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nucleotide sequencing and transcriptional mapping of the orgyia pseudotsugata multicapsid nuclear polyhedrosis virus p10 gene.a 32p-labeled cloned dna fragment (acmnpv hindiii-q) containing one of the repeated sequences and a portion of the p10 gene from autographa californica multicapsid nuclear polyhedrosis virus (acmnpv) was used to probe southern blots containing restriction endonuclease digests of orgyia pseudotsugata muticapsid nuclear polyhedrosis virus (opmnpv) dna. a single 3.6-kb fragment, opmnpv hindiii-q, was hybridized. the opmnpv hindiii-q fragment was cloned into puc-18, mapped with restriction endonucle ...19863526709
identification of an amino acid essential to the normal assembly of autographa californica nuclear polyhedrosis virus polyhedra.we compared the dna sequence of the autographa californica nuclear polyhedrosis virus polyhedrin gene with that of the polyhedrin gene from a morphology mutant called m5. a single point mutation was found at the bamhi restriction site within the polyhedrin coding sequence. this point mutation caused a substitution of leucine for proline at amino acid 58 in the m5 polyhedrin. this point mutation was shown to be responsible for both the appearance of cubic polyhedra and the altered mobility of the ...19863528527
cell-surface expression of influenza virus haemagglutinin in insect cells using a baculovirus vector.a baculovirus transfer vector was constructed using a cloned copy of the autographa californica nuclear polyhedrosis virus (acnpv) polyhedrin gene. the atg translation initiation codon and 32 bases of the polyhedrin mrna 5' leader sequences were deleted and replaced with a cdna clone of the a/pr/8/34 influenza virus haemagglutinin (ha) gene. the hybrid ha/polyhedrin gene was transferred to acnpv and polyhedrin-negative recombinants identified which expressed ha activity. the insect cell-derived ...19863529699
physical mapping of temporally regulated, overlapping transcripts in the region of the 10k protein gene in autographa californica nuclear polyhedrosis virus.we investigated transcriptional activity in the region of a gene for a major late protein (10 kilodaltons) of autographa californica nuclear polyhedrosis virus. this 10k protein gene spans an hindiii cleavage site, with the 5' end of the gene located in the hindiii-q fragment and the 3' end in the hindiii-p fragment. northern blot analysis showed that there were at least four transcripts mapping in this region, two that are present maximally at 12 h (1,500 and 1,100 bases in length) and two that ...19862867228
functional mapping of a trans-activating gene required for expression of a baculovirus delayed-early gene.the temporal regulation of an early gene of the baculovirus autographa californica nuclear polyhedrosis virus was examined. we constructed a plasmid (plasmid 39cat) in which the bacterial gene for chloramphenicol acetyltransferase was placed under the control of the promoter for the gene for a a. californica nuclear polyhedrosis virus 39,000-dalton protein (39k). a transient expression assay of plasmid 39cat revealed that the 39k gene was expressed in infected cells but not in uninfected cells, ...19863944847
autographa californica nuclear polyhedrosis virus efficiently enters but does not replicate in poikilothermic vertebrate cells.the host range of the insect virus autographa californica nuclear polyhedrosis virus (acmnpv) was examined. acmnpv could not initiate a productive infection in frog, turtle, trout, or codling moth cell lines. after exposure to acmnpv, neither viral dna nor rna synthesis could be detected in these cell lines as assayed by nucleic acid probe hybridization. entry of acmnpv nucleocapsids into the cytoplasm and viral dna into the nucleus, however, was as efficient in the nonpermissive cell lines as i ...19863301733
the 64k envelope protein of budded autographa californica nuclear polyhedrosis virus. 19863545692
expression of the autographa californica nuclear polyhedrosis virus genome in insect cells: homologous viral and heterologous vertebrate genes--the baculovirus vector system. 19863545694
baculovirus expression vectors: the requirements for high level expression of proteins, including glycoproteins.the requirements for high level expression of three foreign proteins using the polyhedrin gene promoter of autographa californica nuclear polyhedrosis virus (acnpv, baculoviridae) have been investigated. in spodoptera frugiperda cells infected with the appropriate recombinant baculoviruses, the synthesis of the two s rna coded genes of lymphocytic choriomeningitis virus (lcmv; i.e. the nucleoprotein, n, and glycoprotein precursor, gpc), or the haemagglutinin gene of influenza a virus, appears to ...19873553425
expression of the human immunodeficiency virus gag gene in insect cells.regions of the gag-pol gene of human immunodeficiency virus (hiv), the causative agent of aids, have been cloned into the polyhedrin gene of the baculovirus autographa californica nuclear polyhedrosis virus. when these recombinant viruses were used to infect insect cells, the cells produced gag-related proteins which could be immunoprecipitated with serum from aids patients. the major proteins produced by acgag1, which contained the entire gag gene and a small portion of the pol gene, had molecu ...19873554734
comparison of the p26 gene region of two baculoviruses.a 1.1-kb region of dna containing the p26 gene of the multicapsid nuclear polyhedrosis virus of orgyia pseudotsugata (opmnpv) was sequenced, transcriptionally mapped, and compared to the same region in the mnpv of autographa californica (acmnpv). the mrna start site of the p26 gene occurs about 22 nucleotides downstream from an a/t-rich putative promoter sequence that is highly conserved between acmnpv and opmnpv. the p26 mrna is transcribed through the p26 gene and coterminates with the p10 gen ...19873686831
secretion of particles of hepatitis b surface antigen from insect cells using a baculovirus vector.the coding sequences of the hepatitis b virus surface antigen were inserted into a baculovirus transfer vector produced from autographa californica nuclear polyhedrosis virus (acnpv) so that the foreign gene was under the control of the acnpv polyhedrin promoter. spodoptera frugiperda cells infected with the derived recombinant baculovirus produced and secreted 22 nm particles containing the hepatitis b surface antigen. the particles had morphological and antigenic properties identical to those ...19873312490
expression of envelope glycoproteins of human immunodeficiency virus by an insect virus vector.the envelope gene of human immunodeficiency virus was inserted into the genome of an insect virus vector (autographa californica nuclear polyhedrosis virus). upon infection of tissue culture cells, this recombinant virus produced immunoreactive polypeptides related to the envelope glycoproteins of human immunodeficiency virus. serological survey indicates such polypeptides would be of value as antigens in diagnostics for acquired immunodeficiency syndrome.19873312636
in vivo and in vitro host range of autographa californica nuclear polyhedrosis virus and spodoptera frugiperda nuclear polyhedrosis virus. 19873320215
analysis of the polyhedrin gene promoter of the autographa californica nuclear polyhedrosis virus.the polyhedrin gene promoter of the autographa californica nuclear polyhedrosis virus was analysed with respect to which sequences are required upstream of the mrna transcription initiation (cap) site for efficient promoter activity. insertions (8, 95 and 785 nucleotides) were made in this region at an ecor v site between the caat- and tata-like boxes. when these mutations were introduced into the virus they did not affect the activity of the polyhedrin promoter as judged by expression of the be ...19873320964
the development of multiple expression vectors for high level synthesis of eukaryotic proteins: expression of lcmv-n and acnpv polyhedrin protein by a recombinant baculovirus.a copy of the polyhedrin gene promoter of autographa californica nuclear polyhedrosis virus (acnpv), in association with the coding region of lymphocytic choriomeningitis virus n protein (lcmv-n) and the relevant polyhedrin transcription termination signals, has been cloned into the unique ecorv site of a plasmid representing an ecori derived fragment of the acnpv genome. the cloning site is upstream (but in the opposite orientation) of the natural acnpv polyhedrin gene. the derived pacvc2 trans ...19873334094
active human erythropoietin expressed in insect cells using a baculovirus vector: a role for n-linked oligosaccharide.biologically active recombinant human erythropoietin has been expressed at high levels in an insect cell background. expression involved the preparation of a human erythropoietin cdna, the transfer of this cdna to the autographa californica nuclear polyhedrosis virus (acnpv) genome under the polyhedrin gene promoter, and the subsequent infection of spodoptera frugiperda cells with recombinant acnpv. erythropoietin cdna was prepared through the expression of the human erythropoietin gene in cos c ...19872960381
activation of an insect baculovirus promoter in mammalian cells by adenovirus functions.the insect baculovirus autographa californica nuclear polyhedrosis virus (acnpv) replicates in insect cell lines in culture. in mammalian cells, however, the virus cannot be propagated. acnpv dna does not replicate or persist and is not transcribed in mammalian cells (tjia et al., 1983). in insect cells productively infected with acnpv, at least two major late viral gene products have been recognized, the polyhedrin, which makes up the bulk of the polyhedral inclusion bodies in infected cell nuc ...19872963453
effects of serial passage of autographa californica nuclear polyhedrosis virus in cell culture.a study of the major genomic alterations occurring during serial passage of autographa californica nuclear polyhedrosis virus (acnpv) in a trichoplusia ni cell line was conducted. progeny viruses from 24 independent passages were randomly selected and analyzed with restriction endonucleases. specific deletion mutations were generated repeatedly in the psti-g (7.6 to 13.1%) and the psti-i (14.4-17.9%) regions; these mutations became predominant in the serially passaged stocks in which they arose. ...19872887077
identification and nucleotide sequence of the regions of autographa californica nuclear polyhedrosis virus genome carrying insertion elements derived from spodoptera frugiperda.a morphology mutant of the baculovirus autographa californica nuclear polyhedrosis virus called m5 was previously shown to synthesize two size classes of viral dna, one of which had a deletion of 42% of the genome. it was hypothesized that the presence of the smaller m5 circular dna resulted from the specific deletion of the region located between two sites carrying short dna insertions. these sites have now been identified by dna-dna hybridization using cloned ecori fragments of m5 dna. one clo ...19872823472
location, transcription, and sequence of a baculovirus gene encoding a small arginine-rich polypeptide.the nucleotide sequence of the cdna of an abundant late 0.5-kilobase transcript of autographa californica nuclear polyhedrosis virus revealed a small open reading frame encoding an arginine-rich 6.9-kilodalton protein. the predicted amino acid composition of the 6.9-kilodalton protein was essentially identical to that of the core protein of viral nucleocapsids. the precise location of the 5' and 3' ends of the transcript were confirmed by s1 nuclease and primer extension analyses. multiple overl ...19873543402
baculovirus interaction with nontarget organisms: a virus-borne reporter gene is not expressed in two mammalian cell lines.the safety of baculoviruses with respect to mammalian species was studied by using a genetically engineered recombinant of autographa californica nuclear polyhedrosis virus. this recombinant contains the chloramphenicol acetyltransferase (cat) gene under the control of a mammalian-active promoter and expresses substantial levels of cat activity on infection of permissive and nonpermissive insect cells (l.f. carbonell, m.j. klowden, and l.k. miller, j. virol. 56:153-160, 1985). extremely low leve ...19873116925
abundant synthesis of functional human t-cell leukemia virus type i p40x protein in eucaryotic cells by using a baculovirus expression vector.the human t-cell leukemia virus type i (htlv-i) p40x protein is a 40-kilodalton polypeptide encoded in the 3'-terminal region of the virus. this protein is responsible for positive transcriptional trans-activation of promoter elements located within the htlv-i long terminal repeat. we introduced the protein-coding region of htlv-i p40x into the genome of the baculovirus autographa californica nuclear polyhedrosis virus. after infection of the insect spodoptera frugiperda (sf9) cell line, this re ...19873027397
production and characterization of the neutralization antigen vp2 of bluetongue virus serotype 10 using a baculovirus expression vector.dna representing rna segment 2 of bluetongue virus (btv) serotype 10, corresponding to the gene that codes for the btv neutralization antigen vp2, has been inserted into a baculovirus transfer vector in lieu of the 5' coding region of the polyhedrin gene of autographa californica nuclear polyhedrosis virus (acnpv). after cotransfection of spodoptera frugiperda cells with wild-type acnpv dna in the presence of the derived recombinant transfer vector dna, polyhedrin-negative recombinant baculoviru ...19873029984
synthesis and immunogenicity of the rotavirus major capsid antigen using a baculovirus expression system.rotaviruses are the major pathogens that cause life-threatening diarrhea in young children and animals. we inserted a simian rotavirus sa11 gene 6 cdna into the genome of the baculovirus autographa californica nuclear polyhedrosis virus adjacent to the strong polyhedrin promoter. the major capsid antigen (vp6) was expressed in high yields (20 to 150 micrograms/10(6) cells) when spodoptera frugiperda cells were infected with baculovirus recombinants containing sa11 gene 6 inserts. reactivity with ...19873033276
expression of polyomavirus large t antigen by using a baculovirus vector.a gene encoding the large t antigen of polyomavirus was inserted into the baculovirus autographa californica nuclear polyhedrosis virus so that gene expression was under the control of the strong, very late polyhedrin gene promoter. significantly more large t antigen was produced in recombinant virus-infected insect cells than was observed in polyomavirus-transformed mouse cells. the insect-derived t antigen exhibited polyomavirus origin-specific dna binding. the baculovirus expression system pr ...19873033300
a baculovirus vector can express intron-containing genes.a simjan virus 40 genomic fragment containing the genes coding for the large t and small t antigens was inserted into the genome of the baculovirus autographa californica nuclear polyhedrosis virus downstream of the strong polyhedrin promoter. infection of eucaryotic spodoptera frugiperda (sf9) cells with this recombinant virus produced significant amounts of small t antigen and little or no large t protein. analysis by northern blotting and s1 nuclease digestion revealed correct and preferentia ...19873033307
expression of bluetongue virus group-specific antigen vp3 in insect cells by a baculovirus vector: its use for the detection of bluetongue virus antibodies.dna representing rna segment 3 of bluetongue virus (btv) serotype 17, corresponding to the gene that codes for a group-specific antigen vp3, has been inserted into a baculovirus transfer vector in lieu of the 5' coding region of the polyhedrin gene of autographa californica nuclear polyhedrosis virus (acnpv). after cotransfection of spodoptera frugiperda cells with wild-type acnpv dna in the presence of the derived recombinant transfer vector dna, polyhedrin-negative recombinant baculoviruses we ...19873035063
divergent transcription of early 35- and 94-kilodalton protein genes encoded by the hindiii k genome fragment of the baculovirus autographa californica nuclear polyhedrosis virus.the organization of viral genes within the 3.7-kilobase-pair hindiii-k/ecori-s region of the autographa californica nuclear polyhedrosis virus genome (85 to 88 map units) was determined by using a combination of nucleotide sequencing, transcriptional mapping, and in vitro translation of hybrid selected rna. two nonoverlapping genes, extending in opposite directions and encoding polypeptides with molecular weights of 35,000 and 94,000 (35k and 94k polypeptides), were identified. unspliced, messen ...19873035225
overlapping sets of viral rnas reflect the array of polypeptides in the ecori j and n fragments (map positions 81.2 to 85.0) of the autographa californica nuclear polyhedrosis virus genome.in several parts of the autographa californica nuclear polyhedrosis virus (acnpv) genome, nested sets of overlapping rnas with common 3' or 5' termini have been recognized. in the present report, the pattern of viral transcription and the arrangement of viral gene products in the region of 81.2 to 85.0 map units were investigated. in this segment of the acnpv genome, at least nine size classes of viral rna were identified which ranged in size from 1.3 kilobases (kb) to 4.6 kb and exhibited commo ...19873041026
nucleotide sequence and temporal expression of a baculovirus regulatory gene.the nucleotide sequence of a trans-activating regulatory gene (ie-1) of the baculovirus autographa californica nuclear polyhedrosis virus has been determined. this gene encodes a protein of 581 amino acids with a predicted molecular weight of 66,856. a dna fragment containing the entire coding sequence of ie-1 was inserted downstream of an rna promoter. subsequent cell-free transcription and translation directed the synthesis of a single peptide with an apparent molecular weight of 70,000. quant ...198716789264
effects of temperature and ph on survival of free nuclear polyhedrosis virus of autographa californica.the effects of temperature and low ph on replication and survival of nonoccluded autographa californica nuclear polyhedrosis virus were investigated. no virus replication or formation of polynuclear inclusion bodies occurred at 37 degrees c. the virus was immediately inactivated upon exposure to ph 2.0 and was inactivated within 1 h at ph 4.0. the virus titer slowly declined, a 3-orders of magnitude reduction in virus titer, at ph 5.0 during a 4-h exposure. virus survival at ph 6.0 was equal to ...198716347494
evidence for microfilament involvement in budded autographa californica nuclear polyhedrosis virus production.autographa californica nuclear polyhedrosis virus (acnpv) is a large, double-stranded dna virus that infects lepidopteran insects. in the course of infection two different forms of the virus are produced; one that serves to spread the infection from insect to insect and another that spreads the infection within the insect from cell to cell. the latter form matures by budding from the plasma membrane of infected cells. we have found that cytochalasins b and d prevent the production of infectious ...198718644552
a point mutation in the polyhedrin gene of a baculovirus, autographa californica mnpv, prevents crystallization of occlusion bodies.the polyhedrin gene region of the autographa californica nuclear polyhedrosis virus(acmnpv) morphology mutant m29 has been characterized by genetic and physical techniques. recombination analysis of mutants m29 and acm5polyl demonstrated that wild-type polyhedrin recombinants could be obtained and that the dna restriction patterns of the recombinant viruses were identical to wild-type acmnpv dna. marker rescue experiments using the wild-type acmnpv ecori i fragment indicated that the morphology ...198719731454
multiple early transcripts and splicing of the autographa californica nuclear polyhedrosis virus ie-1 gene.the immediate-early ie-1 gene of autographa californica nuclear polyhedrosis virus was cloned, and its nucleotide sequence was determined. sequence analysis indicated that this gene would encode a protein of 582 amino acids with a predicted molecular weight of 66,822. analysis of ie-1 gene expression during baculovirus infection identified two transcripts. one, 1.9 kilobases (kb), was expressed at constant steady-state levels throughout infection, whereas the other, 2.1 kb, was expressed only ea ...19883043024
resistance of the 64k protein of budded autographa californica nuclear polyhedrosis virus to functional inactivation by proteolysis.the 64k surface protein of budded autographa californica nuclear polyhedrosis virus (acmnpv bv) is known to play a role in the functional entry of acmnpv bv into spodoptera frugiperda iplb-sf-21 cells by adsorptive endocytosis. acv1, a neutralizing monoclonal antibody, reacts with the 64k protein and in doing so prevents efficient entry. in this communication we report that treatment of acmnpv bv with either trypsin or proteinase k cleaves the 64k protein into one major fragment of 34.6k and two ...19883046120
co-expression of the hepatitis b surface and core antigens using baculovirus multiple expression vectors.the hepatitis b (hb) virus dna sequences coding for the pre-core (prec) or c antigens (hbpcag, hbcag) have been inserted into the baculovirus plasmid transfer vector, pacym1, such that the hb viral sequences are under the control of the polyhedrin promoter of autographa californica nuclear polyhedrosis virus (acnpv). spodoptera frugiperda cells infected with either of the derived recombinant plasmids in the presence of infectious acnpv dna yielded recombinant, polyhedrin-negative viruses that ex ...19883053987
association of autographa californica nuclear polyhedrosis virus (acmnpv) with the nuclear matrix.nuclear matrices from uninfected spodoptera frugiperda cells and those infected with autographa californica nuclear polyhedrosis virus (acmnpv) were isolated and their protein constituents were compared. proteins were characterized according to size and several different antibodies to drosophila nuclear proteins were employed in an attempt to identify the proteins comprising this nuclear substructure. three species of lamins were identified as major constituents of the nuclear matrix of spodopte ...19883055664
alteration of a lepidopteran peritrophic membrane by baculoviruses and enhancement of viral infectivity.the peritrophic membrane (pm), which lines the midgut of many insect species, has several functions. in particular, it may serve as a mechanical barrier to invading microorganisms. the protein composition of the pm from healthy and baculovirus-treated trichoplusia ni (cabbage looper) larvae was analyzed by polyacrylamide gel electrophoresis. a specific interaction took place between baculoviruses and the pm of susceptible t. ni larvae. a 68-kda glycoprotein of the pm disappeared within 15 min po ...19883055665
p39, a major baculovirus structural protein: immunocytochemical characterization and genetic location.a series of monoclonal antibodies was produced against proteins from polyhedra-derived virions of the multicapsid nuclear polyhedrosis virus of orgyia pseudotsugata (opmnpv). two of these antibodies (214 and 236) reacted with a protein of 39 kda on western blots of electrophoretically separated opmnpv viron proteins derived from both budded and polyhedra-derived virions. this protein appears to be a major component of both bv and pdv. one of the p39 antibodies was used to characterize p39 synthe ...19883059676
the location, sequence, transcription, and regulation of a baculovirus dna polymerase gene.the autographa californica nuclear polyhedrosis virus (acmnpv) dna polymerase gene was identified with the aid of an oligonucleotide probe corresponding to an amino acid sequence conserved among viral dna polymerases of other virus families. a 3.6-kb pair region of the acmnpv dna, from 39.5 to 42.5 map units (m.u.), was sequenced and an open reading frame (orf) of 2994 bp was observed. from the first atg of this orf, a translation product of 984 amino acids (mr 114,310) was predicted. amino acid ...19883059678
eight base pairs encompassing the transcriptional start point are the major determinant for baculovirus polyhedrin gene expression.the effects of mutations within the 92-bp region immediately upstream from the translational initiation atg of the polyhedrin gene of the baculovirus autographa californica nuclear polyhedrosis virus (acmnpv) were determined by plasmid transient expression assays in the presence of wild-type (wt) acmnpv dna. clustered point mutations were generated by substitution of 10-bp stretches of the polyhedrin promoter/leader region with a 10-bp hindiii linker. three of these linker scan (ls) mutations in ...19883071492
secretion of glycosylated human recombinant haptoglobin in baculovirus-infected insect cells.human haptoglobin (hp alpha 2 beta) was synthesized in insect cells using the baculovirus autographa californica nuclear polyhedrosis virus (acnpv) as an expression vector. viruses carrying the prohp alpha 2 beta cdna, either fused or non fused to viral polyhedrin dna sequences, expressed intracellularly low levels of unglycosylated and non maturated haptoglobin. on the contrary, recombinant viruses containing the preprohp alpha 2 beta cdna directed the expression of high levels of prohaptoglobi ...19883078448
functional studies on the p10 gene of autographa californica nuclear polyhedrosis virus using a recombinant expressing a p10-beta-galactosidase fusion gene.the beta-galactosidase gene (lacz) of escherichia coli was inserted in phase with the coding sequence of the autographa californica nuclear polyhedrosis virus (acmnpv) late-expressed mr 10,000 (p10) gene. the fusion gene was inserted into the acmnpv genome by cotransfection of a recombinant plasmid pacr159z, consisting of the ecori p fragment-containing pbr325-derived plasmid pacr159 and the lacz insert in the p10 gene, and wild-type acmnpvdna. infection of spodoptera frugiperda cells by the res ...19883128641
signals important for high-level expression of foreign genes in autographa californica nuclear polyhedrosis virus expression vectors.the transcriptional and translational signals required for efficient expression of the chloramphenicol acetyltransferase, beta-galactosidase, and tissue plasminogen activator genes, under the control of the polyhedrin promoter in spodoptera frugiperda cells infected with autographa californica nuclear polyhedrosis virus, were investigated by sds-page and rna dot blot analysis. the recombinant baculoviruses all contained alterations in the leader sequence or 5' proximal coding region of the polyh ...19883142147
human tissue-type plasminogen activator synthesized by using a baculovirus vector in insect cells compared with human plasminogen activator produced in mouse cells.a cdna fragment encoding the human tissue-type plasminogen activator was inserted into the baculovirus autographa californica nuclear polyhedrosis virus downstream from the polyhedrin promoter. the induction kinetics of t-pa was followed, after infection of spodoptera frugiperda cells, at both mrna and protein levels. fibrinolytically active plasminogen activator accumulated in the culture medium and reached 2.5 micrograms/ml after 120 h. the protein was compared with recombinant plasminogen act ...19883149610
[the autographa californica nuclear polyhedrosis virus as a eukaryotic vector system]. 19883150619
synthesis of a gene coding for an insect-specific scorpion neurotoxin and attempts to express it using baculovirus vectors.we have explored the possibility of improving baculovirus pesticides by incorporating an insect-specific neurotoxin gene into a baculovirus genome. a 112-bp gene (beit) encoding insectotoxin-1 of the scorpion buthus eupeus was synthesized and cloned in escherichia coli. for expression, beit was transferred to the dna genome of autographa californica nuclear polyhedrosis virus (acmnpv). three different recombinant acmnpvs, carrying beit under the control of the strong acmnpv polyhedrin promoter, ...19883243439
functional mapping of an acnpv immediately early gene which augments expression of the ie-1 trans-activated 39k gene.an early gene which augments the expression of the delayed early/late 39k gene of the baculovirus autographa californica nuclear polyhedrosis virus (acnpv) was identified by functional mapping. transient expression of the plasmid p39cat, containing the bacterial chloramphenicol acetyltransferase coding sequences under the control of the promoter of the 39k protein, was observed in cells cotransfected with acnpv dna digested with several restriction endonucleases. however, when p39cat was cotrans ...19883277328
a phosphorylated 34-kda protein and a subpopulation of polyhedrin are thiol linked to the carbohydrate layer surrounding a baculovirus occlusion body.surrounding baculovirus occlusion bodies is an electron-dense layer reported to be composed of carbohydrate which we term calyx. incubation of autographa californica nuclear polyhedrosis virus occlusion bodies (acmnpv obs) with dilute alkaline saline (das) followed by centrifugation at 12,000 g resulted in the sedimentation of calyx material which contained pp34, residual polyhedrin (p32), and entrapped occluded virions (das p-12 fraction). incubation of the das p-12 fraction with sodium dodecyl ...19883279702
autographa californica mnpv nucleocapsid assembly: inhibition by cytochalasin d.autographa californica nuclear polyhedrosis virus (acmnpv) is a large, double-stranded dna virus of lepidopteran insects. it replicates in the nucleus of host cells and produces two different infectious forms. one of them buds from the plasma membrane and is responsible for systemic infection in insects and for the spread of infection in cell culture. earlier we reported that cytochalasin d (cd) inhibited the production of this form of the virus in infected iplb-sf-21 cells by a mechanism other ...19883281373
glycosylation and processing of high levels of active human glucocerebrosidase in invertebrate cells using a baculovirus expression vector.a human cdna containing the complete coding region for the lysosomal glycoprotein glucocerebrosidase (ec 3.2.1.45) was introduced into the genome of autographa californica nuclear polyhedrosis virus downstream from the polyhedrin promoter. infection of spodoptera frugiperda cells (sf9) with recombinant virus produced high levels of glucocerebrosidase, 40% of which was in the culture medium. the amino-terminal amino acid sequence of the recombinantly produced enzyme was identical to that of matur ...19883282855
rate of increase and critical amount of nuclear polyhedrosis virus in lepidopterous larvae estimated from survival time assay data with a birth-death model.a birth-death model developed for pathogens of vertebrates was used to estimate the in vivo rate of increase (alpha) and the doubling time (td) from survival time assay data. host-pathogen combinations used in this study were two autographa californica nuclear polyhedrosis virus isolates in trichoplusia ni and heliothis zea npv in h. zea. the alpha's, estimated as he negative reciprocal of the slope of the linearly decreasing section of the plot of median survival times against the logarithm of ...19883291823
characterization of an early gene accelerating expression of late genes of the baculovirus autographa californica nuclear polyhedrosis virus.the region of the autographa californica nuclear polyhedrosis virus (acmnpv) encompassing the ecori t fragment (29.0 to 30.1 map units) was characterized by dna sequencing, transcriptional mapping, and site-directed mutagenesis. the largest transcript from this region, an early 1.7-kilobase (kb) poly(a)+ rna, encompassed three tandem, nonoverlapping open reading frames (orfs). the largest of these orfs, etl, was proximal to the 5' end of the transcript and had the capacity to encode a 28-kilodal ...19883292791
comparisons of host cell dna insertions and altered transcription at the site of insertions in few polyhedra bacilovirus mutants.few polyhedgra (fp) mutants of autographa californica nuclear polyhedrosis virus (acnpv) and the closely related strain galleria mellonella (gm)npv have been reported which contain trichoplusia ni host cell dna sequences inserted into the viral genome between map units 35.0 and 37.7. new fp mutants are described with alterations of the hindiii-i restriction enzyme fragment (33.8 to 37.7 map units) of acnpv, either deletions of viral dna sequences or insertions of spodoptera frugiperda host cell ...19882829419
functional analysis of the p10 gene 5' leader sequence of the autographa californica nuclear polyhedrosis virus.deletions were made in the 5' non-coding (leader) sequence of the autographa californica nuclear polyhedrosis virus (acnpv) p10 gene which progressively removed nucleotides upstream from the atg translation initiation codon. the effect of these deletions on p10 gene expression was studied using a transient expression assay. fragments containing the putative promoter and the entire or partly deleted 5' leader sequence of the p10 gene were inserted in front of the chloramphenicol acetyltransferase ...19882836797
expression of hepatitis a virus capsid sequences in insect cells.a cdna coding for hepatitis a virus (hav) vp1 and portions of the flanking vp3 and p2 sequences was inserted into the genome of autographa californica nuclear polyhedrosis virus under the control of the polyhedrin promoter and translational start codon. cells infected with recombinant virus produced high levels of a 55 kda protein, identified as containing hav vp1 by reactivity with anti-vp1 serum. the expressed protein also reacted on immunoblots with human hav convalescent sera as well as sera ...19882842974
synthesis of functional human immunodeficiency virus tat protein in baculovirus as determined by a cell-cell fusion assay.the human immunodeficiency virus tat protein is a strong trans-activator of the expression of mrnas originating from the viral long terminal repeat. we have expressed the first 72 amino acids (coding exon 1) of this protein in eucaryotic spodoptera frugiperda sf9 cells by using a baculovirus vector, autographa californica nuclear polyhedrosis virus. we show that the baculovirus vector stably produced the 72-amino-acid form of the tat protein but was unable to stably synthesize a larger 101-amino ...19882843682
bluetongue virus tubules made in insect cells by recombinant baculoviruses: expression of the ns1 gene of bluetongue virus serotype 10.bluetongue virus (btv) forms tubules in mammalian cells. these tubules appear to be composed of only one type of protein, ns1, a major nonstructural protein of the virus. to obtain direct evidence for the origin of the tubules, the complete m6 gene of btv serotype 10 was inserted into the baculovirus transfer vector pacym1, so that it was under the control of the polyhedrin promoter of autographa californica nuclear polyhedrosis virus. after cotransfection of spodoptera frugiperda cells with wil ...19882845115
overproduction of the protein encoded by the maize transposable element ac in insect cells by a baculovirus vector.the polypeptide encoded in the activator (ac) element of zea mays l. has been expressed in spodoptera frugiperda insect cells using plasmids which carry the strong polyhedrin promoter of the baculovirus autographa californica nuclear polyhedrosis virus (acnpv). recombinant acnpvs with the ac-cdna integrated and under the control of the viral polyhedrin promoter have been isolated and their genomes have been partly characterized as to the location of the foreign dna insert. upon infection of s. f ...19882851092
baculovirus expression of the small genome segment of hantaan virus and potential use of the expressed nucleocapsid protein as a diagnostic antigen.autographa californica nuclear polyhedrosis virus (acnpv) was used as an expression vector for the nucleocapsid protein gene of hantaan virus. two different cdna clones representing the small genome segment of hantaan virus were inserted into the transfer vector pacym1, and recombinants were generated by replacement of a portion of the baculovirus polyhedrin gene with the foreign, hantaan virus gene. recombinants containing both the first and second atg initiation codons of the hantaan virus gen ...19882895799
expression of human tyrosine hydroxylase cdna in invertebrate cells using a baculovirus vector.a human cdna containing the complete coding sequence for a human tyrosine hydroxylase (ec 1.14.16.2, form 2) was introduced into the genome of autographa californica nuclear polyhedrosis virus (acnpv) downstream to the polyhedrin promoter. infection of spodoptera frugiperda cells (sf9) with recombinant virus resulted in the expression of human tyrosine hydroxylase in these invertebrate cells. characterization of tyrosine hydroxylase activity in infected sf9 cells demonstrated both substrate and ...19882896667
purification and functional properties of simian virus 40 large and small t antigens overproduced in insect cells.the insect baculovirus autographa californica nuclear polyhedrosis virus was used as an expression vector for the simian virus 40 (sv40) small t (t) and large t (t) antigens. spodoptera frugiperda (sf9) cells infected with recombinant viruses encoding these proteins produced approximately 1 to 2 micrograms of t and up to 30 micrograms of t per 3 x 10(6) cells. the former was highly soluble after nonidet p-40 extraction of the infected cells, unlike its escherichia coli-produced counterpart. both ...19882969056
synthesis, purification, and characterization of the cytoplasmic domain of the human insulin receptor using a baculovirus expression system.the cytoplasmic domain of the beta subunit of the human insulin receptor has been overexpressed in insect cells using the baculovirus expression system. a recombinant baculovirus (bir-2) was constructed by inserting the human insulin proreceptor cdna fragment that encodes the cytoplasmic domain of the receptor into the genome of autographa californica nuclear polyhedrosis virus adjacent to the strong polyhedrin promoter. synthesis of the protein (baculovirus insulin receptor kinase (birk), mr 48 ...19882451671
regulation of host rna levels during baculovirus infection.during infection of the permissive host insect cell line spodoptera frugiperda iplb-sf-21 by the baculovirus autographa californica nuclear polyhedrosis virus (acmnpv), the levels of host actin, histone, and heat shock 70 (hsp70) rnas are reduced substantially. reduction of the host rna levels occurs primarily during a narrow window of the replication process, from approximately 12 to 18 hr postinfection (p.i.), corresponding to the phase in which the extracellular form of the virus buds into th ...19882459844
high level expression of nonfused foreign genes with autographa californica nuclear polyhedrosis virus expression vectors.high levels of nonfused chloramphenicol acetyltransferase, beta-galactosidase, and beta-glucuronidase expressed under the control of new vector constructs of the polyhedrin promoter in spodoptera frugiperda cells infected with autographa californica nuclear polyhedrosis virus were investigated by sds-page and rna dot blot analysis of total cytoplasmic rna. when the polyhedrin atg start codon was converted to att by site-directed mutagenesis, translation initiated at downstream atg codons resulti ...19892497580
a baculovirus blocks insect molting by producing ecdysteroid udp-glucosyl transferase.the predicted amino acid sequence of a newly identified gene of the insect baculovirus autographa californica nuclear polyhedrosis virus was similar to several uridine 5'-diphosphate (udp)-glucuronosyl transferases and at least one udp-glucosyl transferase. genetic and biochemical studies confirmed that this gene encodes an ecdysteroid udp-glucosyl transferase (egt). this enzyme catalyzes the transfer of glucose from udp-glucose to ecdysteroids, which are insect molting hormones. expression of t ...19892505387
a baculovirus gene with a novel transcription pattern encodes a polypeptide with a zinc finger and a leucine zipper.an autographa californica nuclear polyhedrosis virus gene encoding a 30-kilodalton polypeptide with two different sequence motifs characteristic of dna-binding proteins was identified immediately downstream of the major capsid protein gene (vp39). the gene, cg30, was characterized by sequencing, transcriptional mapping, in vitro translation of hybrid-selected rna, and comparison of the derived polypeptide sequence with published data bases. the initial atg of the 792-base-pair cg30 open reading ...19892507791
synthesis of immunogenic, but non-infectious, poliovirus particles in insect cells by a baculovirus expression vector.a baculovirus expression vector (acleon) derived from autographa californica nuclear polyhedrosis virus (acnpv) was prepared containing the complete 6.6 kb coding region of the p3/leon/37 strain of poliovirus type 3 placed under the control of the acnpv polyhedrin promoter. the recombinant virus was used to infect spodoptera frugiperda insect cells. as demonstrated by use of the appropriate antibodies, infected insect cells made poliovirus proteins that included the structural proteins vp0, vp1 ...19892543785
expression of largest rna segment and synthesis of vp1 protein of bluetongue virus in insect cells by recombinant baculovirus: association of vp1 protein with rna polymerase activity.the bluetongue virus core particles have been shown to contain an rna-directed rna polymerase (1). to identify the protein responsible for the virion rna polymerase activity, the complete 3.9 kb dna clone representing the largest rna segment 1 (l1) of bluetongue virus (btv-10) was placed under control of the polyhedrin promoter of autographa californica nuclear polyhedrosis virus (acnpv). the derived recombinant virus was used to infect spodoptera frugiperda cells. as demonstrated by stained pol ...19892552409
synthesis of functional bovine leukemia virus (blv) p34tax protein by recombinant baculoviruses.the bovine leukemia virus (blv) p34tax (also called tat, p34, xlor gene product) is a 34-kda polypeptide encoded in the 3'-terminal region of the virus. this protein is responsible for positive transcriptional trans-activation of promoter elements located within the blv long-terminal repeat. we introduced the protein-coding region of blv p34tax into the genome of the baculovirus autographa californica nuclear polyhedrosis virus. after infection of the insect spodoptera frugiperda (sf9) cell line ...19892554576
expression of hemolytically active human complement component c1r proenzyme in insect cells using a baculovirus vector.the gene of human c1r has been expressed in a baculovirus-insect-cell system via the pac373 transplacement vector. the full-length cdna copy was inserted into the pac373 vector downstream from the strong polyhedrin promoter of the baculovirus, autographa californica nuclear polyhedrosis virus (acnpv). spodoptera frugiperda cells were cotransfected with the resultant plasmid, pacc1r, and the wild-type acnpv dna. recombinant viruses, which drove the expression of c1r protein, were selected by plaq ...19892557186
molecular analysis of the transcriptional regulatory region of an early baculovirus gene.transcription of the gene encoding a 35,000-molecular-weight protein (35k protein) from the ecori-s region (86.8 to 87.8 map units) of autographa california nuclear polyhedrosis virus (acmnpv) occurs early in infection and declines later. the region promoting the gene for the 35k protein, extending from 426 base pairs (bp) upstream to 12 bp downstream from the rna start site, was linked to the bacterial chloramphenicol acetyltransferase gene (cat) for analysis. cat expression was monitored in ce ...19892642976
expression of sindbis virus 26s cdna in spodoptera frugiperda (sf9) cells, using a baculovirus expression vector.to study protein processing in an insect spodoptera frugiperda (fall armyworm; sf9) cell line, a 26s cdna encoding the sequence of sindbis virus structural proteins (capsid protein, of 30 kilodaltons [kda]; p62 [the precursor of e3 and e2], of 62 kda; a 6-kda peptide; and the e1 protein, of 56 kda) was inserted into the genome of autographa californica nuclear polyhedrosis virus (acnpv) adjacent to the polyhedrin promoter. by immunoblot analysis with antisera directed against whole sindbis virus ...19892644447
identification and sequence analysis of a gene encoding gp67, an abundant envelope glycoprotein of the baculovirus autographa californica nuclear polyhedrosis virus.monoclonal antibodies with specificity for the abundant envelope surface glycoprotein (gp67) of autographa californica nuclear polyhedrosis virus (acmnpv) were used to screen a lambda gt11 expression library of acmnpv dna fragments. the gp67 gene was mapped to the left end of the ecori h fragment in a right-to-left orientation on the consensus map of acmnpv. a 2.1-kilobase transcript which hybridized to the region was first detected in cell extracts at 2 h postinfection; it peaked in abundance a ...19892644449
location and nucleotide sequence of the 25k protein missing from baculovirus few polyhedra (fp) mutants.wild-type and few polyhedra (fp) mutants of the baculovirus autographa californica nuclear polyhedrosis virus (acmnpv) were studied to identify and sequence the gene encoding the 25-kda (25k) protein normally present in acmnpv-infected spodoptera frugiperda cells but which is often missing from fp mutant-infected cells. our previous study had mapped two overlapping late transcripts to the insertion site of host cell dna within the hindiii-i fragment (33.8 to 37.7 map units) of wild-type acmnpv. ...19892644735
a baculovirus polyhedral envelope-associated protein: genetic location, nucleotide sequence, and immunocytochemical characterization.using a polyclonal mouse antiserum produced against purified virions of the multicapsid nuclear polyhedrosis virus of orgyia pseudotsugata (opmnpv), two immunoreactive lambda gtii clones were identified which contained nonoverlapping insert dnas which mapped to a single open reading frame (orf) in the hindiii-m fragment. analysis of nucleotide sequence data indicates that this orf encodes a protein with a mw of 32.4 kda. a trpe-p32 gene fusion containing the entire p32 orf was constructed, and t ...19892646825
a physical map of spodoptera littoralis b-type nuclear polyhedrosis virus genome.a physical map for the genome of spodoptera littoralis (egyptian cottonworm) nuclear polyhedrosis virus of b-type (s1mnpv-b) [cherry cl, summers md (1985) j invertebr pathol 46: 289-295] was prepared using the restriction endonucleases smai, sfii, noti, apai, kpni, bsteii, psti, saci, and hindiii. the size of the genome was estimated to 136 kbp for the morocco isolate (s1mnpv-m2) and to 135 kbp for the lyon isolate (s1mnpv-l4). hindiii-k was used as the zero-point of the map because it hybridize ...19892647060
expression and characterization of hepatitis b virus surface antigen polypeptides in insect cells with a baculovirus expression system.the baculovirus autographa californica nuclear polyhedrosis virus was used as an expression vector to produce hepatitis b virus surface antigen with and without the pre-s domain. the s gene product was expressed as both fusion and nonfusion polypeptides. no difference was observed in the posttranslational modification of the fusion and nonfusion polypeptides. the s proteins were not secreted into the medium but were inserted into the endoplasmic reticulum, glycosylated, and partially extruded in ...19892648022
synthesis of biologically active influenza virus hemagglutinin in insect larvae.the hemagglutinin of influenza (fowl plague) virus was expressed in larvae of heliothis virescens by using recombinant autographa californica nuclear polyhedrosis virus (acnpv) as a vector. animals were infected with the recombinant virus either by parenteral injection or by feeding. for oral uptake, recombinant virus occluded in polyhedra obtained from cultured spodoptera frugiperda cells after coinfection with authentic acnpv was used. immunohistological analyses of infected animals revealed t ...19892648023
the pathway of infection of autographa californica nuclear polyhedrosis virus in an insect host.an immunohistochemical study was conducted to detect the temporal infection sequence of autographa californica m nuclear polyhedrosis virus in trichoplusia ni larvae. staining patterns indicated that the initial infection occurred in the midgut, simultaneously in columnar epithelial and regenerative cells, but that subsequently this tissue recovered. a major envelope glycoprotein stained in a polar fashion when it was expressed in columnar epithelial cells, but not when expressed in other cells ...19892648574
identification, sequence, and transcriptional mapping of the major capsid protein gene of the baculovirus autographa californica nuclear polyhedrosis virus.the gene encoding the major capsid protein of the baculovirus autographa californica nuclear polyhedrosis virus (acmnpv) was identified, sequenced, and transcriptionally mapped. the location of the gene was determined by immunological screening of an expression library of acmnpv open reading frame-beta-galactosidase fusions with an antibody raised to virus structural proteins. the dna sequence of the corresponding region, which mapped within 56.6 and 58.0 map units on the acmnpv genome, revealed ...19892649691
insertion of the sfmnpv polyhedrin gene into an acmnpv polyhedrin deletion mutant during viral infection.the autographa californica nuclear polyhedrosis virus (acmnpv) polyhedrin deletion mutant, d10a, was cotransfected with the recombinant plasmid sfmnpv.hindiii-l, which contains the spodoptera frugiperda nuclear polyhedrosis virus (sfmnpv) polyhedrin gene. an occlusion-positive hybrid virus was obtained which contained a dna insertion at approximately 13 map units on the acmnpv.d10a genome. a fine structure restriction map of cloned viral dna fragments from this region revealed that most of the 1 ...19892655273
effect of cytochalasin d on cell morphology and acmnpv replication in a spodoptera frugiperda cell line.cytochalasin d (cd) is a specific inhibitor of actin microfilament elongation and has been used to identify actin-dependent cellular processes. in this study we observed the effects of this inhibitor on autographa californica m nuclear polyhedrosis virus infected and uninfected iplb-sf-21 cells by electron microscopy. the cytochalasin d-induced morphological effects detected in uninfected cells included lobulate nuclei, double nuclei, long retraction processes, increased zeiosis, more frequent p ...19892656869
location, sequence, transcriptional mapping, and temporal expression of the gp64 envelope glycoprotein gene of the orgyia pseudotsugata multicapsid nuclear polyhedrosis virus.the gene encoding gp64, the envelope glycoprotein of the budded virus (bv) of orgyia pseudotsugata multicapsid nuclear polyhedrosis virus (opmnpv), was mapped to the hindiii-e fragment of of the viral genome and expression of the gp64 gene was examined at various times postinfection. to locate the gp64 gene, a cross-reacting monoclonal antibody (acv5) (a. w. hohmann and p. faulkner, 1983, virology, 125, 432-444) directed against the gp64 protein of the autographa californica multicapsid nuclear ...19892658304
conservation of polyhedrin gene promoter function between autographa californica and mamestra brassicae nuclear polyhedrosis viruses.the dna sequence of the polyhedrin gene of the mamestra brassicae multiple nucleocapsid nuclear polyhedrosis virus (mbmnpv) was determined and compared with the polyhedrin genes of autographa californica (ac) and panolis flammea (pf) mnpvs. using this information, a transfer vector was constructed based on the ecori i fragment of acmnpv in which the polyhedrin promoter was replaced by the homologous region extending 481 nucleotides upstream from the mbmnpv polyhedrin coding sequence. the escheri ...19892658411
expression of human plasminogen cdna in a baculovirus vector-infected insect cell system.a cdna that encodes the human plasminogen (hpg) amino acid sequence has been inserted adjacent to the polyhedrin promoter in the genome of the baculovirus, autographa californica nuclear polyhedrosis virus, which was then used to infect cultured cells of the farm armyworm, spodoptera frugiperda. under the conditions of cell growth employed, recombinant (rec)-hpg was secreted into the medium after 24 h postinfection (p.i.), at which point virtually no rec-hpg antigen remained inside the cells. at ...19892658810
a cytopathological investigation of autographa californica nuclear polyhedrosis virus p10 gene function using insertion/deletion mutants.the role of the autographa californica nuclear polyhedrosis virus p10 gene in viral cytopathology and morphogenesis was examined using classes of p10 deletion mutants with and without lacz (beta-galactosidase) gene fusion. mutant-infected cells did not form the fibrillar cytoplasmic and nuclear structures normally observed late in infection with wild-type (wt) virus, and the cells failed to lyse even at 2 weeks post-infection. based on wt and mutant cytopathology, we suggest lysis may be facilit ...19892659726
analysis of the promoter of the autographa californica nuclear polyhedrosis virus p10 gene.functional analyses of the p10 gene promoter from the autographa californica nuclear polyhedrosis virus (acnpv) were performed by progressively deleting the 230 nucleotides upstream from the p10 coding sequences towards the atg codon. truncated promoter sequences retaining the full 5' non-coding leader of p10 were inserted in front of the chloramphenicol acetyltransferase (cat) gene, and promoter activity in transfected acnpv-infected cells was measured using the transient cat expression assay. ...19892659727
analysis of transcription initiation in the panolis flammea nuclear polyhedrosis virus polyhedrin gene.the nucleotide sequence of the polyhedrin gene of panolis flammea multiple nucleocapsid polyhedrosis virus (pfmnpv) has been determined. the coding sequences of this gene shared 82% similarity at the dna level and 88% similarity at the protein level with the polyhedrin gene from autographa californica (ac) mnpv. a single nucleotide deviation from the consensus transcription initiation sequence for baculovirus very late genes was identified in the pfmnpv polyhedrin gene. rna was prepared from mam ...19892659729
engineering of an oryctes baculovirus recombinant: insertion of the polyhedrin gene from the autographa californica nuclear polyhedrosis virus.a cotransfection method has been developed for the generation of recombinant oryctes baculoviruses. the permissive coleopteran cell-line dsir-ha-1179 was transfected with a mixture of oryctes baculovirus dna (strain pv505) and a transfer vector. the transfer vector, a puc8-based plasmid, contained the polyhedrin gene from the autographa californica nuclear polyhedrosis virus, flanked by oryctes baculovirus dna from the hindiii fragment n. oryctes baculovirus does not form plaques with dsir-ha-11 ...19892659730
studies on the control region of the p10 gene of the autographa californica nuclear polyhedrosis virus.5' deletion mutants of the autographa californica nuclear polyhedrosis virus very late p10 gene promoter have been prepared and subjected to a transient expression assay in infected spodoptera frugiperda cells. the control plasmid contained the chloramphenicol acetyltransferase (cat) reporter gene under the control of the p10 promoter, which was included in a 230 bp sequence upstream from the p10 translation initiation codon. the control plasmid also contained a segment of the hr5 enhancer downs ...19892659731
characterization of the genetic organization of the hindiii m region of the multicapsid nuclear polyhedrosis virus of orgyia pseudotsugata reveals major differences among baculoviruses.a region including the 4 kb hindiii m fragment of the multicapsid nuclear polyhedrosis virus (mnpv) of orgyia pseudotsugata (opmnpv) genome was sequenced, transcriptionally mapped, and compared to the homologous region in the mnpv of autographa californica (acmnpv). five open reading frames (orfs) oriented in the same direction were identified and were found to be expressed at late times post-infection. the mrnas from the five orfs were found to coterminate at a single site downstream of orf 5. ...19892661722
fatty acid acylation of the 67k envelope glycoprotein of a baculovirus: autographa californica nuclear polyhedrosis virus.modification of autographa californica multicapsid nuclear polyhedrosis virus extracellular virion polypeptides with fatty acid was investigated. fatty acid-derived radioactivity was incorporated into gp67, the predominant virion envelope glycoprotein, during metabolic labeling using [3h]palmitic acid or [3h]myristic acid. the acyl moiety resisted extraction with chloroform-methanol but was released by mild alkali or hydroxylamine treatment, features characteristic of an ester-type linkage. palm ...19892672565
baculovirus polyhedrin promoter-directed expression of rubella virus envelope glycoproteins, e1 and e2, in spodoptera frugiperda cells.to study the capability of spodoptera frugiperda (fall armyworm; sf9) cells to synthesize and process mature rubella virus (rv) proteins, a cdna encoding the structural envelope glycoproteins, e1 (58 kda) and e2 (42-47 kda) were inserted into the genome of autographa californica nuclear polyhedrosis virus (acnpv) and expressed during infection under the transcriptional regulation of the polyhedrin gene promoter. by immunoblot analysis with antibodies directed against purified rv, the individual ...19892672567
sequence, transcription and translation of a late gene of the autographa californica nuclear polyhedrosis virus encoding a 34.8k polypeptide.a 1.4 kb region downstream of the dna polymerase gene of autographa californica nuclear polyhedrosis virus was sequenced. two open reading frames (orfs) were identified of 927 and 474 bases in length. the 927 base orf encodes a 34.8k protein as determined by in vitro translation of both hybrid-selected rna and rna synthesized in vitro from a 927 base orf template. the predicted amino acid sequence of the 34.8k polypeptide (p34.8) reveals a hydrophobic n terminus, two potential n-glycosylation si ...19892674327
expression and characterization of the ha-ras p21 protein produced at high levels in the insect/baculovirus system.normal and mutated cdnas of ha-ras have each been cloned into a standard (pac373) and a novel (p36c) baculovirus transfer vector and introduced via homologous recombination into the genome of autographa californica nuclear polyhedrosis virus immediately downstream of the polyhedrin promoter. spodoptera frugiperda cells infected with recombinant virus containing the normal ha-ras gene express very high levels of ras p21 protein (approximately 20% of total cell protein), whereas the mutant protein ...19892681210
cell-surface expression and purification of human cd4 produced in baculovirus-infected insect cells.cd4 is an integral membrane glycoprotein that acts as the cellular receptor for human immunodeficiency virus (hiv). a cdna encoding full-length cd4 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 cd4 as evaluated by flow cytometry and immunoblot analysis. recombinant cd4 expressed on the surfa ...19892682621
construction and analysis of an autographa californica nuclear polyhedrosis virus mutant lacking the polyhedral envelope.an autographa californica nuclear polyhedrosis virus (acmnpv)-specific protein of 34 kda (pp34) is shown to be involved in the morphogenesis of the polyhderal envelope of baculoviruses. the region of the acmnpv genome encompassing ecori-h and -s (map positions 82.6-85.8) contains five open reading frames (orfs) forming one transcriptional unit. the bacterial beta-galactosidase (lacz) gene was inserted in phase with the n-terminal 12 amino acids of orf3, thereby intervening this gene. a recombina ...19892683364
identification of p74, a gene essential for virulence of baculovirus occlusion bodies.dna sequencing of the hindiii-p fragment of the baculovirus autographa californica nuclear polyhedrosis virus downstream of a major late protein, p10, revealed the presence of an open reading frame (orf) 1935 nucleotides in length and in opposite polarity to p10. the gene product is considered essential for virus virulence in trichoplusia ni larvae since infection with occlusion bodies from a mutant, ac228z, in which portions of adjacent carboxy-termini from peptides p74 and p10 were deleted, fa ...19892688302
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