title: [Preparation of human parvovirus B19 virus-like particles].
abstract: The baculovirus expression system was employed to prepare the virus-like particles (VLPs) of human parvovirus B19. The synthesized VP2 gene of B19 was inserted into the multi-cloning site (MCS) of pFastBac1 vector; the resulting plasmid was transferred to the Escherichia coli DH10Bac competent cells, which contain a baculovirus shuttle vector (Bacmid), to generate Bacmid-VP2 by site-specific transposition. Recombinant baculovirus carrying VP2 gene (rBac-VP2) was then rescued from Bacmid-VP2-transfected Sf9 cells. Indirect immunofluorescence and Western blotting were used to identify the VP2 protein in rBac-VP2-infected Sf9 cells, and the VLPs were observed under transmission electron microscope after being enriched by ultracentrifugation. The B19 VLPs were successfully produced in insect cells with baculovirus expression system, which will facilitate the development of diagnostic reagents to detect the antibody against B19 virus in human serum.
authors.affiliation: Department of Microbiology, School of Medicine, Shandong University, Jinan 250012, China.
authors.forename: Xiaohui
authors.initials: X
authors.lastname: Zou
authors.forename: Liuxin
authors.initials: L
authors.lastname: Dong
authors.forename: Jingdong
authors.initials: J
authors.lastname: Song
authors.forename: Jianguo
authors.initials: J
authors.lastname: Qu
authors.forename: Xiuping
authors.initials: X
authors.lastname: Yu
authors.forename: Zhuozhuang
authors.initials: Z
authors.lastname: Lu
authors.forename: Tao
authors.initials: T
authors.lastname: Hong
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000097:blood; D016367:Baculoviridae / Q000235:genetics / Q000378:metabolism*; D036022:Capsid Proteins / Q000096:biosynthesis* / Q000235:genetics; D002460:Cell Line; D003001:Cloning, Molecular; D005822:Genetic Vectors / Q000235:genetics; D016732:Parvovirus B19, Human / Q000235:genetics* / Q000276:immunology; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification; D014771:Virion / Q000235:genetics / Q000378:metabolism*
chemical_list: D000914:Antibodies, Viral; D036022:Capsid Proteins; D011994:Recombinant Proteins; C095548:capsid protein VP2, parvovirus B19