嵌合型口蹄疫病毒样颗粒组装研究

Study on the Assembly of Chimeric Virus-like Particles of Foot and Mouth Disease Virus

[目的]为研究和制备多联或多价口蹄疫病毒样颗粒疫苗奠定基础。[方法]将FLAG序列通过融合PCR技术插入口蹄疫结构蛋白VP1GH-loop可变区,并通过大肠杆菌原核表达技术表达口蹄疫病毒衣壳蛋白VP0、VP3和嵌合型VP1,在体外组装出嵌合FLAG外源多肽的口蹄疫病毒样颗粒。[结果]大肠杆菌表达的口蹄疫衣壳蛋白主要以可溶性存在,在缓冲体系中融合标签被切除衣壳蛋白VP0、VP3和FLAGVP1组装成病毒样颗粒,其大小约25 nm左右。FLAG序列成功插入GH-loop可变区第150-151氨基酸位点之间,外源多肽的插入没有影响衣壳蛋白VP1的空间结构。[结论]口蹄疫loop可变区引入外源抗原是可行的,但外源多肽的大小及理化性质会影响病毒样颗粒的组装效率。

[Objective] The research aimed to lay the foundation for studying and preparing multiple or polyvalent virus-like particle vaccine of foot and mouth disease virus.[Method] FLAG sequence was inserted into variable region of structural protein VP1 GH-loop of foot and mouth disease virus.Capsid proteins VP0,VP3 and chimeric VP1 of foot and mouth disease virus were expressed in Escherichia coli prokaryotic expression system.And the virus-like particles of foot and mouth disease virus with FLAG exogenous polypeptide was assembled in vitro.[Result] The capsid proteins of foot and mouth disease virus expressed by E.coli were mainly dissoluble.The fusion protein was cut in the buffer system,and capsid proteins VP0,VP3 and FLAG VP1 were assembled into virus-like particle （the size of about 25 nm）.FLAG sequence was successfully inserted into 150-151 amino acid sites of variable region of GH-loop.The insertion of exogenous polypeptide didn＇t influence the spatial structure of capsid protein VP1.[Conclusion] It was feasible to insert exogenous antigen into variable region of GH-loop,but the size and physical and chemical characteristics of exogenous polypeptide would influence the assembly efficiency of virus-like particle.