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ARS Home » Pacific West Area » Albany, California » Western Regional Research Center » Produce Safety and Microbiology Research » Research » Publications at this Location » Publication #342859
Research Project: Ecology and Detection of Human Pathogens in the Produce Production Continuum

Location: Produce Safety and Microbiology Research

Title: A bacterial surface display system expressing cleavable capsid proteins of human norovirus: a novel system to discover candidate receptors

Author
item XU, QIAN - Shanghai Jiaotong University
item NI, PEIEN - Shanghai Jiaotong University
item LIU, DANLEI - Shanghai Jiaotong University
item YIN, YUJIE - Shanghai Jiaotong University
item LI, QIANQIAN - Shanghai Institute Of Technology
item ZHANG, JYMIE - Guangdong Academy Of Agricultural Sciences
item WU, QIANPING - Guangdong Academy Of Agricultural Sciences
item Tian, Peng
item SHI, XIANMING - Shanghai Jiaotong University
item WANG, DAPENG - Shanghai Jiaotong University

Submitted to: Frontiers in Microbiology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 11/20/2017
Publication Date: 12/6/2017
Citation: Xu, Q., Ni, P., Liu, D., Yin, Y., Li, Q., Zhang, J., Wu, Q., Tian, P., Shi, X., Wang, D. 2017. A bacterial surface display system expressing cleavable capsid proteins of human norovirus: a novel system to discover candidate receptors. Frontiers in Microbiology. 8:2405. https://doi.org/10.3389/fmicb.2017.02405.
DOI: https://doi.org/10.3389/fmicb.2017.02405

Interpretive Summary:

Technical Abstract: Human noroviruses (HuNoVs) are the dominant cause of food-borne outbreaks of gastroenteritis. However, the fundamental of the research on the viruses were restricted by the current immature system to culture HuNoVs and lacking of efficient small animal models. Previously, we demonstrated that the recombinant P domain of HuNoV (GII.4) was successfully anchored on the surface of Escherichia coli BL21 cells after inducing with isopropyl ß-D-1-thiogalactopyranoside (IPTG). The cell surface displayed P proteins could bind to histo-blood group antigens (receptors of HuNoVs). In this study, a linker which contained a thrombin susceptible sequence was added between the bacterial membrane anchor protein and the protruding domain (P protein) of HuNoV (GII.4) capsid proteins. The cleavable bacterial surface displayed system was then used to discover candidate receptors of HuNoV. We demonstrated that the surfaced displayed HuNoV P proteins could be released by thrombin treatment and visualized under electronic microscopy. The bacteria with the surface-displayed HuNoV P proteins were then incubated with pig stomach mucin which contained type O and A HBGAs, followed by a low speed centrifugation to isolate HuNoV P protein-receptor complex. The HuNoV P protein-receptor complex was then released by thrombin treatment. The released viral receptors could be confirmed by using MAbs against correspond HBGAs. The new strategy could be used as a simple approach to characterize unknown receptors for HuNoV in different samples including mammalian cells, oysters, and/or fresh produce.