Preparation of human parvovirus B19 virus-like particles.
- Author:
Xiaohui ZOU
1
;
Liuxin DONG
;
Jingdong SONG
;
Jianguo QU
;
Xiuping YU
;
Zhuozhuang LU
;
Tao HONG
Author Information
1. Department of Microbiology, School of Medicine, Shandong University, Jinan 250012, China.
- Publication Type:Journal Article
- MeSH:
Animals;
Antibodies, Viral;
blood;
Baculoviridae;
genetics;
metabolism;
Capsid Proteins;
biosynthesis;
genetics;
Cell Line;
Cloning, Molecular;
Genetic Vectors;
genetics;
Parvovirus B19, Human;
genetics;
immunology;
Recombinant Proteins;
biosynthesis;
genetics;
isolation & purification;
Virion;
genetics;
metabolism
- From:
Chinese Journal of Biotechnology
2009;25(4):575-579
- CountryChina
- Language:Chinese
-
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.
