Expression of GⅡ-4 strain NoV virus-like particles and analysis of its binding activity with HBGAs receptor

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Expression of GⅡ-4 strain NoV virus-like particles and analysis of its binding activity with HBGAs receptor

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- Expression of GⅡ-4 strain NoV virus-like particles and analysis of its binding activity with HBGAs receptor
- Journal of Chongqing Medical University Vol. 36, Issue 3, Pages: 315-318(2011)
- 作者机构：
  
  1. 南方医科大学中医药学院中医基础理论教研室
  2. 南方医科大学公共卫生与热带医学学院流行病学系
- 作者简介：
- 基金信息：
- DOI：10.13406/j.cnki.cyxb.2011.03.006
  CLC： R392
- Published：2011，
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[1]张绪富,戴迎春,宋灿磊,周迎春.诺如病毒病毒样颗粒的表达及其与HBGAs受体的结合研究[J].重庆医科大学学报,2011,36(03):315-318.

ZHANG Xu-fu, et al. Expression of GⅡ-4 strain NoV virus-like particles and analysis of its binding activity with HBGAs receptor. [J]. Journal of Chongqing Medical University 36(3):315-318(2011)
[1]张绪富,戴迎春,宋灿磊,周迎春.诺如病毒病毒样颗粒的表达及其与HBGAs受体的结合研究[J].重庆医科大学学报,2011,36(03):315-318. DOI： 10.13406/j.cnki.cyxb.2011.03.006.

ZHANG Xu-fu, et al. Expression of GⅡ-4 strain NoV virus-like particles and analysis of its binding activity with HBGAs receptor. [J]. Journal of Chongqing Medical University 36(3):315-318(2011) DOI： 10.13406/j.cnki.cyxb.2011.03.006.

摘要

目的:以杆状病毒为载体

在昆虫细胞sf9中表达GⅡ-4型野生株诺如病毒（Noroviruses

NoV）GZ121的衣壳蛋白ORF2

并验证其与组织血型抗原（Histoblood group antigen

HBGAs）受体的结合活性。方法:双酶切质粒PMD18-T-GZ121 ORF2酶切

连接入线性化载体PFastBacTM1

获得重组转座载体pFastBac-GZ121 ORF2

利用Bac-to-Bac杆状病毒表达系统获得重组杆状病毒

将其转染昆虫sf9细胞进行蛋白表达

用蔗糖超离方法纯化以及免疫印迹、透射电镜等方法鉴定表达产物

用EIA法检测NoV-VLP与HBGAs的结合特性。结果:成功表达了分子量约为58 kD的NoV衣壳蛋白

大小约30 nm与预期相符

NoV-VLP与A、B、O型均能结合

与非分泌型唾液不结合

结合方式与rVA387相一致。结论:我国优势流行株GⅡ4型NoV病毒样颗粒（Virus-like particles

VLP）的表达成功及NoV-VLP和HBGAs受体结合模型的建立

为我国NoV疫苗的开发和防治药物的研究奠定了基础。

Abstract

Objective: To express the capsids of the predominant strain NoV(GⅡ-4) of our country and explore its binding activity and pattern model with HBGAs receptor.Methods: GZ121 NoV ORF2 gene was constructed into baculovirus expression system.After the transfection to sf9 cells

the recombinant proteins were purified by sucrose-ultra centrifugation

and then were identified by SDS-PAGE

Western blot and TEM.And the binding patterns of NoV to HBGAs antigens were identified by EIA.Results: VLPs expression was confirmed by using SDS-PAGE

Western blot and could bind to all secretors(OD>2.5)

but not to all nonsecretors(OD≈0.3).Conclusion: The GⅡ-4 NoV VLP was successfully expressed and HBGAs receptor binding assays were established.These data will help us to explore the host adaptation character of NoV and apply the assays to screen drugs for antivirals against Noroviruses.The results obtained in the present work will also lay a foundation for the development of the immunological diagnostic reagents as well as vaccines for Norovirus.

关键词

诺如病毒杆状病毒表达系统病毒样颗粒HBGAs受体

Keywords

norovirusbaculovirus expression vector systemvirus-like particlesHBGAs receptor

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