Publications
| Title | Abstract | Year Filter | PMID(sorted ascending) Filter |
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| 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 ... | 1998 | 9811738 |
| 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 ... | 1998 | 9811739 |
| 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 ... | 1998 | 9811740 |
| 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 ... | 1998 | 9811761 |
| 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 ... | 1998 | 9813207 |
| 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 ... | 1998 | 9852042 |
| 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 ... | 1998 | 9856100 |
| [expression of green fluorescent protein with baculovirus vector in insect cells]. | the green fluorescent protein (gfp) gene was subcloned into the transfer vector pvlneo downstream of the polyhedrin gene (ocu) promoter. insect cells were cotransfected with recombinant plasmid and autographa californica nuclear polyhedrosis virus (acnpv) dna. in the presence of g418, the recombinant virus containing gfp gene was purified. the gfp expressed in insect cells with a mw of 30 kda is observable by strong green light under a fluorescent microscope. excitation and emission spectra of t ... | 1997 | 9863195 |
| 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 ... | 1999 | 9878296 |
| 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 ... | 1999 | 9882325 |
| molecular characterization, enzymatic analysis, and purification of murine proprotein convertase-1/3 (pc1/pc3) secreted from recombinant baculovirus-infected insect cells. | a cdna coding for the murine proprotein convertase-1 (mpc1 also known as mpc3 or mspc3) was inserted into the autographa californica nuclear polyhedrosis virus. following infection of spodoptera frugiperda cells, the recombinant n-glycosylated protein is secreted into the cell culture medium from which it can be purified to homogeneity as a fully enzymatically active enzyme. two major secreted molecular forms of mpc1 with apparent molecular weights of 85 and 71 kda, respectively, and a minor one ... | 1998 | 9882569 |
| 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 ... | 1999 | 9887319 |
| an efficient way to introduce unique restriction endonuclease sites into a baculovirus genome. | recombinant baculoviruses which can be linearized at unique sites with restriction endonucleases can greatly facilitate the construction of other recombinants including baculovirus expression vectors and site-specific mutants. we designed a strategy to introduce unique restriction endonuclease sites at virtually any location in a baculovirus genome. the unique sites were first introduced onto a transfer plasmid which also contained in the vector portion of the plasmid an e. coli lacz gene and a ... | 1998 | 9923739 |
| a novel expression system based on host-range expansion of baculovirus. | a host range expanded recombinant autographa californica multiple-nucleocapsid nucleopolyhedrosis virus acmnpv/r2 was obtained by cotransfection of the bacmid dna from escherichia coli dh10bac along with a plasmid pbmh-m containing hindiii m fragment of bombyx mori nuclear polyhedrosis virus (bmnpv) genomic dna. a recombinant transposon vector carrying a mutant green fluorescent protein gene (gfp) and a polyhedrin gene was constructed. transposition was carried out in both e. coli dh10bac and e. ... | 1998 | 9923744 |
| 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 ... | 1999 | 9971831 |
| 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 ... | 1999 | 10074194 |
| optimising fed-batch production of recombinant proteins using the baculovirus expression vector system. | fed-batch culture can offer significant improvement in recombinant protein production compared to batch culture in the baculovirus expression vector system (bevs), as shown by nguyen et al. (1993) and bedard et al. (1994) among others. however, a thorough analysis of fed-batch culture to determine its limits in improving recombinant protein production over batch culture has yet to be performed. in this work, this issue is addressed by the optimisation of single-addition fed-batch culture. this t ... | 1998 | 10099329 |
| 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 ... | 1999 | 10099527 |
| 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 ... | 1999 | 10194406 |
| 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 ... | 1999 | 10201111 |
| 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 ... | 1999 | 10211971 |
| 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 ... | 1999 | 10222179 |
| 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 ... | 1999 | 10233952 |
| coinfection of spodoptera exigua and spodoptera frugiperda cell lines with the nuclear polyhedrosis viruses of autographa californica and spodoptera exigua. | the autographa californica multinucleocapsid nuclear polyhedrosis virus (acmnpv) has a broad host range among lepidoptera. in contrast, the spodoptera exigua mnpv (semnpv) can replicate efficiently only in s. exigua larvae or s. exigua-derived cell lines. in this study, we examined the coinfection of s. exigua se301 and spodoptera frugiperda iplb-sf21aeii (sf21) cell lines with semnpv and acmnpv recombinant (ac360-501beta-gal) which was constructed for expression of beta-galactosidase under cont ... | 1998 | 10325534 |
| 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 ... | 1999 | 10347225 |
| temperature dependent characteristics of a recombinant infectious hematopoietic necrosis virus glycoprotein produced in insect cells. | a recombinant infectious hematopoietic necrosis virus (ihnv) glycoprotein (g protein) was produced in insect cells using a baculovirus vector (autographa californica nuclear polyhedrosis virus). characteristics of this protein were evaluated in relation to native viral g protein. a full-length (1.6 kb) cdna copy of the glycoprotein gene of ihnv was inserted into the baculovirus vector under control of the polyhedrin promoter. high levels of g protein (approximately 0.5 microgram/1 x 10(5) cells) ... | 1999 | 10349547 |
| generation of a new protein purification matrix by loading ceramic hydroxyapatite with metal ions--demonstration with poly-histidine tagged green fluorescent protein. | the gene encoding the green fluorescent protein (gfp) from the jellyfish aequorea victoria, was inserted under transcriptional control of the polyhedrin promoter of the autographa californica nuclear polyhedrosis virus and expressed in the spodoptera frugiperda insect cell line sf9 during viral infection. the baculovirus transfervector pbluebachisb was used for constructing the recombinant baculovirus, so that the green fluorescent protein could be tagged with a poly-histidine tail. this fusion ... | 1999 | 10361722 |
| mitochondrial dna acts as potential promoter of the baculovirus rna polymerase. | we have examined whether mitochondrial dna could act as target of the rna polymerase encoded by the baculovirus autographa californica multicapsid nuclear polyhedrosis virus, because the baculovirus late promoters and the control region of host mitochondrial dna show a high degree of sequence similarity. in vitro transcription using mitochondrial dna from spodoptera frugiperda cells and nuclear extracts prepared from baculovirus infected cells demonstrates that mitochondrial dna is recognized by ... | 1999 | 10384964 |
| 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 ... | 1999 | 10446387 |
| the establishment of new cell lines from pseudaletia unipuncta with differential responses to baculovirus infection. | six insect cell lines from pseudaletia unipuncta embryos were established and characterized, and their susceptibility to autographa californica multiple nuclear polyhedrosis virus (acmnpv) infection was investigated. these embryonic p. unipuncta cell lines had characteristics distinct from each other in morphology and growth, and showed differential responses to acmnpv infection. among the six cell lines, two were highly susceptible to virus infection. one of these two cell lines, bti-pu-a7s, pr ... | 1999 | 10476920 |
| the characterization and phylogenetic relationship of the trichoplusia ni single capsid nuclear polyhedrosis virus polyhedrin gene. | the polyhedrin gene (polh) was identified from the trichoplusia ni (tni) single capsid nuclear polyhedrosis virus (snpv). an ecori fragment containing the truncated polyhedrin gene was detected by hybridization with an acmnpv expression vector probe; the remaining portion of the gene was amplified by reverse pcr. an open reading frame (orf) of 741 nucleotides (nt), encoding a putative protein of 246 amino acids (a.a) with mr 28,780 da was identified. the 5'-noncoding region contained the putativ ... | 1999 | 10499452 |
| [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. | 1999 | 10500990 |
| 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 ... | 1999 | 10510487 |
| detection of baculovirus-infected insect cells by flow cytometric side-scatter analyses. | the baculovirus expression vector system (bevs), utilizing the autographa californica nuclear polyhedrosis virus (acnpv), has turned out to be an attractive alternative for high-level expression (<600 mg/l) of recombinant proteins. however, there is a shortage of reliable methods for monitoring the infection process in situations where marker proteins cannot be used. | 1999 | 10520205 |
| expression of genes coding for animal virus glycoproteins in heterologous systems. | the outermost layers of animal viruses are usually composed of glycoproteins. they are responsible not only for the entrance of viruses into, and release from host cells but also for the initial interaction of a viral particle with immunological defense of the host. it is therefore not surprising that many laboratories devote a lot of effort to study viral glycoproteins at the molecular level. very often such studies are possible only after the introduction of a glycoprotein gene into a heterolo ... | 1999 | 10547034 |
| baculovirus-based genetic screen for antiapoptotic genes identifies a novel iap. | the prototype baculovirus, autographa californica multiple nuclear polyhedrosis virus (acmnpv) expresses p35, a potent anti cell-death gene that promotes the propagation of the virus by blocking host cell apoptosis. infection of insect sf-21 cells with acmnpv lacking p35 induces apoptosis. we have used this pro-apoptotic property of the p35 null virus to screen for genes encoding inhibitors of apoptosis that rescue cells infected with the p35 defective virus. we report here the identification of ... | 1999 | 10593985 |
| diagnosis of penaeus monodon-type baculovirus by pcr and by elisa of occlusion bodies. | the black tiger prawn penaeus monodon is a valuable aquaculture product in taiwan. two specific diagnostic methods were established for p. monodon-type baculovirus, one using polymerase chain reaction (pcr) technology and the other enzyme-linked immunosorbent assay (elisa) technology. monodon-type baculovirus (mbv) was purified by sucrose gradient centrifugation from occlusion bodies of mbv-infected postlarvae of p. monodon. mbv dna was subsequently purified from the occlusion bodies and its pre ... | 2000 | 10782342 |
| differential activity of two non-hr origins during replication of the baculovirus autographa californica nuclear polyhedrosis virus genome. | the identification of potential baculovirus origins of replication (ori) has involved the generation and characterization of defective interfering particles that contain major genomic deletions yet retain their capability to replicate by testing the replication ability of transiently transfected plasmids carrying viral sequences in infected cells. so far, there has not been any evidence to demonstrate the actual utilization of these putative origins in autographa californica multinucleocapsid nu ... | 2000 | 10799593 |
| the autographa californica nuclear polyhedrosis virus p143 gene encodes a dna helicase. | the p143 protein of autographa californica nuclear polyhedrosis virus is essential for replication of viral dna. to determine the function of p143, the protein was purified to near homogeneity from recombinant baculovirus-infected cells that overexpress p143. atpase activity copurified with p143 protein during purification and also during gel filtration at a high salt concentration. the atpase activity did not require the presence of single-stranded dna, but was stimulated fourfold by the additi ... | 2000 | 10799604 |
| the expression and biologic effects of ovine interleukin-4 on t and b cell proliferation. | using the reverse-transcriptase polymerase chain reaction (rt-pcr), cdna encoding ovine (ov) interleukin-4 (ovil-4) was generated from mitogen-stimulated peripheral blood mononuclear cells (pbmc). two identical clones generated from separate rt-pcr reactions differed from a published ovil-4 sequence, although they had a high degree of identity with the bovine and human homologs. we show by sequence analysis that the ovil-4 cdna retained the four alpha-helix structure and disulfide bonds identifi ... | 2000 | 10805377 |
| 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 ... | 2000 | 10856195 |
| expression of highly controllable genes in insect cells using a modified tetracycline-regulated gene expression system. | a modified tetracycline-responsive expression system (tres) for use in insect cells was developed. the tres contains two components: one encodes a tetracycline-controllable transactivator (tta) and the other contains a tet operator dna sequence to drive the luciferase gene. our results show that the human cytomegalovirus (cmv) promoter, an essential part for strong tta expression in mammalian system, was not functional in insect cells. thus further modifications were required. functional tta was ... | 2000 | 10862988 |
| 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 ... | 2000 | 10872882 |
| enhancement in activity of homologous and heterologous baculoviruses infectious to beet armyworm (lepidoptera: noctuidae) by an optical brightener. | the nuclear polyhedrosis virus (npv) from the beet armyworm, spodoptera exigua (hübner) (semnpv), was the most active virus tested against the beet armyworm (lc50 = 4.1 pibs/mm2), followed by nuclear polyhedrosis viruses from the alfalfa looper, autographa californica (speyer) (acmnpv; lc50 = 92.6 pibs/mm2), and the celery looper, anagrapha falcifera (kirby) (afmnpv; lc50 = 195.7 pibs/mm2). in the case of the nuclear polyhedrosis virus from the bollworm, helicoverpa armigera (hübner), lc50s coul ... | 2000 | 10902301 |
| baculoviral display of functional scfv and synthetic igg-binding domains. | viral vectors displaying specific ligand binding moities such as scfv fragments or intact antibodies hold promise for the development of targeted gene therapy vectors. in this report we describe baculoviral vectors displaying either functional scfv fragments or the synthetic z/zz igg binding domain derived from protein a. display on the baculovirus surface was achieved via fusion of the scfv fragment or z/zz domain to the n-terminus of gp64, the major envelope protein of the autographa californi ... | 2000 | 10944446 |
| 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( ... | 2000 | 10960702 |
| presentation of antigenic sites from foot-and-mouth disease virus on the surface of baculovirus and in the membrane of infected cells. | we describe the construction of recombinant baculoviruses displaying on their surface and in the membrane of infected cells the small, immunodominant antigenic site (site a) or the large polyprotein (p1) coding for the four structural proteins of foot-and-mouth disease virus (fmdv). the coding sequences were inserted in the amino-terminus of gp64, the major glycoprotein of the baculovirus autographa californica nuclear polyhedrosis virus (acnpv). following infection of insect cells with the reco ... | 2000 | 11043943 |
| 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 ... | 2000 | 11117998 |
| 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 ... | 2000 | 11142292 |
| production of biologically active recombinant bovine interferon-gamma by two different baculovirus gene expression systems using insect cells and silkworm larvae. | the full-length bovine interferon-gamma (bifn-gamma) cdna, including the secretion signal peptide coding region was recloned into baculovirus transfer vectors pacym1 and pbm050. these vectors were co-transfected with autographa californica nuclear polyhedrosis virus (acnpv) or bombyx mori nuclear polyhedrosis virus (bmnpv) dna into spodoptera frugiperda cells (sf21ae) and bombyx mori cells (bmn), respectively. the recombinant viruses, named acbifn-gamma and bmbifn-gamma, were then recovered. rec ... | 2001 | 11145838 |
| comparative effects of a genetically engineered insect virus and a growth-regulating insecticide on microbial communities in aquatic microcosms. | the effects of a genetically engineered insect baculovirus on indigenous aquatic microbial communities were determined in closed, recirculating aquatic microcosms, and compared with the effects of a natural strain of the virus and of a growth-regulating insecticide, dimilin. the recombinant virus was a nuclear polyhedrosis virus (npv) of the spruce budworm (choristoneura fumiferana (cf)) with a lacz marker gene inserted into the egt region of the cfnpv. the natural virus was ireland strain cfnpv ... | 2001 | 11161682 |
| high-level expression of human acidic fibroblast growth factor and basic fibroblast growth factor in silkworm (bombyx mori l.) using recombinant baculovirus. | a hybrid of autographa californica nuclear polyhedrosis virus and bombyx mori nuclear polyhedrosis virus, which is infectious to both spodoptera frugiperda and bombyx mori, was prepared in our previous study. two recombinant hybrid baculoviruses, carrying cdnas of human acidic and basic fibroblast growth factors, respectively, were successfully constructed in this study, for the large-scale production of human afgf and bfgf using silkworm as host. these recombinant viruses were used to inoculate ... | 2001 | 11162406 |
| diagnostic immunoassays for tick-borne encephalitis virus based on recombinant baculovirus protein expression. | the baculovirus expression system that utilizes autographa californica nuclear polyhedrosis virus was used to express the highly antigenic envelope protein e of a tick-borne encephalitis (tbe) complex virus, as well as a c-terminally truncated form of protein e (etr). the recombinant proteins were produced with a histidine-tag at their carboxy-terminus. protein purification by nickel agarose chromatography resulted in high concentrations of pure etr protein, but only poor yields of e protein. th ... | 2001 | 11164488 |
| [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 ... | 2000 | 11209691 |
| 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 ... | 2001 | 11233097 |
| dynamic nuclear localization of the baculovirus proteins ie2 and pe38 during the infection cycle: the promyelocytic leukemia protein colocalizes with ie2. | the early gene products ie2 and pe38 of autographa californica multicapsid nuclear polyhedrosis virus localize to distinct nuclear domains after transient expression. here, the nuclear localization pattern and the putative association with cellular proteins have been determined during virus infection to shed light on the functional significance of the nuclear domains. ie2 was always localized to distinct nuclear structures while pe38 was partly present in nuclear dots. confocal imaging indicated ... | 2001 | 11262179 |
| actin rearrangement-inducing factor of baculoviruses is tyrosine phosphorylated and colocalizes to f-actin at the plasma membrane. | in previous studies we have identified actin rearrangement-inducing factor 1 as an early gene product of autographa californica multicapsid nuclear polyhedrosis virus that is involved in the remodeling of the actin cytoskeleton. we have constructed viral recombinants with a mutated arif-1 open reading frame that confirm the causal link of arif-1 expression and the actin rearrangement observed as accumulation of f-actin at the plasma membrane at 3 to 7 h postinfection. infection with arif mutant ... | 2001 | 11264366 |
| novel sp family-like transcription factors are present in adult insect cells and are involved in transcription from the polyhedrin gene initiator promoter. | we earlier documented the involvement of a cellular factor, polyhedrin (polh) promoter-binding protein, in transcription from the autographa californica nuclear polyhedrosis virus polh gene promoter. sequences upstream of the polh promoter were found to influence polh promoter-driven transcription. analysis of one such region, which could partially compensate for the mutated polh promoter and also activate transcription from the wild-type promoter, revealed a sequence (acsp) containing a caccc m ... | 2001 | 11294840 |
| combination treatment for osteosarcoma with baculoviral vector mediated gene therapy (p53) and chemotherapy (adriamycin). | the insect baculovirus autographa californica multiple nuclear polyhedrosis virus (acmnpv) has been evaluated as a vector for gene delivery to human tumor cells. a human osteogenic sarcoma cell line, saos-2, was found to be highly susceptible to infection with a baculoviral vector, with nearly 100% of saos-2 cells being able to express a lacz reporter gene after a brief exposure to the virus at a m.o.i. of 30 pfu/cell. the production of beta-galactosidase protein was 18-times greater than that i ... | 2001 | 11322486 |
| 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 ... | 2001 | 11332823 |
| developments in the use of baculoviruses for the surface display of complex eukaryotic proteins. | the ability to couple genotype to phenotype has proven to be of immense value in systems such as phage display and has allowed genes encoding novel functions to be selected directly from complex libraries. however, the complexity of many eukaryotic proteins places a severe constraint on successful display in escherichia coli. this restriction could be resolved if a eukaryotic virus could be similarly engineered for display purposes. preliminary data have suggested that the baculovirus autographa ... | 2001 | 11356285 |
| 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 ... | 2000 | 11395967 |
| specific binding of baculoviruses displaying gp64 fusion proteins to mammalian cells. | viral vectors displaying specific ligand binding moieties have raised an increasing interest in the area of targeted gene therapy. in this report, we describe baculovirus vectors displaying either a functional single chain antibody fragment (scfv) specific for the carcinoembryonic antigen (cea) or the synthetic igg binding domains (zz) derived from protein a of staphylococcus aureus. in addition, the vectors were engineered to incorporate a reporter gene encoding the enhanced green fluorescent p ... | 2001 | 11396970 |
| a new continuous cell line from larval hemocytes of spodoptera litura (f.). | a new cell line has been established from larval hemocytes of the moth, s. litura (tobacco cut worm). it took 147 days to form a monolayer and one year for the first 17 passages. at present, the culture is at 86th passage level and is designated niv-su-1095. three cell types could be distinguished, viz. plasmatocytes (53%), prohemocytes (36%) and granular hemocytes (11%). the chromosome number was very high, 74% metaphase cells showed more than 100 chromosomes. the cells could be cryopreserved. ... | 2000 | 11411040 |
| expression and one-step purification of intracellular human prolactin in insect cells. | human prolactin was expressed in insect culture cells by recombinant baculoviruses carrying prolactin gene cdna placed under the transcriptional control of polyhedrin gene promoter of autographa californica nuclear polyhedrosis virus. preliminary results of recombinant human prolactin expression as extracellular as well as intracellular product of baculovirus expression system were presented at the febs meeting in nice, france, in 1999 (abstracts, p. 288). in the present work prolactin was expre ... | 2001 | 11437600 |
| [a 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 ... | 2000 | 11460790 |
| 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 ... | 2001 | 11507209 |
| identification, sequence analysis, and phylogeny of the immediate early gene 1 of the trichoplusia ni single nucleocapsid polyhedrosis virus. | substantial research has been conducted on the immediate early i (ie-1) genes from the prototype baculovirus auographa californica multicapsid nuclear polyhedrosis virus (acmnpv) and the orgyia pseudotsugata multicapsid nuclear polyhedrosis virus (opmnpv). in both cases ie-1 gene products have been implicated in transcriptional activation and repression. in this study an ie-1 homolog was identified from trichoplusia ni single nucleocapsid polyhedrosis virus (tnisnpv). nucleotide sequence analysi ... | 2001 | 11556402 |
| investigation of sequential behavior of carboxyl protease and cysteine protease activities in virus-infected sf-9 insect cell culture by inhibition assay. | proteases produced during the culture of spodoptera frugiperda sf-9 cells infected with autographa californica nuclear polyhedrosis virus (acnpv) were assayed with various protease inhibitors. this inhibitory analysis revealed that: (1) carboxyl and cysteine proteases were predominantly produced by the insect cells infected with recombinant acnpv, the gene of which encoded a variant of green fluorescent protein in a portion of the polyhedrin gene of the baculovirus, and (2) the protease activity ... | 2001 | 11601624 |
| effects of deletion and overexpression of the autographa californica nuclear polyhedrosis virus fp25k gene on synthesis of two occlusion-derived virus envelope proteins and their transport into virus-induced intranuclear membranes. | partial deletions within autographa californica open reading frame 61 (fp25k) alter the expression and accumulation profile of several viral proteins and the transport of occlusion-derived virus (odv)-e66 to intranuclear membranes during infection (s. c. braunagel et al., j. virol. 73:8559-8570, 1999). here we show the effects of a full deletion and overexpression of fp25k on the transport and expression of two odv envelope proteins, odv-e66 (e66) and odv-e25 (e25). deletion and overexpression o ... | 2001 | 11602724 |
| persistence of an occlusion-negative recombinant nucleopolyhedrovirus in trichoplusia ni indicates high multiplicity of cellular infection. | we use data from the serial passage of co-occluded recombinant autographa californica nuclear polyhedrosis virus (acmnpv) to estimate the viral multiplicity of infection of cells within infected insects. co-occlusion, the incorporation of wild-type and mutant virus genomes in the same occlusion body, has been proposed as a strategy to deliver genetically modified viruses as insecticides in a way that contains their spread in the environment. it may also serve as a means whereby naturally occurri ... | 2001 | 11679346 |
| identification of apoptosis-inhibiting gene in leucania separata nuclear polyhedrosis virus. | a novel gene lsp40 from leucania separata nuclear polyhedrosis virus (lsnpv) which was homologous to the p35 gene from autographa californica nuclear polyhedrosis virus (acnpv) was localized in the ecorv-5.5 kb fragment of lsnpv genome dna and was sequenced. the open reading frame (orf) of lsp40 was 906 bp long and encoded an approximately 40 kda peptide consisting of 302 amino acid residues. the isp40 shares 80.4% and 70.4% identity of nucleotide and amino acid sequence, respectively, to the ac ... | 2001 | 11765917 |
| [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 ... | 2000 | 11767686 |
| inhibitor of apoptosis protein from orgyia pseudotsugata nuclear polyhedrosis virus provides a costimulatory signal required for optimal proliferation of developing thymocytes. | the inhibitors of apoptosis proteins (iaps) constitute a family of endogenous inhibitors that control apoptosis in the cell by inhibiting caspase processing and activity. iaps are also implicated in cell division, cell cycle regulation, and cancer. to address the role of iaps in thymus development and homeostasis, we generated transgenic mice expressing iap generated from the baculovirus orgyia pseudotsugata nuclear polyhedrosis virus (opiap). developing thymocytes expressing opiap show increase ... | 2002 | 11823509 |
| biosynthesis and localization of the autographa californica nuclear polyhedrosis virus 25k gene product. | mutations of the acmnpv 25k gene are associated with the "few polyhedra" phenotype (m. j. fraser et al., 1983, j. virol. 47, 287-300; b. beames and m. d. summers, 1989, virology 168, 344-353). polyclonal antisera was produced and used to investigate the time course of expression and localization of the 25k protein in infected cells. western blot analysis detected 25k protein in both cytosolic and nuclear extracts from 18-24 hr p.i. through 96 hr p.i. and also in purified viral occlusions, but no ... | 1995 | 11831709 |
| passage of autographa californica nuclear polyhedrosis virus through the midgut epithelium of spodoptera exigua larvae. | a special recombinant of autographa californica multicapsid nuclear polyhedrosis virus (acnpv) was designed to study the early histopathological events of baculovirus infection in spodoptera exigua larvae. this recombinant contained a drosophila melanogaster heat shock 70 promoter driving an escherichia coli beta-galactosidase (lac-z) reporter gene to monitor the presence of early viral gene expression and a second reporter gene, the e. coli beta-glucuronidase (gus) gene, under control of the ve ... | 1995 | 11831715 |
| rat monoclonal antibodies differentiating between the epstein-barr virus nuclear antigens 2a (ebna2a) and 2b (ebna2b). | rat monoclonal antibodies were produced against the c-terminus of epstein-barr virus nuclear antigens 2a (ebna2a) and 2b (ebna2b) expressed as bacterial trpe fusion proteins. the initial screening was performed using a soluble bacterial extract containing the fusion proteins. positive hybridomas were confirmed by immunofluorescence on sf158 (spodoptera frugiperda) insect cells infected with recombinant baculovirus (autographa californica nuclear polyhedrosis virus) and expressing the complete eb ... | 1995 | 11831716 |
| intron-dependent stimulation of marker gene expression in cultured insect cells. | we tested in a systematic fashion the effect of an intron on the level of luciferase expression in cultured c6/36 aedes albopictus cells. the intron was inserted in both orientations, upstream and downstream of the luciferase coding region in two different luciferase expression vectors. the two parental luciferase expression vectors differed only in their promoters, one containing the drosophila melanogaster actin5c promoter and the other the autographa californica nuclear polyhedrosis virus hr5 ... | 2002 | 11841506 |
| characterization of a truncated soluble form of the baculovirus (acmnpv) major envelope protein gp64. | a truncated tagged form of the autographica californica multiple nuclear polyhedrosis virus major surface glycoprotein, gp64, has been expressed using the baculovirus expression system and purified to homogeneity by immune-affinity chromatography. the protein, which is responsible for virus-cell fusion, was a trimer in solution and retained this oligomeric form at ph 5, the ph of fusion. circular dichroism spectroscopy indicated a protein with mixed alpha-helix and beta-sheet content that did no ... | 2002 | 11858713 |
| nuclear ie2 structures are related to viral dna replication sites during baculovirus infection. | the ie2 gene of autographa californica multicapsid nuclear polyhedrosis virus is 1 of the 10 baculovirus genes that have been identified as factors involved in viral dna replication. ie2 is detectable in the nucleus as one of the major early-expressed proteins and exhibits a dynamic localization pattern during the infection cycle (d. murges, i. quadt, j. schröer, and d. knebel-mörsdorf, exp. cell res. 264:219-232, 2001). here, we investigated whether ie2 localized to regions of viral dna replica ... | 2002 | 11967334 |
| in vivo expression and immunological studies of the 42-kilodalton carboxyl-terminal processing fragment of plasmodium falciparum merozoite surface protein 1 in the baculovirus-silkworm system. | the 42-kda carboxyl-terminal processing fragment of plasmodium falciparum merozoite surface protein 1 (msp-1(42)) is an anti-erythrocytic stage malaria vaccine candidate. in this study, msp-1(42) was expressed by using the bombyx mori nuclear polyhedrosis virus-silkworm expression system, and the antigenicity and immmunogenicity of the recombinant protein, bmp42, were evaluated. the average yield of bmp42, as determined by a sandwich enzyme-linked immunosorbent assay (elisa), was 379 microg/ml o ... | 2002 | 12010962 |
| [biosynthesis of a single peptide chain containing human chorionic gonadotropin beta and ovine common alpha subunits tandem]. | hcg beta-olh alpha chimeric cdna was constructed by using overlapping pcr to contact the codons of c-terminal end of hcg beta with the codons of n-terminal end of olh alpha, then it was subcloned into nuclear polyhedrosis virus (acnpv) expression vector pvl1393 to construct expression vector pvl1393-hcg beta-olh alpha. the insect cells (sf9) were cotransfected by the expression vector pvl1393-hcg beta-olh alpha and baculogold acnpv linearized genomic dna, and recombinant viruses acnpv-hcg beta-o ... | 1998 | 12014113 |
| 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 ... | 2001 | 12040393 |
| 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 ... | 2000 | 12058207 |
| development of a competitive enzyme-linked immunosorbent assay for detection of turkey coronavirus antibodies. | a competitive enzyme-linked immunosorbent assay (celisa) was developed for detection of turkey coronavirus (tcv) antibodies. the celisa utilized a recombinant baculovirus (autographa californica nuclear polyhedrosis virus)-expressed tcv nucleocapsid (n) protein and biotin-labeled tcv n protein-specific monoclonal antibody. sensitivity and specificity of the celisa for detection of tcv antibodies were determined by comparison with the indirect fluorescent antibody test (ifat) with 1269 reference, ... | 2002 | 12061642 |
| timing of transmission and the evolution of virulence of an insect virus. | we used the nuclear polyhedrosis virus of the gypsy moth, lymantria dispar, to investigate whether the timing of transmission influences the evolution of virulence. in theory, early transmission should favour rapid replication and increase virulence, while late transmission should favour slower replication and reduce virulence. we tested this prediction by subjecting one set of 10 virus lineages to early transmission (early viruses) and another set to late transmission (late viruses). each linea ... | 2002 | 12061960 |
| baculovirus as mammalian cell expression vector for gene therapy: an emerging strategy. | the monopoly of insect cells to host baculovirus autographa californica multiple nuclear polyhedrosis virus (acmnpv) as a eukaryotic gene expression system has been shattered with the growing evidence that it also infects mammalian cells in culture. although acmnpv fails to replicate in vertebrate cells, it does express alien genes with levels of expression that are dependent on the strength of the promoter used to drive transcription of the foreign gene. it also has been reported that the recom ... | 2002 | 12095297 |
| 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, ... | 2000 | 12098800 |
| 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 ... | 1999 | 12110934 |
| 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. | 1999 | 12114994 |
| 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 ... | 1999 | 12142920 |
| nucleotide sequence and characterization of the p10 gene of spodoptera litura nuclear polyhedrosis virus. | the p10 gene was localized within the 3.0 kbp bamh i fragment of the sinpv genome. with a coding sequence of 318 nucleotides, corresponding to a protein of 105 amino acids, the p10 gene of sinpv is the longest p10 gene identified so far in npvs. the sinpv p10 gene is also distinct in having two a/tttgta motifs in the promoter region. with 10 heptad repeats, the sinpv p10 protein is probably forming a relatively large coiled-coil structure. the organization of sinpv orf552-p10 gene-orf945 cluster ... | 1998 | 12167986 |
| expression of human lactoferrin cdna in insect cells. | a transfer plasmid p8hlfc harboring human lactoferrin cdna was constructed and co-transfected with autographa californica nuclear polyhedrosis virus ( acnpv bacpak6 ) dna into spodoptera frugiperda (sf) cells. the recombinant virus acnpv-hlfc was identified and purified three times by dot hybridization and plaques screening. the expression of human lactoferrin cdna in insect cells was detected by western blotting. the expression level of lactoferrin is about 2% of insect cellular total soluble p ... | 1998 | 12167991 |
| cloning of etp28 gene of eimeria tenella (guangdong strain) sporozoites and its expression in baculovirus expression system. | a refractile body antigen etp28 gene of eimeria tenella (guangdong strain) sporozoites was cloned by pcr from the synthesized first strand cdna. it has a high homology with the previously reported etp28 gene of merck strain ls18. the gene was expressed through standard procedures in the modified autographa californica nuclear polyhedrosis virus (acmnpv-occ(-)) expression system and large amount of heterologous fusion protein (gst-6xhis-etp28) was obtained. the expression product was about 21.3% ... | 1998 | 12174264 |
| nucleotide sequence analysis of the hcnpv cysteine protease gene. | the cysteine protease (cp) gene of hyphantria cunea nuclear polyhedrosis virus (hcnpv) has been located at the hind iii--3.5 kb fragment. the nucleotide sequence has been determined. the results indicated that its open reading frame of 975 nt encoded the protein of 324 amino acids. the baculovirus late gene promoter taag motif was identified at -24 nt upstream from the start codon atg; the typical poly (a) signal sequence aataaa was detected at 33 nt downstream from the stop codon taa. the hcnpv ... | 1998 | 12174267 |
| [identification of functional region of helicase gene promoter in bombyx mori nuclear polyhedrosis virus]. | dna helicases are essential for replication of baculoviruses. it was found that the helicase gene promoter of bombyx mori nuclear polyhedrosis virus, including 510 bp upstream of atg, had both early and late rna initiation sites and could be recognized by cellular rna polymerase. transient expression assays in uninfected sf-21 cells indicated that the helicase gene promoter could be classified as a delayed-early gene promoter. deletion analysis by pcr showed that the regulation region of its bas ... | 2002 | 12198556 |
| production of recombinant ovine interferon tau using a bombyx mori nuclear polyhedrosis baculovirus expression system. | ovine interferon tau (oifn-tau) is an embryonic protein of critical importance in the establishment of pregnancy in the sheep. we have produced recombinant (r) oifn-tau using a baculovirus expression system and demonstrated the biological activity of the protein produced. bombyx mori larvae were infected with b. mori nuclear polyhedrosis virus (bmnpv), modified by inserting a cdna coding for oifn-tau downstream of the strong polyhedron promoter. following infection, antiviral activity of the hae ... | 2002 | 12212938 |
| nucleotide sequence analysis of hasnpv protein kinase. | the protein kinase gene of helicoverpa armigera single nucleocapsid nuclear polyhedrosis virus (hasnpv) has been cloned and sequenced. it is located approximately 1.25 kb downstream of the polyhedrin gene. the predicted molecular mass of this 267 amino acid protein (havpk) is 31 kd. havpk shows a 43.0% amino acid homology to vpk from lymantria dispar multinucleocapsid nuclear polyhedrosis virus (ldmnpv) and a 39.0% homology to pk-1 from autographa californica multinucleocapsid nuclear polyhedros ... | 1997 | 12219213 |
| studies on the expression of the 28 kd glutathione s-transferase gene from schistosoma japonicum in the silkworm (bombyx mori) larvae and insect cells. | the gene of the 28 kd glutathione s-transferase (gst) from the chinese strain of schistosoma japonicum had been expressed in the silkworm (bombyx mori) cells and larvae by bombyx mori nuclear polyhedrosis virus (bmnpv) vector which had been modified. the gst gene was inserted into the right position of the bmnpv genome as identified by southern hybridization. the product was a 28 kd protein, had gst activity and antigenicity. the yield in bmn cells (1x10(6) cells/ml) was 0.77 mg and more than 5 ... | 1997 | 12219234 |
| foreign insect hormones stimulating the transcription of the ie-1 promoter of bombyx mori nuclear polyhedrosis virus in vivo and in vitro. | via a transient expression assay system, an experimental study was undertaken to characterize the effects of insect ecdysone and juvenile hormone analogue on the transient expression of the luciferase gene under the control of the immediate-early gene (ie-1) promoter of bombyx mori nuclear polyhedrosis virus. the results demonstrated that the transcriptional activity of the ie-1 promoter was increased to a certain extent by different insect hormone treatments in uninfected insect cells or fifth ... | 2002 | 12224632 |