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the autographa californica multiple nucleopolyhedrovirus ie0-ie1 gene complex is essential for wild-type virus replication, but either ie0 or ie1 can support virus growth.the immediate-early ie0-ie1 gene complex expresses the only baculovirus spliced gene that produces an alternate protein product. autographa californica multiple nucleopolyhedrovirus (acmnpv) ie1 is a potent transcriptional transactivator that is essential for viral replication in transient assays. ie1 contains 582 amino acids that are arranged into different domains, including an acidic activation domain at the n terminus, a dna binding domain, and an oligomerization domain at the c terminus. ie ...200515795248
spodoptera frugiperda resistance to oral infection by autographa californica multiple nucleopolyhedrovirus linked to aberrant occlusion-derived virus binding in the midgut.spodoptera frugiperda larvae are highly resistant to oral infection by autographa californica multiple nucleopolyhedrovirus (acmnpv) (ld(50), approximately 9200 occlusions), but extremely susceptible to budded virus within the haemocoel (ld(50), <1 p.f.u.). the inability of acmnpv occlusion-derived virus (odv) to establish primary infections readily within midgut cells accounts for a major proportion of oral resistance. to determine whether inappropriate binding of acmnpv odv to s. frugiperda mi ...200515831946
stimulation of cell motility by a viral fibroblast growth factor homolog: proposal for a role in viral pathogenesis.the autographa californica m nucleopolyhedrovirus (acmnpv) encodes a gene (open reading frame 32) with homology to vertebrate and invertebrate fibroblast growth factors (fgfs), key regulators of developmental processes affecting the growth, differentiation, and migration of many cell types. we studied the temporal regulation of the acmnpv fgf, vfgf, by northern (rna) blot hybridization; vfgf was transcribed as a 0.6-kb mrna at early times but as part of a 1.4-kb bicistronic mrna at late times. t ...200515892971
insecticidal effects of buthus occitanus tunetanus botit6 toxin expressed in escherichia coli and baculovirus/insect cells.botit6 is a neurotoxin polypeptide derived from the venom of the scorpion buthus occitanus tunetanus (bot). its mature form is composed of 62 amino acids. botit6 has been reported to be the most potent toxin from bot venom that has a strict selectivity for insects. such toxin may have potential as a potent animal-harmless tool against insects. using rt-pcr, we isolated and sequenced a cdna encoding 62 amino acid residues corresponding to the known amino acid sequence of botit6. we have expressed ...200516216259
functional characterization of bombyx mori nucleopolyhedrovirus late gene transcription and genome replication factors in the non-permissive insect cell line sf-21.we compared the abilities of late gene transcription and dna replication machineries of the baculoviruses autographa californica nucleopolyhedrovirus (acmnpv) and bombyx mori npv (bmnpv) in sf-21 cells, an insect-derived cell line permissive for acmnpv infection. it has been well established that 19 acmnpv late expression factors (lefs) stimulate substantial levels of late gene promoter activity in sf-21 cells. thus, we constructed a set of clones containing the bmnpv homologs of the acmnpv lefs ...200616442141
in vivo analysis of fibroin heavy chain signal peptide of silkworm bombyx mori using recombinant baculovirus as vector.in order to investigate the functional signal peptide of silkworm fibroin heavy chain (fibh) and the effect of n- and c-terminal parts of fibh on the secretion of fibh in vivo, n- and c-terminal segments of fibh gene were fused with enhanced green fluorescent protein (egfp) gene. the fused gene was then introduced into silkworm larvae and expressed in silk gland using recombinant acmnpv (autographa californica multiple nuclear polyhedrosis virus) as vector. the fluorescence of egfp was observed ...200616466694
antibody-based surface plasmon resonance detection of intact viral pathogen.surface plasmon resonance (spr) technique was used to directly detect an intact form of insect pathogen: the baculovirus, autographa californica multiple nuclear polyhedrosis virus (acmnpv). an spr sensor chip with three bio-functional layers was used to detect the intact acmnpv: amine-reactive crosslinker with a disulfide bond that chemisorbs to gold film, protein a, and a mouse igg monoclonal antibody raised against a surface protein of the target viral pathogen. a two-channel (reference & tes ...200616470580
expression of autographa californica multiple nucleopolyhedrovirus genes in mammalian cells and upregulation of the host beta-actin gene.the gene expression of autographa californica multiple nucleopolyhedrovirus (acmnpv) was examined in two types of mammalian cells, human hela14 and hamster bhk cells. dna microarray analysis followed by reverse transcription-pcr identified at least 12 viral genes transcribed in both hela14 cells and bhk cells inoculated with acmnpv. 5' rapid amplification of cdna ends was carried out to examine the transcriptional fidelity of these genes in hela14 cells. the transcription of ie-1, ie-0 and gp64 ...200616474145
analysis of a baculovirus lacking a functional viral fibroblast growth factor homolog.baculoviruses encode fibroblast growth factor (vfgf) homologs whose function during virus infection is unknown. we constructed a recombinant bacmid of autographa californica m nucleopolyhedrovirus (acmnpv) lacking a functional vfgf and characterized it in two insect cell lines. the kinetics of budded virus production were similar in the parental and vfgf-deficient viruses in both cell lines at both high and low multiplicities of infection. in addition, no obvious differences were observed betwee ...200616476460
dual mutations in the autographa californica nucleopolyhedrovirus fp-25 and p35 genes result in plasma-membrane blebbing in trichoplusia ni cells.spodoptera frugiperda cells infected with autographa californica nucleopolyhedrovirus (acmnpv) lacking a functional anti-apoptotic p35 protein undergo apoptosis. however, such mutants replicate normally in trichoplusia ni (tn-368) cells. an acmnpv plaque isolate (acdefrt) was identified during propagation of a virus deficient in p35 in tn-368 cells. this virus exhibited enhanced budded-particle formation in tn-368 cells, but was partially defective for polyhedra production in the same cells. vir ...200616476974
baculovirus etl promoter acts as a shuttle promoter between insect cells and mammalian cells.to identify a shuttle promoter that can mediate gene expression in both insect cells and mammalian cells to facilitate the development of a baculovirus vector-based mammalian cell gene delivery vehicle.200616490168
identification of spodoptera litura multicapsid nucleopolyhedrovirus orf97, a novel protein associated with envelope of occlusion-derived virus.open reading frame (orf) 97 of spodoptera litura multicapsid nucleopolyhedrovirus (splt97) is a member of 11k gene family (interpro database accession number: ipr009313) in insect viruses, potentially encoding 112 amino acids with a predicted molecular weight of 13.2 kda. sequence analysis indicated that splt97 has 8-50% amino acid identity with the homologues of other known baculoviruses including autographa californica multicapsid nucleopolyhedrovirus (acmnpv) orf108. splt97 transcription in s ...200616525738
the use of a recombinant baculovirus expressing a chitinase from the hard tick haemaphysalis longicornis and its potential application as a bioacaricide for tick control.baculoviruses are specific insect pathogens used as selective biological insecticides on lepidopteran insects. we have tested a recombinant baculovirus expressing a chitinase gene for its efficacy as a tick bioacaricide. the recombinant autographa californica multiple nuclear polyhedrosis virus expressing a chitinase enzyme (acmnpv-cht1) from the hard tick, haemaphysalis longicornis, was constructed and found to have a novel bioacaricidal effect against ticks. the recombinant baculovirus was use ...200616292677
functional expression of lepidopteran-selective neurotoxin in baculovirus: potential for effective pest management.recombinant baculovirus expressing insect-selective neurotoxins derived from venomous animals are considered as an attractive alternative to chemical insecticides for efficient insect control agents. recently we identified and characterized a novel lepidopteran-selective toxin, buthus tamulus insect-selective toxin (butait), having 37 amino acids and eight half cysteine residues from the venom of the south indian red scorpion, mesobuthus tamulus. the synthetic toxin gene containing the butait se ...200616406338
functional analysis of evolutionary conserved clustering of bzip binding sites in the baculovirus homologous regions (hrs) suggests a cooperativity between host and viral transcription factors.the genome of the autographa californica multinucleocapsid polyhedrosis virus (acmnpv) contains nine interspersed homologous regions (hrs) that function as potent enhancer sequences when linked in cis to either viral or heterologous rna polymerase ii-dependent promoters. their activity is strongly increased by the binding of the major immediate early viral transregulator ie1 on 28-mer palindromic sites present in hrs. we show that hrs of acmnpv additionally carry, in the interpalindromic sequenc ...200616198391
characterization of baculovirus autographa californica multiple nuclear polyhedrosis virus infection in mammalian cells.the baculovirus autographa californica multiple nuclear polyhedrosis virus (acmnpv) is used as a vector in many gene therapy studies. wild-type acmnpv infects many mammalian cell types in vitro, but does not replicate. we investigated the dynamics of acmnpv genomic dna in infected mammalian cells and used flow cytometric analysis to demonstrate that recombinant baculovirus containing a cytomegalovirus immediate early promoter/enhancer with green fluorescent protein (gfp) expressed high levels of ...200616545777
a novel method for in vitro radiolabeling and testing enveloped viruses by phosphatidylethanolamine n-methyltransferase and host cell-specific binding.the present study developed a novel virus labeling and testing method, referred to as an envelope-labeled virus assay (elva), in which virus envelope is labeled in vitro by the action of phosphatidylethanolamine n-methyltransferase (pemt) and tested through a host cell-specific binding. a recombinant strain (vgfpuv) of autographa californica multiple nuclear polyhedrosis virus (acmnpv) and spodoptera frugiperda (sf-9) insect cells were used as a model of viruses and host cells, respectively. the ...200616552776
efficient and stable gene expression in rabbit intervertebral disc cells transduced with a recombinant baculovirus vector.a recombinant baculovirus vector expressing gfp (ac-cmv-gfp) was tested in rabbit intervertebral disc cells cultured in monolayer in vitro. direct infection of intervertebral disc cells was then assessed in vivo.200616582845
expression of bioactive recombinant bovine interferon-gamma using baculovirus.the precise role of bovine interferon-gamma (boifn-gamma) in disease and therapy is still poorly defined. clearly it is involved in defence against parasites, bacteria, viruses and possibly tumor cells. this paper reports the expression of boifn-gamma in a baculovirus system to generate a fully functional recombinant protein. bovine interferon-gamma cdna was cloned from mitogen stimulated peripheral blood mononuclear cell (pbmc) rna utilizing the reverse transcription-polymerase chain reaction ( ...200616608121
enhancing insecticidal efficacy of baculovirus by early expressing an insect neurotoxin, lqhit2, in infected trichoplusia ni larvae.lqhit(2), an insect specific neurotoxin from the venom of leiurus quinquestriatus hebraeus, has been demonstrated to improve insecticidal efficacy of autographa californica nuclear polyhedrosis virus (acmnpv). a polyhedrin-positive recombinant acmnpvvacp(hsp70)egfp/p(pag90)it(2) was engineered for larvae to express the enhanced green fluorescence protein (egfp) and lqhit(2) under the control of p(hsp70) and p(pag90) promoters, respectively. this would allow a visual observation of the viral infe ...200616636828
improving baculovirus transduction of mammalian cells by surface display of a rgd-motif.an rgd-containing peptide, comprising 23 amino acids from the foot-and-mouth disease virus (fmdv) vp1 protein was engineered into the envelope of autographa californica nuclear polyhedrosis virus surface (acnpv) using two different display strategies. the rgd-motif is a well-described tripeptide, that by binding to cell surface integrins facilitates virus entry into cells. this epitope was displayed, either by directly modifying the native major envelope protein gp64 of acnpv, or by incorporatin ...200616716431
mapping the conformational epitope of a neutralizing antibody (acv1) directed against the acmnpv gp64 protein.the envelope glycoprotein gp64 of autographa californica nucleopolyhedrovirus (acmnpv) is necessary and sufficient for the acid-induced membrane fusion activity that is required for fusion of the budded virus (bv) envelope and the endosome membrane during virus entry. infectivity of the budded virus (bv) is neutralized by acv1, a monoclonal antibody (mab) directed against gp64. prior studies indicated that acv1 recognizes a conformational epitope and does not inhibit virus attachment to the cell ...200616777166
a recombinant truncated cry1ca protein is toxic to lepidopteran insects and forms large cuboidal crystals in insect cells.a truncated version of the cry1ca gene from bacillus thuringiensis was introduced into the genome of autographa californica multiple nucleopolyhedrovirus (acmnpv) under the control of two promoters. a recombinant virus (vsyncry1c) was isolated and used to infect insect cells in culture and insect larvae. structural and ultrastructural analysis of insects infected with vsyncry1c showed the formation of large cuboidal crystals inside the cytoplasm of insect cells in culture and in insect cadavers ...200616972133
autographa californica multiple nucleopolyhedrovirus nucleocapsid assembly is interrupted upon deletion of the 38k gene.38k (ac98) of autographa californica multiple nucleopolyhedrovirus (acmnpv) is a highly conserved baculovirus gene whose function is unknown. to determine the role of 38k in the baculovirus life cycle, a 38k knockout bacmid containing the acmnpv genome was generated through homologous recombination in escherichia coli. furthermore, a 38k repair bacmid was constructed by transposing the 38k open reading frame with its native promoter region into the polyhedrin locus of the 38k knockout bacmid. af ...200616987976
baculovirus display: a multifunctional technology for gene delivery and eukaryotic library development.for over a decade, phage display has proven to be of immense value, allowing selection of a large variety of genes with novel functions from diverse libraries. however, the folding and modification requirements of complex proteins place a severe constraint on the type of protein that can be successfully displayed using this strategy, a restriction that could be resolved by similarly engineering a eukaryotic virus for display purposes. the quite recently established eukaryotic molecular biology t ...200616997010
genome of the most widely used viral biopesticide: anticarsia gemmatalis multiple nucleopolyhedrovirus.the genome of anticarsia gemmatalis multiple nucleopolyhedrovirus isolate 2d (agmnpv-2d), which is the most extensively used virus pesticide in the world, was completely sequenced and shown to have 132 239 bp (g+c content 44.5 mol%) and to be capable of encoding 152 non-overlapping open reading frames (orfs). three orfs were unique to agmnpv-2d, one of which (ag31) had similarity to eukaryotic poly(adp-ribose) polymerases. the lack of chia and v-cath may explain some of the success and growth of ...200617030857
[a novel application of baculovirus in mammalian gene therapy].baculovirus has been widely used for the production of recombinant proteins in insect cells. the extremely high yield by baculovirus-infected insect cells or larvae makes it an attractive tool for pharmaceutical protein production. since the finding that baculovirus can efficiently transduce mammalian cells, the applications of baculovirus have been greatly expanded. although acmnpv (autographa californica multiple npv) fails to replicate in vertebrate cells, it does express alien genes with lev ...200617037077
response of immunocompetent and immunosuppressed spodoptera littoralis larvae to baculovirus infection.the mediterranean lepidopteran pest spodoptera littoralis is highly resistant to infection with the autographa californica multiple nucleopolyhedrovirus (acmnpv) via the oral route, but highly sensitive to infection with budded virus (bv) via the intrahaemocoelic route. to study the fate of acmnpv infection in s. littoralis, vhsgfp, an acmnpv recombinant that expresses the reporter green fluorescent protein gene under the control of the drosophila heat-shock promoter, and high-resolution fluores ...200616847117
lepidopteran cell lines after long-term culture in alternative media: comparison of growth rates and baculovirus replication.three insect cell lines, iplb-ldfb and iplb-ldeita from gypsy moth fat body and embryos and ufl-ag-286 from velvetbean caterpillar embryos, have been concurrently maintained for 1 to 12 yr on two media formulations, modified tc-100 containing 9% fetal bovine serum and ex-cell 400, a commercial serum-free medium (sfm). cells grown in each medium were tested for susceptibility to and productivity of various multiply embedded nucleopolyhedroviruses. the three lines chosen for these experiments fall ...200616848634
the autographa californica nuclear polyhedrosis virus acnpv induces functional maturation of human monocyte-derived dendritic cells.the initiation of an adaptive immune response is critically dependent on the activation of dendritic cells (dcs). therefore, vaccination strategies targeting dcs have to ensure a proper presentation of the immunogen as well as an activation of dcs to accomplish their full maturation. viral vectors can achieve gene delivery and a subsequent presentation of the expressed immunogen, however, the immunization efficiency may be hampered by an inhibition of dc activation. here we report that the insec ...200616870312
the compatibility of a nucleopolyhedrosis virus control with resistance management for bacillus thuringiensis: co-infection and cross-resistance studies with the diamondback moth, plutella xylostella.the use of genetically modified crops expressing bacillus thuringiensis (bt) toxins can lead to the reduction in application of broad-spectrum pesticides and an increased opportunity for supplementary biological control. bt microbial sprays are also used by organic growers or as part of integrated pest management programs that rely on the use of natural enemies. in both applications the evolution of resistance to bt toxins is a potential problem. natural enemies (pathogens or insects) acting in ...200616905146
temporal transcription program of recombinant autographa californica multiple nucleopolyhedrosis virus.baculoviruses, a family of large, rod-shaped viruses that mainly infect lepidopteran insects, have been widely used to transduce various cells for exogenous gene expression. nonetheless, how a virus controls its transcription program in cells is poorly understood. with a custom-made baculovirus dna microarray, we investigated the recombinant autographa californica multiple nucleopolyhedrosis virus (acmnpv) gene expression program in lepidopteran sf21 cells over the time course of infection. our ...200616940511
group i but not group ii npv induces antiviral effects in mammalian cells.nucleopolyhedrovirus (npv) is divided into group i and group ii based on the phylogenetic analysis. it has been reported that group i npvs such as autographa californica multiple npv (acmnpv) can transduce mammalian cells, while group ii npvs such as helicoverpa armigera single npv (hasnpv) cannot. here we report that acmnpv was capable of stimulating antiviral activity in human hepatoma cells (smmc-7721) manifested by inhibition of vesicular stomatitis virus (vsv) replication. in contrast, the ...200617172054
optimization of baculovirus transduction on freestyle293 cells for the generation of influenza b/lee/40.recombinant baculovirus expression vectors derived from the autographa californica nuclear polyhedrosis virus can serve as efficient gene-transfer vehicles for transient expression of recombinant proteins in a wide range of mammalian cell types and are able to produce multisubunit particles such as viruses or virus like particles. in this study, we constructed eight recombinant baculoviruses each containing one of the influenza b/lee/40 virus genes in a bidirectional expression cassette for simu ...200617172661
an orip/ebna-1-based baculovirus vector with prolonged and enhanced transgene expression.the baculovirus autographa californica multiple nucleopolyhedrovirus (acmnpv) has been explored as a gene delivery vehicle for a variety of mammalian cell lines. however, the transient expression nature due to its incapability to replicate in mammalian cells and insufficient transduction efficiency limit its application.200617051599
functional role of the cytoplasmic tail domain of the major envelope fusion protein of group ii baculoviruses.f proteins from baculovirus nucleopolyhedrovirus (npv) group ii members are the major budded virus (bv) viral envelope fusion proteins. they undergo furin-like proteolysis processing in order to be functional. f proteins from different baculovirus species have a long cytoplasmic tail domain (ctd), ranging from 48 (spodoptera litura multicapsid npv [mnpv]) to 78 (adoxophyes honmai npv) amino acid (aa) residues, with a nonassigned function. this ctd is much longer than the ctd of gp64-like envelop ...200617071930
characterization of antheraea pernyi nucleopolyhedrovirus p11 gene, a homologue of autographa californica nucleopolyhedrovirus orf108.antheraea pernyi nucleopolyhedrovirus (apnpv) p11 gene is 309 bp long, potentially encoding 102 amino acids with a predicted molecular weight of 11.2 kda. apnpv p11 gene was cloned into the prokaryotic expression vector pqe-30 and p11 was expressed in e. coli m15. polyclonal antiserum was made against 6xhis tagged p11 protein expressed in e. coli m15. p11 gene transcription was detected as early as 36 h post-infection (p.i.) in tussah pupa and remain at high level up to 96 h p.i. structural loca ...200717072759
characterization of acmnpv with a deletion of me53 gene.the autographa californica multiple nucleopolyhedrovirus (acmnpv) me53 gene, which was previously reported as one of the major early-transcribed genes, was deleted through homologous recombination from an acmnpv genome propagated as a bacmid dna in e. coli, generating a me53 gene knockout bacmid. green fluorescent protein (gfp) expression analysis and supernatant passage assay revealed that the me53 knockout bacmid was unable to replicate in cell culture, while me53 repair bacmid, which was gene ...200717096186
in vivo gene transfer into the honeybee using a nucleopolyhedrovirus vector.the honeybee apis mellifera l. is a social insect and one of the most industrially important insects. we examined whether a baculovirus-mediated retrotransposon is applicable to in vivo transfer of exogenous genes to the honeybees. honeybee larvae and pupae were injected with two types of recombinant autographa californica nucleopolyhedrovirus (acnpv) vectors, one that includes the enhanced green fluorescent protein gene (egfp) as a reporter to be inserted into the honeybee genome, and another t ...200717125735
p13 of leucania separata multiple nuclear polyhedrosis virus affected the polyhedra and budded virions yields of acmnpv.p13 gene was first described by our laboratory in leucania separata multiple nuclear polyhedrovirus (ls-p13, orf114) back to 1995. however, the functions of ls-p13 and its reported homologues remained unknown. in order to probe the function of ls-p13, recombinant autographa californica nucleopolyhedroviruses (racmnpvs) were constructed to express ls-p13 in the sf9 cells at early, late or early/late phase. observations of microscope showed that the expression of ls-p13 could decrease the yield of ...200717141348
the homingbac baculovirus cloning system: an alternative way to introduce foreign dna into baculovirus genomes.an in vitro baculovirus cloning system has been developed for direct cloning of foreign dna into baculovirus genomes. this system is called the "homingbac system" because it uses homing endonucleases. the homingbac system was engineered into the baculoviruses acmnpv, bmnpv, pxmnpv, romnpv, hasnpv and hzsnpv. all homingbac viruses were designed to retain the polyhedra phenotype so that they could be inoculated per os to insects. this is the first time a common in vitro baculovirus cloning system ...200717141883
mutational analysis of active site residues of chitinase from bombyx mori nucleopolyhedrovirus.infection of bombyx mori larvae with b. mori nucleopolyhedrovirus (bmnpv) results in liquefaction of the host. this process is attributed to the synergistic action of two virus-encoded genes, chitinase (v-chia) and cathepsin (v-cath). previous studies have suggested that autographa californica nucleopolyhedrovirus (acmnpv) cath cannot be processed within infected cells in the absence of acmnpv chia. to investigate the interactions between v-chia and v-cath, we generated a recombinant bmnpv (103c ...200717145091
bv/odv-e26: a palmitoylated, multifunctional structural protein of autographa californica nucleopolyhedrovirus.autographa californica nucleopolyhedrovirus ac16 is 1 of 17 genes conserved within type 1 nucleopolyhedroviruses. this report demonstrates that multiple isoforms of the protein encoded by ac16, bv/odv-e26 (e26), are present in the infected cell. one form of e26 associates with viral dna or dna-binding proteins, while a second form associates with intracellular membranes and this is likely due to palmitoylation. the different forms of e26 present unique epitopes that can be discriminated by antis ...200717169392
prediction of recombinant protein production in an insect cell-baculovirus system using a flow cytometric technique.the baculovirus expression vector system (bevs) utilising the autographa californica nucleopolyhedrovirus (acmnpv) is widely becoming the system of choice for the production of many recombinant protein products due to the high yields obtained. however, there is a need to develop a simple reliable on-line method to monitor the production of recombinant proteins that have no intrinsic reporter properties. here we utilise flow cytometry to measure cell size, granularity and dna content in a single ...200717643445
biochemical characterization of sf9 sp-family-like protein factors reveals interesting features.we earlier documented the involvement of novel sp-family-like protein factors in transcription from the autographa californica nucleopolyhedrovirus (acnpv) polyhedrin (polh) gene promoter [ramachandran et al. (2001) j. biol. chem. 276: 23440-23449]. these zinc-dependent sp-like factors bind to two putative sp-factor-binding motifs, present within the acsp sequence upstream of the polh promoter, with very high affinity (k(d) = 2.1 x 10(-12) m). like other polh-promoter-associated host transcripti ...200717653621
purification and characterization of a recombinant rat prohaptoglobin expressed in baculovirus-infected sf9 insect cells.to generate hemoglobin-free full-length haptoglobin the cdna encoding rat haptoglobin alphabeta subunits was cloned into shuttle vector pvt-bac-his and used to produce a recombinant baculovirus autographa californica nuclear polyhedrosis virus (acnpv) as an expression vector, named hpacnpv. recombinant virus was used to infect spodoptera frugiperda (sf9) insect cells. the 50 kda protein expressed was mostly secreted into the culture medium at relatively high titer (15 microg/ml) and was found to ...200717681809
reprogramming the chia expression profile of autographa californica multiple nucleopolyhedrovirus.expression of chia and v-cath rna and enzyme activity in wild-type autographa californica multiple nucleopolyhedrovirus (acmnpv) was compared with that of recombinant acmnpv viruses reprogrammed for expression of the endogenous chia. to establish a baseline for our recombinant acmnpv studies, we compared, for the first time, the temporal expression profiles of both acmnpv chia transcription and translation simultaneously. the rate of intracellular chitinase accumulation during acmnpv infection f ...200717698657
development and characterization of a continuous cell line, afkm-on-h, from hemocytes of the european corn borer ostrinia nubilalis (hübner) (lepidoptera, pyralidae).the corn borer, ostrinia nubilalis, is a very important pest in different countries, and the in vitro system of the insect could be a useful tool for isolation and characterization of the pathogens and physiological responses of the insect. in this context, a cell line was derived from the hemocytes of the european corn borer and was named afkm-on-h for, respectively, o. nubilalis, armand frappier, king mongkut institutes, and hemocytes. this cell line was initiated and maintained in ex-cell 400 ...200717846857
acmnpv orf38 protein has the activity of adp-ribose pyrophosphatase and is important for virus replication.the orf38 of autographa californica multiple nucleopolyhedrovirus (acmnpv), or acorf38, contains a conserved motif of nudix (nucleotide diphosphate x) superfamily. it has the highest homology with adp-ribose pyrophosphatase (adprase), a subfamily of nudix pyrophosphatase. in the current study, recombinant acorf38 protein was prepared and shown to have adprase activity, with a km of 204 microm, and k(cat) of 6.96 s(-1) at ph 8.0 and 5 mm mgcl2. the transcription of acorf38 was detected 2 h postin ...200717174373
proteins associated with culex nigripalpus nucleopolyhedrovirus occluded virions.occlusion-derived virions (odvs) of the nucleopolyhedrovirus of culex nigripalpus (cuninpv) were purified by ludox density gradient ultracentrifugation, and the proteins were separated by one-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis. proteins were identified by using edman sequencing, matrix-assisted laser desorption ionization-time of flight mass spectrometry, nanoelectrospray quadrupole time-of-flight mass spectrometry, or a combination of these methods. half of th ...200717301145
increased plasma selenium levels correlate with elevated resistance of heliothis virescens larvae against baculovirus infection.we reported that dietary selenium (se) impacted the growth and development of trichoplusia ni reared for many generations on diet containing extremely low levels of se. larvae had an elevated resistance to per os infection with a baculovirus. in this study, we examine how dietary se (in the form of selenite) affects the growth, development, and se content of heliothis virescens that have been laboratory reared for less than two years. larvae fed a commercial tobacco budworm diet supplemented wit ...200717316679
function and oligomerization study of the leucine zipper-like domain in p13 from leucania separata multiple nuclear polyhedrosis virus.the p13 gene is uniquely present in group ii nucleopolyhedroviruses (npvs) and some granuloviruses, but not in group i npvs. p13 gene was first described by our laboratory in leucania separatamultiple nuclear polyhedrosis virus (ls-p13) in 1995. however, the functions of ls-p13 and of its homologues are unknown. when ls-p13 was inserted into autographa californica nucleopolyhedrovirus, a group i npv, polyhedra yield was inhibited. however, this inhibition was prevented when the leucine zipper-li ...200717394774
characterization of a baculovirus lacking the dbp (dna-binding protein) gene.autographa californica multiple nucleopolyhedrovirus (acmnpv) encodes two proteins that possess properties typical of single-stranded dna-binding proteins (ssbs), late expression factor-3 (lef-3), and a protein referred to as dna-binding protein (dbp). whereas lef-3 is a multi-functional protein essential for viral dna replication, transporting helicase into the nucleus, and forms a stable complex with the baculovirus alkaline nuclease, the role for dbp in baculovirus replication remains unclear ...200717449080
the autographa californica m nucleopolyhedrovirus fibroblast growth factor accelerates host mortality.the fibroblast growth factor (vfgf) gene encoded by autographa californica m nucleopolyhedrovirus (acmnpv) has been shown to share functional properties with cellular fgfs; it is a secreted protein, binds heparin, and stimulates motility of insect cells. we previously reported that viruses containing or lacking vfgf produced similar yields of budded virus and had similar kinetics of viral dna and protein syntheses in cultured cells. in this study, we characterized these viruses in two permissive ...200717459443
the acmnpv pp31 gene is not essential for productive acmnpv replication or late gene transcription but appears to increase levels of most viral transcripts.the pp31 gene of autographa californica multicapsid nucleopolyhedrovirus (acmnpv) encodes a phosphorylated dna binding protein that associates with virogenic stroma in the nuclei of infected cells. prior studies of pp31 by transient late expression assays suggested that pp31 may play an important role in transcription of acmnpv late genes [todd, j. w., passarelli, a. l., and miller, l. k. (1995). eighteen baculovirus genes, including lef-11, p35, 39k, and p47, support late gene expression. j. vi ...200717467768
the heptad repeats region is essential for acmnpv p10 filament formation and not the proline-rich or the c-terminus basic regions.baculovirus p10 protein is a small conserved protein and is expressed as bundles of filaments in the host cell during the late phase of virus infection. so far the published results on the domain responsible for filament structural formation have been contradictory. electron microscopy revealed that the c-terminus basic region was involved in filament structural formation in the autographa californica multiple nucleocapsid nucleopolyhedrovirus (acmnpv) [van oers, m.m., flipsen, j.t., reusken, c. ...200717481692
ac18 is not essential for the propagation of autographa californica multiple nucleopolyhedrovirus.orf18 (ac18) of autographa californica multiple nucleopolyhedrovirus (acmnpv) is a highly conserved gene in lepidopteran nucleopolyhedroviruses, but its function remains unknown. in this study, an ac18 knockout acmnpv bacmid was generated to determine the role of ac18 in baculovirus life cycle. after transfection of sf-9 cells, the ac18-null mutant showed similar infection pattern to the parent virus and the ac18 repair virus with respect to the production of infectious budded virus, occlusion b ...200717573091
the n-terminal hydrophobic sequence of autographa californica nucleopolyhedrovirus pif-3 is essential for oral infection.the autographa californica nucleopolyhedrovirus (acmnpv) open reading frame 115 has been identified as a per os infection factor (pif-3) and is essential for oral infection. here, we have characterized the pif-3 of acmnpv in more detail. the pif-3 transcripts were detected from 12 to 96 h post-infection (hpi) in sf9 cells infected with acmnpv. polyclonal antiserum first recognized a 25-kda protein at 36 hpi. western blot analysis indicated that pif-3 is a component of occlusion-derived virus but ...200717585368
characterization of baculovirus constructs lacking either the ac 101, ac 142, or the ac 144 open reading frame.to investigate the role of the gene products encoded from the open reading frames 101, 142, and 144 of autographa californica multiple nucleopolyhedrovirus (acmnpv), a set of bacmid knockout and repair constructs were generated. the repair genes were engineered to contain an ha epitope tag at their c-termini. the results of transfection-infection assays and growth curve analyses showed that the ac 101, 142, and 144 genes were required for infectious virus production. to better characterize the r ...200717585983
biosafety of recombinant and wild type nucleopolyhedroviruses as bioinsecticides.the entomopathogenic autographa californica (speyer) nucleopolyhedrovirus (acmnpv) has been genetically modified to increase its speed of kill. the potential adverse effects of a recombinant acmnpv (acaait) as well as wild type acmnpv and wild type spodoptera littoralis npv (slnpv) were studied. cotton plants were treated with these viruses at concentrations that were adjusted to resemble the recommended field application rate (4 x 10(12) pibs/feddan, feddan = 4,200 m2) and 3rd instar larvae of ...200717617674
stimulation of baculovirus transcriptome expression in mammalian cells by baculoviral transcriptional activators.autographa californica multiple nucleopolyhedrovirus (acmnpv), the type species of the family baculoviridae, is an insect-specific virus that can enter a variety of mammalian cells. the potential of this versatile virus for protein expression or gene therapy in mammalian cells has become the focus of many studies. in most mammalian cells, transduced acmnpv genes are either not expressed or expressed at an extremely low level. here, we studied the effects of the two major acmnpv trans-activators, ...200717622620
autographa californica multiple nucleopolyhedrovirus exon0 (orf141) is required for efficient egress of nucleocapsids from the nucleus.autographa californica multiple nucleopolyhedrovirus (acmnpv) exon0 (orf141) has been shown to be required for the efficient production of budded virus (bv). the deletion of exon0 reduces the level of bv production by up to 99% (x. dai, t. m. stewart, j. a. pathakamuri, q. li, and d. a. theilmann, j. virol. 78:9633-9644, 2004); however, the function or mechanism by which exon0 affects bv production is unknown. in this study, we further elucidated the function of exon0 by investigating the locali ...200717626083
tissue specificity of a baculovirus-expressed, basement membrane-degrading protease in larvae of heliothis virescens.scathl is a cathepsin l-like cysteine protease from the flesh fly, sarcophaga peregrina, which digests components of the basement membrane during insect metamorphosis. a recombinant baculovirus (acmlf9.scathl) expressing scathl kills larvae of the tobacco budworm heliothis virescens significantly faster than the wild type virus and triggers melanization and tissue fragmentation shortly before death. the tissue fragmentation was assumed to be a direct consequence of basement membrane degradation ...200717959212
molecular biology of the baculovirus occlusion-derived virus envelope.study of the biology of the occlusion-derived virus (odv) of the baculovirus autographa californica nucleopolyhedrovirus provides opportunities to reveal new discoveries into the mechanism of several cellular pathways. the synchronous pulse of multiple odv envelope proteins that integrate into the endoplasmic reticulum (er) and traffic to the nuclear membranes on their way to the odv envelope provide a unique tool to study the mechanisms of integral membrane protein trafficking from the er to th ...200717979668
baculovirus envelope fusion proteins f and gp64 exploit distinct receptors to gain entry into cultured insect cells.group ii nucleopolyhedroviruses (npvs), e.g. helicoverpa armigera (hear) npv and spodoptera exigua (se) mnpv (multiple npv), lack a gp64-like protein that is present in group i npvs, e.g. autographa californica (ac)mnpv, but have an unrelated envelope fusion protein named f. three acmnpv viruses were constructed by introducing acmnpv gp64, hearnpv f or semnpv f genes, respectively, into a gp64-negative acmnpv bacmid. sf21 cells were incubated with different amounts of inactivated budded virus to ...200718024899
a baculovirus alkaline nuclease knockout construct produces fragmented dna and aberrant capsids.dna replication of bacmid-derived constructs of the autographa californica multiple nucleocapsid nucleopolyhedrovirus (acmnpv) was analyzed by field inversion gel electrophoresis (fige) in combination with digestion at a unique eco81i restriction enzyme site. three constructs were characterized: a parental bacmid, a bacmid deleted for the alkaline nuclease gene, and a bacmid from which the gp64 gene had been deleted. the latter was employed as a control for comparison with the alkaline nuclease ...200717046043
using cell size kinetics to determine optimal harvest time for spodoptera frugiperda and trichoplusia ni bti-tn-5b1-4 cells infected with a baculovirus expression vector system expressing enhanced green fluorescent protein.infecting insect cells with a baculovirus expression vector system (bevs) is an increasingly popular method for the production of recombinant proteins. due to the lytic nature of the system, however, determining the optimal harvest time is critical for maximizing protein yield. we found that measuring the change in average diameter during the progress of infection with an automated cell analysis system (cedex hires, innovatis ag) could be used to determine the time of maximum protein production ...200719003016
highly passage of spodoptera litura cell line causes its permissiveness to baculovirus infection.it is well known that the characteristics of cell lines possibly alter when cell lines are at high-passage number because of the environmental selection. we do not know whether non-permissive or low-permissive cell lines could become permissive or more permissive to virus infection after over-high passage. in the present studies, the alteration of the permissiveness of spodoptera litura cell line sl-zsu-1 to three baculovirus infection was investigated after over-high passage, and the possible m ...200819003180
a bacmid approach to the genetic manipulation of granuloviruses.a cydia pomonella granulovirus (cpgv) bacmid has been constructed, which allows rapid and efficient production of recombinant baculoviruses in escherichia coli. an 8.6kbp bacterial dna cassette derived from the acmnpv bac-to-bac system was ligated into a unique paci restriction site within an intergenic region flanking the dna ligase gene of the cpgv genome. the cpgv bacmids produced in e. coli were transfected into a cpgv-permissive c. pomonella cell line and the transfected cells fed to larvae ...200818602169
analysis of inactivation of acmnpv under various conditions by the elva method.inactivation of baculoviruses was examined under various conditions by elva, which enzymatically labels the virus envelope with radioisotopes and detects the labeled viruses by host cell-specific binding. when baculoviruses were incubated in a 50-ml culture tube, they rapidly lost infective ability by more than 75% of the initial titer in 2 h even at an insect cell cultivation temperature of 27 degrees c. altering ph from neutral significantly decreased the virus titer. repetition of freezing an ...200818603767
effects of temperature and shear force on infectivity of the baculovirus autographa californica m nucleopolyhedrovirus.virus stability and infectivity during stressful conditions was assessed to establish guidelines for future virus filtration experiments and to contribute to the body of knowledge on a widely used virus. a recombinant baculovirus of autographa californica m nucleopolyhedrovirus (acmnpv), vhsgfp, was incubated at 15-65 degrees c. a 2-log decrease in virus infectivity occurred after virus incubation above 45 degrees c. the activation energy of virus deactivation was circa 108 kj/mol. dynamic light ...200818760306
identification of baculoviral factors required for the activation of enhancer-like polyhedrin upstream (pu) sequence.previously, we identified a novel enhancer-like element, the polyhedrin upstream (pu) sequence, in the genome of the baculovirus autographa californica multiple nucleopolyhedrovirus (acmnpv), which activates several early promoters. the activation requires co-infection of acmnpv, suggesting that viral gene products are needed for pu-mediated promoter activation. dna replication assay showed that the pu sequence did not assist in dna replication and suggested its involvement in activated transcri ...200818760315
trypsin cleavage of the baculovirus occlusion-derived virus attachment protein p74 is prerequisite in per os infection.baculovirus occlusion-derived virions (odvs) contain a number of infectivity factors essential for the initiation of infection in larval midgut cells. deletion of any of these factors neutralizes infectivity by the per os route. we have observed that p74 of the group i alphabaculovirus autographa californica multiple nucleopolyhedrovirus (acmnpv) is n-terminally cleaved when a soluble form of the protein was incubated with insect midgut tissues under alkaline conditions and that cleavage was pre ...200818796706
characterization of a bombyx mori nucleopolyhedrovirus with bmvp80 disruption.a bmnpv bacmid with the bmvp80 gene disrupted was constructed using the et-recombination system in escherichia coli to investigate the role of bmvp80 during the baculovirus life cycle. disruption of bmvp80 resulted in single cell infection phenotype, whereas a rescue bmbacmid restored budded virus titers to wild type levels; however, the homologous gene ac104 (acvp80) from acmnpv could not complement the bmbacmid lacking a functional bmvp80 gene. electron microscopy of cells transfected with bmn ...200818809445
development of a novel preparation method of recombinant proteoliposomes using baculovirus gene expression systems.we have developed a novel method for the preparation of 'recombinant proteoliposomes'. membrane proteins were expressed on budded virus (bv) envelopes using baculovirus gene expression systems, and proteoliposomes were prepared by fusion of these viruses with liposomes. first, plasmid dna containing the gene for the thyroid-stimulating hormone receptor (tshr) or the acetylcholine receptor alpha-subunit (achralpha) was co-transfected with wild type virus [autographa californica nuclear polyhedros ...200818836200
autographa californica multiple nucleopolyhedrovirus ac53 plays a role in nucleocapsid assembly.autographa californica multiple nucleopolyhedrovirus (acmnpv) orf53 (ac53) is a highly conserved gene existing in all sequenced lepidoptera and hymenoptera baculoviruses, but its function remains unknown. to investigate its role in the baculovirus life cycle, an ac53 deletion virus (vac(ac53ko-ph-gfp)) was generated through homologous recombination in escherichia coli. fluorescence and light microscopy and titration analysis revealed that vac(ac53ko-ph-gfp) could not produce infectious budded vi ...200818851866
autographa californica multiple nucleopolyhedrovirus 38k is a novel nucleocapsid protein that interacts with vp1054, vp39, vp80, and itself.it has been shown that the autographa californica multiple nucleopolyhedrovirus (acmnpv) 38k (ac98) is required for nucleocapsid assembly. however, the exact role of 38k in nucleocapsid assembly remains unknown. in the present study, we investigated the relationship between 38k and the nucleocapsid. western blotting using polyclonal antibodies raised against 38k revealed that 38k was expressed in the late phase of infection in acmnpv-infected spodoptera frugiperda cells and copurified with budde ...200818922869
mechanism analysis of broad-spectrum disease resistance induced by expression of anti-apoptotic p35 gene in tobacco.studies have shown that transgenic plants expressing antiapoptotic genes from baculovirus and animals increase resistance to biotic and abiotic stress. however, the mechanism under these resistances is conjectural, or in some cases even controversy. in the present study, the p35 gene from baculovirus autographa californica multicapsid nucleopolyhedrovirus (acmnpv) was expressed in tobacco, and for the first time p35 protein was detected in transgenic plants by western blotting. inoculation of t1 ...200819149181
isolation and characterization of a baculovirus associated with the insect parasitoid wasp, cotesia marginiventris, or its host, trichoplusia ni.a multiple nucleopolyhedrovirus (mnpv) was isolated from trichoplusia ni (hübner) (lepidoptera: noctuidae) larvae that had been stung by the parasitoid cotesia marginiventris (cresson) (hymenoptera: braconidae). the wild type virus was plaque purified by infecting a heliothis subflexa (bcirl- hsam1) cell line and isolating several clones. the mean estimated genomic size of this virus based on psti, bsteii, styi, hindiii restriction profiles was estimated to be 106 +/- 2.5 kbp (mean+/-se). a clon ...200820334593
recombinant expression of garlic virus c (garv-c) capsid protein in insect cells and its potential for the production of specific antibodies.garlic cultivars in brazil are infected by a complex of viruses and for some virus species, such as the allexivirus, purification of the virions is sometimes cumbersume. to overcome this problem, recombinant expression of viral proteins in heterologous systems is an alternative method for producing antibodies. the capsid gene from garlic virus c (garv-c), an allexivirus, was inserted into the genome of autographa californica multiple nucleopolyhedrovirus (acmnpv) generating the recombinant virus ...200816890415
baculovirus-mediated immediate-early gene expression and nuclear reorganization in human cells.baculovirus, autographa californica multiple nucleopolyhedrovirus (acmnpv), has the ability to transduce mammalian cell lines without replication. the general objective of this study was to detect the transcription and expression of viral immediate-early genes in human cells and to examine the interactions between viral components and subnuclear structures. viral capsids were seen in large, discrete foci in nuclei of both dividing and non-dividing human cells. concurrently, the transcription of ...200818042259
autographa californica multiple nucleopolyhedrovirus ac142, a core gene that is essential for bv production and odv envelopment.autographa californica multiple nucleopolyhedrovirus (acmnpv) ac142 is a baculovirus core gene and encodes a protein previously shown to associate with occlusion-derived virus (odv). to determine its role in the baculovirus life cycle, we used the acmnpv bacmid system to generate an ac142 deletion virus (acbac(ac142ko-ph-gfp)). fluorescence and light microscopy revealed that acbac(ac142ko-ph-gfp) exhibits a single-cell infection phenotype. titration assays and western blot confirmed that acbac(a ...200818045640
induction of antitumor acquired immunity by baculovirus autographa californica multiple nuclear polyhedrosis virus infection in mice.the baculovirus autographa californica multiple nuclear polyhedrosis virus (acmnpv) has been studied as a gene therapy vector. here, we demonstrated that acmnpv induces antitumor acquired immunity. these results suggest that acmnpv has the potential to be an efficient virus or tumor therapy agent which induces innate and acquired immunity.200818057182
induction of natural killer cell-dependent antitumor immunity by the autographa californica multiple nuclear polyhedrosis virus.wild-type autographa californica multiple nuclear polyhedrosis virus (acmnpv) infects a variety of mammalian cell types in vitro, but does not replicate in these cells. we investigated the effects of acmnpv in the induction of the immune response and tumor metastasis in mice. after intravenous injection, acmnpv was taken up by the liver and spleen, and preferentially infected dendritic cells (dcs) and b cells in the spleen; costimulatory molecules cd40, cd80, and cd86 were upregulated in the dcs ...200818059370
conserved leucines in n-terminal heptad repeat hr1 of envelope fusion protein f of group ii nucleopolyhedroviruses are important for correct processing and essential for fusogenicity.the heptad repeat (hr), a conserved structural motif of class i viral fusion proteins, is responsible for the formation of a six-helix bundle structure during the envelope fusion process. the insect baculovirus f protein is a newly found budded virus envelope fusion protein which possesses common features to class i fusion proteins, such as proteolytic cleavage and the presence of an n-terminal open fusion peptide and multiple hr domains on the transmembrane subunit f(1). similar to many vertebr ...200818094156
expression of two heterologous proteins depends on the mode of expression: comparison of in vivo and in vitro methods.the yield of two proteins, avidin and green fluorescent protein (gfp), expressed from a modified autographa californica nucleopolyhedrovirus (acmnpv), was compared in sf9 cell culture monolayer, sf21 cell suspension culture and intact spodoptera litura larvae. gfp expressed from the p10 promoter yielded up to 1.5% of total soluble protein in larvae, 20-fold higher than that in monolayer suspension culture. avidin, expressed from the polh promoter, yielded up to 2.3% of total soluble protein in l ...200818175154
gp64 of group i nucleopolyhedroviruses cannot readily rescue infectivity of group ii f-null nucleopolyhedroviruses.the genus nucleopolyhedrovirus (npv) of the family baculoviridae can be subdivided phylogenetically into two groups. the same division can be made on the basis of their budded virus (bv) envelope fusion protein. group i npvs are characterized by the presence of a gp64-like major envelope fusion protein, which is involved in viral attachment and the fusion of virus and cell membrane, and is required for budding of progeny nucleocapsids. group ii npvs have an envelope fusion protein unrelated to g ...200818198373
characterization of the gene delivery properties of baculoviral-based virosomal vectors.the current study reports the production of baculoviral-virosomal vectors consisting of lipoplexes and of the viral glycoprotein (gp64) of baculovirus autographa californica multiple nucleopolyhdrovirus (acmnpv). this study demonstrates that such complexes have an increased transfection capability in a number of cells, including undifferentiated h9 human embryonic stem h9hes cells compared to lipoplexes alone. the gp64-mediated enhancement of gene transfer of lipoplexes is inhibited by the addit ...200818207578
characterization of an egyptian spodoptera littoralis nucleopolyhedrovirus and a possible use of a highly conserved region from polyhedrin gene for nucleopolyhedrovirus detection.an egyptian isolate of spodoptera littoralis nucleopolyhedrovirus (splinpv) was tested for its potential as biocontrol agent in comparison to autographa californica multiple nucleopolyhedrovirus (acmnpv). comparative assays of splinpv and acmnpv against 2nd instar larvae of spodoptera littoralis revealed 4-fold greater susceptibility of s. littoralis to acmnpv than to splinpv based on lc50 values for the two viruses. the lt50s determined for splinpv and acmnpv using lc50 of the virus against 2nd ...200818215282
functional analysis of the transmembrane (tm) domain of the autographa californica multicapsid nucleopolyhedrovirus gp64 protein: substitution of heterologous tm domains.gp64, the major envelope glycoprotein of the autographa californica multicapsid nucleopolyhedrovirus (acmnpv) budded virion, is important for host cell receptor binding and mediates low-ph-triggered membrane fusion during entry by endocytosis. in the current study, we examined the functional role of the acmnpv gp64 transmembrane (tm) domain by replacing the 23-amino-acid gp64 tm domain with corresponding tm domain sequences from a range of viral and cellular type i membrane proteins, including o ...200818216100
autographa californica multiple nucleopolyhedrovirus ac66 is required for the efficient egress of nucleocapsids from the nucleus, general synthesis of preoccluded virions and occlusion body formation.although orf66 (ac66) of autographa californica multiple nucleopolyhedrovirus (acmnpv) is conserved in all sequenced lepidopteran baculovirus genomes, its function is not known. this paper describes generation of an ac66 knockout acmnpv bacmid mutant and analyses of the influence of ac66 deletion on the virus replication in sf-9 cells so as to determine the role of ac66 in the viral life cycle. results indicated that budded virus (bv) yields were reduced over 99% in ac66-null mutant infected cel ...200818241908
extended budded virus formation and induction of apoptosis by an acmnpv fp-25/p35 double mutant in trichoplusia ni cells.an autographa californica nucleopolyhedrovirus (acmnpv) mutant (acdefrt) isolated from virus-infected trichoplusia ni (tn-368) cells produced plasma membrane blebbing and caspase-3-like activity late in infection. it also synthesized less polyhedra, but displayed enhanced budded virus formation in tn-368 cells. this phenotype resulted from dual mutations in p35 and fp-25. in this study we showed that enhanced budded virus production occurs because the hourly rate of release of virus from acdefrt ...200818261819
display of heterologous proteins on gp64null baculovirus virions and enhanced budding mediated by a vesicular stomatitis virus g-stem construct.the autographa californica multicapsid nucleopolyhedrovirus (acmnpv) gp64 envelope glycoprotein is essential for virus entry and plays an important role in virion budding. an acmnpv construct that contains a deletion of the gp64 gene is unable to propagate infection from cell to cell, and this defect results from both a severe reduction in the production of budded virions and the absence of gp64 on virions. in the current study, we examined gp64 proteins containing n- and c-terminal truncations ...200817989172
the baculovirus p10 protein of autographa californica nucleopolyhedrovirus forms two distinct cytoskeletal-like structures and associates with polyhedral occlusion bodies during infection.the role of the microtubule-associated p10 protein of baculoviruses is not yet understood. p10 has previously been linked with the formation of a number of cytoskeletal-like or cytoskeleton-associated structures in the nucleus and cytoplasm, thought to be involved in the morphogenesis of virus polyhedral occlusion bodies. the formation of these structures was studied by immunofluorescence laser scanning confocal microscopy in tn368 cells, a model system amenable to the study of virus interaction ...200817991504
proteomics computational analyses suggest that baculovirus gp64 superfamily proteins are class iii penetrenes.members of the baculoviridae encode two types of proteins that mediate virus:cell membrane fusion and penetration into the host cell. alignments of primary amino acid sequences indicate that baculovirus fusion proteins of group i nucleopolyhedroviruses (npv) form the gp64 superfamily. the structure of these viral penetrenes has not been determined. the gp64 superfamily includes the glycoprotein (gp) encoded by members of the thogotovirus genus of the orthomyxoviridae. the entry proteins of other ...200818282283
identification of acmnpv exon0 (ac141) domains required for efficient production of budded virus, dimerization and association with bv/odv-c42 and fp25.the autographa californica multiple nucleopolyhedrovirus (acmnpv) late gene exon0 (ac141) is required for the efficient production of budded virus (bv). exon0 interacts with nucleopcapsid protein bv/odv-c42 and fp25 and enables egress of nucleocapsids from the nucleus to the cytoplasm. this study examines the functional domains of exon0 that play a role in bv production. six putative domains of the 261 amino acid exon0 were deleted and examined for functionality by determining their ability to r ...200818313716
acmnpv ac143 (odv-e18) is essential for mediating budded virus production and is the 30th baculovirus core gene.autographa californica multiple nucleopolyhedrovirus (acmnpv) ac143 (odv-e18) is a late gene that encodes for a predicted 9.6 kda structural protein that locates to the occlusion derived viral envelope and viral induced intranuclear microvesicles [braunagel, s.c., he, h., ramamurthy, p., and summers, m.d. (1996). transcription, translation, and cellular localization of three autographa californica nuclear polyhedrosis virus structural proteins: odv-e18, odv-e35, and odv-ec27. virology 222, 100-1 ...200818328526
inhibition of autographa californica nucleopolyhedrovirus (acnpv) polyhedrin gene expression by dnazyme knockout of its serine/threonine kinase (pk1) gene.dnazyme is known to selectively cleave rna at predetermined site. transfection of autographa californica nucleopolyhedrovirus (acnpv) infected sf9 cells with serine/threonine kinase (pk1) mrna specific dnazymes, dz1 and dz2 to cleave the viral coded (pk1) mrna in between 87th and 88th, and 250th and 251st nucleotide, respectively inhibited the pk1 mrna and its protein expressions. interestingly, polh mrna and protein expressions were also inhibited by these dnazymes despite their inability to cl ...200818353480
baculovirus-mediated interferon alleviates dimethylnitrosamine-induced liver cirrhosis symptoms in a murine model.the wild-type baculovirus autographa californica multiple nuclear polyhedrosis virus (acmnpv) infects a range of mammalian cell types in vitro but does not replicate in these cells. the current study investigated the in vivo effect of acmnpv in the mouse model of liver cirrhosis induced by the mutagen dimethylnitrosamine. intraperitoneal injection of acmnpv induced an immune response. the baculovirus was taken up by the liver and spleen where it suppressed liver injury and fibrosis through the i ...200818369328
plant-mediated alteration of the peritrophic matrix and baculovirus infection in lepidopteran larvae.the peritrophic matrix (pm) lines the midgut of most insects, providing protection to the midgut epithelial cells while permitting passage of nutrients and water. herein, we provide evidence that plant-mediated alteration of the pm contributes to the well-documented inhibition of fatal infection by autographa californica multiple nucleopolyhedrovirus (acmnpv) of heliothis virescens f. larvae fed cotton foliage. we examined the impact of the pm on pathogenesis using a viral construct expressing a ...200818374352
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