中国动物传染病学报2018，Vol.26Issue(2)：40-48,9.
柔嫩艾美耳球虫伴侣蛋白TCP-1亚基α的特性和功能初步分析
MOLECULAR CHARACTERIZATION AND FUNCTIONAL ANALYSIS OF TCP-1 SUBUNIT α OF EIMERIA TENELLA
摘要
Abstract
Eimeria tenella is an important intracellular parasite that invades cecal epithelial cells of chickens,causing serious coccidiosis. At present, prevention and treatment mainly rely on anti-coccidiosis drugs. As a result, the widespread use of drug causes severe resistance to E.tenella.In our previous study,we analysized differentially expressed genes between two drug-resistant strains(diclazuril-resistant and maduramicin-resistant strains) and drug-sensitive strain of E.tenella through RNA sequencing.TCP-1 subunit α of E. tenella,one of the differentially expressed genes,was up-regulated in diclazuril-resistant and maduramicin-resistant strains.In this study, E.tenella TCP-1 subunit α(Et TCP-1α)gene was cloned in PCR and ligated into the prokaryotic expression vector pET-28a to construct the recombinant plasmid pET-Et TCP-1α.The recombination protein r Et TCP-1α was expressed after induction with IPTG and then used to immunize New Zealand rabbits and BALB/C mouse to prepare polyclonal antibodies. Quantitative real time PCR and Western blot were applied to analyze the mRNA transcription and translation levels of TCP-1 subunit α in resistant and sensitive strains and test the expression levels of TCP-1 subunit α in four developmental stages of E.tenella.The results revealed that Et TCP-1α was expressed at significantly higher levels in the two drug-resistant strains than drug-sensitive strains and the highest expression occurred in the second generation merozoite stage.The protein Et TCP-1α was mainly located in the surfaces of sporozoites and the second generation merozoites as shown in IFA.After the sporozoites invaded DF-1 cells,Et TCP-1α was uniformly distributed in the immature and mature schizonts and its expression increased significantly.Therefore,it was presumed that Et TCP-1α played a certain role during the growth in the host cells and development of drug resistance of E.tenella.关键词
柔嫩艾美耳球虫/TCP-1亚基α/耐药性Key words
Eimeria tenella/TCP-1 subunit α/resistance分类
农业科技引用本文复制引用
陈婷,黄兵,赵其平,董辉,朱顺海,赵宗平,吕凌,严茗,韩红玉..柔嫩艾美耳球虫伴侣蛋白TCP-1亚基α的特性和功能初步分析[J].中国动物传染病学报,2018,26(2):40-48,9.基金项目
中央级公益性科研院所基本科研业务费专项(2016JB10) （2016JB10）
国家寄生虫种质资源共享服务平台(平台-TDRC-22) （平台-TDRC-22）
上海市闵行区人才发展专项基金 （）
