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surface functionalized amorphous nanosilica and microsilica with nanopores as promising tools in biomedicine.cellular interactions with engineered nanoparticles (nps) are dependent on many properties, inherent to the nanoparticle (viz. size, shape, surface characteristics, degradation, agglomeration/dispersal, and charge, etc.). modification of the surface reactivity via surface functionalization of the nanoparticles to be targeted seems to be important. utilization of different surface functionalization methods of nanoparticles is an emerging field of basic and applied nanotechnology. it is well known ...200918791695
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
transgenic breeding of anti-bombyx mori l. nuclear polyhedrosis virus silkworm bombyx mori.silkworm strains resistant to bombyx mori l. nuclear polyhedrosis virus were obtained through transgenic experiments. piggybac transposon with an a3 promoter were randomly inserted into the silkworm, driving the enhanced green fluorescent protein (egfp) reporter gene into the silkworm genome. polymerase chain reaction results verified the insertion of the extraneous egfp gene, and fluorescence microscopy showed that the egfp was expressed in the midgut tissue. the morbidity ratio of the nuclear ...200818850052
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
biochemical analysis of hyphantria cunea npv attachment to spodoptera frugiperda 21 cells.binding characteristics of hyphantria cunea nuclear polyhedrosis virus (hcnpv) to spodoptera frugiperda 21 (sf21) cells was determined. the cells displayed an affinity of 0.9 x 10(10) m(-1) with about 8900 binding sites per cell. the biochemical nature of hcnpv-binding sites on the cell surface was also partially elucidated. there were 45 to 49% reductions in hcnpv binding following the pretreatment of cells with three proteases, suggesting the involvement of a cellular protein component in viru ...199919003136
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 ...200919066730
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.200919252673
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 ...200919274720
activity and persistence of steinernema carpocapsae and spodoptera exigua nuclear polyhedrosis virus against s. exigua larvae on soybean.experiments were conducted to assess the effects of the entomopathogenic nematode steinernema carpocapsae and spodoptera exigua multinucleocapsid nuclear polyhedrosis virus (semnpv), alone and in combinations, on mortality of the beet armyworm, s. exigua, larvae on soybean. in 1991 tests, field-grown soybean plants were treated with s. carpocapsae at 0.3 and 0.6 nematodes/cm(2) of leaflet, semnpv at 20 and 40 polyhedral inclusion bodies (pib)/cm(2), and all possible combinations. treated leaflet ...199619277347
an inducible system for highly efficient production of recombinant adeno-associated virus (raav) vectors in insect sf9 cells.production of clinical-grade gene therapy vectors for human trials remains a major hurdle in advancing cures for a number of otherwise incurable diseases. we describe a system based on a stably transformed insect cell lines harboring helper genes required for vector production. integrated genes remain silent until the cell is infected with a single baculovirus expression vector (bev). the induction of expression results from a combination of the amplification of integrated resident genes (up to ...200919279219
upregulation of proinflammatory cytokines and no production in bv-activated avian macrophage-like cell line (hd11) requires mapk and nf-kappab pathways.the budded virus (bv) of antheraea pernyi is a nuclear polyhedrosis virus (apnpv), that belongs to the baculovirus family. it is capable of stimulating innate immune response in avian macrophage-like cells (hd11). in this study, we used the expression of proinflammatory cytokines (ifn-gamma and il-12p40) and production of nitric oxide (no) as indicators to evaluate the involvement of the signaling cascades in the activation of hd11 cells by bv. western blot analysis revealed that erk1/2, p38 map ...200919303463
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 ...200919474102
[the effect of fodder on the susceptibility of gypsy moth (lymantria dispar l.) to nuclear polyhedrosis virus].gypsy moth (lymantria dispar l.) growing on different feeding substrates was shown to affect their susceptibility to nuclear polyhedrosis virus (npv). the insects feeding on birch leaves had the lowest sensitivity to npv than those on willow leaves, but the insects growing on pine needles showed the highest susceptibility. the sensitivity of the gypsy moths on willow leaves was higher than that of the gypsy moths on birch leaves and lower than that of those on pine needles. at the same time, it ...200919708555
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 ...198719731454
effects of insect viruses and pesticides on glutathione s-transferase activity and gene expression in bombyx mori.glutathione s-transferase (gst) is a gene family generally associated with detoxification and plays an important role in detoxifying exogenous compounds. the silkworm bombyx mori is an important economic animal for silk production. however, it is liable to infection by a number of viruses and chemical agents that can contaminate its food and growing environment. here we conducted a comparative study of strain- and tissue-specific expressions of gst in the silkworm under infections by bombyx mori ...200919736773
inhibition of influenza virus by baculovirus-mediated shrna.influenza viruses a and b cause widespread infections of the human respiratory tract; however, existing vaccines and drug therapy are of limited value for their treatment. here we show that bispecific short hairpin small-interfering rna constructs containing an eight-nucleotide intervening spacer, targeted against influenza virus a or influenza virus b, can inhibit the production of both types of virus in infected cell lines. this multiple vector showed remarkable ability to cope with both influ ...200919749373
baculovirus-mediated bispecific short-hairpin small-interfering rnas have remarkable ability to cope with both influenza viruses a and b.influenza viruses a and b cause widespread infections of the human respiratory tract; however, existing vaccines and drug therapy are of limited value for their treatment. here, we show that bispecific short-hairpin small-interfering rna constructs containing an 8-nucleotide intervening spacer, targeted against influenza virus a or influenza virus b, can inhibit the production of both types of virus in infected cell lines. this multiple vector showed remarkable ability to cope with both influenz ...200919899951
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 ...200919913163
nanoparticle-virus complex shows enhanced immunological effect against baculovirus.insects protect themselves from majority of infections by a non-specific innate immune system (present in both vertebrates and invertebrates). bombyx mori nuclear polyhedrosis virus (bmnpv), a baculovirus, causing the deadly grasserie disease is a scourge to silkworm industry and we report here the first success in combating this disease with the help of a nanosilica-virus complex. hydrophobic aluminium silicate nanoparticles were mixed with live bmnpv in vitro. this mixture was injected into on ...200919928266
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 ...201020005643
expression of ureb and hspa of helicobacter pylori in silkworm pupae and identification of its immunogenicity.for mass production of urease b subunit (ureb) and heat shock protein a subunit (hspa) of helicobacter pylori with bombyx mori nuclear polyhedrosis virus (bmnpv) baculovirus expression system (bes) and to determine whether they could be used as an oral vaccine against h. pylori, besides, to determine the time course of expressed recombinant protein and the optimum acquisition time directly through green fluorescence, hspa and enhanced green fluorescence protein (egfp) genes were cloned into vect ...201120127514
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 ...201020139610
phenotypic and genetic characteristic of nuclear polyhedrosis virus (npv) isolates from gypsy moth (lymantria dispar l.) larvae in natural populations of western siberia. 201020380159
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 ...201020394574
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 ...201020433416
[integration of influenza a virus nsp gene into baculovirus genome and its expression in insect cells].segment ns in all human influenza a viruses and in the part of avian and animal ones was found to contain an additional (beside ns1 and nep) long open reading frame (orf) enabling to code a polypeptide of 18-26 kd in different strains. this orf, in contrast to the ns1 and nep, locates in positive sense orientation in the negative polarity genomic ns rna segment and the encoded protein is referred nsp (negative strand protein). here, the nsp gene of human influenza a/wsn/33 (h1n1) virus was inser ...201020455463
[phenotypic and genetic characterization of nuclear polyhedrosis virus isolated from gypsy moth (lymantria dispar l.) larvae from the natural populations of western siberia].six nuclear polyhedrosis virus (npv) isolates have been isolated from dead larvae of gypsy moth in western siberia. heterogeneity of virulence and reproduction activity was revealed for the npv isolated by bioassay with lymantria dispar l. larvae. the findings may suggest phenotypic variation of the npv isolates. no correlation was found between virulence and reproductive activity with the only exception--the isolate karassuk with a high virulence and a high reproductive activity. nucleotide seq ...201020455470
involvement of tlr21 in baculovirus-induced interleukin-12 gene expression in avian macrophage-like cell line hd11.our previous work showed that baculovirus (antheraea pernyi nuclear polyhedrosis virus [apnpv]) induced a strong innate immunity and protected chicken from a lethal challenge of bronchitis virus. the purpose of present study was to determine the chicken toll-like receptors (tlrs) used by bv-infected immune cells to induce this immune response. we first investigated the expression of the toll-like receptor (tlr) genes in resting and bv-infected hd11 (chicken macrophage-like cell line) and dt40 (c ...201020471186
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 ...201020580746
protective effect of partially purified 35 kda protein from silk worm (bombyx mori) fecal matter against carbon tetrachloride induced hepatotoxicity and in vitro anti-viral properties.it has been found that many proteins from silkworm (bombyx mori l.) fecal matter have been active against human immunodeficiency virus, sendai virus, herpes simplex virus type-1, and nuclear polyhedrosis virus.201020738178
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 ...201020806244
high-level expression and improved folding of proteins by using the vp39 late promoter enhanced with homologous dna regions.some recombinant proteins expressed by baculovirus expression vector systems (bevs) aggregate because the bevs can produce large amounts of protein late during infection, when post-translational modification and protein quality control mechanisms are inactive. for expression during earlier stages than that driven by the polyhedrin (polh) very late promoter, transfer vectors were generated in which this promoter was replaced with a green fluorescent protein (gfp) gene controlled by a vp39 late pr ...201020821250
molecular cloning and characterization of twist gene in bombyx mori.the twist genes are an evolutionarily conserved group of regulatory basic helix-loop-helix (bhlh) transcription factors. in present study, the twist gene was firstly cloned from bombyx mori and was designated as bmtwist. sequence analysis showed that bmtwist cdna contains a 798 bp open reading frame, encoding a peptide of 266 amino acid residues. sequence alignment showed that bmtwist protein shared extensive homology with other invertebrate twist proteins in bhlh motif. rt-pcr and western blot ...201021076855
nanoparticle surface as activation site.the immense surface-to-volume (s/v) ratio in nanoparticles leads to large surface energy density. these high densities play the role of sites for activities that are not triggered in bulk materials. here we present some examples of such distinctive activities taking place at nanoparticle surfaces. our first example involves the morphological changes in silkworm (bombyx mori l.) nuclear polyhedrosis virus (bmnpv) brought about by lipophilic amorphous silica nanoparticles (lasn). microscopy studie ...201021137870
silkworm salivary glands are not susceptible to bombyx mori nuclear polyhedrosis virus.a nuclear polyhedrosis virus isolated from infected bombyx mori, bmnpv, was used to inoculate silkworms to determine salivary gland cell susceptibility. the salivary gland was removed from infected silkworms at different times post-inoculation and examined by light microscopy. the salivary gland cells did not exhibit any signs of bmnpv infection; however, fat body and tracheal cells, used as positive controls, showed characteristic cytopathological changes caused by bmnpv infection, which confir ...201121365549
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 ...199821390847
establishment, growth kinetics, and susceptibility to acmnpv of heat tolerant lepidopteran cell lines.lepidopteran heat-tolerant (ht) cell lines have been obtained with sf-9, sf-21 and several bombyx cells. they have a distinct karyotype, membrane lipid composition, morphology and growth kinetics from the parental cell lines. in this paper, we report the development of ht cell lines from other insect species and examination of their growth characteristics and virus susceptibility. adaptation of cell lines sf-9, bti-tn-5b1-4 (high5) and bti-tn-mg1 (mg1) to 33-¦c and 35-¦c was carried out by shift ...201121667340
[small rnas in bombyx mori nuclear polyhedrosis virus polyhedra form complexes with polypeptide p14 and polyhedrin].it was shown that two small rnas about 65 and 55 nucleotides long included in npv b. mori polyhedra form with polypeptides p29 and p14 specific rnp-complexes with molecular weights of 50 and 31 kda, respectively. both complexes form high-molecular weight complex with polyhedrin. origin and nature of p29 and p14 polypeptides are discussed.201021805866
secretory expression of all 16 subtypes of the hemagglutinin 1 protein of influenza a virus in insect cells.influenza viruses are known for their ability to change their antigenic structure and create new viral strains. hemagglutinin (ha), for which 16 subtypes have been identified, is a major antigenic determinant essential for the pathogenesis of influenza a viruses. to predict and monitor future epidemics, it is critical to produce various ha subtype antigens conveniently and rapidly. a simple, effective, and economic method to generate subunit ha1 of all 16 ha subtypes as recombinant proteins (rha ...201121827791
baculovirus capsid display potentiates ova cytotoxic and innate immune responses.baculoviruses (bv) are dna viruses that are pathogenic for insects. although bv infect a range of mammalian cell types, they do not replicate in these cells. indeed, the potential effects of these insect viruses on the immune responses of mammals are only just beginning to be studied. we show in this paper that a recombinant autographa californica multiple nuclear polyhedrosis virus carrying a fragment of ovalbumin (ova) on the vp39 capsid protein (bv-ova) has the capacity to act as an adjuvant ...201121918683
stable protein expression in mammalian cells using baculoviruses.the baculovirus autographa californica multiple nuclear polyhedrosis virus (acmnpv) has been widely used in biotechnology for protein expression in insect cells. baculoviruses use arthropods as their natural hosts and are unable to replicate in mammalian cells. however, acmnpv is able to enter many mammalian cell types and can be used for transgene expression if engineered to contain suitable expression cassettes. in this chapter, we describe the construction and application of a recombinant bac ...201221987248
modulation of innate immunity in chickens induced by in vivo administration of baculovirus.baculoviruses stimulate cytokine production in mammalian cells. they induce a strong innate immune response in animals and have adjuvant properties. the purpose of this work was to study the in vivo effect of baculovirus on chicken innate immune response. spf chickens were inoculated intravenously with autographa californica nuclear polyhedrosis virus (bv). three hours later, chickens were bled, euthanized and their spleen, duodenum and cecal tonsils were excised in order to take samples for rna ...201122142984
[the search of microrna genes encoded in antiparallel chains of autographa californica nuclear polyhedrosis virus genome regions, including most late and complementary genes].it was shown using bioinformatic approach by analysis of alternative transcriptes secondary structure, that a. californica nuclear polyhedrosis virus gene ph encoded two mature mirnas and three potential mirnas. gene orf1629 complementary to gene ph did not encode mirnas and pre-mirna-cs. gene p10 encodes mature and potential mirna. gene p74 located on complementary chain encodes three mature mirnas.201122145408
simulated acid rain reduces the susceptibility of the european pine sawfly (neodiprion sertifer) to its nuclear polyhedrosis virus.the study dealt with the effect of simulated acid rain (both h(2)so(4) and hno(3); acidities of ph 4 and ph 3) on the susceptibility of the larvae of neodiprion sertifer to its nuclear polyhedrosis virus. scots pines growing in a subarctic area with low ambient pollution levels were irrigated with simulated acid rain during two summers. neodiprion larvae fed with foliage from the experimental trees were infected with a dilute virus suspension. the acid treatment of host trees had a significant e ...199022160113
[Design of the composition of baculovirus agents].The ability of 30 compounds to protect infection bacteria and baculoviruses from the damaging effect of ultraviolet (UV) irradiation was investigated. For this B. thuringiensis var. israelensis spores and gypsy moth (Lymantria dispar L.) nuclear polyhedrosis virus were mixed with different components and exposed to UV irradiation at 0.25 WI cm2 for 60 min. Then spore viability and viral pathogenicity were studied in third instar gypsy moth larvae. The composition comprising sodium alginate, albu ...201122171478
[the search of mirna genes in bombyx mori nuclear polyhedrosis virus genomes regions complementary to the latest genes].the search of mirna genes in bombyx mori nuclear polyhedrosis virus genome region complementary to very late genes has been carried out. the search mirna algorithm in silico was developed by us. it was shown that npv b. mori genome region containing orf4 gene complementary to ph gene encodes the potential mirna. npv b. mori genome region containing p74 gene complementary to p10 gene encodes mature mirna and potential mirna. the genome region containing orf1629 encodes two small non-coding rnas c ...201122276429
development of a novel baculovirus titration method using the enzyme-linked immunosorbent spot (elispot) assay.the baculovirus expression vector system (bevs) is one of the most powerful methods for production of recombinant proteins for research or commercial purposes. titration of viable virus in insect cell culture is often required when bevs is used for basic research or bioprocessing. an enzyme-linked immunosorbent spot (elispot) assay using monoclonal antibodies against the major capsid protein vp39 of both autographa californica nuclear polyhedrosis virus (acmnpv) and bombyx mori nuclear polyhedro ...201223274754
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