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expression and mutational analysis of the baculovirus very late factor 1 (vlf-1) gene.we have examined the expression and function of a gene, vlf-1, of autographa californica nuclear polyhedrosis virus that is known to encode a regulator of very late gene transcription. western blot analysis revealed that vlf-1 is expressed during the late phase of infection, primarily from 15 to 24 h postinfection. vlf-1 localized in the cell nucleus and was also present in the nucleocapsids of virus particles. mapping of vlf-1 mrna by primer extension showed that transcription initiates at a ta ...19989614871
effects of substituting granulin or a granulin-polyhedrin chimera for polyhedrin on virion occlusion and polyhedral morphology in autographa californica multinucleocapsid nuclear polyhedrosis virus.substitution of granulin from the trichoplusia ni granulosis virus (tngv) for polyhedrin of the autographa californica multinucleocapsid nuclear polyhedrosis virus (acmnpv) yielded a few very large (2 to 5 micron) cuboidal inclusions in the cytoplasm and nucleus of infected cells. these polyhedra lacked the beveled edges characteristic of wild-type acmnpv polyhedra, contained fractures, and occluded few virions. placing a nuclear localization signal (krkk) in granulin directed more granulin to t ...19989621097
the in vivo production of spodoptera littoralis nuclear polyhedrosis virus.the in vivo production of the nucleopolyhedrovirus (npv) of the egyptian cotton leafworm spodoptera littoralis was studied experimentally. larvae (7 days old) of 30-50 mg were experimentally infected with a range of npv doses then harvested alive at various times after dosing to determine the effect of dose and incubation time on npv productivity. maximum npv production achieved after 7 days incubation was 1.86 x 10(9) polyhedral inclusion bodies (pibs) per larvae using an inoculum of 1 x 10(4) ...19989628227
identification of a novel lymantria dispar nucleopolyhedrovirus mutant that exhibits abnormal polyhedron formation and virion occlusion.in previous studies on the formation of lymantria dispar nuclear polyhedrosis virus (ldmnpv) few polyhedra (fp) mutants, several polyhedron formation mutants (pfm) were identified that appeared to be unique. these viral mutants are being characterized to investigate the processes of polyhedron formation and virion occlusion. ldmnpv isolate pfm-1 is one of these mutants, and is described in this report. genetic techniques were used to determine if isolate pfm-1 contained a mutation in the polyhed ...19989647698
observations on the presence of the peritrophic membrane in larval trichoplusia ni and its role in limiting baculovirus infection.light microscopical examinations of dissected and stained peritrophic membranes (pms) were conducted to determine the presence or absence of this protective structure in larvae of trichoplusia ni, prior to and through ecdysis. observations of fourth- and fifth-instar larvae of t. ni from two independent rearing colonies showed that pms were present and lined the midgut prior to, during, and immediately after ecdysis in both instars. western blot analysis of insect intestinal mucin (iim), a major ...19989647702
detection of a virus enhancing factor in the spheroid, spindle, and virion of an entomopoxvirus.spheroids, spindles, and virions of an entomopoxvirus (epv) enhanced the infectivity of a nuclear polyhedrosis virus (npv) when they were perorally administered to larvae of the armyworm, pseudaletia separata. spheroids and spindles at the same dose exhibited nearly the same enhancing activity. when the dose of spheroids or spindles was reduced 10 times, the median infectious dose of the npv was increased approximately 100 times. an antiserum against an enhancing factor detected the homologous a ...19989647705
expression of biologically active recombinant porcine gm-csf by baculovirus gene expression system.the full length porcine granulocyte/macrophage colony stimulating factor (gm-csf) cdna, including secretion signal peptide coding region was recloned into baculovirus transfer vector pacym1. the vector was then transfected with autographica californica nuclear polyhedrosis virus (acnpv) dna into sf21ae cells and the recombinant virus acpgm was recovered. recombinant porcine gm-csf (rpgm-csf) was obtained from the serum-free culture medium of tn5 cells infected with the acpgm virus, and was shown ...19989682962
a baculovirus single-stranded dna binding protein, lef-3, mediates the nuclear localization of the putative helicase p143.the intracellular localization of the baculovirus autographa californica nuclear polyhedrosis virus p143 gene product (p143) was investigated by immunofluoresence staining of infected and transfected cells. as expected for a protein essential for viral dna replication, under these conditions, p143 was localized to the nucleus. however, when a plasmid directing the synthesis of p143 from its own promoter was co-transfected with a plasmid expressing ie-1, p143 was found in the cytoplasm. cotransfe ...19989683569
characterization of a baculovirus-encoded rna 5'-triphosphatase.autographa californica nuclear polyhedrosis virus (acnpv) encodes a 168-amino-acid polypeptide that contains the signature motif of the superfamily of protein phosphatases that act via a covalent cysteinyl phosphate intermediate. the sequence of the acnpv phosphatase is similar to that of the rna triphosphatase domain of the metazoan cellular mrna capping enzyme. here, we show that the purified recombinant acnpv protein is an rna 5'-triphosphatase that hydrolyzes the gamma-phosphate of triphosph ...19989696798
control of baculovirus polyhedrin gene expression by very late factor 1.vlf-1 is a baculovirus gene that regulates very late gene expression (j. r. mclachlin and l. k. miller, j. virol., 68, 7746-7756, 1994) and also plays a crucial role in the replication of the budded form of autographa californica nuclear polyhedrosis virus (acmnpv) (s. yang and l. k. miller, "expression and mutational analysis of the baculovirus very late factor 1 (vlf-1) gene." virology, 245, 99-109, 1998). to examine the influence of vlf-1 expression on baculovirus infection, we constructed re ...19989705262
a protein tyrosine phosphatase-like protein from baculovirus has rna 5'-triphosphatase and diphosphatase activities.the superfamily of protein tyrosine phosphatases (ptps) includes at least one enzyme with an rna substrate. we recently showed that the rna triphosphatase domain of the caenorhabditis elegans mrna capping enzyme is related to the ptp enzyme family by sequence similarity and mechanism. the ptp most similar in sequence to the capping enzyme triphosphatase is bvp, a dual-specificity ptp encoded by the autographa californica nuclear polyhedrosis virus. although bvp previously has been shown to have ...19989707557
interactions between a nosema sp. (microspora: nosematidae) and nuclear polyhedrosis virus infecting the gypsy moth, lymantria dispar (lepidoptera: lymantriidae).simultaneous and sequential per os inoculations of gypsy moth larvae with the lymantria dispar nuclear polyhedrosis virus (ldnpv) and a nosema sp. from portugal demonstrated that the interaction of two pathogens during coinfection was variable, ranging from synergistic to antagonistic. susceptibility of gypsy moth larvae to viral infection was unaffected by simultaneous and subsequent microsporidian infection. this resulted from the comparatively slow pathogenesis of the microsporidium when comp ...19989709015
a virus-encoded rna polymerase purified from baculovirus-infected cells.a dna-dependent rna polymerase was purified to homogeneity, starting from insect cells infected with the baculovirus autographa californica nuclear polyhedrosis virus (acnpv). the purified polymerase supported accurate and specific transcription from late and very late promoters but was not active on viral early promoters. thus, promoter recognition is an integral function of the purified enzyme. the purified rna polymerase was composed of only four equimolar subunits, which makes it the simples ...19989733837
colourimetric pcr-based detection of monodon baculovirus in whole penaeus monodon postlarvae.the development of a nested polymerase chain reaction (pcr) assay is described to detect low concentrations of monodon baculovirus (mbv) dna from total penaeus monodon postlarval dna. a modified dna extraction procedure was also developed to circumvent problems associated with co-purification of pcr inhibitors in total dna extracted from whole postlarvae. this method involved mechanical disruption of frozen prawn material immediately followed by phenol extraction at high temperature. an assessme ...19989763125
persistent baculovirus infection results from deletion of the apoptotic suppressor gene p35.infection with the wild-type baculovirus autographa californica multiple nuclear polyhedrosis virus (acmnpv) results in complete death of spodoptera frugiperda (sf) cells. however, infection of sf cells with acmnpv carrying a mutation or deletion of the apoptotic suppressor gene p35 allowed the cloning of surviving sf cells that harbored persistent viral genomes. persistent infection established with the virus with p35 mutated or deleted was blocked by stable transfection of p35 in the host geno ...19989765462
the early baculovirus he65 promoter: on the mechanism of transcriptional activation by ie1.we have initiated studies on the mechanism of early transcriptional activation of the early he65 promoter during infection with autographa californica multicapsid nuclear polyhedrosis virus. this analysis is based on a comparison of the sequences required for he65 promoter activation with those sequences that support specific protein binding. the he65 promoter is located immediately downstream of the homologous region (hr) 4a. the sequences of hr4a are characterized by two imperfect palindromes ...19989791025
guanylyltransferase activity of the lef-4 subunit of baculovirus rna polymerase.the baculovirus autographa californica nuclear polyhedrosis virus encodes a dna-dependent rna polymerase that transcribes viral late genes. this polymerase is composed of four equimolar subunits, lef-4, lef-8, lef-9, and p47. here we present data indicating that the lef-4 subunit of rna polymerase is a guanylyltransferase. incubation of rna polymerase in the presence of divalent cation and radiolabeled gtp resulted in the formation of a covalent enzyme-guanylate complex that comigrated with the ...19989811738
the lef-4 subunit of baculovirus rna polymerase has rna 5'-triphosphatase and atpase activities.the baculovirus autographa californica nuclear polyhedrosis virus encodes a dna-dependent rna polymerase that is required for transcription of viral late genes. this polymerase is composed of four equimolar subunits, lef-8, lef-4, lef-9, and p47. the lef-4 subunit has guanylyltransferase activity, suggesting that baculoviruses may encode a full complement of capping enzymes. here we show that lef-4 is a bifunctional enzyme that hydrolyzes the gamma phosphates of triphosphate-terminated rna and a ...19989811739
rna 5'-triphosphatase, nucleoside triphosphatase, and guanylyltransferase activities of baculovirus lef-4 protein.autographa californica nuclear polyhedrosis virus late and very late mrnas are transcribed by an rna polymerase consisting of four virus-encoded polypeptides: lef-8, lef-9, lef-4, and p47. the 464-amino-acid lef-4 subunit contains the signature motifs of gtp:rna guanylyltransferases (capping enzymes). here, we show that the purified recombinant lef-4 protein catalyzes two reactions involved in rna cap formation. lef-4 is an rna 5'-triphosphatase that hydrolyzes the gamma phosphate of triphosphat ...19989811740
nineteen baculovirus open reading frames, including lef-12, support late gene expression.a set of 18 plasmid subclones of the autographa californica nuclear polyhedrosis virus genome, each containing an identified late expression factor gene (lef), supports expression from a late viral promoter in transient expression assays in the sf-21 cell line derived from spodoptera frugiperda. we have constructed a further set of plasmids in which each lef open reading frame (orf) is controlled by the drosophila melanogaster heat shock protein 70 (hsp70) promoter and epitope tagged. failure of ...19989811761
the potential role of a late gene expression factor, lef2, from bombyx mori nuclear polyhedrosis virus in very late gene transcription and dna replication.several late gene expression factors (lefs) have been implicated in fostering high levels of transcription from the very late gene promoters of polyhedrin and p10 from baculoviruses. we cloned and characterized from bombyx mori nuclear polyhedrosis virus a late gene expression factor (bmlef2) that encodes a 209-amino-acid protein harboring a cys-rich c-terminal domain. the temporal transcription profiles of lef2 revealed a 1.2-kb transcript in both delayed early and late periods after virus infe ...19989813207
a mutational analysis of the baculovirus inhibitor of apoptosis op-iap.a family of antiapoptotic regulators known as inhibitors of apoptosis (iaps) was initially identified and functionally described in baculoviruses, and iap homologues are now known in insects, birds, and mammals. baculovirus and drosophila iaps inhibit apoptosis induced by drosophila proapoptotic proteins reaper, hid, and grim and physically interact with them through their baculovirus iap repeat (bir) region. here we examined the functional importance of bir and ring finger motifs of orgyia pseu ...19989852042
expression and cellular distribution of baculovirus-expressed bovine herpesvirus 1 (bhv-1) glycoprotein d (gd) sequences.glycoprotein d (gd) of bovine herpesvirus 1 (bhv-1), a homolog of herpes simplex virus gd, represents a major component of the viral envelope and is a dominant immunogen. to study the antigenic properties of the different regions of gd, we have expressed the full-length gd encoding gene and overlapping fragments spanning various regions of the gd open reading frame in a baculovirus (autographa californica nuclear polyhedrosis virus)--insect cell (spodoptera frugiperda, sf-9) system. maximum leve ...19989856100
in vitro transcription of pe38/polyhedrin hybrid promoters reveals sequences essential for recognition by the baculovirus-induced rna polymerase and for the strength of very late viral promoters.in vitro transcription was used to analyze the promoter specificity of the alpha-amanitin-resistant rna polymerase that is induced late during infection of autographa californica multicapsid nuclear polyhedrosis virus. by modifying the preparation of crude nuclear extracts, we have established an assay that permits differentiation between weak late and strong very late viral promoters. the virus-induced rna polymerase initiates at a taag sequence motif in both late and very late promoters. based ...19989525621
species-specificity in endoplasmic reticulum signal peptide utilization revealed by proteins from trypanosoma brucei and leishmania.n-terminal signal peptides direct secretory and most membrane proteins into the exocytic pathway at the endoplasmic reticulum. signal sequences can function across kingdoms. however, our attempts at translocating variant surface glycoprotein (vsg) 117, vsg mvat7, vsg 221 and bip from trypanosoma brucei and gp63 from leishmania chagasi into canine pancreas microsomes failed. on replacing the signal peptide of vsg 117 with that from yeast prepro-alpha-mating factor (ppalphamf) the chimaeric protei ...19989531493
development and validation of a competitive enzyme-linked immunosorbent assay for detection of type a influenza antibodies in avian sera.serologic screening of avian sera for group-specific antibodies to type a influenza is currently accomplished by using the avian influenza (ai) agar gel immunodiffusion (agid) test. a competitive enzyme-linked immunosorbent assay (celisa) was developed using a baculovirus vector, autographa californica nuclear polyhedrosis virus, expressing the nucleoprotein (np) gene of a/ann arbor/6/60 influenza virus. the recombinant np was obtained by inoculation of spodoptera frugiperda (sf9) insect cells o ...19989533078
characterization of a nuclear polyhedrosis virus of spodoptera frugiperda from argentina. 19989538035
new isolate of macosoma neustria nuclear polyhedrosis virus in latvia. 19989538036
in vitro and in vivo host range of anticarsia gemmatalis multiple nuclear polyhedrosis virus.a clone of the wild type (wt) anticarsia gemmatalis multiple nuclear polyhedrosis virus agmnpv, derived from a geographical isolate (hondrina, brazil) and designated agmnpv-cl4-3a1, was used to determine the host range of this virus in six established lepidopteran cell lines: anticarsia gemmatalis (bcirl-ag-am1), helicoverpa zea (bcirl-hz-am1), heliothis virescens (bcirl-hv-am1), helicoverpa armigera (bcirl-ha-am1), trichoplusia ni (tn-cl1), bombyx mori (bmn), and a coleopteran cell line anthono ...19989542640
removal of 106 amino acids from the n-terminus of udp-glcnac: alpha-3-d-mannoside beta-1,2-n-acetylglucosaminyltransferase i does not inactivate the enzyme.udp-glcnac: alpha-3-d-mannoside beta-1,2-n-acetylglucosaminyltransferase i (gnti, ec 2.4.1.101) plays an essential role in the conversion of oligomannose to complex and hybrid n-glycans. rabbit gnti is 447 residues long and has a short four-residue n-terminal cytoplasmic tail, a 25-residue putative signal-anchor hydrophobic domain, a stem region of undetermined length and a large c-terminal catalytic domain, a structure typical of all glycosyltransferases cloned to date. comparison of the amino ...19989557881
role of baculovirus ie2 and its ring finger in cell cycle arrest.the ie2 gene of autographa californica nuclear polyhedrosis virus (acmnpv) is known to transactivate transient expression from viral promoters in a host cell-specific manner. we report that transfection of spodoptera frugiperda (sf-21) cells with ie2 was sufficient to arrest the cell cycle, resulting in the accumulation of enlarged cells with abnormally high dna contents. by 72 h posttransfection, more than 50% of ie2-transfected cells had dna contents greater than 4n. there was no evidence of m ...19989420274
in vivo and in vitro analysis of baculovirus ie-2 mutants.upon transient expression in cell culture, the ie-2 gene of autographa californica nuclear polyhedrosis virus (acmnpv) displays three functions: trans activation of viral promoters, direct or indirect stimulation of virus origin-specific dna replication, and arrest of the cell cycle. the ability of ie2 to trans stimulate dna replication and coupled late gene expression is observed in a cell line derived from spodoptera frugiperda but not in a cell line derived from trichoplusia ni. this finding ...19999971831
activation of baculovirus very late promoters by interaction with very late factor 1.very late factor 1 (vlf-1) of autographa californica multicapsid nuclear polyhedrosis virus (acmnpv) activates the transcription of two genes, polyhedrin (polh) and p10, during the final, occlusion-specific phase of infection. using transient expression assays responsive to vlf-1, we identified linker scan mutations in the polh and p10 promoters which abolished or weakened the ability of the promoters to respond to stimulation by vlf-1. these mutations were located between the transcriptional an ...199910074194
development and use of a pcr assay for detection of the reproductive virus in wild populations of helicoverpa zea (lepidoptera: noctuidae)helicoverpa zea reproductive virus (hzrv) is a nonoccluded bacilliform virus that affects both female and male moths of the corn earworm h. zea. in order to study the biology and host range of hzrv, a bioassay was previously developed to detect the presence of this virus in infected insects. a drawback of this bioassay is that it is time consuming and requires more than a month to complete. here we describe the development of a polymerase chain reaction (pcr) assay for the rapid detection of hzr ...19999878296
baculovirus p33 binds human p53 and enhances p53-mediated apoptosis.in vertebrates, p53 participates in numerous biological processes including cell cycle regulation, apoptosis, differentiation, and oncogenic transformation. when insect sf-21 cells were infected with a recombinant of the baculovirus autographa californica nuclear polyhedrosis virus (acmnpv) overexpressing human p53, p53 formed a stable complex with the product of the acmnpv orf92, a novel protein p33. the interaction between p53 and p33 was further confirmed by immunoprecipitation studies. when ...19999882325
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
an analysis of the role of the target membrane on the gp64-induced fusion pore.influenza hemagglutinin (ha) and gp64 of the baculovirus autographa californica multicapsid nuclear polyhedrosis virus induce strikingly different initial fusion pores when mediating fusion between host cells that express these fusion proteins and target cells (plonsky and zimmerberg, 1996; spruce et al., 1989, 1991; zimmerberg et al., 1994). however, in these experiments, variations in host and target membranes confounded the analysis of the role that major components of the fusion reaction pla ...19999887319
human interleukin-2 production in insect (trichoplusia ni) larvae: effects and partial control of proteolysis.many eukaryotic proteins have been successfully expressed in insect cells infected with a baculovirus in which the foreign gene has been placed under the control of a viral promoter. this system can be costly at large scale due to the quality of virus stock, problems of oxygen transfer, and severity of large-scale contamination. to circumvent this problem, we have investigated the expression of a foreign protein, human interleukin-2 (il-2), in insect larvae, trichoplusia ni, infected with the ba ...199910099527
purification of human interleukin-2 fusion protein produced in insect larvae is facilitated by fusion with green fluorescent protein and metal affinity ligand.the fusion protein of green fluorescent protein (gfp) and human interleukin-2 (hil-2) was produced in insect trichoplusia ni larvae infected with recombinant baculovirus derived from the autographa californica nuclear polyhedrosis virus (acnpv). this fusion protein was composed of a metal ion binding site (his)6 for rapid one-step purification using immobilized metal affinity chromatography (imac), uv-optimized gfp (gfpuv), enterokinase cleavage site for recovering hil-2 from purified fusion pro ...199910194406
expression and purification of human interleukin-2 simplified as a fusion with green fluorescent protein in suspended sf-9 insect cells.a fusion protein of human interleukin-2 (hil-2) and green fluorescent protein (gfp) was expressed in insect sf-9 cells infected with recombinant baculovirus derived from the autographa californica nuclear polyhedrosis virus (acnpv). this fusion protein was comprised of a histidine affinity ligand for simplified purification using immobilized metal affinity chromatography (imac), uv-optimized gfp (gfpuv) as a marker, an enterokinase cleavage site for recovery of hil-2 from the fusion, and the mod ...199910201111
use of the baculovirus system to assemble polyomavirus capsid-like particles with different polyomavirus structural proteins: analysis of the recombinant assembled capsid-like particles.the genes encoding the structural proteins (vp1, vp2 and vp3) of murine polyomavirus were cloned into the p2bac dual multiple cloning site vector, individually or jointly, and the corresponding proteins were expressed in spodoptera frugiperda (sf9) insect cells by cotransfecting sf9 cells with the constructed vector and the linear dna of autographa californica multiple nuclear polyhedrosis virus (acmnpv). recombinant capsid-like particles could be purified 5 days post-infection from sf9 cells in ...199910211971
a field release of genetically engineered gypsy moth (lymantria dispar l.) nuclear polyhedrosis virus (ldnpv).the gypsy moth (lymantria dispar l.) nuclear polyhedrosis virus was genetically engineered for nonpersistence by removal of the gene coding for polyhedrin production and stabilized using a coocclusion process. a beta-galactosidase marker gene was inserted into the genetically engineered virus (ldgev) so that infected larvae could be tested for its presence using a colorimetric assay. in 1993, ldgev-infected gypsy moths were released in a forested plot in massachusetts to test for spread and pers ...199910222179
autographa californica nuclear polyhedrosis virus dna polymerase: measurements of processivity and strand displacement.the dna polymerase (dnapol) of autographa californica nuclear polyhedrosis virus was purified to homogeneity from recombinant baculovirus-infected cells. dnapol was active in polymerase assays on singly primed m13 template, and full-length replicative form ii product was synthesized at equimolar ratios of enzyme to template. the purified recombinant dnapol was shown to be processive by template challenge assay. furthermore, dnapol was able to incorporate hundreds of nucleotides on an oligo(dt)-p ...199910233952
human pir1 of the protein-tyrosine phosphatase superfamily has rna 5'-triphosphatase and diphosphatase activities.a human cdna was isolated encoding a protein with significant sequence similarity (41% identity) to the bvp rna 5'-phosphatase from the autographa californica nuclear polyhedrosis virus. this protein is a member of the protein-tyrosine phosphatase (ptp) superfamily and is identical to pir1, shown by yuan et al. (yuan, y., da-ming, l., and sun, h. (1998) j. biol. chem. 272, 20347-20353) to be a nuclear protein that can associate with rna or ribonucleoprotein complexes. we demonstrate that pir1 re ...199910347225
temperature dependent characteristics of a recombinant infectious hematopoietic necrosis virus glycoprotein produced in insect cells.a recombinant infectious hematopoietic necrosis virus (ihnv) glycoprotein (g protein) was produced in insect cells using a baculovirus vector (autographa californica nuclear polyhedrosis virus). characteristics of this protein were evaluated in relation to native viral g protein. a full-length (1.6 kb) cdna copy of the glycoprotein gene of ihnv was inserted into the baculovirus vector under control of the polyhedrin promoter. high levels of g protein (approximately 0.5 microgram/1 x 10(5) cells) ...199910349547
generation of a new protein purification matrix by loading ceramic hydroxyapatite with metal ions--demonstration with poly-histidine tagged green fluorescent protein.the gene encoding the green fluorescent protein (gfp) from the jellyfish aequorea victoria, was inserted under transcriptional control of the polyhedrin promoter of the autographa californica nuclear polyhedrosis virus and expressed in the spodoptera frugiperda insect cell line sf9 during viral infection. the baculovirus transfervector pbluebachisb was used for constructing the recombinant baculovirus, so that the green fluorescent protein could be tagged with a poly-histidine tail. this fusion ...199910361722
mitochondrial dna acts as potential promoter of the baculovirus rna polymerase.we have examined whether mitochondrial dna could act as target of the rna polymerase encoded by the baculovirus autographa californica multicapsid nuclear polyhedrosis virus, because the baculovirus late promoters and the control region of host mitochondrial dna show a high degree of sequence similarity. in vitro transcription using mitochondrial dna from spodoptera frugiperda cells and nuclear extracts prepared from baculovirus infected cells demonstrates that mitochondrial dna is recognized by ...199910384964
preparation of recombinant rat interleukin-5 by baculovirus expression system and analysis of its biological activities.rat interleukin-5 (il-5) cdna was subcloned from peritoneal cells collected 4 h after intraperitoneal injection of ascaris suum antigen solution into the immunized rats. cysteine proteinase-deleted (cpd) rat il-5 recombinant virus was constructed by inserting rat il-5 cdna into cpd virus having a deletion in the cysteine proteinase gene of the silkworm bombyx mori nuclear polyhedrosis virus. on infection with the cpd rat il-5 recombinant virus, the silkworm b. mori larvae produced rat il-5 as a ...199910446387
the establishment of new cell lines from pseudaletia unipuncta with differential responses to baculovirus infection.six insect cell lines from pseudaletia unipuncta embryos were established and characterized, and their susceptibility to autographa californica multiple nuclear polyhedrosis virus (acmnpv) infection was investigated. these embryonic p. unipuncta cell lines had characteristics distinct from each other in morphology and growth, and showed differential responses to acmnpv infection. among the six cell lines, two were highly susceptible to virus infection. one of these two cell lines, bti-pu-a7s, pr ...199910476920
the characterization and phylogenetic relationship of the trichoplusia ni single capsid nuclear polyhedrosis virus polyhedrin gene.the polyhedrin gene (polh) was identified from the trichoplusia ni (tni) single capsid nuclear polyhedrosis virus (snpv). an ecori fragment containing the truncated polyhedrin gene was detected by hybridization with an acmnpv expression vector probe; the remaining portion of the gene was amplified by reverse pcr. an open reading frame (orf) of 741 nucleotides (nt), encoding a putative protein of 246 amino acids (a.a) with mr 28,780 da was identified. the 5'-noncoding region contained the putativ ...199910499452
[comparative study of aerosol and oral methods of infection of lympantria dispar l. caterpillars with nuclear polyhedrosis virus].the aerosol and oral routes of infection of gypsy moth larvae with nuclear polyhedrosis virus are compared. the virus in aerosol retains its biological activity. the virus output/expenditure ratio is virtually the same in the studied routes of infection. aerosol method of inoculation saves 30% components of media and is 6-8 times less labor consuming. this method permits complete automation of infection of larvae, thus essentially improving the efficacy of baculovirus production.199910500990
expression of the fusion protein recombinant human granulocyte-macrophage colony stimulating factor and leukemia inhibitory factor in a baculovirus vector system.a fusion gene coding human granulocyte-macrophage colony stimulating factor (gm-csf) and leukemia inhibitory factor (lif) cdnas was inserted into the transfer vector psxivvi+ x3 with the control of syn and xiv promoters. the sf9 cells (spodoptera frugiperda) were co-transfected with the recombinant plasmid and tnnpv dna (trichoplusia ni nuclear polyhedrosis virus dna). the fusion protein recombinant human granulocyte-macrophage colony stimulating factor (gm-csf) and leukemia inhibitory factor (l ...199910510487
detection of baculovirus-infected insect cells by flow cytometric side-scatter analyses.the baculovirus expression vector system (bevs), utilizing the autographa californica nuclear polyhedrosis virus (acnpv), has turned out to be an attractive alternative for high-level expression (<600 mg/l) of recombinant proteins. however, there is a shortage of reliable methods for monitoring the infection process in situations where marker proteins cannot be used.199910520205
expression of genes coding for animal virus glycoproteins in heterologous systems.the outermost layers of animal viruses are usually composed of glycoproteins. they are responsible not only for the entrance of viruses into, and release from host cells but also for the initial interaction of a viral particle with immunological defense of the host. it is therefore not surprising that many laboratories devote a lot of effort to study viral glycoproteins at the molecular level. very often such studies are possible only after the introduction of a glycoprotein gene into a heterolo ...199910547034
baculovirus-based genetic screen for antiapoptotic genes identifies a novel iap.the prototype baculovirus, autographa californica multiple nuclear polyhedrosis virus (acmnpv) expresses p35, a potent anti cell-death gene that promotes the propagation of the virus by blocking host cell apoptosis. infection of insect sf-21 cells with acmnpv lacking p35 induces apoptosis. we have used this pro-apoptotic property of the p35 null virus to screen for genes encoding inhibitors of apoptosis that rescue cells infected with the p35 defective virus. we report here the identification of ...199910593985
[biosynthesis of a single peptide chain containing human chorionic gonadotropin beta and c3d of complement].in view of the strong immunity-enhancing function of hel-c3d3 designed by dr. paul w. dempsey, we made our efforts to produce a similar recombinant protein of hcg beta. with polymerase chain reaction, we introduced a bam hi restriction site into the 3' terminal of hcg beta cdna. the new cdna and its terminal's correctness has been confirmed by sequencing. then we have it covalently attached to the c3d3 cdna at the pre-designed bam hi/bgl ii site. having the chimeric dna correctly cloned into the ...199912548792
[expression of schistosoma japonicun fatty acid binding protein gene in silkworm cells and larvae].to express the fatty acid binding protein (sj14fabp) gene of schistosoma japonicun in the silkworm cells and larvae.199912563767
a baculovirus enhancin alters the permeability of a mucosal midgut peritrophic matrix from lepidopteran larvae.the peritrophic matrix (pm) in lepidopterous larvae may function as a defensive barrier against ingested viral pathogens. pms isolated from trichoplusia ni and pseudaletia unipuncta larvae, were treated with a baculovirus-encoded metalloprotease (enhancin) from trichoplusia ni granulosis virus (tngv) and their in vitro permeability to blue dextran and fluorescent-labelled autographa californica nuclear polyhedrosis virus (acmnpv) was determined using a dual chamber permeability apparatus. incuba ...199912770384
assessment of the application of baculoviruses for control of lepidoptera.baculoviruses, among other insect viruses, are regarded as safe and selective bioinsecticides, restricted to invertebrates. they have been used worldwide against many insect pests, mainly lepidoptera. their application as microbial pesticides, however, has not met their potential to control pests in crops, forests, and pastures, with the exception of the nuclear polyhedrosis virus of the soybean caterpillar (anticarsia gemmatalis), which is used on approximately 1 million ha annually in brazil. ...199915012374
expression of a gene for single-chain antibody to carcinoembryonic antigen in bombyx mori cells and its larvae.recombinant bm-bacscfv virus which contains a cdna encoding anti-cea single-chain antibody was generated by cotransfection into bm n cells with the transfer plasmid pbacpak-(his)(6)-scfv and the modified bombyx mori nuclear polyhedrosis virus bm-bacpak genomic dna. bombyx mori cells and larvae were infected by the recombinant virus, respectively. the anti-cea single-chain antibody were expressed both in bombyx mori cells and larvae, the former accounted for 6% of the total cell protein, the late ...199912110934
sequence analysis of sod gene of bombyx mori nuclear polyhedrosis virus and its expression in e.coli.superoxide dismutases scavenge superoxide radicals and protect cells from oxidative stress. sod gene of bombyx mori nuclear polyhedrosis virus has been cloned by pcr, and is expressed in e.coli with the activity of sod being 576.13 u/ml. dna sequence analysis shows that sod gene of bmnpv encodes 151 amino acids. the homology between bmnpv and acnpv of the nucleotide sequences of sod gene is 97.2%, and 56% between bmnpv and human sod1.199912114994
efficient secretion of proteins expressed from insect cells directed by pam signal peptide.signal and leading peptide sequences of rat pam was inserted into the baculovirus transfer vector, and secretion expression plasmids pbacpag2 and pbacpai for the fusion gene pabc-hgrf and pabc-igf-i were constructed, respectively. by cotransfection with linear genomic dna of modified autographa californica nuclear polyhedrosis virus (bacpak6) and homologous recombination, the recombinant acnpv, bacpag and bacpai, were obtained and identified. fusion proteins pabc-hgrf and pabc-igf-i were secrete ...199912142920
visualization of a recombinant gene protein in the baculovirus expression vector system using confocal scanning laser microscopy.expression of the recombinant protein beta-galactosidase in the spodoptera frugiperda sf-9 insect cell line infected by the autographa californica nuclear polyhedrosis virus expressing beta-galactosidase (acnpv-betagal) was visualized using confocal scanning laser microscopy with fluorescent staining of both the recombinant protein and the cell nucleus. the average size of the insect cells and the intracellular dna concentration both increased markedly, respectively reading 3.8- and 2.3-fold the ...199916232550
[application of nuclear polyhedrosis virus of insects in parasitology]. 200012567649
diagnosis of penaeus monodon-type baculovirus by pcr and by elisa of occlusion bodies.the black tiger prawn penaeus monodon is a valuable aquaculture product in taiwan. two specific diagnostic methods were established for p. monodon-type baculovirus, one using polymerase chain reaction (pcr) technology and the other enzyme-linked immunosorbent assay (elisa) technology. monodon-type baculovirus (mbv) was purified by sucrose gradient centrifugation from occlusion bodies of mbv-infected postlarvae of p. monodon. mbv dna was subsequently purified from the occlusion bodies and its pre ...200010782342
differential activity of two non-hr origins during replication of the baculovirus autographa californica nuclear polyhedrosis virus genome.the identification of potential baculovirus origins of replication (ori) has involved the generation and characterization of defective interfering particles that contain major genomic deletions yet retain their capability to replicate by testing the replication ability of transiently transfected plasmids carrying viral sequences in infected cells. so far, there has not been any evidence to demonstrate the actual utilization of these putative origins in autographa californica multinucleocapsid nu ...200010799593
the autographa californica nuclear polyhedrosis virus p143 gene encodes a dna helicase.the p143 protein of autographa californica nuclear polyhedrosis virus is essential for replication of viral dna. to determine the function of p143, the protein was purified to near homogeneity from recombinant baculovirus-infected cells that overexpress p143. atpase activity copurified with p143 protein during purification and also during gel filtration at a high salt concentration. the atpase activity did not require the presence of single-stranded dna, but was stimulated fourfold by the additi ...200010799604
the expression and biologic effects of ovine interleukin-4 on t and b cell proliferation.using the reverse-transcriptase polymerase chain reaction (rt-pcr), cdna encoding ovine (ov) interleukin-4 (ovil-4) was generated from mitogen-stimulated peripheral blood mononuclear cells (pbmc). two identical clones generated from separate rt-pcr reactions differed from a published ovil-4 sequence, although they had a high degree of identity with the bovine and human homologs. we show by sequence analysis that the ovil-4 cdna retained the four alpha-helix structure and disulfide bonds identifi ...200010805377
pathogen-driven outbreaks in forest defoliators revisited: building models from experimental data.models of outbreaks in forest-defoliating insects are typically built from a priori considerations and tested only with long time series of abundances. we instead present a model built from experimental data on the gypsy moth and its nuclear polyhedrosis virus, which has been extensively tested with epidemic data. these data have identified key details of the gypsy moth-virus interaction that are missing from earlier models, including seasonality in host reproduction, delays between host infecti ...200010856195
expression of highly controllable genes in insect cells using a modified tetracycline-regulated gene expression system.a modified tetracycline-responsive expression system (tres) for use in insect cells was developed. the tres contains two components: one encodes a tetracycline-controllable transactivator (tta) and the other contains a tet operator dna sequence to drive the luciferase gene. our results show that the human cytomegalovirus (cmv) promoter, an essential part for strong tta expression in mammalian system, was not functional in insect cells. thus further modifications were required. functional tta was ...200010862988
identification and characterization of the trichoplusia ni single capsid nuclear polyhedrosis virus p10 gene.the p10 gene was identified and characterized from the trichoplusia ni single capsid nuclear polyhedrosis virus (tnisnpv). the p10 open reading frame (orf) sequence was identified following sequencing of the ends of the ecori-g clone. subsequent sequencing of an ecori-smai subclone identified the entire p10 and a portion of a p26 homologue. the p10 orf of 264 basepairs (bps), encoded a predicted protein of 88 amino acids (aas) with mr 9527 da. the putative late transcription initiation motif (ta ...200010872882
enhancement in activity of homologous and heterologous baculoviruses infectious to beet armyworm (lepidoptera: noctuidae) by an optical brightener.the nuclear polyhedrosis virus (npv) from the beet armyworm, spodoptera exigua (hübner) (semnpv), was the most active virus tested against the beet armyworm (lc50 = 4.1 pibs/mm2), followed by nuclear polyhedrosis viruses from the alfalfa looper, autographa californica (speyer) (acmnpv; lc50 = 92.6 pibs/mm2), and the celery looper, anagrapha falcifera (kirby) (afmnpv; lc50 = 195.7 pibs/mm2). in the case of the nuclear polyhedrosis virus from the bollworm, helicoverpa armigera (hübner), lc50s coul ...200010902301
baculoviral display of functional scfv and synthetic igg-binding domains.viral vectors displaying specific ligand binding moities such as scfv fragments or intact antibodies hold promise for the development of targeted gene therapy vectors. in this report we describe baculoviral vectors displaying either functional scfv fragments or the synthetic z/zz igg binding domain derived from protein a. display on the baculovirus surface was achieved via fusion of the scfv fragment or z/zz domain to the n-terminus of gp64, the major envelope protein of the autographa californi ...200010944446
peroral infection of nuclear polyhedrosis virus budded particles in the host, bombyx mori l., enabled by an optical brightener, tinopal unpa-gx.perorally inoculated budded particles of a nuclear polyhedrosis virus was used to infect bombyx mori (bmnpv) (lepidoptera; bombycidae), aided by an optical brightener, tinopal unpa-gx (tinopal). bmnpv budded particles not occluded in the occlusion body do not infect successfully the host, b. mori, when administered perorally. it was found that feeding the host tinopal enabled perorally delivered bmnpv budded particles to infect the host. b. mori larvae ingesting bmnpv budded particles (1.3 x 10( ...200010960702
presentation of antigenic sites from foot-and-mouth disease virus on the surface of baculovirus and in the membrane of infected cells.we describe the construction of recombinant baculoviruses displaying on their surface and in the membrane of infected cells the small, immunodominant antigenic site (site a) or the large polyprotein (p1) coding for the four structural proteins of foot-and-mouth disease virus (fmdv). the coding sequences were inserted in the amino-terminus of gp64, the major glycoprotein of the baculovirus autographa californica nuclear polyhedrosis virus (acnpv). following infection of insect cells with the reco ...200011043943
baculovirus-mediated gene transfer into pancreatic islet cells.baculovirus transduction is a gene transfer method that uses a moth cell virus for mammalian cells in culture, which results in a high-level prolonged expression. here we demonstrate that recombinant baculoviruses can serve as efficient gene transfer vehicles for delivering foreign genes driven by mammalian promoters into human and mouse pancreatic islet cells. existing methods, such as various transfection and electroporation techniques, either suffer from low efficiency or cause extensive memb ...200011117998
effect of two granulosis viruses on the activity of the gypsy moth (lepidoptera: lymantriidae) nuclear polyhedrosis virus.two granulosis viruses (gv) were tested as enhancers for the gypsy moth nuclear polyhedrosis virus (ldmnpv). helicoverpa armigera (hübner) cv (hagv) had no detrimental effect upon larval growth and development, but in combination with ldmnpv it reduced both the lc50 and the lt50 for the npv. in addition, the combination also adversely affected the growth and development of gypsy moth larvae. the lc50 of ldmnpv was reduced by as much as 300-fold (hagv at 10(-2) dilution) and the lt50 was reduced ...200011142292
enhancing the efficacy and persistency of spodoptera litura (fab.) nuclear polyhedrosis virus using uv irradiation protectants.to enhance the field persistency of s. litura nuclear polyhedrosis virus (slnpv), three chemicals viz. cupric ammonium nitrate, tinopal and cupric sulphate were tried as protectants (0.01 mg/ml) against natural sunlight (uv) irradiation. on exposure for 8 hr and subsequent bioassaying (diet surface treatment), it was found that cupric sulphate protected the polyhedrosis inclusion bodies (pibs), recording 95.56% mortality which was statistically at par with unexposed pibs recording 97.78% mortali ...200011395967
[mutagenic analysis on the polyhedrin gene (polh) of bombyx mori nuclear polyhedrosis virus (bmnpv)].in our early studies, the abnormal shape of the polyhedra of bombyx mori nuclear polyhedrosis virus (bmnpv) induced by chemical mutagens of mmc. 9-aa and ems occurred, and the genome of the mutated bmnpv obtained from the successive test had some change in the restriction endonuclease partners of ecori, bglii and bamhi. the present studies showed that the arrangement of the crystal lattice of the polyhedrin was disorderly, and the sds-page electropherogram of the polyhedrin depicted distinct cha ...200011209691
inactivation analysis of hcnpv cysteine protease gene and chitinase gene.the fragment of hyphantria cunea nuclear polyhedrosis virus (hcnpv), which contains cysteine protease gene(cp) and chitinase gene (chia),was inserted into plasmid pcrii to construct transfer vector phccvdel. recombinant transfer vector phccvpolh was constructed by inserting the fragment of hcnpv polyhedrin gene (polh) into the ecori site of the phccvdel. by contransfection of the phccvpolh and hcnpv-ptth(+) dna, which contained bombyx mori prothoracicotropic hormone gene (ptth) into spim cells, ...200012098800
cloning and sequence analysis of the chitinase gene of spodoptera litura nuclear polyhedrosis virus.using acmnpv chia-containing fragment as a probe, the chitinase gene of spodoptera litura nuclear polyhedrosis virus (spltnpv) was localized in two contiguous fragments, xbai 5.1 kb and 2.1 kb, and the intact spltnpv chia gene was obtained by cloning and sequencing those two fragments. the complete open reading frame of the gene was 1 695 nucleotide long, encoding a putative protein of 564 amino acids with molecular weight of 62.9 kd. sequence analysis further revealed that the 5' noncoding regi ...200012058207
[a viral infection in the neck glandula of larvae of dione junio (lepidoptera: nymphalidae)].lepidopteran larvae may be attacked by different viruses, many of which belong to the baculoviridae family. whilst studying the ultrastructure of the neck gland in dione junio larvae we found that in later instars the larvae showed symptoms of attack by two types of virus. the glands were prepared for optical and electron microscopy using sodium cacodylate buffer and standard procedures (0.1m, ph 7.2). the neck gland is composed of two oval internal sacks which communicate with the exterior via ...200011460790
a new continuous cell line from larval hemocytes of spodoptera litura (f.).a new cell line has been established from larval hemocytes of the moth, s. litura (tobacco cut worm). it took 147 days to form a monolayer and one year for the first 17 passages. at present, the culture is at 86th passage level and is designated niv-su-1095. three cell types could be distinguished, viz. plasmatocytes (53%), prohemocytes (36%) and granular hemocytes (11%). the chromosome number was very high, 74% metaphase cells showed more than 100 chromosomes. the cells could be cryopreserved. ...200011411040
[epizootic dynamics of spodoptera litura nuclear polyhedrosi virus].through investigation of tracking infected spodoptera litura larvae, the epizootic dynamics of s. litura nuclear polyhedrosis virus(slnpv) in field was studied with different dosages of slnpv. the results showed that in the range of virus dosages (3.1 x 10(5)-3.1 x 10(8) pibs.ml-1), the initial infection syndrome was observed after 4 days of treatment, and peaked in 5-7 days. the larvae mortality occurred after 5-6 days of treatment, and peaked in 6-8 days. the prevalence peak of host was basica ...200011767686
expression and one-step purification of intracellular human prolactin in insect cells.human prolactin was expressed in insect culture cells by recombinant baculoviruses carrying prolactin gene cdna placed under the transcriptional control of polyhedrin gene promoter of autographa californica nuclear polyhedrosis virus. preliminary results of recombinant human prolactin expression as extracellular as well as intracellular product of baculovirus expression system were presented at the febs meeting in nice, france, in 1999 (abstracts, p. 288). in the present work prolactin was expre ...200111437600
both lymantria dispar nucleopolyhedrovirus enhancin genes contribute to viral potency.enhancins are a group of proteins first identified in granuloviruses (gv) that have the ability to enhance nuclear polyhedrosis virus potency. we had previously identified an enhancin gene (e1) in the lymantria dispar multinucleocapsid nucleopolyhedrovirus (ldmnpv) (d. s. bischoff and j. m. slavicek, j. virol. 71:8133-8140, 1997). inactivation of the e1 gene product within the viral genome lowered viral potency by an average of 2.9-fold. a second enhancin gene (e2) was identified when the entire ...200111507209
identification, sequence analysis, and phylogeny of the immediate early gene 1 of the trichoplusia ni single nucleocapsid polyhedrosis virus.substantial research has been conducted on the immediate early i (ie-1) genes from the prototype baculovirus auographa californica multicapsid nuclear polyhedrosis virus (acmnpv) and the orgyia pseudotsugata multicapsid nuclear polyhedrosis virus (opmnpv). in both cases ie-1 gene products have been implicated in transcriptional activation and repression. in this study an ie-1 homolog was identified from trichoplusia ni single nucleocapsid polyhedrosis virus (tnisnpv). nucleotide sequence analysi ...200111556402
investigation of sequential behavior of carboxyl protease and cysteine protease activities in virus-infected sf-9 insect cell culture by inhibition assay.proteases produced during the culture of spodoptera frugiperda sf-9 cells infected with autographa californica nuclear polyhedrosis virus (acnpv) were assayed with various protease inhibitors. this inhibitory analysis revealed that: (1) carboxyl and cysteine proteases were predominantly produced by the insect cells infected with recombinant acnpv, the gene of which encoded a variant of green fluorescent protein in a portion of the polyhedrin gene of the baculovirus, and (2) the protease activity ...200111601624
effects of deletion and overexpression of the autographa californica nuclear polyhedrosis virus fp25k gene on synthesis of two occlusion-derived virus envelope proteins and their transport into virus-induced intranuclear membranes.partial deletions within autographa californica open reading frame 61 (fp25k) alter the expression and accumulation profile of several viral proteins and the transport of occlusion-derived virus (odv)-e66 to intranuclear membranes during infection (s. c. braunagel et al., j. virol. 73:8559-8570, 1999). here we show the effects of a full deletion and overexpression of fp25k on the transport and expression of two odv envelope proteins, odv-e66 (e66) and odv-e25 (e25). deletion and overexpression o ...200111602724
persistence of an occlusion-negative recombinant nucleopolyhedrovirus in trichoplusia ni indicates high multiplicity of cellular infection.we use data from the serial passage of co-occluded recombinant autographa californica nuclear polyhedrosis virus (acmnpv) to estimate the viral multiplicity of infection of cells within infected insects. co-occlusion, the incorporation of wild-type and mutant virus genomes in the same occlusion body, has been proposed as a strategy to deliver genetically modified viruses as insecticides in a way that contains their spread in the environment. it may also serve as a means whereby naturally occurri ...200111679346
identification of apoptosis-inhibiting gene in leucania separata nuclear polyhedrosis virus.a novel gene lsp40 from leucania separata nuclear polyhedrosis virus (lsnpv) which was homologous to the p35 gene from autographa californica nuclear polyhedrosis virus (acnpv) was localized in the ecorv-5.5 kb fragment of lsnpv genome dna and was sequenced. the open reading frame (orf) of lsp40 was 906 bp long and encoded an approximately 40 kda peptide consisting of 302 amino acid residues. the isp40 shares 80.4% and 70.4% identity of nucleotide and amino acid sequence, respectively, to the ac ...200111765917
functional analysis of helicase gene promoter and homologous region 3 enhancer in bombyx mori nuclear polyhedrosis virus.the promoter of the helicase gene, including 510 bp upstream of atg,was cloned and sequenced, and was found that it had both early and late rna initiation sites. the initiation codon atg was deleted by using point mutation. luciferase gene, as a reporter gene, was fused with the promoter region to construct the plsmid pbm hel 510 luc. when pbm hel 510 luc was transfected into bm-5 and sf-21 cell lines, the helicase gene promoter was recognized by cellular rna polymerase and transactivated by vir ...200112040393
the effects of cations on the activity of the gypsy moth (lepidoptera: lymantriidae) nuclear polyhedrosis virus.fourteen cations were tested at a 1% concentration (wt:wt), as chlorides, for their effects on the biological activity of the gypsy moth, lymantria dispar (l.), nuclear polyhedrosis virus (ldmnpv). cupric chloride was toxic to gypsy moth larvae. ferrous and ferric chloride were inhibitory to larval growth and development as well as to virus activity. strontium chloride was inhibitory to virus activity but had no apparent effects on gypsy moth larvae. six cations had little or no effect on virus ...200111233097
dynamic nuclear localization of the baculovirus proteins ie2 and pe38 during the infection cycle: the promyelocytic leukemia protein colocalizes with ie2.the early gene products ie2 and pe38 of autographa californica multicapsid nuclear polyhedrosis virus localize to distinct nuclear domains after transient expression. here, the nuclear localization pattern and the putative association with cellular proteins have been determined during virus infection to shed light on the functional significance of the nuclear domains. ie2 was always localized to distinct nuclear structures while pe38 was partly present in nuclear dots. confocal imaging indicated ...200111262179
actin rearrangement-inducing factor of baculoviruses is tyrosine phosphorylated and colocalizes to f-actin at the plasma membrane.in previous studies we have identified actin rearrangement-inducing factor 1 as an early gene product of autographa californica multicapsid nuclear polyhedrosis virus that is involved in the remodeling of the actin cytoskeleton. we have constructed viral recombinants with a mutated arif-1 open reading frame that confirm the causal link of arif-1 expression and the actin rearrangement observed as accumulation of f-actin at the plasma membrane at 3 to 7 h postinfection. infection with arif mutant ...200111264366
novel sp family-like transcription factors are present in adult insect cells and are involved in transcription from the polyhedrin gene initiator promoter.we earlier documented the involvement of a cellular factor, polyhedrin (polh) promoter-binding protein, in transcription from the autographa californica nuclear polyhedrosis virus polh gene promoter. sequences upstream of the polh promoter were found to influence polh promoter-driven transcription. analysis of one such region, which could partially compensate for the mutated polh promoter and also activate transcription from the wild-type promoter, revealed a sequence (acsp) containing a caccc m ...200111294840
combination treatment for osteosarcoma with baculoviral vector mediated gene therapy (p53) and chemotherapy (adriamycin).the insect baculovirus autographa californica multiple nuclear polyhedrosis virus (acmnpv) has been evaluated as a vector for gene delivery to human tumor cells. a human osteogenic sarcoma cell line, saos-2, was found to be highly susceptible to infection with a baculoviral vector, with nearly 100% of saos-2 cells being able to express a lacz reporter gene after a brief exposure to the virus at a m.o.i. of 30 pfu/cell. the production of beta-galactosidase protein was 18-times greater than that i ...200111322486
relative effectiveness of selected stilbene optical brighteners as enhancers of the beet armyworm (lepidoptera: noctuidae) nuclear polyhedrosis virus.the addition of a stilbene optical brightener, tinopal lpw, at 1% concentration (wt:wt) significantly reduced the lc50 of the beet armyworm nuclear polyhedrosis virus (semnpv) from 2.9 pib/mm2 to 0.02 pib/mm2. moreover, the lt50 of semnpv was reduced by 34% by the addition of tinopal lpw. seven other structurally related stilbene brighteners were also tested as viral enhancers. five of these brighteners (tinopal lpw, blankophor bbh, blankophor hrs, blankophor p167, and blankophor rkh) reduced ld ...200111332823
developments in the use of baculoviruses for the surface display of complex eukaryotic proteins.the ability to couple genotype to phenotype has proven to be of immense value in systems such as phage display and has allowed genes encoding novel functions to be selected directly from complex libraries. however, the complexity of many eukaryotic proteins places a severe constraint on successful display in escherichia coli. this restriction could be resolved if a eukaryotic virus could be similarly engineered for display purposes. preliminary data have suggested that the baculovirus autographa ...200111356285
specific binding of baculoviruses displaying gp64 fusion proteins to mammalian cells.viral vectors displaying specific ligand binding moieties have raised an increasing interest in the area of targeted gene therapy. in this report, we describe baculovirus vectors displaying either a functional single chain antibody fragment (scfv) specific for the carcinoembryonic antigen (cea) or the synthetic igg binding domains (zz) derived from protein a of staphylococcus aureus. in addition, the vectors were engineered to incorporate a reporter gene encoding the enhanced green fluorescent p ...200111396970
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