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Assembly of double-layered virus-like particles in mammalian cells by coexpression of human rotavirus VP2 and VP6.
- Source :
-
The Journal of general virology [J Gen Virol] 1995 Sep; Vol. 76 ( Pt 9), pp. 2357-60. - Publication Year :
- 1995
-
Abstract
- Development in mammalian cells of a recombinant expression system that mimics the rotavirus capsid assembly process would be advantageous for studying the structural requirements for particle formation. To this end, we investigated the ability of a recombinant vaccinia virus system to produce double-layered virus-like particles. The genes coding for VP2 and VP6 proteins of the human rotavirus strain Wa were cloned and used to generate recombinant vaccinia viruses. Metabolic labelling of CV-1 cells infected with these recombinant viruses followed by immunoprecipitation with a polyclonal antiserum directed to Wa virus showed that VP2 and VP6 were efficiently expressed. The recombinant proteins were similar in size and immunoreactivity to authentic rotavirus proteins. Biochemical and electron microscopy analyses demonstrated that simultaneous expression of VP2 and VP6 in mammalian cells resulted in the formation of intracellular spherical particles resembling double-layered rotavirus particles.
- Subjects :
- Animals
Base Sequence
Capsid genetics
Capsid Proteins
Cell Line
DNA Primers
DNA, Viral
Humans
Mammals
Molecular Sequence Data
Recombinant Fusion Proteins genetics
Rotavirus genetics
Rotavirus ultrastructure
Vaccinia virus genetics
Virion ultrastructure
Virus Replication
Antigens, Viral
Capsid biosynthesis
Rotavirus physiology
Subjects
Details
- Language :
- English
- ISSN :
- 0022-1317
- Volume :
- 76 ( Pt 9)
- Database :
- MEDLINE
- Journal :
- The Journal of general virology
- Publication Type :
- Academic Journal
- Accession number :
- 7561777
- Full Text :
- https://doi.org/10.1099/0022-1317-76-9-2357