne, Australia |
AuthorAffiliation_xml | – name: Key Laboratory for Animal Parasitology, Ministry of Agriculture, Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Minhang District, Shanghai, China – name: University of Melbourne, Australia |
Author_xml | – sequence: 1 givenname: Lianlian surname: Jiang fullname: Jiang, Lianlian organization: Key Laboratory for Animal Parasitology, Ministry of Agriculture, Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Minhang District, Shanghai, China – sequence: 2 givenname: Jiaojiao surname: Lin fullname: Lin, Jiaojiao – sequence: 3 givenname: Hongyu surname: Han fullname: Han, Hongyu – sequence: 4 givenname: Hui surname: Dong fullname: Dong, Hui – sequence: 5 givenname: Qiping surname: Zhao fullname: Zhao, Qiping – sequence: 6 givenname: Shunhai surname: Zhu fullname: Zhu, Shunhai – sequence: 7 givenname: Bing surname: Huang fullname: Huang, Bing |
BackLink | https://www.ncbi.nlm.nih.gov/pubmed/22829917$$D View this record in MEDLINE/PubMed |
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Notes | ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 Conceived and designed the experiments: JL BH LJ. Performed the experiments: LJ HH HD QZ SZ. Analyzed the data: LJ. Contributed reagents/materials/analysis tools: HH QZ. Wrote the paper: LJ. |
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SubjectTerms | 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 |
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Title | Identification and characterization of Eimeria tenella apical membrane antigen-1 (AMA1) |
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