摘要
目的分析我国GⅡ.4型诺如病毒(norovirus,NoV)GZ19株的进化特征,并明确其结合组织血型抗原(histoblood group antigens,HBGAs)受体的能力和方式。方法根据GZ19株中的ORF2区序列,构建进化树,并分析其在HBGAs结合位点(HBGA binding sites,HBSs)和关键阻断表位的氨基酸序列。采用原核表达系统表达P颗粒并进行纯化,获得的蛋白经SDS-PAGE和间接ELISA法鉴定后,采用唾液结合试验和寡糖结合试验分析P颗粒的糖结合特征。结果GZ19株属于GⅡ.4 Sydney[P31]谱系,其受体结合位点和阻断表位的氨基酸序列相对保守,与近5年其他GⅡ.4Sydney[P31]毒株具有较高的同源性,而与GⅡ.4 Sydney 2012原型株和GⅡ.4 Sydney[P16]株的差异较大。P颗粒仅与A、B、O、AB分泌型唾液和H-di寡糖结合。结论GZ19株代表目前GⅡ.4 Sydney[P31]NoV的进化方向,P颗粒的成功表达及其与HBGAs受体的结合特征分析,为研究我国GⅡ.4 NoVs的流行进化规律及疫苗开发奠定了基础。
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.
作者
王金冬
马亚林
段招军
WANG Jindong;MA Yalin;DUAN Zhaojun(Department of Pathogen Biology,School of Basic Medicine,Weifang Medical University,Weifang 261053,Shandong Province,China;不详)
出处
《中国生物制品学杂志》
CAS
CSCD
北大核心
2023年第5期531-536,544,共7页
Chinese Journal of Biologicals
基金
国家科技重大专项(2018ZX10305409-004-002)。
