| 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 ... | 2008 | 18760315 |
| 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 ... | 2008 | 18796706 |
| 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 ... | 2008 | 18809445 |
| 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 ... | 2008 | 18836200 |
| 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 ... | 2008 | 18851866 |
| 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 ... | 2008 | 18922869 |
| transactivator ie1 is required for baculovirus early replication events that trigger apoptosis in permissive and nonpermissive cells. | immediate early viral protein ie1 is a potent transcriptional activator encoded by baculoviruses. although the requirement of ie1 for multiplication of autographa californica multicapsid nucleopolyhedrovirus (acmnpv) is well established, the functional roles of ie1 during infection are unclear. here, we used rna interference to ablate ie1, plus its splice variant ie0, and thereby define in vivo activities of these early proteins, including gene-specific regulation and induction of host cell apop ... | 2009 | 18945761 |
| 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 ... | 2007 | 19003016 |
| 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 ... | 2008 | 19003180 |
| recombinant protein production by the baculovirus-insect cell system in basal media without serum supplementation. | the production of beta-galactosidase by sf9 cells infected with recombinant autographa californica nucleopolyhedrovirus (acnpv) was investigated in shake-flask culture using two serum-free basal media: grace's medium and tnm-fh (grace's medium supplemented with lactalbumin hydrolysate and yeast extract). at the time of infection, cells grown in serum-supplemented tnm-fh were transferred into fresh basal media without adaptation. the absence of serum depressed the beta-galactosidase yield conside ... | 2003 | 19003201 |
| baculovirus data suggest a common but multifaceted pathway for sorting proteins to the inner nuclear membrane. | multiple unique protein markers sorted to the inner nuclear membrane (inm) from the autographa californica nucleopolyhedrovirus occlusion-derived virus (odv) envelope were used to decipher common elements of the sorting pathway of integral membrane proteins from their site of insertion into the membrane of the endoplasmic reticulum (er) through their transit to the inm. the data show that during viral infection, the viral protein fp25k is a partner for all known odv envelope proteins and that bv ... | 2009 | 19019955 |
| characterization of a new autographa californica multiple nucleopolyhedrovirus (acmnpv) polyhedra mutant. | in the very late phase of baculovirus infection, virions are occluded in a crystalline matrix called polyhedra, which is mainly composed of polyhedrin. this protein is highly conserved among baculoviruses and changes in its amino acid sequence may lead to mutant polyhedra. during the purification of an acmnpv recombinant virus, a mutant virus was isolated. structural and ultrastrutural analysis by light and transmission electron microscopy (tem) of insect cells infected with this mutant virus di ... | 2009 | 19038296 |
| establishment of tetracycline-inducible gene expression systems in the silkworm, bombyx mori. | tetracycline-inducible gene expression (tet-on) system has become one of the first choices for the control of transgenes expression in mammal and drosophila. however, the tet-on systems that have been established in mammalian system or tuned into drosophila do not function in the silkworm, bombyx mori. to construct a functional tet-on system in b. mori, we modified rtta by introducing a transcription activation domain of immediate-early gene 1 of autographa californica nuclear polyhedrosis virus ... | 2009 | 19066730 |
| transgenesis approaches for functional analysis of peptidergic cells in the silkworm bombyx mori. | the domestic silkworm, bombyx mori represents an insect model of great scientific and economic importance. besides the establishment of a stable germline transformation using the piggybac vector, technically feasible methods for in vivo gene delivery and transient gene expression were developed using viral based vectors, especially sindbis viruses and baculoviruses. the recombinant baculovirus, autographa californica multiple nucleopolyhedrovirus (acmnpv), commonly used for large-scale protein p ... | 2009 | 19111552 |
| 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 ... | 2008 | 19149181 |
| acmnpv ac16 (da26, bv/odv-e26) regulates the levels of ie0 and ie1 and binds to both proteins via a domain located within the acidic transcriptional activation domain. | ie0 and ie1 are the primary viral regulatory proteins of autographa californica multiple nucleopolyhedrovirus (acmnpv) involved in the transactivation of early genes, stimulation of late gene expression, and viral dna replication. the protein interactions required for ie0 or ie1 to achieve these varied roles are not well defined, so to identify proteins that interact with ie0 and ie1, tandem affinity purification (tap) and lc-ms/ms was used. analysis of purified proteins identified ac16 (da26, b ... | 2009 | 19150105 |
| acmnpv exon0 (ac141) which is required for the efficient egress of budded virus nucleocapsids interacts with beta-tubulin. | the autographa californica multiple nucleopolyhedrovirus (acmnpv) encoded protein, exon0 (ac141), is required for the efficient transport of nucleocapsids out of the nucleus for the production of budded virus (bv). to further elucidate the molecular mechanisms by which exon0 regulates bv production, exon0 was tagged at the n-terminus with 3x flag-6x his. protein complexes were isolated by tandem affinity purification and potential exon0 specific interacting protein partners were gel purified and ... | 2009 | 19155039 |
| caspase inhibitor p35 is required for the production of robust baculovirus virions in trichoplusia ni tn-368 cells. | apoptosis can protect lepidopteran insects against baculovirus infection by limiting viral replication. baculoviruses counter this response by expressing anti-apoptotic proteins such as the caspase inhibitor p35, which is expressed by several baculoviruses including autographa californica mutiple nucleopolyhedrovirus (acmnpv). mutants of acmnpv that lack the p35 gene induce apoptosis in spodoptera frugiperda cells, and replication of these mutants is severely curtailed in s. frugiperda cell line ... | 2009 | 19218211 |
| microinjection of xenopus laevis oocytes. | microinjection of xenopus laevis oocytes followed by thin-sectioning electron microscopy (em) is an excellent system for studying nucleocytoplasmic transport. because of its large nucleus and high density of nuclear pore complexes (npcs), nuclear transport can be easily visualized in the xenopus oocyte. much insight into the mechanisms of nuclear import and export has been gained through use of this system (reviewed by panté, 2006). in addition, we have used microinjection of xenopus oocytes to ... | 2009 | 19238134 |
| cloning and expression of simian rotavirus spike protein (vp4) in insect cells by baculovirus expression system. | vp4 protein is as spikes on rotavirus outer capsid shell which is responsible for virus attachment to the host. vp4 induces production of neutralizing antibodies which could be used for serotyping of different isolates. | 2009 | 19252673 |
| biophysical characterization and unfolding of lef4 factor of rna polymerase from acnpv. | late expression factor 4 (lef4) is one of the four subunits of autographa californica nuclear polyhedrosis virus (acnpv) rna polymerase. lef4 was overexpressed in escherichia coli and recombinant protein was subjected to structural characterization. chemical induced unfolding of lef4 was investigated using intrinsic fluorescence, hydrophobic dye binding, fluorescence quenching, and circular dichroism (cd) techniques. the unfolding of lef4 was found to be a non-two state, biphasic transition. int ... | 2009 | 19274720 |
| aerosol infectivity of a baculovirus to trichoplusia ni larvae: an alternative larval inoculation strategy for recombinant protein production. | the baculovirus-insect expression system is a popular tool for recombinant protein production. the standard method for infecting insect larvae with recombinant baculovirus for protein production involves either feeding occlusion bodies or injecting budded virus into the cuticle. in this study, we showed that the recombinant autographa californica multiple nucleopolyhedrovirus (acmnpv) at titers >10(8) pfu/ml efficiently infected trichoplusia ni (t. ni) larvae through aerosol inoculation of budde ... | 2009 | 19334290 |
| two new cell lines originated from the embryos of clostera anachoreta (lepidoptera: notodontidae): characterization and susceptibility to baculoviruses. | two cell lines designated caf-clan i and caf-clan ii have been established from embryos of clostera anachoreta (lepidoptera: notodontidae) in tnm-fh medium containing 10% inactivated fetal bovine serum. caf-clan i consists of a mixture of three cell types: spherical cells, spindle-shaped cells, and giant cells. most of the cultured cells formed a suspension in the medium and were subcultured more than 60 passages. caf-clan ii mainly consists of spindle-shaped and spherical cells which attached t ... | 2009 | 19343458 |
| ac109 is required for the nucleocapsid assembly of autographa californica multiple nucleopolyhedrovirus. | orf109 (ac109) of autographa californica multiple nucleopolyhedrovirus (acmnpv) is a highly conserved gene in all sequenced baculovirus genomes, but its function is not known. this paper describes generation of an ac109 knockout virus (ac-ac109-ko-gp) and analyses of the influence of ac109 deletion on the virus replication in sf-9 cells so as to investigate the role of ac109 in the viral life cycle. results revealed that budded virus (bv) yields and occlusion body synthesis were completely block ... | 2009 | 19393701 |
| deletion of the acmnpv core gene ac109 results in budded virions that are non-infectious. | autographa californica multiple nucleopolyhedrovirus (acmnpv) ac109 is a core gene and its function in the virus life cycle is unknown. to determine its role in the baculovirus life cycle, we used the acmnpv bacmid system to generate an ac109 deletion virus (vac(109ko)). fluorescence and light microscopy showed that transfection of vac(109ko) results in a single-cell infection phenotype. viral dna replication is unaffected and the development of occlusion bodies in vac(109ko)-transfected cells e ... | 2009 | 19411088 |
| pathogenesis of autographa californica multiple nucleopolyhedrovirus in fifth-instar anticarsia gemmatalis larvae. | we have investigated infection and pathogenesis of autographa californica multiple nucleopolyhedrovirus (acmnpv) in anticarsia gemmatalis (velvetbean caterpillar) larvae using a lacz recombinant virus (acmnpv-hsp70/lacz) to track the temporal progression of infection in the midgut intestine and haemocoel. a. gemmatalis was highly resistant to fatal infection by occlusion bodies (obs; ld(50)>5.5 x 10(5) ob) and budded virus (bv; ld(50)>3 x 10(5) bv) administered via oral and systemic routes, resp ... | 2009 | 19423548 |
| baculovirus interactions in vitro and in vivo. | baculoviruses are promising viral insecticides and are safe for the environment. interaction of baculoviruses in vitro and in vivo is a basic molecular and ecological question that has practical applications in agriculture. cellular secretion is also a fundamental property in cell-cell communication. here, we review recent investigations on how baculoviruses interact with insect cells and insect hosts. we focus particularly on a new interaction mechanism in which a secretion from cells infected ... | 2009 | 19426856 |
| baculovirus induces type i interferon production through toll-like receptor-dependent and -independent pathways in a cell-type-specific manner. | autographa californica nuclear polyhedrosis virus (acnpv) is a double-stranded-dna virus that is pathogenic to insects. acnpv was shown to induce an innate immune response in mammalian immune cells and to confer protection of mice from lethal viral infection. in this study, we have shown that production of type i interferon (ifn) by acnpv in murine plasmacytoid dendritic cells (pdcs) and non-pdcs, such as peritoneal macrophages and splenic cd11c+ dcs, was mediated by toll-like receptor (tlr)-dep ... | 2009 | 19474102 |
| chicken hs4 insulator significantly improves baculovirus-mediated foreign gene expression in insect cells by modifying the structure of neighbouring chromatin in virus minichromosome. | the baculovirus autographa californica multiple nucleopolyhedrovirus (acmnpv) is widely used as a eukaryotic expression vector for protein production. in the current study, chicken beta-globin 5'-hs4 insulator (hs4) was placed downstream of the polyhedrin promoter-directed foreign gene expression cassette in acmnpv, and found to markedly increase the expression of target gene. when enhanced green fluorescence protein gene (egfp) was used as the reporter gene, cells infected by the recombinant vi ... | 2009 | 19539676 |
| bacterial, but not baculoviral infections stimulate hemolin expression in noctuid moths. | lepidopteran larvae are regularly infected by baculoviruses during feeding on infected plants. the differences in sensitivity to these infections can be substantial, even among closely related species. for example, the noctuids cotton bollworm (helicoverpa zea) and tobacco budworm (heliothis virescens), have a 1000-fold difference in sensitivity to autographa californica multiple nucleopolyhedrovirus (acmnpv) infection. recent data were interpreted to indicate that the lepidopteran immunoglobuli ... | 2009 | 19540262 |
| characterization of short-lived intermediates produced during replication of baculovirus dna. | in this report the short-lived dna replication intermediates produced in both uninfected and autographa californica multiple nucleopolyhedrovirus (acmnpv) infected spodoptera frugiperda cells were characterized. the methods used included pulse-labeling of dna in permiabilized cells, treatment of nascent dna with mung bean nuclease, and electrophoresis in neutral and alkaline agarose gels. in contrast to uninfected cells that produced a population of small dna fragments of about 200bp, a populati ... | 2009 | 19560496 |
| the role of the pi3k-akt signal transduction pathway in autographa californica multiple nucleopolyhedrovirus infection of spodoptera frugiperda cells. | many viruses activate the phosphatidylinositol 3-kinase (pi3k)-akt signaling pathway, thereby modulating diverse downstream signaling pathways associated with antiapoptosis, proliferation, cell cycling, protein synthesis and glucose metabolism, in order to augment their replication. to date, the role of the pi3k-akt pathway in baculovirus replication has not been defined. in the present study, we demonstrate that infection of sf9 cells with autographa californica multiple nucleopolyhedrovirus (a ... | 2009 | 19573890 |
| enhancing the multiplication of nucleopolyhedrovirus in vitro by manipulation of the ph. | insect nucleopolyhedroviruses (npvs) are studied widely as agents for biological control, as expression vectors for the production of heterologous proteins, and as transduction vectors for gene therapy applications. most of these applications rely on the existence of cell lines that allow in vitro multiplication of the virus. the influence of ph in the medium culture on the multiplication of semnpv, hearsnpv and acmnpv in different cell culture lines was investigated. the study showed a strong i ... | 2009 | 19576934 |
| bombyx mori nucleopolyhedrovirus orf34 is required for efficient transcription of late and very late genes. | homologs of autographa californica nucleopolyhedrovirus orf43 (ac43) are found in all group i and most group ii npvs, but their functions remain unknown. in bombyx mori npv (bmnpv)-infected bmn cells, bm34, a bmnpv homolog of ac43, is expressed as a late gene and its product is localized in the nucleus. to examine the role of bm34 during bmnpv infection, we constructed a bm34 deletion mutant (bmorf34d) and characterized its infectivity in bmn cells and b. mori larvae. bmorf34d produced far fewer ... | 2009 | 19651423 |
| genetic modification of a baculovirus vector for increased expression in insect cells. | generating large amounts of recombinant protein in transgenic animals is often challenging and has a number of drawbacks compared to cell culture systems. the baculovirus expression vector system (bevs) uses virus-infected insect cells to produce recombinant proteins to high levels, and these are usually processed in a similar way to the native protein. interestingly, since the development of the bevs, the virus most often used (autographa californica multi-nucleopolyhedovirus; acmnpv) has been ... | 2010 | 19655260 |
| the pre-transmembrane domain of the autographa californica multicapsid nucleopolyhedrovirus gp64 protein is critical for membrane fusion and virus infectivity. | the envelope glycoprotein, gp64, of the baculovirus autographa californica multicapsid nucleopolyhedrovirus (acmnpv) is a class iii viral fusion protein that mediates ph-triggered membrane fusion during virus entry. viral fusion glycoproteins from many viruses contain a short region in the ectodomain and near the transmembrane domain, referred to as the pre-transmembrane (ptm) domain. in some cases, the ptm domain is rich in aromatic amino acids and plays an important role in membrane fusion. al ... | 2009 | 19692475 |
| baculovirus dna replication-specific expression factors trigger apoptosis and shutoff of host protein synthesis during infection. | apoptosis is an important antivirus defense. to define the poorly understood pathways by which invertebrates respond to viruses by inducing apoptosis, we have identified replication events that trigger apoptosis in baculovirus-infected cells. we used rna silencing to ablate factors required for multiplication of autographa californica multicapsid nucleopolyhedrovirus (acmnpv). transfection with double-stranded rna (dsrna) complementary to the acmnpv late expression factors (lefs) that are design ... | 2009 | 19706708 |
| multigene expression of protein complexes by iterative modification of genomic bacmid dna. | many cellular multi-protein complexes are naturally present in cells at low abundance. baculovirus expression offers one approach to produce milligram quantities of correctly folded and processed eukaryotic protein complexes. however, current strategies suffer from the need to produce large transfer vectors, and the use of repeated promoter sequences in baculovirus, which itself produces proteins that promote homologous recombination. one possible solution to these problems is to construct bacul ... | 2009 | 19725957 |
| functional analysis of the autographa californica nucleopolyhedrovirus iap1 and iap2. | the autographa californica nucleopolyhedrovirus (acmnpv) contains three apoptosis suppressor genes: p35, iap1 and iap2. acmnpv p35 functions as a pancaspase inhibitor, but the function of iap1 and iap2 has not been entirely resolved. in this paper, we analyze the function of iap1 and iap2 in detail. acmnpv with p35-deletion inhibited the apoptosis of bti-tn-5b1-4 (tn-hi5) cells induced by a helicoverpa armigera single nucleocapsid npv (hearnpv) infection and rescued the replication of hearnpv an ... | 2009 | 19727595 |
| a point mutation in the polyhedrin gene of a baculovirus, autographa californica mnpv, prevents crystallization of occlusion bodies. | the polyhedrin gene region of the autographa californica nuclear polyhedrosis virus(acmnpv) morphology mutant m29 has been characterized by genetic and physical techniques. recombination analysis of mutants m29 and acm5polyl demonstrated that wild-type polyhedrin recombinants could be obtained and that the dna restriction patterns of the recombinant viruses were identical to wild-type acmnpv dna. marker rescue experiments using the wild-type acmnpv ecori i fragment indicated that the morphology ... | 1987 | 19731454 |
| autographa californica multiple nucleopolyhedrovirus core gene ac96 encodes a per os infectivity factor (pif-4). | autographa californica multiple nucleopolyhedrovirus (acmnpv) ac96 is a core gene, but its role in virus replication is still unknown. to determine its role in the baculovirus life cycle, we used the acmnpv bacmid system to generate an ac96-null virus (vac(96)(null)). our analyses showed that the absence of ac96 does not affect budded virus (bv) production or viral dna replication in infected sf9 cells. western blotting and confocal immunofluorescence analysis showed that ac96 is expressed in bo ... | 2009 | 19759145 |
| virion-associated viral fibroblast growth factor stimulates cell motility. | the autographa californica m nucleopolyhedrovirus (acmnpv) viral fibroblast growth factor (vfgf) has functional parallels to cellular fgfs. deletion of the acmnpv vfgf has no obvious phenotype in cell culture but delays the time of insect death. here, we determined vfgf production during virus infection. vfgf was detected at 24 hours post infection and through the remainder of the infection cycle. since vfgf is thought to be a secreted membrane-binding protein and virions acquire an envelope der ... | 2009 | 19800090 |
| cloning of circular dnas from microorganisms using a novel plasmid capture system. | plasmid capture system (pcs) facilitates cloning and manipulation of circular double-stranded dna. we recently developed an improved pcs (pcs-lz) to clone relatively large dna molecules of 30-150 kb. the pcs-lz donor consists of a mini-f replicon and a kanamycin resistance marker between tn7 left and tn7 right ends. both the replicon and marker gene of the pcs-lz donor are transferred into target plasmid dnas by in vitro transposition, followed by replication in e. coli. colonies are tested for ... | 2010 | 19838822 |
| recombinant proteoliposomes prepared using baculovirus expression systems. | proteoliposomes are useful for investigating the functions and properties of membrane proteins. we have established a novel method to prepare proteoliposomes using a baculovirus (autographa californica nuclear polyhedrosis virus; acnpv) gene expression system producing the recombinant membrane proteins to be reconstituted. this method consists of two key steps for production of recombinant proteoliposomes: (1) the cdna of the recombinant membrane protein on a baculovirus vector is transfected in ... | 2009 | 19913163 |
| how baculovirus polyhedra fit square pegs into round holes to robustly package viruses. | natural protein crystals (polyhedra) armour certain viruses, allowing them to survive for years under hostile conditions. we have determined the structure of polyhedra of the baculovirus autographa californica multiple nucleopolyhedrovirus (acmnpv), revealing a highly symmetrical covalently cross-braced robust lattice, the subunits of which possess a flexible adaptor enabling this supra-molecular assembly to specifically entrap massive baculoviruses. inter-subunit chemical switches modulate the ... | 2010 | 19959989 |
| expression from baculovirus and serological reactivity of the nucleocapsid protein of dolphin morbillivirus. | the nucleocapsid (n) protein of dolphin morbillivirus (dmv) was expressed from a baculovirus (autographa californica nuclear polyhedrosis virus) vector and shown by sds-page and western blot analysis to be about 57 kda. transmission electron microscopy revealed fully assembled nucleocapsid-like particles (nlps) exhibiting the typical helical herringbone morphology. these nlps were approximately 20-22 nm in diameter and varied in length from 50 to 100 nm. purified dmv-n protein was used as antige ... | 2010 | 20005643 |
| autographa californica multiple nucleopolyhedrovirus odv-e56 envelope protein is required for oral infectivity and can be substituted functionally by rachiplusia ou multiple nucleopolyhedrovirus odv-e56. | the autographa californica multiple nucleopolyhedrovirus (acmnpv) odv-e56 gene encodes an occlusion-derived virus (odv)-specific envelope protein, odv-e56. in a previous analysis, the odv-e56 gene was found to be under positive selection pressure, suggesting that it may be a determinant of virus host range. to assess the role of odv-e56 in oral infectivity and host range, we constructed recombinant acmnpv clones (ac69gfp-e56lacz and aciegfp-e56lacz) in which odv-e56 protein synthesis was elimina ... | 2010 | 20032203 |
| prorenin processing enzyme (ppe) produced by baculovirus-infected sf-9 insect cells: ppe is the cysteine protease encoded in the acmnpv gene. | in infection cultures of spodoptera frugiperda (sf-9) insect cells with a recombinant baculovirus, vhpr, carrying human preprorenin cdna in the polyhedrin locus of autographa californica multiple nuclear polyhedrosis virus (acmnpv), the expressed inactive recombinant human (rh)-prorenin is reported to be proteolytically processed to yield active rh-renin in the very late phase of culture (takahashi et al., biosci. biotechnol. biochem., 71, 2610-2613 (2007)). to identify the enzyme that catalyzes ... | 2010 | 20139610 |
| recombinant baculovirus as a highly potent vector for gene therapy of human colorectal carcinoma: molecular cloning, expression, and in vitro characterization. | present therapeutic strategies for most cancers are restricted mainly to the primary tumors and are also not very effective in controlling metastatic states. alternatively, gene therapy can be a potential option for treating such cancers. currently mammalian viral-based cancer gene therapy is the most popular approach, but the efficacy has been shown to be quite low in clinical trials. in this study, for the first time, the insect cell-specific baculovirus autographa californica multiple nucleop ... | 2010 | 20143184 |
| autographa californica multiple nucleopolyhedrovirus lef-2 is a capsid protein required for amplification but not initiation of viral dna replication. | the late expression factor 2 gene (lef-2) of baculovirus autographa californica multiple nucleopolyhedrovirus (acmnpv) has been identified as one of the factors essential for origin-dependent dna replication in transient expression assays and has been shown to be involved in late/very late gene expression. to study the function of lef-2 in the life cycle of acmnpv, lef-2 knockout and repair bacmids were generated by homologous recombination in escherichia coli. growth curve analysis showed that ... | 2010 | 20219928 |
| autographa californica multicapsid nucleopolyhedrovirus efficiently infects sf9 cells and transduces mammalian cells via direct fusion with the plasma membrane at low ph. | the budded virus (bv) of the autographa californica multicapsid nucleopolyhedrovirus (acmnpv) infects insect cells and transduces mammalian cells mainly through the endocytosis pathway. however, this study revealed that the treatment of the virus bound to sf9 cells at low ph could efficiently rescue the infectivity of acmnpv in the presence of endocytosis pathway inhibitors. a colocalization assay of the major capsid protein vp39 with the early endosome marker eea1 showed that at low ph, acmnpv ... | 2010 | 20219938 |
| purification and characterization of a viral chitinase active against plant pathogens and herbivores from transgenic tobacco. | the autographa californica nucleopolyhedrovirus chitinase a (acmnpv chia) is a chitinolytic enzyme with fungicidal and insecticidal properties. its expression in transgenic plants enhances resistance against pests and fungal pathogens. we exploited tobacco for the production of a biologically active recombinant acmnpv chia (rchia), as such species is an alternative to traditional biological systems for large-scale enzyme production. the protein was purified from leaves using ammonium sulfate pre ... | 2010 | 20302895 |
| 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 ... | 2008 | 20334593 |
| identification of a domain of the baculovirus autographa californica multiple nucleopolyhedrovirus single-strand dna-binding protein lef-3 essential for viral dna replication. | autographa californica multiple nucleopolyhedrovirus (acmnpv) lef-3 is one of nine genes required for viral dna replication in transient assays. lef-3 is predicted to contain several domains related to its functions, including nuclear localization, single-strand dna binding, oligomerization, interaction with p143 helicase, and interaction with a viral alkaline nuclease. to investigate the essential nature of lef-3 and the roles it may play during baculovirus dna replication, a lef-3 null bacmid ... | 2010 | 20357098 |
| baculovirus infection influences host protein expression in two established insect cell lines. | we identified host proteins that changed in response to host cell susceptibility to baculovirus infection. we used three baculovirus-host cell systems utilizing two cell lines derived from pupal ovaries, hz-am1 (from helicoverpa zea) and hv-am1 (from heliothis virescens). hv-am1 cells are permissive to autographa californica multiple nucleopolyhedrovirus (acmnpv) and semi-permissive to h. zea single nucleopolyhedrovirus (hzsnpv). hz-am1 cells are non-permissive to acmnpv. we challenged each cell ... | 2010 | 20362582 |
| the uga-cie1 cell line from chrysodeixis includens exhibits characteristics of granulocytes and is permissive to infection by two viruses. | the soybean looper, chrysodeixis (pseudoplusia) includens (lepidoptera: noctuidae) is an economically important insect pest and a highly permissive host for the parasitoid microplitis demolitor and its associated polydnavirus m. demolitor bracovirus (mdbv). here we established a cell line from c. includens embryos designated uga-cie1 cells. cie1 cells morphologically resemble granulocytes, which are a subpopulation of c. includens hemocytes. antibody and rt-pcr analyses indicated that cie1 cells ... | 2010 | 20381615 |
| host innate immune responses induced by baculovirus in mammals. | the baculovirus autographa californica nuclear polyhedrosis virus has been widely used not only to achieve a high level of foreign gene expression in insect cells but also for efficient gene transduction into mammalian cells without any replication. in addition to the efficient gene delivery, baculovirus has been shown to induce host innate immune responses in various mammalian cells and in mice. the baculovirus has abundant cpg motifs in the viral genome and is capable of inducing pro-inflammat ... | 2010 | 20394574 |
| baculovirus for eukaryotic protein display. | the baculovirus autographa californica nuclear polyhedrosis virus is an attractive candidate for eukaryotic virus display. a variety of strategies exists to incorporate and present target proteins on the surface of infected insect cells as well as on budded virions. native baculovirus proteins such as the major envelope protein and the capsid protein, but also foreign scaffolds such as the vesicular stomatitis virus g-1 protein or the influenza a virus hemagglutinin serve as fusion partners for ... | 2010 | 20433416 |
| functional analysis of the inhibitor of apoptosis genes in antheraea pernyi nucleopolyhedrovirus. | the inhibitor of apoptosis proteins (iap) plays an important role in cell apoptosis. we cloned two novel iap family members, ap-iap1 and ap-iap2, from antheraea pernyi nucleopolyhedrovirus (apnpv) genome. ap-iap1 contains two baculoviral iap repeat (bir) domains followed by a ring domain, but ap-iap2 has only one bir domain and ring. the result of transient expression in spodoptera frugiperda (sf21) showed that ap-iap1 blocked cell apoptosis induced by actinomycin d treatment and also rescued th ... | 2010 | 20437152 |
| proteomics of the autographa californica nucleopolyhedrovirus budded virions. | baculoviruses produce two progeny phenotypes during their replication cycles. the occlusion-derived virus (odv) is responsible for initiating primary infection in the larval midgut, and the budded virus (bv) phenotype is responsible for the secondary infection. the proteomics of several baculovirus odvs have been revealed, but so far, no extensive analysis of bv-associated proteins has been conducted. in this study, the protein composition of the bv of autographa californica nucleopolyhedrovirus ... | 2010 | 20444894 |
| a viral chitinase enhances oral activity of tmof. | in this study we investigate the combined effect on heliothis virescens (lepidoptera, noctuidae) larvae of aedes aegypti-trypsin modulating oostatic factor (aea-tmof), a peptide that inhibits trypsin synthesis by the gut, impairing insect digestive function, and autographa californica nucleopolyhedrovirus chitinase a (acmnpv chia), an enzyme that is able to alter the permeability of the peritrophic membrane (pm). aea-tmof and acmnpv chia were provided to the larvae by administering transgenic to ... | 2010 | 20457253 |
| establishment of a cell line from chrysodeixis chalcites permissive for chrysodeixis chalcites and trichoplusia ni nucleopolyhedrovirus. | a new cell line was established from the embryos of the insect chrysodeixis chalcites (lepidoptera, noctuidae, plusiinae). the cell line contains several morphologically different cell types and was distinguished from three other lepidopteran cell lines propagated in the laboratory by dna amplification fingerprinting. the cultured cells, which we officially named wu-cce-1 cells, were permissive for infection by c. chalcites nucleopolyhedrovirus (chchnpv) and large numbers of occlusion bodies wer ... | 2010 | 20471390 |
| autographa californica multiple nucleopolyhedrovirus ac76 is involved in intranuclear microvesicle formation. | in this study, we characterized autographa californica multiple nucleopolyhedrovirus (acmnpv) orf76 (ac76), which is a highly conserved gene of unknown function in lepidopteran baculoviruses. transcriptional analysis of ac76 revealed that transcription of multiple overlapping multicistronic transcripts initiates from a canonical taag late-transcription start motif but terminates at different 3' ends at 24 h postinfection in acmnpv-infected sf9 cells. to investigate the role of ac76 in the baculo ... | 2010 | 20484514 |
| the putative pocket protein binding site of autographa californica nucleopolyhedrovirus bv/odv-c42 is required for virus-induced nuclear actin polymerization. | nuclear filamentous actin (f-actin) is essential for nucleocapsid morphogenesis of lepidopteran nucleopolyhedroviruses. previously, we had demonstrated that autographa californica multiple nucleopolyhedrovirus (acmnpv) bv/odv-c42 (c42) is involved in nuclear actin polymerization by recruiting p78/83, an acmnpv orf9-encoded n-wasp homology protein that is capable of activating an actin-related-protein 2/3 (arp2/3) complex to initiate actin polymerization, to the nucleus. to further investigate th ... | 2010 | 20484515 |
| confocal microscopic observation of fusion between baculovirus budded virus envelopes and single giant unilamellar vesicles. | we assayed fusion events between giant unilamellar vesicles (guvs) and budded viruses (bvs) of baculovirus (autographa californica nucleopolyhedrovirus), the envelopes of which have been labeled with the fluorescent dye alexa fluor 488. this involves observing the intensity of fluorescence emitted from the lipid bilayer of single guvs after fusion using laser scanning microscopy. using this assay system, we found that fusion between single guvs and bv envelopes was significantly enhanced at arou ... | 2010 | 20493165 |
| characterization of a bombyx mori nucleopolyhedrovirus mutant lacking both fp25k and p35. | recent studies have shown that dual mutations in fp25k and p35 of autographa californica nucleopolyhedrovirus (acmnpv) result in a typical apoptotic infection on trichoplusia ni cells, suggesting the involvement of fp25k on npv-induced apoptosis. to examine the effect of fp25k deletion on bombyx mori npv (bmnpv)-induced apoptosis, we generated a bmnpv mutant, fp-p35d, in which both fp25k and p35 genes are deleted from the genome, and compared its phenotype with wild-type (t3), fp25k-deleted (fp- ... | 2010 | 20508980 |
| specificity of baculovirus p6.9 basic dna-binding proteins and critical role of the c terminus in virion formation. | the majority of double-stranded dna (dsdna) viruses infecting eukaryotic organisms use host- or virus-expressed histones or protamine-like proteins to condense their genomes. in contrast, members of the baculoviridae family use a protamine-like protein named p6.9. the dephosphorylated form of p6.9 binds to dna in a non-sequence-specific manner. by using a p6.9-null mutant of autographa californica multiple nucleopolyhedrovirus (acmnpv), we demonstrate that p6.9 is not required for viral dna repl ... | 2010 | 20519380 |
| [expression of baculovirus acmnpv p48 gene in insect cell]. | p48 (ac103) gene is highly conserved in baculovirus, implying that p48 might play a fundamental role in the life cycle of baculovirus. we studied the expression of p48 gene of autographa californica multiple nucleopolyhedrovirus (acmnpv)--the type baculovirus species. | 2010 | 20560348 |
| standardization of an elisa test using a recombinant nucleoprotein from the junin virus as the antigen and serological screening for arenavirus among the population of nova xavantina, state of mato grosso. | arenavirus hemorrhagic fever is a severe emerging disease. | 2010 | 20563486 |
| effect of ac68 knockout and lef3 leading sequence disruption on viral propagation. | orf68 (ac68) of autographa californica multiple nucleopolyhedrovirus (acmnpv) is identified to be an early gene, but its transcription start site remains unknown. the coding sequence of ac68 overlaps 280-bp leading sequence and 159-bp coding sequence of lef3 (ac67). in this study, the transcription start site of ac68 was determined by 5' race analysis to be 18 nucleotides upstream from the start codon. in order to investigate the effect of ac68 deletion on virus propagation, we generated a bacmi ... | 2011 | 20567974 |
| baculovirus gp64 disulfide bonds: the intermolecular disulfide bond of autographa californica multicapsid nucleopolyhedrovirus gp64 is not essential for membrane fusion and virion budding. | the gp64 envelope glycoprotein of the autographa californica nucleopolyhedrovirus (acmnpv) is a class iii viral membrane fusion protein that is triggered by low ph during entry. unlike most other viral fusion protein trimers, the monomers of gp64 are covalently linked to each other within the trimer by a single intermolecular disulfide bond (cys24 cys372). single or paired alanine substitutions for cys24 and cys372 resulted in lower-efficiency transport of gp64 to the cell surface. surprisingly, ... | 2010 | 20573818 |
| baculovirus treatment fully protects mice against a lethal challenge of fmdv. | foot-and-mouth disease virus (fmdv) causes a highly contagious and economically devastating disease that affects cattle, swine, goat and sheep among others. fmdv is able to overcome the initial host innate immune response by inhibiting the induction of antiviral molecules at both the transcriptional and the translational levels. it has been demonstrated that fmdv a/arg/2001 causes the death of adult c57bl/6 mice within 72h. we evaluated the capacity of autographa californica nuclear polyhedrosis ... | 2010 | 20580746 |
| baculovirus per os infectivity factors form a complex on the surface of occlusion-derived virus. | five highly conserved per os infectivity factors, pif1, pif2, pif3, pif4, and p74, have been reported to be essential for oral infectivity of baculovirus occlusion-derived virus (odv) in insect larvae. three of these proteins, p74, pif1, and pif2, were thought to function in virus binding to insect midgut cells. in this paper evidence is provided that pif1, pif2, and pif3 form a stable complex on the surface of odv particles of the baculovirus autographa californica multinucleocapsid nucleopolyh ... | 2010 | 20610731 |
| recombinant peroxidase production in species of lepidoptera frequently found in argentina. | horseradish peroxidase isozyme c (hrpc) is an important commercial biocatalyst. in this study, a screening of different lepidopteran species frequently found in argentina to produce this protein was carried out. two recombinant viruses were constructed: acmnpv hrpc polyhedrin-minus (occ-), an intrahemocelical infective virus; and acmnpv hrpc polyhedrin-plus (occ+), to achieve an oral infective baculovirus. each lepidopteran species was infected either with acmnpv hrpc occ- or acmnpv hrpc occ+ an ... | 2010 | 20615485 |
| identification and functional analysis of the putative anti-apoptotic gene iap4 of spodoptera litura nucleopolyhedrovirus. | baculoviruses possess two types of anti-apoptotic genes, p35 and inhibitor of apoptosis (iap). in this study, we cloned a novel iap family member, splt-iap4, from spodoptera litura nucleopolyhedrovirus (spltnpv). scanprosite analysis of splt-iap4 amino acid sequence show that it contain a baculovirus iap repeat (bir) motif at the n-terminal portion from 18 to 61 amino acid and a zinc finger ring motif near its c-terminus. baculovirus iaps could be divided into five types from iap1 to iap5 accord ... | 2010 | 20625923 |
| a single amino acid substitution modulates low-ph-triggered membrane fusion of gp64 protein in autographa californica and bombyx mori nucleopolyhedroviruses. | we have previously shown that budded viruses of bombyx mori nucleopolyhedrovirus (bmnpv) enter the cell cytoplasm but do not migrate into the nuclei of non-permissive sf9 cells that support a high titer of autographa californica multicapsid nucleopolyhedrovirus (acmnpv) multiplication. here we show, using the syncytium formation assay, that low-ph-triggered membrane fusion of bmnpv gp64 protein (bm-gp64) is significantly lower than that of acmnpv gp64 protein (ac-gp64). mutational analyses of gp ... | 2010 | 20627345 |
| deletion of acmnpv ac16 and ac17 results in delayed viral gene expression in budded virus infected cells but not transfected cells. | this study investigated the combined function of the autographa californica multiple nucleopolyhedrovirus overlapping genes ac16 (bv/odv-e26, da26) and ac17. ac17 is a late gene and the promoter is within the ac16 open reading frame. a double ac16-ac17 knockout virus was generated to assess the function of each gene independently or together. loss of ac17 did not affect viral dna synthesis but budded virus (bv) production was reduced. deletion of both ac16-ac17 resulted in reduced viral dna synt ... | 2010 | 20627351 |
| functional analysis of spodoptera litura nucleopolyhedrovirus p49 gene during autographa californica nucleopolyhedrovirus infection of spli-221 cells. | the spodoptera litura nucleopolyhedrovirus (spltnpv) antiapoptotic gene splt-p49 is able to rescue replication of a p35-null autographa californica nucleopolyhedrovirus (acmnpv) in acmnpv-permissive sf9 cells. in this study, an acmnpv p35 knockout bacmid was generated through et homologous recombination in escherichia coli and designated as vac(p35-ko). the splt-p49 gene was transposed into the polyhedrin locus of vac(p35-ko) to investigate if splt-p49 has any antiapoptotic activity in the conte ... | 2010 | 20711805 |
| partial functional rescue of helicoverpa armigera single nucleocapsid nucleopolyhedrovirus infectivity by replacement of f protein with gp64 from autographa californica multicapsid nucleopolyhedrovirus. | two distinct envelope fusion proteins (efps) (gp64 and f) have been identified in members of the baculoviridae family of viruses. f proteins are found in group ii nucleopolyhedroviruses (npvs) of alphabaculoviruses and in beta- and deltabaculoviruses, while gp64 occurs only in group i npvs of alphabaculoviruses. it was proposed that an ancestral baculovirus acquired the gp64 gene that conferred a selective advantage and allowed it to evolve into group i npvs. the f protein is a functional analog ... | 2010 | 20739531 |
| inorganic binding peptide-mediated immobilization based on baculovirus surface display system. | the biomolecule-mediated assembly of novel composites has been the subject of numerous investigations during the last years, providing new insights into material science and engineering. via molecular biology technology, we were able to introduce the genetically engineered polypeptide for inorganics (gepi) as a molecular binder into biomolecules such as phage viruses, to assemble hybrid functional nanoarchitectures. in the present work, we introduced a novel nanocomposite comprising the autograp ... | 2010 | 20806244 |
| removal of transposon target sites from the autographa californica multiple nucleopolyhedrovirus fp25k gene delays, but does not prevent, accumulation of the few polyhedra phenotype. | low-cost, large-scale production of the baculovirus autographa californica multiple nucleopolyhedrovirus (acmnpv) using continuous insect cell culture is seriously hindered by the accumulation of acmnpv mutants. specifically, few-polyhedra (fp) mutants, with a reduced yield of occluded virus (polyhedra) and decreased infectivity, usually accumulate upon passaging in cell culture. fp mutations result from transposon insertions in the baculovirus fp25k gene, leading to significantly reduced levels ... | 2010 | 20810745 |
| comparative transient expression assay analysis of hycu-hr6- and ie1-dependent regulation of baculovirus gp64 early promoters in three insect cell lines. | we previously demonstrated that the hyphantria cunea multicapsid nucleopolyhedrovirus (hycumnpv) gp64 gene (hycu-gp64) is uniquely localized on the viral genome with a large homologous region of 1582bp, hycu-hr6, immediately upstream of the hycu-gp64 gene. in the present study, we compared the regulation of gp64 early promoters from hycumnpv, autographa californica multicapsid npv (acmnpv) and bombyx mori npv (bmnpv) by cis-acting hycu-hr6 and trans-acting ie1s in three cell lines (spodoptera fr ... | 2011 | 20837073 |
| effect of starvation upon baculovirus replication in larval bombyx mori and heliothis virescens. | the progression of baculovirus (bmnpv, bmcyspd, acmnpv or acaait) infection in larval bombyx mori and heliothis virescens (1st, 3rd or 5th instar) was investigated following various starvation regimes. when the larvae were starved for 12 or 24h immediately following inoculation, the median lethal time to death (lt(50)) was delayed by 9.5-19.2h in comparison to non-starved controls. this corresponded to a delay of 10-23% depending upon the larval stage and virus that was used for inoculation. whe ... | 2011 | 20850449 |
| autographa californica multiple nucleopolyhedrovirus core gene ac92 (p33) is required for efficient budded virus production. | autographa californica multiple nucleopolyhedrovirus (acmnpv) ac92 is a core gene encoding a protein associated with occlusion derived virus (odv), binds human p53 and also has flavin adenine dinucleotide linked sulfhydryl oxidase activity but its role in the virus life cycle is not known. to determine ac92 function a deletion virus (vac(92ko)) was generated and transfected sf9 cells revealed that vac(92ko) infection was restricted primarily to single cells and budded virus (bv) titer was reduce ... | 2010 | 20965540 |
| autographa californica multiple nucleopolyhedrovirus odv-e56 is a per os infectivity factor, but is not essential for binding and fusion of occlusion-derived virus to the host midgut. | the autographa californica multiple nucleopolyhedrovirus (acmnpv) occlusion-derived virus (odv) envelope protein odv-e56 is essential for oral infection of larvae of heliothis virescens. bioassays with recombinant clones of acmnpv lacking a functional odv-e56 gene showed that odv-e56 was required for infectivity of both polyhedra and to a lesser extent, purified odv. however, binding and fusion assays showed that odv lacking odv-e56 bound and fused to midgut cells at levels similar to odv of wil ... | 2011 | 20970820 |
| a silencing suppressor protein (nss) of a tospovirus enhances baculovirus replication in permissive and semipermissive insect cell lines. | the nonstructural protein (nss) of the tomato spotted wilt virus (tswv) has been identified as an rnai suppressor in plant cells. a recombinant autographa californica multiple nucleopolyhedrovirus (acmnpv) designated vacnss, containing the nss gene under the control of the viral polyhedrin (polh) gene promoter, was constructed and the effects of nss in permissive, semipermissive and nonpermissive insect cells to vacnss infection were evaluated. vacnss produced more budded virus when compared to ... | 2010 | 20971139 |
| characterization of an autographa californica multiple nucleopolyhedrovirus mutant lacking the ac39(p43) gene. | open reading frame 39 [orf39(p43)] of autographa californica multicapsid nucleopolyhedrovirus (acmnpv) is present in 10 isolates of the alphabaculovirus genus. it is highly conserved in sequence and genomic location in the group i but much less conserved in the group ii viruses. to investigate the potential role of p43 in acmnpv infection, we constructed and characterized a p43 knockout mutant. the results showed that the p43 region was expressed as rna from 3h post infection to at least 24h pos ... | 2010 | 20974197 |
| [expression and characterization of rabies virus nucleoprotein in baculovirus]. | to construct a recombinant expression plasmid bacmid-n containing the n gene of rabies virus, the n gene of rv cvs-11 strain was cloned into the baculovirus shuttle vector (bacmid). recombinant baculovirus acmnpv-n (p1 viral stock) was obtained by transfecting the bacmid-n into the insect cell line of sf9. the expressed nucleoprotein was identified and analysised by elisa, fa, sds-page and western blot assays. the results showed that the np protein was expressed intracellularly and had a good an ... | 2010 | 21043133 |
| benzylideneacetone, an eicosanoid biosynthesis inhibitor enhances baculovirus pathogenicity in the diamondback moth, plutella xylostella. | benzylideneacetone (bza) is a monoterpenoid compound produced by an entomopathogenic bacterium, xenorhabdus nematophila. bza inhibits phospholipase a(2) to suppress biosynthesis of eicosanoids that mediate immune responses in insects. in response to per os infection of autographa californica multiple nucleopolyhedrosis virus (acmnpv), the diamondback moth, plutella xylostella, developed red spots on the midgut epithelium. the midgut exhibiting red spot formation suffered abnormal cell integrity, ... | 2010 | 21112333 |
| a peptide with similarity to baculovirus odv-e66 binds the gut epithelium of heliothis virescens and impedes infection with autographa californica multiple nucleopolyhedrovirus. | baculoviruses infect their lepidopteran hosts via the midgut epithelium through binding of occlusion-derived virus (odv) and fusion between the virus envelope and microvillar membranes. to identify genes and sequences that are involved in this process, a random phage display library was screened for peptides that bound to brush border membrane vesicles (bbmv) derived from the midgut epithelium of heliothis virescens. seventeen peptides that bound to bbmv were recovered. two of these, hv1 and hv2 ... | 2011 | 21228132 |
| comprehensive analysis of host gene expression in autographa californica nucleopolyhedrovirus-infected spodoptera frugiperda cells. | autographa californica multicapsid nucleopolyhedrovirus (acmnpv) is the best-studied baculovirus and most commonly used virus vector for baculovirus expression vector systems. the effect of acmnpv infection on host cells is incompletely understood. a microarray based on spodoptera frugiperda ests was used to investigate the impact of acmnpv on host gene expression in cultured s. frugiperda, sf21 cells. most host genes were down-regulated over the time course of infection, although a small number ... | 2011 | 21276998 |
| interaction of autographa californica multiple nucleopolyhedrovirus cathepsin protease progenitor (prov-cath) with insect baculovirus chitinase as a mechanism for prov-cath cellular retention. | the insect baculovirus chitinase (chia) and cathepsin protease (v-cath) enzymes cause terminal host insect liquefaction, enhancing the dissemination of progeny virions away from the host cadavers. regulated and delayed cellular release of these host tissue-degrading enzymes ensures that liquefaction starts only after optimal viral replication has occurred. baculoviral chia remains intracellular due to its c-terminal kdel endoplasmic reticulum (er) retention motif. however, the mechanism for cell ... | 2011 | 21289117 |
| identification of a cis-regulatory element that directs prothoracicotropic hormone gene expression in the silkworm bombyx mori. | in the silkworm bombyx mori and other insects, prothoracicotropic hormone (ptth) plays a central role in controlling molting and metamorphosis by stimulating the prothoracic glands to synthesize and release the molting hormone ecdysone. using an acnpv (autographa californica nucleopolyhedrovirus)-mediated transient gene transfer system, we identified a cis-regulatory element that participates in the decision to switch expression of ptth on or off in ptth-producing neurosecretory cells (ptpcs). t ... | 2011 | 21324358 |
| generation of recombinant baculovirus dna in e.coli using a baculovirus shuttle vector. | baculovirus vectors are now widely used to direct the expression of heterologous genes in cultured insect cells and insect larvae. in most cases, heterologous genes placed under transcriptional control of the polyhedrm promoter of the autographa californica nuclear polyhedrosis virus (acnpv) are abundantly expressed during the late stages of infection. the recombinant proteins are usually soluble and functionally similar to their authentic counterparts (l-7). in the following sections, recent ad ... | 1998 | 21390847 |
| establishment and characterization of an embryonic cell line from gampsocleis gratiosa (orthoptera: tettigoniidae). | the first continuous cell line from the embryo of gampsocleis gratiosa (orthoptera: tettigoniidae), designated as riri-gg1, was established. this cell line was serially subcultured in modified grace medium. the cells were grown adherent to a culture flask and had spindle-like and polygonal shapes. the chromosome number ranged from 26 to 79 at the 50th passage, and 68% of cells had a diploid chromosome number. the growth rate was determined at the 53rd passage, and the population doubling time wa ... | 2011 | 21424241 |
| genetic variation and virulence of nucleopolyhedroviruses isolated worldwide from the heliothine pests helicoverpa armigera, helicoverpa zea, and heliothis virescens. | to assess the diversity and relationships of baculoviruses found in insects of the heliothine pest complex, a pcr-based method was used to classify 90 samples of nucleopolyhedrovirus (npv; baculoviridae: alphabaculovirus) obtained worldwide from larvae of heliothis virescens, helicoverpa zea, and helicoverpa armigera. partial nucleotide sequencing and phylogenetic analysis of three highly conserved genes (lef-8, lef-9, and polh) indicated that 67 of these samples contained isolates of the h. zea ... | 2011 | 21439295 |
| autographa californica multiple nucleopolyhedrovirus odv-e66 is an essential gene required for oral infectivity. | autographa californica multiple nucleopolyhedrovirus (acmnpv) odv-e66 is a core gene and encodes an occlusion-derived virus (odv)-specific envelope protein, odv-e66. the n-terminal 23 amino acid of the envelope protein odv-e66 are sufficient to direct native and fusion proteins to induced membrane microvesicles and the viral envelope during infection with acmnpv. in this study, an odv-e66-knockout bacmid can not express n-terminal hydrophobic domains was constructed via homologous recombination ... | 2011 | 21440017 |
| baculovirus vp80 protein and the f-actin cytoskeleton interact and connect the viral replication factory with the nuclear periphery. | recently, we showed that the autographa californica multicapsid nucleopolyhedrovirus (acmnpv) vp80 protein is essential for the formation of both virion types, budded virus (bv) and occlusion-derived virus (odv). deletion of the vp80 gene did not affect assembly of nucleocapsids. however, these nucleocapsids were not able to migrate from the virogenic stroma to the nuclear periphery. in the current paper, we constructed a baculovirus recombinant with enhanced-green fluorescent protein (egfp)-tag ... | 2011 | 21450830 |
| baculoviruses mediate efficient gene expression in a wide range of vertebrate cells. | baculovirus expression vector system (bevs) is well known as a feasible and safe technology to produce recombinant (re-)proteins in a eukaryotic milieu of insect cells. however, its proven power in gene delivery and gene therapy is still poorly recognized. the basis of bevs lies in large enveloped dna viruses derived from insects, the prototype virus being autographa californica multiple nucleopolyhedrovirus (acmnpv). infection of insect cell culture with a virus encoding a desired transgene und ... | 2011 | 21590402 |
| the autographa californica multiple nucleopolyhedrovirus lef-5 gene is required for productive infection. | to examine the role of the acmnpv lef-5 gene in the context of the infection cycle, we generated an acmnpv lef-5 knockout virus (vac(lef5ko)) and a complementing cell line that supports viral replication. we examined acmnpv dna replication, early and late gene expression, and production of infectious viral progeny in the absence of lef-5. while early gene expression and dna replication were not reduced by the lef-5 knockout, expression of a late reporter was disrupted and representative late tra ... | 2011 | 21601232 |