Publications
| Title | Abstract | Year(sorted ascending) Filter | PMID Filter |
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| expression in baculovirus vector system of the nucleocapsid protein gene of rinderpest virus. | the rinderpest (rv) nucleocapsid (np) gene segment was inserted into the genome of autographa californica nuclear polyhedrosis virus (acnpv) adjacent to the polyhedrin promoter. the expression of np protein in sf9 cells was confirmed by indirect immunofluorescence and by western blotting analysis with monoclonal antibodies. recombinant rv-np protein was purified by ultracentrifugation on a sucrose density gradient, and used as an antigen for an enzyme linked immunosorbent assay to detect anti rv ... | 1993 | 8366167 |
| the hr5 transcriptional enhancer stimulates early expression from the autographa californica nuclear polyhedrosis virus genome but is not required for virus replication. | autographa californica nuclear polyhedrosis virus (acmnpv) contains five homologous regions (hr1 through hr5) interspersed throughout its genome. analysis of plasmid transfections indicates that the hrs function as transcriptional enhancers and possible origins of viral dna replication. the role of these repetitive elements in regulating expression from the acmnpv genome was examined by constructing a series of recombinant viruses that tested the effect of hr5 on expression of the adjacent p35 g ... | 1993 | 8371342 |
| alternative transcriptional initiation as a novel mechanism for regulating expression of a baculovirus trans activator. | in this report, we show that the orgyia pseudotsugata nuclear polyhedrosis virus p34 gene, which is homologous to the autographa californica nuclear polyhedrosis virus pe-38 gene, is a trans activator. the predicted p34 protein contains a number of motifs that are similar to those found in other eukaryotic transcriptional trans activators, including a putative zinc finger dna-binding domain, a glutamine-rich domain, and a leucine zipper. northern (rna) blot analysis showed that the p34 gene is e ... | 1993 | 8371344 |
| the 1,629-nucleotide open reading frame located downstream of the autographa californica nuclear polyhedrosis virus polyhedrin gene encodes a nucleocapsid-associated phosphoprotein. | a 78-kda protein was produced in bacteria from a clone of the 1,629-nucleotide open reading frame located immediately downstream from the polyhedrin gene of autographa californica nuclear polyhedrosis virus. the identity of this protein was confirmed by its reactivity with peptide antiserum and amino terminal peptide sequencing after purification from transformed bacteria. the polypeptide was used to produce polyclonal antisera in rabbits. immunoblot analysis of insect cells infected with the ba ... | 1993 | 8371345 |
| sequence and temporal appearance of the early transcribed baculovirus gene he65. | we have identified the early transcribed he65 gene by screening a cdna library from polyadenylated rna which was isolated at 1 h after infection of spodoptera frugiperda cells with autographa californica nuclear polyhedrosis virus (acnpv). nucleotide sequencing analysis of the he65-specific cdna clone reveals one open reading frame of 1,662 nucleotides from which a protein of 65 kda in size can be predicted. the he65 gene is located downstream of the late transcribed p80 gene and upstream of the ... | 1993 | 8371346 |
| characterization of the replication of plasmids containing hr sequences in baculovirus-infected spodoptera frugiperda cells. | the replication of a series of plasmids, each containing one or more of 6 homologous regions (hrs) from the autographa californica multinucleocapsid nuclear polyhedrosis virus (acmnpv) genome, was investigated in acmnpv-infected spodoptera frugiperda cells. although a construct containing hr2 showed the most efficient replication, the sequences flanking the hr region appeared to influence the levels of replication. plasmids with non-hr-containing acmnpv inserts showed greatly reduced replication ... | 1993 | 8372444 |
| adenine dna methyltransferase m.cviri expression accelerates apoptosis in baculovirus-infected insect cells. | the adenine dna methyltransferase m.cviri (tgcma) gene from chlorella virus xz-6e was cloned into the autographa californica nuclear polyhedrosis virus (acmnpv) genome and expressed in spodoptera frugiperda insect cells under the control of two tandemly arranged viral promoters, the early etl promoter and the late polyhedrin promoter. m.cviri activity was first detected at 10 hr p.i and reached a maximum at 48 hr p.i. viral dna synthesized in insect cells infected with m.cviri expressing virus ( ... | 1993 | 8372450 |
| a gene encoding a highly expressed spindle body protein of heliothis armigera entomopoxvirus. | the gene encoding the most abundant protein of purified preparations of heliothis armigera entomopoxvirus (haepv) has been cloned and sequenced. the gene sequence encodes a 40.1k polypeptide with a putative n-terminal 20 amino acid leader peptide, and a single potential n-glycosylation site. analysis of the protein, which has an apparent m(r) of 50k on polyacrylamide gels, confirmed post-translational loss of the leader peptide, but showed no evidence of glycosylation. the protein is related to ... | 1993 | 8376960 |
| expression of bovine vitamin k-dependent carboxylase activity in baculovirus-infected insect cells. | a vitamin k-dependent carboxylase has recently been purified from bovine liver microsomes and candidate cdna clones have been isolated. definitive identification of the carboxylase remains circumstantial since expression of candidate carboxylase cdnas in mammalian cells is confounded by the presence of endogenous carboxylase activity. to overcome this problem, a recombinant strain of baculovirus (autographa california nuclear polyhedrosis virus, acmnpv) encoding a putative carboxylase (vbcbx/acm ... | 1993 | 8378308 |
| a 37-kilodalton glycoprotein from a baculovirus of orgyia pseudotsugata is localized to cytoplasmic inclusion bodies. | the gene encoding a 37-kda glycoprotein (gp37) of orgyia pseudotsugata multinucleocapsid nuclear polyhedrosis virus (opmnpv) was located and sequenced. gp37 of opmnpv was found to have 62 and 37% amino acid sequence identity with gp37 of autographa californica multinucleocapsid nuclear polyhedrosis virus (acmnpv) and with a protein reported to be a component of occlusion bodies from choristoneura biennis entomopoxvirus, respectively. the mrna start site of the opmnpv gp37 gene was mapped within ... | 1993 | 8380088 |
| development of baculovirus triple and quadruple expression vectors: co-expression of three or four bluetongue virus proteins and the synthesis of bluetongue virus-like particles in insect cells. | baculovirus multiple gene transfer vectors pacab3 and pacab4 have been developed to facilitate the insertion of three or four foreign genes respectively into the autographa californica nuclear polyhedrosis virus (acnpv) genome by a single co-transfection experiment. the pacab3 vector contains a polyhedrin promoter and two p10 promoters on either side of the polyhedrin promoter but in opposite orientations. the pacab4 vector has an additional polyhedrin promoter in opposite orientation to the fir ... | 1993 | 8385313 |
| the structural and functional organization of the autographa californica nuclear polyhedrosis virus genome. | baculoviruses are used as biological control agents of insect pests in agriculture and forestry. the multiple-nucleocapsid nuclear polyhedrosis virus of autographa californica (acmnpv) is the prototype baculovirus. recently, this virus has become widely used as vector for the high-level expression of foreign genes in insect cells. an updated physical map of restriction sites as well as the location of open reading frames (orfs) and transcripts are presented. most characteristic is the dispersal ... | 1993 | 8389114 |
| nucleotide sequence of the ubiquitin-39k gene region from the orgyia pseudotsugata multinucleocapsid nuclear polyhedrosis virus genome. | the region of the orgyia pseudotsugata multinucleocapsid nuclear polyhedrosis virus (opmnpv) genome containing the ubiquitin and the nuclear matrix-associated protein (39k) genes was sequenced. the first 77 amino acids of the opmnpv ubiquitin open reading frame (orf) showed 84.4% and 79% amino acid sequence identity to autographa californica multinucleocapsid nuclear polyhedrosis virus (acmnpv) and the spodoptera frugiperda insect ubiquitin, respectively. the predicted opmnpv ubiquitin protein c ... | 1993 | 8389803 |
| host range expansion of autographa californica nuclear polyhedrosis virus (npv) following recombination of a 0.6-kilobase-pair dna fragment originating from bombyx mori npv. | we have isolated hybrid baculoviruses of bombyx mori nuclear polyhedrosis virus (bmnpv) and autographa californica npv (acnpv) capable of replicating in both bmn (not susceptible to acnpv) and sf-21 (not susceptible to bmnpv) cells (a. kondo and s. maeda, j. virol. 65:3625-3632, 1991). repeated backcross infection of one of these recombinant isolates with acnpv generated eh-acnpv, a virus with restriction endonuclease patterns of genomic dna nearly identical to those of acnpv but capable of repl ... | 1993 | 8396678 |
| immunocytochemical characterization of p24, a baculovirus capsid-associated protein. | an open reading frame (orf 1) located upstream of the polyhedron envelope protein gene in the orgyia pseudotsugata multinucleocapsid nuclear polyhedrosis virus (opmnpv) genome was cloned in-frame into a trpe bacterial expression vector. the fusion protein produced by this construct was used for the preparation of a monospecific antiserum. western blot analysis of extracts from opmnpv-infected lymantria dispar cells and autographa californica npv (acmnpv)-infected spodoptera frugiperda cells dete ... | 1993 | 8423444 |
| bovine respiratory syncytial virus nucleocapsid protein expressed in insect cells specifically interacts with the phosphoprotein and the m2 protein. | the coding region of the gene for the nucleocapsid (n) protein of bovine respiratory syncytial virus has been inserted into the genome of autographa californica nuclear polyhedrosis virus (acnpv) using transfer vector pvl 1393. infection of spodoptera frugiperda cells with recombinant virus resulted in the synthesis of high levels of n protein. this protein was indistinguishable from the authentic bovine respiratory syncytial virus (brsv) n protein by sds-gel electrophoresis and immunoprecipitat ... | 1993 | 8438581 |
| nucleotide sequence of the p74 gene of a baculovirus pathogenic to the spruce budworm, choristoneura fumiferana multicapsid nuclear polyhedrosis virus. | polypeptide p74 has been found to be essential for production of virulent occlusion bodies of the baculovirus autographa californica multicapsid nuclear polyhedrosis virus (acmnpv). hybridization with acmnpv-derived probes has led to the location of the p74 gene in the spruce budworm virus, choristoneura fumiferana mnpv. sequence data indicate that cfmnpv p74 is 73% identical to acmnpv at the nucleotide level and 77% identical at the amino acid level. elements of predicted secondary structure ar ... | 1993 | 8439559 |
| three baculovirus genes involved in late and very late gene expression: ie-1, ie-n, and lef-2. | we developed a method to identify baculovirus genes required for late and very late gene expression that is based on subtraction of clones from an autographa californica nuclear polyhedrosis virus genomic library which is able to trans activate promoters of reporter plasmids in transient expression assays. using this assay, we found that three genes located between 83.7 and 7.5 map units of the autographa californica nuclear polyhedrosis virus genome were involved in expression from the late cap ... | 1993 | 8445724 |
| an apoptosis-inhibiting baculovirus gene with a zinc finger-like motif. | spodoptera frugiperda sf-21 cells infected with autographa californica nuclear polyhedrosis virus mutants which lack a functional p35 gene undergo apoptosis, a type of programmed cell death. to identify p35-homologous genes in other baculoviruses, a. californica nuclear polyhedrosis virus dna containing a deletion in p35 was cotransfected into sf-21 cells along with genomic dnas from other baculoviruses. one of the viral dnas which were able to rescue wild-type infection was from cydia pomonella ... | 1993 | 8445726 |
| nucleotide sequence and transcriptional analysis of a gene encoding gp41, a structural glycoprotein of the baculovirus autographa californica nuclear polyhedrosis virus. | 1993 | 8445739 | |
| ligand binding by a recombinant insect juvenile hormone binding protein. | a cdna for the hemolymph juvenile hormone binding protein (jhbp) of larval manduca sexta has been isolated, sequenced, and expressed in an insect cell line. a recombinant baculovirus, containing the jhbp cdna fused to the p10 promoter of autographa californica nuclear polyhedrosis virus, was constructed. insect cells (sf9) infected with this virus secreted recombinant jhbp (rjhbp) into the medium (> 50 micrograms/ml), and cotranslational removal of an 18 amino acid leader sequence was observed. ... | 1993 | 8448166 |
| expression of hornet genes encoding venom allergen antigen 5 in insects. | antigen 5, also known as dol m v, is a major allergen found in the venom of the baldfaced hornet, dolichovespula maculata. we have inserted the f10 and f17 cdnas, which encode hornet antigen 5 (ha5) forms 2 (dol m; vb) and 3 (dol m va), respectively, into the genome of the baculovirus, acmnpv, to produce the recombinant baculovirus gene expression vectors, vev-ha5f10 and vev-ha5f17. insect cells infected with either vev-ha5f10 or vev-ha5f17 produce and secrete a novel protein with an electrophor ... | 1993 | 8467098 |
| functional domains of the p10 protein of autographa californica nuclear polyhedrosis virus. | distinct functional domains in the autographa californica nuclear polyhedrosis virus p10 protein were identified by analysis of p10 mutants. when up to 15 amino acids from the carboxy terminus were deleted, truncated p10 proteins were found in both the nucleus and the cytoplasm of infected cells, but formed no fibrillar structures. this suggested that the positively charged carboxy terminus is not required for nuclear or cytoplasmic localization of p10 protein, but is involved in protein-protein ... | 1993 | 8468550 |
| sequence, transcriptional mapping, and overexpression of p47, a baculovirus gene regulating late gene expression. | a 2.8-kb region of the autographa californica nuclear polyhedrosis virus genome was sequenced and found to contain an open reading frame (p47) which was capable of rescuing a previously characterized temperature-sensitive mutant, ts317 (s. partington, h. yu, a. lu, and e. b. carstens, virology 157:91-102, 1990). transcriptional mapping demonstrated that an early 4.2-kb rna encoded the p47 open reading frame and probably overlapped the 39k delayed-early gene. the p47 open reading frame was cloned ... | 1993 | 8474157 |
| the 35-kilodalton protein gene (p35) of autographa californica nuclear polyhedrosis virus and the neomycin resistance gene provide dominant selection of recombinant baculoviruses. | autographa californica nuclear polyhedrosis virus (acmnpv) recombinants were constructed to test the effectiveness of the acmnpv 35-kilodalton protein gene (35k gene) and the bacterial neomycin resistance gene (neo) as dominant selectable markers for baculoviruses. insertion of the acmnpv apoptosis suppressor gene (p35) into the genome of p35-deletion mutants inhibited premature host cell death and increased virus yields up to 1200-fold at low multiplicities in spodoptera frugiperda (sf21) cell ... | 1993 | 8493093 |
| identification and characterization of lef-1, a baculovirus gene involved in late and very late gene expression. | an autographa californica nuclear polyhedrosis virus (acmnpv) gene required in transient expression assays for late and very late viral gene expression was identified, sequenced, and transcriptionally mapped. this gene, designated late expression factor 1 (lef-1), was located between 7.4 and 8.7 map units of the acmnpv physical map. it was identified by cotransfecting spodoptera frugiperda cultured cells with a collection of overlapping cloned dna fragments covering the entire acmnpv genome and ... | 1993 | 8497062 |
| efficient secretion of attacin from insect fat-body cells requires proper processing of the prosequence. | the attacins constitute a group of immune proteins induced after bacterial infection in the moth hyalophora cecropia. they are synthesized as preproproteins and undergo post-translational modification during transport to the hemolymph. the processing and transport rates of attacin were studied in its natural host as a response to infection. monensin totally inhibited the processing from proattacin to attacin and the radiolabeled proattacin remained intracellular. this observation indicated that ... | 1993 | 8508799 |
| nucleotide sequence and transcriptional analysis of the p10 gene of spodoptera exigua nuclear polyhedrosis virus. | the p10 gene of spodoptera exigua multiple nuclear polyhedrosis virus (semnpv) was localized on the xbai fragment h (5.1 kb) of the physical map of the viral genome. the coding sequence of the semnpv p10 gene is 264 nucleotides (nt) long corresponding to a predicted protein of 88 amino acids with an mhf of 9607. the semnpv p10 protein showed only limited amino acid identity (39% and 26%, respectively) to those of orgyia pseudotsugata mnpv (opmnpv) and autographa californica mnpv (acmnpv) and thu ... | 1993 | 8509757 |
| apoptosis reduces both the in vitro replication and the in vivo infectivity of a baculovirus. | apoptotic programmed cell death occurs when the insect cell line sf-21, derived from spodoptera frugiperda, is infected with mutants of the baculovirus autographa californica nuclear polyhedrosis virus (acmnpv) which lack a functional p35 gene. however, infection of the trichoplusia ni tn-368 cell line with p35 mutants does not result in apoptosis (r. clem, m. fechheimer, and l. miller, science 254:1388-1390, 1991). we have examined the effect of apoptosis on acmnpv infections in cell lines and ... | 1993 | 8510202 |
| in vitro transcription from baculovirus late gene promoters: accurate mrna initiation by nuclear extracts prepared from infected spodoptera frugiperda cells. | extracts prepared from nuclei of autographa californica nuclear polyhedrosis virus-infected spodoptera frugiperda cells were shown to support in vitro transcription from baculovirus late gene promoters. in vitro transcription was optimized for the late promoter of the 39k gene. the mg2+ concentration was critical; concentrations higher than 1 to 2 mm did not support late transcription. additional conditions included template (40 micrograms/ml), extract (2.5 mg/ml), and incubation time (25 min). ... | 1993 | 8510206 |
| a baculovirus-expressed fusion protein containing the antibody-binding domain of protein a and insect luciferase. | a fusion construct encoding two antibody-binding sites of protein a from staphylococcus aureus and click beetle, pyrophorus plagiophthalamus, luciferase (lucgr) was designed and expressed using the baculovirus system. the construct was inserted under the transcriptional regulation of the polyhedrin gene promoter of the autographa californica nuclear polyhedrosis virus (acnpv) and expressed in the insect spodoperta frugiperda cell line during viral infection. the properties of the resultant chime ... | 1993 | 8512706 |
| a method for producing recombinant baculovirus expression vectors at high frequency. | a system has been developed that can generate recombinant baculovirus expression vectors at frequencies approaching 100%. this system provides a selection for recombinant viruses by using the essential gene downstream of the autographa californica nuclear polyhedrosis virus (acmnpv) polyhedrin expression locus. two acmnpv derivatives were constructed in which the expression locus and part of the downstream gene are flanked by restriction sites. the parental viruses are viable; however, restricti ... | 1993 | 8512707 |
| a simple and efficient procedure for the oral inoculation of trichoplusia ni larvae with polyhedrin-negative recombinant baculovirus. | current procedures for inoculating lepidopteran larvae with polyhedrin-negative recombinant baculovirus, i.e. intracoelomic injection or coinfection with wild type virus, are laborious and can compromise final yields of recombinant protein. herein is described a simple and efficient method for oral inoculation. up to 100% infection was obtained when individual early fifth instar trichoplusia ni larvae were fed a small piece of a formaldehyde-free insect diet to which 4.2 x 10(5) pfu of a polyhed ... | 1993 | 8514841 |
| location of two putative origins of dna replication of autographa californica nuclear polyhedrosis virus. | previously, we described a defective autographa californica nuclear polyhedrosis virus (acmnpv) which must contain cis-acting elements required for dna synthesis, such as the origin(s) of replication (ori). defective genomes of acmnpv generated after serial undiluted passage were analyzed further. three small separated regions were retained in dna of defective acmnpv and accumulated in extracellular defective interfering viruses as well as in intracellular dna after 40 passages. two of these reg ... | 1993 | 8517035 |
| 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 |
| analyses of alternative poly(a) signals for use in baculovirus expression vectors. | synthetic, duplicated, or heterologous sequences encompassing putative 3' mrna processing (polyadenylation, poly(a)) sites have been analyzed with regard to their involvement in processing mrna species synthesized from the polyhedrin promoter of autographa californica nuclear polyhedrosis virus (acnpv). the data indicate that a synthetic sequence based on the 3' processing signal of the rabbit beta-globin gene is used more efficiently than an sv40-based, early gene poly(a) signal, or the acnpv 1 ... | 1993 | 8100374 |
| comparison of oligosaccharide processing among various insect cell lines expressing a secreted glycoprotein. | the processing of the n-linked oligosaccharide modifying a secreted alkaline phosphatase glycoprotein (seap) expressed with a recombinant autographa californica nuclear polyhedrosis virus was evaluated in insect cell lines established from spodoptera frugiperda, trichoplusia ni, and mamestra brassicae. studies with endoglycosidase h (endo h), which removes high-mannose oligosaccharides, revealed that 79% of the intracellular seap produced in the m. brassicae-derived mb0503 cell line was endo h r ... | 1993 | 8167899 |
| protein engineering studies on c1r and c1s. | 1. c1r and c1s cdnas were placed downstream the strong polyhedrin promoter in the autographa californica nuclear polyhedrosis virus and the recombinant proteins were expressed in insect cells, in biologically active form. the yield of expression is high enough to get recombinant components for chemical and functional studies (5 micrograms/ml cell culture supernatant). 2. the biological activity and the post-translational modifications of the recombinant subcomponents were checked. the rc1r and r ... | 1993 | 8172556 |
| penetration of autographa californica nuclear polyhedrosis virus nucleocapsids into iplb sf 21 cells induces actin cable formation. | the budded form of autographa californica nuclear polyhedrosis virus functionally enters cells by adsorptive endocytosis. during the period of virus uptake into spodoptera frugiperda iplb sf21 cells (1 to 4 hr postinfection), filamentous f-actin cables, visualized by fluorescence microscopy, were formed within the cytoplasm of infected cells. cable formation appeared to be a direct effect of viral inoculum in that the numbers of observed cables increased with an increase in multiplicity of infec ... | 1993 | 8212560 |
| characterization of productive and non-productive acmnpv infection in selected insect cell lines. | we have investigated the process of productive and non-productive acmnpv infection in a variety of cultured insect cells including lines derived from spodoptera frugiperda (iplb-sf21), mamestra brassicae (ses-mabr-3), choristoneura fumiferana (iprl-cf-1), bombyx mori (bmn-4), lymantria dispar (iplb-ld652y), helicoverpa zea (hz1b3), and drosophila melanogaster (dm). in each cell line, we have examined viral utilization of an early, a late, and a very late promoter, the replication of viral dna an ... | 1993 | 8212569 |
| expression of the human follicle-stimulating hormone receptor in the baculovirus system. | human follitropin receptor (hfshr) cdna was inserted into the genome of autographa californica nuclear polyhedrosis virus under the control of the polyhedrin promoter. spodoptera frugiperda (sf9) insect cells expressed the hfshr at the membrane level, 2-3 days after infection. ligand binding analysis demonstrated saturable, high-affinity binding of 125i-labeled recombinant hfsh (kd = 0.17 nm) as well as binding specificity for both human and equine fsh. interestingly, a highly purified preparati ... | 1993 | 8216320 |
| expression of full-length and truncated forms of crystal protein genes from bacillus thuringiensis subsp. kurstaki in a baculovirus and pathogenicity of the recombinant viruses. | full-length and truncated forms of the crystal protein gene cryia(b) derived from bacillus thuringiensis subsp. kurstaki hd-1 and full-length cryia(c) gene of b. thuringiensis subsp. kurstaki hd-73 were introduced into the genome of the baculovirus autographa californica nuclear polyhedrosis virus, in place of the polyhedrin gene. all gene constructs were expressed at high levels in insect cells and insects upon infection with the recombinant viruses. the protein products were shown to be biolog ... | 1993 | 8228317 |
| divergence and evolution of homologous regions of bombyx mori nuclear polyhedrosis virus. | homologous regions (hrs) (hr1,hr2-left,hr2-right,hr3,hr4-left,hr 4-right, and hr5) similar to those found in the autographa californica nuclear polyhedrosis virus (acnpv) genome were found in the bombyx mori npv (bmnpv) genome. the bmnpv hrs contained two to eight repeats of a homologous nucleotide sequence which were on average about 75 bp long. all of these homologous sequence repeats contained a 26-bp-long palindrome motif with an ecori or ecori-like site at its core. the consensus sequence o ... | 1993 | 8230471 |
| recombinant leishmania surface glycoprotein gp63 is secreted in the baculovirus expression system as a latent metalloproteinase. | a gene encoding the leishmania surface metalloproteinase, gp63, was modified using the polymerase chain reaction to obtain effective secretion of recombinant gp63 (regp63) in the baculovirus insect cell expression system. the coding region for the n-terminal signal peptide (sp) of gp63 was modified to resemble the sp for the gp67 envelope protein from the budded virus form of autographa californica nuclear polyhedrosis virus. to prevent processing at the c-terminus with a glycosyl phosphatidylin ... | 1993 | 8244034 |
| expression of the baculovirus p35 gene inhibits mammalian neural cell death. | expression of the apoptosis suppressor gene p35, derived from the baculovirus autographa californica nuclear polyhedrosis virus, markedly inhibited the cell death of stably transfected mammalian neural cells whether the cell death was induced by glucose withdrawal, calcium ionophore, or serum withdrawal. the p35 protein, which is required to block virus-induced apoptosis of cultured insect cells, is only the second gene product shown to block mammalian neural cell death, with bcl-2 being the fir ... | 1993 | 8245984 |
| identification of genes encoding late expression factors located between 56.0 and 65.4 map units of the autographa californica nuclear polyhedrosis virus genome. | using a previously developed method that allows the identification of autographa californica nuclear polyhedrosis virus (acmnpv) genes which stimulate transient expression from a late and a very late viral promoter [passarelli and miller, j. virol., 67, 2149-2158 (1993)], we have identified three genes between 56.0 and 65.4 map units of the acmnpv genome involved in expression from a late and a very late promoter but not from an early viral promoter. one gene, p143, was previously shown to be es ... | 1993 | 8249293 |
| identification and characterization of a putative origin of dna replication in the genome of a baculovirus pathogenic for orgyia pseudotsugata. | a four-kilobase (kb) region (hindiii-n, map units 7.0-11.3) of the orgyia pseudotsugata multinucleocapsid nuclear polyhedrosis virus (opmnpv) genome was found to contain sequences that conferred upon plasmids the ability to undergo infection-dependent replication. the plasmid dna appeared to be replicated to form high molecular weight multimers. plasmids with deletions of up to 1.8 kb from either end of the hindiii-n region were replication competent. however, a discrete sequence, contained with ... | 1993 | 8249294 |
| construction of a cdna encoding the multifunctional animal fatty acid synthase and expression in spodoptera frugiperda cells using baculoviral vectors. | a cdna encoding the 2505-residue multifunctional rat fatty acid synthase has been constructed and expressed as a catalytically active protein in spodoptera frugiperda (sf9) cells using autographa californica nuclear polyhedrosis virus (baculovirus). the 7.5 kb cdna was engineered by the amplification and sequential splicing together of seven fragments contained in overlapping cdnas that collectively spanned the entire rat fatty acid synthase coding sequence. the full-length cdna was cloned into ... | 1993 | 8250834 |
| ttaa serves as the target site for tfp3 lepidopteran transposon insertions in both nuclear polyhedrosis virus and trichoplusia ni genomes. | we have analysed tfp3 transposable elements from five independently isolated fp mutants of the autographa californica nuclear polyhderosis virus (acmnpv). we also analysed genomic copies of tfp3 elements amplified from the dnas of the trichoplusia ni cell line (tn-368) and t. ni larvae using the polymerase chain reaction (pcr). the sequences of all the newly isolated tfp3 elements closely resemble the previously described tfp3/1 element. each of the transposons isolated from the virus mutants du ... | 1993 | 8269089 |
| genetically engineered multi-component virus-like particles as veterinary vaccines. | multiprotein structures can be constructed to mimic virus particles. these engineered particles lack genetic material and are not infectious but they can elicit protective immune responses in animals against challenges with infectious viruses. as a prototype, insect cells were co-infected with two recombinant baculoviruses. one recombinant baculovirus contained an insert of l2 and m5 genes, which encode for outer capsid proteins vp2 and vp5, respectively, from bluetongue virus (btv) downstream o ... | 1993 | 8270267 |
| identification of seven putative origins of autographa californica multiple nucleocapsid nuclear polyhedrosis virus dna replication. | seven putative origins of dna replication (oris) were identified and located on the genome of autographa californica multiple nucleocapsid nuclear polyhedrosis virus (acmnpv), when an improved infection-dependent replication assay was used. a threefold higher yield of amplified plasmid was achieved when an m.o.i. of 1 was used (instead of 25), and another twofold increase was obtained when the interval between transfection and infection was extended from 5 to 24 h. six of the putative oris were ... | 1993 | 8277271 |
| baculovirus-directed expression of human prostatic steroid 5 alpha-reductase 1 in an active form. | in the human prostate, the enzyme steroid 5 alpha-reductase (h5 alpha r) catalyses the conversion of testosterone into the more potent androgen, dihydrotestosterone. two distinct cdnas coding for h 5 alpha r in the human prostate have been previously characterized. enzyme h5 alpha r1 shows a maximum activity at basic ph whereas h5 alpha r2 has an acidic ph optimum activity. we report here the expression of the human steroid h5 alpha r1 in a eukaryotic expression system: the baculovirus-directed- ... | 1993 | 8664165 |
| extension of autographa californica nuclear polyhedrosis virus host range by interspecific replacement of a short dna sequence in the p143 helicase gene. | recombinant baculoviruses obtained by coinfection of insect cells with autographa californica and bombyx mori nuclear polyhedrosis viruses (acnpv and bmnpv, respectively) possess a wider in vitro host range than either parent virus. to localize the dna sequences responsible for this species specificity, we used a two-step method of production and selection of recombinant viruses with altered specificity. sf9 cells, which are permissive for acnpv, were first cotransfected with genomic acnpv dna a ... | 1994 | 8278405 |
| a mutant baculovirus with a temperature-sensitive ie-1 transregulatory protein. | we have mapped the mutation responsible for the temperature-sensitive (ts) phenotype of tsb821, a mutant of the baculovirus autographa californica nuclear polyhedrosis virus (h. h. lee and l. k. miller, j. virol. 31:240-252, 1979), to a single nucleotide which changes alanine 432 of the multifunctional regulatory protein ie-1 to a valine. mapping was done with a combination of marker rescue and transient expression assays, hybrid gene construction by overlap pcr gene splicing, and nucleotide seq ... | 1994 | 8289337 |
| in vivo and in vitro analyses of recombinant baculoviruses lacking a functional cg30 gene. | the cg30 gene of autographa californica nuclear polyhedrosis virus (acmnpv) encodes two sequence motifs, a zinc finger-like motif and a leucine zipper, found in other polypeptides known to be involved in gene regulation. to gain insight into the function of the cg30 product, cg30, we constructed and characterized recombinant viruses lacking a functional cg30 gene. we found that cg30 mutants had no striking phenotype in cell lines derived from spodoptera frugiperda or trichoplusia ni or in t. ni ... | 1994 | 8289348 |
| high-level expression and purification of secreted forms of herpes simplex virus type 1 glycoprotein gd synthesized by baculovirus-infected insect cells. | two forms of herpes simplex virus glycoprotein gd were recombined into autographa californica nuclear polyhedrosis virus (baculovirus) and expressed in infected spodoptera frugiperda (sf9) cells. each protein was truncated at residue 306 of mature gd. one form, gd-1(306t), contains the coding sequence of patton strain herpes simplex virus type 1 gd; the other, gd-1(qaat), contains three mutations which eliminate all signals for addition of n-linked oligosaccharides. prior to recombination, each ... | 1994 | 8289380 |
| functional mapping of regions of the autographa californica nuclear polyhedrosis viral genome required for dna replication. | previous results showed that plasmids containing one of the eight putative origins (ori's) of autographa californica nuclear polyhedrosis virus (acmnpv) are replicated after transfection into spodoptera frugiperda cells if essential trans-acting factors are supplied by acmnpv infection (kool et al., virology, 192, 94-101, 1993a; kool et al., j. gen. virol., in press, 1993b; leisy and rohrmann, virology, 196, 722-730, 1993). in this report a transient complementation assay is described in which f ... | 1994 | 8291249 |
| functional dissection of the autographa california nuclear polyhedrosis virus enhancer element hr5. | the hr5 enhancer element of autographa californica nuclear polyhedrosis virus stimulates transcription from baculovirus-delayed early promoters in the presence of the viral transactivator, ie1. gel retardation analyses of interactions between a fragment of hr5 and extracts prepared from cells transfected with an ie1-encoding plasmid revealed the presence of three dna-protein complexes. in order to better define the functional domains of the hr5 enhancer, we constructed a set of plasmids containi ... | 1994 | 8178424 |
| identification of lef-7: a baculovirus gene affecting late gene expression. | we have examined a 15-kb region of the autographa californica nuclear polyhedrosis virus (acmnpv) genome, from 66 to 78 map units, for the presence of genes which transactivate expression from late and very late viral promoters in transient expression assays. one gene in this region activated reporter gene expression approximately six- to eightfold when the reporter gene was under the control of the late promoter of the major capsid protein gene, vp39, or the very late promoter of the polyhedrin ... | 1994 | 8178427 |
| specificity of baculovirus p10 functions. | three functional domains in baculovirus p10 proteins have been postulated for aggregation, nuclear disintegration, and fibrillar structure formation (van oers et al., j. gen. virol. 74, 563-574, 1993). to study the specificity of these functions, a recombinant autographa californica nuclear polyhedrosis virus (accr1) was constructed in which the coding sequence of the p10 gene was replaced with the p10 sequence of the distantly related spodoptera exigua (se) mnpv. in accr1-infected cells the sem ... | 1994 | 8178439 |
| secretion of recombinant rat annexin 5 by insect cells in a baculovirus expression system. | rat annexin 5 was expressed in insect cells using a baculovirus vector, autographa californica nuclear polyhedrosis virus. the rat annexin 5 cdna was prepared by a polymerase chain reaction using mrna from rat pituitary glands and placed under the control of the polyhedrin promoter. the gene product was 36 k dalton and was recognized by anti-rat annexin 5 serum. the calcium dependent binding of the recombinant annexin 5 to membranes was confirmed. the recombinant protein appeared in the medium b ... | 1994 | 8185595 |
| flow cytometric analysis of bioluminescence emitted by recombinant baculovirus-infected insect cells. | five recombinant baculoviruses, each containing a different insect luciferase gene encoding a protein with characteristic light emission properties, namely, luc gr (546 nm), luc ff (556 nm), luc yg (560 nm), luc ye (578 nm), and luc or (593 nm) were constructed. all genes were inserted under the transcriptional control of the polyhedrin gene promoter of the autographa californica nuclear polyhedrosis virus (acnpv) and expressed in spodoptera frugiperda insect cells during viral infection. the bi ... | 1994 | 8187580 |
| the apoptotic suppressor p35 is required early during baculovirus replication and is targeted to the cytosol of infected cells. | the p35 gene of autographa californica nuclear polyhedrosis virus (acmnpv) is required to block virus-induced apoptosis. the trans-dominant activity of p35 suppresses premature cell death and facilitates acmnpv replication in a cell line- and host-specific manner. to characterize the p35 gene product (p35), a specific polyclonal antiserum was raised. as revealed by immunoblot analyses of wild-type acmnpv-infected cells, p35 appeared early (8 to 12 h) and accumulated through the late stages of in ... | 1994 | 8189486 |
| detection of a putative 30-kda ligand of the cluster-2 antigen. | the cluster-2 antigen, also called egp-2, is a 38-kda transmembrane glycoprotein with a distribution that is largely confined to human epithelial cells and their derived carcinomas. monoclonal antibodies (mabs) directed against egp-2 have been extensively studied as anti-tumor agents, yet the function of the antigen is not known. in the present study we used a biotinylated recombinant soluble derivative of the egp-2 (segpbio) as a probe to detect a possible egp-2 ligand, using various carcinoma ... | 1994 | 8194899 |
| mutagenesis of recombinant protein c inhibitor reactive site residues alters target proteinase specificity. | protein c inhibitor (pci) is a heparin-binding plasma serine proteinase inhibitor (serpin) which is thought to be a physiological regulator of activated protein c. we are using recombinant pci (rpci) to study structural determinants of target proteinase specificity. a cdna encoding full-length pci has been expressed as a fully active proteinase inhibitor using autographa californica nuclear polyhedrosis virus (baculovirus). rpci was expressed maximally 4 days after infection and could be express ... | 1994 | 8206990 |
| identification and transcriptional regulation of the baculovirus lef-6 gene. | we identified an autographa californica nuclear polyhedrosis virus (acmnpv) gene, the late expression factor 6 gene (lef-6), which is involved in expression from late and very late acmnpv gene promoters but not from an early acmnpv gene promoter in transient expression assays. this gene was located within the psti i fragment of the acmnpv genome (14.7 to 17.9 map units), immediately downstream of ac-iap, the acmnpv homolog of a baculovirus gene family involved in blocking apoptotic programmed ce ... | 1994 | 8207819 |
| a baculovirus gene involved in late gene expression predicts a large polypeptide with a conserved motif of rna polymerases. | we have identified and sequenced a novel baculovirus gene, late expression factor eight gene (lef-8), of autographa californica nuclear polyhedrosis virus that is necessary for efficient expression from late and very late virus gene promoters in a transient expression assay. the predicted gene product, lef-8, has a molecular mass of 102 kda and contains a conserved sequence motif, gxkx4hgq/nkg, found in dna-directed rna polymerases throughout the animal, plant, and microbial kingdoms. | 1994 | 8207843 |
| nucleotide sequence and genetic organization of a 7.3 kb region (map unit 47 to 52.5) of autographa californica nuclear polyhedrosis virus fragment ecori-c. | the nucleotide sequence and genetic organization of a 7297 bp region within the ecori-c fragment of autographa californica multiple nucleocapsid nuclear polyhedrosis virus (acmnpv) are presented. eight putative open reading frames were found and their respective amino acid sequences compared with a number of data libraries. orf 1227 corresponded with gp41 and its predicted protein sequence was found to be 55 amino acids longer at its c terminus than reported previously. moreover the main part of ... | 1994 | 8126447 |
| deletion and mutational analyses of bluetongue virus ns2 protein indicate that the amino but not the carboxy terminus of the protein is critical for rna-protein interactions. | genome segment 8 (s8) of bluetongue virus serotype 10 (btv-10) encodes the nonstructural protein ns2. this protein, which has single-stranded rna (ssrna) binding capacity, is found in btv-infected cells in the form of virus inclusion bodies (vibs). to identify the domain(s) important for rna binding and oligomerization of the protein, a number of deletions were made in regions of the gene that code for either the amino or carboxy terminus of the protein. the modified genes were cloned into and e ... | 1994 | 8139002 |
| an apoptosis-inhibiting gene from a nuclear polyhedrosis virus encoding a polypeptide with cys/his sequence motifs. | two different baculovirus genes are known to be able to block apoptosis triggered upon infection of spodoptera frugiperda cells with p35 mutants of the insect baculovirus autographa californica nuclear polyhedrosis virus (acmnpv):p35 (p35-encoding gene) of acmnpv (r. j. clem, m. fechheimer, and l. k. miller, science 254:1388-1390, 1991) and iap (inhibitor of apoptosis gene) of cydia pomonella granulosis virus (cpgv) (n. e. crook, r. j. clem, and l. k. miller, j. virol. 67:2168-2174, 1993). using ... | 1994 | 8139034 |
| temporal regulation of a complex and unconventional promoter by viral products. | the dna polymerase (dnapol) gene of autographa californica nuclear polyhedrosis virus presents a complex promoter organization. it lacks the usual tata box and start site, and its rna accumulation initially increases and then decreases dramatically during infection. we investigated dnapol temporal regulation. transiently expressed dnapol gene was transcribed at a low level from minor start sites. coexpression with ie0 and/or ie1 immediate-early genes dramatically enhanced dnapol transcription, s ... | 1994 | 8139038 |
| mutational analysis of a baculovirus major late promoter. | we have used a linker-scan mutation strategy to analyze pcap99, the proximal promoter of the autographa californica nuclear polyhedrosis virus (acmnpv) gene encoding the major capsid protein. a series of recombinant viruses expressing the cat reporter gene under the control of selected mutants of this promoter was constructed. only mutations that altered bases within a region extending from 8 bp upstream to 6 bp downstream from a taag sequence had a significant effect on expression from the late ... | 1994 | 8144021 |
| human monocyte chemoattractant protein-1 expressed in a baculovirus system. | human monocyte chemoattractant protein-1 (hmcp-1) was produced using a baculovirus system. the hmcp-1 cdna was inserted into the genomic dna of autographa californica nuclear polyhedrosis virus (acnpv) using a transfer vector, pjvp10z. spodoptera frugiperda insect cells, which were infected with this recombinant virus, secreted recombinant hmcp-1 (re-hmcp-1) at the level of 10-20 micrograms/ml of culture medium. this product was shown to chemoattract monocytes. three distinct bands of 11, 11.5 a ... | 1994 | 8144036 |
| dengue type-2 virus envelope protein made using recombinant baculovirus protects mice against virus challenge. | the gene coding for the envelope (e) glycoprotein of dengue-2 virus was cloned into baculovirus (autographa californica nuclear polyhedrosis virus). the recombinant virus contained the entire e protein gene, preceded by 38 nucleotides from the end of the prematrix glycoprotein gene and followed by the first 83 nucleotides of nonstructural protein 1. when expressed in spodoptera frugiperda (sf9) cells, approximately 1 mg of recombinant e antigen was made per 10(9) cells. this antigen reacted with ... | 1994 | 8147490 |
| identification and characterization of the ecdysteroid udp-glucosyltransferase gene of the lymantria dispar multinucleocapsid nuclear polyhedrosis virus. | we have located, cloned, sequenced and characterized the ecdysteroid udp-glycosyltransferase gene (egt) gene from the baculovirus lymantria dispar multinucleocapsid nuclear polyhedrosis virus. (ldmnpv), which is specific for the gypsy moth (l. dispar). the egt gene from the related baculovirus autographa californica multinucleocapsid nuclear polyhedrosis virus (acmnpv) disrupts the hormonal balance of the host larva by galactosylating ecdysone, which prevents moulting. the location of the ldmnpv ... | 1994 | 8151299 |
| expression of recombinant rabbit udp-glcnac: alpha 3-d-mannoside beta-1,2-n-acetylglucosaminyltransferase i catalytic domain in sf9 insect cells. | udp-glcnac: alpha 3-d-mannoside beta-1,2-n-acetylglucosaminyltransferase i (gnt i; ec 2.4.1.101) catalyses a key reaction in the conversion of oligomannose to complex and hybrid n-glycans. the cytoplasmic tail and transmembrane segment of rabbit gnt i cdna were replaced with an in-frame cleavable signal sequence and the hybrid construct was inserted into the genome of autographa californica nuclear polyhedrosis virus (acmnpv) under the control of the polyhedrin promoter. sf9 insect cells were in ... | 1994 | 7841795 |
| the structure and function of the first component of complement: genetic engineering approach (a review). | the availability of cdna and genomic clones for the subcomponents of c1, as well as the recognition of the modular organization of serine-proteases have opened up exciting new possibilities for approaching structural problems. in this review the latest achievements of combined protein engineering, functional and structural studies are summarized. the concept of this research is to construct deletion, point and hybrid mutants of the highly homologous c1r and c1s subcomponents, to reveal the funct ... | 1994 | 7866721 |
| human 17 beta-hydroxysteroid dehydrogenase: overproduction using a baculovirus expression system and characterization. | estrogenic 17 beta-hydroxysteroid dehydrogenase (17 beta-hsd) plays a pivotal role in the synthesis of estrogens. we overproduced human placental estrogenic 17 beta-hsd using a baculovirus expression system for the study of the enzyme mechanism. a cdna encoding the entire open reading frame of human 17 beta-hsd was inserted into the genome of autographa californica nuclear polyhedrosis virus and expressed in spodoptera frugiperda (sf9) insect cells. metabolic labeling and western blot analysis u ... | 1994 | 7918113 |
| expression of semliki forest virus nsp1-specific methyltransferase in insect cells and in escherichia coli. | we have expressed the semliki forest virus (sfv)-specific nonstructural protein nsp1 both in insect cells and in escherichia coli in the absence of other viral proteins. a substantial amount of nsp1 was synthesized in sf9 cells infected with the recombinant autographa californica nuclear polyhedrosis virus (acnpv) acnpv-nsp1. these cells had a high level of guanine-7-methyltransferase activity compared with that of wild-type acnpv-infected cells. the methyltransferase activity and nsp1 were most ... | 1994 | 7933125 |
| cloning and expression of a soluble sialidase from chinese hamster ovary cells: sequence alignment similarities to bacterial sialidases. | a cdna encoding a soluble sialidase from chinese hamster ovary (cho) cells has been cloned and expressed. completely degenerate oligonucleotide primers, which were based on the amino acid sequence of peptides obtained from the purified sialidase (warner et al., glycobiology, 3, 455-463, 1993), and the polymerase chain reaction, with single-stranded cdna template, were employed to generate a unique oligonucleotide probe. the unique probe of 93 bp was used for screening a lambda gt 10 cho cell cdn ... | 1994 | 7949662 |
| the pk-1 gene of autographa californica multinucleocapsid nuclear polyhedrosis virus encodes a protein kinase. | open reading frame (orf) 9 of the ecori i fragment of the baculovirus autographa californica multinucleocapsid nuclear polyhedrosis virus (acmnpv) genome encodes a protein, pk-1, which has strong similarity to serine-threonine protein kinases. the published sequence of the pk-1 gene contains an error that, when corrected, extends the orf by 228 nucleotides. transcription of pk-1 was first detectable by primer extension at 12 h post-infection, accumulating to high levels during the very late phas ... | 1994 | 7964609 |
| suppression of apoptosis in insect cells stably transfected with baculovirus p35: dominant interference by n-terminal sequences p35(1-76). | expression of p35 from the dna genome of autographa californica nuclear polyhedrosis virus (acmnpv) suppresses virus-induced apoptosis and promotes virus replication in spodoptera frugiperda (sf21) cells. to examine the molecular mechanism by which p35 prevents apoptosis in insects, sf21 cells were stably transfected with p35. neomycin-resistant cell lines that synthesized protein p35 were identified. stable transfection with p35 protected sf21 cells from apoptosis induced by actinomycin d conce ... | 1994 | 7966562 |
| identification and characterization of vlf-1, a baculovirus gene involved in very late gene expression. | we have identified a gene required for strong expression of the polyhedrin gene by characterizing a mutant, tsb837, of the baculovirus autographa californica nuclear polyhedrosis virus (acmnpv) which is temperature sensitive (ts) for occluded virus production at the nonpermissive temperature. marker rescue experiments utilizing an overlapping set of acmnpv genomic clones revealed that the gene responsible for the ts mutant phenotype mapped to a region between 46 and 48 map units. fragments (2.2 ... | 1994 | 7966564 |
| biosynthesis and processing of the autographa californica nuclear polyhedrosis virus gp64 protein. | gp64 is a major virion envelope glycoprotein of the baculovirus autographa californica multicapsid nuclear polyhedrosis virus (acmnpv). gp64 plays an important role in acmnpv infection, probably mediating penetration of one form of the virus into host cells through the endocytic pathway. gp64 also represents an excellent probe for studying the membrane glycoprotein processing capabilities of baculovirus-infected insect cells, which are used widely as a eucaryotic expression system. the goals of ... | 1994 | 7975226 |
| functional characterization of partially purified epstein-barr virus dna polymerase expressed in the baculovirus system. | the dna polymerase gene of epstein-barr virus (ebv) was cloned into baculovirus transfer vector (pbluebac). the recombinant baculovirus (acebp-15) was obtained by cotransfection of spodoptera frugiperda (sf9) cells with infectious dna from autographa californica multiple nuclear polyhedrin virus (acmnpv) and pbluebac plasmid carrying ebv polymerase gene. infection of sf9 cells with the recombinant virus produced substantial quantities of the ebv dna polymerase protein of the expected size (110 k ... | 1994 | 7975269 |
| functional dissection of the ie2 gene product of the baculovirus autographa californica nuclear polyhedrosis virus. | the psti-n genomic fragment of autographa californica nuclear polyhedrosis virus (acnpv) encodes an immediate-early protein, ie2, that functions as a promiscuous transactivator of other early viral promoters and heterologous promoters both in vivo and in vitro. ie2 contains several sequence motifs that are common to transcriptional activators. we have employed site-directed mutagenesis coupled with transient-expression assays to identify the amino acid sequences of ie2 that are essential for tra ... | 1994 | 8009830 |
| autographa californica nuclear polyhedrosis virus, pdv, and ecv viral envelopes and nucleocapsids: structural proteins, antigens, lipid and fatty acid profiles. | autographa californica nuclear polyhedrosis virus (acmnpv) infection results in the production of two types of infectious, enveloped viruses. as both of these viral forms play significantly different roles in the virus life cycle, the different functional characteristics in the roles of the virus may be explained, in part, by the protein and lipid composition and source of the viral envelopes. both viruses utilize different maturation and envelopment strategies: extracellular virus (ecv) obtains ... | 1994 | 8009843 |
| reiterated dna fragments in defective genomes of autographa californica nuclear polyhedrosis virus are competent for acmnpv-dependent dna replication. | we previously reported on the generation of approximately 50-kb size defective genomes (dgs) which appeared to retain less than 2.2% of the standard autographa californica nuclear polyhedrosis virus (acmnpv) dna between 85.0 and 87.2 mu while the rest of the virus dna had been largely deleted (lee and krell, j. virol., 66:4339-4347, 1992). to investigate these presumably repeated sequences further, we cloned and analyzed the most abundant hypermolar 1.80-kb xhol dna fragment as well as a minor b ... | 1994 | 8009853 |
| engineering the assembly pathway of the baculovirus-insect cell expression system. | the synthesis of complex biological structures such as antibodies using recombinant dna technology is a major biotechnological advance. active murine antibody (igg) oligomers, composed of two heavy (h) and two light (l) polypeptide chains, have been expressed and secreted by the baculovirus-insect cell expression system. unfortunately, expression of the functional antibodies is accompanied by the formation of abnormal protein complexes and aggregates in which the polypeptide chains are bound tog ... | 1994 | 8010671 |
| expression of aleutian mink disease parvovirus capsid proteins in a baculovirus expression system for potential diagnostic use. | a 2.3-kb cdna clone encoding aleutian mink disease parvovirus (adv) structural proteins vp1 and vp2 was inserted into the polyhedron gene of autographa californica nuclear polyhedrosis virus (acnpv) and expressed by the recombinant virus, acadv-1, in spodoptera frugiperda-9 cells. sodium dodecyl sulfate-polyacrylamide gel electrophoresis and western immunoblot analysis (wia) indicated that synthesis of both vp1 and vp2 was being directed by acadv-1. fluorescence microscopic examination of acadv- ... | 1994 | 8011776 |
| superior expression of juvenile hormone esterase and beta-galactosidase from the basic protein promoter of autographa californica nuclear polyhedrosis virus compared to the p10 protein and polyhedrin promoters. | the expression characteristics of the p10, polyhedrin and basic protein promoters of autographa californica nuclear polyhedrosis virus were compared using two reporter enzymes, juvenile hormone esterase (jhe) and beta-galactosidase. in these systems, jhe is exported from the cell and beta-galactosidase is localized to the cytosol. expression of jhe from the basic, p10 and polyhedrin promoters was first detected in the medium at 13, 19 and 27 h post-infection respectively. the basic protein promo ... | 1994 | 8021586 |
| identification of the gp67 gene of a baculovirus pathogenic to the spruce budworm, choristoneura fumiferana multinucleocapsid nuclear polyhedrosis virus. | the baculovirus gp67 gene encodes a ph-dependent membrane fusion protein and has been identified in both autographa californica multinucleocapsid nuclear polyhedrosis virus (acmnpv) and orygia pseudotsugata mnpv (opmnpv). we have identified a homologous gene in the spruce budworm virus, choristoneura fumiferana mnpv (cfmnpv). the cfmnpv gp67 gene is 79% identical to acmnpv gp67 at the level of nucleotide sequence and 82% identical at the level of predicted amino acid sequence. as with opmnpv and ... | 1994 | 8021612 |
| genome organization of the dna-binding protein gene region of cryptophlebia leucotreta granulosis virus is closely related to that of nuclear polyhedrosis viruses. | the nucleotide sequence and genomic organization of a conserved genome region within the ecori g fragment of cryptophlebia leucotreta granulosis virus (clgv) is presented. five open reading frames (orfs) were identified which were homologous to those of autographa californica nuclear polyhedrosis virus (acmnpv), located upstream of the helicase gene (p143) at 63.6 to 65.6 map units. the orfs of clgv and acmnpv share nucleotide sequence homologies of about 47 to 53% and are very similarly arrange ... | 1994 | 8021613 |
| the complete dna sequence of autographa californica nuclear polyhedrosis virus. | the complete nucleotide sequence of the genome of clone 6 of the baculovirus autographa californica nuclear polyhedrosis virus (acnpv) has been determined. the molecule comprises 133,894 base pairs and has an overall a + t content of 59%. our analysis suggests that the virus encodes some 154 methionine-initiated, and potentially expressed, open reading frames (orfs) of 150 nucleotides or greater. these orfs are distributed evenly throughout the virus genome on either strand. the orfs are arrange ... | 1994 | 8030224 |
| the autographa californica nuclear polyhedrosis virus ie2 gene encodes a transcriptional regulator. | the baculovirus autographa californica nuclear polyhedrosis virus (acnpv) expresses several immediate early genes in infected cells. one of these, ie2 (formerly called ien), was mapped and identified by its ability to augment ie1-mediated transactivation of the delayed early 39k gene. ie2 also stimulated the expression of reporter genes linked to the ie1 promoter. to test whether the increase in expression was due to increased mrna, primer extension analyses were performed. spodoptera frugiperda ... | 1994 | 8030237 |
| expression and post-translational processing of a broad-spectrum organophosphorus-neurotoxin-degrading enzyme in insect tissue culture. | a recombinant baculovirus, autographa californica nuclear polyhedrosis virus (acnpv), has been utilized to express the opd (organophosphate-degrading) gene from pseudomonas diminuta in insect tissue-culture cells (sf9) of the fall armyworm (spodoptera frugiperda). the broad-spectrum organophosphate hydrolase (ec 3.1.8.1) encoded by this gene is a member of a general class of enzymes [organophosphate (op) anhydrolases] that include parathion hydrolases, di-isopropyl-fluorophosphatases (dfpases), ... | 1994 | 8031504 |
| control of programmed cell death by the baculovirus genes p35 and iap. | the sf-21 insect cell line undergoes rapid and widespread apoptosis when treated with actinomycin d or when infected with a mutant of the baculovirus autographa californica nuclear polyhedrosis virus lacking a p35 gene or a functionally active iap (inhibitor of apoptosis) gene. here we provide evidence that the basis for the induction of apoptosis by these two different stimuli is the cessation of rna synthesis. we also show that expression of either p35 or two different functional iap homologs ... | 1994 | 8035800 |
| characterization of glycosylated and catalytically active recombinant human alpha-galactosidase a using a baculovirus vector. | fabry disease is an x-linked inborn error of glycolipid metabolism caused by a deficiency of the lysosomal enzyme alpha-galactosidase a (gala; ec 3.2.1.22). in order to obtain large quantities of this human enzyme for physical characterization and for the development of new approaches for enzyme therapy, we constructed derivatives of the autographa californica nuclear polyhedrosis virus that produce the human enzyme. the recombinant gala (re-gala) is produced at high levels, and is active with b ... | 1994 | 8039705 |
| cloning and expression of bovine and porcine interleukin-2 in baculovirus and analysis of species cross-reactivity. | the cdnas encoding bovine and porcine interleukin-2 (il-2) have been expressed using the baculovirus autographa californica nuclear polyhedrosis virus as a vector in insect cells. insect cells infected with recombinant viruses secreted bovine and porcine il-2 into the culture medium, with biological activities for maintaining the proliferation of homologous cells. when the activities of these two il-2 proteins and commercially available human il-2 were tested on heterologous cells differences we ... | 1994 | 8042283 |
| interaction of trichoplusia ni granulosis virus-encoded enhancin with the midgut epithelium and peritrophic membrane of four lepidopteran insects. | enhancin, an infectivity-enhancing protein from trichoplusia ni granulosis virus (tngv) was tested for its ability to increase autographa californica multiple nucleocapsid nuclear polyhedrosis virus (acmnpv) infection in the larvae of four lepidopteran insects. enhancin increased the mortality of acmnpv infection in all the four insect species tested. peritrophic membrane (pm) assays showed altered protein profiles in pms treated with enhancin in all the four species. this supports the hypothesi ... | 1994 | 8046398 |