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.
2.Drug screening model of treating pigmentation disorders in tyrp1a transgenic zebrafish
Li LIU ; Siran PEI ; Huali WU ; Qingshun ZHAO ; Jixian PENG ; Jing SHANG
Journal of China Pharmaceutical University 2016;47(6):740-743
To effectively screen treating pigmentation disorders drug, a new transgenic zebrafish drug screening model was constructed. Green fluorescent protein(GFP)were drived by tyrosinase-related protein 1a (tyrp1a)promoter specific expression in melanocyte. Effect of N-Phenylthiourea(PTU)and alpha-melanaocyte stimulating hormone(α-MSH)on the GFP expression and melanogenic of tyrp1a: eGFP zebrafish were photographed under the steromicroscope. Results showed that PTU could significantly inhibit the melanogenic and expression of GFP of zebrafish. As compared with control group, α-MSH treatment resulted in marked stimulation of body pigmentation and expression of GFP. In conclusion, a new transgenic zebrafish drug screening model was successfully established, which can be used for treating pigmentation disorders.