Beet western yellows luteovirus capsid proteins produced by recombinant baculoviruses assemble into virion-like particles in cells and larvae of Bombyx mori.
We subcloned various constructs of the beet western yellows luteovirus (BWYV) 3' proximal genes into Bombyx mori nuclear polyhedrosis virus (BmNPV) transfer vectors and constructed recombinant BmNPVs. The recombinant BmNPVs were used to infect BmN cells and B. mori larvae. Protein expression was analyzed by SDS-PAGE and immunoblot analysis, and the BWYV-encoded capsid protein (CP), P19, readthrough protein (RT), and a modified capsid-readthrough protein (CP-RT*) were detected in BmN cells infected by the respective BmNPVs. However, we detected only the BWYV CP and P19 in BmN cells infected with a recombinant BmNPV containing all three BWYV 3' proximal genes. BmN cells and fat body cells from B. mori larvae infected with recombinant BmNPVs were analyzed by transmission electron microscopy and immunogold labeling. Particles which morphologically and serologically resembled BWYV virions were detected in the nuclei of BmN cells and B. mori larval fat body cells infected with recombinant BmNPVs producing either the BWYV CP alone or BWYV CP plus a BWYV CP-RT*. Similar particles were not detected in cells infected with a control BmNPV or recombinant BmNPV producing only CP-RT*.
D000818:Animals; D016367:Baculoviridae / Q000378:metabolism*; D001483:Base Sequence; D012831:Bombyx / Q000166:cytology / Q000648:ultrastructure / Q000821:virology; D002213:Capsid / Q000096:biosynthesis / Q000502:physiology*; D002460:Cell Line; D017931:DNA Primers / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D015151:Immunoblotting; D007150:Immunohistochemistry; D007814:Larva / Q000648:ultrastructure / Q000821:virology; D017793:Luteovirus / Q000502:physiology* / Q000648:ultrastructure; D016253:Microscopy, Immunoelectron; D008969:Molecular Sequence Data; D010944:Plants / Q000821:virology; D011817:Rabbits; D011994:Recombinant Proteins / Q000096:biosynthesis; D014771:Virion / Q000502:physiology* / Q000648:ultrastructure
D017931:DNA Primers; D011994:Recombinant Proteins