Generation of Tioman virus nucleocapsid-like particles in yeast Saccharomyces cerevisiae

• Published in: Virus research Vol. 145; no. 1; pp. 92 - 96
• Main Authors: Petraityte, Rasa, Tamosiunas, Paulius L., Juozapaitis, Mindaugas, Zvirbliene, Aurelija, Sasnauskas, Kestutis, Shiell, Brian, Russell, Gail, Bingham, John, Michalski, Wojtek P.
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01-10-2009
• Subjects: Amino Acid Sequence; Animals; Antibodies, Viral - immunology; Antigens, Viral - immunology; Chiroptera; Expression; Immunohistochemistry; Mice; Microscopy, Electron, Transmission; Molecular Sequence Data; Nipah virus; Nucleocapsid; Nucleocapsid Proteins - biosynthesis; Nucleocapsid Proteins - genetics; Nucleocapsid Proteins - immunology; Nucleocapsid Proteins - ultrastructure; Nucleocapsid-like particles; Paramyxoviridae; Paramyxoviridae Infections - immunology; Paramyxoviridae Infections - virology; Paramyxovirus; Pneumovirinae - genetics; Pneumovirinae - immunology; Pneumovirinae - isolation & purification; Pteropus; Rabbits; Recombinant Proteins - biosynthesis; Recombinant Proteins - genetics; Recombinant Proteins - immunology; Recombinant Proteins - ultrastructure; Saccharomyces cerevisiae; Saccharomyces cerevisiae - metabolism; Swine; Tioman virus; Tioman virus; Immunohistochemistry; Nucleocapsid-like particles; Expression; Nucleocapsid
• ISSN: 0168-1702; 1872-7492
• DOI: 10.1016/j.virusres.2009.06.013

Tioman virus (TioV) was isolated from a number of pooled urine samples of Tioman Island flying foxes (
Pteropus hypomelanus) during the search for the reservoir host of Nipah virus. Studies have established TioV as a new virus in the family
Paramyxoviridae. This novel paramyxovirus is antigenically related to Menangle virus that was isolated in Australia in 1997 during disease outbreak in pigs. TioV causes mild disease in pigs and has a predilection for lymphoid tissues. Recent serosurvey showed that 1.8% of Tioman Islanders had neutralizing antibodies against TioV, indicating probable past infection. For the development of convenient serological tests for this virus, recombinant TioV nucleocapsid (N) protein was expressed in the yeast
Saccharomyces cerevisiae. High yields of recombinant TioV N protein were obtained. Electron microscopy demonstrated that purified recombinant N protein self-assembled into nucleocapsid-like particles which were identical in density and morphology to authentic nucleocapsids from paramyxovirus-infected cells. Different size nucleocapsid-like particles were stable and readily purified by CsCl gradient ultracentrifugation. Polyclonal sera raised in rabbits after immunization with recombinant TioV N protein reacted reliably with TioV infected tissues in immunohistochemistry tests. It confirmed that the antigenic properties of yeast derived TioV N protein are identical to authentic viral protein.