1.The research of genotype 4 hepatitis E virus(HEV) capsid recombinant protein and the construction of cellular model for the investigation of viral infection early period
Min ZHAO ; Jingxian LI ; Zizheng ZHENG ; Qingshun GUO ; Hui HUANG ; Wangsheng LAI ; Ji MIAO ; Shengxiang GE ; Jun ZHANG ; Ningshao XIA
Chinese Journal of Microbiology and Immunology 2010;30(8):743-748
Objective To express the recombinant caspid of genotype 4 hepatitis E virus(HEV) ORF2. Methods HEV recombinant capsid protein D66 was expressed in E. coli, using the ORF2 fragment (aa368-606, obtained from swine bile) of genotype 4 HEV. Results The recombinant capsid proteins D66 self-assemble to be particle with a radius of 13 nm through dimeric form in neutral solution. Coated particles reacted well with sera obtained from patients during acute or recovered phase of HEV infection. Immunofluorescence and immnoblot assay suggested that D66 bound and penetrated HepG2 cell lines, and the process of attachment was blocked by sera collected from patients during acute or recovered phase of HEV infection.Conclusion Recombinant D66 particles simulate the structure at the surface of genotype 4 HEV well and specifically adhere and penetrate the host cells, which lays the foundation for the investigation of the molecular mechanism of genotype 4 HEV infection.
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