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An Effective Platform for Exploring Rotavirus Receptors by Bacterial Surface Display System

Virologica Sinica 2020年第1期35卷 103-109页

作者： Danlei Liu Haoran Geng Zilei Zhang Yifan Xing Danlu Yang Zhicheng Liu Dapeng Wang Department of Food Science and Technology School of Agriculture and BiologyShanghai Jiao Tong UniversityShanghai 200240China State Key Laboratory of Applied Microbiology Southern China Guangdong Provincial Key Laboratory of Microbial Culture Collection and ApplicationGuangdong Open Laboratory of Applied MicrobiologyGuangdong Institute of MicrobiologyGuangzhou 510070China Shanghai Food Safety and Engineering Technology Research Center School of Agriculture and BiologyShanghai Jiao Tong UniversityShanghai 200240China

Rotavirus(RV) is a major foodborne pathogen. For RV prevention and control, it is a key to uncover the interaction mechanism between virus and its receptors. However, it is hard to specially purify the viral receptors, including histo-blood group antigens(HBGAs). Previously, the protruding domain protein(P protein) of human norovirus(genotype Ⅱ.4) was displayed on the surface of Escherichia coli, and it specifically recognized and captured the viral ligands. In order to further verify the feasibility of the system, P protein was replaced by VP8* of RV(G9 P[8]) in this study. In the system, VP8*could be correctly released by thrombin treatment with antigenicity retaining, which was confirmed by Western blot and Enzyme-Linked Immunosorbent Assays. Type A HBGAs from porcine gastric mucin(PGM) were recognized and captured by this system. From saliva mixture, the captured viral receptor bound with displayed VP8* was confirmed positive with monoclonal antibody against type A HBGAs. It indicated that the target ligands could be easily separated from the complex matrix. These results demonstrate that the bacterial surface display system will be an effective platform to explore viral receptors/ligands from cell lines or food matrix.

关键词： Rotavirus Bacterial surface display system Receptors histo-blood group antigens(HBGAs) Saliva

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