宿主GAPDH对柔嫩艾美耳球虫子孢子入侵细胞的影响

doi:10.11843/j.issn.0366-6964.2019.10.015

Abstract

Abstract: At present, the mechanism of how the host cell responds to Eimeria tenella infection is highly limited. In our previous study, isobaric tags for relative and absolute quantitation were used to study the proteomics changes in host cells after infection with E. tenella sporozoites. In this study, one of the up-regulated proteins, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), was selected to study its effect on the invasion of Eimeria tenella sporozoites. Gallus gallus GAPDH gene was cloned, and the prokaryotic recombinant expression plasmid pGEX-4T-1-GAPDH was constructed. The recombinant GAPDH protein (rGAPDH) was expressed and identified by Western blot. Expression of GAPDH in DF-1 cells during E. tenella sporozoites infection was detected by q-PCR, Western blot and immunohistochemistry. The effect of GAPDH polyclonal antibody on sporozoites invasion of cells was detected by antibody inhibition assay. Results were as follows:A 1 123-bp GAPDH was cloned and the rGAPDH was recognized as a~61.7 kDa protein by Western blot. The expression of GAPDH in E. tenella sporozoites-infected DF-1 cells were up-regulated by q-PCR and immunohistochemistry detection. The antibody inhibition assay showed that antibodies against GAPDH at 50, 100, 200, 300 and 400 μg·mL^-1 could significantly inhibit sporozoite invasion compared with the same does of normal rabbit IgG. These results show that E. tenella sporozoites infection could induce the up-regulation of host GAPDH, and host GAPDH plays an active role in Eimeria invasion.

CLC Number:

S852.723

WANG Lu, ZHU Shunhai, ZHAO Qiping, HUANG Bing, HAN Hongyu, LIU Guiling, LI Zhihang, ZHAO Huanzhi, DONG Hui. Effects of Host Glyceraldehyde-3-phosphate Dehydrogenase on Eimeria tenella Sporozoites Invasion of Cells[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2019, 50(10): 2088-2096.

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Effects of Host Glyceraldehyde-3-phosphate Dehydrogenase on Eimeria tenella Sporozoites Invasion of Cells

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