title: Generation of Sendai virus nucleocapsid-like particles in yeast.
abstract: The gene encoding Sendai virus nucleocapsid protein was cloned into the yeast Saccharomyces cerevisiae expression vector pFGG3 under control of GAL7 promoter. The high level of recombinant Sendai virus nucleocapsid protein expression (12-14 mg/l of yeast culture) was obtained. The evaluation of recombinant proteins expression in yeast by Western blot analysis revealed specific reactivity with immune sera. Electron microscopy demonstrated the assembly of typical herring-bone structures of purified recombinant nucleocapsid protein. These structures contained host RNA, which was resistant to an RNase treatment. The nucleocapsid protein revealed stability in yeast and can be easily purified by cesium chloride gradient ultracentrifugation. The development of a simple, efficient and cost-effective system for generation of Sendai virus nucleocapsid protein might help to upgrade reagents for virus serology, and facilitate investigation of virus replication and RNA encapsidation mechanisms.
authors.affiliation: Institute of Biotechnology, V.Graiciuno 8, Vilnius LT-2028, Lithuania. jmind@ibt.lt
authors.forename: Mindaugas
authors.initials: M
authors.lastname: Juozapaitis
authors.forename: Rimantas
authors.initials: R
authors.lastname: Slibinskas
authors.forename: Juozas
authors.initials: J
authors.lastname: Staniulis
authors.forename: Takemasa
authors.initials: T
authors.lastname: Sakaguchi
authors.forename: Kestutis
authors.initials: K
authors.lastname: Sasnauskas
mesh_terms: D003001:Cloning, Molecular; D005814:Genes, Viral; D046911:Macromolecular Substances / Q000378:metabolism; D008854:Microscopy, Electron; D019590:Nucleocapsid Proteins / Q000096:biosynthesis* / Q000235:genetics; D012331:RNA, Fungal / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism*; D029082:Sendai virus / Q000235:genetics*; D019065:Virus Assembly
chemical_list: D046911:Macromolecular Substances; D019590:Nucleocapsid Proteins; D012331:RNA, Fungal; D011994:Recombinant Proteins