柔嫩艾美耳球虫含HD结构域蛋白特性和功能初步研究

doi:10.11843/j.issn.0366-6964.2022.08.019

Abstract

Abstract: Coccidiosis is a serious intestinal parasitic disease caused by Eimeria spp., which causes huge economic losses to poultry industry all over the world every year. At present, coccidiosis is mainly controlled by anti-coccidial drugs. Drug resistance of Eimeria to all of used drugs has been developed due to the long-term and unreasonable use of drugs. To study the molecular mechanism of the resistance of Eimeria to drugs, differentially expressed genes between two drug-resistant strains (diclazuril-resistant strains, maduramicin-resistant strains) and drug-sensitive strain of Eimeria tenella were obtained by using RNA sequncing in our lab. We found that E.tenella contained HD domain protein (EtHDCP) up-regulated in two drug-resistant strains. In this paper, EtHDCP gene was successfully cloned from the first cDNA strand of E.tenella drug sensitive strain sporulated oocysts (SO) as template, and the recombinant plasmid pGEX-4T-EtHDCP was constructed, and the recombinant protein rEtHDCP was induced successfully. qRT-PCR and Western blot were used to analyze the transcription and translation levels of EtHDCP in the four different developmental stages of E. tenella drug sensitive strains. At the same time, the translation levels of two drug-resistant strains and drug sensitive of E.tenella were detected. The results showed that the transcription and translation level of EtHDCP in second-generation merozoites stage were the highest, the protein translation level of EtHDCP in drug-resistant strains were significantly higher than those in drug sensitive strains. Indirect immunofluorescence localization showed that the protein was mainly located on the surface of sporozoites and merozoites and in the cytoplasm of merozoites. Invasion inhibition test showed that rabbit anti-rEtHDCP polyclonal antibody could effectively inhibit invasion of sporozoites to host cells. These results showed that the protein may be involved in the growth and development of parasite in host cells, the development of drug resistance and the invasion of sporozoites into host cells.

Key words: Eimeria tenella, HD domain protein, invasion, resistance

CLC Number:

S852.723

XIAO Ke, CHEN Ting, ZHAO Qiping, ZHU Shunhai, DONG Hui, LIU Manyu, YU Yu, HUANG Bing, HAN Hongyu. Preliminary Study on the Characteristics and Functions of HD Dome-containing Proteins in Eimeria tenella[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(8): 2608-2620.

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Preliminary Study on the Characteristics and Functions of HD Dome-containing Proteins in Eimeria tenella

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