Abstract
Human Noroviruses (HuNoVs) are the main cause of non-bacterial gastroenteritis. Contaminated produce is a main vehicle for dissemination of HuNoVs. In this study, we used an ice nucleation protein mediated surface display system to present the protruding domain of GII.4 HuNoV capsid protein on bacterial surface and used it as a new strategy to explore interaction between HuNoV protein and receptor candidates from romaine lettuce. The surface-displayed HuNoV proteins were confirmed on the surface of the transformed bacteria by an immunofluorescence assay. The distribution patterns of the surface-displayed HuNoV proteins in romaine lettuce were identified through a confocal immunofluorescence assay. The surface-displayed HuNoV proteins could be found in the stomata, and the surfaces of vein and leaf of romaine lettuce. The surface-displayed HuNoV proteins could be captured by an ELISA assay utilizing extract from leaf (LE) or vein (VE). The binding of the surface-displayed HuNoV proteins to LE or VE could be competitively blocked by histo-blood group antigens from human saliva. In addition, the binding of the surface-displayed HuNoV proteins to LE or VE could also be attenuated by heat denaturation of lettuce proteins, and abolished by oxidation of lettuce carbohydrates. The results indicated that histo-blood group antigen-like molecules in LE or VE were involved in the binding of the surface-displayed HuNoV proteins to romaine lettuce. All data demonstrated that the surface-displayed HuNoV proteins could be utilized in a new and simple system for investigation of the interaction between the HuNoVs and their candidate ligands.
Highlights
Norovirus (NoV) is a single-stranded RNA virus of the family Caliciviridae
SD-GII.4P was Presented at the Surface of the Transformed E. coli BL21
No FITC fluorescence signals were observed in P (GII.4) BL21 (Figures 2C,D), ice nucleation protein (INP) BL21 samples (Figures 2E,F) and E. coli BL21
Summary
Norovirus (NoV) is a single-stranded RNA virus of the family Caliciviridae. Based on the sequence of the major capsid protein (VP1), noroviruses have been categorized into seven genogroups (GI though GVII) (Vinjé, 2015). The GI, GII, and GIV genogroups are capable of infecting humans, and comprise the human noroviruses (HuNoVs). GII strains are more common, and are the main cause of human non-bacterial gastroenteritis worldwide (Hoa Tran et al, 2013). Infection by HuNoVs could be associated with the consumption of fresh produce, such as romaine lettuce (Ethelberg et al, 2010), onion (Dicaprio et al, 2015), strawberries (Made et al, 2013), and raspberries (Sarvikivi et al, 2012). Harvest and post-harvest cross-contamination of produce could result from human contact, including handling, chopping/slicing and mixing (Grove et al, 2015). Contaminated produce becomes one of the important vehicles for the transmitting of HuNoVs
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