摘要
为了研究柔嫩艾美耳球虫顶膜抗原2(EtAMA2)的分子特性与功能,利用间接免疫荧光试验、入侵抑制试验、实时荧光定量PCR、Western blot以及分泌试验等方法分析其生物学特性。结果发现EtAMA2蛋白具一个跨膜区和信号肽,在子孢子和第二代裂殖子中的转录水平明显高于未孢子化卵囊和孢子化卵囊,但蛋白仅在第二代裂殖子中被检测到;EtAMA2在游离的子孢子和第二代裂殖子中主要分布在胞质,在胞内的子孢子中主要分布在顶端。EtAMA2是一种微线体分泌蛋白,其分泌受到温度的调控。抗EtAMA2特异性抗体对子孢子入侵细胞没有明显抑制作用;将EtAMA2转染至DF-1细胞后,子孢子入侵率明显提高。本研究结果为深入研究 EtAMA2 在球虫入侵过程中的功能与作用机制奠定基础。
In order to study the molecular characteristics and biological functions of apical membrane antigen 2 of Eimeria tenella(Et AMA2),indirect immunofl uorescence assay,invasion inhibition assay,quantitative real-time PCR,Western blot and secretion assays were used to study its biological properties.The results indicated that Et AMA2 protein had a transmembrane region and a signal peptide,and its transcription level was higher in sporozoites and second-generation merozoites than in unsporulated oocysts and sporulated oocysts,but its protein was expressed only in second-generation merozoites.Et AMA2 was mainly distributed on the cytoplasm of free sporozoites and second-generation merozoites at the apical end of intracellular sporozoites.Et AMA2 was secreted by the microneme and its secretion was regulated by temperature.The anti-rEt AMA2 polyclonal antibodies had no signifi cant effect on sporozoite invasion into host cells.After transient transfection of Et AMA2 into DF-1 cells,the sporozoite invasion rate increased signifi cantly.These results would help further elucidate the function and mechanism of Et AMA2 in Eimeria invasion.
作者
王青杰
朱顺海
赵其平
黄兵
韩红玉
于钰
梁珊珊
余水兰
王海霞
董辉
WANG Qingjie;ZHU Shunhai;ZHAO Qiping;HUANG Bing;HAN Hongyu;YU Yu;LIANG Shanshan;YU Shuilan;WANG Haixia;DONG Hui(Key Laboratory of Animal Parasitology of Ministry of Agriculture and Rural Affairs,Shanghai Veterinary Research Institute,CAAS,Shanghai 200241,China;College of Life and Environment Sciences,Shanghai Normal University,Shanghai 200234,China)
出处
《中国动物传染病学报》
CAS
北大核心
2022年第5期8-18,共11页
Chinese Journal of Animal Infectious Diseases
基金
国家自然科学基金(31672551)
国家寄生虫种质资源共享服务平台(平台-TDRC-22)。
关键词
球虫
柔嫩艾美耳球虫
顶膜抗原2
微线体
入侵
Coccidia
Eimeria tenella
apical membrane antigen 2
microneme
invasion
