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@#Objective To analyze the evolutionary characteristics of GZ19 strain of G Ⅱ.4 norovirus(NoV) in China,and clarify its ability and mode of binding to receptors of histo-blood group antigens(HBGAs).Methods According to the sequence of ORF2 region in GZ19 strain,the evolutionary tree was constructed and the amino acid sequences at HBGA binding sites(HBSs) and key blocking epitopes were analyzed.P particles were expressed by prokaryotic expression system and purified.The obtained protein was identified by SDS-PAGE and indirect ELISA,and analyzed for the receptor binding characteristics of P particles by saliva binding and oligosaccharide binding assays.Results The GZ19 strain belonged to G Ⅱ.4Sydney [P31] lineage,of which the amino acid sequences of receptor binding sites and blocking epitopes were relatively conservative.It showed high homology with other G Ⅱ.4 Sydney [P31] strains in recent five years,while significant difference from G Ⅱ.4 Sydney 2012 original strain and G Ⅱ.4 Sydney [P16] strains.P particles only combined with A,B,O,AB secretory saliva and H-di oligosaccharide.Conclusion GZ19 strain represented the current evolutionary direction of G Ⅱ.4Sydney [P31] NoV.The successful expression of P particles and analysis of the binding characteristics with HBGA receptors laid a foundation of the research of epidemic evolution dynamics and vaccine development of G Ⅱ.4 NoVs in China.
RESUMO
Objective@#The gene engineering technique was used to express the P[6] genotype rotavirus (rotaviruses, RVs) GST-VP8*-Z84 protein from pigs, and the binding characteristics of the protein to oligosaccharide and salivary receptor were studied.@*Methods@#The GST-VP8*-Z84 protein was purified by GST Escherichia coli expression system and affinity chromatography using porcine P[6]. Enzyme-linked immunosorbent assay (ELISA) saliva binding test and oligosaccharide binding test were used to analyze the binding characteristics of the genotype to receptors.@*Results@#Porcine P[6] GST-VP8*-Z84 protein bound well to mucin core 2.@*Conclusions@#The potential receptor of P[6] RV may be the core of mucin, which may provide the experimental basis and theoretical basis for the mechanism of rotavirus and receptor interaction and the development of RV vaccine and highly effective therapeutic drugs.
RESUMO
Objective@#During the 2016 winter season, GII.2 norovirus(NoV) suddenly emerged in China. To elucidate its mechanism of epidemic, this study focused on characteristics of binding between the P protein of capsid and histo-blood group antigens (HBGAs).@*Methods@#The research object was GⅡ.2 ZTX strain which had an outbreaks by the end of 2016 in Beijing. Recombinant prokaryotic expression plasmid was constructed, and the expression of virus P protein was determined and purified. The P protein characteristics of binding to HBGAs was studied through saliva and oligosaccharide binding experiments.@*Results@#Soluble P protein was successfully obtained, and combined with type A, B, AB saliva.@*Conclusions@#The result illuminate the combination with new outbreaks of NoV and salivary types, which provided a basis for its pathogenic mechanism and prevention and control measures.