Publications

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 ...	1991	1924311
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 ...	1991	1926775
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 ...	1991	1937047
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 ...	1991	1940861
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 ...	1991	1940862
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 ...	1991	1950170
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 ...	1991	1962442
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 ...	1991	1962504
purification and refolding of recombinant human igf ii from silkworms infected with recombinant bombyx mori nuclear polyhedrosis virus.	we have reported that insulin-like growth factor ii (igf ii) was produced as a fusion protein in bombyx mori (silkworm) larval bodies infected with recombinant b. mori nuclear polyhedrosis virus [j. gen. virol., 68, 2599-2606 (1987)]. in this study, the purification of igf ii from the infected silkworms is reported. the fusion protein was extracted with 6.0 m guanidine-hcl from the infected larval bodies homogenized in water. the use of organic solvents to remove the impurities, such as lipid de ...	1992	1368125
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 ...	1992	1368236
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 ...	1992	1368794
high level expression of a frog alpha-amidating enzyme, ae-ii, in cultured cells and silkworm larvae using a bombyx mori nuclear polyhedrosis virus expression vector.	a xenopus laevis peptidyl c-terminal alpha-amidating enzyme (ae-ii) gene, modified by deletion of a region encoding the putative membrane-spanning domain and the putative c-terminal cytosolic tail, was expressed in bomo-15 aiic insect cells and silkworm larvae using a bombyx mori baculovirus expression vector system. the expressed enzyme was identified predominantly in the culture medium and the hemolymph of silkworm larvae, indicating successful secretion of the expressed ae-ii. the level of re ...	1992	1368810
antibody production in silkworm cells and silkworm larvae infected with a dual recombinant bombyx mori nuclear polyhedrosis virus.	we have examined the efficiency of coexpression of two heterologous genes from a recombinant bombyx mori nuclear polyhedrosis virus for the production of antibodies in silkworm larvae. the cdnas encoding the light and the heavy chains of a murine immunoglobulin, directed against lipoprotein i of pseudomonas aeruginosa, were brought under the control of two separate copies of the viral polyhedrin promotor. infection of silkworm cells with the recombinant baculovirus yielded a maximum of 6.4 micro ...	1992	1368987
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 ...	1992	1369220
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 ...	1992	1371168
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 ...	1992	1373262
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 ...	1992	1378881
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 ...	1992	1381538
potential role of monocyte chemoattractant protein 1/je in monocyte/macrophage-dependent iga immune complex alveolitis in the rat.	we have examined the role of monocyte chemoattractant protein 1 (mcp 1) in the pathogenesis of monocyte/macrophage-dependent iga immune complex alveolitis in the rat. rat mcp 1 was cloned and expressed in order to facilitate analysis of its function in rat models of human disease. a cdna library was constructed from rat pulmonary artery endothelial cells stimulated with tnf-alpha. the cdna library was screened with synthetic oligonucleotide probes based on the recently published rat mcp 1 cdna s ...	1992	1387671
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 ...	1992	1389161
biochemical characterization and time-course analysis of lymantria dispar nuclear polyhedrosis virus with monoclonal antibodies.	hybridoma cell lines secreting monoclonal antibodies (mabs) specific to a 31,000 molecular weight viral protein or a 31,000 molecular weight polyhedrin protein of lymantria dispar nuclear polyhedrosis virus (ldnpv) were developed. the two polypeptides were shown to be different by comparing their amino acid compositions. immuno-electron microscopy was used to verify specific binding of the mabs to their respective targets. specific mabs were used to develop an elisa procedure to monitor the deve ...	1992	1393827
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 ...	1992	1404603
baculovirus gp64 envelope glycoprotein is sufficient to mediate ph-dependent membrane fusion.	the baculovirus gp64 envelope glycoprotein is a major component of the envelope of the budded virus (bv) and is involved in bv entry into the host cell by endocytosis. to determine whether gp64 alone was sufficient to mediate membrane fusion, the orgyia pseudotsugata multicapsid nuclear polyhedrosis virus gp64 protein was transiently expressed in uninfected insect cells. cells expressing the baculovirus gp64 protein were examined for membrane fusion activity by using a syncytium formation assay ...	1992	1404622
hyperexpression of baculovirus polyhedrin and p10 is inversely correlated with actin synthesis.	polyhedrin and p10, two proteins encoded by autographa californica m nuclear polyhedrosis virus, are hyperexpressed very late during normal infections. in this study we found that cytochalasin d, a drug that leads to increased actin synthesis in infected and uninfected host cells, delayed the amplified expression of polyhedrin and p10 when added to infected cells before hyperexpression was already in progress. restoration of polyhedrin and p10 hyperexpression could be achieved by removal of the ...	1992	1413514
identification of bent dna and ars fragments in the genome of choristoneura fumiferana nuclear polyhedrosis virus.	we identified four cfmnpv dna fragments with autonomously replicating sequences (ars) functional in saccharomyces cerevisiae. a 0.9-kb fragment which, mapped to 54.5 to 55.3 map units within ecori hi of the cfmnpv genome, showed the strongest ars activity of the four. sequence analysis of this 0.9-kb dna segment revealed an a+t-rich region separated from a g+c-rich region by 320 bp. although no sequence matched exactly the ars core-consensus sequence, 13 near-matches differing by only one or two ...	1992	1413988
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 ...	1992	1422214
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 ...	1992	1425664
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 ...	1992	1428752
nucleotide sequence and transcriptional analysis of the polyhedrin gene of spodoptera exigua nuclear polyhedrosis virus.	the nucleotide sequence of a 1.1 kbp fragment of the multiple nucleocapsid nuclear polyhedrosis virus (mnpv) of spodoptera exigua (se) containing the polyhedrin gene was determined. an open reading frame (orf) of 738 nucleotides (nt) was detected. this orf encoded a protein of 246 amino acids with a predicted m(r) of 29k. the nucleotide and amino acid sequences were compared with the sequences of eight other npv polyhedrins. the semnpv polyhedrin protein was most closely related to s. frugiperda ...	1992	1431809
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 ...	1992	1433499
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 ...	1992	1433508
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- ...	1992	1433522
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 ...	1992	1433524
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 ...	1992	1469355
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 ...	1992	1469356
protein synthesis in pupae of the silkworm bombyx mori after infection with nuclear polyhedrosis virus: resistance to viral infection acquired during pupal period.	protein synthesis has been studied in pupae of the silkworm bombyx mori (bm) infected with nuclear polyhedrosis virus (bmnpv) at various stages of the pupal period. nascent proteins were labelled by injection of [35s]methionine into pupae and then analysed by sds-page. temporal regulation of synthesis of infected cell-specific proteins (icsps) in pupae was demonstrated by electrophoretic analysis of the proteins labelled at different times post-infection (p.i.). the rate of icsp synthesis reache ...	1992	1469357
induction of a novel protein kinase in pupae of the silkworm bombyx mori after infection with nuclear polyhedrosis virus.	protein kinases induced by bombyx mori nuclear polyhedrosis virus in pupae of the silkworm b. mori were examined by activity gel analysis using phosvitin as a protein substrate. the method involved page of the soluble fraction from pupae under native conditions and in the presence of sds, followed by in situ renaturation of proteins and recovery of protein kinase activity in the intact gel. a novel protein kinase able to phosphorylate phosvitin was detected in the infected pupae from 2 days post ...	1992	1469362
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 ...	1992	1475909
characterization of baculovirus-expressed hemagglutinin and fusion glycoproteins of the attenuated measles virus strain aik-c.	with measles virus cdna of the avirulent vaccine strain aik-c, two cdnas of h or f genes were amplified by the polymerase chain reaction. the amplified cdnas were inserted respectively to the baculovirus transfer vector pacym1 derived from the nuclear polyhedrosis virus of autographa californica (acnpv). after co-transfection of the transfer vectors with acnpv dna to spodoptera frugiperda cells, recombinant baculoviruses were screened by plaque assay, and the viruses containing h-cdna or f-cdna ...	1992	1476005
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 ...	1992	1487149
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. ...	1992	1501287
a synthetic early promoter from a baculovirus: roles of the tata box and conserved start site cagt sequence in basal levels of transcription.	many baculovirus early genes and insect genes transcribed by rna polymerase ii have a conserved transcription start site sequence (cagt) located downstream of a consensus tata box. to examine the functions and interactions of these two motifs in initiating accurately positioned basal transcription, a 43-nt synthetic promoter was synthesized from the tata box and start site sequences of the gp64 early promoter from the orgyia pseudotsugata multicapsid nuclear polyhedrosis virus (opmnpv). the synt ...	1992	1519359
a cellular promoter-based expression cassette for generating recombinant baculoviruses directing rapid expression of passenger genes in infected insects.	we have developed an expression cassette which allows the generation of recombinant baculoviruses that can express passenger genes under the control of a constitutive cellular promoter derived from the cytoplasmic actin gene of the silkmoth bombyx mori. silkmoth tissue culture cells which were infected with a recombinant b. mori nuclear polyhedrosis virus (bmnpv) containing the gene-encoding chloramphenicol acetyltransferase (cat) under the control of this expression cassette expressed significa ...	1992	1519361
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 ...	1992	1522049
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 ...	1992	1529337
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 ...	1992	1529529
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 ...	1992	1529534
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 ...	1992	1529540
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 ...	1992	1529645
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.	1992	1536885
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 ...	1992	1549910
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 ...	1992	1550489
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 ...	1992	1560533
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 ...	1992	1560545
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 ...	1992	1583718
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 ...	1992	1583720
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 ...	1992	1583729
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 ...	1992	1587272
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 ...	1992	1588315
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 ...	1992	1589874
a simple method for the extraction of baculovirus dna.	we have developed a method using lysozyme for dna extraction from baculoviruses using as a model lymantria dispar nuclear polyhedrosis virus (ldnpv) obtained from infected larvae. this method proved to be quick, inexpensive and the extracted dna was successfully used in molecular hybridization experiments.	1992	1597509
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 ...	1992	1599410
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 ...	1992	1602548
the amino acid sequence determination of a granulin and polyhedrin from two baculoviruses infecting agrotis segetum.	the amino acid sequence of agrotis segetum granulosis virus (asgv) granulin and a. segetum nuclear polyhedrosis virus (asnpv) polyhedrin was determined by sequencing tryptic and chymotryptic peptides from reduced and carboxymethylated proteins and tryptic fragments of oxidized and maleylated granulin. the comparison of the established peptide structures with the primary structures of other occlusion body proteins from related baculoviruses was also used for the polypeptide chains' reconstruction ...	1992	1604817
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 ...	1992	1607865
dissimilar expression of autographa californica multiple nucleocapsid nuclear polyhedrosis virus polyhedrin and p10 genes.	the temporal expression of the autographa californica multiple nucleocapsid nuclear polyhedrosis virus polyhedrin and p10 genes in spodoptera frugiperda cells was studied using virus recombinants in which either gene was replaced by the juvenile hormone esterase (jhe) gene of heliothis virescens. the jhe served as a highly specific and sensitive reporter for gene expression. activation of the p10 gene followed a pattern different to that of polyhedrin. the p10 gene was activated a few hours earl ...	1992	1607866
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 ...	1992	1607868
nucleotide sequence of the p39-capsid gene region of the lymantria dispar nuclear polyhedrosis virus.	a 1.85 kb region, containing an open reading frame (orf) homologous to the baculovirus p39-capsid gene, was sequenced from the lymantria dispar multicapsid nuclear polyhedrosis virus (ldmnpv) genome. analysis of the p39-capsid gene demonstrated that it was 39% and 47% identical in amino acid sequence with the homologous genes in the autographa californica and orgyia pseudotsugata mnpvs, respectively. two late promoter elements located upstream of the p39 gene in the ldmnpv genome are conserved w ...	1992	1607869
genetic engineering of a lymantria dispar nuclear polyhedrosis virus for expression of foreign genes.	a bacterial lacz gene was inserted into an isolate of the lymantria dispar nuclear polyhedrosis virus (ldmnpv). the transfer vector was constructed by site-directed mutagenesis of the translation start site of the ldmnpv polyhedrin gene, within the bglii e fragment of the viral genome. a multiple cloning sequence was inserted at this start site and used for the insertion of the lacz gene into the transfer plasmid. liposome transfection was used to cotransfect l. dispar tissue culture cells with ...	1992	1607870
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 ...	1992	1622228
binding of low concentration of peptide to h-2kd produced in insect cells requires mouse beta 2-microglobulin co-expression.	a recombinant baculovirus expressing the murine class i mhc heavy chain h-2kd cdna under the transcriptional control of autografa californica nuclear polyhedrosis virus (acnpv) polyhedrin promoter has been isolated and used to infect sf9 lepidopteran cells either alone or in association with a previously isolated virus expressing mouse beta 2-microglobulina (beta 2-ma). when infected with the heavy chain-encoding virus alone, h-2kd was produced in a beta 2-m-free conformation detected on the sur ...	1992	1622899
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 ...	1992	1629707
characterization of two variants of panolis flammea multiple nucleocapsid nuclear polyhedrosis virus.	the baculovirus panolis flammea multiple nucleocapsid nuclear polyhedrosis virus (pfmnpv) was originally isolated from a natural virus epizootic and shown to consist of a mixture of variants. two subclasses of variants (pfmnpv a and b) were identified by southern blot hybridization, their polyhedrin genes being located on different restriction fragments. the proportion of the a and b variants changed according to the larval host in which the virus was propagated; pfmnpv(b) predominated in p. fla ...	1992	1629708
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 ...	1992	1629955
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 ...	1992	1730944
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 ...	1992	1731106
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. ...	1992	1733105
molecular analysis of the trans-activating ie-2 gene of orgyia pseudotsugata multicapsid nuclear polyhedrosis virus.	a second immediate early (ie) regulatory gene of the baculovirus orgyia pseudotsugata multicapsid nuclear polyhedrosis virus (opmnpv) has been identified. the ie-2 gene which is homologous to the ie-n gene of autographa californica mnpv was mapped to the hindiii a fragment of opmnpv between 0.41 to 1.37 map units. the ie-2 gene codes for a predicted protein of 45,640 da and analysis of the amino acid sequence shows that the protein has a highly basic amino terminal domain and a cysteine-rich dom ...	1992	1736546
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 ...	1992	1736547
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 ...	1992	1737071
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 ...	1992	1737342
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 ...	1992	1316463
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 ...	1992	1318399
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 ...	1992	1321215
expression in insect cells and immune reactivity of a 28k tegument protein of human cytomegalovirus.	the gene encoding the highly antigenic 28k (pp28) tegument phosphoprotein of human cytomegalovirus (hcmv) was expressed in insect cells utilizing a recombinant baculovirus. the mature intracellular form of the recombinant-derived pp28 had mobility on sds-polyacrylamide gels similar to that of native pp28 from hcmv strain towne-infected human foreskin fibroblasts (hffs). in vitro labelling of recombinant autographa california nuclear polyhedrosis virus-infected spodoptera frugiperda cells or of h ...	1992	1328492
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.	1992	1333113
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 ...	1992	1333755
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 ...	1992	1335569
expression and characterization of kinase-active v-erbb protein using a baculovirus vector system.	the v-erbb gene is an oncogene of the avian erythroblastosis virus encoding a protein that is a truncated version of the epidermal growth factor receptor. the v-erbb protein was expressed alone or as polyhedrin-erbb fusion proteins using the bombyx mori nuclear polyhedrosis virus vector. the expression level of the fusion protein whose polyhedrin portion consisted of only 8 amino-terminal amino acids was more than ten times higher than that of the non-fusion protein. studies with tunicamycin sho ...	1992	1347525
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 ...	1992	1356436
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 ...	1992	1363350
continuous beta-galactosidase production with a recombinant baculovirus insect-cell system in bioreactors.	insect cells were exploited to produce bacterial beta-galactosidase by infecting them with a recombinant nuclear polyhedrosis virus (baculovirus) of autographa californica. the insect cells were cultured in a continuous stirred tank reactor (cstr) and led to a second cstr where they were infected with a recombinant virus in which the lacz gene from escherichia coli was inserted. in the effluent of the production reactor, maximum activities of 15 units beta-galactosidase per 10(6) cells were meas ...	1992	1367985
production of a discrete, heterogeneous population of beta-galactosidase polypeptides using baculovirus expression vectors.	gel electrophoresis analysis of immunoprecipitated beta-galactosidase and polyhedrin-beta-galactosidase expressed in spodoptera frugiperda cells infected with recombinant autograph californica nuclear polyhedrosis virus revealed the existence of a population of discrete beta-galactosidase polypeptides. several of the polypeptides observed in the fusion protein expression experiments exhibit a consistent pattern of slightly greater molecular weight when compared to the nonfusion beta-galactosidas ...	1992	18601031
investigation of reduced serum and serum-free media for the cultivation of insect cells (bm5) and the production of baculovirus (bmnpv).	an experimental study has been carried out to investigate the effectivenes of several reduced serum and serum-free media for the cultivation of an ovarian cell line, bm5, of the lepidopteran insect bombyx mori. bm5 cell were successfully adapted to grow in a medium containing 5% serum and a serum-free medium (ex-cell 400). on the other hand, this cell line could not be adapted to grow in several other media suggested in the literature, including ipl-41 + 2% fetal bovine serum (fbs), sf-900, and ...	1992	18601067
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 ...	1992	18601149
transfer vectors for maximal expression of passenger genes in the bombyx mori nuclear polyhedrosis virus expression system.	a series of bombyx mori nuclear polyhedrosis virus (bm-npv) transfer vectors has been developed containing various lengths of the polyhedrin promoter, including sequences 3' of the initiation codon. the atg initiation codon was mutated in some of these vectors to allow for the production of authentic nonfusion proteins. the ability of the various polyhedrin promoter constructs to direct expression of foreign gene sequences was assessed using two test genes, chloramphenicol acetyl transferase (ca ...	1993	18612957
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 ...	1993	18612984
a two-stage bioreactor system for the production of recombinant proteins using a genetically engineered baculovirus/insect cell system.	a two-stage bioreactor scheme was developed for the large-scale production of recombinant proteins using a genetically engineered baculovirus/insect cell system. the first bioreactor was employed for cell growth and the second for cell infection. silkworm bm5 cells were infected with a recombinant baculovirus, bmnpv/p5.cat, containing a bacterial chloramphenicol acetyltransferase (cat) gene under the control of the polyhedrin gene promoter of bombyx mori nuclear polyhedrosis virus (bmnpv). this ...	1993	18613020
baculovirus gene me53, which contains a putative zinc finger motif, is one of the major early-transcribed genes.	we have been investigating the first viral genes to be transcribed in spodoptera frugiperda cells after the infection with autographa california nuclear polyhedrosis virus in order to identify regulatory proteins required for the activation of early and/or late gene transcription. by screening a cdna library from polyadenylated rna transcribed at 1 h postinfection, we identified a gene which is designated me53. this newly identified gene is located upstream of the late p74 gene and forms a diver ...	1993	8093490
physical mapping of buzura suppressaria nuclear polyhedrosis virus genome.	a physical map of the genome of buzura suppressaria singly-enveloped nuclear polyhedrosis virus (bssnpv) was determined using six restriction endonucleases (re), singly or in pairs, and southern hybridizations of cloned or gel-purified dna fragments. the maps of the genomes of bssnpv and autographa californica multiply-enveloped nuclear polyhedrosis virus (acmnpv) were different.	1993	8094612