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health hazards to timber and forestry workers from the douglas fir tussock moth.a heavy infestation of the tussock moth caterpillar (orgyia pseudotsugata mcdonnough) in forested areas of oregon was associated with itching of the skin and eyes, nasal discharge, cough, and, at times, respiratory difficulty, personal interviews and inspection of forty-one occupationally exposed persons were supplemented by a questionnaire administered to 428 individuals, composing three groups at various degrees of risk and a control group. there was a cause and effect relationship between the ...1977143915
douglas-fir tussock moth: sex pheromone identification and synthesis.the sex pheromone of the douglass-fir tussock moth orgyia pseudotsugata (mcdunnough) has been isolated and identified as (z)-6-heneicosen-11-one. this compound and its e isomer have been synthesized and are highly potent in laboratory bioassays and field trials.19751114342
baculovirus gp64 envelope glycoprotein is sufficient to mediate ph-dependent membrane fusion.the baculovirus gp64 envelope glycoprotein is a major component of the envelope of the budded virus (bv) and is involved in bv entry into the host cell by endocytosis. to determine whether gp64 alone was sufficient to mediate membrane fusion, the orgyia pseudotsugata multicapsid nuclear polyhedrosis virus gp64 protein was transiently expressed in uninfected insect cells. cells expressing the baculovirus gp64 protein were examined for membrane fusion activity by using a syncytium formation assay ...19921404622
a synthetic early promoter from a baculovirus: roles of the tata box and conserved start site cagt sequence in basal levels of transcription.many baculovirus early genes and insect genes transcribed by rna polymerase ii have a conserved transcription start site sequence (cagt) located downstream of a consensus tata box. to examine the functions and interactions of these two motifs in initiating accurately positioned basal transcription, a 43-nt synthetic promoter was synthesized from the tata box and start site sequences of the gp64 early promoter from the orgyia pseudotsugata multicapsid nuclear polyhedrosis virus (opmnpv). the synt ...19921519359
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.the 67.2- to 68.5-m.u. region of autographa californica nuclear polyhedrosis virus was sequenced. a large open reading frame (orf) was identified in the clockwise direction on the circular genome map which could potentially encode an 80-kda polypeptide. analysis of the predicted amino acid sequence of this orf indicated that it was a homologue of the p87 capsid-associated gene of orgyia pseudotsugata mnpv with an overall amino acid similarity of 34.3%. a late transcript of 2.1 kb was mapped to t ...19921529529
the anticarsia gemmatalis nuclear polyhedrosis virus polyhedrin gene region: sequence analysis, gene product and structural comparisons.the genomic region of the anticarsia gemmatalis multiple nucleocapsid nuclear polyhedrosis virus (agmnpv) strain 2d encoding the polyhedrin gene was cloned and mapped, and a 2085 bp sphi-psti fragment containing the gene was sequenced. the polyhedrin polypeptide of the parental isolate agmnpv was manually sequenced, and the amino acid sequence obtained agreed with that deduced from the dna coding region sequence. agmnpv and orgyia pseudotsugata mnpv (opmnpv) are similar in terms of promoter stru ...19921588315
nucleotide sequence of the p39-capsid gene region of the lymantria dispar nuclear polyhedrosis virus.a 1.85 kb region, containing an open reading frame (orf) homologous to the baculovirus p39-capsid gene, was sequenced from the lymantria dispar multicapsid nuclear polyhedrosis virus (ldmnpv) genome. analysis of the p39-capsid gene demonstrated that it was 39% and 47% identical in amino acid sequence with the homologous genes in the autographa californica and orgyia pseudotsugata mnpvs, respectively. two late promoter elements located upstream of the p39 gene in the ldmnpv genome are conserved w ...19921607869
the basic dna-binding protein of bombyx mori nuclear polyhedrosis virus: the existence of an additional arginine repeat.the basic dna-binding protein of the bombyx mori nuclear polyhedrosis virus (bmnpv) was purified by hplc and a sequence of 45 amino acids from the n-terminus was determined. there were no detectable modifications such as n-terminal blockage, glycosylation, or phosphorylation. the amino acid sequence showed high homology to the predicted amino acid sequences of the basic proteins of autographa californica npv (acnpv) and orgyia pseudotsugata npv (opnpv) (90 and 76%, respectively), however, the bm ...19911703373
the glycoprotein of thogoto virus (a tick-borne orthomyxo-like virus) is related to the baculovirus glycoprotein gp64.thogoto (tho) virus is a tick-transmitted virus which shares morphological and biochemical characteristics with members of the orthomyxoviridae. the genome of thogoto virus comprises six segments of single-stranded, negative sense rna. the complete nucleotide sequence of the fourth largest rna segment of tho virus has been determined from cdna analyses. this rna segment is 1574 nt long and has a coding capacity for a glycoprotein of 512 amino acids with a predicted molecular weight of 57,550 da. ...19921733105
molecular analysis of the trans-activating ie-2 gene of orgyia pseudotsugata multicapsid nuclear polyhedrosis virus.a second immediate early (ie) regulatory gene of the baculovirus orgyia pseudotsugata multicapsid nuclear polyhedrosis virus (opmnpv) has been identified. the ie-2 gene which is homologous to the ie-n gene of autographa californica mnpv was mapped to the hindiii a fragment of opmnpv between 0.41 to 1.37 map units. the ie-2 gene codes for a predicted protein of 45,640 da and analysis of the amino acid sequence shows that the protein has a highly basic amino terminal domain and a cysteine-rich dom ...19921736546
tandemly repeated sequence at the 3' end of the ie-2 gene of the baculovirus orgyia pseudotsugata multicapsid nuclear polyhedrosis virus is an enhancer element.an enhancer element was identified in the virus orgyia pseudotsugata multicapsid nuclear polyhedrosis virus (opmnpv) that is located adjacent to the 3' end of the ie-2 gene and 5' to an open reading frame that codes for a predicted protein that has 37% homology to the acmnpv pe-38 gene. the opmnpv enhancer (ope) consists of a 66-bp element that is tandemly repeated partially or completely 12 times. the ope sequences were shown to increase gene expression from the autographa californica mnpv dela ...19921736547
identification and characterization of the ie-1 gene of orgyia pseudotsugata multicapsid nuclear polyhedrosis virus.the ie-1 gene of orgyia pseudotsugata multicapsid nuclear polyhedrosis virus (opmnpv) was mapped between 95.7 and 97.1 map units on the viral genome. sequence analysis of the opmnpv ie-1 gene (opie-1) identified an open reading frame that coded for a predicted protein of 560 amino acids with a molecular weight of 64,775. transcriptional analysis of opmnpv-infected lymantria dispar (ld652y) cells identified two rnas homologous to the opie-1 open reading frame that were 1.7 and 1.9 kb in size. the ...19911989381
immunoelectron microscopic examination of orgyia pseudotsugata multicapsid nuclear polyhedrosis virus-infected lymantria dispar cells: time course and localization of major polyhedron-associated proteins.immunoelectron microscopy was employed to examine the temporal expression and localization of two proteins involved in baculovirus polyhedron assembly (polyhedrin and p10) of orgyia pseudotsugata multicapsid nuclear polyhedrosis virus (opmnpv) in infected lymantria dispar cells. in addition, the association of p10 with the polyhedron envelope (pe) protein was studied. the major capsid protein (p39) was also examined to investigate the association of virion structural proteins with polyhedron for ...19911993872
the p6.5 gene region of a nuclear polyhedrosis virus of orgyia pseudotsugata: dna sequence and transcriptional analysis of four late genes.the gene encoding the basic dna-binding protein (p6.5) of the multicapsid nuclear polyhedrosis virus of orgyia pseudotsugata (opmnpv) was localized by southern blot analysis using a cdna probe containing the autographa californica virus (acmnpv) p6.9 gene. the opmnpv p6.5 gene was mapped to the hindiii g fragment at map unit 67. nucleotide sequence and transcriptional analysis of a 3.26 kb region encompassing this area revealed four open reading frames (orfs 1 to 4) oriented in the same directio ...19902179466
a capsid-associated protein of the multicapsid nuclear polyhedrosis virus of orgyia pseudotsugata: genetic location, sequence, transcriptional mapping, and immunocytochemical characterization.two lambda gt11 clones containing overlapping dna inserts encoding portions of a structural protein gene from orgyia pseudotsugata multicapsid nuclear polyhedrosis virus (opmnpv) were identified by their immunoreactivity with polyclonal antisera produced against purified polyhedra-derived virus. sequence analysis of a 3.6-kb region of the baculovirus genome (map units 69.1-71.6) from which the lambda gt11 inserts originated revealed an open reading frame of 1872 nt (624 amino acids) encoding a p ...19902184573
characterization of the infection cycle of the orgyia pseudotsugata multicapsid nuclear polyhedrosis virus in lymantria dispar cells.to characterize the infection cycle of the orgyia pseudotsugata multicapsid nuclear polyhedrosis virus in lymantria dispar cells, the time course of dna synthesis and polyhedron production, and the onset and rate of budded virus production were investigated at three different m.o.i. (5, 10 and 100). in addition, the time course of expression of three proteins (gp64, p39 and polyhedrin) representative of three temporal classes of baculovirus genes was also analysed using western blot analysis. dn ...19902273385
a baculovirus polyhedron envelope protein: immunogold localization in infected cells and mature polyhedra.a polyclonal antiserum against a trpe fusion protein containing the complete open reading frame of the polyhedron envelope (pe) protein from the nuclear polyhedrosis virus of orgyia pseudotsugata (opmnpv) was used for immunogold staining and electron microscopic examination of polyhedra, isolated polyhedron envelopes, and infected insect cells at selected times postinfection. the antiserum specifically stained the peripheral envelope of mature polyhedra and also stained the envelope structure wh ...19902403704
characterization of baculovirus p10 synthesis using monoclonal antibodies.a series of monoclonal antibodies were produced against virion proteins of the multicapsid nuclear polyhedrosis virus of orgyia pseudotsugata (opmnpv). four of these antibodies reacted with a protein of 14 kd on western blots of electrophoretically separated opmnpv virion proteins. these antibodies were used to identify immunoreactive clones from a lambda gt11 expression library of opmnpv dna. by hybridization of insert dna from the lambda gt11 clones to blots of digests of opmnpv genomic dna, a ...19872442889
cytoplasmic polyhedrosis virus classification by electropherotype; validation by serological analyses and agarose gel electrophoresis.serological analyses of several different cytoplasmic polyhedrosis viruses (cpvs), including two type 1 cpvs from bombyx mori, type 1 cpv from dendrolimus spectabilis, type 12 cpv from autographa gamma, type 2 cpv from inachis io, type 5 cpv from orgyia pseudotsugata and type 5 cpv from heliothis armigera, demonstrated a close correlation between the antigenic properties of the polyhedrin or virus particle structural proteins and the genomic dsrna electropherotypes. the dsrnas of these viruses w ...19892499658
nucleotide sequence, transcriptional mapping, and temporal expression of the gene encoding p39, a major structural protein of the multicapsid nuclear polyhedrosis virus of orgyia pseudotsugata.the gene encoding the 39-kda major structural protein (p39) of orgyia pseudotsugata nuclear polyhedrosis virus (opmnpv) was sequenced and transcriptionally mapped, and its expression was examined at various times postinfection. by northern hybridization, primer extension, and s1 nuclease analysis, we identified p39 mrnas of approximately 2600 nt. by primer extension analysis, we identified two major sets of transcripts which initiated around -48 and -96 nt upstream of the translation start codon ...19892644736
a baculovirus polyhedral envelope-associated protein: genetic location, nucleotide sequence, and immunocytochemical characterization.using a polyclonal mouse antiserum produced against purified virions of the multicapsid nuclear polyhedrosis virus of orgyia pseudotsugata (opmnpv), two immunoreactive lambda gtii clones were identified which contained nonoverlapping insert dnas which mapped to a single open reading frame (orf) in the hindiii-m fragment. analysis of nucleotide sequence data indicates that this orf encodes a protein with a mw of 32.4 kda. a trpe-p32 gene fusion containing the entire p32 orf was constructed, and t ...19892646825
nucleotide sequence of the polyhedrin gene of euxoa scandens cytoplasmic polyhedrosis virus (escpv).the double-stranded rna genome of euxoa scandens cytoplasmic polyhedrosis virus (escpv) was reversely transcribed to the double-stranded dna and cloned into pibi30. the complete nucleotide sequence of cloned genome segment 10, which encodes the virus polyhedrin polypeptide, was determined. the escpv polyhedrin gene consists of 881 bp and possesses an open reading frame that codes for a polypeptide of 269 amino acids (mw 30.1k), consistent with an apparent mw of 30k determined by sds-page for pur ...19892650462
location, sequence, transcriptional mapping, and temporal expression of the gp64 envelope glycoprotein gene of the orgyia pseudotsugata multicapsid nuclear polyhedrosis virus.the gene encoding gp64, the envelope glycoprotein of the budded virus (bv) of orgyia pseudotsugata multicapsid nuclear polyhedrosis virus (opmnpv), was mapped to the hindiii-e fragment of of the viral genome and expression of the gp64 gene was examined at various times postinfection. to locate the gp64 gene, a cross-reacting monoclonal antibody (acv5) (a. w. hohmann and p. faulkner, 1983, virology, 125, 432-444) directed against the gp64 protein of the autographa californica multicapsid nuclear ...19892658304
characterization of the genetic organization of the hindiii m region of the multicapsid nuclear polyhedrosis virus of orgyia pseudotsugata reveals major differences among baculoviruses.a region including the 4 kb hindiii m fragment of the multicapsid nuclear polyhedrosis virus (mnpv) of orgyia pseudotsugata (opmnpv) genome was sequenced, transcriptionally mapped, and compared to the homologous region in the mnpv of autographa californica (acmnpv). five open reading frames (orfs) oriented in the same direction were identified and were found to be expressed at late times post-infection. the mrnas from the five orfs were found to coterminate at a single site downstream of orf 5. ...19892661722
the genome of the multicapsid baculovirus of orgyia pseudotsugata: restriction map and analysis of two sets of gc-rich repeated sequences.five cosmids containing inserts that comprise the complete genome of the multicapsid nuclear polyhedrosis virus of orgyia pseudotsugata were mapped with four restriction enzymes (bg/ii, clai, ssti, xhoi). from these cosmid maps, composite maps of the complete genome were constructed for each restriction enzyme. a region containing repeats of the sequence ggc downstream of the polyhedrin gene was used to probe the genome. it cross-hybridized with a region which, upon sequence analysis, was found ...19882838575
location and nucleotide sequence of the orgyia pseudotsugata single nucleocapsid nuclear polyhedrosis virus polyhedrin gene.a restriction endonuclease map was determined for the orgyia pseudotsugata single nucleocapsid nuclear polyhedrosis virus (snpv) genome. the order of the fragments generated by the enzymes bglii, bamhi and xbai was analysed using double digestion of the total genome and digestion of isolated restriction fragments. the location of the polyhedrin gene was then determined using a cloned polyhedrin gene from the o. pseudotsugata multiple nucleocapsid npv (mnpv) as a hybridization probe. a fragment c ...19863011971
p39, a major baculovirus structural protein: immunocytochemical characterization and genetic location.a series of monoclonal antibodies was produced against proteins from polyhedra-derived virions of the multicapsid nuclear polyhedrosis virus of orgyia pseudotsugata (opmnpv). two of these antibodies (214 and 236) reacted with a protein of 39 kda on western blots of electrophoretically separated opmnpv viron proteins derived from both budded and polyhedra-derived virions. this protein appears to be a major component of both bv and pdv. one of the p39 antibodies was used to characterize p39 synthe ...19883059676
nucleotide sequencing and transcriptional mapping of the orgyia pseudotsugata multicapsid nuclear polyhedrosis virus p10 gene.a 32p-labeled cloned dna fragment (acmnpv hindiii-q) containing one of the repeated sequences and a portion of the p10 gene from autographa californica multicapsid nuclear polyhedrosis virus (acmnpv) was used to probe southern blots containing restriction endonuclease digests of orgyia pseudotsugata muticapsid nuclear polyhedrosis virus (opmnpv) dna. a single 3.6-kb fragment, opmnpv hindiii-q, was hybridized. the opmnpv hindiii-q fragment was cloned into puc-18, mapped with restriction endonucle ...19863526709
the nucleotide sequence of the polyhedrin gene region from the multicapsid baculovirus of orgyia pseudotsugata.the nucleotide sequence was determined of a 1.6-kb restriction fragment containing the polyhedrin gene of the multicapsid nuclear polyhedrosis virus of orgyia pseudotsugata (opmnpv). in addition, both the 5' and 3' termini of the polyhedrin mrna were located in the flanking sequences using s1 mapping procedures. 5' leader, 3' flanking, and open reading frame sequences of 50-52, 145-167, and 735 nucleotides, respectively, were determined, for a total mrna length of about 935 nucleotides. a potent ...19863526710
comparison of the p26 gene region of two baculoviruses.a 1.1-kb region of dna containing the p26 gene of the multicapsid nuclear polyhedrosis virus of orgyia pseudotsugata (opmnpv) was sequenced, transcriptionally mapped, and compared to the same region in the mnpv of autographa californica (acmnpv). the mrna start site of the p26 gene occurs about 22 nucleotides downstream from an a/t-rich putative promoter sequence that is highly conserved between acmnpv and opmnpv. the p26 mrna is transcribed through the p26 gene and coterminates with the p10 gen ...19873686831
inactivation of nuclear polyhedrosis virus (baculovirus subgroup a) by monochromatic uv radiation.monochromatic radiation at wavelengths of 290, 300, 310, and 320 nm inactivated occluded nuclear polyhedrosis virus of the douglas-fir tussock moth, orgyia pseudotsugata (mcdunnough). data indicate that all of the wavelengths are capable of causing virus inactivation; much greater fluences are needed for virus inactivation as the wavelength increases.19853888111
conservation of genome organization in two multicapsid nuclear polyhedrosis viruses.the genome of the multicapsid nuclear polyhedrosis virus of orgyia pseudotsugata was mapped by examining overlapping hindiii fragments from cosmid clones which had been constructed from partial hindiii digests of viral dna. five opmnpv cosmid clones containing fragments encompassing the entire opmnpv genome were hydridized to blots of dna from the multicapsid nuclear polyhedrosis virus of autographa californica. the hybridization pattern indicated that the genomes of these viruses are similarly ...19846092694
characterization of a cytoplasmic polyhedrosis virus infecting manduca sexta.a cytoplasmic polyhedrosis virus (cpv) infecting manduca sexta was characterized. the genome consisted of ten equimolar rna segments ranging in estimated molecular mass from 0.7 to 3.84 mdaltons and with a total estimated molecular mass of 21.75 mdaltons . the sizes of the individual rna segments provisionally placed this cpv within the type 8 classification of payne and rivers. the polyhedrin protein had an estimated molecular mass of 28 kdaltons . six virion-associated proteins were observed, ...19846373659
production of polyhedrin monoclonal antibodies for distinguishing two orgyia pseudotsugata baculoviruses.monoclonal antibodies were produced to polyhedrins from orgyia pseudotsugata multicapsid nuclear polyhedrosis virus (opmnpv) and single-capsid nuclear polyhedrosis virus (opsnpv). although the polyhedrins are closely related, antibodies were selected which allowed differentiation between the two viruses. in an indirect enzyme-linked immunosorbent assay, purified opmnpv and opsnpv polyhedrins could be detected by specific monoclonal antibodies at concentrations as low as 2 and 5 ng/ml, respective ...19846391377
identification of a membrane fusion domain and an oligomerization domain in the baculovirus gp64 envelope fusion protein.the baculovirus gp64 envelope fusion protein (gp64 efp) is the major envelope glycoprotein of the budded virion and has been shown to mediate acid-triggered membrane fusion both in virions and when expressed alone in transfected cells. using site-directed mutagenesis and functional assays for oligomerization, transport, and membrane fusion, we localized two functional domains of gp64 efp. to identify a fusion domain in the gp64 efp of the orgyia pseudotsugata multiple nuclear polyhedrosis virus ...19957533858
replication of orgyia pseudotsugata baculovirus dna: lef-2 and ie-1 are essential and ie-2, p34, and op-iap are stimulatory genes.a transient dna replication assay was used to identify genes located within m.u. 90.5-7.0 of the orgyia pseudotsugata multinucleocapsid nuclear polyhedrosis virus (opmnpv) genome that influenced replication of a reporter plasmid containing an opmnpv origin of replication, when cotransfected into uninfected lymantria dispar cells. the viral transactivator ie-1 and a 2.4-kb subclone were found to be essential for replication. the 2.4-kb region was sequenced and open reading frames were identified. ...19957571435
characterization, sequencing and phylogeny of the ecdysteroid udp-glucosyltransferase gene from two distinct nuclear polyhedrosis viruses isolated from choristoneura fumiferana.the ecdysteroid udp-glucosyltransferase (egt) gene isolated from a plaque-purified isolate of choristoneura fumiferana multinucleocapsid nuclear polyhedrosis virus (cfmnpv) was compared to its homologue from a defective mnpv virus (cfdef) present in wild type virus populations infecting the eastern spruce budworm, c. fumiferana. the egt genes were located in the same relative position within the virus genomes and their genomic location and arrangement were similar to that found in autographa cal ...19957595348
identification, sequence, and transcriptional analysis of lef-3, a gene essential for orgyia pseudotsugata baculovirus dna replication.using a transient complementation assay for dna replication of an origin-containing reporter plasmid, a 2.4-kb region (m.u. 48.3-50.1) of the orgyia pseudotsugata multinucleocapsid nuclear polyhedrosis virus (opmnpv) genome was identified that contains a gene essential for opmnpv dna replication. this region was sequenced and open reading frames were identified. replication assays using subclones from this region identified a gene called late expression factor 3 (lef-3), as the essential replica ...19957618274
the baculovirus gp64 envelope fusion protein: synthesis, oligomerization, and processing.the baculovirus gp64 envelope fusion protein (gp64 efp) is a class i integral membrane protein that enters the secretory pathway and is oligomerized and extensively processed during transport to the plasma membrane. the kinetics of gp64 efp biosynthesis, oligomerization, and processing in orgyia pseudotsugata multicapsid nuclear polyhedrosis virus (opmnpv)-infected lymantria dispar cells were examined by pulse label, pulse-chase, and immunoprecipitation experiments. relative rates of gp64 efp sy ...19957778291
overlapping tata-dependent and tata-independent early promoter activities in the baculovirus gp64 envelope fusion protein gene.in previous studies to characterize basal and activated transcription from the early promoter of the gp64 envelope fusion protein (efp) gene of the orgyia pseudotsugata multicapsid nuclear polyhedrosis virus, the tata box was identified as a functional element, essential for basal transcription from a minimal promoter construct. in the current study, we used discrete deletions and multiple point mutations that removed the functional tata box from larger promoter constructs of the gp64 efp gene t ...19957853477
identification of essential trans-acting regions required for dna replication of the orgyia pseudotsugata multinucleocapsid nuclear polyhedrosis virus: lef-1 is an essential replication gene.a transient replication assay for the identification of baculovirus genes that are essential for replication of an origin-containing reporter plasmid was established for the orgyia pseudotsugata multinucleocapsid nuclear polyhedrosis virus (opmnpv). using a replication origin located on the opmnpv hindiii-n fragment, we identified a subset of cosmids and plasmids from an opmnpv cosmid library that was able to supply all the essential trans-acting factors and support replication of the origin-con ...19957886946
identification and characterization of a second putative origin of dna replication in a baculovirus of orgyia pseudotsugata.a 7.5-kb region (96.8-2.5 m.u.), called op5, of the orgyia pseudotsugata multinucleocapsid nuclear polyhedrosis virus (opmnpv) genome that contains an origin of dna replication was characterized. this region replicates several times more efficiently and is unrelated to the previously identified putative origin of replication located on the viral hindiii-n fragment. in contrast to hindiii-n, the origin on op5 contains repeated sequences with limited sequence identity to the homologous regions fro ...19957886962
nucleotide sequence of the polyhedrin gene region of helicoverpa zea single nucleocapsid nuclear polyhedrosis virus: placement of the virus in lepidopteran nuclear polyhedrosis virus group ii.the polyhedrin gene (polh) of helicoverpa zea single nucleocapsid nuclear polyhedrosis virus (hzsnpv) was identified and shown by sequence analysis of the ecori i genomic fragment to encode a 246 amino acid polypeptide that has greater than 80% sequence identity to known polyhedrins. it is preceded by an at-rich region containing the conserved late promoter motif taag, which was identified as a transcription start point. downstream of polh there were several similarities in genome arrangement to ...19947964631
analysis of the c-polyhedrin genes from different geographical isolates of a type 5 cytoplasmic polyhedrosis virus.the c-polyhedrin genes of two different geographic isolates of a type 5 cytoplasmic polyhedrosis virus (cpv) were cloned. a cpv infecting orgyia pseudotsugata (opcpv), isolated in the pacific northwest of the u.s.a., and a cpv infecting heliothis armigera, isolated in south africa, were studied. both genes were found to be 883 nucleotides in length and encoded a predicted protein of 246 residues (m(r) of 28,890). comparison of the nucleotide sequences of these two viruses with another type 5 geo ...19948046399
an apoptosis-inhibiting gene from a nuclear polyhedrosis virus encoding a polypeptide with cys/his sequence motifs.two different baculovirus genes are known to be able to block apoptosis triggered upon infection of spodoptera frugiperda cells with p35 mutants of the insect baculovirus autographa californica nuclear polyhedrosis virus (acmnpv):p35 (p35-encoding gene) of acmnpv (r. j. clem, m. fechheimer, and l. k. miller, science 254:1388-1390, 1991) and iap (inhibitor of apoptosis gene) of cydia pomonella granulosis virus (cpgv) (n. e. crook, r. j. clem, and l. k. miller, j. virol. 67:2168-2174, 1993). using ...19948139034
identification of genes encoding late expression factors located between 56.0 and 65.4 map units of the autographa californica nuclear polyhedrosis virus genome.using a previously developed method that allows the identification of autographa californica nuclear polyhedrosis virus (acmnpv) genes which stimulate transient expression from a late and a very late viral promoter [passarelli and miller, j. virol., 67, 2149-2158 (1993)], we have identified three genes between 56.0 and 65.4 map units of the acmnpv genome involved in expression from a late and a very late promoter but not from an early viral promoter. one gene, p143, was previously shown to be es ...19938249293
identification and characterization of a putative origin of dna replication in the genome of a baculovirus pathogenic for orgyia pseudotsugata.a four-kilobase (kb) region (hindiii-n, map units 7.0-11.3) of the orgyia pseudotsugata multinucleocapsid nuclear polyhedrosis virus (opmnpv) genome was found to contain sequences that conferred upon plasmids the ability to undergo infection-dependent replication. the plasmid dna appeared to be replicated to form high molecular weight multimers. plasmids with deletions of up to 1.8 kb from either end of the hindiii-n region were replication competent. however, a discrete sequence, contained with ...19938249294
a baculovirus gp64 early promoter is activated by host transcription factor binding to cacgtg and gata elements.the early promoter of the orgyia pseudotsugata multicapsid nuclear polyhedrosis virus gp64 gene is active when transfected into several insect cell lines and does not require viral gene products for transcription in uninfected cells. because previous studies have shown that the gp64 early promoter is activated above basal levels in uninfected cells, host transcription factors are likely to play a role in gp64 activation at early times postinfection. by using nuclear extracts from uninfected sf9 ...19948289385
a 25-kda protein is associated with the envelopes of occluded baculovirus virions.antiserum produced against preoccluded virions of the orgyia pseudotsugata multinucleocapsid nuclear polyhedrosis virus (opmnpv) was used to screen an opmnpv lambda gt11 expression library. one of the immunoreactive clones contained an insert that hybridized to a portion of the opmnpv hindiii-p fragment. a 2-kb region of hindiii-p was sequenced and found to contain three open reading frames, each preceded by a late promoter element. the insert from the lambda gt11 clone was derived from the thir ...19938337828
characterization of an early gene coding for a highly basic 8.9k protein from the orgyia pseudotsugata multicapsid nuclear polyhedrosis virus.a new gene of the baculovirus orgyia pseudotsugata multicapsid nuclear polyhedrosis virus (opmnpv) has been identified that encodes a highly basic 8.9k protein. the gene called p8.9 is expressed as a 0.5 kb transcript by 1 h post-infection and initiates at an early gene motif. the promoter of the 0.5 kb transcript has two upstream elements, repeats i and ii, which are similar to motifs previously characterized in the opmnpv ie-2 gene and the autographa californica nuclear polyhedrosis virus ie-n ...19938345350
nucleotide sequence of the buzura suppressaria single nucleocapsid nuclear polyhedrosis virus polyhedrin gene.a portion of the genome of the buzura suppressaria (lepidoptera) single nucleocapsid nuclear polyhedrosis virus (bssnpv) containing the polyhedrin gene was sequenced. an open reading frame of 738 nucleotides encoded a protein of 246 amino acids and represented the polyhedrin gene. a conserved taag motif, associated with transcriptional start sites in other polyhedrin genes, was identified 51 nucleotides upstream of the bssnpv polyhedrin gene. a putative polyadenylation signal, aataaa, was found ...19938345353
alternative transcriptional initiation as a novel mechanism for regulating expression of a baculovirus trans activator.in this report, we show that the orgyia pseudotsugata nuclear polyhedrosis virus p34 gene, which is homologous to the autographa californica nuclear polyhedrosis virus pe-38 gene, is a trans activator. the predicted p34 protein contains a number of motifs that are similar to those found in other eukaryotic transcriptional trans activators, including a putative zinc finger dna-binding domain, a glutamine-rich domain, and a leucine zipper. northern (rna) blot analysis showed that the p34 gene is e ...19938371344
analysis of the orgyia pseudotsugata multicapsid nuclear polyhedrosis virus trans-activators ie-1 and ie-2 using monoclonal antibodies.we have produced monoclonal antibodies against two orgyia pseudotsugata multicapsid nuclear polyhedrosis virus (opmnpv) transcriptional trans-activators, ie-1 and ie-2. temporal analysis of ie-1 and ie-2 proteins have shown that ie-1 continues to increase in steady-state levels from 0 to 120 h post-infection, whereas ie-2 declines by 36 h post-infection. at least five different electrophoretic forms of ie-1 are present in opmnpv-infected ld652y cells of which three appear to be from the non-spli ...19938376961
a 37-kilodalton glycoprotein from a baculovirus of orgyia pseudotsugata is localized to cytoplasmic inclusion bodies.the gene encoding a 37-kda glycoprotein (gp37) of orgyia pseudotsugata multinucleocapsid nuclear polyhedrosis virus (opmnpv) was located and sequenced. gp37 of opmnpv was found to have 62 and 37% amino acid sequence identity with gp37 of autographa californica multinucleocapsid nuclear polyhedrosis virus (acmnpv) and with a protein reported to be a component of occlusion bodies from choristoneura biennis entomopoxvirus, respectively. the mrna start site of the opmnpv gp37 gene was mapped within ...19938380088
nucleotide sequence of the ubiquitin-39k gene region from the orgyia pseudotsugata multinucleocapsid nuclear polyhedrosis virus genome.the region of the orgyia pseudotsugata multinucleocapsid nuclear polyhedrosis virus (opmnpv) genome containing the ubiquitin and the nuclear matrix-associated protein (39k) genes was sequenced. the first 77 amino acids of the opmnpv ubiquitin open reading frame (orf) showed 84.4% and 79% amino acid sequence identity to autographa californica multinucleocapsid nuclear polyhedrosis virus (acmnpv) and the spodoptera frugiperda insect ubiquitin, respectively. the predicted opmnpv ubiquitin protein c ...19938389803
immunocytochemical characterization of p24, a baculovirus capsid-associated protein.an open reading frame (orf 1) located upstream of the polyhedron envelope protein gene in the orgyia pseudotsugata multinucleocapsid nuclear polyhedrosis virus (opmnpv) genome was cloned in-frame into a trpe bacterial expression vector. the fusion protein produced by this construct was used for the preparation of a monospecific antiserum. western blot analysis of extracts from opmnpv-infected lymantria dispar cells and autographa californica npv (acmnpv)-infected spodoptera frugiperda cells dete ...19938423444
nucleotide sequence and transcriptional analysis of the p10 gene of spodoptera exigua nuclear polyhedrosis virus.the p10 gene of spodoptera exigua multiple nuclear polyhedrosis virus (semnpv) was localized on the xbai fragment h (5.1 kb) of the physical map of the viral genome. the coding sequence of the semnpv p10 gene is 264 nucleotides (nt) long corresponding to a predicted protein of 88 amino acids with an mhf of 9607. the semnpv p10 protein showed only limited amino acid identity (39% and 26%, respectively) to those of orgyia pseudotsugata mnpv (opmnpv) and autographa californica mnpv (acmnpv) and thu ...19938509757
analysis of the role of 5' promoter elements and 3' flanking sequences on the expression of a baculovirus polyhedron envelope protein gene.the polyhedron envelope protein gene of the orgyia pseudotsugata multinucleocapsid nuclear polyhedrosis virus (opmnpv) is the third in a series of five open reading frames (orfs 1-5) oriented in the same direction. individual mrnas initiate at conserved late gene promoter/mrna start site (a/gtaag) sequences located upstream of each orf and the mrnas co-terminate after the fifth orf. to examine the influence of transcription from upstream promoter elements and the presence of an extensive 3' flan ...19938517021
a baculovirus encoded 16-kda glycoprotein localizes near the nuclear membrane of infected cells.an open reading frame (orf 2) located upstream of the polyhedron envelope protein gene of orgyia pseudotsugata multicapsid nuclear polyhedrosis virus (opmnpv) was cloned in frame into a trpe expression vector. the fusion protein produced by this construct was used for the production of a monospecific antiserum. western blot analysis of opmnpv-infected lymantria dispar cells detected a 16-kda protein at 24 hr postinfection. the 16-kda protein was determined to be n-glycosylated by tunicamycin tre ...19938517029
the dna polymerase and helicase genes of a baculovirus of orgyia pseudosugata.regions of the genome of the orgyia pseudotsugata multinucleocapsid nuclear polyhedrosis virus (opmnpv) containing the dna polymerase and helicase genes were sequenced. the dna polymerase and helicase genes encode predicted proteins of 985 (112.6 kda) and 1223 (140.5 kda) amino acids and exhibited 63 % and 59 % amino acid identity, respectively, with their homologues in the autographa californica mnpv (acmnpv). the influence of sequence variation between the opmnpv and acmnpv dna polymerase and ...19968609478
characterization of a highly conserved baculovirus structural protein that is specific for occlusion-derived virions.a highly conserved baculovirus late gene called odvp-6e was shown to be a structural protein that is specific for occlusion-derived virus (odv) envelopes. the complete sequence of this gene is presented for both orgyia pseudotsugata nuclear polyhedrosis virus (opmnpv) and cydia pomonella granulosis virus (cpgv). the predicted sizes of the opmnpv and cpgv odvp-6e are 40, 241, and 38,655 respectively. the opmnpv odvp-6e gene was transcriptionally mapped and was shown to initiate from a consensus l ...19968615018
identification of baculovirus gene that promotes autographa californica nuclear polyhedrosis virus replication in a nonpermissive insect cell line.a gene that promotes autographa californica m nuclear polyhedrosis virus (acmnpv) replication in iplb-ld652y cells, a cell line that is nonpermissive for acmnpv, was identified in lymantria dispar m nuclear polyhedrosis virus (ldmnpv). cotransfection of acmnpv dna and a plasmid carrying the ldmnpv gene into iplb-ld652y cells results in acmnpv replication. the gene maps between 43.3 and 43.8 map units on the 162-kbp genome of ldmnpv. it comprises a 218-codon open reading frame and encodes a polyp ...19968642646
cloning and expression of apoptosis inhibitory protein homologs that function to inhibit apoptosis and/or bind tumor necrosis factor receptor-associated factors.baculovirus inhibitors of apoptosis (iaps) act in insect cells to prevent cell death. here we describe three mammalian homologs of iap, miha, mihb, and mihc, and a drosophila iap homolog, diha. each protein bears three baculovirus iap repeats and an n-terminal ring finger motif. apoptosis mediated by interleukin 1beta converting enzyme (ice), which can be inhibited by orgyia pseudotsugata nuclear polyhedrosis virus iap (opiap) and cowpox virus crma, was also inhibited by miha and mihb. as mihb a ...19968643514
the gp64 envelope fusion protein is an essential baculovirus protein required for cell-to-cell transmission of infection.to demonstrate the essential nature of the baculovirus gp64 envelope fusion protein (gp64 efp) and to further examine the role of this protein in infection, we inactivated the gp64 efp gene of autographa californica multicapsid nuclear polyhedrosis virus (acmnpv) and examined the biological properties of this virus in vivo. to provide gp64 efp during construction of the recombinant gp64 efp-null acmnpv baculovirus, we first generated a stably transfected insect cell line (sfpop64-6) that constit ...19968676487
sequence and transcriptional analysis of the ubiquitin gene cluster in the genome of spodoptera exigua nucleopolyhedrovirus.the nucleotide sequence of a 1200 bp dna fragment of spodoptera exigua nucleopolyhedrovirus (semnpv) was determined. this sequence contained a cluster of two open reding frames (orfs), one coding for a viral ubiquitin (v-ubi) and another with homology to orf2 of autographa californica (ac) mnpv and orgyia pseudotsugata (op) mnpv. the vubi orf is 240 nucleotides (nt) long, potentially encoding a protein of 80 amino acids with a predicted molecular mass of 9.4 kda. the amino acid sequence of the v ...19968811032
geographic distribution of cytoplasmic polyhedrosis virus infection in douglas-fir tussock moth larvae, orgyia pseudotsugata, in british columbiawe have examined the geographic distribution and prevalence of insect-specific viral infections in douglas-fir tussock moth populations in british columbia. nucleic acids extracted from field collected egg masses from 10 different locations in british columbia were subjected to agarose gel electrophoresis and southern blot analysis. electrophoresis of nucleic acids after digestion with rnase a at high and low ionic strengths showed that the extra bands detected in agarose gels were double-strand ...19968812604
inhibition of interleukin 1 beta-converting enzyme-mediated apoptosis of mammalian cells by baculovirus iap.apoptosis can be a potent weapon against viral infection and consequently has selected for viruses carrying antiapoptosis genes. two baculovirus proteins, iap and p35, can prevent insect cells from dying in response to infection. p35, which interferes with members of the ced-3 family of cysteine proteases, can also function in mammalian cells. we investigated the ability of iap from orgyia pseudotsugata nuclear polyhedrosis virus to prevent death of mammalian cells. iap was transiently expressed ...19968943013
effects of viral inhibitors of apoptosis in models of mammalian cell death.apoptotic cell death is used as a defence against infection by viruses. to counter this protective mechanism, some viruses carry genes whose products can inhibit progression of the apoptotic process in the host cell. as it is clear that the core cell death mechanisms have been conserved through evolution, viral genes from various sources can be used to unravel these mechanisms in mammalian cells. we have produced transgenic mice that express the cowpox gene crma in their t cell compartment, and ...19968950470
nucleotide sequence, insertional mutagenesis, and transcriptional mapping of a conserved region of the baculovirus autographa californica nuclear polyhedrosis virus (map unit 64.8-66.9).the nucleotide sequence of a 2773-bp region of the autographa californica nuclear polyhedrosis virus hr3 variant (acmnpv) (map unit 64.4-68.2) has been determined. this region lies between the previously analyzed p6.9 and p80 genes. three open reading frames were contained within this region, potentially coding for proteins of 45, 40, and 12 kda. the 5' ends of transcripts capable of coding for the 45- and 12-kda proteins map to consensus baculovirus late transcription start sites (ataag), while ...19968989865
association of a baculovirus-encoded protein with the capsid basal region.an open reading frame homologous to acmnpv orf9 (orf1629) was characterized in the orgyia pseudotsugata multinucleocapsid nuclear polyhedrosis virus (opmnpv). sequence analysis indicated that the opmnpv homolog (called orf2) encoded a protein predicted to contain 545 amino acids with a molecular weight of 61 kda. the first 80 amino acids did not have a counterpart in the acmnpv homolog. the remainder of the orf was poorly conserved with 29% amino acid identity overall with the acmnpv orf. howeve ...19979007067
detection and identification of multiple baculoviruses using the polymerase chain reaction (pcr) and restriction endonuclease analysis.a technique using the polymerase chain reaction (pcr) and restriction analysis was developed for the simultaneous detection of eight baculoviruses. the baculoviruses detected by this technique were autographa californica multiple-embedded nuclear polyhedrosis virus (mnpv). anticarsia gemmatalis mnpv, bombyx mori mnpv, orgyia pseudotsugata mnpv. spodoptera frugiperda mnpv, s. exigua mnpv, anagrapha falcifera mnpv, heliothis zea single-embedded nuclear polyhedrosis virus (snpv). a highly conserved ...19979015292
characterization of a unique opmnpv-specific early gene not required for viral infection in tissue culture.opep-2 is an orgyia pseudotsugata multicapsid nucleopolyhedrovirus (opmnpv) early gene in the ie1-ie2 gene region for which there is no homolog in either the archetype virus, autographa californica mnpv, or bombyx mori npv. opep-2 is transcribed immediately upon infection as three mrnas which initiate from a early gene motif (tata-n27-cagt). the expression of multiple transcripts at very early times postinfection has only been previously described for the baculovirus early gene ie1, which produc ...19979018144
the sequence of the orgyia pseudotsugata multinucleocapsid nuclear polyhedrosis virus genome.the nucleotide sequence of the orgyia pseudotsugata multinucleocapsid nuclear polyhedrosis virus (opmnpv) genome was completed and analyzed. it is composed of 131,990 bases with a g + c content of 55% and contains 152 putative genes of 150 nucleotides or greater. major differences in gene content and arrangement between opmnpv and the autographa californica mnpv were found. these include the presence in opmnpv of three complete iap gene homologs, two conotoxin gene homologs, two protein tyrosine ...19979126251
baculovirus immediate-early promoter-mediated expression of the zeocin resistance gene for use as a dominant selectable marker in dipteran and lepidopteran insect cell lines.the antibiotic zeocin, a derivative of phleomycin, was evaluated for use as a selection system in both dipteran and lepidopteran insect cell lines. growth of drosophila cell lines, kc1 and sl2, was inhibited at zeocin concentrations of 50 and 75 microg/ml, respectively, while the spodoptera cell line, sf9, was inhibited at a concentration of 250 microg/ml zeocin. the mammalian cytomegalovirus (cmv) and simian virus 40 (sv40) early promoters did not function in these insect cell lines. several ba ...19979133590
baculovirus inhibitor of apoptosis functions at or upstream of the apoptotic suppressor p35 to prevent programmed cell death.members of the inhibitor of apoptosis (iap) gene family prevent programmed cell death induced by multiple signals in diverse organisms, suggesting that they act at a conserved step in the apoptotic pathway. to investigate the molecular mechanism of iap function, we expressed epitope-tagged op-iap, the prototype viral iap from orgyia pseudotsugata nuclear polyhedrosis virus, by using novel baculovirus recombinants and stably transfected insect cell lines. epitope-tagged op-iap blocked both virus- ...19979151843
late promoter selection in the baculovirus gp64 envelope fusion protein gene.the upstream promoter region of the autographa californica multicapsid nuclear polyhedrosis virus (acmnpv) gp64 gene contains five copies of taag, the conserved sequence found at the transcriptional initiation sites of almost all baculovirus late genes. in acmnpv-infected sf9 cells, late transcription initiation is detected from only two upstream taag sites and not from three downstream taag sites. to examine several models for preferential taag site utilization, we constructed a series of recom ...19979168879
characterization of p91, a protein associated with virions of an orgyia pseudotsugata baculovirus.polyclonal antiserum produced against preoccluded virions from the orgyia pseudotsugata multinucleocapsid nuclear polyhedrosis virus (opmnpv) was used to screen an opmnpv lambda gt11 expression library. the insert from one of the immunoreactive phage isolates hybridized to opmnpv orf86 (p91), a 2460-bp (819 amino acids) open reading frame that encodes a predicted protein of 91 kda. antibodies generated against a maltose binding protein-p91 fusion detected a band of approximately 91 kda on wester ...19979201231
the p10 gene of spodoptera littoralis nucleopolyhedrovirus: nucleotide sequence, transcriptional analysis and unique gene organization in the p10 locus.the p10 gene of the spodoptera littoralis (spli) multicapsid nucleopolyhedrovirus (mnpv) was identified. with a coding sequence of 315 nucleotides (nt), corresponding to a protein of 104 amino acids, the splimnpv p10 gene is the longest p10 gene known. this gene codes for a putative protein with an mr of 11130 and was found to be most closely related to the spodoptera exigua (se) mnpv p10 (49.4% amino acid identity) and most distant from the autographa californica (ac) mnpv p10 (20.0% amino acid ...19979267016
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 ...19979349489
a series of broad host range shuttle vectors for constitutive and inducible expression of heterologous proteins in insect cell lines.a series of shuttle vectors have been constructed that allow expression of heterologous proteins in either dipteran or lepidopteran insect cell lines. constitutive expression in a broad range of host cells is mediated by the orgyia pseudotsugata multicapsid nucleopolyhedrosis virus (opmnpv) immediate-early 2 (ie2) promoter. alternatively, if inducible expression is required, for example to express cytotoxic proteins, a vector has been constructed that uses the drosophila metallothionein (mtn) pr ...19989511767
molecular characterization and expression of the trichoplusia ni granulovirus helicase gene.a putative dna helicase gene from the granulovirus of trichoplusia ni (tngv) was cloned, sequenced, and compared with the corresponding gene of several multinucleocapsid nucleopolyhedroviruses (mnpvs) including those from autographa californica (acmnpv), orgyia pseudotsugata (opmnpv), bombyx mori (bmnpv), and spodoptera exigua (semnpv). the tngv helicase gene (p137) encoded a helicase of 1158 amino acids with a predicted mass of 137 kda. comparison of p137 with acmnpv p143 revealed 44.5% identit ...19989603347
identification and characterization of the spodoptera littoralis nucleopolyhedrovirus type b lef-3 gene.we have identified a gene from the spodoptera littoralis nucleopolyhedrovirus type b (splinpv-b) with several characteristics that suggest that it is homologous to the lef-3 genes of the autographa californica and orgyia pseudotsugata npvs (acmnpv and opmnpv, respectively). the splinpv-b lef-3 gene was mapped between 43.6 and 45.5 map units of the splinpv-b genome. northern blot analysis showed that splinpv-b lef-3 was expressed as a 1.6 kb transcript at 5 h post infection (p.i.), reached high l ...19989638145
a highly conserved genomic region in baculoviruses: sequence analysis of an 11.3 kbp dna fragment (46.5-55.1 m.u.) of the spodoptera exigua multicapsid nucleopolyhedrovirus.a dna fragment of 11.3 kilobase pairs (kbp) in size of the baculovirus spodoptera exigua multicapsid nucleopolyhedrovirus (semnpv) genome (46.5 to 55.1 m.u.) was completely sequenced. analysis of the sequence revealed eleven potential open reading frames (orf). ten of these orfs showed significant amino acid identity to autographa californica mnpv (acmnpv) and orgyia pseudotsugata mnpv (opmnpv) genes p6.9, lef5, 38k, p19, p143, p25, p18, vp33, lef4, and vp39. one orf (xc12) has no homolog in oth ...19989725671
new cell lines from heliothis virescens: characterization and susceptibility to baculovirusesnew cell lines from embryos of heliothis virescens were recently developed. six primary cultures were initiated in june 1995. from these initial cultures, two produced sufficient cell growth to allow subcultivation and eventually led to the establishment of seven cell strains, three of which are maintained at low temperatures (17 degreesc). the strains were compared with a previously established cell line from h. virescens by isozyme analysis and shown to be from the same species. all the strain ...19989784351
a mutational analysis of the baculovirus inhibitor of apoptosis op-iap.a family of antiapoptotic regulators known as inhibitors of apoptosis (iaps) was initially identified and functionally described in baculoviruses, and iap homologues are now known in insects, birds, and mammals. baculovirus and drosophila iaps inhibit apoptosis induced by drosophila proapoptotic proteins reaper, hid, and grim and physically interact with them through their baculovirus iap repeat (bir) region. here we examined the functional importance of bir and ring finger motifs of orgyia pseu ...19989852042
characterization of the acidic domain of the ie1 regulatory protein from orgyia pseudotsugata multicapsid nucleopolyhedrovirus.this study presents a detailed analysis of the acidic n-terminal region of the orgyia pseudotsugata multicapsid nucleopolyhedrovirus (opmnpv) transactivator ie1. the n-terminal region of ie1 is rich in acidic amino acids and has been hypothesized to be an acidic activation domain. removal of the n-terminal 126 amino acids containing the acidic domain of ie1 resulted in complete loss of transactivation activity, indicating that this region is essential for transactivation. the opmnpv acidic domai ...19989875318
host cell receptor binding by baculovirus gp64 and kinetics of virion entry.gp64 is the major envelope glycoprotein from budded virions of the baculoviruses autographa californica multicapsid nucleopolyhedrovirus (acmnpv) and orgyia pseudotsugata multicapsid nucleopolyhedrovirus (opmnpv). to examine the potential role of gp64 as a viral attachment protein in host cell receptor binding, we generated, overexpressed, and characterized a soluble form of the opmnpv gp64 protein, gp64solop. assays for trimerization, sensitivity to proteinase k, and reduction by dithiothreitol ...199910366584
molecular characterization of genes in the gp41 region of baculoviruses and phylogenetic analysis based upon gp41 and polyhedrin genes.a newly sequenced anticarsia gemmatalis multiple nucleopolyhedrovirus (agmnpv) gp41 gene was used to reconstruct the phylogeny for gp41 by comparison with autographa californica mnpv, bombyx mori mnpv, helicoverpa zea single nucleopolyhedrovirus (snpv), lymantria dispar mnpv, orgyia pseudotsugata mnpv and spodoptera frugiperda mnpv. the 3.5 kb fragment of the agmnpv gp41 region not only contained the gp41 gene but also three other open reading frames that had significant homology with the very l ...199910518714
a baculovirus anti-apoptosis gene homolog of the trichoplusia ni granulovirus.an inhibitor of apoptosis (iap) gene homolog (tn-iap) of the trichoplusia ni granulovirus (tngv) was cloned, sequenced and mapped on the genome of tngv. tn-iap encoded a protein (tn-iap) of 301 amino acids with a predicted molecular mass of 35 kda. the tn-iap contained the two sequence motifs, birs and ring finger, characteristic of iap proteins, and shared identities of 21-27% and similarities of 28-53% with iap proteins of cydia pomonella gv (cp-iap), orgyia pseudotsugata multinucleocapsid nuc ...199910541013
sequence and organization of the spodoptera exigua multicapsid nucleopolyhedrovirus genome.the nucleotide sequence of the dna genome of spodoptera exigua multicapsid nucleopolyhedrovirus (semnpv), a group ii npv, was determined and analysed. the genome contains 135611 bp and has a g+c content of 44 mol%. computer-assisted analysis revealed 139 orfs of 150 nucleotides or larger; 103 have homologues in autographa californica mnpv (acmnpv) and a further 16 have homologues in other baculoviruses. twenty orfs are unique to semnpv. major differences in semnpv gene content and arrangement we ...199910567663
filamentous actin is required for lepidopteran nucleopolyhedrovirus progeny production.autographa californica m nucleopolyhedrovirus (acmnpv) is the prototypical member of the nucleopolyhedrosis: genus of the baculoviridae:, a family of large, double-stranded dna viruses that are highly diverse. nucleocapsid morphogenesis of acmnpv and others in the nucleopolyhedrovirus: genus takes place within the nuclei of infected host cells. previously, we showed that filamentous actin (f-actin) is essential for this process to occur in acmnpv-infected cells, an unprecedented finding for a dn ...200010859396
a novel baculovirus envelope fusion protein with a proprotein convertase cleavage site.the entry mechanism of spodoptera exigua multicapsid nucleopolyhedrovirus (semnpv), a group ii npv, in cultured cells was examined. semnpv budded virus (bv) enters by endocytosis as do the bvs of the group i npvs, autographa californica (ac) mnpv and orgyia pseudotsugata (op) mnpv. in group i npvs, upon infection acidification of the endosome triggers fusion of the viral and endosomal membrane, which is mediated by the bv envelope glycoprotein gp64. however, the semnpv genome lacks a homolog of ...200011017785
the sequence of the helicoverpa armigera single nucleocapsid nucleopolyhedrovirus genome.the nucleotide sequence of the helicoverpa armigera single-nucleocapsid nucleopolyhedrovirus (hasnpv) dna genome was determined and analysed. the circular genome encompasses 131,403 bp, has a g+c content of 39.1 mol% and contains five homologous regions with a unique pattern of repeats. computer-assisted analysis revealed 135 putative orfs of 150 nt or larger; 100 orfs have homologues in autographa californica multicapsid npv (acmnpv) and a further 15 orfs have homologues in other baculoviruses ...200111125177
production of functional antibodies generated in a nonlytic insect cell expression system.a monoclonal antibody directed against the type 2 adenovirus (ad2) penton base protein was cloned and expressed in spodoptera frugiperda (sf9) cells using a nonlytic vector system. the coding sequences for the immunoglobulin light and heavy chains were placed under the control of the orgyia pseudotsugata multicapsid nucleopolyhedrosis virus immediate-early 2 (opie2) promoter. transfected sf9 cells continuously secreted the antibody which retained the ability to recognize both native and recombin ...200111162396
identification, sequence analysis, and phylogeny of the immediate early gene 1 of the trichoplusia ni single nucleocapsid polyhedrosis virus.substantial research has been conducted on the immediate early i (ie-1) genes from the prototype baculovirus auographa californica multicapsid nuclear polyhedrosis virus (acmnpv) and the orgyia pseudotsugata multicapsid nuclear polyhedrosis virus (opmnpv). in both cases ie-1 gene products have been implicated in transcriptional activation and repression. in this study an ie-1 homolog was identified from trichoplusia ni single nucleocapsid polyhedrosis virus (tnisnpv). nucleotide sequence analysi ...200111556402
measurement of membrane fusion activity from viral membrane fusion proteins based on a fusion-dependent promoter induction system in insect cells.a number of viral membrane fusion proteins can be expressed alone on the surface of host cells, and then triggered to induce cell-to-cell fusion or syncytium formation. although rapid and easily observed, syncytium formation is not easily quantified and differences in fusion activity are not easily distinguished or measured. to address this problem, we developed a rapid and quantitative cell-to-cell fusion system that is useful for comparative analysis and may be suitable for high throughput scr ...200111562545
inhibitor of apoptosis protein from orgyia pseudotsugata nuclear polyhedrosis virus provides a costimulatory signal required for optimal proliferation of developing thymocytes.the inhibitors of apoptosis proteins (iaps) constitute a family of endogenous inhibitors that control apoptosis in the cell by inhibiting caspase processing and activity. iaps are also implicated in cell division, cell cycle regulation, and cancer. to address the role of iaps in thymus development and homeostasis, we generated transgenic mice expressing iap generated from the baculovirus orgyia pseudotsugata nuclear polyhedrosis virus (opiap). developing thymocytes expressing opiap show increase ...200211823509
expression of the human mu opioid receptor in a stable sf9 cell line.the cdna that encodes the human mu opioid receptor (hmor) has been cloned and expressed in spodoptera frugiperda (sf9) cells using a nonlytic vector system. the coding sequence fused to the cleavable glycoprotein signal peptide gp 64, and a c-terminal histidine tag was placed under the transcriptional control of the orgyia pseudotsugata multicapsid nucleopolyhedrosis virus immediate-early 2 (opie2) promoter. transfected cells were selected using zeocin resistance and the receptor was constitutiv ...200211911927
the acidic activation domain of the baculovirus transactivator ie1 contains a virus-specific domain essential for dna replication.ie1 is a potent transcriptional transactivator of the baculovirus orgyia pseudotsugata multiple nucleopolyhedrovirus (opmnpv) and has been shown to be essential for viral dna replication. ie1 contains an acidic activation domain (aad) at the n terminus that is essential for transcriptional transactivation, but its role in viral dna replication is unknown. in this study the role of the ie1 aad in dna replication is investigated. we have determined that deletion of the aad eliminates the ability o ...200211991988
characterization of the p10 gene region of anticarsia gemmatalis nucleopolyhedrovirus.the anticarsia gemmatalis nucleopolyhedrovirus (agmnpv) p10 gene region was cloned, sequenced and the putative p10 gene expression characterized by northern-blot analysis. sequence analysis of the p10 gene region indicated the presence of two complete open reading frames (orfs) of 713 and 281 nucleotides, which codes for polypeptides of 273 and 93 amino acids, with homology to the p26 and p10 proteins of baculoviruses, respectively. two additional partial orfs, coding for partial polypeptides of ...200212086145
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