GI.5和GII.4诺如病毒P蛋白的克隆表达及与长牡蛎类HBGAs的结合特性

Cloning and Expression of Human Norovirus GI.5 and GII.4 P Proteins and Their Binding Characteristics with Histo-Blood Group Antigens-like Substances in Pacific Oysters

为明确人诺如病毒(human norovirus,HuNoV)与长牡蛎类组织血型抗原(histo-blood group antigens,HBGAs)的结合特性,本实验运用大肠杆菌表达系统,克隆表达了基因簇I.5(genogroup I.5,GI.5)和GII.4 HuNoV P蛋白,采用酶联免疫吸附测定研究HuNoV P蛋白与唾液HBGAs和长牡蛎类HBGAs的结合特性。结果表明,GII.4 HuNoV与唾液A型、B型、AB型和O型HBGAs均有较好的结合,而GI.5 HuNoV与B型HBGAs结合较弱,与O型HBGAs具有明显的结合优势。GI.5和GII.4 HuNoV在长牡蛎鳃、消化腺和外套膜中均可富集,其中在消化腺中富集最多,二者主要与类A型和H1型HBGAs结合,GII.4HuNoV与类Lea型、Leb型、Lex型和Ley型HBGAs有不同程度的结合,而GI.5 HuNoV与类Leb型HBGAs仅微弱结合,与类H1型HBGAs具有明显结合优势。综上,不同型别HuNoV与HBGAs的结合特性不尽相同,GII.4HuNoV具有广谱结合特性,GI.5HuNoV具有选择结合特性。

To clarify the binding characteristics of human norovirus(HuNoV)with histo-blood group antigens(HBGAs)-like substances from Pacific oysters,this study used an Escherichia coli expression system to clone and express HuNoV GI.5 and GII.4 P proteins,and analyzed the binding characteristics of HuNoV P proteins with salivary HBGAs and HBGAs-like substances from Pacific oysters using enzyme-linked immunosorbent assay.The results showed that HuNoV GII.4 exhibited good binding characteristics with blood type A,B,AB,and O salivary HBGAs,while GI.5 HuNoV exhibited weak binding characteristics with blood type B salivary HBGAs but had significant advantage in binding with type O salivary HBGAs.HuNoV GI.5 and GII.4 could be bioaccumulated in the gills,digestive gland,and mantle of Pacific oysters,with the highest bioaccumulation in the digestive gland.Both types of HuNoV were mainly bound to type A and H1 HBGAs-like substances;HuNoV GII.4 had different degrees of binding with type Lea,Leb,Lex,and Ley HBGAs-like substances,while HuNoV GI.5 had weak binding with type Leb HBGAs-like substances but significant advantage in binding with type H1 HBGAs-like substances.In summary,different types of HuNoV have different binding characteristics with HBGAs or HBGAs-like substances.Specifically,HuNoV GII.4 shows broad-spectrum binding characteristics whereas HuNoV GI.5 shows selective binding characteristics.