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a baculovirus dual expression vector derived from the autographa californica nuclear polyhedrosis virus polyhedrin and p10 promoters: co-expression of two influenza virus genes in insect cells.a baculovirus transfer vector, pacuw3, was developed to facilitate the insertion of two influenza virus genes, those encoding the haemagglutinin (ha) and neuraminidase (na) membrane glycoproteins, into the autographa californica nuclear polyhedrosis virus genome in a single cotransfection experiment. the na gene was inserted in place of the polyhedrin coding sequences under the control of the polyhedrin promoter, whereas the ha gene was placed under the control of a copy of the p10 promoter at a ...19911765769
structural comparison of the autographa californica nuclear polyhedrosis virus-induced rna polymerase and the three nuclear rna polymerases from the host, spodoptera frugiperda.a partial subunit structure has been determined for the novel rna polymerase that is induced in fall armyworm (spodoptera frugiperda) cells upon infection with the autographa californica nuclear polyhedrosis virus (acnpv). the putative structure includes nine polypeptides; the complexity of this structure is in accord with the high sedimentation coefficient (15s) estimated for this enzyme. a comparison of the putative structure of the virus-induced polymerase with those of the three host nuclear ...19911767583
expression of the s1 sub-unit of pertussis toxin using a recombinant baculovirus.the full-length genome of the s1 sub-unit containing the native signal sequence was inserted downstream of the polyhedrin promoter of autographa californica nuclear polyhedrosis virus. a large amount of 28 kd protein, which was larger than the authentic s1 sub-unit (26 kd), was detected in insect cells infected with the recombinant virus by coomassie blue staining after resolving by sds-page. although the level of expression of the recombinant s1 sub-unit was high, immunoblotting and n-terminal ...19911778337
the nonstructural protein (nss) encoded by the ambisense s rna segment of tomato spotted wilt virus is associated with fibrous structures in infected plant cells.the open reading frame located in the viral strand of the ambisense s rna of tomato spotted wilt virus (tswv), was cloned into transfer vector pac33dz1 and inserted downstream of the polyhedrin promoter in the autographa californica nuclear polyhedrosis virus genome. recombinant baculoviruses were obtained that showed a high-level expression of a 52.4-kda protein corresponding to the inserted tswv gene. the viral protein thus produced was purified and injected into rabbits to raise antibodies. w ...19911826573
secretion of single-chain urokinase-type plasminogen activator from insect cells.a cdna encoding human urokinase-type plasminogen activator was inserted downstream from the polyhedrin promoter of the baculovirus autographa californica nuclear polyhedrosis virus. a protein of similar mr to urokinase (uk) was synthesized and approx. 90% was secreted from recombinant virus-infected spodoptera frugiperda cells. zymography and western blotting analysis of the insect-derived protein demonstrated that it was comprised solely of the high-mr form of uk. no low-mr uk was detected. ami ...19911834526
high-level expression of the epstein-barr virus alkaline deoxyribonuclease using a recombinant baculovirus: application to the diagnosis of nasopharyngeal carcinoma.the epstein-barr virus (ebv) alkaline deoxyribonuclease (dnase) was inserted into the baculovirus autographa californica nuclear polyhedrosis virus (acmnpv). infection of the insect cell line spodoptera frugiperda (sf9) with the recombinant virus led to the expression of an enzymatically active alkaline dnase. the recombinant ebv alkaline dnase was highly soluble, and the recombinant baculovirus produced approximately 10-20 mg of ebv dnase per 1 x 10(9) cells. the recombinant enzyme activity was ...19911847261
detection and analysis of autographa californica nuclear polyhedrosis virus mutants with defective interfering properties.defective interfering particles (dips) were generated upon continuous production of autographa californica nuclear polyhedrosis virus (acnpv) in bioreactors. this configuration mimicked the serial undiluted passaging of virus, which is known to result in plaque-morphology mutants. restriction enzyme analysis of dip-containing preparations of extracellular virus showed the presence of many dna fragments in less than equimolar amounts. these fragments were colinear on the physical map of acnpv and ...19911853572
a baculovirus homolog of a cu/zn superoxide dismutase gene.a gene with greater than 50% amino acid sequence identity to eukaryotic cu/zn superoxide dismutase genes (sod) was found at 19 map units in the genome of the baculovirus autographa californica nuclear polyhedrosis virus (acmnpv). this gene was transcribed into 1.4- and 1.5-kb rnas late in virus infection. the two late rnas have coterminal 3' ends but initiate from two different start points, both of which map to the central adenine of the sequence motif attaag. the late gene product was found to ...19911871962
dengue-1 virus envelope glycoprotein gene expressed in recombinant baculovirus elicits virus-neutralizing antibody in mice and protects them from virus challenge.in order to test the feasibility of baculovirus (autographa californica nuclear polyhedrosis virus, acnpv) expression vectors for making immunogens against dengue-1 (den-1) virus, a portion of the envelope (e) glycoprotein gene of den-1 virus was cloned and expressed. the recombinant baculovirus contains 107 nucleotides from the 3' terminus of the den-1 matrix (m) gene, which encodes a hydrophobic signal peptide and extends through the first 1, 245 nucleotides of e, terminating 243 nucleotides b ...19911877710
synthesis of the virus-specified tubules of epizootic haemorrhagic disease virus using a baculovirus expression system.the formation of virus-specific tubules is one of the most characteristic features in the orbivirus infection cycle, yet little is known about their role in virus replication. the tubuli are composed of a major nonstructural protein, ns1. we have investigated the expression of the ns1-encoding gene of epizootic haemorrhagic disease virus serotype 2 (alberta-strain) by producing a recombinant autographa californica nuclear polyhedrosis virus (acnpv). prior to cloning in the baculovirus transfer v ...19911891959
novel regulatory properties of the ie1 and ie0 transactivators encoded by the baculovirus autographa californica multicapsid nuclear polyhedrosis virus.the baculovirus autographa californica multicapsid nuclear polyhedrosis virus expresses two immediate-early genes from the hindiii-g region (map units 90.4 to 96.8) of the genome. during the early phase of infection, nonspliced 1.9-kb and spliced 2.1-kb transcripts are expressed which encode the ie1 and ie0 (spliced ie1) gene products, respectively. these two gene products differ only in that ie0 contains an additional 54 amino acids at the amino terminus. rna analysis of these two genes during ...19911895384
expression of human alzheimer amyloid precursor protein in insect cells.the amyloid beta-peptide is a major constituent of amyloid deposited in the brains of patients with alzheimer's disease and is derived from a larger precursor protein/s (app-695, 751, 770). a human cdna encoding full-length app-751 was inserted into the genome of autographa californica nuclear polyhedrosis virus under transcriptional regulation of the viral polyhedrin gene promoter. the recombinant virus was used to infect insect cells, which resulted in the abundant expression of app-751. analy ...19911899577
synthesis of biologically active adenovirus preterminal protein in insect cells using a baculovirus vector.a dna fragment encoding the polyhedrin promoter of autographa californica multiple nuclear polyhedrosis virus (acmnpv strain) was constructed using overlapping oligodeoxyribonucleotides (oligos), which included the 5'-untranslated leader sequence of the polyhedrin-encoding gene. this dna fragment was cloned into an intermediate transfer vector (pkx105) providing a unique bamhi site for the insertion of foreign genes. the escherichia coli lacz gene was first cloned at the bamhi site of pkx105 and ...19911905255
alpha-mannosidase-catalyzed trimming of high-mannose glycans in noninfected and baculovirus-infected spodoptera frugiperda cells (iplb-sf-21ae). a possible contributing regulatory mechanism for assembly of complex-type oligosaccharides in infected cells.incubation of a spodoptera frugiperda (iplb-sf-21ae) cell extract with the oligosaccharide man9glcnac2, the aglucosyl derivative of the glycan that is normally transferred from the dolichol carrier to the relevant asn residue in the nascent protein, results in its trimming to man6glcnac2, an intermediate that is relatively stable to further alpha-d-mannosidase action in these cells. on the other hand, incubation of a similar extract from cells that had been infected for various times with a wild ...19911911772
transcriptional analyses of baculovirus polyhedrin and foreign gene expression relative to baculovirus p10 mrna levels.comparisons have been made between the p10 and polyhedrin mrna levels recovered from spodoptera frugiperda cells infected with autographa californica nuclear polyhedrosis virus (acnpv). in molar terms and from 18 h post-infection (p.i.), the polyhedrin mrna species increased to levels one and a half times to twice as high as the p10 levels. the influence of the polyhedrin leader sequence on the expression of a foreign gene under the control of the polyhedrin promoter was investigated using a ser ...19911919531
baculovirus gp64 gene expression: analysis of sequences modulating early transcription and transactivation by ie1.expression of the baculovirus major envelope glycoprotein gene (gp64) is regulated by transcription from both early and late promoters. to characterize the early promoter and identify sequences involved in the regulation of gp64 early transcription, promoter-reporter gene fusions were generated from the orygia pseudotsugata nuclear polyhedrosis virus gp64 promoter and were analyzed by transient expression in uninfected insect cells. for these analyses, 5' deletion mutations were constructed in t ...19911920618
baculovirus-mediated expression of human apolipoprotein e in manduca sexta larvae generates particles that bind to the low density lipoprotein receptor.human apolipoprotein e (apoe) is a ligand for the low density lipoprotein (ldl) receptor and mediates the catabolism of several classes of lipoprotein particles. binding of apoe to the ldl receptor requires association of apoe with lipid in a vesicle or a lipoprotein particle. because of this requirement, purified apoe or apoe derived directly from bacterial expression systems does not bind to the ldl receptor. to overcome this problem and to facilitate analysis of apoe structure, recombinant ba ...19911924311
nucleotide sequence of the autographa californica nuclear polyhedrosis 9.4 kbp ecori-i and -r (polyhedrin gene) region.the nucleotide sequence of a 9.4-kbp region including the polyhedrin gene of the c6 strain of the autographa californica nuclear polyhedrosis virus (acmnpv) genome was determined. these data provide a complete description of the ecori-i fragment, which is used to produce transfer vectors for inserting foreign genes into the acmnpv. ten potential open reading frames (orfs) were identified in the complete sequence, on either strand of dna. the largest of these was 1629 nucleotides in length and wa ...19911926775
high-level secretion of the extracellular domain of the human growth hormone receptor using a baculovirus system.a chemically synthesized gene (hghr-ed) coding for the extracellular domain (ed) of the human growth hormone (hgh) receptor (hghr) was inserted into the genome of autographa californica nuclear polyhedrosis virus adjacent to the polyhedrin promoter. spodoptera frugiperda cells infected with the recombinant virus secreted a protein with hgh-binding activity into the medium. the secreted 35-kda protein was purified to near homogeneity. the purified protein exhibited a high binding affinity (kd = 0 ...19911937047
location and nucleotide sequence of the gene encoding the viral enhancing factor of the trichoplusia ni granulosis virus.the gene encoding the viral enhancing factor (vef) of trichoplusia ni granulosis virus has been cloned from a lambda gt11 expression library, and the complete nucleotide sequence determined. the vef gene encodes a protein with a predicted mr of 104k which does not share homology with any previously reported proteins. a possible promoter is located four nucleotides upstream of the initiation codon and represents a consensus baculovirus late promoter (ataag). this has been confirmed by the identif ...19911940861
selection kinetics during serial cell culture passage of mixtures of wild-type autographa californica nuclear polyhedrosis virus and its recombinant ac360-beta-gal.detailed analysis of the selection process in serial co-infections of cell cultures by wild-type autographa californica nuclear polyhedrosis virus (acnpv) strain e2 (acnpv/e2) and ac360-beta-gal, a genetically engineered strain, shows that the unaltered strain was clearly dominant even when it initially constituted the minority component in the inoculum. a method of calculating a selection coefficient that quantifies the relative advantage of one strain of virus over the other under specific cul ...19911940862
dna-binding proteins of baculovirus-infected cells during permissive and semipermissive replication.certain insect cell lines have been shown to be permissive (tn-368) or semipermissive (iplb-ld-652y) for autographa californica nuclear polyhedrosis virus (acmnpv) replication (mcclintock et al., 1986b). in this report dna-binding proteins were identified in such cell:virus systems by hybridizing western blots containing uninfected and infected cell proteins with acmnpv or host dna probes. in the acmnpv-infected tn-368 permissive cell system, 8 virus-induced dna-binding proteins with molecular w ...19911950170
identification of spliced baculovirus rnas expressed late in infection.previous to this study, the autographa californica multicapsid nuclear polyhedrosis virus (acmnpv) was known to express only one spliced rna (spliced ie1 or ie0). we have conducted an analysis of rna expressed during infection of spodoptera frugiperda cells with acmnpv and have identified a set of five additional spliced rnas expressed late in infection. a reverse transcription-polymerase chain reaction analysis was used to confirm the identification of the ls (late, spliced) rnas. s1 nuclease a ...19911962442
heterologous recombination between autographa californica nuclear polyhedrosis virus dna and foreign dna in non-polyhedrin segments of the viral genome.we used the expression vector system of autographa californica nuclear polyhedrosis virus (acnpv) and spodoptera frugiperda insect cells to study mechanisms of recombination in insect cells. we concentrated on the isolation and analysis of heterologous recombinants. the e1 region of human adenovirus type 2 (ad2) was inserted into regions of the acnpv genome which lacked apparent homologies to the polyhedrin region. out of a total of 122 recombinant acnpv plaques, which hybridized to ad2 dna in p ...19911962504
in vitro transactivation of baculovirus early genes by nuclear extracts from autographa californica nuclear polyhedrosis virus-infected spodoptera frugiperda cells.nuclear extracts, prepared from autographa californica nuclear polyhedrosis virus-infected spodoptera frugiperda cells during a time course of infection, were analyzed for activation of early gene transcription and for late gene transcription. the templates used in the in vitro transcription assays contained promoters for baculovirus genes that have been classified as immediate early, delayed early, and late. the promoters were derived from the baculovirus 39k, p26, gp64, and dna polymerase gene ...19921316463
physical and functional interaction of human cytomegalovirus dna polymerase and its accessory protein (icp36) expressed in insect cells.expression of the human cytomegalovirus (hcmv) (ad169) dna polymerase gene under the control of the polyhedrin promoter of autographa californica nuclear polyhedrosis virus in spodoptera frugiperda (sf9) cells has provided a source of highly active cmv dna polymerase. in extracts from cmv-infected cells, the cmv dna polymerase is found strongly associated with an additional polypeptide, icp36. this protein has been identified as the cmv homolog of the herpes simplex virus type 1 ul42 gene produc ...19921318399
expression of feline leukaemia virus gp85 and gag proteins and assembly into virus-like particles using the baculovirus expression vector system.in order to test components of feline leukaemia virus (felv) as subunit vaccines, we have constructed recombinant baculoviruses that express the felv envelope glycoprotein gp85 [autographa californica nuclear polyhedrosis virus (acnpv)-gp85] and the structural protein, gag (acnpvgag). the gag protein is expressed and shed into the medium of infected cells as particles which have a buoyant density on sucrose gradients and appearance by electron microscopy similar to those of authentic felv virion ...19921321215
sequence, genomic organization of the ecori-a fragment of autographa californica nuclear polyhedrosis virus, and identification of a viral-encoded protein resembling the outer capsid protein vp8 of rotavirus.we present the sequence and genomic organization of the ecori-a fragment of the autographa californica multicapsid nuclear polyhedrosis virus, which represents 11% of the acmnpv genome. fifteen putative open reading frames and their respective amino acid sequences are described. one open reading frame is similar to the vp8 protein of rotavirus.19921333113
stimulation of thymidine kinase activity in baculovirus infected cells is not due to a virus-coded enzyme.a polyhedrin-positive recombinant autographa californica nuclear polyhedrosis virus (acnpv) carrying a herpes simplex virus thymidine kinase gene under the control of the synxiv promoter, a fusion of synthetic and linker-modified polyhedrin promoters, has been constructed. when this recombinant baculovirus was used to infect a variant of spodoptera frugiperda cells deficient in thymidine kinase (tk-), a high level of tk activity was detected. these results, in conjunction with the demonstration ...19921333755
replication inhibition by nucleoside analogues of a recombinant autographa californica multicapsid nuclear polyhedrosis virus harboring the herpes thymidine kinase gene driven by the ie-1(0) promoter: a new way to select recombinant baculoviruses.the expression of the thymidine-thymidylate kinase (hsv1-tk), (atp: thymidine 5'-phosphotransferase; ec 2.7.1.21) of herpes simplex virus type 1 endows the host cell with a conditional lethal phenotype which depends on the presence of nucleoside analogues metabolized by this enzyme into toxic inhibitors of dna replication. to generate a recombinant baculovirus that could be selected against by nucleoside analogs, the hsv1-tk coding sequence was placed under the control of the autographa californ ...19921335569
expression of recombinant acetylcholinesterase in a baculovirus system: kinetic properties of glutamate 199 mutants.the glycophospholipid-linked, amphiphilic form of acetylcholinesterase (ache) from torpedo californica and the hydrophilic form from mouse were overexpressed in sf9 insect cells using the baculovirus expression system. recombinant baculovirus, constructed by inserting ache cdna's into the genome of autographa californica nuclear polyhedrosis virus adjacent to the strong polyhedron promoter, yielded recombinant enzyme varying between 0.5 and 3.8 mg/l. the recombinant enzyme was glycosylated altho ...19921356436
the baculovirus autographa californica nuclear polyhedrosis virus genome includes a papain-like sequence.the published dna sequence that includes the gene for the envelope glycoprotein gp67 of the baculovirus autographa californica nuclear polyhedrosis virus also contains a segment that shows 32% amino acid identity to papain, a cysteine endopeptidase from the papaya plant (carica papaya). the viral papain-like sequence is apparently affected by a frame-shift mutation, but otherwise appears capable of encoding a functional enzyme. catalytically essential amino acids appear to be conserved, as do di ...19921363350
a sensitive model system for in vivo monitoring of baculovirus gene expression in single infected insect cells.we have developed a fast and sensitive system for the in vivo analysis of gene expression in baculovirus infected lepidopteran insect cells. a recombinant baculovirus containing a luciferase gene from the click beetle, pyrophorus plagiophthalamus, under transcriptional regulation of the polyhedrin gene promoter of autographa californica nuclear polyhedrosis virus (acnpv) was used to infect a spodoptera frugiperda cell line. recombinant luciferase could be monitored by luminometry in real-time wi ...19921368236
baculovirus expression of alkaline phosphatase as a reporter gene for evaluation of production, glycosylation and secretion.we have devised a simple and efficient baculovirus expression vector system to evaluate insect tissue culture cells for their capacity to express, glycosylate and secrete foreign proteins. a truncated placental alkaline phosphatase (seap) gene was inserted into the autographa californica nuclear polyhedrosis virus (acmnpv) genome under the transcriptional control of the polyhedrin gene promoter. production levels, glycosylation, and secretion of the recombinant protein were examined in trichoplu ...19921368794
screening of insect cell lines for the production of recombinant proteins and infectious virus in the baculovirus expression system.eight cell lines derived from the insects spodoptera frugiperda, trichoplusia ni, mamestra brassicae, and estigmene acrea were evaluated for recombinant beta-galactosidase and infectious virus production following infection with the baculovirus autographa californica multiple nuclear polyhedrosis virus (acmnpv). production was assessed on a specific (per cell and per microgram of uninfected cellular protein) and on a volumetric (per milliliter) basis. cell density was found to be an important fa ...19921369220
the baculovirus-integrated retrotransposon ted encodes gag and pol proteins that assemble into viruslike particles with reverse transcriptase.ted is a lepidopteran retrotransposon found inserted within the dna genome of the autographa californica nuclear polyhedrosis virus mutant, fp-d. to examine the proteins and functions encoded by this representative of the gypsy family of retrotransposons, the gag- and pol-like open reading frames (orfs 1 and 2) were expressed in homologous lepidopteran cells by using recombinant baculovirus vectors. expression of orf 1 resulted in synthesis of an abundant ted-specific protein (pr55gag) that asse ...19921371168
expression of foreign epitopes on recombinant occlusion bodies of baculoviruses.recombinant polyhedrin proteins of the baculovirus autographa californica nuclear polyhedrosis virus were constructed to serve as immunologic carriers of foreign epitopes. several recombinants containing an influenza haemagglutinin epitope were obtained. three of the five recombinants formed occlusion bodies (obs) and two did not. all of the recombinant polyhedrin proteins reacted with a monoclonal antibody (mab) specific for an influenza epitope in western blots. presentation of the foreign epi ...19921373262
expression of the major core antigen vp7 of african horsesickness virus by a recombinant baculovirus and its use as a group-specific diagnostic reagent.the major core protein, vp7, of african horsesickness virus serotype 4 (ahsv-4), the aetiological agent of a recent outbreak of the disease in southern europe, was expressed in insect cells infected with a recombinant baculovirus containing a cloned copy of the relevant ahsv gene (s7). analyses of its biochemical and antigenic properties confirmed the authenticity of the protein expressed. the high-level expression of vp7 under the control of the strong polyhedrin promoter of autographa californ ...19921378881
presentation of hepatitis b virus pres2 epitope on bluetongue virus core-like particles.a chimeric protein containing most of the hepatitis b virus pres2 region (amino acid residues 1-48) upstream to, and colinear with the amino-terminus of bluetongue virus vp7 protein (pres2-vp7) was expressed by a recombinant autographa californica nuclear polyhedrosis virus (acnpv). the chimeric protein formed btv core-like particles (clps) in spodoptera frugiperda cells only when the cells were coinfected with this recombinant virus and a recombinant baculovirus that expresses unmodified vp7 an ...19921381538
improved efficiency in determining the titer of the autographa californica baculovirus nonoccluded virus.an economical and time-efficient method for titering the autographa californica multiple-embedded nuclear polyhedrosis virus (acmnpv) by the endpoint method is described. the method uses an electronic pipetting device to perform dilutions in the same 60-well microplate as is used for the assay, thus eliminating the need for test tubes or vials for making dilutions. additionally, since small volumes are used for the dilutions, a substantial savings in medium is achieved. the effect of using three ...19921389161
characterization of a baculovirus gene encoding a small conotoxinlike polypeptide.we identified a gene of autographa californica nuclear polyhedrosis virus (acmnpv) that encodes a small cysteine-rich polypeptide which has size and sequence similarity to omega-conotoxins, a class of calcium ion (ca2+) channel inhibitors, found in the venom of cone snails. transcriptional analysis indicated that the 159-bp open reading frame, which we named ctl, and a downstream 984-bp open reading frame are transcribed as a single 1.3-kb bicistronic late rna. the mature ctl gene product was id ...19921404603
purification of pancreatic cholesterol esterase expressed in recombinant baculovirus-infected sf9 cells.a cdna clone encoding the entire coding sequence of rat pancreatic cholesterol esterase (bile salt-stimulated lipase) was subcloned into the baculovirus transfer vector pvl1392 and used to co-transfect spodoptera frugiperda (sf9) insect cells with wild-type autographa californica nuclear polyhedrosis virus (acnpv) dna. two recombinant proteins (m(r) 74 kda and 64 kda) reactive with anti-cholesterol esterase igg were produced and secreted by the infected sf9 cells in large quantities in a time-de ...19921422214
recombinant laminin b1 chains exhibit intact short-arm domains but do not form oligomeric molecules.the human laminin b1 chain has been produced in the baculovirus expression system in sufficient amounts for biochemical and functional studies. a full-length cdna, which was constructed of four partially overlapping clones and verified by in vitro transcription and translation to be functional, was cloned into the transfer vector pvl1392 behind the polyhedrin promoter. the recombinant construct was incorporated by in vivo homologous recombination into the genome of the wild-type baculovirus, aut ...19921425664
expression of cauliflower mosaic virus orf ii in a baculovirus system.the cauliflower mosaic virus orf ii encoding the aphid transmission factor (atf) was mutagenized to introduce a bamhi restriction site upstream from the initiation codon and then cloned into an eukaryotic viral expression vector (autographa californica nuclear polyhedrosis virus). all recombinant viruses tested in spodoptera frugiperda (sf21) cells expressed a protein of about 18 kd which comigrated in page with atf from infected plants. western blotting using an oligopeptide antiserum to atf co ...19921428752
enhancement of polyhedrin nuclear localization during baculovirus infection.polyhedrin is the major component of the nuclear viral occlusions produced during replication of the baculovirus autographa californica multicapsid nuclear polyhedrosis virus (acmnpv). since viral occlusions are responsible for the horizontal transmission of acmnpv in nature, the biosynthesis, localization, and assembly of polyhedrin are important events in the viral replication cycle. we recently defined the sequence requirements for nuclear localization and assembly of polyhedrin. in this stud ...19921433499
baculovirus phosphoprotein pp31 is associated with virogenic stroma.the psti k fragment of autographa californica nuclear polyhedrosis virus (acmnpv) encodes a protein with a molecular weight of 31,000. to define the role of this protein (pp31) in virus infection further, it was overexpressed in bacteria and used to produce polyclonal antiserum. radioimmunoprecipitation analysis indicated that pp31 was synthesized during both the early and late phases of virus infection, consistent with previous analyses indicating that the gene was regulated by tandem early and ...19921433508
promoter influence on baculovirus-mediated gene expression in permissive and nonpermissive insect cell lines.the activities of viral and insect promoters were examined in a range of insect cell lines permissive and nonpermissive for the replication of the baculovirus autographa californica nuclear polyhedrosis virus. recombinant baculoviruses were constructed to place the bacterial chloramphenicol acetyltransferase gene under the control of promoters strongly active in the early, late, or very late stages of virus replication. in fully permissive cells, expression from a very late promoter was 2- to 3- ...19921433522
functional dissection of the autographa californica nuclear polyhedrosis virus immediate-early 1 transcriptional regulatory protein.autographa californica multicapsid nuclear polyhedrosis virus-infected insect cells express a viral immediate-early transcriptional regulatory protein, ie1, that has been shown by transient-expression assays to stimulate the expression of certain baculovirus delayed-early (de) promoters and to inhibit the expression of other immediate-early (ie) genes. it is believed that certain de promoters are activated, in part, by direct interactions between ie1 and enhancer elements located in regions adja ...19921433524
studies on processing, particle formation, and immunogenicity of the hiv-1 gag gene product: a possible component of a hiv vaccine.antigens in a particulate conformation were shown to be highly immunogenic in mammals. for this reason, the particle forming capacity of derivatives of the hiv-1 group specific core antigen p55 gag was assayed and compared dependent on various expression systems: recombinant bacteria, vaccinia- and baculoviruses were established encoding the entire core protein p55 either in its authentic sequence or lacking the myristylation consensus signal. moreover, p55 gag was expressed in combination with ...19921456888
characterization of the nucleotide sequence of the lymantria dispar nuclear polyhedrosis virus dna polymerase gene region.the dna polymerase gene of the lymantria dispar multinucleocapsid nuclear polyhedrosis virus (ldmnpv) was cloned and sequenced. the predicted dna polymerase protein (1113 amino acids, 115.9k) was found to have an amino acid identity of 48% with the corresponding gene of the autographa californica mnpv (acmnpv). it contains five domains associated with substrate binding, primase interaction, and pyrophosphate hydrolysis and three domains associated with 3'-->5' exonuclease activity common to othe ...19921469355
equilibrium and kinetic analysis of autographa californica nuclear polyhedrosis virus attachment to different insect cell lines.the kinetic and equilibrium attachment of autographa californica nuclear polyhedrosis virus (acmnpv) to seven insect cell lines was evaluated. kinetic experiments revealed differences of up to 10-fold in the infection rates among cell lines. equilibrium binding also varied between cell lines and was saturable. the tn 5b1-4 and tn f cell lines had the highest virus binding affinities and infection rates and exhibited diffusion-limited attachment. the rate of infection appears to be limited by the ...19921469356
transient expression assay in a baculovirus system using firefly luciferase gene as a reporter.transient gene expression assays were developed to assess the function of the regulatory sequences of baculoviruses bombyx mori nuclear polyhedrosis virus (bmnpv) and autographa californica nuclear polyhedrosis virus (acnpv) in insect cells of bombyx mori and spodoptera frugiperda, respectively. dna sequences encoding luciferase (luc) of the firefly photinus pyralis was successfully employed in the expression assay as a reporter gene. recombinant plasmids were constructed containing the luc gene ...19921475909
sequence and in vitro translational analysis of a 1629-nucleotide orf in autographa californica nuclear polyhedrosis virus strain e2.the complete nucleotide (nt) sequence of an open reading frame (orf) (map unit 5.1 to 3.8) from autographa californica nuclear polyhedrosis virus strain e2 (acmnpv-e2) has been determined. this 1629-nt orf has a coding potential for a 61-kda pro-rich protein. however, in vitro translation of the 1629-nt orf and sodium dodecyl sulfate-polyacrylamide-gel electrophoresis (sds-page) revealed a 78-kda protein product. the discrepancy between the m(r) predicted by the nt sequence and that obtained fro ...19921487149
site-specific mutagenesis of the 35-kilodalton protein gene encoded by autographa californica nuclear polyhedrosis virus: cell line-specific effects on virus replication.the gene encoding the 35-kda protein (35k gene) located within the ecori-s genome fragment of autographa californica nuclear polyhedrosis virus (acmnpv) is transcribed early in infection. to examine its function(s) with respect to virus multiplication, we introduced specific mutations of this early gene into the acmnpv genome. in spodoptera frugiperda (sf21) culture, deletion of the 35k gene reduced yields of extracellular, budded virus from 200- to 15,000-fold, depending on input multiplicity. ...19921501287
replication of autographa californica baculovirus (acmnpv) in a coleopteran cell line.a coleopteran cell line (age) derived from the cotton boll weevil anthonomus grandis supported replication of autographa californica multiple nuclear polyhedrosis virus (acmnpv). the titer of extracellular virus (ecv) and the number of occlusion bodies (ob) produced in age cells were approximately equal to those produced by a trichoplusia ni cell line (tn-cl1), and the ob produced by both cell lines were equally infectious for t. ni larvae. the identity of the age cell line was established by ch ...19921522049
the autographa californica baculovirus genome: evidence for multiple replication origins.the autographa californica multinucleocapsid nuclear polyhedrosis virus (acmnpv), which is used for the overexpression of eukaryotic genes and is being engineered for possible use as a viral insecticide, has a circular, supercoiled genome of approximately 128 kilobases. despite its widespread use, little is known about the mechanism by which acmnpv replicates. evidence is presented in this report that acmnpv origins of dna replication are repeated sequences each containing several closely relate ...19921529337
nucleotide sequence and transcriptional analysis of the p80 gene of autographa californica nuclear polyhedrosis virus: a homologue of the orgyia pseudotsugata nuclear polyhedrosis virus capsid-associated gene.the 67.2- to 68.5-m.u. region of autographa californica nuclear polyhedrosis virus was sequenced. a large open reading frame (orf) was identified in the clockwise direction on the circular genome map which could potentially encode an 80-kda polypeptide. analysis of the predicted amino acid sequence of this orf indicated that it was a homologue of the p87 capsid-associated gene of orgyia pseudotsugata mnpv with an overall amino acid similarity of 34.3%. a late transcript of 2.1 kb was mapped to t ...19921529529
in vivo transcriptional analysis of three baculovirus genes: evidence of homology between viral and host transcripts.transcripts for the gp64 and polyhedrin genes as well as a 1629 open reading frame (1629 orf) of the autographa californica nuclear polyhedrosis virus were examined in the midgut tissues and hemocytes of uninfected and infected host trichoplusia ni larvae and sf21 cells. polyhedrin-specific transcripts of 1.2, 3.4, and 4.9 kb were expressed in both infected larval tissues and infected sf21 cells. the highest level of expression for polyhedrin-specific transcripts was observed in hemocytes, where ...19921529534
hepatitis delta antigen expressed by recombinant baculoviruses: comparison of biochemical properties and post-translational modifications between the large and small forms.hepatitis delta virus (hdv) encodes only one protein, the hepatitis delta antigen (hdag). two forms of hdag, a large (27 kda) and a small (24 kda) one, participate in the various steps of hdv replication. to further understand the properties of hdag, we have constructed recombinant baculoviruses and expressed both forms of the hdag in insect cells. the gene encoding hdag was placed under the control of the polyhedrin promoter of autographa californica nuclear polyhedrosis virus (acnpv) by homolo ...19921529540
expression of the nucleocapsid protein of dugbe virus and antigenic cross-reactions with other nairoviruses.the small (s) rna segment of dugbe (dug) virus (nairovirus, bunyaviridae) encodes a single protein, the nucleocapsid (n) protein, of m(r) 49.4 kda. cdna derived from the complete coding region for the n protein was cloned into autographa californica nuclear polyhedrosis virus (acnpv) under control of the polyhedrin promoter and used to infect spodoptera frugiperda insect cells. western blotting analysis using monoclonal antibodies demonstrated the production of dug n protein in the infected cell ...19921529645
nucleotide sequence of a transactivating bombyx mori nuclear polyhedrosis virus immediate early gene.the open reading frame (orf) of 1572 bp contained in the 3.8 kb clai fragment of bmnpv encodes a viral regulatory protein which transactivates the delayed early acmnpv 39k gene. transactivation is induced in uninfected cells following transfection with a plasmid containing only the clai fragment. hitherto immediate early gene promoter activity of the included 631 bp leader sequence is evident since no other viral elements are needed for the transcription of the regulatory gene.19921536885
biophysical and biochemical properties of baculovirus-expressed camv p1 protein.cauliflower mosaic virus (camv) gene i encodes a protein (p1) that has been implicated in the control of virus movement in infected plants. to assist in the characterization of the mechanism of action of p1, gene i has been expressed efficiently in spodoptera frugiperda (sf) cells using recombinant baculovirus. control of the expression of camv gene i by the polyhedrin late promoter in the baculovirus autographa californica nuclear polyhedrosis virus (acnpv) resulted in very high levels of p1 ac ...19921549910
expression of the autographa californica nuclear polyhedrosis virus p10 gene: effect of polyhedrin gene expression.two major late proteins, polyhedrin and p10, are synthesized in large quantities in baculovirus infected insect cells. this and the fact that both proteins are dispensable for virus replication, form the basis for the use of these viruses as vector for foreign gene expression. to address the question whether the autographa californica nuclear polyhedrosis virus p10 promotor-driven expression is influenced by the concurrent expression of the polyhedrin gene, several recombinants were constructed ...19921550489
viral and host cellular transcription in autographa californica nuclear polyhedrosis virus-infected gypsy moth cell lines.infection of two gypsy moth cell lines (iplb-ld652y and iplb-ldfb) by the autographa californica multiple-enveloped nuclear polyhedrosis virus (acmnpv) is characterized by extremely attenuated viral protein synthesis followed by a total arrest of all viral and cellular protein production. in this study, acmnpv- and host cell-specific transcription were examined. overall levels of viral rnas in infected gypsy moth cells were, at most measured times, comparable to rna levels from an infected cell ...19921560533
expression and extracellular release of human immunodeficiency virus type 1 gag precursors by recombinant baculovirus-infected cells.the level of synthesis and extracellular release of human immunodeficiency virus type 1 gag by insect cells was analyzed, using eight different recombinants of autographa californica nuclear polyhedrosis virus harboring various constructs of the gag gene, cloned under the polyhedrin promoter. the results obtained suggested that gag expression was mainly regulated at the transcriptional level and was not significantly influenced by posttranslational events, e.g., gag self-assembly, nuclear transp ...19921560545
a structural polypeptide of the baculovirus autographa californica nuclear polyhedrosis virus contains o-linked n-acetylglucosamine.a structural glycopeptide, gp41, derived from the occluded virus of the baculovirus autographa californica nuclear polyhedrosis virus was characterized. the peptide specifically bound wheat germ agglutinin but was not recognized by a panel of seven other lectins. reactivity with wheat germ agglutinin was eliminated by treatment of gp41 with beta-n-acetylglucosaminidase, indicating that n-acetylglucosamine (glcnac) was present as terminal residues. gp41 was efficiently galactosylated by galactosy ...19921583718
differential factor binding at the promoter of early baculovirus gene pe38 during viral infection: gata motif is recognized by an insect protein.regulatory elements interacting with dna-binding proteins have been investigated in the promoter sequence of the early pe38 gene in the autographa californica nuclear polyhedrosis virus (acnpv). a gata motif located 50 nucleotides upstream of the pe38 transcriptional start site is recognized differentially in the course of infection. as demonstrated by footprint and gel mobility shift assays, the gata sequences ttatct are protected by nuclear extracts from uninfected spodoptera frugiperda cells ...19921583720
regulation of delayed-early gene transcription by dual tata boxes.the 39k autographa californica nuclear polyhedrosis virus (acmnpv) gene is highly expressed throughout the virus life cycle and is controlled by tandem promoters that exhibit features of early and late baculovirus promoters. late transcripts initiate at a conserved taag motif, while early transcripts are heterogeneous and initiate near a conserved cagt motif. to define the nucleotide sequences that regulate early transcription of the 39k gene, a series of mutations was generated by substitution ...19921583729
purification and characterization of human salivary-gland prokallikrein from recombinant baculovirus-infected insect cells.a full-length cdna encoding human salivary-gland preprokallikrein was inserted into the baculovirus autographa californica nuclear polyhedrosis virus downstream of the polyhedrin promoter. the gene was expressed in transfected spodoptera frugiperda cells and the recombinant product secreted into the culture medium. by alternating anion-exchange chromatography and gel-filtration steps, twice repeated, prokallikrein was purified to homogeneity, which was confirmed by amino acid analysis and n-term ...19921587272
the anticarsia gemmatalis nuclear polyhedrosis virus polyhedrin gene region: sequence analysis, gene product and structural comparisons.the genomic region of the anticarsia gemmatalis multiple nucleocapsid nuclear polyhedrosis virus (agmnpv) strain 2d encoding the polyhedrin gene was cloned and mapped, and a 2085 bp sphi-psti fragment containing the gene was sequenced. the polyhedrin polypeptide of the parental isolate agmnpv was manually sequenced, and the amino acid sequence obtained agreed with that deduced from the dna coding region sequence. agmnpv and orgyia pseudotsugata mnpv (opmnpv) are similar in terms of promoter stru ...19921588315
localization of juvenile hormone esterase during development in normal and in recombinant baculovirus-infected larvae of the moth trichoplusia ni.the pathogenesis and cellular localization of juvenile hormone esterase (jhe) was examined in larvae of the moth trichoplusia ni, infected with a recombinant baculovirus (autographa californica nuclear polyhedrosis virus: acnpv) engineered to produce high levels of jhe (jhe virus). the course of jhe localization in the recombinant virus infected larvae was compared with that of both wild type acnpv infected, and uninfected larvae, using immunogold electron microscopy. in the jhe virus infected i ...19921589874
secretion of a functional soluble form of neutral endopeptidase-24.11 from a baculovirus-infected insect cell line.neutral endopeptidase (nep; ec 3.4.24.11) is an integral membrane protein found at the plasma membrane of many cell types. a secreted form of nep (sec-nep) was recently obtained by transfection of cos-1 cells with a recombinant expression vector consisting of the cdna encoding the signal peptide of pro-opiomelanocortin fused in-frame to the cdna sequence of the complete ectodomain of rabbit nep [lemay, waksman, roques, crine & boileau (1989) j. biol. chem. 264, 15620-15623]. in order to produce ...19921599410
generation and analysis of defective genomes of autographa californica nuclear polyhedrosis virus.we have generated defective genomes of autographa californica nuclear polyhedrosis virus (acnpv) by serial, undiluted passage in iplb-sf-21 cell culture in an attempt to identify potential cis-acting sequences important for acnpv dna replication. viral dna isolated from some of the 81 serial passages was analyzed by pulsed-field gel electrophoresis, restriction endonuclease analysis, and southern blot hybridization. acnpv-defective genomes appeared to be generated through a series of successivel ...19921602548
analysis of polyhedra morphology mutants of autographa californica nuclear polyhedrosis virus: molecular and ultrastructural features.two new mutants of autographa californica multiple nuclear polyhedrosis virus affected in the morphogenesis of their polyhedra, designated m276 and m934, were investigated. marker transfer experiments demonstrated that the observed phenotype was due exclusively to alterations in the polyhedrin gene. m276 contained a 229 base insertion near the carboxyl terminus coding region which resulted in synthesis of a truncated protein; m934 had a point mutation substituting phenylalanine for leucine at am ...19921607865
nucleotide sequence of the polyhedron envelope protein gene region of the lymantria dispar nuclear polyhedrosis virus.a 6.4 kb region from the lymantria dispar multicapsid nuclear polyhedrosis virus (ldmnpv) genome was sequenced and found to contain open reading frames (orfs) homologous to the polyhedron envelope (pe) protein coding sequence, and the c-terminal half of orf 1, which is a gene located upstream of the pe protein gene in other baculoviruses. the proteins predicted from the ldmnpv genes encoding the pe protein, and orf 1 demonstrated 27 and 34% amino acid sequence identity, respectively, with the co ...19921607868
insecticidal properties of genetically engineered baculoviruses expressing an insect juvenile hormone esterase gene.exploring the possibility of enhancing the properties of baculoviruses as biological control agents of insect pests, we tested the effect of expressing an insect gene (jhe) encoding juvenile hormone esterase. juvenile hormone esterase inactivates juvenile hormone, which regulates the outcome of an insect molt. a cdna encoding the juvenile hormone esterase of heliothis virescens was inserted into the genome of autographa californica nuclear polyhedrosis virus such that the gene was expressed unde ...19921622228
foreign gene expression by a baculovirus vector with an expanded host range.a nuclear polyhedrosis virus (npv) (baculoviridae)-based gene expression system was improved by dna recombination. the bmn cell line established from bombyx mori and the sf21 cell line established from spodoptera frugiperda are non-permissive for autographa californica multicapsid npv (acmnpv) and b. mori npv (bmnpv) replication, respectively. after cotransfection of acmnpv dna and bamhi-digested bmnpv dna into sf21 cells, progeny viruses were isolated by plaque purification on bmn cell monolaye ...19921629707
nucleotide sequence and transcriptional analysis of a gene encoding gp41, a structural glycoprotein of the baculovirus autographa californica nuclear polyhedrosis virus.we have identified and sequenced a region of the autographa californica nuclear polyhedrosis virus (acmnpv) genome encoding the major polyhedron-derived virus structural glycoprotein gp41. the open reading frame is located entirely within the acmnpv sstii-m fragment. the protein sequence does not have hydrophobic regions characteristic of integral membrane proteins and contains a putative o-linked glcnac glycosylation site. gp41 is expressed as a late gene, with transcripts starting within two c ...19921629955
synthesis and toxicity of full-length and truncated bacterial cryivd mosquitocidal proteins expressed in lepidopteran cells using a baculovirus vector.full-length (72k) and truncated (61k) cryivd mosquitocidal proteins of bacillus thuringiensis (bt) were expressed in spodoptera frugiperda cells and larvae of trichoplusia ni using a baculovirus vector to investigate the role of cryivd peptides in toxicity as well as to evaluate further the baculovirus/lepidopteran system for expressing bt proteins. the cryivd genes were inserted into the autographa californica multinucleocapsid nuclear polyhedrosis virus (acmnpv) under control of the polyhedrin ...19921730944
transcription analysis of the ecori d region of the baculovirus autographa californica nuclear polyhedrosis virus identifies an early 4-kilobase rna encoding the essential p143 gene.we have investigated the transcriptional activity of the 60.1- to 68.3-map-unit region of the baculovirus autographa californica nuclear polyhedrosis virus (acmnpv). twelve transcripts mapping to this region were expressed at various times during infection. an early 4.0-kb transcript, potentially coding for a 143-kda peptide essential for viral dna replication, was maximally abundant at 6 h postinfection (p.i.). transcripts of 0.5, 1.1, 1.4, 2.1, and 3.1 kb were most abundant at 12 h p.i., while ...19921731106
the glycoprotein of thogoto virus (a tick-borne orthomyxo-like virus) is related to the baculovirus glycoprotein gp64.thogoto (tho) virus is a tick-transmitted virus which shares morphological and biochemical characteristics with members of the orthomyxoviridae. the genome of thogoto virus comprises six segments of single-stranded, negative sense rna. the complete nucleotide sequence of the fourth largest rna segment of tho virus has been determined from cdna analyses. this rna segment is 1574 nt long and has a coding capacity for a glycoprotein of 512 amino acids with a predicted molecular weight of 57,550 da. ...19921733105
tandemly repeated sequence at the 3' end of the ie-2 gene of the baculovirus orgyia pseudotsugata multicapsid nuclear polyhedrosis virus is an enhancer element.an enhancer element was identified in the virus orgyia pseudotsugata multicapsid nuclear polyhedrosis virus (opmnpv) that is located adjacent to the 3' end of the ie-2 gene and 5' to an open reading frame that codes for a predicted protein that has 37% homology to the acmnpv pe-38 gene. the opmnpv enhancer (ope) consists of a 66-bp element that is tandemly repeated partially or completely 12 times. the ope sequences were shown to increase gene expression from the autographa californica mnpv dela ...19921736547
expression of the mouse c-abl type iv proto-oncogene product in the insect cell baculovirus system.the cellular gene c-abl is the normal homologue of the transforming gene (v-abl) within the genome of the abelson leukaemia virus. the cdna sequence coding for the cellular form of the murine abl gene (c-abl type iv) has been inserted into the baculovirus transfer vector, pac36c, so that the c-abl gene is under the control of the polyhedrin promoter of autographa californica nuclear polyhedrosis virus (acnpv). spodoptera frugiperda cells infected with the recombinant transfer vector in the prese ...19921737071
baculovirus recombinant expressing a secreted form of a transmembrane carcinoma-associated antigen.ga733-2 is a monoclonal antibody-defined, 40-kda glycoprotein antigen that is associated with carcinomas of various origins. hydrophobicity analysis of the protein sequence predicted by complementary dna (cdna) has suggested that the ga733-2 antigen is a type i membrane protein. in this study, the polymerase chain reaction was used in a strategy to omit cdna sequences for the transmembrane and cytoplasmic domains, thereby converting the extracellular domain into a secretory protein. full-length ...19921737342
a structured dynamic model for the baculovirus infection process in insect-cell reactor configurations.a mathematical description of the infection of insect cells with baculovirus in a continuously operated reactor configuration is presented. the reactor configuration consists of one bioreactor in which insect cells (spodoptera frugiperda) are grown followed by one or two bioreactors in which cells are infected by a baculovirus (autographa californica nuclear polyhedrosis virus). it was demonstrated that the so-called passage effect is responsible for the observed difference in run time between a ...199218601149
production of recombinant proteins in high-density insect cell cultures.the effect of the growth phase of spodoptera frugiperda (sf9) cells on the production of recombinant proteins (beta-galactosidase and glucocerebrosidase) was investigated. cells infected with the recombinant autographa californica nuclear polyhedrosis virus at the late exponential and stationary phases yielded low quantities of expressed protein. highest enzyme yields were obtained using sf9 cells from the early exponential phase (0.9 mg beta-galactosidase/10(6) cells and 1.7 microg glucocerebro ...199318612984
effect of temperature and oxygen on cell growth and recombinant protein production in insect cell cultures.the effect of temperature and o2 saturation on the production of recombinant proteins beta-galactosidase and human glucocerebrosidase by spodoptera frugiperda cells (sf9) infected with recombinant autographa californica nuclear polyhedrosis virus was investigated. the rates of cell growth, glucose consumption, o2 consumption and product expression were measured at temperatures between 22 degrees c and 35 degrees c. the results indicated that possible o2 limitation may be alleviated without compr ...19937763472
host-specific transcription of baculovirus genes.we examined the differential replication of autographa californica multicapsid nuclear polyhedrosis virus (acmnpv; family: baculoviridae) in spodoptera frugiperda (sf) and bombyx mori (bm) cell lines. our previous studies have shown that infection of bm cells is abortive; infectious progeny is not produced, and although virogenic stroma is produced, complete virions or inclusion bodies are not assembled. we designed the present study to determine the extent of infected-cell-specific polypeptides ...19937763702
production of human alkaline phosphatase, a secreted, glycosylated protein, from a baculovirus expression system and the attachment-dependent cell line trichoplusia ni bti-tn 5b1-4 using a split-flow, air-lift bioreactor.a split-flow, air-lift bioreactor for the cultivation of insect cells to produce recombinant protein is described. it can be used advantageously with attached cell systems. this bioreactor incorporates two sections: a rise and a downcomer. trichoplusia ni bti-tn 5b1-4 cells are grown on a support material of glass beads or microcarriers placed in the downcomer. this cell line is more productive than other commonly used insect cell lines, but it has the disadvantage of being difficult to use at l ...19937764358
glycosylation and high-level secretion of human tumour necrosis factor-beta in recombinant baculovirus-infected insect cells.human tumour necrosis factor-beta (tnf-beta) was produced in eukaryotic cells using the insect baculovirus cloning and expression system. a novel insect signal sequence, the honey-bee (apis mellifera) prepromelittin secretory sequence, was used to aid in the post-translational modifications, glycosylation and secretion of recombinant human tnf-beta. human tnf-beta cdna was cloned using the insect baculovirus vector pacc4s. expression of the human tnf-beta was regulated by the insect autographa c ...19938297505
the 1629-bp open reading frame of the autographa californica multinucleocapsid nuclear polyhedrosis virus encodes a virion structural protein.a 1629-bp open reading frame (orf) of autographa californica multinucleocapsid nuclear polyhedrosis virus (acmnpv) is shown to encode a 78-kda virion structural protein. to determine this, polyclonal antibody was made to a fusion protein synthesized in escherichia coli from a chimeric gene that contained 1415 bp of the 1629-bp gene. in western blot analyses, this antibody cross-reacted with a protein of about 78 kda in both extracellular virions (ecv) and virions isolated from polyhedra (pdv), a ...19938299959
identification and immunocharacterization of a 25k structural protein in autographa californica nuclear polyhedrosis virus (acmnpv).a 25k protein was a major component of the triton x-100 solubilized fraction prepared for the isolation of nuclear matrices from spodoptera frugiperda cells infected with autographa californica nuclear polyhedrosis virus (acmnpv). polyclonal antisera to p25 were produced in rabbits and used in the characterization of p25. western blots of proteins probed with the p25 antiserum demonstrated the presence of the p25 in both the polyhedral-derived and extracellular viral phenotypes. using an indirec ...19938317143
non-homologous recombination between adenovirus and acnpv dna fragments in cell-free extracts from insect spodoptera frugiperda nuclei.in previous work, we have developed a cell-free system from nuclear extracts of hamster cells to study the mechanism of integrative recombination between adenovirus type 12 (ad12) dna and hamster cell dna (jessberger et al., 1989; tatzelt et al., 1992). we have also demonstrated that in insect cells the left terminal fragment of ad2 dna can insert by non-homologous recombination into the 32.6 to 34.5 map unit (ecori-o) fragment and into other segments of autographa californica nuclear polyhedros ...19938317144
one-step purification and characterization of human pancreatic lipase expressed in insect cells.a cdna clone encoding the sequence of human pancreatic lipase (hpl) was subcloned into the baculovirus transfer vector pvl1392 and used in co-transfection of spodoptera frugiperda (sf9) insect cells with wild-type autographa californica nuclear polyhedrosis virus (acnpv) dna. a single recombinant protein (50 kda) secreted by sf9 cells was detectable in the culture medium 24 h post-infection using both anti-hpl polyclonal antibodies and potentiometric measurements of lipolytic activity. the expre ...19938335100
a 25-kda protein is associated with the envelopes of occluded baculovirus virions.antiserum produced against preoccluded virions of the orgyia pseudotsugata multinucleocapsid nuclear polyhedrosis virus (opmnpv) was used to screen an opmnpv lambda gt11 expression library. one of the immunoreactive clones contained an insert that hybridized to a portion of the opmnpv hindiii-p fragment. a 2-kb region of hindiii-p was sequenced and found to contain three open reading frames, each preceded by a late promoter element. the insert from the lambda gt11 clone was derived from the thir ...19938337828
transcription mapping and functional analysis of the protein tyrosine/serine phosphatase (ptpase) gene of the autographa californica nuclear polyhedrosis virus.the protein tyrosine/serine phosphatase (ptpase) gene of the autographa californica nuclear polyhedrosis virus yields two major transcripts of approximate sizes 3.1 and 3.9 kb. both of these are very late transcripts, which accumulate to maximal levels more than 30 hr postinfection. the smaller transcript initiates at the t of an ataag sequence that lies 22 bp upstream of the putative start codon of the gene. the larger transcript initiates at the first a of a ttaag sequence that lies approximat ...19938337833
immediate-early baculovirus genes transactivate the p143 gene promoter of autographa californica nuclear polyhedrosis virus.trans-acting regulatory components of autographa californica nuclear polyhedrosis virus (acmnpv) were studied in a transient assay system for their ability to activate the p143 gene promoter. a region including 502 nt upstream of the aug of the p143 gene was linked to the bacterial chloramphenicol acetyltransferase (cat) gene. the p143 promoter-cat construct was used to identify acmnpv immediate-early gene products required for expression from the p143 gene early promoter. transient expression a ...19938337840
characterization of an early gene coding for a highly basic 8.9k protein from the orgyia pseudotsugata multicapsid nuclear polyhedrosis virus.a new gene of the baculovirus orgyia pseudotsugata multicapsid nuclear polyhedrosis virus (opmnpv) has been identified that encodes a highly basic 8.9k protein. the gene called p8.9 is expressed as a 0.5 kb transcript by 1 h post-infection and initiates at an early gene motif. the promoter of the 0.5 kb transcript has two upstream elements, repeats i and ii, which are similar to motifs previously characterized in the opmnpv ie-2 gene and the autographa californica nuclear polyhedrosis virus ie-n ...19938345350
expression of human thyroid peroxidase in insect cells using recombinant baculovirus.the baculovirus expression system was used to overexpress recombinant human thyroid peroxidase. sf-9 cells infected with the recombinant virus acmnpv-htpo synthesized htpo protein (htpo-bac) immunogenic on western blots when probed with either rabbit anti-tpo peptide sera or pooled human anti-tpo sera (ms12/89). htpo-bac was a major constituent of the membrane fraction from the infected cells, constituting 14.9% and 10.1% of the 1% deoxycholate-soluble and insoluble fractions, respectively, as j ...19938349029
identification, sequence, and transcriptional mapping of lef-3, a baculovirus gene involved in late and very late gene expression.a trans-acting gene required for late viral gene expression in transient expression assays was identified in the genome of autographa californica nuclear polyhedrosis virus. a genomic library of a. californica nuclear polyhedrosis virus dna lacking a clone spanning the region from 43 to 48 map units (mu) was unable to activate gene expression from a reporter plasmid when the reporter gene was driven by a baculovirus late or very late promoter in transient expression assays. the genomic region re ...19938350397
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