Publications

TitleAbstractYear(sorted descending)
Filter
PMID
Filter
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 ...199511831716
characterization of the baculovirus choristoneura fumiferana multicapsid nuclear polyhedrosis virus p10 gene indicates that the polypeptide contains a coiled-coil domain.the dna sequence and transcription pattern of the p10 gene from the spruce budworm baculovirus choristoneura fumiferana multicapsid nuclear polyhedrosis virus (cfmnpv) were analysed. the cfmnpv p10 gene codes for a protein 81 amino acids in length: this is the shortest p10 peptide identified thus far. a novel characteristic of the cfmnpv p10 gene is that it contains tandem late initiation sites (taag) having different temporal transcription patterns. comparative analysis of cfmnpv p10 and the am ...19958847496
identification and characterization of an early gene in the lymantria dispar multinucleocapsid nuclear polyhedrosis virus.the lymantria dispar multinucleocapsid nuclear polyhedrosis virus (ldmnpv) gene encoding g22 was cloned and sequenced. the g22 gene codes for a 191 amino acid protein with a predicted m(r) of 22,000. expression of g22 in a rabbit reticulocyte system generated a protein with an m(r) of 24,000, in close agreement with the molecular mass predicted from the nucleotide sequence. g22 is not significantly homologous to any known protein, nor is a g22 homologue present in the autographa californica mnpv ...19958847497
in vivo and in vitro characterization of several isolates of spodoptera exigua nuclear polyhedrosis virus.spodoptera exigua nuclear polyhedrosis viruses (senpvs), isolated form five geographically distinct regions of japan and thailand, were characterized by their dna restriction endonuclease pattern, level of virus production in a continuous cell line of s. exigua and biological activity to s. exigua larvae. the ecori and psti fragments exhibited similar overall patterns with minor differences. digestion of virus dna from a plaque-purified isolate, senpv-i1, with psti yielded 14 fragments and the e ...19958825303
establishment of a cell line derived from embryos of the potato tuber moth phthorimaea operculella (zeller).a cell line from the main insect pest of potatoes in tropical and subtropical areas, phthorimaea operculella (zeller), was obtained from embryoculture. these cells were cultured in grace's modified medium. the cell line, designated ors-pop-93, had a heterogeneous population consisting of spherical and spindle cells with great capacity to adhere and a doubling time of 40 h. they were subcultured for more than 60 passages. their polypeptidic profile was different from profiles of other lepidoptera ...19958826086
identification and nucleotide sequence of an ecdysteroid udp-glucosyltransferase gene of spodoptera littoralis multicapsid nuclear polyhedrosis virus.the spodoptera littoralis multicapsid nuclear polyhedrosis virus (slmnpv) is a member of the baculoviridae that shows a distant genetic relationship to the prototype autographa californica mnpv (acmnpv). using an acmnpv gene-specific probe, we identified and mapped an ecdysteroid udp-glucosyltransferase (egt) gene in the genome of slmnpv. sequence determination of a part from the hybridizing dna fragment revealed an open reading frame of 1548 nucleotides that exhibits 38% and 44% identity to the ...19958808334
high expression of hbv s gene in bombyx mori cell culture and in silkworms.the coding sequence of human hepatitis b virus (hbv) subtype adr surface antigen s gene was inserted into the genome of the nuclear polyhedrosis virus of bombyx mori (bmnpv) under control of the polyhedrin promoter, and a recombinant bmnpvs virus was constructed. upon infection of the recombinant virus, the yield of hbsag was 35.5 micrograms per 10(6) cells, 750 micrograms per larva, and 690 micrograms per pupa. western blot and electron microscope observation of the preliminary purified hbsag p ...19958679930
[sequencing of polyhedrin gene of leucania separata multiple capsid nuclear polyhedrosis virus by silver sequencing system].the genome of lsnpv was digested with ecorv and cloned into pbluescript. with the partial polyhedrin gene of autographa californica as a probe, positive recombinants were screened by colony in situ hybridization, further characterized by sequencing, and designated as plsev and plsph5. the intact 741 bp polyhedrin gene of lsnpv is located on the two recombinants revealed by silver sequencing system, and shares 80.0% and 97.0% nucleotide identity of the coding sequence, 87.8% and 97.5% amino acid ...19958701583
expression and radiolabeling of human c-reactive protein in baculovirus-infected cell lines and trichoplusia ni larvae.human c-reactive protein (crp) is a member of the pentraxin family of proteins which are molecules composed of five identical subunits arranged in a planar configuration. in the present study a human crp cdna clone was ligated into the baculovirus vector pvl1393 which was used to establish a recombinant strain of baculogold autographa californica multiple nuclear polyhedrosis virus containing the coding and leader sequence for human crp (designated acmnpv-crp). synthesis and secretion of crp wer ...19958527929
construction and insect larval expression of recombinant vesicular stomatitis nucleocapsid protein and its use in competitive elisa.the gene encoding the nucleocapsid (n) protein of indiana 1 serotype vesicular stomatitis virus (vsv-in1) was transferred into the genome of autographa californica nuclear polyhedrosis virus (baculovirus) as a full-length non-fusion construct under the control of the polyhedrin gene promoter. recombinant n protein was obtained from trichoplusia ni insect larvae inoculated 72-96 h previously with the recombinant baculovirus. polyclonal antibody (pab) against vsv-in1 was produced in mice using vsv ...19958530566
convenient desktop-scale production of the extracellular domain of the human growth hormone receptor by an insect-baculovirus secretion system using a protein-free culture.expression of a gene encoding the extracellular domain of the human growth hormone receptor (hghr-ed) inserted into the genome of autographa californica nuclear polyhedrosis virus was done using a desktop-scale spinner culture. spodoptera frugiperda 9 (sf9) cells infected with the recombinant virus secreted a protein with hgh-binding activity into the medium. oxygen supplementation was required for high level secretion of the product. the highest cell production capability was estimated at more ...19958534995
identification and characterization of the v-cath gene of the baculovirus, cfmnpv.the v-cath gene of the autographa californica multi-nucleocapsid nuclear polyhedrosis virus (acmnpv) encodes a cathepsin l-like proteinase which plays a role in the liquefaction of host tissues during a viral infection [1]. we have identified a homologous gene in the spruce budworm virus, choristoneura fumiferana mnpv (cfmnpv). the cfmnpv v-cath gene is 74% identical to acmnpv v-cath at the nucleotide sequence level and 80% identical at the level of predicted amino acid sequence. transcription a ...19958547313
high-level expression of human beta-interferon gene in the silkworm with new constructed bmnpv vector.bombyx mori nuclear polyhedrosis virus (bmnpv) and bombyx mori cells as well as silkworm larvae were used successfully for the production of biologically active recombinant proteins. there are only a few types of bmnpv general vectors. here a new type of vector plasmid pbm92 was constructed in this experiment. the translational initiation codon atg of the polyhedrin gene in pbm92 was changed into att, and then five cloning sites of a foreign gene were ligated after the +12 bp site of the polyhed ...19958562846
production of polyhedra of the autographa californica nuclear polyhedrosis virus using the sf21 and tn5b1-4 cell lines and comparison with host-derived polyhedra by bioassay.both wild-type and recombinant baculoviruses are becoming more attractive for the control of insect pests. thus, there is an increased incentive to address and resolve logistical problems associated with large-scale production of these viruses. in this study, we have compared the potential of two insect cell lines, tn5b1-4 and sf21, for the production of polyhedra and compared the efficacy of both cell culture-derived and host-derived viruses by bioassay. the efficacy of both wild-type acmnpv an ...19958568278
nucleotide sequence of a 1446 base pair sali fragment and structure of a novel early gene of leucania separata nuclear polyhedrosis virus.a 1446 bp sali fragment of lsnpv was sequenced by the silver staining method, and two large open reading frames (orfs, orf1 and orf2) were found, both contain typical characteristics of the 5' regulatory elements of baculovirus early genes. orf1 is 345 bp long with the capacity to encode a putative protein of 114 amino acid residues with mw about 13 kda and was designated p13 gene, orf2 comprises 248 bp from the 3' end of the fragment. in the untranslated region (utr) of orf1, a 33 bp mini cistr ...19958572949
nucleocapsid protein n of lelystad virus: expression by recombinant baculovirus, immunological properties, and suitability for detection of serum antibodies.the orf7 gene, encoding the nucleocapsid protein n of lelystad virus (lv), was inserted downstream of the p10 promoter into autographa californica nuclear polyhedrosis virus (baculovirus). the resulting recombinant baculovirus, designated bac-orf7, expressed a 15-kda protein in insect cells. this protein was similar in size to the n protein expressed by lv in cl2621 cells when it was analyzed on sodium dodecyl sulfate-polyacrylamide gels. the n protein expressed by bac-orf7 was immunoprecipitate ...19958574824
two human alpha 2-adrenoceptor subtypes alpha 2a-c10 and alpha 2b-c2 expressed in sf9 cells couple to transduction pathway resulting in opposite effects on camp production.the baculovirus expression vector system utilizing the strong polyhedrin gene promoter of the autographa californica nuclear polyhedrosis virus (acnpv) was used for high level expression of the two alpha 2-adrenoceptor subtypes alpha 2a-c10 and alpha 2b-c2 in spodoptera frugiperda (sf-9) insect cells. for rapid screening of recombinant viruses the luciferase gene was expressed under the early etl-promoter (early transcript large) in the same plasmid. both receptor subtypes showed the same rank o ...19958575536
replication of a mamestra brassicae nuclear polyhedrosis virus in a newly established mamestra brassicae cell line.a continuous cell line, designated mbl-3, was newly established from minced neonate larvae of the cabbage armyworm, mamestra brassicae. this new cell line is heteroploid, containing triploid cells predominantly at the frequency of about 50%. doubling time of the cell population is 33 hrs. the akutsu isolate of a mamestra brassicae nuclear polyhedrosis virus (mbnpv) was examined for replication in 16 continuous cell lines, including the cell line mbl-3, from seven lepidoptera species: mamestra br ...19958578995
expression of human stem cell factor in the baculovirus expression system.a partial cdna of human stem cell factor (scf) was isolated and transferred into the genome of bombyx mori nuclear polyhedrosis virus adjacent to the polyhedrin promoter. the soluble form of the scf protein was expressed in bombyx mori cells and silkworm larvae and was secreted into the medium in an active state. the recombinant scf was similar to natural scf with respect to its interaction with monoclonal antibody and its synergistic biological activity in conjunction with colony-stimulating fa ...19958589646
the baculovirus cysteine protease has a cathepsin b-like s2-subsite specificity.autographa californica nuclear polyhedrosis virus (acnpv) encodes a functional cysteine protease of the papain family which is expressed after infection in spodoptera fruglperda sf9 cells. the protease displays an inhibition profile typical for cysteine proteases and is highly active against synthetic peptide substrates. the ph optimum of the bell-shaped ph-activity curve is between 5.0 and 5.5. the best substrate tested is z-arg-arg-mca which is specific for cathepsin b. the specificity constan ...19958590630
differential transcription of baculovirus late and very late promoters: fractionation of nuclear extracts by phosphocellulose chromatography.an in vitro transcription system based on cytidine-free cassette was developed for the late 39k gene and the very late polyhedrin gene of autographa californica nuclear polyhedrosis virus (acnpv). optimization of transcription conditions revealed that a preincubation step was not required for transcription of late and very late promoters, although preincubation was essential for efficient transcription from an early promoter. the 39k and polyhedrin constructs were actively transcribed by nuclear ...19957707516
yeast-prepro-alpha-factor-leader-region-directed synthesis and secretion of truncated human macrophage colony-stimulating factor in the silkworm bombyx mori.human macrophage colony-stimulating factor (hm-csf) cdna joined to the leader region of the precursor of the yeast mating pheromone alpha-factor (mf alpha l) was expressed at high levels in bmn cells and in silkworm (bombyx mori) larvae, using recombinant bombyx mori nuclear polyhedrosis virus, as a vector. the biological activity of rhm-csf detected in the haemolymph was 1 x 10(6) colony-formation units/ml, approximately half of the expression level directed by the native signal peptide of hm-c ...19957710703
overlapping tata-dependent and tata-independent early promoter activities in the baculovirus gp64 envelope fusion protein gene.in previous studies to characterize basal and activated transcription from the early promoter of the gp64 envelope fusion protein (efp) gene of the orgyia pseudotsugata multicapsid nuclear polyhedrosis virus, the tata box was identified as a functional element, essential for basal transcription from a minimal promoter construct. in the current study, we used discrete deletions and multiple point mutations that removed the functional tata box from larger promoter constructs of the gp64 efp gene t ...19957853477
early transcription of the ie-1 transregulator gene of autographa californica nuclear polyhedrosis virus is regulated by dna sequences within its 5' noncoding leader region.the ie-1 gene of autographa californica nuclear polyhedrosis virus (acmnpv) encodes a transregulatory protein (ie1) which accelerates the expression of early and late virus genes. transcription of ie-1 occurs immediately upon infection from a conserved cagt motif and continues into the late phases. to examine the mechanisms by which ie-1 expression is regulated, cis-acting control elements within the ie-1 promoter were identified by constructing hybrid early promoters and by using site-directed ...19957983706
lymantria dispar nuclear polyhedrosis virus homologous regions: characterization of their ability to function as replication origins.homologous regions (hrs) were identified in the lymantria dispar nuclear polyhedrosis virus (ldmnpv) genome. a 1.58-kb region surrounding hr4 was sequenced and found to have two distinct domains. domain i (about 600 bp) is composed of seven repeats of about 80 bp including a series of palindromes containing mlui sites and overlapping xhoi and saci sites. domain ii (about 700 bp) is composed of eight partially repeated sequences of 60 to 100 bp containing a 15- to 25-bp sequence that is 80 to 100 ...19957983712
codon usage in the prototype baculovirus--autographa californica nuclear polyhedrosis virus.the overall codon usage profile of autographa californica nuclear polyhedrosis virus (acnpv) was analyzed, using uwgcg sequence analysis software package from the known protein coding gene sequences available in genbank release 72. the analysis revealed that although only 45% of the codon used by acnpv have g/c at wobble base position, 15 out of 20 acnpv codons over-utilized for their given amino acids has g/c at the wobble position indicating a possible selection of these codons. the difference ...19958714213
maturation of the dengue-2 virus ns1 protein in insect cells: effects of downstream ns2a sequences on baculovirus-expressed gene constructs.a series of recombinant baculoviruses was constructed in order to study the influence of downstream ns2a sequences on the processing of the dengue virus ns1 glycoprotein in insect cells. ns1 alone was expressed at a high level in its native dimeric form and processed efficiently through the spodoptera frugiperda (sf) cell secretory pathway. recombinant ns1 was found associated with the plasma membrane and was also secreted into the extracellular medium. although both intra- and extracellular ns1 ...19959049346
effect of methyltransferase inhibitors on the regulation of baculovirus protein synthesis.in the presence of the methyltransferase inhibitor 3-deazaadenosine (3da-ado) the production of infectious autographa californica nuclear polyhedrosis virus (acmnpv) in tissue culture was only slightly affected, while the synthesis of very late proteins (polyhedrin and p10) was abolished. the synthesis of the influenza virus proteins ns1 and ha, expressed under the polyhedrin promoter, was also abolished by 3da-ado. furthermore, 3da-ado interfered with the shut-off of early and late acmnpv prote ...19959049353
a hybrid baculovirus-bacteriophage t7 transient expression system.a hybrid recombinant baculovirus-bacteriophage t7 expression system was developed for transient expression in insect cells of plasmids with foreign genes provided with a t7 promoter. the coding sequence for t7 rna polymerase, with or without a nuclear localization signal, was inserted into the genome of autographa californica nuclear polyhedrosis virus. recombinant viruses stably expressed t7 rna polymerase in insect cells. upon transfection of infected insect cells with plasmids containing the ...19959634768
transovarian transmission of a foreign gene in the silkworm, bombyx mori, by autographa californica nuclear polyhedrosis virus.we introduced a firefly luciferase gene, expressed under control of drosophila heat shock protein gene promoter, into autographa californica nuclear polyhedrosis virus (acnpv). when the 5th instar larvae of the silkworm, bombyx mori, were inoculated with the recombinant virus, luciferase activities were detected in the virus-infected larvae and pupae, and in the newly hatched larvae of the next generation. pcr amplification and southern blot hybridization analysis demonstrated that the luciferas ...19959636278
eukaryotic virus display: engineering the major surface glycoprotein of the autographa californica nuclear polyhedrosis virus (acnpv) for the presentation of foreign proteins on the virus surface.we describe the development of the baculovirus autographa californica nuclear polyhedrosis virus (acnpv) as a vector for the display of distinct proteins on the viral surface in a manner that is analogous to the established bacterial "phage display" systems. as a model system, the marker gene encoding the 26kda protein glutathione-s-transferase (gst) was used to construct several fusions with the major baculovirus glycoprotein gp64 gene. following expression in spodoptera frugiperda (sf9) cells, ...19959636281
identification of a membrane fusion domain and an oligomerization domain in the baculovirus gp64 envelope fusion protein.the baculovirus gp64 envelope fusion protein (gp64 efp) is the major envelope glycoprotein of the budded virion and has been shown to mediate acid-triggered membrane fusion both in virions and when expressed alone in transfected cells. using site-directed mutagenesis and functional assays for oligomerization, transport, and membrane fusion, we localized two functional domains of gp64 efp. to identify a fusion domain in the gp64 efp of the orgyia pseudotsugata multiple nuclear polyhedrosis virus ...19957533858
influence of codon usage and translational initiation codon context in the acnpv-based expression system: computer analysis using homologous and heterologous genes.codon usage by all the known gene sequences from autographa californica nuclear polyhedrosis virus (acnpv) was compared with that of firefly luciferase (luc) and the beta subunit of human chorionic gonadotropin (beta hcg) expressed to contrasting levels in the baculovirus system. the highly expressed luc gene showed a codon usage similar to acnpv genes, as reflected by a very low d-squared statistic value (0.78) and a similar g/c usage (45%) at wobble positions. however, the underexpressed beta ...19957537424
isolation of p10 gene from bombyx mori nuclear polyhedrosis virus and study of its promoter activity in recombinant baculovirus vector system.a homologue of autographa californica npv (acnpv) p10 gene was identified and cloned from bombyx mori npv (bmnpv). bmnpv p10 gene encodes truncated protein of 70 amino acid residues that lacks carboxyl terminus comparing with the p10 protein encoded by acnpv. the putative tata box sequence and the ataag motif which is the consensus sequence of baculovirus very late promoter were conserved. a transfer vector, pbnt1, which includes the p10 promoter region of bmnpv for foreign gene expression was c ...19957547036
expression of human plasminogen activator inhibitor-2 in the baculovirus expression system.a cdna coding for human plasminogen activator inhibitor-2 (pai-2) was transferred into the genome of autographa californica nuclear polyhedrosis virus adjacent to the polyhedrin promoter. cells infected with recombinant virus synthesized a 43-kda pai-2 protein, the majority of which was secreted. the recombinant pai-2 was identical to native pai-2 purified from human placenta with respect to interaction with polyclonal antibody and inhibition of urokinase-type plasminogen activator.19957549934
production of functional human bone morphogenetic protein-2 using a baculovirus/sf-9 insect cell system.a clone of a human bone morphogenetic protein-2 (hbmp-2) cdna was obtained from a cdna library established from human dental pulp cells. after subcloning hbmp-2 cdna into autographa californica nuclear polyhedrosis virus, the recombinant baculovirus was transfected to sf-9 cells. immuno-reactive recombinant hbmp-2 (rhbmp-2) was detected by a polyclonal antibody against xenopus bmp-2 in the transfected insect cells but not in the culture media. three days after treatment with the lysate of the tr ...19957549938
biochemical analysis of the n-glycosylation pathway in baculovirus-infected lepidopteran insect cells.the baculovirus-insect cell system is used routinely for foreign glycoprotein production, but the precise nature of the n-glycosylation pathway in this system remains unclear. some studies indicate that these cells cannot process n-linked oligosaccharides to complex forms containing outer-chain galactose and sialic acid, while others indicate that they can. in this study, we used the major virion envelope glycoprotein of the baculovirus autographa california multicapsid nuclear polyhedrosis viru ...19957571420
replication of orgyia pseudotsugata baculovirus dna: lef-2 and ie-1 are essential and ie-2, p34, and op-iap are stimulatory genes.a transient dna replication assay was used to identify genes located within m.u. 90.5-7.0 of the orgyia pseudotsugata multinucleocapsid nuclear polyhedrosis virus (opmnpv) genome that influenced replication of a reporter plasmid containing an opmnpv origin of replication, when cotransfected into uninfected lymantria dispar cells. the viral transactivator ie-1 and a 2.4-kb subclone were found to be essential for replication. the 2.4-kb region was sequenced and open reading frames were identified. ...19957571435
identification and preliminary characterization of a chitinase gene in the autographa californica nuclear polyhedrosis virus genome.a functional chitinase gene (chia) has been identified in the genome of the autographa californica nuclear polyhedrosis virus (acmnpv). it is expressed in the late phase of virus replication in insect cells. high levels of both endo- and exochitinase activity were detected by 12 hr p.i. and remained stable throughout infection. an acmnpv chia protein-specific antibody was prepared using recombinant material prepared in bacteria. this was used to demonstrate that a product of approximately 58 kda ...19957571437
characterization, sequencing and phylogeny of the ecdysteroid udp-glucosyltransferase gene from two distinct nuclear polyhedrosis viruses isolated from choristoneura fumiferana.the ecdysteroid udp-glucosyltransferase (egt) gene isolated from a plaque-purified isolate of choristoneura fumiferana multinucleocapsid nuclear polyhedrosis virus (cfmnpv) was compared to its homologue from a defective mnpv virus (cfdef) present in wild type virus populations infecting the eastern spruce budworm, c. fumiferana. the egt genes were located in the same relative position within the virus genomes and their genomic location and arrangement were similar to that found in autographa cal ...19957595348
molecular structure and function of cd4 on murine egg plasma membrane.in the present study, the expression of the cd4 molecule on murine egg plasma membrane was confirmed by the indirect immunofluorescence (iif) method. the full-length cd4 cdna from murine eggs was synthesised by the reverse transcriptase-polymerase chain reaction (rt-pcr) method and its authenticity verified by southern blot hybridisation using an end-labelled internal oligonucleotide. the results of dna sequencing showed that the nucleotide sequence of the cdna of cd4 from murine egg mrna was id ...19957613876
identification, sequence, and transcriptional analysis of lef-3, a gene essential for orgyia pseudotsugata baculovirus dna replication.using a transient complementation assay for dna replication of an origin-containing reporter plasmid, a 2.4-kb region (m.u. 48.3-50.1) of the orgyia pseudotsugata multinucleocapsid nuclear polyhedrosis virus (opmnpv) genome was identified that contains a gene essential for opmnpv dna replication. this region was sequenced and open reading frames were identified. replication assays using subclones from this region identified a gene called late expression factor 3 (lef-3), as the essential replica ...19957618274
secretion of active human lecithin-cholesterol acyltransferase by insect cells infected with a recombinant baculovirus.difficulties in purifying the plasma enzyme lecithin-cholesterol acyltransferase (lcat) have hampered detailed studies of its (patho)physiological role in lipoprotein metabolism and of structure-function relationships. potentially, baculovirus-driven expression systems offer a powerful means to produce significant amounts of lcat. accordingly, full-length lcat cdna was cloned into pvl 1392, a high-level expression derivative of autographa californica nuclear polyhedrosis virus (acnpv), and the r ...19957619064
heterologous expression and enzymatic properties of a selenium-independent glutathione peroxidase from the parasitic nematode brugia pahangi.a full-length cdna from the parasitic nematode brugia pahangi encoding a secreted homolog of glutathione peroxidase in which the codon for the active site selenocysteine is substituted naturally by a cysteine codon has been expressed in spodoptera frugiperda (insect) cells via autographa californica nuclear polyhedrosis virus (baculovirus). the recombinant protein was glycosylated and secreted from the cells in tetrameric form. the purified protein showed glutathione peroxidase activity with a r ...19957629152
evaluation of protein-a linked monoclonal antibody latex agglutination test for diagnosis of nuclear polyhedrosis virus (bmnpv) of silkworm bombyx mori l.the symptomology of bmnpv infection in bombyx mori l depends on the stages of infection. discernible symptoms develop at later stages of infection, which leads to improper diagnosis and poor crop yield with sericulturists. in the present study development of direct and protein-a linked monoclonal antibody latex (palmal) agglutination test for the detection of bmnpv infection in silkworm is described. latex beads were precoated with protein-a and then sensitised with monoclonal antibody ma-231 (1 ...19957629276
construction and adhesive properties of a soluble madcam-1-fc chimera expressed in a baculovirus system: phylogenetic conservation of receptor-ligand interaction.madcam-1 is a high endothelial venule adhesion molecule composed of immunoglobulin and mucin-like domains which binds the leucocyte integrin lpam-1 (alpha 4 beta 7), and is largely responsible for the selective homing of lymphocytes to mucosal tissues. a novel soluble form of mouse madcam-1 which is normally membrane bound has been produced by joining the extracellular region of the receptor to the fc domain of human igg1. the madcam-1-fc cdna was inserted into the genome of autographa californi ...19957631157
baculovirus-driven expression and purification of glycine receptor alpha 1 homo-oligomers.the glycine receptor is a ligand-gated anion channel protein of postsynaptic membranes. we expressed a homo-oligomeric receptor composed of human alpha 1 subunits in spodoptera frugiperda cells by infection with a recombinant autographa californica nuclear polyhedrosis virus. a substantial fraction of the recombinant receptor was incorporated as a functional channel protein into the cell's plasma membrane at expression levels 4- to 30-fold higher than in other eukaryotic heterologous expression ...19957635206
expression of pe38 and ie2, viral members of the c3hc4 finger family, during baculovirus infection: pe38 and ie2 localize to distinct nuclear regions.the pe38 gene of autographa californica nuclear polyhedrosis virus represents one of the major early transcripts after viral infection. the function of the pe38 protein, which contains a c3hc4 zinc finger motif, is still not understood. we have raised polyclonal antiserum against the pe38 protein, pe38, produced in bacteria to investigate pe38 expression in the course of infection. a approximately 38-kda polypeptide is first detectable at 2 h postinfection and decreases rapidly after 24 h. durin ...19957636971
transcriptional enhancer activity of hr5 requires dual-palindrome half sites that mediate binding of a dimeric form of the baculovirus transregulator ie1.the hr5 enhancer element stimulates early viral transcription and may function as an origin of dna replication for autographa californica nuclear polyhedrosis virus (acmnpv). the smallest functional unit of hr5 is a 28-bp repeat consisting of an imperfect palindrome (28-mer). to identify essential sequences and examine the molecular basis of hr5 activity, the effects of site-directed mutations on transcriptional enhancement by the 28-mer and binding of the acmnpv transregulator ie1 were investig ...19957636981
baculovirus-mediated production of the human growth hormone in larvae of the silkworm, bombyx mori.to express the cdna encoding human growth hormone (hgh) in larvae of bombyx mori, b. mori nuclear polyhedrosis virus (bmnpv) was employed as an expression vector. for the construction of the recombinant virus, the hgh cdna was inserted into the downstream of the strong polyhedrin promoter to achieve a high level expression. immunoblot analysis revealed that the virus-mediated hgh was synthesized in the larvae and secreted into the hemolymph. the yield of the recombinant hgh synthesized in the la ...19957646491
differential requirements for baculovirus late expression factor genes in two cell lines.a plasmid library of 18 late expression factor (lef) genes (lef library) from the baculovirus autographa californica nuclear polyhedrosis virus (acmnpv) supports transient expression from a late viral promoter in the sf-21 cell line, derived from spodoptera frugiperda. we found, however, that this lef library was unable to support expression from the same promoter in the tn-368 cell line, derived from trichoplusia ni, which is also permissive for acmnpv replication. to identify the additional fa ...19957666527
baculovirus mediated high-level expression of luciferase in silkworm cells and larvae.bombyx mori nuclear polyhedrosis virus (bmnpv)-based baculovirus expression system exploits silkworm larvae as an economical alternative to large-scale cell cultures for production of biomolecules. to generate recombinant bmnpv at high frequencies, we have achieved high efficiency transfection of b. mori cells, bmn, through lipofection. optimal conditions for lipofection were standardized by quantification of the transient expression level of firefly luciferase (luc) reporter gene under control ...19957669303
efficient gene transfer into human hepatocytes by baculovirus vectors.viral vectors are the most efficient tools for gene delivery, and the search for tissue-specific infecting viruses is important for the development of in vivo gene therapy strategies. the baculovirus autographa californica nuclear polyhedrosis virus is widely used as a vector for expression of foreign genes in insect cells, and its host specificity is supposed to be restricted to arthropods. here we demonstrate that recombinant a. californica nuclear polyhedrosis virus is efficiently taken up by ...19957479733
beet western yellows luteovirus capsid proteins produced by recombinant baculoviruses assemble into virion-like particles in cells and larvae of bombyx mori.we subcloned various constructs of the beet western yellows luteovirus (bwyv) 3' proximal genes into bombyx mori nuclear polyhedrosis virus (bmnpv) transfer vectors and constructed recombinant bmnpvs. the recombinant bmnpvs were used to infect bmn cells and b. mori larvae. protein expression was analyzed by sds-page and immunoblot analysis, and the bwyv-encoded capsid protein (cp), p19, readthrough protein (rt), and a modified capsid-readthrough protein (cp-rt*) were detected in bmn cells infect ...19957483264
functional expression of an alpha anti-insect scorpion neurotoxin in insect cells and lepidopterous larvae.the leiurus quinquestriatus hebraeus alpha anti-insect toxin (lqh alpha it) cdna was engineered into the autographa californica nuclear polyhedrosis virus (acnpv) genome. insect cells infected with the recombinant virus secreted a functional lqh alpha it polypeptide. spodoptera littoralis and heliothis armigera larvae injected with recombinant budded virus, showed typical intoxication symptoms. this recombinant virus showed enhanced insecticidal potency against h. armigera larvae compared with w ...19957498537
transcriptional regulation of cell line-dependent, baculovirus-mediated expression of foreign genes.cell line-dependent expression of foreign genes in the baculovirus system was investigated using a recombinant vac beta hcg-luc virus carrying two reporter genes--beta subunit of human chorionic gonadotropin (beta hcg) and luciferase (luc)--placed under the transcriptional control of the autographa californica nuclear polyhedrosis virus (acnpv) polyhedrin gene promoters. five different lepidopteran cell lines derived from spodoptera frugiperda (sf21 and sf9), bombyx mori (bmn and bm5), and trich ...19957530452
use of juvenile hormone esterase as a novel reporter enzyme in the baculovirus expression system.juvenile hormone esterase (jhe) has a number of characteristics favorable for use as a reporter enzyme. it is extremely stable under a variety of adverse conditions including in organic solvent. jhe is easily detected by a rapid and sensitive colorimetric assay, and detection is facilitated by export of the enzyme from the cell. its use is illustrated in the baculovirus expression system by promoter studies and evaluation of culture conditions necessary for optimal production of recombinant prot ...19957730431
characterization of v-cath, a cathepsin l-like proteinase expressed by the baculovirus autographa californica multiple nuclear polyhedrosis virus.autographa californica multiple nuclear polyhedrosis virus (acmnpv) contains a 966 bp orf that encodes a papain type cysteine proteinase with cathepsin l-like characteristics. using western blot analysis of infected cell extracts we showed that v-cath proteinase has 35.5 kda and 32 kda precursor forms which are processed to a 27.5 kda mature form in a manner characteristic of papain and cathepsin l. v-cath proteinase activity was greatest under acidic conditions (ph 5.0) and was reduced in the p ...19957730794
overexpression of bombyx mori prothoracicotropic hormone using baculovirus vectors.recombinant baculoviruses were constructed that express the cdna encoding the prothoracicotropic hormone (ptth) of bombyx mori. this hormone stimulates the production of ecdysteroids by the insect's prothoracic glands. two groups of viruses were constructed, expressing either the entire cdna encoding prepro-ptth, or a synthetic chimeric gene encoding a signal peptide fused to the mature ptth subunit. in both cases, the genes were expressed in wild-type autographa californica nuclear polyhedrosis ...19957742834
the cagt motif functions as an initiator element during early transcription of the baculovirus transregulator ie-1.the highly conserved tetranucleotide cagt is located at the rna start site of the transregulator gene ie-1 of autographa californica nuclear polyhedrosis virus (acmnpv). the presence of this motif within numerous baculovirus early promoters and its similarity to transcriptional initiators suggested a fundamental role in viral transcription regulation. to determine the function of the cagt motif, site-specific mutations were introduced within the ie-1 promoter fused to a reporter gene within acmn ...19957745705
the lef-3 gene of autographa californica nuclear polyhedrosis virus encodes a single-stranded dna-binding protein.the autographa californica nuclear polyhedrosis virus (acnpv) replicates in the nuclei of infected cells and encodes several proteins required for viral dna replication. as a first step in the functional characterization of viral replication proteins, we purified a single-stranded dna-binding protein (ssb) from acnpv-infected insect cells. nuclear extracts were chromatographed on single-stranded dna agarose columns. an abundant protein with an apparent molecular weight of 43,000 was eluted from ...19957745748
the autographa californica nuclear polyhedrosis virus homologous region 1a: identical sequences are essential for dna replication activity and transcriptional enhancer function.hr1a consists of two 30-bp imperfect palindrome sequences separated by 58 bp and each palindrome contains a naturally occurring ecori site at its core. plasmid subclones of the hr1a-containing acmnpv hindiii-n fragment were examined for their ability to replicate in virus-infected (spodoptera frugiperda) sf9 cells, and to stimulate transcription when linked in cis with a 39k gene promoter-beta-glucuronidase fusion and cotransfected into cells along with a plasmid (ple-1) containing the gene enco ...19957747446
a temperature-sensitive ie1 protein of autographa californica nuclear polyhedrosis virus has altered transactivation and dna binding activities.the temperature-sensitive mutant tsb821 of autographa californica nuclear polyhedrosis virus has two nucleotide transitions in the transregulatory ie1 gene (ribiero et al. (1994) j. virol. 68, 1075-1084). these mutations result in two conservative amino acid substitutions in the encoded protein. to determine whether the mutations affect the ability of ie1 to interact with viral enhancer elements, electrophoretic mobility shift assays were performed using extracts prepared from infected cells and ...19957747488
expression of the ie1 transactivator of autographa californica nuclear polyhedrosis virus during viral infection.the immediate-early ie1 protein of autographa californica nuclear polyhedrosis virus (acmnpv) is an important regulator of viral gene transcription. to provide a tool for further analysis of the expression and function of ie1, a polyclonal antiserum was raised against ie1 expressed in bacteria. immunoblot analysis of infected cell lysates was used to monitor the accumulation of ie1 throughout the viral life cycle. when extracts were prepared in the presence of phosphatase inhibitors, only one pr ...19957747489
synthesis and processing of the rubella virus p110 polyprotein precursor in baculovirus-infected spodoptera frugiperda cells.in order to study the processing of rubella virus (rv) structural proteins (capsid protein, of 33 kda; e2 of 42-47 kda; and e1 of 58 kda) in spodoptera frugiperda (fall armyworm) cells, a 24s cdna encoding the polyprotein precursor, p110, was inserted under the transcriptional regulation of the polyhedrin gene promoter of the autographa californica nuclear polyhedrosis virus (acnpv) and expressed during viral infection. by immunoblot analysis using antibodies directed against whole rv and the in ...19957754676
high level synthesis and secretion of human urokinase using a late gene promoter of the autographa californica nuclear polyhedrosis virus.a cdna encoding human urokinase was inserted into the autographa californica nuclear polyhedrosis virus (acnpv) genome at the polyhedrin gene locus under control of a duplicated copy of the late, basic protein gene promoter. the insect-derived urokinase was produced predominantly in the form of single-chain, pro-urokinase, with a molecular mass of 50 kda, and demonstrated fibrinolytic activity. synthesis and secretion of urokinase was first detected at 6 hours post-infection and continued steadi ...19957766008
deletion of the baculovirus ecdysteroid udp-glucosyltransferase gene induces early degeneration of malpighian tubules in infected insects.deletion of the ecdysteroid udp-glucosyltransferase gene (egt) from the autographa californica nuclear polyhedrosis virus (acnpv) genome increases the speed of killing of this virus (d. r. o'reilly and l. k. miller, bio/technology 9:1086-1089, 1991). second-instar spodoptera exigua larvae are killed more rapidly by the egt deletion mutant of acnpv than by wild-type acnpv. unlike wild-type acnpv-infected larvae, larvae infected with an egt deletion mutant molt and resume feeding as mock-infected ...19957769717
properties of a baculovirus mutant defective in the protein phosphatase gene.autographa california nuclear polyhedrosis virus (acmnpv) contains a gene, ptp, encoding a protein tyrosine/serine phosphatase, bv-ptp. to investigate the biological function of ptp in the baculoviral replication cycle, we constructed a recombinant baculovirus, vptpdel, in which the catalytically active site of bv-ptp was deleted. although the vptpdel mutant was viable in cell culture, it was partially defective in occluded virus production in sf-21 but not tn-368 cell lines. sf-21 cells infecte ...19957769718
the baculovirus transactivator ie1 binds to viral enhancer elements in the absence of insect cell factors.the transregulatory ie1 protein of autographa californica nuclear polyhedrosis virus binds to the viral enhancer element hr5. to test whether ie1 binds independently of host cell factors, ie1 was translated in rabbit reticulocyte extracts and tested for dna binding activity by an electrophoretic mobility shift assay. complexes with the hr5 probe were detected with translation reaction mixtures primed with ie1 rna but not with control translation reaction mixtures. however, the dna-protein comple ...19957769721
n-glycosylated polypeptides synthesized in the early phase of acnpv infected insect cells.infected cell specific polypeptides (icsp) in autographa californica nuclear polyhedrosis virus (acnpv) infected spodoptera frugiperda cell line, sf9, were radiolabelled with [14c] mannose and the labelled proteins were monitored at various time intervals. a major glycoprotein of molecular size of 64 kda appears as early as 3h post infection (p.i.) which peaks at 12h p.i., but in presence of tunicamycin this protein is not labelled. two other glycoproteins of molecular size 67 kda and 63 kda app ...19957773189
developmental resistance in fourth instar trichoplusia ni orally inoculated with autographa californica m nuclear polyhedrosis virus.larvae of lepidopteran insects commonly become increasingly resistant to baculovirus infections as they age. the mechanism responsible for this development resistance is not known, but the phenomenon does not occur if the viral inoculum is administered intrahemocoelically instead of orally, which is the natural route of infection. this observation indicates that the factors mediating developmental resistance are operative during infection of the primary target tissue, the larval midgut, and not ...19957778273
identification and analysis of a putative origin of dna replication in the choristoneura fumiferana multinucleocapsid nuclear polyhedrosis virus genome.a recombinant plasmid containing the choristoneura fumiferana multinucleocapsid nuclear polyhedrosis virus (cfmnpv) hindiii r fragment (m.u. 2.2-3.9) was shown to undergo cfmnpv infection-dependent dna replication in cf-124t cells. replication of this dna sequence was detectable by 24 hr p.i. and did not appear to have resulted as a consequence of recombination with the virus genome. replication was inhibited by mimosine, an inhibitor of eukaryotic dna replication. these data suggest that hindii ...19957778276
the wild-type autographa californica nuclear polyhedrosis virus induces apoptosis of spodoptera littoralis cells.spodoptera littoralis cells infected with the autographa californica multiple nuclear polyhedrosis virus (acmnpv) yielded significantly lower budded virus titers than spodoptera frugiperda-infected cells and produced very low levels of polyhedrin. relative to acmnpv-infected s. frugiperda sf9 cells viral dna replication was severely reduced in spodoptera littoralis sl2 cells. microscopic examination of sl2-infected cells revealed progressive cell blebbing starting at 6-8 hr postinfection and cul ...19957778284
identification, localization, transcription, and sequence analysis of the choristoneura fumiferana nuclear polyhedrosis virus dna polymerase gene.the location of the choristoneura fumiferana baculovirus dna polymerase gene was determined by hybridization analysis using a probe prepared from the previously identified polymerase gene from the autographa californica multiple nuclear polyhedrosis virus. dna sequence analysis revealed that the choristoneura fumiferana baculovirus dna polymerase gene consists of 2970 base pairs encoding 990 amino acids (114.2 kda). transcriptional analysis demonstrated that overlapping transcripts of 3.2 and 4. ...19957778286
comparative pathogenesis of autographa californica m nuclear polyhedrosis virus in larvae of trichoplusia ni and heliothis virescens.we compared early viral pathogenesis and dose-mortality relationships for larvae of two highly susceptible hosts, trichoplusia ni and heliothis virescens, using a construct of acmnpv containing the lacz reporter gene. larvae were inoculated either as newly molted fourth instars (4(0)) or 15 hr after the molt (4(15)). in 4(0)-inoculated larvae, first lacz expression was detected in the midgut epithelium of t. ni at 4 hr postinoculation (hpi) compared to 18 hpi in h. virescens, and systemic infect ...19957778288
the baculovirus gp64 envelope fusion protein: synthesis, oligomerization, and processing.the baculovirus gp64 envelope fusion protein (gp64 efp) is a class i integral membrane protein that enters the secretory pathway and is oligomerized and extensively processed during transport to the plasma membrane. the kinetics of gp64 efp biosynthesis, oligomerization, and processing in orgyia pseudotsugata multicapsid nuclear polyhedrosis virus (opmnpv)-infected lymantria dispar cells were examined by pulse label, pulse-chase, and immunoprecipitation experiments. relative rates of gp64 efp sy ...19957778291
nucleotide sequence and transcriptional analysis of the gp41 gene of spodoptera frugiperda nuclear polyhedrosis virus.the spodoptera frugiperda multiple nucleocapsid nuclear polyhedrosis virus (sfmnpv) gp41 structural protein gene was located in the 1.9 kbp ecori-s fragment and sequenced. an open reading frame (orf) of 999 nucleotides was detected that encoded a protein of 332 amino acids. the gp41 gene transcript was detected after 12 h post-infection (p.i.) and remained detectable at 48 h p.i. two major mrnas, about 1.6 and 2.8 kb in length, were determined by northern blot analysis. primer extension analysis ...19957782772
mutations in the autographa californica multinucleocapsid nuclear polyhedrosis virus 25 kda protein gene result in reduced virion occlusion, altered intranuclear envelopment and enhanced virus production.serial passage of nuclear polyhedrosis viruses (npvs) through cultured cell lines results in the appearance of mutants with a complex phenotype referred to as the 'few polyhedra' (fp) phenotype. the altered plaque morphology and reduced occlusion production associated with the fp phenotype have been observed in autographa californica multinucleocapsid nuclear polyhedrosis virus (acmnpv) bearing mutations in the gene encoding the 25 kda protein (25k gene). in this study, we sequenced the 25k gene ...19957782773
p10 genes of bombyx mori nuclear polyhedrosis virus and autographa californica multiple nuclear polyhedrosis virus.ecor i-p fragment has been cloned from autographa californica multiple nuclear polyhedrosis virus (acmnpv) genomic dna and used as a probe. 0.5-kb and 1.1-kb fragments including p10 gene from bombyx mori nuclear polyhedrosis virus (bmnpv) have been hybridized. the p10 orf was located in the ecor i-r fragment. initiation codon atg of p10 from bmnpv has been mutated by pcr, and the atg region became a bgl ii site. a novel transfer vector pbmacpv-1 has been constructed using both the p10 5'-flankin ...19957695816
eighteen baculovirus genes, including lef-11, p35, 39k, and p47, support late gene expression.we report the identification of four additional genes of the autographa californica nuclear polyhedrosis virus involved in expression from a late baculovirus promoter in transient expression assays. three of these genes, p35, 39k, and p47, have been previously described. the role of the p35 gene product in late gene expression may be related to its ability to block apoptosis, since two other baculovirus genes also known to block apoptosis, cp-iap and op-iap, were able to functionally replace p35 ...19957815564
the roles of eighteen baculovirus late expression factor genes in transcription and dna replication.a set of 18 plasmid subclones of the autographa californica nuclear polyhedrosis virus genome supports expression from a late viral promoter in transient expression assays (j. w. todd, a. l. passarelli, and l. k. miller, j. virol. 69:968-974, 1995). using this set of plasmids, we have assigned a role for each of the 18 genes required for optimal late gene expression with respect to its involvement at the levels of transcription, translation, and/or dna replication. rnase protection analyses demo ...19957815565
expression and functional analysis of a baculovirus gene encoding a truncated protein kinase homolog.autographa californica nuclear polyhedrosis virus (acmnpv) potentially encodes a 215-amino acid polypeptide containing 6 out of 11 motifs conserved among eukaryotic protein kinases (morris et al., virology 200, 360-369, 1994). we examined the expression of this gene, named pk2, at the transcriptional and translational levels and the possible role of the gene during baculovirus replication in cell culture and insect larvae. northern (rna) blot analysis revealed that pk2 was transcribed primarily ...19957831787
nucleotide sequence and transcriptional analysis of the dna polymerase gene of bombyx mori nuclear polyhedrosis virus.a gene encoding a putative dna polymerase (pol) of bombyx mori nuclear polyhedrosis virus (bmsnpv) was cloned and sequenced. the gene included an open reading frame (orf) encoding a polypeptide of 988 amino acids with a predicted molecular mass of 114.65 kda. the deduced amino acid sequence of the bmsnpv pol orf showed an overall identity of 96 and 45% to those of the autographa californica npv (acmnpv) pol orf and the lymantria dispar npv pol orf, respectively, and contained sequences conserved ...19957831799
ubiquitin is attached to membranes of baculovirus particles by a novel type of phospholipid anchor.purified budded virions of autographa californica nuclear polyhedrosis virus (acnpv) contain abundant amounts of free ubiquitin, which has an altered electrophoretic mobility on sds gels as compared with standard ubiquitin. phase extraction of virion proteins with triton x-114 indicated that the modified form of ubiquitin behaved as an integral membrane protein. the membrane-bound form of ubiquitin was labeled with both phosphate and palmitate, and its electrophoretic mobility was altered by tre ...19957834750
sequence analysis and transcriptional mapping of the orf-2 gene of autographa californica nuclear polyhedrosis virus.sequencing of the dnapol promoter region of autographa californica nuclear polyhedrosis virus (acnpv) revealed an overlapping open reading frame (orf) in an antisense orientation, referred to as orf-2. analysis of the orf-2 deduced amino-acid sequence revealed two short regions of homology with a similar orf from lymantria dispar nuclear polyhedrosis virus (ldnpv). two 3' processing signals of this gene, expressed late during infection, were shown to be located on the orf-2 stop codon and 162 nu ...19957835700
proteolysis of p6.9 induced by cytochalasin d in autographa californica m nuclear polyhedrosis virus-infected cells.cytochalasin d, a fungus-derived compound that interferes with actin polymerization, inhibits autographa californica m nuclear polyhedrosis virus production in infected spodoptera frugiperda (iplb-sf-21) cells. cytochalasin d appears to inhibit nucleocapsid morphogenesis by interfering with nucleoprotein packaging. we were interested in determining, therefore, whether the drug affected the synthesis or processing of p6.9, the major core protein involved in nucleoprotein packaging. we found that ...19957871717
identification of essential trans-acting regions required for dna replication of the orgyia pseudotsugata multinucleocapsid nuclear polyhedrosis virus: lef-1 is an essential replication gene.a transient replication assay for the identification of baculovirus genes that are essential for replication of an origin-containing reporter plasmid was established for the orgyia pseudotsugata multinucleocapsid nuclear polyhedrosis virus (opmnpv). using a replication origin located on the opmnpv hindiii-n fragment, we identified a subset of cosmids and plasmids from an opmnpv cosmid library that was able to supply all the essential trans-acting factors and support replication of the origin-con ...19957886946
identification and characterization of a second putative origin of dna replication in a baculovirus of orgyia pseudotsugata.a 7.5-kb region (96.8-2.5 m.u.), called op5, of the orgyia pseudotsugata multinucleocapsid nuclear polyhedrosis virus (opmnpv) genome that contains an origin of dna replication was characterized. this region replicates several times more efficiently and is unrelated to the previously identified putative origin of replication located on the viral hindiii-n fragment. in contrast to hindiii-n, the origin on op5 contains repeated sequences with limited sequence identity to the homologous regions fro ...19957886962
expression of the human interleukin-2 receptor gamma chain in insect cells using a baculovirus expression vector.the gene encoding the gamma-chain of the human interleukin-2 receptor was expressed in lepidopteran insect cells using the baculovirus expression vector system. the corresponding gene was inserted under the polyhedrin promoter of the autographa californica nuclear polyhedrosis virus and expressed in the spodoptera frugiperda insect cell line sf9 during viral infection. the recombinant receptor protein was identified by immunoblotting in cell lysates, prepared from insect cells infected with the ...19957899821
baculovirus expression of human basic fibroblast growth factor from a synthetic gene: role of the kozak consensus and comparison with bacterial expression.synthetic genes encoding the 146 and 155 amino acid forms of human basic fibroblast growth factor (bfgf) were constructed with codon usage biased towards the polyhedrin-encoding gene of autographa californica nuclear polyhedrosis virus (acnpv). expression of both bfgf genes in spodoptera frugiperda (sf-21) suspension cell culture using a recombinant baculovirus yielded approximately 2.5 mg of mitogenically fully active protein per 10(9) cells following heparin-affinity chromatography. to improve ...19957999789
functional dissection of the ie2 gene product of the baculovirus autographa californica nuclear polyhedrosis virus.the psti-n genomic fragment of autographa californica nuclear polyhedrosis virus (acnpv) encodes an immediate-early protein, ie2, that functions as a promiscuous transactivator of other early viral promoters and heterologous promoters both in vivo and in vitro. ie2 contains several sequence motifs that are common to transcriptional activators. we have employed site-directed mutagenesis coupled with transient-expression assays to identify the amino acid sequences of ie2 that are essential for tra ...19948009830
autographa californica nuclear polyhedrosis virus, pdv, and ecv viral envelopes and nucleocapsids: structural proteins, antigens, lipid and fatty acid profiles.autographa californica nuclear polyhedrosis virus (acmnpv) infection results in the production of two types of infectious, enveloped viruses. as both of these viral forms play significantly different roles in the virus life cycle, the different functional characteristics in the roles of the virus may be explained, in part, by the protein and lipid composition and source of the viral envelopes. both viruses utilize different maturation and envelopment strategies: extracellular virus (ecv) obtains ...19948009843
reiterated dna fragments in defective genomes of autographa californica nuclear polyhedrosis virus are competent for acmnpv-dependent dna replication.we previously reported on the generation of approximately 50-kb size defective genomes (dgs) which appeared to retain less than 2.2% of the standard autographa californica nuclear polyhedrosis virus (acmnpv) dna between 85.0 and 87.2 mu while the rest of the virus dna had been largely deleted (lee and krell, j. virol., 66:4339-4347, 1992). to investigate these presumably repeated sequences further, we cloned and analyzed the most abundant hypermolar 1.80-kb xhol dna fragment as well as a minor b ...19948009853
engineering the assembly pathway of the baculovirus-insect cell expression system.the synthesis of complex biological structures such as antibodies using recombinant dna technology is a major biotechnological advance. active murine antibody (igg) oligomers, composed of two heavy (h) and two light (l) polypeptide chains, have been expressed and secreted by the baculovirus-insect cell expression system. unfortunately, expression of the functional antibodies is accompanied by the formation of abnormal protein complexes and aggregates in which the polypeptide chains are bound tog ...19948010671
expression of aleutian mink disease parvovirus capsid proteins in a baculovirus expression system for potential diagnostic use.a 2.3-kb cdna clone encoding aleutian mink disease parvovirus (adv) structural proteins vp1 and vp2 was inserted into the polyhedron gene of autographa californica nuclear polyhedrosis virus (acnpv) and expressed by the recombinant virus, acadv-1, in spodoptera frugiperda-9 cells. sodium dodecyl sulfate-polyacrylamide gel electrophoresis and western immunoblot analysis (wia) indicated that synthesis of both vp1 and vp2 was being directed by acadv-1. fluorescence microscopic examination of acadv- ...19948011776
superior expression of juvenile hormone esterase and beta-galactosidase from the basic protein promoter of autographa californica nuclear polyhedrosis virus compared to the p10 protein and polyhedrin promoters.the expression characteristics of the p10, polyhedrin and basic protein promoters of autographa californica nuclear polyhedrosis virus were compared using two reporter enzymes, juvenile hormone esterase (jhe) and beta-galactosidase. in these systems, jhe is exported from the cell and beta-galactosidase is localized to the cytosol. expression of jhe from the basic, p10 and polyhedrin promoters was first detected in the medium at 13, 19 and 27 h post-infection respectively. the basic protein promo ...19948021586
identification of the gp67 gene of a baculovirus pathogenic to the spruce budworm, choristoneura fumiferana multinucleocapsid nuclear polyhedrosis virus.the baculovirus gp67 gene encodes a ph-dependent membrane fusion protein and has been identified in both autographa californica multinucleocapsid nuclear polyhedrosis virus (acmnpv) and orygia pseudotsugata mnpv (opmnpv). we have identified a homologous gene in the spruce budworm virus, choristoneura fumiferana mnpv (cfmnpv). the cfmnpv gp67 gene is 79% identical to acmnpv gp67 at the level of nucleotide sequence and 82% identical at the level of predicted amino acid sequence. as with opmnpv and ...19948021612
genome organization of the dna-binding protein gene region of cryptophlebia leucotreta granulosis virus is closely related to that of nuclear polyhedrosis viruses.the nucleotide sequence and genomic organization of a conserved genome region within the ecori g fragment of cryptophlebia leucotreta granulosis virus (clgv) is presented. five open reading frames (orfs) were identified which were homologous to those of autographa californica nuclear polyhedrosis virus (acmnpv), located upstream of the helicase gene (p143) at 63.6 to 65.6 map units. the orfs of clgv and acmnpv share nucleotide sequence homologies of about 47 to 53% and are very similarly arrange ...19948021613
the complete dna sequence of autographa californica nuclear polyhedrosis virus.the complete nucleotide sequence of the genome of clone 6 of the baculovirus autographa californica nuclear polyhedrosis virus (acnpv) has been determined. the molecule comprises 133,894 base pairs and has an overall a + t content of 59%. our analysis suggests that the virus encodes some 154 methionine-initiated, and potentially expressed, open reading frames (orfs) of 150 nucleotides or greater. these orfs are distributed evenly throughout the virus genome on either strand. the orfs are arrange ...19948030224
the autographa californica nuclear polyhedrosis virus ie2 gene encodes a transcriptional regulator.the baculovirus autographa californica nuclear polyhedrosis virus (acnpv) expresses several immediate early genes in infected cells. one of these, ie2 (formerly called ien), was mapped and identified by its ability to augment ie1-mediated transactivation of the delayed early 39k gene. ie2 also stimulated the expression of reporter genes linked to the ie1 promoter. to test whether the increase in expression was due to increased mrna, primer extension analyses were performed. spodoptera frugiperda ...19948030237
Refresh
firstprev
12345678910...
nextlast
Displaying items 401 - 500 of 1347