Publications
| Title | Abstract | Year(sorted ascending) Filter | PMID Filter |
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| nucleotide sequence analysis of the hcnpv cysteine protease gene. | the cysteine protease (cp) gene of hyphantria cunea nuclear polyhedrosis virus (hcnpv) has been located at the hind iii--3.5 kb fragment. the nucleotide sequence has been determined. the results indicated that its open reading frame of 975 nt encoded the protein of 324 amino acids. the baculovirus late gene promoter taag motif was identified at -24 nt upstream from the start codon atg; the typical poly (a) signal sequence aataaa was detected at 33 nt downstream from the stop codon taa. the hcnpv ... | 1998 | 12174267 |
| expression and cellular distribution of baculovirus-expressed bovine herpesvirus 1 (bhv-1) glycoprotein d (gd) sequences. | glycoprotein d (gd) of bovine herpesvirus 1 (bhv-1), a homolog of herpes simplex virus gd, represents a major component of the viral envelope and is a dominant immunogen. to study the antigenic properties of the different regions of gd, we have expressed the full-length gd encoding gene and overlapping fragments spanning various regions of the gd open reading frame in a baculovirus (autographa californica nuclear polyhedrosis virus)--insect cell (spodoptera frugiperda, sf-9) system. maximum leve ... | 1998 | 9856100 |
| molecular cloning and analysis of transcription initiation in the anagrapha falcifera multiple nucleocapsid polyhedrosis virus polyhedrin gene. | using autographa californica multiple nucleocapsid polyhedrosis virus (acmnpv) polyhedrin to probe the southern blots of anagrapha falcifera multiple nucleocapsid polyedrosis virus (afmnpv), we identified the location of the afmnpv polyhedrin gene within the 7.2 kb ecori fragment. the 7.2 kb ecori fragment of afmnpv was cloned and the nucleotide sequences of the polyhedrin coding region and its flanking regions were determined. nucleotide sequence analysis indicated the presence of an open readi ... | 1998 | 9856340 |
| recombinant bombyx mori nucleopolyhedrovirus harboring green fluorescent protein. | the gene encoding the green fluorescent protein (gfp) under the control of the highly expressed autographa californica nucleopolyhedrovirus (acmnpv)-polyhedrin promoter has been introduced into the polyhedrin (polh) locus of bombyx mori nucleopolyhedrovirus (bmnpv) by homologous recombination. the insect host larvae and the cultured cells infected with this recombinant virus (vbmgfp) showed high levels of expression of gfp. the larval tissues permissive to virus multiplication could be readily v ... | 1998 | 9863053 |
| molecular characterization, enzymatic analysis, and purification of murine proprotein convertase-1/3 (pc1/pc3) secreted from recombinant baculovirus-infected insect cells. | a cdna coding for the murine proprotein convertase-1 (mpc1 also known as mpc3 or mspc3) was inserted into the autographa californica nuclear polyhedrosis virus. following infection of spodoptera frugiperda cells, the recombinant n-glycosylated protein is secreted into the cell culture medium from which it can be purified to homogeneity as a fully enzymatically active enzyme. two major secreted molecular forms of mpc1 with apparent molecular weights of 85 and 71 kda, respectively, and a minor one ... | 1998 | 9882569 |
| an efficient way to introduce unique restriction endonuclease sites into a baculovirus genome. | recombinant baculoviruses which can be linearized at unique sites with restriction endonucleases can greatly facilitate the construction of other recombinants including baculovirus expression vectors and site-specific mutants. we designed a strategy to introduce unique restriction endonuclease sites at virtually any location in a baculovirus genome. the unique sites were first introduced onto a transfer plasmid which also contained in the vector portion of the plasmid an e. coli lacz gene and a ... | 1998 | 9923739 |
| a novel expression system based on host-range expansion of baculovirus. | a host range expanded recombinant autographa californica multiple-nucleocapsid nucleopolyhedrosis virus acmnpv/r2 was obtained by cotransfection of the bacmid dna from escherichia coli dh10bac along with a plasmid pbmh-m containing hindiii m fragment of bombyx mori nuclear polyhedrosis virus (bmnpv) genomic dna. a recombinant transposon vector carrying a mutant green fluorescent protein gene (gfp) and a polyhedrin gene was constructed. transposition was carried out in both e. coli dh10bac and e. ... | 1998 | 9923744 |
| concomitant primary infection of the midgut epithelial cells and the hemocytes of trichoplusia ni by autographa californica nucleopolyhedrovirus. | we have constructed a modified autographa californica multicapsid nucleopolyhedrovirus (acmnpv) to express the green fluorescent protein (gfp) under the polyhedrin promoter and used it to study the infection process of acmnpv in trichoplusia ni larvae. t. ni larvae that ingested the virus showed localized expression of gfp in the midgut epithelial cells and the hemocytes at 12 h post infection (hpi). the presence of gfp-related fluorescence in the midgut columnar cells indicated that the virus w ... | 1998 | 10036786 |
| optimising fed-batch production of recombinant proteins using the baculovirus expression vector system. | fed-batch culture can offer significant improvement in recombinant protein production compared to batch culture in the baculovirus expression vector system (bevs), as shown by nguyen et al. (1993) and bedard et al. (1994) among others. however, a thorough analysis of fed-batch culture to determine its limits in improving recombinant protein production over batch culture has yet to be performed. in this work, this issue is addressed by the optimisation of single-addition fed-batch culture. this t ... | 1998 | 10099329 |
| [the envelope formation of acmnpv and the location of envelope protein gp64 in host cell]. | electron microscopic study on the recombinant acmnpv infected sf9 cell showed that the virus has at least two forms of envelope formation: one is budding from plasma membrane and the other is enveloping in the nucleus by membrane structure. an additional possible pathway is budding from nuclear membrane. the envelope formation of acmnpv in sf9 cell line and its relationship with the envelope protein gp64 were investigated by using immunofluorescence techniques. the results showed gp64 was mainly ... | 1998 | 12549395 |
| characterization of the acidic domain of the ie1 regulatory protein from orgyia pseudotsugata multicapsid nucleopolyhedrovirus. | this study presents a detailed analysis of the acidic n-terminal region of the orgyia pseudotsugata multicapsid nucleopolyhedrovirus (opmnpv) transactivator ie1. the n-terminal region of ie1 is rich in acidic amino acids and has been hypothesized to be an acidic activation domain. removal of the n-terminal 126 amino acids containing the acidic domain of ie1 resulted in complete loss of transactivation activity, indicating that this region is essential for transactivation. the opmnpv acidic domai ... | 1998 | 9875318 |
| autographa californica nuclear polyhedrosis virus: subcellular localization and protein trafficking of bv/odv-e26 to intranuclear membranes and viral envelopes. | the autographa californica nuclear polyhedrosis virus da26 gene codes for an envelope protein of both budded virus (bv) and occlusion derived virus (odv). western blot and temporal analysis of infected cell extracts detected a protein of 26 kda by 4 h postinfection (p.i.). the amount of protein increased by 16 h p.i. and remained at high levels throughout infection. by 36 h p.i. several additional immunoreactive proteins were detected which migrated at approximately 18 kda and remained through 9 ... | 1998 | 9448690 |
| identification and characterization of a protein kinase-interacting protein encoded by the autographa californica nuclear polyhedrosis virus. | we have used a yeast two-hybrid system to identify a baculoviral gene encoding a protein kinase-interacting protein (pkip). the pkip gene is located at 15.8-16.2 m.u (map units) and represents orf 24 in the sequence of ayres et al. (1994) of the autographa californica nuclear polyhedrosis virus. pkip is a 19.2-kda protein and appears to stimulate the activity of the viral protein kinase-1 in vitro. northern blotting analysis revealed that pkip is a late gene. attempts to plaque purify a pkip- vi ... | 1998 | 9454690 |
| purification and kinetic analysis of a baculovirus ecdysteroid udp-glucosyltransferase. | the baculovirus ecdysteroid udp-glucosyltransferase (egt) disrupts the hormonal balance of the insect host by catalysing the conjugation of ecdysteroids, the moulting hormones, with the sugar moiety from udp-glucose or udp-galactose. in this study, autographa californica nucleopolyhedrovirus egt has been overproduced and purified, and its kinetic properties determined. the enzyme was purified 1100-fold to near-homogeneity using only two major steps, ion-exchange and gel-filtration chromatography ... | 1998 | 9494095 |
| lymantria dispar nucleopolyhedrovirus hrf-1 expands the larval host range of autographa californica nucleopolyhedrovirus. | the gypsy moth (lymantria dispar) is nonpermissive for autographa californica nucleopolyhedrovirus (acnpv) infection. we previously isolated a gene, host range factor 1 (hrf-1), from l. dispar nucleopolyhedrovirus that promotes acnpv replication in ld652y cells, a nonpermissive l. dispar cell line (s. m. thiem, x. du, m. e. quentin, and m. m. berner, j. virol. 70:2221-2229, 1996). in the present study, we investigated the ability of hrf-1 to alter the larval host range of acnpv. bioassays using ... | 1998 | 9499118 |
| temporal expression of the acmnpv lef-4 gene and subcellular localization of the protein. | autographa californica nucleopolyhedrovirus (acmnpv) lef-4 gene [orf 90; ayres et al. (1994) virology 202, 586-605] is involved in both late and very late gene expression [passarelli and miller (1993) virology 197, 704-714]. the transcriptional properties of this gene have been analyzed. it is transcribed as a single 1.6-kb mrna and transcripts were first detected 3 h postinfection (pi). the extremities of the transcript have been mapped by primer extension and 3' race-pcr to positions -56 from ... | 1998 | 9499802 |
| two key mutations in the host-range specificity domain of the p143 gene of autographa californica nucleopolyhedrovirus are required to kill bombyx mori larvae. | autographa californica nucleopolyhedrovirus (acmnpv) does not replicate in bombyx mori cells (bm5, bmn). we have shown previously that when a short dna sequence within acmnpv orf95, which encodes the viral helicase p143, is replaced with the colinear region of b. mori nucleopolyhedrovirus (bmnpv), acmnpv gains the ability to replicate in bm5 cells. to determine the mutational events in the p143 gene required to allow acmnpv replication in b. mori cells, acmnpv recombinants produced in sf9 cells ... | 1998 | 9568990 |
| autographa californica nucleopolyhedrovirus infection results in sf9 cell cycle arrest at g2/m phase. | baculovirus infection results in the induction of membrane structures within the nucleoplasm of the host cells. the source of these membranes is unclear; however, using the normal dynamics of cellular membranes and the nuclear envelope as a model, it is possible that the cell cycle might play a role in the regulation of formation of these intranuclear membranes. therefore, one goal of this study was to investigate the effect of baculovirus infection on the cell cycle of sf9 host cells. since few ... | 1998 | 9581791 |
| mapping of orf121, a factor that activates baculovirus early gene expression. | the protein product of the 39k gene of autographa californica nuclear polyhedrosis virus is thought to be important for viral replication because of its association with the virogenic stroma and its role in activation of late gene expression transient expression assays showed that addition of a dna fragment encoding a 58-amino-acid polypeptide increased expression of a 39k reporter plasmid. this stimulation was dependent on cotransfection of a plasmid encoding ie1. cotransfection of this gene, o ... | 1998 | 9601517 |
| homologous p35 proteins of baculoviruses show distinctive anti-apoptotic activities which correlate with the apoptosis-inducing activity of each virus. | the anti-apoptotic activity of p35s from two baculoviruses, autographa californica nucleopolyhedrovirus (acnpv) and bombyx mori npv (bmnpv), was compared in mammalian cells. acnpv p35 efficiently blocked apoptosis induced by caspase overexpression, but bmnpv p35 did so very poorly. analysis of chimeric p35s and in vitro cleavage of wild type p35s suggest that the cleavage efficiency of p35 correlates with the blocking activity. single amino acid substitutions of bmnpv p35 with those observed in ... | 1998 | 9613616 |
| expression and mutational analysis of the baculovirus very late factor 1 (vlf-1) gene. | we have examined the expression and function of a gene, vlf-1, of autographa californica nuclear polyhedrosis virus that is known to encode a regulator of very late gene transcription. western blot analysis revealed that vlf-1 is expressed during the late phase of infection, primarily from 15 to 24 h postinfection. vlf-1 localized in the cell nucleus and was also present in the nucleocapsids of virus particles. mapping of vlf-1 mrna by primer extension showed that transcription initiates at a ta ... | 1998 | 9614871 |
| effects of substituting granulin or a granulin-polyhedrin chimera for polyhedrin on virion occlusion and polyhedral morphology in autographa californica multinucleocapsid nuclear polyhedrosis virus. | substitution of granulin from the trichoplusia ni granulosis virus (tngv) for polyhedrin of the autographa californica multinucleocapsid nuclear polyhedrosis virus (acmnpv) yielded a few very large (2 to 5 micron) cuboidal inclusions in the cytoplasm and nucleus of infected cells. these polyhedra lacked the beveled edges characteristic of wild-type acmnpv polyhedra, contained fractures, and occluded few virions. placing a nuclear localization signal (krkk) in granulin directed more granulin to t ... | 1998 | 9621097 |
| the ecdysteroid udp-glucosyltransferase gene of autographa californica nucleopolyhedrovirus alters the moulting and metamorphosis of a non-target insect, the silkworm, bombyx mori (lepidoptera, bombycidae). | the autographa californica nucleopolyhedrovirus (acmnpv) does not infect the silkworm and molecular studies on silkworm insusceptibility have not been performed. in cultured cells of the silkworm, the expression of viral genes has been reported. the expression of acmnpv genes and their effect in vivo and in vitro was studied. in this study, the early gene, the ecdysteroid udp-glucosyltransferase (egt) gene of acmnpv, which inactivates the insect moulting hormone by sugar conjugation, was examine ... | 1998 | 9634100 |
| the single-nucleocapsid nucleopolyhedrovirus of buzura suppressaria encodes a p10 protein. | the p10 gene of buzura suppressaria single-nucleocapsid nucleopolyhedrovirus (busunpv) was identified by virtue of its localization downstream from the autographa californica (ac) mnpv p26 homologue. the busunpv p10 gene encodes a protein of 94 amino acids. the amino acid sequence contains domains characteristic of baculovirus p10 proteins, e.g. a coiled-coil domain, a proline-rich motif and a positively charged c terminus. the highest amino acid homologies were found with the spodoptera littora ... | 1998 | 9634101 |
| observations on the presence of the peritrophic membrane in larval trichoplusia ni and its role in limiting baculovirus infection. | light microscopical examinations of dissected and stained peritrophic membranes (pms) were conducted to determine the presence or absence of this protective structure in larvae of trichoplusia ni, prior to and through ecdysis. observations of fourth- and fifth-instar larvae of t. ni from two independent rearing colonies showed that pms were present and lined the midgut prior to, during, and immediately after ecdysis in both instars. western blot analysis of insect intestinal mucin (iim), a major ... | 1998 | 9647702 |
| a baculovirus mutant defective in pkip, a protein which interacts with a virus-encoded protein kinase. | we have found that a temperature-sensitive mutant of the baculovirus acmnpv, tsb97, is defective in pkip, the product of orf24 which was previously found to interact with and stimulate the activity of a virus-encoded protein kinase, pk-1. the mutant lacks the ability to form plaques and occlusion bodies at the nonpermissive temperature. the mutant displays several properties which suggest a defect in the latter half of the late phase of infection; these properties include a delay in the shutoff ... | 1998 | 9657956 |
| expression of feline recombinant interferon-gamma in baculovirus and demonstration of biological activity. | we have previously reported the cloning of the coding sequence for feline-specific interferon-gamma. here, we describe the expression of this sequence in a baculovirus system and demonstrate the biological activity of the recombinant protein. the coding sequence for feline interferon was directionally cloned into the baculovirus transfer vector paccl29-1. transfer vector and linearized wild-type acmnpv (bacpak6) were used to co-transfect sf9 cells by calcium phosphate coprecipitation. subsequent ... | 1998 | 9661260 |
| a baculovirus single-stranded dna binding protein, lef-3, mediates the nuclear localization of the putative helicase p143. | the intracellular localization of the baculovirus autographa californica nuclear polyhedrosis virus p143 gene product (p143) was investigated by immunofluoresence staining of infected and transfected cells. as expected for a protein essential for viral dna replication, under these conditions, p143 was localized to the nucleus. however, when a plasmid directing the synthesis of p143 from its own promoter was co-transfected with a plasmid expressing ie-1, p143 was found in the cytoplasm. cotransfe ... | 1998 | 9683569 |
| characterization of a baculovirus-encoded rna 5'-triphosphatase. | autographa californica nuclear polyhedrosis virus (acnpv) encodes a 168-amino-acid polypeptide that contains the signature motif of the superfamily of protein phosphatases that act via a covalent cysteinyl phosphate intermediate. the sequence of the acnpv phosphatase is similar to that of the rna triphosphatase domain of the metazoan cellular mrna capping enzyme. here, we show that the purified recombinant acnpv protein is an rna 5'-triphosphatase that hydrolyzes the gamma-phosphate of triphosph ... | 1998 | 9696798 |
| construction of occluded recombinant baculoviruses containing the full-length cry1ab and cry1ac genes from bacillus thuringiensis. | the administration of baculoviruses to insects for bioassay purposes is carried out, in most cases, by contamination of food surfaces with a known amount of occlusion bodies (obs). sinceper os infection is the natural route of infection, occluded recombinant viruses containing crystal protein genes (cry1ab and cry1ac) from bacillus thuringiensis were constructed for comparison with the baculovirus prototype autographa californica nucleopolyhedrovirus (acnpv). the transfer vector pacuw2b was used ... | 1998 | 9698820 |
| control of baculovirus polyhedrin gene expression by very late factor 1. | vlf-1 is a baculovirus gene that regulates very late gene expression (j. r. mclachlin and l. k. miller, j. virol., 68, 7746-7756, 1994) and also plays a crucial role in the replication of the budded form of autographa californica nuclear polyhedrosis virus (acmnpv) (s. yang and l. k. miller, "expression and mutational analysis of the baculovirus very late factor 1 (vlf-1) gene." virology, 245, 99-109, 1998). to examine the influence of vlf-1 expression on baculovirus infection, we constructed re ... | 1998 | 9705262 |
| a protein tyrosine phosphatase-like protein from baculovirus has rna 5'-triphosphatase and diphosphatase activities. | the superfamily of protein tyrosine phosphatases (ptps) includes at least one enzyme with an rna substrate. we recently showed that the rna triphosphatase domain of the caenorhabditis elegans mrna capping enzyme is related to the ptp enzyme family by sequence similarity and mechanism. the ptp most similar in sequence to the capping enzyme triphosphatase is bvp, a dual-specificity ptp encoded by the autographa californica nuclear polyhedrosis virus. although bvp previously has been shown to have ... | 1998 | 9707557 |
| a highly conserved genomic region in baculoviruses: sequence analysis of an 11.3 kbp dna fragment (46.5-55.1 m.u.) of the spodoptera exigua multicapsid nucleopolyhedrovirus. | a dna fragment of 11.3 kilobase pairs (kbp) in size of the baculovirus spodoptera exigua multicapsid nucleopolyhedrovirus (semnpv) genome (46.5 to 55.1 m.u.) was completely sequenced. analysis of the sequence revealed eleven potential open reading frames (orf). ten of these orfs showed significant amino acid identity to autographa californica mnpv (acmnpv) and orgyia pseudotsugata mnpv (opmnpv) genes p6.9, lef5, 38k, p19, p143, p25, p18, vp33, lef4, and vp39. one orf (xc12) has no homolog in oth ... | 1998 | 9725671 |
| a virus-encoded rna polymerase purified from baculovirus-infected cells. | a dna-dependent rna polymerase was purified to homogeneity, starting from insect cells infected with the baculovirus autographa californica nuclear polyhedrosis virus (acnpv). the purified polymerase supported accurate and specific transcription from late and very late promoters but was not active on viral early promoters. thus, promoter recognition is an integral function of the purified enzyme. the purified rna polymerase was composed of only four equimolar subunits, which makes it the simples ... | 1998 | 9733837 |
| the structural protein odv-ec27 of autographa californica nucleopolyhedrovirus is a multifunctional viral cyclin. | two major characteristics of baculovirus infection are arrest of the host cell at g2/m phase of the cell cycle with continuing viral dna replication. we show that autographa californica nucleopolyhedrovirus (acmnpv) encodes for a multifunctional cyclin that may partially explain the molecular basis of these important characteristics of acmnpv (baculovirus) infection. amino acids 80-110 of the viral structural protein odv-ec27 (-ec27) demonstrate 25-30% similarity with cellular cyclins within the ... | 1998 | 9736714 |
| identification and characterization of the cydia pomonella granulovirus cathepsin and chitinase genes. | a 3.2 kb bamhi-ecori fragment of the cydia pomonella granulovirus (cpgv) genome was subcloned and characterized. sequence analysis revealed two complete and one partial open reading frames (orfs). orf7l is predicted to encode a 66.7 kda protein (594 amino acid residues) that is 57% identical (amino acid sequence) to the chia gene (orf126) of autographa californica nucleopolyhedrovirus (acmnpv), encoding a chitinase. orf8r is 333 amino acids in length and shows high similarity (between 64% and 67 ... | 1998 | 9747739 |
| persistent baculovirus infection results from deletion of the apoptotic suppressor gene p35. | infection with the wild-type baculovirus autographa californica multiple nuclear polyhedrosis virus (acmnpv) results in complete death of spodoptera frugiperda (sf) cells. however, infection of sf cells with acmnpv carrying a mutation or deletion of the apoptotic suppressor gene p35 allowed the cloning of surviving sf cells that harbored persistent viral genomes. persistent infection established with the virus with p35 mutated or deleted was blocked by stable transfection of p35 in the host geno ... | 1998 | 9765462 |
| the impact of host developmental status on baculovirus replication | the yield of progeny occlusion bodies from a baculovirus infection is a critical parameter governing viral population dynamics in the field. previous evidence has suggested that the ability of the virus to block host development may be an important factor in determining yield of progeny virus. here, we explore the relationship between yield, dose, and host developmental status at the time of infection during autographa californica nucleopolyhedrovirus infection of final instar heliothis virescen ... | 1998 | 9784350 |
| new cell lines from heliothis virescens: characterization and susceptibility to baculoviruses | new cell lines from embryos of heliothis virescens were recently developed. six primary cultures were initiated in june 1995. from these initial cultures, two produced sufficient cell growth to allow subcultivation and eventually led to the establishment of seven cell strains, three of which are maintained at low temperatures (17 degreesc). the strains were compared with a previously established cell line from h. virescens by isozyme analysis and shown to be from the same species. all the strain ... | 1998 | 9784351 |
| the early baculovirus he65 promoter: on the mechanism of transcriptional activation by ie1. | we have initiated studies on the mechanism of early transcriptional activation of the early he65 promoter during infection with autographa californica multicapsid nuclear polyhedrosis virus. this analysis is based on a comparison of the sequences required for he65 promoter activation with those sequences that support specific protein binding. the he65 promoter is located immediately downstream of the homologous region (hr) 4a. the sequences of hr4a are characterized by two imperfect palindromes ... | 1998 | 9791025 |
| mutational analysis of the n-linked glycans on autographa californica nucleopolyhedrovirus gp64. | gp64 is the major envelope glycoprotein in the budded form of autographa californica multicapsid nucleopolyhedrovirus (acmnpv). gp64 is essential for acmnpv infection, as it mediates penetration of budded virus into host cells via the endocytic pathway. in this study, we used site-directed mutagenesis to map the positions of the n-linked glycans on acmnpv gp64, characterize their structures, and evaluate their influence on gp64 function. we found that four of the five consensus n-glycosylation s ... | 1998 | 9811679 |
| guanylyltransferase activity of the lef-4 subunit of baculovirus rna polymerase. | the baculovirus autographa californica nuclear polyhedrosis virus encodes a dna-dependent rna polymerase that transcribes viral late genes. this polymerase is composed of four equimolar subunits, lef-4, lef-8, lef-9, and p47. here we present data indicating that the lef-4 subunit of rna polymerase is a guanylyltransferase. incubation of rna polymerase in the presence of divalent cation and radiolabeled gtp resulted in the formation of a covalent enzyme-guanylate complex that comigrated with the ... | 1998 | 9811738 |
| the lef-4 subunit of baculovirus rna polymerase has rna 5'-triphosphatase and atpase activities. | the baculovirus autographa californica nuclear polyhedrosis virus encodes a dna-dependent rna polymerase that is required for transcription of viral late genes. this polymerase is composed of four equimolar subunits, lef-8, lef-4, lef-9, and p47. the lef-4 subunit has guanylyltransferase activity, suggesting that baculoviruses may encode a full complement of capping enzymes. here we show that lef-4 is a bifunctional enzyme that hydrolyzes the gamma phosphates of triphosphate-terminated rna and a ... | 1998 | 9811739 |
| rna 5'-triphosphatase, nucleoside triphosphatase, and guanylyltransferase activities of baculovirus lef-4 protein. | autographa californica nuclear polyhedrosis virus late and very late mrnas are transcribed by an rna polymerase consisting of four virus-encoded polypeptides: lef-8, lef-9, lef-4, and p47. the 464-amino-acid lef-4 subunit contains the signature motifs of gtp:rna guanylyltransferases (capping enzymes). here, we show that the purified recombinant lef-4 protein catalyzes two reactions involved in rna cap formation. lef-4 is an rna 5'-triphosphatase that hydrolyzes the gamma phosphate of triphosphat ... | 1998 | 9811740 |
| nineteen baculovirus open reading frames, including lef-12, support late gene expression. | a set of 18 plasmid subclones of the autographa californica nuclear polyhedrosis virus genome, each containing an identified late expression factor gene (lef), supports expression from a late viral promoter in transient expression assays in the sf-21 cell line derived from spodoptera frugiperda. we have constructed a further set of plasmids in which each lef open reading frame (orf) is controlled by the drosophila melanogaster heat shock protein 70 (hsp70) promoter and epitope tagged. failure of ... | 1998 | 9811761 |
| localization of a baculovirus-induced chitinase in the insect cell endoplasmic reticulum. | confocal immunofluorescence microscopy was used to demonstrate that the autographa californica nucleopolyhedrovirus (acmnpv) chitinase was localized within the endoplasmic reticulum (er) of virus-infected insect cells. this was consistent with removal of the signal peptide from the chitinase and an er localization motif (kdel) at the carboxyl end of the protein. chitinase release from cells, a prerequisite for liquefaction of virus-infected insect larvae, appears to be aided by synthesis of the ... | 1998 | 9811762 |
| coinfection of spodoptera exigua and spodoptera frugiperda cell lines with the nuclear polyhedrosis viruses of autographa californica and spodoptera exigua. | the autographa californica multinucleocapsid nuclear polyhedrosis virus (acmnpv) has a broad host range among lepidoptera. in contrast, the spodoptera exigua mnpv (semnpv) can replicate efficiently only in s. exigua larvae or s. exigua-derived cell lines. in this study, we examined the coinfection of s. exigua se301 and spodoptera frugiperda iplb-sf21aeii (sf21) cell lines with semnpv and acmnpv recombinant (ac360-501beta-gal) which was constructed for expression of beta-galactosidase under cont ... | 1998 | 10325534 |
| role of baculovirus ie2 and its ring finger in cell cycle arrest. | the ie2 gene of autographa californica nuclear polyhedrosis virus (acmnpv) is known to transactivate transient expression from viral promoters in a host cell-specific manner. we report that transfection of spodoptera frugiperda (sf-21) cells with ie2 was sufficient to arrest the cell cycle, resulting in the accumulation of enlarged cells with abnormally high dna contents. by 72 h posttransfection, more than 50% of ie2-transfected cells had dna contents greater than 4n. there was no evidence of m ... | 1998 | 9420274 |
| the pnk/pnl gene (orf 86) of autographa californica nucleopolyhedrovirus is a non-essential, immediate early gene. | autographa californica nucleopolyhedrovirus (acmnpv) orf 86, located within the hindiii c fragment, potentially encodes a protein which shares sequence similarity with two t4 bacteriophage gene products, rna ligase and polynucleotide kinase. this acmnpv gene has been designated pnk/pnl but has yet to be assigned a function in virus replication. it has been classified as an immediate early virus gene, since the promoter was active in uninfected insect cells and mrna transcripts were detectable fr ... | 1998 | 9519844 |
| in vitro transcription of pe38/polyhedrin hybrid promoters reveals sequences essential for recognition by the baculovirus-induced rna polymerase and for the strength of very late viral promoters. | in vitro transcription was used to analyze the promoter specificity of the alpha-amanitin-resistant rna polymerase that is induced late during infection of autographa californica multicapsid nuclear polyhedrosis virus. by modifying the preparation of crude nuclear extracts, we have established an assay that permits differentiation between weak late and strong very late viral promoters. the virus-induced rna polymerase initiates at a taag sequence motif in both late and very late promoters. based ... | 1998 | 9525621 |
| studies of the nucleopolyhedrovirus infection process in insects by using the green fluorescence protein as a reporter. | a recombinant autographa californica multicapsid nucleopolyhedrovirus (acmnpv) expressing the green fluorescence protein (gfp) under the control of the acmnpv polyhedrin promoter was constructed to study the spatial and temporal regulation of baculovirus infection in a permissive host. larvae that ingested acmnpv-gfp showed localized expression of gfp in the midgut epithelial cells, as well as hemocytes, at 24 h postinfection. the presence of fluorescence in these tissues indicated not only that ... | 1998 | 9525665 |
| species-specificity in endoplasmic reticulum signal peptide utilization revealed by proteins from trypanosoma brucei and leishmania. | n-terminal signal peptides direct secretory and most membrane proteins into the exocytic pathway at the endoplasmic reticulum. signal sequences can function across kingdoms. however, our attempts at translocating variant surface glycoprotein (vsg) 117, vsg mvat7, vsg 221 and bip from trypanosoma brucei and gp63 from leishmania chagasi into canine pancreas microsomes failed. on replacing the signal peptide of vsg 117 with that from yeast prepro-alpha-mating factor (ppalphamf) the chimaeric protei ... | 1998 | 9531493 |
| development and validation of a competitive enzyme-linked immunosorbent assay for detection of type a influenza antibodies in avian sera. | serologic screening of avian sera for group-specific antibodies to type a influenza is currently accomplished by using the avian influenza (ai) agar gel immunodiffusion (agid) test. a competitive enzyme-linked immunosorbent assay (celisa) was developed using a baculovirus vector, autographa californica nuclear polyhedrosis virus, expressing the nucleoprotein (np) gene of a/ann arbor/6/60 influenza virus. the recombinant np was obtained by inoculation of spodoptera frugiperda (sf9) insect cells o ... | 1998 | 9533078 |
| in vitro and in vivo host range of anticarsia gemmatalis multiple nuclear polyhedrosis virus. | a clone of the wild type (wt) anticarsia gemmatalis multiple nuclear polyhedrosis virus agmnpv, derived from a geographical isolate (hondrina, brazil) and designated agmnpv-cl4-3a1, was used to determine the host range of this virus in six established lepidopteran cell lines: anticarsia gemmatalis (bcirl-ag-am1), helicoverpa zea (bcirl-hz-am1), heliothis virescens (bcirl-hv-am1), helicoverpa armigera (bcirl-ha-am1), trichoplusia ni (tn-cl1), bombyx mori (bmn), and a coleopteran cell line anthono ... | 1998 | 9542640 |
| removal of 106 amino acids from the n-terminus of udp-glcnac: alpha-3-d-mannoside beta-1,2-n-acetylglucosaminyltransferase i does not inactivate the enzyme. | udp-glcnac: alpha-3-d-mannoside beta-1,2-n-acetylglucosaminyltransferase i (gnti, ec 2.4.1.101) plays an essential role in the conversion of oligomannose to complex and hybrid n-glycans. rabbit gnti is 447 residues long and has a short four-residue n-terminal cytoplasmic tail, a 25-residue putative signal-anchor hydrophobic domain, a stem region of undetermined length and a large c-terminal catalytic domain, a structure typical of all glycosyltransferases cloned to date. comparison of the amino ... | 1998 | 9557881 |
| cell-cycle perturbation in sf9 cells infected with autographa californica nucleopolyhedrovirus. | flow cytometry analysis of the cell-cycle progression was performed in sf9 cells infected with autographa californica nucleopolyhedrovirus (acnpv) in the cultures partially synchronized by aphidicolin exposure and deprivation. cells infected with acnpv during the g1 phase progressed and were arrested in the s phase in the 4 h following the infection, whereas cells infected during the s phase did not progress past the s phase. cells infected during the g2/m phase remained in the g2/m phase withou ... | 1999 | 10329579 |
| human pir1 of the protein-tyrosine phosphatase superfamily has rna 5'-triphosphatase and diphosphatase activities. | a human cdna was isolated encoding a protein with significant sequence similarity (41% identity) to the bvp rna 5'-phosphatase from the autographa californica nuclear polyhedrosis virus. this protein is a member of the protein-tyrosine phosphatase (ptp) superfamily and is identical to pir1, shown by yuan et al. (yuan, y., da-ming, l., and sun, h. (1998) j. biol. chem. 272, 20347-20353) to be a nuclear protein that can associate with rna or ribonucleoprotein complexes. we demonstrate that pir1 re ... | 1999 | 10347225 |
| temperature dependent characteristics of a recombinant infectious hematopoietic necrosis virus glycoprotein produced in insect cells. | a recombinant infectious hematopoietic necrosis virus (ihnv) glycoprotein (g protein) was produced in insect cells using a baculovirus vector (autographa californica nuclear polyhedrosis virus). characteristics of this protein were evaluated in relation to native viral g protein. a full-length (1.6 kb) cdna copy of the glycoprotein gene of ihnv was inserted into the baculovirus vector under control of the polyhedrin promoter. high levels of g protein (approximately 0.5 microgram/1 x 10(5) cells) ... | 1999 | 10349547 |
| generation of a new protein purification matrix by loading ceramic hydroxyapatite with metal ions--demonstration with poly-histidine tagged green fluorescent protein. | the gene encoding the green fluorescent protein (gfp) from the jellyfish aequorea victoria, was inserted under transcriptional control of the polyhedrin promoter of the autographa californica nuclear polyhedrosis virus and expressed in the spodoptera frugiperda insect cell line sf9 during viral infection. the baculovirus transfervector pbluebachisb was used for constructing the recombinant baculovirus, so that the green fluorescent protein could be tagged with a poly-histidine tail. this fusion ... | 1999 | 10361722 |
| host cell receptor binding by baculovirus gp64 and kinetics of virion entry. | gp64 is the major envelope glycoprotein from budded virions of the baculoviruses autographa californica multicapsid nucleopolyhedrovirus (acmnpv) and orgyia pseudotsugata multicapsid nucleopolyhedrovirus (opmnpv). to examine the potential role of gp64 as a viral attachment protein in host cell receptor binding, we generated, overexpressed, and characterized a soluble form of the opmnpv gp64 protein, gp64solop. assays for trimerization, sensitivity to proteinase k, and reduction by dithiothreitol ... | 1999 | 10366584 |
| mitochondrial dna acts as potential promoter of the baculovirus rna polymerase. | we have examined whether mitochondrial dna could act as target of the rna polymerase encoded by the baculovirus autographa californica multicapsid nuclear polyhedrosis virus, because the baculovirus late promoters and the control region of host mitochondrial dna show a high degree of sequence similarity. in vitro transcription using mitochondrial dna from spodoptera frugiperda cells and nuclear extracts prepared from baculovirus infected cells demonstrates that mitochondrial dna is recognized by ... | 1999 | 10384964 |
| global protein synthesis shutdown in autographa californica nucleopolyhedrovirus-infected ld652y cells is rescued by trna from uninfected cells. | global protein synthesis arrest occurs in autographa californica nucleopolyhedrovirus (acnpv)-infected ld652y cells at late times postinfection (p.i.). a lymantria dispar nucleopolyhedrovirus gene, hrf-1, precludes this protein synthesis arrest. we used in vitro translation assays to characterize the translation defect. cell-free lysates prepared from uninfected ld652y cells, acnpv-infected cells harvested at early times p.i., and cells infected with vachrf-1, a recombinant acnpv bearing hrf-1, ... | 1999 | 10417257 |
| host-specific transcription of nucleopolyhedrovirus gene homologues in productive and abortive anticarsia gemmatalis mnpv infections. | in a previous report, we showed that anticarsia gemmatalis nucleopolyhedrovirus (agmnpv) infections of choristoneura fumiferana ipri-cf-124t and bombyx mori bm-5 cell lines are abortive, whereas a. gemmatalis ufl-ag-286 cells efficiently produce infectious virus and polyhedral inclusion bodies (pibs). in the present study, we explored transcription patterns in these infections using representative temporal classes of autographa californica mnpv (acmnpv) genes. northern analyses were carried out ... | 1999 | 10446647 |
| role of the 3' untranslated region of baculovirus p10 mrna in high-level expression of foreign genes. | the p10 gene of autographa californica nucleopolyhedrovirus has two putative aataaa polyadenylation signals. the downstream signal is used predominantly, as was determined by analysing 3' cdna ends. this downstream motif is followed by a gt-rich sequence, known to be important for efficient polyadenylation in mammalian systems. to analyse the importance of polyadenylation for p10 gene expression, recombinant viruses with altered 3' untranslated regions (utrs) were tested using chloramphenicol ac ... | 1999 | 10466825 |
| the establishment of heliothine cell lines and their susceptibility to two baculoviruses. | a total of eight cell lines were established from helicoverpa armigera (3) and h. punctigera (5) embryos and ovaries. cell lines were established and grown in tc100 and/or tc199-mk containing 10% fetal bovine serum. the serum-free medlium excell 400 was also used, with and without 10% supplemental fetal bovine serum, but failed to generate cell lines from fat bodies, embryos, or ovarian tissues. cell lines consisted of heterogenous cell types ranging from oval to fibroblast-like. this is the fir ... | 1999 | 10475263 |
| comparison of cell line maintenance procedures on insect cells used for producing baculoviruses. | a gypsy moth cell line, iplb-ldeita, maintained under various conditions was tested for susceptibility to and productivity of two baculoviruses, the autographa californica nucleopolyhedrovirus (acmnpv) and lymantria dispar nucleopolyhedrovirus (ldmnpv). the results suggest that cells maintained in serum-containing medium (modified tc100) were more susceptible (on the basis of titers in an endpoint assay) to ldmnpv than cells maintained in a serum-free medium (excell 400). such a difference was n ... | 1999 | 10475270 |
| the establishment of new cell lines from pseudaletia unipuncta with differential responses to baculovirus infection. | six insect cell lines from pseudaletia unipuncta embryos were established and characterized, and their susceptibility to autographa californica multiple nuclear polyhedrosis virus (acmnpv) infection was investigated. these embryonic p. unipuncta cell lines had characteristics distinct from each other in morphology and growth, and showed differential responses to acmnpv infection. among the six cell lines, two were highly susceptible to virus infection. one of these two cell lines, bti-pu-a7s, pr ... | 1999 | 10476920 |
| mutations within the autographa californica nucleopolyhedrovirus fp25k gene decrease the accumulation of odv-e66 and alter its intranuclear transport. | previous reports indicate that mutations within the autographa californica nucleopolyhedrosis virus fp25k gene (open reading frame 61) significantly reduce incorporation of enveloped nucleocapsids into viral occlusions. we report that fp25k is a nucleocapsid protein of both the budded virus (bv) and occluded virus (odv), and we describe the effects of two fp25k mutations (480-1 [n-terminal truncation] and fp-betagal [c-terminal fusion]) on the expression and cellular localization of odv-e66 and ... | 1999 | 10482609 |
| the characterization and phylogenetic relationship of the trichoplusia ni single capsid nuclear polyhedrosis virus polyhedrin gene. | the polyhedrin gene (polh) was identified from the trichoplusia ni (tni) single capsid nuclear polyhedrosis virus (snpv). an ecori fragment containing the truncated polyhedrin gene was detected by hybridization with an acmnpv expression vector probe; the remaining portion of the gene was amplified by reverse pcr. an open reading frame (orf) of 741 nucleotides (nt), encoding a putative protein of 246 amino acids (a.a) with mr 28,780 da was identified. the 5'-noncoding region contained the putativ ... | 1999 | 10499452 |
| purification of autographa californica nucleopolyhedrovirus dna polymerase from infected insect cells. | autographa californica nucleopolyhedrovirus (acmnpv) dna polymerase was purified from virus-infected cells using conventional chromatographic methods. the enzymatic activity of fractions eluting from single-stranded agarose gels was found to exactly coincide with a single polypeptide with an apparent molecular mass of approximately 110,000 da on denaturing polyacrylamide gels stained with coomassie blue. this purification scheme resulted in a 228-fold purification of acmnpv dna polymerase with r ... | 1999 | 10501509 |
| sequence analysis of the xestia c-nigrum granulovirus genome. | the nucleotide sequence of the xestia c-nigrum granulovirus (xcgv) genome was determined and found to comprise 178,733 bases with a g+c content of 40.7%. it contained 181 putative genes of 150 nucleotides or greater that showed minimal overlap. eighty-four of these putative genes, which collectively accounted for 43% of the genome, are homologs of genes previously identified in the autographa californica multinucleocapsid nucleopolyhedrovirus (acmnpv) genome. these homologs showed on average 33% ... | 1999 | 10502508 |
| expression, purification and characterization of peanut (arachis hypogaea) agglutinin (pna) from baculovirus infected insect cells. | peanut (arachis hypogaea) seed lectin, pna is widely used to identify tumor specific antigen (t-antigen), galbeta1-3galnac on the eukaryotic cell surface. the functional amino acid coding region of a cdna clone, pbsh-pn was pcr amplified and cloned downstream of the polyhedrin promoter in the autographa californica nucleopolyhedrovirus (acnpv) based transfer vector pvl1393. co-transfection of spodoptera frugiperda cells (sf9) with the transfer vector, pacpna and acrp6 (a recombinant acnpv having ... | 1999 | 10513900 |
| detection of baculovirus-infected insect cells by flow cytometric side-scatter analyses. | the baculovirus expression vector system (bevs), utilizing the autographa californica nuclear polyhedrosis virus (acnpv), has turned out to be an attractive alternative for high-level expression (<600 mg/l) of recombinant proteins. however, there is a shortage of reliable methods for monitoring the infection process in situations where marker proteins cannot be used. | 1999 | 10520205 |
| characterization of a nucleopolyhedrovirus from the black cutworm, agrotis ipsilon (lepidoptera: noctuidae) | the black cutworm, agrotis ipsilon (hufnagel) (lepidoptera: noctuidae), is a serious localized pest of vegetable and field crops. we have characterized a newly discovered baculovirus, the agrotis ipsilon multicapsid nucleopolyhedrovirus (agipmnpv), that was isolated from a. ipsilon in illinois. restriction enzyme fragment profiles of agipmnpv dna were distinct from those of previously described nucleopolyhedroviruses. electron microscopy of agipmnpv-infected tissues indicated that nucleocapsids ... | 1999 | 10534417 |
| expression of genes coding for animal virus glycoproteins in heterologous systems. | the outermost layers of animal viruses are usually composed of glycoproteins. they are responsible not only for the entrance of viruses into, and release from host cells but also for the initial interaction of a viral particle with immunological defense of the host. it is therefore not surprising that many laboratories devote a lot of effort to study viral glycoproteins at the molecular level. very often such studies are possible only after the introduction of a glycoprotein gene into a heterolo ... | 1999 | 10547034 |
| baculovirus stimulates antiviral effects in mammalian cells. | herein, we report that autographa californica nucleopolyhedrovirus, a member of the baculoviridae family, is capable of stimulating antiviral activity in mammalian cells. baculoviruses are not pathogenic to mammalian cells. nevertheless, live baculovirus is shown here to induce interferons (ifn) from murine and human cell lines and induces in vivo protection of mice from encephalomyocarditis virus infection. monoclonal antibodies specific for the baculovirus envelope gp67 neutralize baculovirus- ... | 1999 | 10559307 |
| sequence and organization of the spodoptera exigua multicapsid nucleopolyhedrovirus genome. | the nucleotide sequence of the dna genome of spodoptera exigua multicapsid nucleopolyhedrovirus (semnpv), a group ii npv, was determined and analysed. the genome contains 135611 bp and has a g+c content of 44 mol%. computer-assisted analysis revealed 139 orfs of 150 nucleotides or larger; 103 have homologues in autographa californica mnpv (acmnpv) and a further 16 have homologues in other baculoviruses. twenty orfs are unique to semnpv. major differences in semnpv gene content and arrangement we ... | 1999 | 10567663 |
| the nucleopolyhedroviruses of rachiplusia ou and anagrapha falcifera are isolates of the same virus. | the 7.8 kb ecori-g fragment of rachiplusia ou multicapsid nucleopolyhedrovirus (romnpv), containing the polyhedrin gene, was cloned and sequenced. the sequence of the fragment was 92.3% identical to the sequence of the corresponding region in the autographa californica (ac)mnpv genome. a comparison of the ecori-g sequence with other mnpv sequences revealed that romnpv was most closely related to acmnpv. however, the predicted amino acid sequence of romnpv polyhedrin shared more sequence identity ... | 1999 | 10573177 |
| baculovirus-based genetic screen for antiapoptotic genes identifies a novel iap. | the prototype baculovirus, autographa californica multiple nuclear polyhedrosis virus (acmnpv) expresses p35, a potent anti cell-death gene that promotes the propagation of the virus by blocking host cell apoptosis. infection of insect sf-21 cells with acmnpv lacking p35 induces apoptosis. we have used this pro-apoptotic property of the p35 null virus to screen for genes encoding inhibitors of apoptosis that rescue cells infected with the p35 defective virus. we report here the identification of ... | 1999 | 10593985 |
| development of a novel expression vector system using spodoptera exigua nucleopolyhedrovirus. | to develop a novel spodoptera exigua nucleopolyhedrovirus (senpv) expression vector system, we examined characteristics of the senpv polyhedrin expression in s. exigua cells (se301). while the extracellular virus titer of senpv was 100-fold lower than that of autographa californica nucleopolyhedrovirus (acnpv), the levels of polyhedral inclusion body (pib) formation and polyhedrin expression were higher in senpv. to investigate foreign gene expression under the control of the polyhedrin promoter ... | 1999 | 10597039 |
| mediated pathogenicity of the baculovirus acmnpv by the venom from euplectrus comstockii howard (hymenoptera: eulophidae). | the compatibility of the venom from the parasitic species euplectrus comstockii howard (hymenoptera: eulophidae) with the pathogenicity of autographa californica (speyer) (lepidoptera: noctuidae) mnpv baculovirus (acmnpv) was tested in third and fourth instar larvae of trichoplusia ni (hübner) (lepidoptera: noctuidae). the presence of acmnpv did not alter the ability of the venom to arrest ecdysis in t. ni larvae. the presence of the venom delayed the rate of viruses by acmnpv but increased the ... | 1999 | 10631800 |
| multiple nucleocapsid packaging of autographa californica nucleopolyhedrovirus accelerates the onset of systemic infection in trichoplusia ni. | among the nucleopolyhedroviruses (baculoviridae), the occlusion-derived virus (odv), which initiates infection in host insects, may contain only a single nucleocapsid per virion (the snpvs) or one to many nucleocapsids per virion (the mnpvs), but the significance of this difference is unclear. to gain insight into the biological relevance of these different packaging strategies, we compared pathogenesis induced by odv fractions enriched for multiple nucleocapsids (odv-m) or single nucleocapsids ... | 1999 | 9847346 |
| development and use of a pcr assay for detection of the reproductive virus in wild populations of helicoverpa zea (lepidoptera: noctuidae) | helicoverpa zea reproductive virus (hzrv) is a nonoccluded bacilliform virus that affects both female and male moths of the corn earworm h. zea. in order to study the biology and host range of hzrv, a bioassay was previously developed to detect the presence of this virus in infected insects. a drawback of this bioassay is that it is time consuming and requires more than a month to complete. here we describe the development of a polymerase chain reaction (pcr) assay for the rapid detection of hzr ... | 1999 | 9878296 |
| baculovirus p33 binds human p53 and enhances p53-mediated apoptosis. | in vertebrates, p53 participates in numerous biological processes including cell cycle regulation, apoptosis, differentiation, and oncogenic transformation. when insect sf-21 cells were infected with a recombinant of the baculovirus autographa californica nuclear polyhedrosis virus (acmnpv) overexpressing human p53, p53 formed a stable complex with the product of the acmnpv orf92, a novel protein p33. the interaction between p53 and p33 was further confirmed by immunoprecipitation studies. when ... | 1999 | 9882325 |
| isolation of an apoptosis suppressor gene of the spodoptera littoralis nucleopolyhedrovirus. | spodoptera frugiperda sf9 cells infected with mutants of the autographa californica nucleopolyhedrovirus (acmnpv) which lack a functional p35 gene undergo apoptosis, aborting the viral infection. the spodoptera littoralis nucleopolyhedrovirus (slnpv) was able to suppress apoptosis triggered by vdeltap35k/pol+, an acmnpv p35 null mutant. to identify the putative apoptotic suppressor gene of slnpv, overlapping cosmid clones representing the entire slnpv genome were individually cotransfected along ... | 1999 | 9882332 |
| a baculovirus enhancin alters the permeability of a mucosal midgut peritrophic matrix from lepidopteran larvae. | the peritrophic matrix (pm) in lepidopterous larvae may function as a defensive barrier against ingested viral pathogens. pms isolated from trichoplusia ni and pseudaletia unipuncta larvae, were treated with a baculovirus-encoded metalloprotease (enhancin) from trichoplusia ni granulosis virus (tngv) and their in vitro permeability to blue dextran and fluorescent-labelled autographa californica nuclear polyhedrosis virus (acmnpv) was determined using a dual chamber permeability apparatus. incuba ... | 1999 | 12770384 |
| efficient secretion of proteins expressed from insect cells directed by pam signal peptide. | signal and leading peptide sequences of rat pam was inserted into the baculovirus transfer vector, and secretion expression plasmids pbacpag2 and pbacpai for the fusion gene pabc-hgrf and pabc-igf-i were constructed, respectively. by cotransfection with linear genomic dna of modified autographa californica nuclear polyhedrosis virus (bacpak6) and homologous recombination, the recombinant acnpv, bacpag and bacpai, were obtained and identified. fusion proteins pabc-hgrf and pabc-igf-i were secrete ... | 1999 | 12142920 |
| human interleukin-2 production in insect (trichoplusia ni) larvae: effects and partial control of proteolysis. | many eukaryotic proteins have been successfully expressed in insect cells infected with a baculovirus in which the foreign gene has been placed under the control of a viral promoter. this system can be costly at large scale due to the quality of virus stock, problems of oxygen transfer, and severity of large-scale contamination. to circumvent this problem, we have investigated the expression of a foreign protein, human interleukin-2 (il-2), in insect larvae, trichoplusia ni, infected with the ba ... | 1999 | 10099527 |
| purification of human interleukin-2 fusion protein produced in insect larvae is facilitated by fusion with green fluorescent protein and metal affinity ligand. | the fusion protein of green fluorescent protein (gfp) and human interleukin-2 (hil-2) was produced in insect trichoplusia ni larvae infected with recombinant baculovirus derived from the autographa californica nuclear polyhedrosis virus (acnpv). this fusion protein was composed of a metal ion binding site (his)6 for rapid one-step purification using immobilized metal affinity chromatography (imac), uv-optimized gfp (gfpuv), enterokinase cleavage site for recovering hil-2 from purified fusion pro ... | 1999 | 10194406 |
| expression and purification of human interleukin-2 simplified as a fusion with green fluorescent protein in suspended sf-9 insect cells. | a fusion protein of human interleukin-2 (hil-2) and green fluorescent protein (gfp) was expressed in insect sf-9 cells infected with recombinant baculovirus derived from the autographa californica nuclear polyhedrosis virus (acnpv). this fusion protein was comprised of a histidine affinity ligand for simplified purification using immobilized metal affinity chromatography (imac), uv-optimized gfp (gfpuv) as a marker, an enterokinase cleavage site for recovery of hil-2 from the fusion, and the mod ... | 1999 | 10201111 |
| use of the baculovirus system to assemble polyomavirus capsid-like particles with different polyomavirus structural proteins: analysis of the recombinant assembled capsid-like particles. | the genes encoding the structural proteins (vp1, vp2 and vp3) of murine polyomavirus were cloned into the p2bac dual multiple cloning site vector, individually or jointly, and the corresponding proteins were expressed in spodoptera frugiperda (sf9) insect cells by cotransfecting sf9 cells with the constructed vector and the linear dna of autographa californica multiple nuclear polyhedrosis virus (acmnpv). recombinant capsid-like particles could be purified 5 days post-infection from sf9 cells in ... | 1999 | 10211971 |
| in vitro host range of autographa californica nucleopolyhedrovirus recombinants lacking functional p35, iap1 or iap2. | we have examined the host range in different insect cell lines of autographa californica nucleopolyhedrovirus (acmnpv) recombinants lacking p35, iap1 or iap2. these genes encode, or are predicted to encode, anti-apoptotic proteins. abrogation of p35 reduced the ability of acmnpv to replicate in permissive cell lines derived from spodoptera frugiperda insects by inducing apoptosis. in semi-permissive cell lines, such as lymantria dispar and spodoptera littoralis cells, we observed cytopathic effe ... | 1999 | 10211976 |
| autographa californica nuclear polyhedrosis virus dna polymerase: measurements of processivity and strand displacement. | the dna polymerase (dnapol) of autographa californica nuclear polyhedrosis virus was purified to homogeneity from recombinant baculovirus-infected cells. dnapol was active in polymerase assays on singly primed m13 template, and full-length replicative form ii product was synthesized at equimolar ratios of enzyme to template. the purified recombinant dnapol was shown to be processive by template challenge assay. furthermore, dnapol was able to incorporate hundreds of nucleotides on an oligo(dt)-p ... | 1999 | 10233952 |
| gene targeting in the silkworm by use of a baculovirus. | the bombyx mori fibroin light (l)-chain gene was cloned and the green fluorescent protein (gfp) gene inserted into exon 7. the chimeric l-chain-gfp gene was used to replace the polyhedrin gene of autographa californica nucleopolyhedrovirus (acnpv). this recombinant virus was used to target the l-chain-gfp gene to the l-chain region of the silkworm genome. female moths were infected with the recombinant virus and then mated with normal male moths. genomic dna from their progenies was screened for ... | 1999 | 10072379 |
| expression and structural characterization of a baculovirus ecdysteroid udp-glucosyltransferase. | the baculovirus enzyme ecdysteroid udp-glucosyltransferase (egt) disrupts the hormonal balance of the insect host by catalysing the conjugation of ecdysteroids, the moulting hormones, with the sugar moiety from udp-glucose or udp-galactose. in this study, egt has been overproduced using a recombinant autographa californica nucleopolyhedrovirus and an antiserum has been raised against the purified protein. this antiserum was used to visualize the kinetics of expression of egt by wild-type acmnpvl ... | 1999 | 10073711 |
| activation of baculovirus very late promoters by interaction with very late factor 1. | very late factor 1 (vlf-1) of autographa californica multicapsid nuclear polyhedrosis virus (acmnpv) activates the transcription of two genes, polyhedrin (polh) and p10, during the final, occlusion-specific phase of infection. using transient expression assays responsive to vlf-1, we identified linker scan mutations in the polh and p10 promoters which abolished or weakened the ability of the promoters to respond to stimulation by vlf-1. these mutations were located between the transcriptional an ... | 1999 | 10074194 |
| an immunological assay for determination of baculovirus titers in 48 hours. | th baculovirus expression system is a system of choice for expressing eukaryotic proteins. large amounts of biologically active material can be generated using this system by infecting insect cells with a baculovirus expressing the target protein. at several stages during the production of a baculovirus stock, it is necessary to titer the virus. current methods have long time lines and are either technically difficult or are limited to viruses expressing a reporter gene. the new assay described ... | 1999 | 10075805 |
| in vivo and in vitro analysis of baculovirus ie-2 mutants. | upon transient expression in cell culture, the ie-2 gene of autographa californica nuclear polyhedrosis virus (acmnpv) displays three functions: trans activation of viral promoters, direct or indirect stimulation of virus origin-specific dna replication, and arrest of the cell cycle. the ability of ie2 to trans stimulate dna replication and coupled late gene expression is observed in a cell line derived from spodoptera frugiperda but not in a cell line derived from trichoplusia ni. this finding ... | 1999 | 9971831 |
| requirement for gp64 to drive efficient budding of autographa californica multicapsid nucleopolyhedrovirus. | budded virions (bv) of autographa californica multicapsid nucleopolyhedrovirus (acmnpv) contain a major envelope glycoprotein (gp64) that is present on the plasma membrane of infected cells. gp64 is acquired by virions during budding through the plasma membrane, the final step in assembly of the budded virion at the cell surface. previous studies (s. a. monsma, a. g. p. oomens, and g. w. blissard (1996). j. virol. 70, 4607-4616) showed that insertional inactivation of the acmnpv gp64 gene result ... | 1999 | 9986796 |
| sequence and analysis of the genome of a baculovirus pathogenic for lymantria dispar. | the genome of the lymantria dispar multinucleocapsid nucleopolyhedrovirus (ldmnpv) was sequenced and analyzed. it is composed of 161,046 bases with a g + c content of 57.5% and contains 163 putative open reading frames (orfs) of >/=150 nucleotides. homologs were found to 95 of the 155 genes predicted for the autographa californica mnpv (acmnpv) genome. more than 9% of the ldmnpv genome was occupied by 16 repeated genes related to acmnpv orf2. readily identifiable homologs of several genes that h ... | 1999 | 9887315 |