The gene encoding the capsid protein P82 of the Choristoneura fumiferana multicapsid nucleopolyhedrovirus: sequencing, transcription and characterization by immunoblot analysis.
A gene encoding a capsid-associated viral structural protein has been identified and sequenced in the genome of the Choristoneura fumiferana multicapsid nucleopolyhedrovirus (CfMNPV). The gene has a 1872 nucleotide open reading frame (ORF) encoding 624 amino acids with a predicted molecular mass of 71.4 kDa. Transcription, which appeared to be initiated from a conserved GTAAG motif of baculovirus late genes, was detected at 12 h, reached a maximum at 48 h and declined at 72 h post-infection (p.i.). Part of the ORF was cloned in frame into a prokaryotic expression vector, pMAL-c2, and the fusion protein was used to generate antibodies in rabbits. It was shown, with the aid of the polyclonal antiserum, that this viral protein was detectable at 24 h p.i. in infected cells. The protein appeared as an 82 kDa band in occlusion-derived virus and as an 82 kDa band and a 72 kDa band in budded virus. Amino acid sequence comparisons revealed that this ORF had high homology with the ORF p87 (77% similarity) of Orgyia pseudotsugata (Op) MNPV and the ORF p80 (60% similarity) of Autographa californica (Ac) MNPV. Immunoblots confirmed that the CfMNPV protein had antigenic similarities to the P87 protein of OpMNPV, but not to the P80 of AcMNPV.
Li
X
X
Laboratory for Molecular Virology, Sault Ste. Marie, Ontario, Canada.
Pang
A
A
Lauzon
H A
HA
Sohi
S S
SS
Arif
B M
BM
D000595:Amino Acid Sequence; D000818:Animals; D000914:Antibodies, Viral / Q000276:immunology; D001483:Base Sequence; D015153:Blotting, Western; D002213:Capsid / Q000235:genetics*; D036022:Capsid Proteins; D005814:Genes, Viral; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009036:Moths / Q000821:virology; D017924:Nucleopolyhedroviruses / Q000235:genetics*; D016366:Open Reading Frames; D012333:RNA, Messenger / Q000235:genetics; D012367:RNA, Viral / Q000235:genetics; D015183:Restriction Mapping; D016415:Sequence Alignment; D012995:Solubility; D014158:Transcription, Genetic; D015678:Viral Structural Proteins / Q000235:genetics*
D000914:Antibodies, Viral; D036022:Capsid Proteins; D012333:RNA, Messenger; D012367:RNA, Viral; D015678:Viral Structural Proteins; C076630:Ac-vp80 protein, Autographa californica nucleopolyhedrovirus