猪丁型冠状病毒S1-CTD相互作用宿主蛋白的筛选和鉴定

doi:10.11843/j.issn.0366-6964.2022.07.022

Abstract

Abstract: The C-terminal domain (CTD) of porcine deltacoronavirus S1 subunit is the main region which induces the neutralizing antibody. S1-CTD was expressed by HEK-293T eukaryotic expression system and purified, and porcine ileal epithelium cells membrane proteins were extracted to investigate porcine host proteins that interact with it. Thirty-two suspected interacting host proteins were obtained by co-inmunprecipitation (Co-IP) and mass spectrometry. Eukaryotic expression plasmid of KIF1 binding protein (KIFBP) was constructed, and the interaction between KIFBP and S1-CTD was identified by Co-IP and laser confocal microscopy. All results proved that KIFBP interacted with S1-CTD and co-located in cytoplasm. Further research indicated that overexpression of KIFBP could effectively reduce the viral mRNA level and the viral titer in which the mRNA level decreased by about 70%, and the viral titer decreased by 10^1.6TCID₅₀. In conclusion, a host protein KIFBP interacting with PDCoV S1-CTD was screened and identified in this study which provides a theoretical basis for understanding the pathogenesis of PDCoV.

Key words: porcine deltacoronavirus, S1-CTD protein, interaction, KIF1 binding protein

CLC Number:

S852.659.6

TANG Rongfeng, FAN Qianjin, GUO Longjun, ZHANG Xin, SHI Da, SHI Hongyan, CHEN Jianfei, FENG Li. Screening and Identification of Host Proteins Interacting with PDCoV S1-CTD[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(7): 2260-2267.

References 18

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Screening and Identification of Host Proteins Interacting with PDCoV S1-CTD

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