Immunoproteomic analysis of the second-generation merozoite proteins of Eimeria tenella
Immunoproteomic analysis of the second-generation merozoite proteins of Eimeria tenella
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DOI：
10.1016/j.vetpar.2009.05.016
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发表时间：
2009-10-14
影响因子：
2.6
通讯作者：
Li, Xiangrui
中科院分区：
文献类型：
--
作者：
Liu, Liheng;Xu, Lixin;Li, Xiangrui
采用双向凝胶电泳（2-DE）和westernblot技术，对柔嫩艾美耳球虫（Eimeria tenella）人工感染鸡血清的第二代裂殖子全蛋白进行了分析。tenella。在考马斯亮蓝G-250染色的第二代裂殖子蛋白质组图谱上检测到约640个蛋白质点，在蛋白质印迹图谱上检测到85个蛋白质点。用基质辅助激光解吸电离飞行时间质谱（MALDI-TOF-MS）和MALDI-TOF-TOF-MS鉴定了44个抗原点。tenella和其他顶复门寄生虫或原生动物的3个同源蛋白。这些蛋白包括乳酸脱氢酶、烯醇化酶、14-3-3蛋白、微管蛋白β链、SERPIN 1蛋白前体、大亚基核糖体蛋白L23和E.柔嫩艾美耳球虫的热休克蛋白（HSP 70）、弓形虫的蛋白磷酸酶1型和草履虫的假定蛋白GSPATT 00020155001。其余蛋白质与E. tenella蛋白质序列数据库中使用“Mascot in-house”（2.1版）搜索引擎以自动模式检索，鉴定了11种未知蛋白质。在NCBI非冗余序列数据库（non-redundant sequence databases，NCBI）中对未知蛋白的氨基酸序列进行BLAST检索后，获得了大肠杆菌表面抗原12。tenella的膜骨架蛋白IMC 2A、线粒体F1-ATP合成酶β亚基前体、3-氧代酰基-酰基-载体蛋白合成酶和过氧化氢酶、恶性疟原虫3137的Vps 26和粘毛虫的假想蛋白TRIADDRAFT_60424。在44个蛋白质中，有8个蛋白质没有发现同源物，丰富了大肠杆菌第二代裂殖子免疫原性蛋白质的数据。tenella。(C)2009爱思唯尔有限公司版权所有。
Whole proteins of the second-generation merozoite of Eimeria tenella were analyzed by two-dimensional gel electrophoresis (2-DE) and western blot using the chicken sera infected artificially with E. tenella. Approximately 640 spots were detected on proteome map of the second-generation merozoite stained by Coomassie brilliant blue G-250 and 85 spots were recognized on western blot map as antigens. Forty four spots of the antigens were identified by matrix-assisted laser desorption ionization time-of-flight MS (MALDI-TOF-MS) and MALDI-TOF-TOF-MS. Twenty six proteins of E. tenella and three homologous proteins to other apicomplexan parasites or protozoan were identified using 'Mascot' server. These proteins included lactate dehydrogenase, enolase, 14-3-3 protein, microneme proteins, tubulin beta chain, SERPIN1 protein precursor, large subunit ribosomal protein L23 and surface antigens of E. tenella, heat shock protein (HSP70) of Eimeria acervulina, protein phosphatase type 1 of Toxoplasma gondii and hypothetical protein GSPATT00020155001 of Paramecium tetraurelia. The rest proteins were searched against the E. tenella protein sequence database using 'Mascot in-house' (version 2.1) search engine in automated mode and 11 unknown proteins were identified. After the amino acid sequence of the unknown proteins were searched using BLAST against non-redundant sequence databases (NCBI), surface antigen 12 of E. tenella and six homologous proteins to other apicomplexa parasites were found. They were membrane skeleton protein IMC2A, mitochondrial F1-ATP synthase beta subunit precursor, 3-oxoacyl-acyl-carrier-protein synthase and catalase of T gondii, Vps26 of Plasmodium falciparum 3137, and hypothetical protein TRIADDRAFT_60424 of Trichoplax adhaerens. No homologues of 8 spots of the 44 analyzed proteins were found. These proteins enriched the data of immunogenic proteins of the second-generation merozoite of E. tenella. (C) 2009 Elsevier B.V. All rights reserved.