柔嫩艾美耳球虫延伸因子1α特性与功能初步分析

doi:10.11843/j.issn.0366-6964.2020.12.020

Abstract

Abstract: Coccidiosis infected by Eimeria spp. has a major economic impact on the poultry industry worldwide, resulting in high mortality, low growth, low productivity, and high control costs. Drug resistance of Eimeria spp. is a serious problem with the widespread use of anticoccidial drugs. The focus of coccidiosis prevention and control tends to research on subunit vaccines that are convenient, safe, and easier to produce than live vaccines. Therefore, new methods are urgently needed to effectively control coccidiosis, including the search for specific molecular targets. In this study, cDNA of E. tenella sporulated oocysts (SO) as a template, we amplified the ORF sequences of the elongation factor 1α of Eimeria tenella (EtEF1α). Bioinformatics analyses shows that the gene encodes 450 amino acids with a predicted molecular mass of 49.1 kDa. The isoelectric points were 4.7. Sequence analysis found that the proteins encoded by EtEF1α have no signal peptides and transmembrane domains, but contain N-myristoylation sites. The transcription and translation levels were analyzed using real-time quantitative PCR and Western blot. The results showed that the mRNA transcription levels of EtEF1α were higher in the sporozoite (Spz) and merozoite (Mrz) than unsporulated oocysts (UO) and SO, and the translation levels were high in UO and Mrz. Indirect immunofluorescence localization showed EtEF1α was mainly located at anterior of Spz and the whole cytoplasm of Mrz. As the parasites developed in the cell, the fluorescence intensity gradually brightens. The secretion assay indicated that EtEF1α was a secretion protein, but not secreted from micronemes. Invasion inhibition assay showed that rabbit anti-rEtEF1α polyclonal antibodies can inhibit sporozoite invasion of DF-1 cells. These results indicated that EtEF1α may play a role in the growth and development of the parasite in the host cell and participate in the process of sporozoite invasion into the host cell.

Key words: Eimeria tenella, elongation factor 1α, characteristic analyse, invasion

CLC Number:

S852.723

LIANG Shanshan, ZHU Shunhai, ZHAO Qiping, HUANG Bing, DONG Hui, YU Yu, WANG Qingjie, YU Shuilan, WANG Haixia, HAN Hongyu. Preliminary Study on the Function of Elongation Factor 1α of Eimeria tenella[J]. Acta Veterinaria et Zootechnica Sinica, 2020, 51(12): 3111-3121.

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Preliminary Study on the Function of Elongation Factor 1α of Eimeria tenella

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