title: Budding of peste des petits ruminants virus-like particles from insect cell membrane based on intracellular co-expression of peste des petits ruminants virus M, H and N proteins by recombinant baculoviruses.
abstract: Peste des petits ruminants virus (PPRV), an etiological agent of peste des petits ruminants (PPR), is classified into the genus Morbillivirus in the family Paramyxovirida. In this study, two full-length open reading frames (ORF) corresponding to the PPRV matrix (M) and haemagglutinin (H) genes underwent a codon-optimization based on insect cells, respectively. Two codon-optimized ORFs along with one native nucleocapsid (N) ORF were used to construct recombinant baculoviruses co-expressing the PPRV M, H and N proteins in insect cells. Analysis of Western blot, immunofluorescence, confocal microscopy and flow cytometry demonstrated co-expression of the three proteins but at different levels in insect cells, and PPR virus-like particles (VLPs) budded further from cell membrane based on self-assembly of the three proteins by viewing of ultrathin section with a transmission electron microscope (TEM). Subsequently, a small number of VLPs were purified by sucrose density gradient centrifugation for TEM viewing. The PPR VLPs, either purified by sucrose density gradient centrifugation or budding from insect cell membrane on ultrathin section, morphologically resembled authentic PPRVs but were smaller in diameter by the TEM examination.
authors.affiliation: National Research Center for Exotic Animal Diseases, China Animal Health and Epidemiology Center, No. 369 Nanjing Road, Qingdao 266032, Shandong, China.
authors.forename: Fuxiao
authors.initials: F
authors.lastname: Liu
authors.affiliation: National Research Center for Exotic Animal Diseases, China Animal Health and Epidemiology Center, No. 369 Nanjing Road, Qingdao 266032, Shandong, China.
authors.forename: Xiaodong
authors.initials: X
authors.lastname: Wu
authors.affiliation: National Research Center for Exotic Animal Diseases, China Animal Health and Epidemiology Center, No. 369 Nanjing Road, Qingdao 266032, Shandong, China.
authors.forename: Yonggang
authors.initials: Y
authors.lastname: Zhao
authors.affiliation: National Research Center for Exotic Animal Diseases, China Animal Health and Epidemiology Center, No. 369 Nanjing Road, Qingdao 266032, Shandong, China.
authors.forename: Lin
authors.initials: L
authors.lastname: Li
authors.affiliation: National Research Center for Exotic Animal Diseases, China Animal Health and Epidemiology Center, No. 369 Nanjing Road, Qingdao 266032, Shandong, China. Electronic address: wangzhiliang@cahec.cn.
authors.forename: Zhiliang
authors.initials: Z
authors.lastname: Wang
mesh_terms: D000818:Animals; D016367:Baculoviridae / Q000235:genetics*; D002462:Cell Membrane / Q000648:ultrastructure / Q000821:virology*; D002499:Centrifugation, Density Gradient; D005822:Genetic Vectors; D006389:Hemagglutinins, Viral / Q000235:genetics / Q000378:metabolism*; D046529:Microscopy, Electron, Transmission; D009698:Nucleoproteins / Q000235:genetics / Q000378:metabolism*; D018107:Peste-des-petits-ruminants virus / Q000235:genetics* / Q000502:physiology; D061987:Sf9 Cells; D018411:Spodoptera; D014763:Viral Matrix Proteins / Q000235:genetics / Q000378:metabolism*; D022701:Virosomes / Q000378:metabolism / Q000648:ultrastructure; D019065:Virus Assembly; D057074:Virus Release
chemical_list: D006389:Hemagglutinins, Viral; D009698:Nucleoproteins; D014763:Viral Matrix Proteins; D022701:Virosomes
keywords: Baculovirus; Budding; Co-expression; Insect cell; Peste des petits ruminants virus; Virus-like particle