Publications

temperature dependent characteristics of a recombinant infectious hematopoietic necrosis virus glycoprotein produced in insect cells.	a recombinant infectious hematopoietic necrosis virus (ihnv) glycoprotein (g protein) was produced in insect cells using a baculovirus vector (autographa californica nuclear polyhedrosis virus). characteristics of this protein were evaluated in relation to native viral g protein. a full-length (1.6 kb) cdna copy of the glycoprotein gene of ihnv was inserted into the baculovirus vector under control of the polyhedrin promoter. high levels of g protein (approximately 0.5 microgram/1 x 10(5) cells) ...	1999	10349547
generation of a new protein purification matrix by loading ceramic hydroxyapatite with metal ions--demonstration with poly-histidine tagged green fluorescent protein.	the gene encoding the green fluorescent protein (gfp) from the jellyfish aequorea victoria, was inserted under transcriptional control of the polyhedrin promoter of the autographa californica nuclear polyhedrosis virus and expressed in the spodoptera frugiperda insect cell line sf9 during viral infection. the baculovirus transfervector pbluebachisb was used for constructing the recombinant baculovirus, so that the green fluorescent protein could be tagged with a poly-histidine tail. this fusion ...	1999	10361722
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 ...	1999	10366584
mitochondrial dna acts as potential promoter of the baculovirus rna polymerase.	we have examined whether mitochondrial dna could act as target of the rna polymerase encoded by the baculovirus autographa californica multicapsid nuclear polyhedrosis virus, because the baculovirus late promoters and the control region of host mitochondrial dna show a high degree of sequence similarity. in vitro transcription using mitochondrial dna from spodoptera frugiperda cells and nuclear extracts prepared from baculovirus infected cells demonstrates that mitochondrial dna is recognized by ...	1999	10384964
global protein synthesis shutdown in autographa californica nucleopolyhedrovirus-infected ld652y cells is rescued by trna from uninfected cells.	global protein synthesis arrest occurs in autographa californica nucleopolyhedrovirus (acnpv)-infected ld652y cells at late times postinfection (p.i.). a lymantria dispar nucleopolyhedrovirus gene, hrf-1, precludes this protein synthesis arrest. we used in vitro translation assays to characterize the translation defect. cell-free lysates prepared from uninfected ld652y cells, acnpv-infected cells harvested at early times p.i., and cells infected with vachrf-1, a recombinant acnpv bearing hrf-1, ...	1999	10417257
host-specific transcription of nucleopolyhedrovirus gene homologues in productive and abortive anticarsia gemmatalis mnpv infections.	in a previous report, we showed that anticarsia gemmatalis nucleopolyhedrovirus (agmnpv) infections of choristoneura fumiferana ipri-cf-124t and bombyx mori bm-5 cell lines are abortive, whereas a. gemmatalis ufl-ag-286 cells efficiently produce infectious virus and polyhedral inclusion bodies (pibs). in the present study, we explored transcription patterns in these infections using representative temporal classes of autographa californica mnpv (acmnpv) genes. northern analyses were carried out ...	1999	10446647
role of the 3' untranslated region of baculovirus p10 mrna in high-level expression of foreign genes.	the p10 gene of autographa californica nucleopolyhedrovirus has two putative aataaa polyadenylation signals. the downstream signal is used predominantly, as was determined by analysing 3' cdna ends. this downstream motif is followed by a gt-rich sequence, known to be important for efficient polyadenylation in mammalian systems. to analyse the importance of polyadenylation for p10 gene expression, recombinant viruses with altered 3' untranslated regions (utrs) were tested using chloramphenicol ac ...	1999	10466825
the establishment of heliothine cell lines and their susceptibility to two baculoviruses.	a total of eight cell lines were established from helicoverpa armigera (3) and h. punctigera (5) embryos and ovaries. cell lines were established and grown in tc100 and/or tc199-mk containing 10% fetal bovine serum. the serum-free medlium excell 400 was also used, with and without 10% supplemental fetal bovine serum, but failed to generate cell lines from fat bodies, embryos, or ovarian tissues. cell lines consisted of heterogenous cell types ranging from oval to fibroblast-like. this is the fir ...	1999	10475263
comparison of cell line maintenance procedures on insect cells used for producing baculoviruses.	a gypsy moth cell line, iplb-ldeita, maintained under various conditions was tested for susceptibility to and productivity of two baculoviruses, the autographa californica nucleopolyhedrovirus (acmnpv) and lymantria dispar nucleopolyhedrovirus (ldmnpv). the results suggest that cells maintained in serum-containing medium (modified tc100) were more susceptible (on the basis of titers in an endpoint assay) to ldmnpv than cells maintained in a serum-free medium (excell 400). such a difference was n ...	1999	10475270
the establishment of new cell lines from pseudaletia unipuncta with differential responses to baculovirus infection.	six insect cell lines from pseudaletia unipuncta embryos were established and characterized, and their susceptibility to autographa californica multiple nuclear polyhedrosis virus (acmnpv) infection was investigated. these embryonic p. unipuncta cell lines had characteristics distinct from each other in morphology and growth, and showed differential responses to acmnpv infection. among the six cell lines, two were highly susceptible to virus infection. one of these two cell lines, bti-pu-a7s, pr ...	1999	10476920
mutations within the autographa californica nucleopolyhedrovirus fp25k gene decrease the accumulation of odv-e66 and alter its intranuclear transport.	previous reports indicate that mutations within the autographa californica nucleopolyhedrosis virus fp25k gene (open reading frame 61) significantly reduce incorporation of enveloped nucleocapsids into viral occlusions. we report that fp25k is a nucleocapsid protein of both the budded virus (bv) and occluded virus (odv), and we describe the effects of two fp25k mutations (480-1 [n-terminal truncation] and fp-betagal [c-terminal fusion]) on the expression and cellular localization of odv-e66 and ...	1999	10482609
the characterization and phylogenetic relationship of the trichoplusia ni single capsid nuclear polyhedrosis virus polyhedrin gene.	the polyhedrin gene (polh) was identified from the trichoplusia ni (tni) single capsid nuclear polyhedrosis virus (snpv). an ecori fragment containing the truncated polyhedrin gene was detected by hybridization with an acmnpv expression vector probe; the remaining portion of the gene was amplified by reverse pcr. an open reading frame (orf) of 741 nucleotides (nt), encoding a putative protein of 246 amino acids (a.a) with mr 28,780 da was identified. the 5'-noncoding region contained the putativ ...	1999	10499452
purification of autographa californica nucleopolyhedrovirus dna polymerase from infected insect cells.	autographa californica nucleopolyhedrovirus (acmnpv) dna polymerase was purified from virus-infected cells using conventional chromatographic methods. the enzymatic activity of fractions eluting from single-stranded agarose gels was found to exactly coincide with a single polypeptide with an apparent molecular mass of approximately 110,000 da on denaturing polyacrylamide gels stained with coomassie blue. this purification scheme resulted in a 228-fold purification of acmnpv dna polymerase with r ...	1999	10501509
sequence analysis of the xestia c-nigrum granulovirus genome.	the nucleotide sequence of the xestia c-nigrum granulovirus (xcgv) genome was determined and found to comprise 178,733 bases with a g+c content of 40.7%. it contained 181 putative genes of 150 nucleotides or greater that showed minimal overlap. eighty-four of these putative genes, which collectively accounted for 43% of the genome, are homologs of genes previously identified in the autographa californica multinucleocapsid nucleopolyhedrovirus (acmnpv) genome. these homologs showed on average 33% ...	1999	10502508
expression, purification and characterization of peanut (arachis hypogaea) agglutinin (pna) from baculovirus infected insect cells.	peanut (arachis hypogaea) seed lectin, pna is widely used to identify tumor specific antigen (t-antigen), galbeta1-3galnac on the eukaryotic cell surface. the functional amino acid coding region of a cdna clone, pbsh-pn was pcr amplified and cloned downstream of the polyhedrin promoter in the autographa californica nucleopolyhedrovirus (acnpv) based transfer vector pvl1393. co-transfection of spodoptera frugiperda cells (sf9) with the transfer vector, pacpna and acrp6 (a recombinant acnpv having ...	1999	10513900
detection of baculovirus-infected insect cells by flow cytometric side-scatter analyses.	the baculovirus expression vector system (bevs), utilizing the autographa californica nuclear polyhedrosis virus (acnpv), has turned out to be an attractive alternative for high-level expression (<600 mg/l) of recombinant proteins. however, there is a shortage of reliable methods for monitoring the infection process in situations where marker proteins cannot be used.	1999	10520205
characterization of a nucleopolyhedrovirus from the black cutworm, agrotis ipsilon (lepidoptera: noctuidae)	the black cutworm, agrotis ipsilon (hufnagel) (lepidoptera: noctuidae), is a serious localized pest of vegetable and field crops. we have characterized a newly discovered baculovirus, the agrotis ipsilon multicapsid nucleopolyhedrovirus (agipmnpv), that was isolated from a. ipsilon in illinois. restriction enzyme fragment profiles of agipmnpv dna were distinct from those of previously described nucleopolyhedroviruses. electron microscopy of agipmnpv-infected tissues indicated that nucleocapsids ...	1999	10534417
expression of genes coding for animal virus glycoproteins in heterologous systems.	the outermost layers of animal viruses are usually composed of glycoproteins. they are responsible not only for the entrance of viruses into, and release from host cells but also for the initial interaction of a viral particle with immunological defense of the host. it is therefore not surprising that many laboratories devote a lot of effort to study viral glycoproteins at the molecular level. very often such studies are possible only after the introduction of a glycoprotein gene into a heterolo ...	1999	10547034
baculovirus stimulates antiviral effects in mammalian cells.	herein, we report that autographa californica nucleopolyhedrovirus, a member of the baculoviridae family, is capable of stimulating antiviral activity in mammalian cells. baculoviruses are not pathogenic to mammalian cells. nevertheless, live baculovirus is shown here to induce interferons (ifn) from murine and human cell lines and induces in vivo protection of mice from encephalomyocarditis virus infection. monoclonal antibodies specific for the baculovirus envelope gp67 neutralize baculovirus- ...	1999	10559307
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 ...	1999	10567663
the nucleopolyhedroviruses of rachiplusia ou and anagrapha falcifera are isolates of the same virus.	the 7.8 kb ecori-g fragment of rachiplusia ou multicapsid nucleopolyhedrovirus (romnpv), containing the polyhedrin gene, was cloned and sequenced. the sequence of the fragment was 92.3% identical to the sequence of the corresponding region in the autographa californica (ac)mnpv genome. a comparison of the ecori-g sequence with other mnpv sequences revealed that romnpv was most closely related to acmnpv. however, the predicted amino acid sequence of romnpv polyhedrin shared more sequence identity ...	1999	10573177
baculovirus-based genetic screen for antiapoptotic genes identifies a novel iap.	the prototype baculovirus, autographa californica multiple nuclear polyhedrosis virus (acmnpv) expresses p35, a potent anti cell-death gene that promotes the propagation of the virus by blocking host cell apoptosis. infection of insect sf-21 cells with acmnpv lacking p35 induces apoptosis. we have used this pro-apoptotic property of the p35 null virus to screen for genes encoding inhibitors of apoptosis that rescue cells infected with the p35 defective virus. we report here the identification of ...	1999	10593985
development of a novel expression vector system using spodoptera exigua nucleopolyhedrovirus.	to develop a novel spodoptera exigua nucleopolyhedrovirus (senpv) expression vector system, we examined characteristics of the senpv polyhedrin expression in s. exigua cells (se301). while the extracellular virus titer of senpv was 100-fold lower than that of autographa californica nucleopolyhedrovirus (acnpv), the levels of polyhedral inclusion body (pib) formation and polyhedrin expression were higher in senpv. to investigate foreign gene expression under the control of the polyhedrin promoter ...	1999	10597039
mediated pathogenicity of the baculovirus acmnpv by the venom from euplectrus comstockii howard (hymenoptera: eulophidae).	the compatibility of the venom from the parasitic species euplectrus comstockii howard (hymenoptera: eulophidae) with the pathogenicity of autographa californica (speyer) (lepidoptera: noctuidae) mnpv baculovirus (acmnpv) was tested in third and fourth instar larvae of trichoplusia ni (hübner) (lepidoptera: noctuidae). the presence of acmnpv did not alter the ability of the venom to arrest ecdysis in t. ni larvae. the presence of the venom delayed the rate of viruses by acmnpv but increased the ...	1999	10631800
high frequency recombination between homologous baculoviruses in cell culture.	the frequency of recombination between homologous baculoviruses was investigated in cell culture upon coinfection with two autographa californica nucleopolyhedrovirus (acmnpv) recombinants. these parental recombinants differed at two loci, separated by 20 kb, each carrying a different marker. the progeny recombinants generated by homologous recombination were easily distinguishable by the presence or absence of these markers. the mean percentage of the newly generated recombinants relative to a ...	2000	10664413
infectivity, speed of kill, and productivity of a baculovirus expressing the itch mite toxin txp-1 in second and fourth instar larvae of trichoplusia ni.	a cdna clone of the gene coding for the paralytic neurotoxin (tox34) from the female straw itch mite, pyemotes tritici, was created by rt-pcr and inserted into the genome of the autographa californica nucleopolyhedrovirus (acmnpv) under the control of the acmnpv p10 promoter. this recombinant virus, actox34.4, caused a rigid paralysis in infected larvae. the infectivity of actox34.4 was compared to the wild-type parent strain, acmnpv-c6, in second and fourth instar larvae of the cabbage looper, ...	2000	10753599
mutagenesis of the active site coding region of the autographa californica nucleopolyhedrovirus chia gene.	the chitinase of autographa californica nucleopolyhedrovirus (acmnpv) is required for the characteristic liquefaction of baculovirus-infected insect larvae. alignments of the putative active sites of a range of chitinases revealed two highly conserved residues, glutamate and aspartate, which have been proposed to constitute the catalytic residues of the active site. these residues were mutated in the acmnpv chitinase. three recombinant viruses were generated, acchia(d311g), acchia(e315g) and acc ...	2000	10769084
expression of the green fluorescent protein carried by autographa californica multiple nucleopolyhedrovirus in insect cell lines.	a recombinant acmnpv containing the green fluorescent protein (gfp) gene under the polyhedrin promoter (polh) was used to investigate the expression of the gfp gene as well as the production of recombinant extracellular virus in 14 continuous insect cell lines, including heliothis virescens (bcirl-hv-am1), helicoverpa zea (bcirl-hz-am1), anticarsia gemmatalis (bcirl-ag-am1), trichoplusia ni (tn-cl1), spodoptera frugiperda (iplb-sf21), spodoptera exigua (bcirl/amcy-se-e1 and bcirl/amcy-se-e5), bo ...	2000	10777062
diagnosis of penaeus monodon-type baculovirus by pcr and by elisa of occlusion bodies.	the black tiger prawn penaeus monodon is a valuable aquaculture product in taiwan. two specific diagnostic methods were established for p. monodon-type baculovirus, one using polymerase chain reaction (pcr) technology and the other enzyme-linked immunosorbent assay (elisa) technology. monodon-type baculovirus (mbv) was purified by sucrose gradient centrifugation from occlusion bodies of mbv-infected postlarvae of p. monodon. mbv dna was subsequently purified from the occlusion bodies and its pre ...	2000	10782342
differential activity of two non-hr origins during replication of the baculovirus autographa californica nuclear polyhedrosis virus genome.	the identification of potential baculovirus origins of replication (ori) has involved the generation and characterization of defective interfering particles that contain major genomic deletions yet retain their capability to replicate by testing the replication ability of transiently transfected plasmids carrying viral sequences in infected cells. so far, there has not been any evidence to demonstrate the actual utilization of these putative origins in autographa californica multinucleocapsid nu ...	2000	10799593
the autographa californica nuclear polyhedrosis virus p143 gene encodes a dna helicase.	the p143 protein of autographa californica nuclear polyhedrosis virus is essential for replication of viral dna. to determine the function of p143, the protein was purified to near homogeneity from recombinant baculovirus-infected cells that overexpress p143. atpase activity copurified with p143 protein during purification and also during gel filtration at a high salt concentration. the atpase activity did not require the presence of single-stranded dna, but was stimulated fourfold by the additi ...	2000	10799604
midgut-based resistance of heliothis virescens to baculovirus infection mediated by phytochemicals in cotton.	the decrease in susceptibility to polyhedrosis disease when heliothis virescens larvae feed on cotton is profound, limiting the utility of baculoviruses for controlling noctuids on this important crop. we observed that the mortalities of h. virescens larvae challenged with a reporter-gene construct of autographa californica m nucleopolyhedrovirus (acmnpv-hsp70/lacz) and fed either lettuce or artificial diet were approximately 2.5-fold higher than that of cotton-fed insects. this decrease in susc ...	2000	10802113
the expression and biologic effects of ovine interleukin-4 on t and b cell proliferation.	using the reverse-transcriptase polymerase chain reaction (rt-pcr), cdna encoding ovine (ov) interleukin-4 (ovil-4) was generated from mitogen-stimulated peripheral blood mononuclear cells (pbmc). two identical clones generated from separate rt-pcr reactions differed from a published ovil-4 sequence, although they had a high degree of identity with the bovine and human homologs. we show by sequence analysis that the ovil-4 cdna retained the four alpha-helix structure and disulfide bonds identifi ...	2000	10805377
the trichoplusia ni granulovirus helicase is unable to support replication of autographa californica multicapsid nucleopolyhedrovirus in cells and larvae of t. ni.	baculovirus dna helicases are essential for replication and are determinants of host range. helicases of autographa californica multicapsid nucleopolyhedrovirus (acmnpv) and trichoplusia ni granulovirus (tngv) differ markedly, although both viruses replicate efficiently in the cabbage looper, t. ni. to determine whether the tngv helicase (p137) could support replication of acmnpv in t. ni cells or larvae, the native acmnpv helicase gene (p143) was disrupted and substituted with p137. p137 did no ...	2000	10811943
protein requirements for assembly of virus-like particles of junonia coenia densovirus in insect cells.	the coding sequences of four overlapping polypeptides starting at four different in-frame aug codons and co-terminating at the stop codon of the cap gene of junonia coenia densovirus (jcdnv) were inserted under the control of the p10 promoter of autographa californica nucleopolyhedrovirus (acmnpv) to generate acmnpv-vp1 (four polypeptides), acmnpv-vp2 (three polypeptides), acmnpv-vp3 (two polypeptides), and acmnpv-vp4 (one polypeptide) recombinant viruses. in all cases, infection of spodoptera f ...	2000	10811945
identification of the lymantria dispar nucleopolyhedrovirus envelope fusion protein provides evidence for a phylogenetic division of the baculoviridae.	the complete genome sequences of a number of diverse members of the baculoviridae including both nucleopolyhedroviruses (npvs) and granuloviruses (gvs) revealed that they lack a homolog of gp64, the envelope fusion protein of the budded form of autographa californica multinucleocapsid npv (acmnpv) and its close relatives. computer-assisted analyses of the genome of one of these viruses, lymantria dispar mnpv (ldmnpv), revealed a single open reading frame (ld130) whose product had the predicted p ...	2000	10846096
potato leafroll virus protein p1 contains a serine proteinase domain.	the multi-domain potato leafroll virus replicase protein p1 was expressed in insect cells from the polyhedrin promoter of autographa californica nucleopolyhedrovirus. using antisera raised against p1, it was shown that p1 was cleaved near the vpg in insect cells in a manner similar to that in plant cells, to produce a approximately 27 kda c-terminal fragment. furthermore, it was shown that the proposed serine proteinase-like domain within p1 is responsible for this processing and that this can o ...	2000	10859393
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 ...	2000	10859396
expression of highly controllable genes in insect cells using a modified tetracycline-regulated gene expression system.	a modified tetracycline-responsive expression system (tres) for use in insect cells was developed. the tres contains two components: one encodes a tetracycline-controllable transactivator (tta) and the other contains a tet operator dna sequence to drive the luciferase gene. our results show that the human cytomegalovirus (cmv) promoter, an essential part for strong tta expression in mammalian system, was not functional in insect cells. thus further modifications were required. functional tta was ...	2000	10862988
characterization of a baculovirus alkaline nuclease.	all baculovirus genomes sequenced to date encode a homolog of an alkaline nuclease that has been characterized in the herpesviridae. in this report we describe the characterization of the alkaline nuclease (an) homolog of the autographa californica multinucleocapsid nucleopolyhedrovirus (acmnpv) (open reading frame 133). his-tagged an constructs were expressed in recombinant baculoviruses and affinity purified, and then their enzymatic activity was characterized. an was found to degrade linear d ...	2000	10864651
expanding baculovirus surface display. modification of the native coat protein gp64 of autographa californica npv.	to create a tool for eukaryotic surface display, this approach is aimed at demonstrating a direct modification of the native envelope protein gp64 of autographa californica npv without disturbing viral infectivity. short affinity-tag peptides, the biotin mimic streptagii, and the gp41 amino-acid motif eldkwa of hiv-1, specific for the human monoclonal antibody 2f5, were engineered into the baculovirus major coat protein gp64 and presented on the viral surface. two different streptag peptides wer ...	2000	10866803
site-directed mutagenesis of autographa californica nucleopolyhedrovirus (acnpv) polyhedrin: effect on polyhedron structure.	amino acids lys34, his36, and phe37 were substituted by pcr-mediated, site-directed mutagenesis for three trp's in the acnpv polyhedrin sequence. phase contrast microscopy revealed refringent, amorphous polyhedra in the nuclei of infected cells. electron microscopy confirmed a great variation in form and size of the mutated polyhedra. although crystallization of the mutated polyhedrin occurred, it was irregular within each polyhedron. virion occlusion was also severely affected. virions were par ...	2000	10893160
enhancement in activity of homologous and heterologous baculoviruses infectious to beet armyworm (lepidoptera: noctuidae) by an optical brightener.	the nuclear polyhedrosis virus (npv) from the beet armyworm, spodoptera exigua (hübner) (semnpv), was the most active virus tested against the beet armyworm (lc50 = 4.1 pibs/mm2), followed by nuclear polyhedrosis viruses from the alfalfa looper, autographa californica (speyer) (acmnpv; lc50 = 92.6 pibs/mm2), and the celery looper, anagrapha falcifera (kirby) (afmnpv; lc50 = 195.7 pibs/mm2). in the case of the nuclear polyhedrosis virus from the bollworm, helicoverpa armigera (hübner), lc50s coul ...	2000	10902301
influence of baculovirus-host cell interactions on complex n-linked glycosylation of a recombinant human protein.	the conditions required for mammalian-type complex n-linked glycosylation of human proteins produced in insect cells with the baculovirus expression vector system were investigated. marked alterations to n-linked glycosylation of human placental secreted alkaline phosphatase (seap) were observed with different baculovirus species, insect cell lines, and cell culture media. when a recombinant autographa californica nucleopolyhedrovirus (acmnpv) was used to produce seap in trichoplusia ni (tn-4h) ...	2000	10933841
the autographa californica nucleopolyhedrovirus ie-1 protein complex has two modes of specific dna binding.	missing contact footprinting with formic acid as a modifying reagent was used to examine specific ie-1 binding contacts to double-stranded oligonucleotides that contained either a consensus hr repeat sequence or a sequence from the pe38 promoter, which is down regulated by ie-1. the hr repeat sequences contain two consensus ie-1 binding motifs (ibms) flanking a central ecori site that are oriented in opposite directions with respect to each other. ie-1 was found to contact regions including both ...	2000	10936100
baculoviral display of functional scfv and synthetic igg-binding domains.	viral vectors displaying specific ligand binding moities such as scfv fragments or intact antibodies hold promise for the development of targeted gene therapy vectors. in this report we describe baculoviral vectors displaying either functional scfv fragments or the synthetic z/zz igg binding domain derived from protein a. display on the baculovirus surface was achieved via fusion of the scfv fragment or z/zz domain to the n-terminus of gp64, the major envelope protein of the autographa californi ...	2000	10944446
further characterization of the gonad-specific virus of corn earworm, helicoverpa zea.	the gonad-specific virus (gsv) is a dna virus infecting the reproductive tracts of adults of both sexes of the corn earworm, helicoverpa zea, causing severe tissue deformities leading to sterility. atypical occlusion bodies containing large concentrations of virions embedded in a granular matrix were seen in the lumen of the oviduct and the bursa copulatrix of infected females. the virus, transmitted by both sexes, was successfully propagated in vivo and in tissue culture. the gsv genome is abou ...	2000	10963397
abnormal formation of polyhedra resulting from a single mutation in the polyhedrin gene of autographa californica multicapsid nucleopolyhedrovirus.	a spontaneous mutant that produces a single abnormally large cubic polyhedron per infected cell was isolated from a polyhedra-positive recombinant autographa californica multicapsid nucleopolyhedrovirus (acmnpv). both wild-type and mutant virus produce two forms of virus particles, budded virions and occluded virions. however, occluded virions are not found within the polyhedra of cells infected with mutant virus, as with the wild-type virus. these large cubic polyhedra do not have the typical l ...	2000	10963398
impact of recombinant baculovirus field applications on a nontarget heliothine parasitoid, microplitis croceipes (hymenoptera: braconidae).	the kill times of two viruses infectious to the heliothine pest complex indigenous to texas cotton have been significantly reduced by expressing a scorpion toxin gene. autographa californica nucleopolyhedrovirus (npv) and helicoverpa zea npv express the toxin only in permissive lepidopteran hosts. the toxin, however, could indirectly harm members of upper trophic levels that feed upon and parasitize infected larvae producing the toxin. in this study, the effects of recombinant and wild-type viru ...	2000	10985020
virus morphogenesis of helicoverpa armigera nucleopolyhedrovirus in helicoverpa zea serum-free suspension culture.	helicoverpa armigera single nucleopolyhedrovirus (hasnpv) replication in helicoverpa zea serum-free suspension culture was studied in detail and the sequence of virus morphogenesis was determined by transmission electron microscopy. by 16 h post-infection (p.i.), virus replication was observed in the virogenic stroma by the appearance of nucleocapsids. polyhedron formation was detected by 24 h p.i. and the polyhedron envelope (pe) was completely formed by 72 h p.i. pe morphogenesis of hasnpv is ...	2000	10993944
effects of long- and short-term passage of insect cells in different culture media on baculovirus replication.	two insect cell lines that had been maintained in both serum-free (sfm) and serum-containing (scm) media for over 5 years were each tested for their ability to replicate baculovirus. the gypsy moth cell line, iplb-ldeita (ld), produced similar (not statistically different) amounts of gypsy moth nucleopolyhedrovirus (ldmnpv) occlusion bodies (obs) in the two media (serum-free ex-cell 400 and tc-100 with 9% (v/v) fetal bovine serum, scm(1)) but produced more of the autographa californica nucleopol ...	2000	11023743
effects of incubation temperature on the dose-survival time relationship of trichoplusia ni larvae infected with autographa californica nucleopolyhedrovirus.	the interaction between virus multiplication and host development was studied by determining the survival time of trichoplusia ni larvae inoculated with a wide range of doses of autographa californica nucleopolyhedrovirus and incubated at five different temperatures spanning the biologically relevant range. the results support earlier findings that the course of baculovirus infection follows the mathematical description of the birth-death model. both in vivo rate of virus increase and larval gro ...	2000	11023746
presentation of antigenic sites from foot-and-mouth disease virus on the surface of baculovirus and in the membrane of infected cells.	we describe the construction of recombinant baculoviruses displaying on their surface and in the membrane of infected cells the small, immunodominant antigenic site (site a) or the large polyprotein (p1) coding for the four structural proteins of foot-and-mouth disease virus (fmdv). the coding sequences were inserted in the amino-terminus of gp64, the major glycoprotein of the baculovirus autographa californica nuclear polyhedrosis virus (acnpv). following infection of insect cells with the reco ...	2000	11043943
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 ...	2001	11125177
comparative analysis of the granulin regions of the phthorimaea operculella and spodoptera littoralis granuloviruses.	the nucleotide sequence of two cloned restriction fragments encompassing the granulin genes from the granuloviruses of the potato tuber moth, phthorimaea operculella, phopgv, and the egyptian cotton leaf worm, spodoptera littoralis, spligv, have been determined. although both viruses are able to infect the same ph. operculella cell line, their granulins do not cluster in the same phylogenetic branches. phopgv ganulin is closely related to cydia pomonella gv (cpgv) and cryptophlebia leucotreta gv ...	2000	11129630
identification of insertion and deletion genes in autographa californica nucleopolyhedrovirus variants isolated from galleria mellonella, spodoptera exigua, spodoptera litura and xestia c-nigrum.	the genomic dna of four autographa californica multinucleocapsid nucleopolyhedrovirus (acmnpv) variants isolated from galleria mellonella, spodoptera exigua, spodoptera litura and xestia c-nigrum was analyzed in comparison with the acmnpv e2 strain. restriction endonuclease analysis revealed a deletion and an insertion in collinear regions of the four variants. polymerase chain reaction analysis indicated that, in the four variants, the deletion occurred in the region corresponding to acmnpv c6 ...	2000	11129632
central role of hemocytes in autographa californica m nucleopolyhedrovirus pathogenesis in heliothis virescens and helicoverpa zea.	autographa californica m nucleopolyhedrovirus (acmnpv) can infect and kill a wide range of larval lepidopteran hosts, but the dosage required to achieve mortal infection varies greatly. using a reporter gene construct, we identified key differences between acmnpv pathogenesis in heliothis virescens and helicoverpa zea, a fully permissive and a semipermissive host, respectively. even though there was more than a 1,000-fold difference in the susceptibilities of these two species to mortal infectio ...	2001	11134313
production of biologically active recombinant bovine interferon-gamma by two different baculovirus gene expression systems using insect cells and silkworm larvae.	the full-length bovine interferon-gamma (bifn-gamma) cdna, including the secretion signal peptide coding region was recloned into baculovirus transfer vectors pacym1 and pbm050. these vectors were co-transfected with autographa californica nuclear polyhedrosis virus (acnpv) or bombyx mori nuclear polyhedrosis virus (bmnpv) dna into spodoptera frugiperda cells (sf21ae) and bombyx mori cells (bmn), respectively. the recombinant viruses, named acbifn-gamma and bmbifn-gamma, were then recovered. rec ...	2001	11145838
p34.8 (gp37) is not essential for baculovirus replication.	previous reports have indicated that p34.8 (gp37) may be essential for the replication of autographa californica nucleopolyhedrovirus (acmnpv) because no virus with inactivated p34.8 was isolated. we have ascertained the requirement for this gene by attempting to inactivate it with a large insertion [the gene encoding gfp (green fluorescent protein)] or by deleting all the conserved domains from the open reading frame (orf). the gene encoding gfp was inserted into the not:i site of the p34.8 orf ...	2001	11161266
high-level expression of human acidic fibroblast growth factor and basic fibroblast growth factor in silkworm (bombyx mori l.) using recombinant baculovirus.	a hybrid of autographa californica nuclear polyhedrosis virus and bombyx mori nuclear polyhedrosis virus, which is infectious to both spodoptera frugiperda and bombyx mori, was prepared in our previous study. two recombinant hybrid baculoviruses, carrying cdnas of human acidic and basic fibroblast growth factors, respectively, were successfully constructed in this study, for the large-scale production of human afgf and bfgf using silkworm as host. these recombinant viruses were used to inoculate ...	2001	11162406
diagnostic immunoassays for tick-borne encephalitis virus based on recombinant baculovirus protein expression.	the baculovirus expression system that utilizes autographa californica nuclear polyhedrosis virus was used to express the highly antigenic envelope protein e of a tick-borne encephalitis (tbe) complex virus, as well as a c-terminally truncated form of protein e (etr). the recombinant proteins were produced with a histidine-tag at their carboxy-terminus. protein purification by nickel agarose chromatography resulted in high concentrations of pure etr protein, but only poor yields of e protein. th ...	2001	11164488
[expression of 2.1 kb enhancin gene fragment from helicoverpa armigera granulosis virus in escherichia coli].	the 2.1 kb fragment of enhancin gene from helicoverpa armigera granulosis virus was inserted into vector pqe-30 and expressed successfully in e. coli m15(prep4). the synergy of expression product(p78) on acmnpv against the larvae of plutella xylostella was also studied. the results indicated that the percentage of correct mortality of the larvae increased 27.88%-32.92% in 10 post-infection days.	2000	11191765
a gp64-null baculovirus pseudotyped with vesicular stomatitis virus g protein.	the autographa californica multiple nucleopolyhedrovirus (acmnpv) gp64 protein is an essential virion protein that is involved in both receptor binding and membrane fusion during viral entry. genetic studies have shown that gp64-null viruses are unable to move from cell to cell and this results from a defect in the assembly and production of budded virions (bv). to further examine requirements for virion budding, we asked whether a gp64-null baculovirus, vac(64-), could be pseudotyped by introdu ...	2001	11222677
behavior of a recombinant baculovirus in lepidopteran hosts with different susceptibilities.	insect pathogens, such as baculoviruses, that are used as microbial insecticides have been genetically modified to increase their speed of action. nontarget species will often be exposed to these pathogens, and it is important to know the consequences of infection in hosts across the whole spectrum of susceptibility. two key parameters, speed of kill and pathogen yield, are compared here for two baculoviruses, a wild-type autographa californica nucleopolyhedrovirus (acnpv), acnpv clone c6, and a ...	2001	11229903
dynamic nuclear localization of the baculovirus proteins ie2 and pe38 during the infection cycle: the promyelocytic leukemia protein colocalizes with ie2.	the early gene products ie2 and pe38 of autographa californica multicapsid nuclear polyhedrosis virus localize to distinct nuclear domains after transient expression. here, the nuclear localization pattern and the putative association with cellular proteins have been determined during virus infection to shed light on the functional significance of the nuclear domains. ie2 was always localized to distinct nuclear structures while pe38 was partly present in nuclear dots. confocal imaging indicated ...	2001	11262179
actin rearrangement-inducing factor of baculoviruses is tyrosine phosphorylated and colocalizes to f-actin at the plasma membrane.	in previous studies we have identified actin rearrangement-inducing factor 1 as an early gene product of autographa californica multicapsid nuclear polyhedrosis virus that is involved in the remodeling of the actin cytoskeleton. we have constructed viral recombinants with a mutated arif-1 open reading frame that confirm the causal link of arif-1 expression and the actin rearrangement observed as accumulation of f-actin at the plasma membrane at 3 to 7 h postinfection. infection with arif mutant ...	2001	11264366
novel sp family-like transcription factors are present in adult insect cells and are involved in transcription from the polyhedrin gene initiator promoter.	we earlier documented the involvement of a cellular factor, polyhedrin (polh) promoter-binding protein, in transcription from the autographa californica nuclear polyhedrosis virus polh gene promoter. sequences upstream of the polh promoter were found to influence polh promoter-driven transcription. analysis of one such region, which could partially compensate for the mutated polh promoter and also activate transcription from the wild-type promoter, revealed a sequence (acsp) containing a caccc m ...	2001	11294840
autographa californica baculoviruses with large genomic deletions are rapidly generated in infected insect cells.	defective interfering baculoviruses (dis) lack considerable portions of the genome, interfere with the replication of helper virus, and cause the so-called "passage-effect" during serial passaging in insect cells and in bioreactor configurations. we investigated their origin by (nested) pcr and demonstrated that dis lacking approximately 43% (d43) of their dna are present in low-passage autographa californica multicapsid nucleopolyhedrovirus (acmnpv)-e2 virus stocks and in polyhedra, but not in ...	2001	11312669
combination treatment for osteosarcoma with baculoviral vector mediated gene therapy (p53) and chemotherapy (adriamycin).	the insect baculovirus autographa californica multiple nuclear polyhedrosis virus (acmnpv) has been evaluated as a vector for gene delivery to human tumor cells. a human osteogenic sarcoma cell line, saos-2, was found to be highly susceptible to infection with a baculoviral vector, with nearly 100% of saos-2 cells being able to express a lacz reporter gene after a brief exposure to the virus at a m.o.i. of 30 pfu/cell. the production of beta-galactosidase protein was 18-times greater than that i ...	2001	11322486
cloning and characterization of an inhibitor of apoptosis protein (iap) from bombyx mori.	we cloned a novel inhibitor of apoptosis protein (iap) family member, bmiap, from bombyx mori bmn cells. bmiap contains two baculoviral iap repeat (bir) domains followed by a ring domain. bmiap shares striking amino acid sequence similarity with lepidopteran iaps, sfiap and tniap, and with two baculoviral iaps, cpiap and opiap, suggesting evolutionary conservation. bmiap blocks programmed cell death (apoptosis) in spodoptera frugiperda sf-21 cells induced by p35 deficient autographa californica ...	2001	11341966
developments in the use of baculoviruses for the surface display of complex eukaryotic proteins.	the ability to couple genotype to phenotype has proven to be of immense value in systems such as phage display and has allowed genes encoding novel functions to be selected directly from complex libraries. however, the complexity of many eukaryotic proteins places a severe constraint on successful display in escherichia coli. this restriction could be resolved if a eukaryotic virus could be similarly engineered for display purposes. preliminary data have suggested that the baculovirus autographa ...	2001	11356285
specific binding of baculoviruses displaying gp64 fusion proteins to mammalian cells.	viral vectors displaying specific ligand binding moieties have raised an increasing interest in the area of targeted gene therapy. in this report, we describe baculovirus vectors displaying either a functional single chain antibody fragment (scfv) specific for the carcinoembryonic antigen (cea) or the synthetic igg binding domains (zz) derived from protein a of staphylococcus aureus. in addition, the vectors were engineered to incorporate a reporter gene encoding the enhanced green fluorescent p ...	2001	11396970
a new continuous cell line from larval hemocytes of spodoptera litura (f.).	a new cell line has been established from larval hemocytes of the moth, s. litura (tobacco cut worm). it took 147 days to form a monolayer and one year for the first 17 passages. at present, the culture is at 86th passage level and is designated niv-su-1095. three cell types could be distinguished, viz. plasmatocytes (53%), prohemocytes (36%) and granular hemocytes (11%). the chromosome number was very high, 74% metaphase cells showed more than 100 chromosomes. the cells could be cryopreserved. ...	2000	11411040
expression and one-step purification of intracellular human prolactin in insect cells.	human prolactin was expressed in insect culture cells by recombinant baculoviruses carrying prolactin gene cdna placed under the transcriptional control of polyhedrin gene promoter of autographa californica nuclear polyhedrosis virus. preliminary results of recombinant human prolactin expression as extracellular as well as intracellular product of baculovirus expression system were presented at the febs meeting in nice, france, in 1999 (abstracts, p. 288). in the present work prolactin was expre ...	2001	11437600
a study of the autographa californica multiple nucleopolyhedrovirus odv envelope protein p74 using a gfp tag.	the autographa californica multiple nucleopolyhedrovirus (acmnpv) protein p74 is associated with the occlusion-derived virus (odv) envelope. p74 is essential for oral infectivity of odv and has been proposed to play a role in midgut attachment and/or fusion. in this study, p74 protein was expressed in-frame with green fluorescent protein (gfp) to create a p74-gfp chimera. the c-terminal gfp portion of the chimera facilitated visualization of the trafficking of p74 in baculovirus-infected spodopt ...	2001	11514740
expression and localization of lef-11 in autographa californica nucleopolyhedrovirus-infected sf9 cells.	the autographa californica multicapsid nucleopolyhedrovirus (acmnpv) lef-11 gene was found previously to be necessary to support optimal levels of transient expression from an acmnpv late promoter. the lef-11 gene is unusual in that it overlaps both upstream (orf38) and downstream (pp31) genes. in this study, the expression and cellular localization of lef-11 were examined. the lef-11 transcripts were detected from 4 to 36 h post-infection (p.i.). the 1.5 kb lef-11 mrna initiates 196 nt upstream ...	2001	11514741
production of selected baculoviruses in newly established lepidopteran cell lines.	one key to the in vitro mass production of baculoviruses is the development of insect cell lines capable of producing high levels of extracellular virus (ecv) and/or occlusion bodies (obs). for this study, 34 newly established cell lines from 10 lepidopteran species were screened for their ability to produce ecv and obs from a variety of baculoviruses. the selected baculoviruses included: the alfalfa looper virus (acmnpv); the celery looper virus (afmnpv); the velvetbean caterpillar virus (agmnp ...	2001	11515971
sequence analysis of the spodoptera litura multicapsid nucleopolyhedrovirus genome.	the complete spodoptera litura multicapsid nucleopolyhedrovirus (spltmnpv) genome contained 139,342 bp with a g+c content of 42.7%, and 141 putative open reading frames (orfs) or genes of 150 nucleotides or greater that showed minimal overlap. ninety-six orfs had homologues in autographa californica multicapsid nucleopolyhedrovirus (acmnpv), 16 had homologues in other baculoviruses, and 29 were unique to spltmnpv. the homologues of ubiquitin and gp37 are fused in spltmnpv. the genome lacked a ho ...	2001	11531416
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 ...	2001	11556402
the complete sequence of the cydia pomonella granulovirus genome.	the nucleotide sequence of the dna genome of cydia pomonella granulovirus (cpgv) was determined and analysed. the genome is composed of 123500 bp and has a g+c content of 45.2%. it contains 143 orfs of 150 nucleotides or more that show minimal overlap. one-hundred-and-eighteen (82.5%) of these putative genes are homologous to genes previously identified in other baculoviruses. among them, 73 are homologous to genes of autographa californica nucleopolyhedrovirus (acmnpv), whereas 108 and 98 are h ...	2001	11562546
investigation of sequential behavior of carboxyl protease and cysteine protease activities in virus-infected sf-9 insect cell culture by inhibition assay.	proteases produced during the culture of spodoptera frugiperda sf-9 cells infected with autographa californica nuclear polyhedrosis virus (acnpv) were assayed with various protease inhibitors. this inhibitory analysis revealed that: (1) carboxyl and cysteine proteases were predominantly produced by the insect cells infected with recombinant acnpv, the gene of which encoded a variant of green fluorescent protein in a portion of the polyhedrin gene of the baculovirus, and (2) the protease activity ...	2001	11601624
effects of deletion and overexpression of the autographa californica nuclear polyhedrosis virus fp25k gene on synthesis of two occlusion-derived virus envelope proteins and their transport into virus-induced intranuclear membranes.	partial deletions within autographa californica open reading frame 61 (fp25k) alter the expression and accumulation profile of several viral proteins and the transport of occlusion-derived virus (odv)-e66 to intranuclear membranes during infection (s. c. braunagel et al., j. virol. 73:8559-8570, 1999). here we show the effects of a full deletion and overexpression of fp25k on the transport and expression of two odv envelope proteins, odv-e66 (e66) and odv-e25 (e25). deletion and overexpression o ...	2001	11602724
persistence of an occlusion-negative recombinant nucleopolyhedrovirus in trichoplusia ni indicates high multiplicity of cellular infection.	we use data from the serial passage of co-occluded recombinant autographa californica nuclear polyhedrosis virus (acmnpv) to estimate the viral multiplicity of infection of cells within infected insects. co-occlusion, the incorporation of wild-type and mutant virus genomes in the same occlusion body, has been proposed as a strategy to deliver genetically modified viruses as insecticides in a way that contains their spread in the environment. it may also serve as a means whereby naturally occurri ...	2001	11679346
deletions in the ac-iap1 gene of the baculovirus acmnpv occur spontaneously during serial passage and confer a cell line-specific replication advantage.	the psti-i region of the autographa californica multicapsid nucleopolyhedrovirus (acmnpv) genome was previously shown to be a frequent target of spontaneous deletions during serial virus passage in tn-368 cells (kumar and miller, virus res. 7 (1987) 335). analysis of two of these serial passage mutants showed that a portion of the ac-iap1 gene was deleted. to directly test the effect of loss of ac-iap1, three different deletions in ac-iap1 were introduced into recombinant viruses and the ability ...	2001	11682127
identification of bv/odv-c42, an autographa californica nucleopolyhedrovirus orf101-encoded structural protein detected in infected-cell complexes with odv-ec27 and p78/83.	orf101 is a late gene of autographa californica nucleopolyhedrovirus (acmnpv). it encodes a protein of 42 kda which is a component of the nucleocapsid of budded virus (bv) and occlusion-derived virus (odv). to reflect this viral localization, the product of orf101 was named bv/odv-c42 (c42). c42 is predominantly detected within the infected-cell nucleus: at 24 h postinfection (p.i.), it is coincident with the virogenic stroma, but by 72 h p.i., the stroma is minimally labeled while c42 is more u ...	2001	11711623
novel baculovirus dna elements strongly stimulate activities of exogenous and endogenous promoters.	a dna sequence upstream from the polyhedrin gene of baculovirus autographa californica nucleopolyhedrovirus (acmnpv) was found to activate strongly the expression of full or minimal promoters derived from acmnpv and other sources. promoters tested included the minimal cmv (cmvm) promoter from human cytomegalovirus, the full heat shock 70 promoter from drosophila, and the minimal p35 promoter from baculovirus. deletion and mutagenesis analyses showed that this functional polyhedrin upstream (pu) ...	2002	11741907
identification of apoptosis-inhibiting gene in leucania separata nuclear polyhedrosis virus.	a novel gene lsp40 from leucania separata nuclear polyhedrosis virus (lsnpv) which was homologous to the p35 gene from autographa californica nuclear polyhedrosis virus (acnpv) was localized in the ecorv-5.5 kb fragment of lsnpv genome dna and was sequenced. the open reading frame (orf) of lsp40 was 906 bp long and encoded an approximately 40 kda peptide consisting of 302 amino acid residues. the isp40 shares 80.4% and 70.4% identity of nucleotide and amino acid sequence, respectively, to the ac ...	2001	11765917
assessment of foreign protein production by recombinant heliothis (helicoverpa) armigera entomopoxviruses in spodoptera frugiperda cells.	this report describes the first production of recombinant forms of heliothis (helicoverpa) armigera entomopoxvirus (haepv). these haepvs are engineered at either the spheroidin or fusolin locus, to produce the green fluorescent marker protein (gfp). the growth properties of these recombinant haepvs, in comparison to the parental haepv, were assessed in cultured spodoptera frugiperda sf9 cells. additionally, gfp production by these recombinant haepvs was compared to that of a gfp-expressing recom ...	2002	11807239
biosynthesis and localization of the autographa californica nuclear polyhedrosis virus 25k gene product.	mutations of the acmnpv 25k gene are associated with the "few polyhedra" phenotype (m. j. fraser et al., 1983, j. virol. 47, 287-300; b. beames and m. d. summers, 1989, virology 168, 344-353). polyclonal antisera was produced and used to investigate the time course of expression and localization of the 25k protein in infected cells. western blot analysis detected 25k protein in both cytosolic and nuclear extracts from 18-24 hr p.i. through 96 hr p.i. and also in purified viral occlusions, but no ...	1995	11831709
passage of autographa californica nuclear polyhedrosis virus through the midgut epithelium of spodoptera exigua larvae.	a special recombinant of autographa californica multicapsid nuclear polyhedrosis virus (acnpv) was designed to study the early histopathological events of baculovirus infection in spodoptera exigua larvae. this recombinant contained a drosophila melanogaster heat shock 70 promoter driving an escherichia coli beta-galactosidase (lac-z) reporter gene to monitor the presence of early viral gene expression and a second reporter gene, the e. coli beta-glucuronidase (gus) gene, under control of the ve ...	1995	11831715
rat monoclonal antibodies differentiating between the epstein-barr virus nuclear antigens 2a (ebna2a) and 2b (ebna2b).	rat monoclonal antibodies were produced against the c-terminus of epstein-barr virus nuclear antigens 2a (ebna2a) and 2b (ebna2b) expressed as bacterial trpe fusion proteins. the initial screening was performed using a soluble bacterial extract containing the fusion proteins. positive hybridomas were confirmed by immunofluorescence on sf158 (spodoptera frugiperda) insect cells infected with recombinant baculovirus (autographa californica nuclear polyhedrosis virus) and expressing the complete eb ...	1995	11831716
intron-dependent stimulation of marker gene expression in cultured insect cells.	we tested in a systematic fashion the effect of an intron on the level of luciferase expression in cultured c6/36 aedes albopictus cells. the intron was inserted in both orientations, upstream and downstream of the luciferase coding region in two different luciferase expression vectors. the two parental luciferase expression vectors differed only in their promoters, one containing the drosophila melanogaster actin5c promoter and the other the autographa californica nuclear polyhedrosis virus hr5 ...	2002	11841506
partial redistribution of the autographa californica nucleopolyhedrovirus chitinase in virus-infected cells accompanies mutation of the carboxy-terminal kdel er-retention motif.	during virus infection of insect cells, the autographa californica nucleopolyhedrovirus chitinase is localized primarily within the endoplasmic reticulum (er), which is consistent with the presence of a carboxy-terminal er retention motif (kdel). release of chitinase into the extracellular medium appears to be concomitant with terminal cell lysis, rather than by active secretion. in this study, we have shown that mutation of the kdel motif induces a partial redistribution of the chitinase at bot ...	2002	11842263
characterization of a truncated soluble form of the baculovirus (acmnpv) major envelope protein gp64.	a truncated tagged form of the autographica californica multiple nuclear polyhedrosis virus major surface glycoprotein, gp64, has been expressed using the baculovirus expression system and purified to homogeneity by immune-affinity chromatography. the protein, which is responsible for virus-cell fusion, was a trimer in solution and retained this oligomeric form at ph 5, the ph of fusion. circular dichroism spectroscopy indicated a protein with mixed alpha-helix and beta-sheet content that did no ...	2002	11858713
analysis of an autographa californica nucleopolyhedrovirus lef-11 knockout: lef-11 is essential for viral dna replication.	the autographa californica nucleopolyhedrovirus (acmnpv) lef-11 gene was previously identified by transient late expression assays as a gene important for viral late gene expression. the lef-11 gene was not previously identified as necessary for dna replication in transient origin-dependent plasmid dna replication assays. to examine the role of lef-11 in the context of the infection cycle, we generated a deletion of the lef-11 gene by recombination in an acmnpv genome propagated as a bacmid in e ...	2002	11861844
phenotypic and genotypic analysis of helicoverpa armigera nucleopolyhedrovirus serially passaged in cell culture.	rapid accumulation of few polyhedra (fp) mutants was detected during serial passaging of helicoverpa armigera nucleopolyhedrovirus (hasnpv) in cell culture. 100% fp infected cells were observed by passage 6. the specific yield decreased from 178 polyhedra per cell at passage 2 to two polyhedra per cell at passage 6. the polyhedra at passage 6 were not biologically active, with a 28-fold reduction in potency compared to passage 3. electron microscopy studies revealed that very few polyhedra were ...	2002	11907345
nuclear ie2 structures are related to viral dna replication sites during baculovirus infection.	the ie2 gene of autographa californica multicapsid nuclear polyhedrosis virus is 1 of the 10 baculovirus genes that have been identified as factors involved in viral dna replication. ie2 is detectable in the nucleus as one of the major early-expressed proteins and exhibits a dynamic localization pattern during the infection cycle (d. murges, i. quadt, j. schröer, and d. knebel-mörsdorf, exp. cell res. 264:219-232, 2001). here, we investigated whether ie2 localized to regions of viral dna replica ...	2002	11967334
analysis of an autographa californica multicapsid nucleopolyhedrovirus lef-6-null virus: lef-6 is not essential for viral replication but appears to accelerate late gene transcription.	the autographa californica multicapsid nucleopolyhedrovirus (acmnpv) lef-6 gene was previously shown to be necessary for optimal transcription from an acmnpv late promoter in transient late expression assays. in the present study, we examined the expression and cellular localization of lef-6 during the acmnpv infection cycle and generated a lef-6-null virus for studies of the role of lef-6 in the infection cycle. transcription of lef-6 was detected from 4 to 48 h postinfection, and the lef-6 pro ...	2002	11991978
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 ...	2002	11991988
pseudotyping autographa californica multicapsid nucleopolyhedrovirus (acmnpv): f proteins from group ii npvs are functionally analogous to acmnpv gp64.	gp64, the major envelope glycoprotein of budded virions of the baculovirus autographa californica multicapsid nucleopolyhedrovirus (acmnpv), is involved in viral attachment, mediates membrane fusion during virus entry, and is required for efficient virion budding. thus, gp64 is essential for viral propagation in cell culture and in animals. recent genome sequences from a number of baculoviruses show that only a subset of closely related baculoviruses have gp64 genes, while other baculoviruses ha ...	2002	11992001