基于噬菌体展示技术的环形泰勒虫配基和蜱源受体的鉴定

项目名称： 基于噬菌体展示技术的环形泰勒虫配基和蜱源受体的鉴定

项目编号： No.31201899

项目类型： 青年科学基金项目

立项/批准年度： 2013

项目学科： 畜牧学与草地科学、兽医学、水产学

项目作者： 田占成

作者单位： 中国农业科学院兰州兽医研究所

项目金额： 24万元

中文摘要： 梨形虫病是由梨形虫纲巴贝斯科或泰勒科原虫引起的蜱传性血液原虫病的总称，给畜牧业的发展造成极大的经济损失。但有关梨形虫对媒介蜱组织器官的侵入过程和感染机制的研究较少。本项目将噬菌体展示肽技术应用于环形泰勒虫配基的研究，从环形泰勒虫在小亚璃眼蜱体内发育成熟时期的蜱源细胞受体这一关键环节入手，通过多轮在小亚璃眼蜱体内的亲和筛选，富集与蜱源上皮细胞具有特异结合活性的配基，作为环形泰勒虫与表面受体结合的有效竞争物。应用器官ELISA、组织定位验证配基的靶向性，通过配基注射饥饿雌蜱的方法进行环形泰勒虫-蜱源受体阻断实验，以评估获得的配基活性分子抑制小亚璃眼蜱传播环形泰勒虫的作用。进一步，以鉴定的环形泰勒虫配基为钓饵，采用pull-down和质谱技术从小亚璃眼蜱唾液腺和中肠组织中获得和鉴定环形泰勒虫蜱源受体。最终为揭示环形泰勒虫入侵媒介蜱唾液腺和中肠的机理，靶向治疗和预防提供理论基础。

中文关键词： 残缘璃眼蜱；环形泰勒虫；传播；鉴定；抗原

英文摘要： Transmission of Piroplasm, the causative agent of Piroplasmosis, requires the completion of a complex life cycle in the tick, which includes invasion of the salivary glands and midguts. This invasion depends on the recognition of tick salivary gland and midgut surface components by the parasite. However, there is limited information on the invasion process and infection mechanism of Piroplasm in tick. By use of a phage display library, the work targeting Theileria annulata (T. annulata) ligands on Hyalomma anatolicum anatolicum (H. a. anatolicum), through 6 rounds of phage selection in vivo, and enriches the active ligands that specifically binds to the epithelia of tick, which binds to and competes for T. annulata receptor on the surface of the salivary glands and midguts. Binding Specificity of the ligands were verified by the method of Organ ELISA and Histochemical Staining, and it is further evaluated the roles that the ligands inhibit T. annulata Sporozoite invasion of salivary glands through the ligands were injected into the unfed female ticks. Furthermore, the identified T. annulata ligands as bait, By use of UV-crosslinking experiments between the peptide and its salivary gland and midgut target protein, to identify T. annulata receptors on the epithelia of the midgut and salivary glands of H. a. anatol

英文关键词： Hyalomma detritum；Theileria annulata；transmission；identification；antigen