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.
ArifB. M.,
KuzioJ.,
FaulknerP.,
DoerflerW.1984; The genome of Choristoneura fumiferana nuclear polyhedrosis virus: molecular cloning and mapping of the Eco RI, Bam HI, Sma I, Xba I and Bgl II restriction sites. Virus Research 1:605–614
BarrettJ. W.,
KrellP. J.,
ArifB. M.1995; Characterization, sequencing and phylogeny of the ecdysteroid UDP-glucosyltransferase gene from two distinct nuclear polyhedrosis viruses isolated from Choristoneura fumiferana
. Journal of General Virology 76:2447–2456
BarrettJ. W.,
LauzonH. A. M.,
MercuriP. S.,
KrellP. J.,
SohiS. S.,
ArifB. M.1996; The putative LEF-1 proteins from two distinct Choristoneura fumiferana multiple nucleopolyhedroviruses share domain homology to eukaryotic primases. Virus Genes 13:229–237
BraunagelS. C.,
EltonD. M.,
MaH.,
SummersM. D.1996a; Identification and analysis of an Autographa californica nuclear polyhedro- sis virus structural protein of the occlusion-derived virus envelope: ODV-E56. Virology 217:97–110
ClarkeE. E.,
TristemM.,
CoryJ. S.,
O’ReillyD. R.1996; Characterization of the ecdysteroid UDP-glucosyltransferase gene from Mamestra brassicae nucleopolyhedrovirus. Journal of General Virology 77:2865–2871
HongT.,
BraunagelS. C.,
SummersM. D.1994; Transcription, translation, and cellular localization of PDV-E66 : a structural protein of the PDV envelope of Autographa californica nuclear polyhedrosis virus. Virology 204:210–222
HuZ. H.,
BroerR.,
WesterlakenJ.,
MartensJ. W. M.,
JinF.,
JehleJ. A.,
WangL. M.,
VlakJ. M.1997; Characterization of the ecdysteroid UDP-glucosyl-transferase gene of a single nucleocapsid nucleopoly- hedrovirus of Buzura suppressaria
. Virus Research 47:91–97
LuA.,
CarstensE. B.1992; Nucleotide sequence and transcriptional analysis of the p80 gene of Autographa californica nuclear polyhedrosis virus : a homologue of the Orgyia pseudotsugata nuclear polyhedrosis virus capsid-associated gene. Virology 190:201–209
MüllerR.,
PearsonM. N.,
RussellR. L. Q.,
RohrmannG. F.1990; A capsid-associated protein of the multicapsid nuclear polyhedrosis virus of Orgyia pseudotsugata : genetic location, sequence, transcriptional mapping, and immunocytochemical characterization. Virology 176:133–144
RussellR. L. Q.,
PearsonM. N.,
RohrmannG. F.1991; Immuno- electron microscopic examination of Orgyia pseudotsugata multicapsid nuclear polyhedrosis virus-infected Lymantria dispar cells : time course and localization of major polyhedron-associated proteins. Journal of General Virology 72:275–283
TheilmannD. A.,
ChantlerJ. K.,
StewartS.,
FlipsenH. T. M.,
VlakJ. M.,
CrookN. E.1996; Characterization of a highly conserved baculovirus structural protein that is specific for occlusion-derived virions. Virology 218:148–158
The gene encoding the capsid protein P82 of the Choristoneura fumiferana multicapsid nucleopolyhedrovirus: sequencing, transcription and characterization by immunoblot analysis