Expression of the major capsid protein VP6 of group C rotavirus and synthesis of chimeric single-shelled particles by using recombinant baculoviruses.
VP6 of group C (Cowden strain) rotavirus was expressed in the baculovirus system. The recombinant protein, expressed to a high level in insect cells, was purified by ion-exchange chromatography. The purified protein was proven to be trimeric. The effect of pH on the trimer's stability was investigated. Coexpression of VP6 from group A (bovine strain RF) and VP6 from group C in the baculovirus system did not result in the formation of chimeric trimers. Coexpression of VP2 from group A rotavirus (bovine strain RF) and VP6 from group C in the baculovirus system led to the formation of chimeric, empty, single-shelled particles. These results demonstrate conservation in the domains necessary for binding to VP2 in different serogroups of VP6. The locations of the domains involved in trimerization and in the interaction with VP2 are discussed.
Tosser
G
G
Unité de Virologie et d'Immunologie Moléculaires, C.R.J., Institut National de la Recherche Agronomique, Jouy-en-Josas, France.
Labbé
M
M
Brémont
M
M
Cohen
J
J
D000818:Animals; D000956:Antigens, Viral; D016367:Baculoviridae / Q000235:genetics*; D001483:Base Sequence; D015153:Blotting, Western; D002213:Capsid / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D036022:Capsid Proteins; D002478:Cells, Cultured; D002678:Chimera; D002852:Chromatography, Ion Exchange; D004247:DNA; D004591:Electrophoresis, Polyacrylamide Gel; D005814:Genes, Viral; D006863:Hydrogen-Ion Concentration; D007313:Insecta; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D012401:Rotavirus / Q000378:metabolism*; D014771:Virion / Q000235:genetics* / Q000378:metabolism
D000956:Antigens, Viral; D036022:Capsid Proteins; D011994:Recombinant Proteins; C061332:VP6 protein, Rotavirus; D004247:DNA