Publications

TitleAbstractYear(sorted ascending)
Filter
PMID
Filter
rat monoclonal antibodies differentiating between the epstein-barr virus nuclear antigens 2a (ebna2a) and 2b (ebna2b).rat monoclonal antibodies were produced against the c-terminus of epstein-barr virus nuclear antigens 2a (ebna2a) and 2b (ebna2b) expressed as bacterial trpe fusion proteins. the initial screening was performed using a soluble bacterial extract containing the fusion proteins. positive hybridomas were confirmed by immunofluorescence on sf158 (spodoptera frugiperda) insect cells infected with recombinant baculovirus (autographa californica nuclear polyhedrosis virus) and expressing the complete eb ...199511831716
identification of a membrane fusion domain and an oligomerization domain in the baculovirus gp64 envelope fusion protein.the baculovirus gp64 envelope fusion protein (gp64 efp) is the major envelope glycoprotein of the budded virion and has been shown to mediate acid-triggered membrane fusion both in virions and when expressed alone in transfected cells. using site-directed mutagenesis and functional assays for oligomerization, transport, and membrane fusion, we localized two functional domains of gp64 efp. to identify a fusion domain in the gp64 efp of the orgyia pseudotsugata multiple nuclear polyhedrosis virus ...19957533858
influence of codon usage and translational initiation codon context in the acnpv-based expression system: computer analysis using homologous and heterologous genes.codon usage by all the known gene sequences from autographa californica nuclear polyhedrosis virus (acnpv) was compared with that of firefly luciferase (luc) and the beta subunit of human chorionic gonadotropin (beta hcg) expressed to contrasting levels in the baculovirus system. the highly expressed luc gene showed a codon usage similar to acnpv genes, as reflected by a very low d-squared statistic value (0.78) and a similar g/c usage (45%) at wobble positions. however, the underexpressed beta ...19957537424
isolation of p10 gene from bombyx mori nuclear polyhedrosis virus and study of its promoter activity in recombinant baculovirus vector system.a homologue of autographa californica npv (acnpv) p10 gene was identified and cloned from bombyx mori npv (bmnpv). bmnpv p10 gene encodes truncated protein of 70 amino acid residues that lacks carboxyl terminus comparing with the p10 protein encoded by acnpv. the putative tata box sequence and the ataag motif which is the consensus sequence of baculovirus very late promoter were conserved. a transfer vector, pbnt1, which includes the p10 promoter region of bmnpv for foreign gene expression was c ...19957547036
expression of human plasminogen activator inhibitor-2 in the baculovirus expression system.a cdna coding for human plasminogen activator inhibitor-2 (pai-2) was transferred into the genome of autographa californica nuclear polyhedrosis virus adjacent to the polyhedrin promoter. cells infected with recombinant virus synthesized a 43-kda pai-2 protein, the majority of which was secreted. the recombinant pai-2 was identical to native pai-2 purified from human placenta with respect to interaction with polyclonal antibody and inhibition of urokinase-type plasminogen activator.19957549934
production of functional human bone morphogenetic protein-2 using a baculovirus/sf-9 insect cell system.a clone of a human bone morphogenetic protein-2 (hbmp-2) cdna was obtained from a cdna library established from human dental pulp cells. after subcloning hbmp-2 cdna into autographa californica nuclear polyhedrosis virus, the recombinant baculovirus was transfected to sf-9 cells. immuno-reactive recombinant hbmp-2 (rhbmp-2) was detected by a polyclonal antibody against xenopus bmp-2 in the transfected insect cells but not in the culture media. three days after treatment with the lysate of the tr ...19957549938
biochemical analysis of the n-glycosylation pathway in baculovirus-infected lepidopteran insect cells.the baculovirus-insect cell system is used routinely for foreign glycoprotein production, but the precise nature of the n-glycosylation pathway in this system remains unclear. some studies indicate that these cells cannot process n-linked oligosaccharides to complex forms containing outer-chain galactose and sialic acid, while others indicate that they can. in this study, we used the major virion envelope glycoprotein of the baculovirus autographa california multicapsid nuclear polyhedrosis viru ...19957571420
replication of orgyia pseudotsugata baculovirus dna: lef-2 and ie-1 are essential and ie-2, p34, and op-iap are stimulatory genes.a transient dna replication assay was used to identify genes located within m.u. 90.5-7.0 of the orgyia pseudotsugata multinucleocapsid nuclear polyhedrosis virus (opmnpv) genome that influenced replication of a reporter plasmid containing an opmnpv origin of replication, when cotransfected into uninfected lymantria dispar cells. the viral transactivator ie-1 and a 2.4-kb subclone were found to be essential for replication. the 2.4-kb region was sequenced and open reading frames were identified. ...19957571435
identification and preliminary characterization of a chitinase gene in the autographa californica nuclear polyhedrosis virus genome.a functional chitinase gene (chia) has been identified in the genome of the autographa californica nuclear polyhedrosis virus (acmnpv). it is expressed in the late phase of virus replication in insect cells. high levels of both endo- and exochitinase activity were detected by 12 hr p.i. and remained stable throughout infection. an acmnpv chia protein-specific antibody was prepared using recombinant material prepared in bacteria. this was used to demonstrate that a product of approximately 58 kda ...19957571437
characterization, sequencing and phylogeny of the ecdysteroid udp-glucosyltransferase gene from two distinct nuclear polyhedrosis viruses isolated from choristoneura fumiferana.the ecdysteroid udp-glucosyltransferase (egt) gene isolated from a plaque-purified isolate of choristoneura fumiferana multinucleocapsid nuclear polyhedrosis virus (cfmnpv) was compared to its homologue from a defective mnpv virus (cfdef) present in wild type virus populations infecting the eastern spruce budworm, c. fumiferana. the egt genes were located in the same relative position within the virus genomes and their genomic location and arrangement were similar to that found in autographa cal ...19957595348
molecular structure and function of cd4 on murine egg plasma membrane.in the present study, the expression of the cd4 molecule on murine egg plasma membrane was confirmed by the indirect immunofluorescence (iif) method. the full-length cd4 cdna from murine eggs was synthesised by the reverse transcriptase-polymerase chain reaction (rt-pcr) method and its authenticity verified by southern blot hybridisation using an end-labelled internal oligonucleotide. the results of dna sequencing showed that the nucleotide sequence of the cdna of cd4 from murine egg mrna was id ...19957613876
identification, sequence, and transcriptional analysis of lef-3, a gene essential for orgyia pseudotsugata baculovirus dna replication.using a transient complementation assay for dna replication of an origin-containing reporter plasmid, a 2.4-kb region (m.u. 48.3-50.1) of the orgyia pseudotsugata multinucleocapsid nuclear polyhedrosis virus (opmnpv) genome was identified that contains a gene essential for opmnpv dna replication. this region was sequenced and open reading frames were identified. replication assays using subclones from this region identified a gene called late expression factor 3 (lef-3), as the essential replica ...19957618274
secretion of active human lecithin-cholesterol acyltransferase by insect cells infected with a recombinant baculovirus.difficulties in purifying the plasma enzyme lecithin-cholesterol acyltransferase (lcat) have hampered detailed studies of its (patho)physiological role in lipoprotein metabolism and of structure-function relationships. potentially, baculovirus-driven expression systems offer a powerful means to produce significant amounts of lcat. accordingly, full-length lcat cdna was cloned into pvl 1392, a high-level expression derivative of autographa californica nuclear polyhedrosis virus (acnpv), and the r ...19957619064
heterologous expression and enzymatic properties of a selenium-independent glutathione peroxidase from the parasitic nematode brugia pahangi.a full-length cdna from the parasitic nematode brugia pahangi encoding a secreted homolog of glutathione peroxidase in which the codon for the active site selenocysteine is substituted naturally by a cysteine codon has been expressed in spodoptera frugiperda (insect) cells via autographa californica nuclear polyhedrosis virus (baculovirus). the recombinant protein was glycosylated and secreted from the cells in tetrameric form. the purified protein showed glutathione peroxidase activity with a r ...19957629152
evaluation of protein-a linked monoclonal antibody latex agglutination test for diagnosis of nuclear polyhedrosis virus (bmnpv) of silkworm bombyx mori l.the symptomology of bmnpv infection in bombyx mori l depends on the stages of infection. discernible symptoms develop at later stages of infection, which leads to improper diagnosis and poor crop yield with sericulturists. in the present study development of direct and protein-a linked monoclonal antibody latex (palmal) agglutination test for the detection of bmnpv infection in silkworm is described. latex beads were precoated with protein-a and then sensitised with monoclonal antibody ma-231 (1 ...19957629276
construction and adhesive properties of a soluble madcam-1-fc chimera expressed in a baculovirus system: phylogenetic conservation of receptor-ligand interaction.madcam-1 is a high endothelial venule adhesion molecule composed of immunoglobulin and mucin-like domains which binds the leucocyte integrin lpam-1 (alpha 4 beta 7), and is largely responsible for the selective homing of lymphocytes to mucosal tissues. a novel soluble form of mouse madcam-1 which is normally membrane bound has been produced by joining the extracellular region of the receptor to the fc domain of human igg1. the madcam-1-fc cdna was inserted into the genome of autographa californi ...19957631157
baculovirus-driven expression and purification of glycine receptor alpha 1 homo-oligomers.the glycine receptor is a ligand-gated anion channel protein of postsynaptic membranes. we expressed a homo-oligomeric receptor composed of human alpha 1 subunits in spodoptera frugiperda cells by infection with a recombinant autographa californica nuclear polyhedrosis virus. a substantial fraction of the recombinant receptor was incorporated as a functional channel protein into the cell's plasma membrane at expression levels 4- to 30-fold higher than in other eukaryotic heterologous expression ...19957635206
expression of pe38 and ie2, viral members of the c3hc4 finger family, during baculovirus infection: pe38 and ie2 localize to distinct nuclear regions.the pe38 gene of autographa californica nuclear polyhedrosis virus represents one of the major early transcripts after viral infection. the function of the pe38 protein, which contains a c3hc4 zinc finger motif, is still not understood. we have raised polyclonal antiserum against the pe38 protein, pe38, produced in bacteria to investigate pe38 expression in the course of infection. a approximately 38-kda polypeptide is first detectable at 2 h postinfection and decreases rapidly after 24 h. durin ...19957636971
transcriptional enhancer activity of hr5 requires dual-palindrome half sites that mediate binding of a dimeric form of the baculovirus transregulator ie1.the hr5 enhancer element stimulates early viral transcription and may function as an origin of dna replication for autographa californica nuclear polyhedrosis virus (acmnpv). the smallest functional unit of hr5 is a 28-bp repeat consisting of an imperfect palindrome (28-mer). to identify essential sequences and examine the molecular basis of hr5 activity, the effects of site-directed mutations on transcriptional enhancement by the 28-mer and binding of the acmnpv transregulator ie1 were investig ...19957636981
baculovirus-mediated production of the human growth hormone in larvae of the silkworm, bombyx mori.to express the cdna encoding human growth hormone (hgh) in larvae of bombyx mori, b. mori nuclear polyhedrosis virus (bmnpv) was employed as an expression vector. for the construction of the recombinant virus, the hgh cdna was inserted into the downstream of the strong polyhedrin promoter to achieve a high level expression. immunoblot analysis revealed that the virus-mediated hgh was synthesized in the larvae and secreted into the hemolymph. the yield of the recombinant hgh synthesized in the la ...19957646491
differential requirements for baculovirus late expression factor genes in two cell lines.a plasmid library of 18 late expression factor (lef) genes (lef library) from the baculovirus autographa californica nuclear polyhedrosis virus (acmnpv) supports transient expression from a late viral promoter in the sf-21 cell line, derived from spodoptera frugiperda. we found, however, that this lef library was unable to support expression from the same promoter in the tn-368 cell line, derived from trichoplusia ni, which is also permissive for acmnpv replication. to identify the additional fa ...19957666527
baculovirus mediated high-level expression of luciferase in silkworm cells and larvae.bombyx mori nuclear polyhedrosis virus (bmnpv)-based baculovirus expression system exploits silkworm larvae as an economical alternative to large-scale cell cultures for production of biomolecules. to generate recombinant bmnpv at high frequencies, we have achieved high efficiency transfection of b. mori cells, bmn, through lipofection. optimal conditions for lipofection were standardized by quantification of the transient expression level of firefly luciferase (luc) reporter gene under control ...19957669303
use of juvenile hormone esterase as a novel reporter enzyme in the baculovirus expression system.juvenile hormone esterase (jhe) has a number of characteristics favorable for use as a reporter enzyme. it is extremely stable under a variety of adverse conditions including in organic solvent. jhe is easily detected by a rapid and sensitive colorimetric assay, and detection is facilitated by export of the enzyme from the cell. its use is illustrated in the baculovirus expression system by promoter studies and evaluation of culture conditions necessary for optimal production of recombinant prot ...19957730431
characterization of v-cath, a cathepsin l-like proteinase expressed by the baculovirus autographa californica multiple nuclear polyhedrosis virus.autographa californica multiple nuclear polyhedrosis virus (acmnpv) contains a 966 bp orf that encodes a papain type cysteine proteinase with cathepsin l-like characteristics. using western blot analysis of infected cell extracts we showed that v-cath proteinase has 35.5 kda and 32 kda precursor forms which are processed to a 27.5 kda mature form in a manner characteristic of papain and cathepsin l. v-cath proteinase activity was greatest under acidic conditions (ph 5.0) and was reduced in the p ...19957730794
overexpression of bombyx mori prothoracicotropic hormone using baculovirus vectors.recombinant baculoviruses were constructed that express the cdna encoding the prothoracicotropic hormone (ptth) of bombyx mori. this hormone stimulates the production of ecdysteroids by the insect's prothoracic glands. two groups of viruses were constructed, expressing either the entire cdna encoding prepro-ptth, or a synthetic chimeric gene encoding a signal peptide fused to the mature ptth subunit. in both cases, the genes were expressed in wild-type autographa californica nuclear polyhedrosis ...19957742834
the cagt motif functions as an initiator element during early transcription of the baculovirus transregulator ie-1.the highly conserved tetranucleotide cagt is located at the rna start site of the transregulator gene ie-1 of autographa californica nuclear polyhedrosis virus (acmnpv). the presence of this motif within numerous baculovirus early promoters and its similarity to transcriptional initiators suggested a fundamental role in viral transcription regulation. to determine the function of the cagt motif, site-specific mutations were introduced within the ie-1 promoter fused to a reporter gene within acmn ...19957745705
the lef-3 gene of autographa californica nuclear polyhedrosis virus encodes a single-stranded dna-binding protein.the autographa californica nuclear polyhedrosis virus (acnpv) replicates in the nuclei of infected cells and encodes several proteins required for viral dna replication. as a first step in the functional characterization of viral replication proteins, we purified a single-stranded dna-binding protein (ssb) from acnpv-infected insect cells. nuclear extracts were chromatographed on single-stranded dna agarose columns. an abundant protein with an apparent molecular weight of 43,000 was eluted from ...19957745748
the autographa californica nuclear polyhedrosis virus homologous region 1a: identical sequences are essential for dna replication activity and transcriptional enhancer function.hr1a consists of two 30-bp imperfect palindrome sequences separated by 58 bp and each palindrome contains a naturally occurring ecori site at its core. plasmid subclones of the hr1a-containing acmnpv hindiii-n fragment were examined for their ability to replicate in virus-infected (spodoptera frugiperda) sf9 cells, and to stimulate transcription when linked in cis with a 39k gene promoter-beta-glucuronidase fusion and cotransfected into cells along with a plasmid (ple-1) containing the gene enco ...19957747446
a temperature-sensitive ie1 protein of autographa californica nuclear polyhedrosis virus has altered transactivation and dna binding activities.the temperature-sensitive mutant tsb821 of autographa californica nuclear polyhedrosis virus has two nucleotide transitions in the transregulatory ie1 gene (ribiero et al. (1994) j. virol. 68, 1075-1084). these mutations result in two conservative amino acid substitutions in the encoded protein. to determine whether the mutations affect the ability of ie1 to interact with viral enhancer elements, electrophoretic mobility shift assays were performed using extracts prepared from infected cells and ...19957747488
expression of the ie1 transactivator of autographa californica nuclear polyhedrosis virus during viral infection.the immediate-early ie1 protein of autographa californica nuclear polyhedrosis virus (acmnpv) is an important regulator of viral gene transcription. to provide a tool for further analysis of the expression and function of ie1, a polyclonal antiserum was raised against ie1 expressed in bacteria. immunoblot analysis of infected cell lysates was used to monitor the accumulation of ie1 throughout the viral life cycle. when extracts were prepared in the presence of phosphatase inhibitors, only one pr ...19957747489
synthesis and processing of the rubella virus p110 polyprotein precursor in baculovirus-infected spodoptera frugiperda cells.in order to study the processing of rubella virus (rv) structural proteins (capsid protein, of 33 kda; e2 of 42-47 kda; and e1 of 58 kda) in spodoptera frugiperda (fall armyworm) cells, a 24s cdna encoding the polyprotein precursor, p110, was inserted under the transcriptional regulation of the polyhedrin gene promoter of the autographa californica nuclear polyhedrosis virus (acnpv) and expressed during viral infection. by immunoblot analysis using antibodies directed against whole rv and the in ...19957754676
p10 genes of bombyx mori nuclear polyhedrosis virus and autographa californica multiple nuclear polyhedrosis virus.ecor i-p fragment has been cloned from autographa californica multiple nuclear polyhedrosis virus (acmnpv) genomic dna and used as a probe. 0.5-kb and 1.1-kb fragments including p10 gene from bombyx mori nuclear polyhedrosis virus (bmnpv) have been hybridized. the p10 orf was located in the ecor i-r fragment. initiation codon atg of p10 from bmnpv has been mutated by pcr, and the atg region became a bgl ii site. a novel transfer vector pbmacpv-1 has been constructed using both the p10 5'-flankin ...19957695816
efficient gene transfer into human hepatocytes by baculovirus vectors.viral vectors are the most efficient tools for gene delivery, and the search for tissue-specific infecting viruses is important for the development of in vivo gene therapy strategies. the baculovirus autographa californica nuclear polyhedrosis virus is widely used as a vector for expression of foreign genes in insect cells, and its host specificity is supposed to be restricted to arthropods. here we demonstrate that recombinant a. californica nuclear polyhedrosis virus is efficiently taken up by ...19957479733
beet western yellows luteovirus capsid proteins produced by recombinant baculoviruses assemble into virion-like particles in cells and larvae of bombyx mori.we subcloned various constructs of the beet western yellows luteovirus (bwyv) 3' proximal genes into bombyx mori nuclear polyhedrosis virus (bmnpv) transfer vectors and constructed recombinant bmnpvs. the recombinant bmnpvs were used to infect bmn cells and b. mori larvae. protein expression was analyzed by sds-page and immunoblot analysis, and the bwyv-encoded capsid protein (cp), p19, readthrough protein (rt), and a modified capsid-readthrough protein (cp-rt*) were detected in bmn cells infect ...19957483264
functional expression of an alpha anti-insect scorpion neurotoxin in insect cells and lepidopterous larvae.the leiurus quinquestriatus hebraeus alpha anti-insect toxin (lqh alpha it) cdna was engineered into the autographa californica nuclear polyhedrosis virus (acnpv) genome. insect cells infected with the recombinant virus secreted a functional lqh alpha it polypeptide. spodoptera littoralis and heliothis armigera larvae injected with recombinant budded virus, showed typical intoxication symptoms. this recombinant virus showed enhanced insecticidal potency against h. armigera larvae compared with w ...19957498537
transcriptional regulation of cell line-dependent, baculovirus-mediated expression of foreign genes.cell line-dependent expression of foreign genes in the baculovirus system was investigated using a recombinant vac beta hcg-luc virus carrying two reporter genes--beta subunit of human chorionic gonadotropin (beta hcg) and luciferase (luc)--placed under the transcriptional control of the autographa californica nuclear polyhedrosis virus (acnpv) polyhedrin gene promoters. five different lepidopteran cell lines derived from spodoptera frugiperda (sf21 and sf9), bombyx mori (bmn and bm5), and trich ...19957530452
high level synthesis and secretion of human urokinase using a late gene promoter of the autographa californica nuclear polyhedrosis virus.a cdna encoding human urokinase was inserted into the autographa californica nuclear polyhedrosis virus (acnpv) genome at the polyhedrin gene locus under control of a duplicated copy of the late, basic protein gene promoter. the insect-derived urokinase was produced predominantly in the form of single-chain, pro-urokinase, with a molecular mass of 50 kda, and demonstrated fibrinolytic activity. synthesis and secretion of urokinase was first detected at 6 hours post-infection and continued steadi ...19957766008
deletion of the baculovirus ecdysteroid udp-glucosyltransferase gene induces early degeneration of malpighian tubules in infected insects.deletion of the ecdysteroid udp-glucosyltransferase gene (egt) from the autographa californica nuclear polyhedrosis virus (acnpv) genome increases the speed of killing of this virus (d. r. o'reilly and l. k. miller, bio/technology 9:1086-1089, 1991). second-instar spodoptera exigua larvae are killed more rapidly by the egt deletion mutant of acnpv than by wild-type acnpv. unlike wild-type acnpv-infected larvae, larvae infected with an egt deletion mutant molt and resume feeding as mock-infected ...19957769717
properties of a baculovirus mutant defective in the protein phosphatase gene.autographa california nuclear polyhedrosis virus (acmnpv) contains a gene, ptp, encoding a protein tyrosine/serine phosphatase, bv-ptp. to investigate the biological function of ptp in the baculoviral replication cycle, we constructed a recombinant baculovirus, vptpdel, in which the catalytically active site of bv-ptp was deleted. although the vptpdel mutant was viable in cell culture, it was partially defective in occluded virus production in sf-21 but not tn-368 cell lines. sf-21 cells infecte ...19957769718
the baculovirus transactivator ie1 binds to viral enhancer elements in the absence of insect cell factors.the transregulatory ie1 protein of autographa californica nuclear polyhedrosis virus binds to the viral enhancer element hr5. to test whether ie1 binds independently of host cell factors, ie1 was translated in rabbit reticulocyte extracts and tested for dna binding activity by an electrophoretic mobility shift assay. complexes with the hr5 probe were detected with translation reaction mixtures primed with ie1 rna but not with control translation reaction mixtures. however, the dna-protein comple ...19957769721
n-glycosylated polypeptides synthesized in the early phase of acnpv infected insect cells.infected cell specific polypeptides (icsp) in autographa californica nuclear polyhedrosis virus (acnpv) infected spodoptera frugiperda cell line, sf9, were radiolabelled with [14c] mannose and the labelled proteins were monitored at various time intervals. a major glycoprotein of molecular size of 64 kda appears as early as 3h post infection (p.i.) which peaks at 12h p.i., but in presence of tunicamycin this protein is not labelled. two other glycoproteins of molecular size 67 kda and 63 kda app ...19957773189
developmental resistance in fourth instar trichoplusia ni orally inoculated with autographa californica m nuclear polyhedrosis virus.larvae of lepidopteran insects commonly become increasingly resistant to baculovirus infections as they age. the mechanism responsible for this development resistance is not known, but the phenomenon does not occur if the viral inoculum is administered intrahemocoelically instead of orally, which is the natural route of infection. this observation indicates that the factors mediating developmental resistance are operative during infection of the primary target tissue, the larval midgut, and not ...19957778273
identification and analysis of a putative origin of dna replication in the choristoneura fumiferana multinucleocapsid nuclear polyhedrosis virus genome.a recombinant plasmid containing the choristoneura fumiferana multinucleocapsid nuclear polyhedrosis virus (cfmnpv) hindiii r fragment (m.u. 2.2-3.9) was shown to undergo cfmnpv infection-dependent dna replication in cf-124t cells. replication of this dna sequence was detectable by 24 hr p.i. and did not appear to have resulted as a consequence of recombination with the virus genome. replication was inhibited by mimosine, an inhibitor of eukaryotic dna replication. these data suggest that hindii ...19957778276
the wild-type autographa californica nuclear polyhedrosis virus induces apoptosis of spodoptera littoralis cells.spodoptera littoralis cells infected with the autographa californica multiple nuclear polyhedrosis virus (acmnpv) yielded significantly lower budded virus titers than spodoptera frugiperda-infected cells and produced very low levels of polyhedrin. relative to acmnpv-infected s. frugiperda sf9 cells viral dna replication was severely reduced in spodoptera littoralis sl2 cells. microscopic examination of sl2-infected cells revealed progressive cell blebbing starting at 6-8 hr postinfection and cul ...19957778284
identification, localization, transcription, and sequence analysis of the choristoneura fumiferana nuclear polyhedrosis virus dna polymerase gene.the location of the choristoneura fumiferana baculovirus dna polymerase gene was determined by hybridization analysis using a probe prepared from the previously identified polymerase gene from the autographa californica multiple nuclear polyhedrosis virus. dna sequence analysis revealed that the choristoneura fumiferana baculovirus dna polymerase gene consists of 2970 base pairs encoding 990 amino acids (114.2 kda). transcriptional analysis demonstrated that overlapping transcripts of 3.2 and 4. ...19957778286
comparative pathogenesis of autographa californica m nuclear polyhedrosis virus in larvae of trichoplusia ni and heliothis virescens.we compared early viral pathogenesis and dose-mortality relationships for larvae of two highly susceptible hosts, trichoplusia ni and heliothis virescens, using a construct of acmnpv containing the lacz reporter gene. larvae were inoculated either as newly molted fourth instars (4(0)) or 15 hr after the molt (4(15)). in 4(0)-inoculated larvae, first lacz expression was detected in the midgut epithelium of t. ni at 4 hr postinoculation (hpi) compared to 18 hpi in h. virescens, and systemic infect ...19957778288
the baculovirus gp64 envelope fusion protein: synthesis, oligomerization, and processing.the baculovirus gp64 envelope fusion protein (gp64 efp) is a class i integral membrane protein that enters the secretory pathway and is oligomerized and extensively processed during transport to the plasma membrane. the kinetics of gp64 efp biosynthesis, oligomerization, and processing in orgyia pseudotsugata multicapsid nuclear polyhedrosis virus (opmnpv)-infected lymantria dispar cells were examined by pulse label, pulse-chase, and immunoprecipitation experiments. relative rates of gp64 efp sy ...19957778291
nucleotide sequence and transcriptional analysis of the gp41 gene of spodoptera frugiperda nuclear polyhedrosis virus.the spodoptera frugiperda multiple nucleocapsid nuclear polyhedrosis virus (sfmnpv) gp41 structural protein gene was located in the 1.9 kbp ecori-s fragment and sequenced. an open reading frame (orf) of 999 nucleotides was detected that encoded a protein of 332 amino acids. the gp41 gene transcript was detected after 12 h post-infection (p.i.) and remained detectable at 48 h p.i. two major mrnas, about 1.6 and 2.8 kb in length, were determined by northern blot analysis. primer extension analysis ...19957782772
mutations in the autographa californica multinucleocapsid nuclear polyhedrosis virus 25 kda protein gene result in reduced virion occlusion, altered intranuclear envelopment and enhanced virus production.serial passage of nuclear polyhedrosis viruses (npvs) through cultured cell lines results in the appearance of mutants with a complex phenotype referred to as the 'few polyhedra' (fp) phenotype. the altered plaque morphology and reduced occlusion production associated with the fp phenotype have been observed in autographa californica multinucleocapsid nuclear polyhedrosis virus (acmnpv) bearing mutations in the gene encoding the 25 kda protein (25k gene). in this study, we sequenced the 25k gene ...19957782773
expression of human bone morphogenetic protein 2 in bombyx mori larvae and its purification.recombinant bm-bacpak-bmp2 virus which contains the human bone morphogenetic protein 2 (bmp2) cdna gene under the control of polyhedrin gene promoter was generated by cotransfection into bm-n cells with the transfer plasmid pbacpak-bmp2 and the modified bombyx mori nuclear polyhedrosis virus bm-bacpak genomic dna. on the fifth day after infection with the recombinant virus, bombyx mori larvae produced about 10 &mgr;g rbmp2 in 1 ml hemolymph. the expressed rbmp2 was processed into 16 kd c-termina ...199612232629
sequencing and structural analysis of a novel early-late gene of the leucania separata nuclear polyhedrosis virus. 199612237712
molecular structure of a proteolytic fragment of tlp20.myosin light-chain kinase is responsible for the phosphorylation of myosin in smooth muscle cells. in some tissue types, the c-terminal portion of this large enzyme is expressed as an independent protein and has been given the name telokin. recently, an antibody directed against telokin was found to interact with a protein derived from the baculovirus autographa californica nuclear polyhedrosis virus. this protein was biochemically characterized and given the name tlp20 for telokin-like protein ...199615299576
activity and persistence of steinernema carpocapsae and spodoptera exigua nuclear polyhedrosis virus against s. exigua larvae on soybean.experiments were conducted to assess the effects of the entomopathogenic nematode steinernema carpocapsae and spodoptera exigua multinucleocapsid nuclear polyhedrosis virus (semnpv), alone and in combinations, on mortality of the beet armyworm, s. exigua, larvae on soybean. in 1991 tests, field-grown soybean plants were treated with s. carpocapsae at 0.3 and 0.6 nematodes/cm(2) of leaflet, semnpv at 20 and 40 polyhedral inclusion bodies (pib)/cm(2), and all possible combinations. treated leaflet ...199619277347
in vivo production, stabilization, and infectivity of baculovirus preoccluded virions.wild-type and polyhedrin-negative isolates of autographa californica nuclear polyhedrosis virus were replicated in fifth-instar trichoplusia ni larvae. insect tissues infected with wild-type virus contained two types of virions that are highly infectious when ingested, those occluded in polyhedra and preoccluded virions. tissue infected with the polyhedrin-negative virus contained only preoccluded virions. the relative potencies of the two types of infected tissue were determined by dose-mortali ...199616535202
abortive infection of the baculovirus autographa californica nuclear polyhedrosis virus in sf-9 cells after mutation of the putative dna helicase gene.homologous recombination between the autographa californica nuclear polyhedrosis virus (acnpv) genome and a 0.6-kbp-long dna fragment derived from the putative dna helicase gene of bombyx mori nuclear polyhedrosis virus generates eh2-acnpv, an expanded-host-range acnpv mutant (s. maeda, s.g. kamita, and a. kondo, j. virol. 67:6234-6238, 1993). after inoculation at a high multiplicity of infection (moi), eh2-acnpv replicates efficiently in both the sf-9 (acnpv-permissive) and bmn (non-acnpv-permi ...19968709251
nucleotide sequence, insertional mutagenesis, and transcriptional mapping of a conserved region of the baculovirus autographa californica nuclear polyhedrosis virus (map unit 64.8-66.9).the nucleotide sequence of a 2773-bp region of the autographa californica nuclear polyhedrosis virus hr3 variant (acmnpv) (map unit 64.4-68.2) has been determined. this region lies between the previously analyzed p6.9 and p80 genes. three open reading frames were contained within this region, potentially coding for proteins of 45, 40, and 12 kda. the 5' ends of transcripts capable of coding for the 45- and 12-kda proteins map to consensus baculovirus late transcription start sites (ataag), while ...19968989865
[expression of cdna of human chorionic gonadotropin beta-subunit (beta-hcg) cdna in insect cells and effect of expressed product on mouse lymphocytes in vitro].expression vector pvl 1393-hcg beta containing beta-hcg cdna has been constructed using an unfused protein nuclear polyhedrosis virus (acnpv) expression vector. the insect cells (sf 9) were cotransfected by the expression vector and nuclear polyhedrosis virus genomic dna, and recombinant virus acnpv-hcg beta was screened out, beta-hcg cdna was expressed in insect cells infected by recombinant virus and recombinant beta-hcg (r beta-hcg) was secreted into medium. the purity of r beta-hcg, purified ...19969208644
secretion of green fluorescent protein from recombinant baculovirus-infected insect cells.trichoplusia ni (high five) and spodoptera frugiperda (sf21) cells were engineered for expression of epitope (flag)-tagged signal peptide-green fluorescent protein (gfp) fusions to examine the suitability of gfp as a secretory marker. the recombinant baculovirus-infected cells became fluorescent, and the high five cells but not sf21 cells secreted gfp in the culture medium as detected by the presence in the culture supernatant of a flag-immunoreactive 30-kda species and the characteristic 510-nm ...19968831686
bifunctionality of the acmnpv homologous region sequence (hr1): enhancer and ori functions have different sequence requirements.the autographa californica multinucleocapsid nuclear polyhedrosis virus (acmnpv) homologous region sequence hr1 is a putative origin of replication (ori) sequence and can also function as a transcriptional enhancer for delayed-early genes. we demonstrate that this 750-bp sequence, carrying five 28-bp core palin-dromes, enhances expression from the very late polyhedrin promoter up to 11-fold in a classical enhancer fashion in transient expression assays. enhancement is at the level of transcripti ...19968836032
in vitro characterization of a trichoplusia ni single nucleocapsid nuclear polyhedrosis virus.a trichoplusia ni single nucleocapsid nuclear polyhedrosis virus (tnsnpv) isolate was cloned and its replication studied in the bti-tn-5b1-4 insect cell line. the bti-tn-5b1-4 cells were highly susceptible to tnsnpv infection, with 99% of the cells containing viable polyhedra by 30 h post-inoculation. viral dna synthesis was detected by 9 h post-infection (p.i.). infectious budded virus (bv) was first detected at 13 h p.i. and reached an average maximum titre of 3.875 x 10(6) p.f.u./ml 27 h p.i. ...19968811031
continuous cell line from pupal ovary of perina nuda (lepidoptera: lymantriidae) that is permissive to nuclear polyhedrosis virus from p. nudaa continuous cell line, designated ntu-pn-hh, was established from the pupal ovary of perina nuda fabricius (lepidoptera: lymantriidae). the cells have been through more than 300 passages during 4 years in tnm-fh medium supplemented with 10% fbs, at a constant temperature of 28°c. the cell line consists of four major morphologic types: polymorphic cells, spindle-shaped cells, round cells, and squamous cells. the characterization of this cell line showed that ntu-pn-hh is a newly established ...19968812599
expression and analysis of a bacterial poly(hydroxyalkanoate) synthase in insect cells using a baculovirus system.an improved method for expression of poly-beta-hydroxyalkanoate (pha) synthase from alcaligenes eutrophus has been developed using the baculovirus autographa californica nuclear polyhedrosis virus (acmnpv) in bti-tn-5b1-4 trichoplusia ni cells which results in high level production of active pha synthase. confirmation of expression of authentic pha synthase was obtained by western analysis which also revealed the presence of several apparent proteolytic cleavage products. n-terminal sequence dat ...19968812862
the genes encoding the p39 and cg30 proteins of bombyx mori nuclear polyhedrosis virus.we have cloned and analysed the transcriptional properties of two closely linked genes, p39 and cg30, of bombyx mori nuclear polyhedrosis virus (bmnpv). these genes encode a structural polypeptide and a putative transcriptional regulator of the virus, respectively. the cg30 gene is transcribed prior to and after dna replication from a site located within the orf of the adjacent p39 gene. its transcription product, a 1.3 kb mrna, is polyadenylated at a site containing consensus eukaryotic polyade ...19969000108
genetic variability of four natural isolates of the stilpnotia salicis multiple-enveloped nuclear polyhedrosis virus.viral genome dna from four different multiple-enveloped nuclear polyhedrosis virus isolates, obtained from naturally infected larvae of satin moth (stilpnotia salicis), a pest of poplar tree (populus) was analysed. larvae were collected over a period of 11 years, from 1978 to 1989. the genomic dna restriction patterns pointed to heterogeneity of these wild-type viruses. the differences observed in isolates of several years revealed limited restriction fragment length polymorphism and showed that ...19969104499
expression of human growth hormone in silkworm larvae through recombinant bombyx mori nuclear polyhedrosis virus.we have generated a recombinant bombyx mori nuclear polyhedrosis virus, vbmhgh, harboring the full length human growth hormone gene (2.4-kb genomic dna, with four introns and the signal peptide sequences) under the control of the polyhedrin promoter. bmn cells in culture infected with the recombinant virus showed the presence of rna corresponding to the authentic growth hormone mrna as well as its incompletely processed precusor. electrophoretic analysis and immunoprecipitation of proteins of re ...19968860651
temperature-sensitive mutations in the protein kinase-1 (pk-1) gene of the autographa californica nuclear polyhedrosis virus that block very late gene expression.we have developed a genetic screen for temperature-sensitive mutations in the very late transcription apparatus of the autographa californica nuclear polyhedrosis virus. this method starts with the bacpaks virus, which has the escherichia coli lacz gene under the control of the very late polyhedrin promoter. the desired mutants are temperature-sensitive for beta-galactosidase production and can be complemented by wild-type virus, which lacks the lacz gene. two mutants created by nitrosoguanidine ...19968862393
structure-function analysis of the autographa californica multinucleocapsid nuclear polyhedrosis virus homologous region palindromes.homologous regions (hrs), which are present at eight dispersed locations on the autographa californica multinucleocapsid nuclear polyhedrosis virus genome, are composed of repeated imperfect palindromes within directly repeated sequences. hrs act as transcriptional enhancers of rna polymerase ii-mediated transcription and as origins of dna replication when incorporated into plasmids and tested in transient replication assays. to characterize the physical structure of these elements and to determ ...19968862418
a 38-kda host factor interacts with functionally important motifs within the autographa californica multinucleocapsid nuclear polyhedrosis virus homologous region (hr1) dna sequence.we recently demonstrated that the autographa californica multinucleocapsid nuclear polyhedrosis virus homologous region (hr1) enhances transcription from the viral polyhedrin promoter and also functions as a putative origin of replication (ori). hr1, carrying five 28-base pair core palindrome units, has also been mapped with respect to its enhancer and ori functions (habib, s., pandey, s., chatterji, u., burma, s., ahmad, r., jain, a., and hasnain, s. e. (1996) dna cell biol. 15, 737-747). a 38- ...19968910443
a baculovirus vector derived from immediately early gene promoter of autographa californica nuclear polyhedrosis virus.a transfer vector was constructed in which the neomycin resistance (neo) gene was under the control of a copy of autographa californica nuclear polyhedrosis virus (acmnpv) ie1 gene promoter at the p10 locus. after cotransfection of spodoptera frugiperda (sf9) cells with the transfer vector and infectious acmnpv dna, the recombinant virus-containing neo gene was selected by serial passage of the mixed progenies from cotransfection. this was done at low moi in the presence of g418 in growth medium ...19968910650
expression and loading of recombinant heavy and light chain homopolymers of rat liver ferritin.the full-length genes for the heavy (h) and light (l) chains of ferritin isolated from a rat liver cdna library were amplified using polymerase chain reaction. each was inserted at the unique bglii site downstream of the p10 promoter of the baculovirus transfer vector pacuw21. the genes were transferred separately to infectious autographa californica nuclear polyhedrosis virus (acnpv) expression vectors after in vivo homologous recombination. ferritin homopolymers of either h or l chain were exp ...19968914851
overexpression of actin in acmnpv-infected cells interferes with polyhedrin synthesis and polyhedra formation.autographa californica m nuclear polyhedrosis virus (acmnpv) terminates host protein synthesis during the late stage of infection, at approximately 14 hr postinfection (hpi). if infection takes place in the presence of cytochalasin d (cd), however, host actin synthesis is transiently stimulated and continues to be synthesized until approximately 30 hpi, and the hyperexpression of polyhedrin is delayed from about 20 hpi until about 36 hpi (s. n. talhouk and l. e. volkman, virology 182, 626-634, 1 ...19968918923
cloning and sequencing of the gene for an enhancing factor from pseudaletia separata entomopoxvirus.an enhancing factor (ef) has been shown to increase the susceptibility of the armyworm, pseudaletia separata, to this nuclear polyhedrosis virus. using amino acid microsequencing data obtained from the purified ef protein, we cloned and sequenced the gene for ef. the deduced amino acid sequence exhibited a significant similarity to major spindle proteins, fusolins, whose function is unknown, from heliothis armigera, choristoneura biennis and melolontha melolontha entomopoxviruses.19968921880
gene transfer into hepatocytes and human liver tissue by baculovirus vectors.gene therapy of liver diseases requires the development of efficient vectors for gene transfer in vivo. retroviral and adenoviral vectors have been shown to deliver genes efficiently into hepatocytes in vitro and in vivo. however, these vectors do not allow for exclusive infection of the liver which would be highly advantageous for in vivo gene therapy strategies. we have recently demonstrated that genetically modified baculoviruses (autographa californica nuclear polyhedrosis virus) efficiently ...19968930653
the role of the acmnpv 25k gene, "fp25," in baculovirus polh and p10 expression.a previous study showed that an autographa californica multicapsid nuclear polyhedrosis virus (acmnpv) 25k mutant produced less polyhedrin protein than wild-type (jarvis et al., j. virol. 66, 6903-6911, 1992). in this study, the role of the 25k gene product (acmnpv orf 61) in baculovirus gene expression was further investigated. five different viral 25k mutants expressed lower levels of polyhedrin protein and less cat activity under the control of the polh promoter compared to wild-type. polh rn ...19968941320
inhibition of interleukin 1 beta-converting enzyme-mediated apoptosis of mammalian cells by baculovirus iap.apoptosis can be a potent weapon against viral infection and consequently has selected for viruses carrying antiapoptosis genes. two baculovirus proteins, iap and p35, can prevent insect cells from dying in response to infection. p35, which interferes with members of the ced-3 family of cysteine proteases, can also function in mammalian cells. we investigated the ability of iap from orgyia pseudotsugata nuclear polyhedrosis virus to prevent death of mammalian cells. iap was transiently expressed ...19968943013
effects of viral inhibitors of apoptosis in models of mammalian cell death.apoptotic cell death is used as a defence against infection by viruses. to counter this protective mechanism, some viruses carry genes whose products can inhibit progression of the apoptotic process in the host cell. as it is clear that the core cell death mechanisms have been conserved through evolution, viral genes from various sources can be used to unravel these mechanisms in mammalian cells. we have produced transgenic mice that express the cowpox gene crma in their t cell compartment, and ...19968950470
expression of biologically active human granulocyte-macrophage colony-stimulating factor in the silkworm (bombyx mori).using virus derived from bombyx mori (silkworm) nuclear polyhedrosis virus (bmnpv), we constructed an infectious recombinant virus carrying the human granulocyte-macrophage colony-stimulating factor (hgm-csf) cdna placed downstream from the polyhedrin promoter. the bmn cells infected with the recombinant baculovirus bmnpv-gm-csf expressed recombinant (r)hgm-csf up to 3.9 x 10(5) colony-formation units/ml in the medium. the maximum activity of rhgm-csf was 4.3 x 10(6) colony-formation units/ml in ...19968969454
a variable region of anticarsia gemmatalis nuclear polyhedrosis virus contains tandemly repeated dna sequences.a variable region (psti-t fragment) of two genotypic variants of the anticarsia gemmatalis multiple nucleocapsid nuclear polyhedrosis virus (agmnpv) was sequenced and compared. this region, which is known to have deletions and duplications in agmnpv variants, was shown to be formed of units of a 127 bp tandemly repeated sequence containing two 30 bp imperfect palindromes. southern blot experiments showed that the psti-t fragment contained one of the four homologous regions mapped interspersed in ...19968738171
[heterologous protein expression in the baculovirus vector system: the nuclear polyhedrosis virus of malacosoma neustria in antheraea pernyi cells].the results of the creation of new expressive system on malacosoma neustria nuclear polyhedrosis virus and cell line originated from antheraea pernyi tissues was demonstrated. the expression of bacterial beta-galactosidase-coding gene was analyzed in this system.19968743017
construction of an insecticidal baculovirus expressing insect-specific neurotoxin aait.considering the factors which affect gene transcription, translation and the stability of mrna, without changing the amino acid composition of the encoded polypeptide, aait gene encoding insect-specific neurotoxin was designed and synthesized according to bias in codon choice, overall g+c content and g+c content of bases at the third position in codons of polyhedrin genes of baculovirus and of plant genes as well. aait gene was fused behind a synthetic gp67 signal sequence and then recombined in ...19968760466
species-specific effects of the hcf-1 gene on baculovirus virulence.the host cell-specific factor 1 gene (hcf-1) of the baculovirus autographa californica nuclear polyhedrosis virus (acmnpv) is required for the efficient replication and/or stability of reporter plasmids carrying an acmnpv-derived origin of dna replication in a cell-specific manner; hcf-1 is required for reporter plasmid replication or stability in tn-368 cells, a cell line derived from the cabbage looper trichoplusia ni, but not in iplb-sf-21 (sf-21) cells, a cell line derived from the fall army ...19968764020
preparation and characterization of two human carcinoembryonic antigen family proteins of neutrophils, cd66b and c, in silkworm larvae.as a step to investigate the cell adhesion mechanism and physiological roles of two cd66 antigens in human neutrophils, carcinoembryonic antigen gene family member 6 (cgm6, cd66b) and nonspecific cross-reacting antigen (nca, cd66c), we prepared their soluble recombinant forms in silkworm larvae. each cdna fragment for cgm6 and nca was ligated into the transfer vector pbk283 after modification to encode the protein lacking the membrane anchor. the resultant vectors were introduced to the bombyx m ...19968776764
green fluorescent protein as a tool for screening recombinant baculoviruses.the gene encoding the green fluorescent protein (gfp) from the jellyfish aequorea victoria, ligated to the honeybee melittin signal peptide-encoding sequence, 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 recombinant green fluorescent protein was identified by sds-page gel electrophoresis followed by coomassie blue staining o ...19968793839
dynamic phosphorylation of autographa californica nuclear polyhedrosis virus pp31.autographa californica nuclear polyhedrosis virus (acmnpv) pp31 is a nuclear phosphoprotein that accumulates in the virogenic stroma, which is the viral replication center in the infected-cell nucleus, binds to dna, and serves as a late expression factor. considering that reversible phosphorylation could influence its functional properties, we examined phosphorylation and dephosphorylation of pp31 in detail. our results showed that pp31 is posttranslationally phosphorylated by both cellular and ...19968794314
initiation of baculovirus dna replication: early promoter regions can function as infection-dependent replicating sequences in a plasmid-based replication assay.from the results of transient plasmid-based replication assays, it has been postulated that homologous regions (hrs) of autographa californica nuclear polyhedrosis virus (acmnpv) function as origins of viral dna replication. however, these assays vary in specificity according to the methodology used and may not be dependent solely on the presence of hr sequences. to determine the role that hrs and other sequences might play in the replication process, a series of plasmids containing specific del ...19968794340
induction of apoptosis by baculovirus transactivator ie1.apoptosis is induced upon infection of sf-21 cells by mutants of the baculovirus autographa californica nuclear polyhedrosis virus (acmnpv) lacking a functional p35 gene which encodes a stoichiometric inhibitor of members of the interleukin-1beta converting enzyme family of cysteine proteases (n.j. bump et al, science 269:1885-1888, 1995; r.j. clem, m. fechheimer, and l.k. miller, science 254:1388-1390, 1991). we found that transfection of sf-21 cells with the acmnpv ie-1 gene was sufficient to ...19968794358
restriction endonuclease analysis of a spodoptera litura nuclear polyhedrosis virus (npv) isolate.the dna of a nuclear polyhedrosis virus (npv) propagated in the larvae of s. litura was analysed with alui, apai, bamhi, bglii, bsteii, bstni, clai, ecori, ecorv, hindiii, hinfi, kpni, mspi, psti, pvuii, sali, sau3ai, smai, taqi, xbai, and xhoi. the unique restriction endonuclease profiles of the virus indicate it to be a distinct npv isolate. the size of the viral genome was estimated to 132 kbp; the genome was mapped with hindiii and psti.19968799322
transcription, translation, and cellular localization of three autographa californica nuclear polyhedrosis virus structural proteins: odv-e18, odv-e35, and odv-ec27.this paper identifies two structural proteins of the occluded derived viral envelope of autographa californica nuclear polyhedrosis virus (acmnpv): odv-e18 and odv-e35. in addition, we identify a protein, odv-ec27, that is incorporated into the capsid of occluded virus, which is not detected in budded virus. the genes for these proteins reside within the ie0 intron. the intron was sequenced, and five open reading frames (orf) were identified. orf 3 (genomic orf 143) codes for the odv envelope pr ...19968806491
cfmnpv blocks acmnpv-induced apoptosis in a continuous midgut cell line.morphological and molecular changes produced by autographa californica nuclear polyhedrosis virus (acmnpv) infection in a permissive cell line, iplb-sf-21ae (sf-21), of spodoptera frugiperda and a nonpermissive cell line, fpmi-cf-203 (cf-203), of choristoneura fumiferana are described. cf-203 cells inoculated with acmnpv showed a dna ladder and morphological changes such as plasma membrane granulation, blebbing, and nuclear fragmentation, which are characteristic of apoptosis. typical virus repl ...19968806500
mapping functional domains in acmnpv pp31.autographa californica nuclear polyhedrosis virus (acmnpv) replicates in the nucleus and produces a viral-modified form of the nuclear matrix called the virogenic stroma. the virogenic stroma is the site of viral dna packaging and nucleocapsid assembly and is thought to be the site of viral dna replication and rna transcription. acmnpv encodes a phosphoprotein, pp31, which localizes to the nucleus of uninfected insect cells and to the virogenic stroma of infected insect cells. pp31 has dna bindi ...19968806516
identification, molecular cloning, and transcription analysis of the choristoneura fumiferana nuclear polyhedrosis virus spindle-like protein gene.the choristoneura fumiferana nuclear polyhedrosis virus spindle-like protein (slp) gene has been identified and localized immediately downstream and in the same orientation as the cfmnpv dna polymerase gene. the slp gene is 1101 bp long, predicted to code for a 366 amino acid (42.1 kda) polypeptide. transcriptional analysis revealed that the cfmnpv slp gene is expressed at late times postinfection, beginning at 24 hr postinfection and is most abundantly expressed after 36 hr. transcription initi ...19968806578
a recombination-efficient baculovirus vector for simultaneous expression of multiple genes.the baculovirus system is an extremely powerful tool for expression of heterologous genes in eukaryotic environment. a multiple expression vector, pbacucmp3, was constructed which harbored two copies of the autographa californica nuclear polyhedrosis virus very late gene promoter and the drosophila melanogaster 70-kda heat-shock protein (hsp70) promoter with downstream unique restriction sites for cloning of three independent foreign genes. co-transfection of pbacucmp3 with bsu36i-linearized vir ...19968666274
the gp64 envelope fusion protein is an essential baculovirus protein required for cell-to-cell transmission of infection.to demonstrate the essential nature of the baculovirus gp64 envelope fusion protein (gp64 efp) and to further examine the role of this protein in infection, we inactivated the gp64 efp gene of autographa californica multicapsid nuclear polyhedrosis virus (acmnpv) and examined the biological properties of this virus in vivo. to provide gp64 efp during construction of the recombinant gp64 efp-null acmnpv baculovirus, we first generated a stably transfected insect cell line (sfpop64-6) that constit ...19968676487
human cathepsin o2, a matrix protein-degrading cysteine protease expressed in osteoclasts. functional expression of human cathepsin o2 in spodoptera frugiperda and characterization of the enzyme.cathepsin o2, a human cysteine protease predominantly present in osteoclasts, has been functionally expressed in spodoptera frugiperda sf9 cells using the autographa californica nuclear polyhedrosis virus. following in vitro activation at ph 4.0 with pepsin, active enzyme with an apparent molecular weight of 29,000 was obtained. n-terminal sequencing revealed the typical processing site for cysteine proteases of the papain family with a proline in the position adjacent to the n-terminal alanine ...19968567669
identification and analysis of an autographa californica nuclear polyhedrosis virus structural protein of the occlusion-derived virus envelope: odv-e56.an autographa californica nuclear polyhedrosis virus gene encoding an occlusion-derived virus (odv) envelope protein of 56 kda was identified and sequenced. transcription initiates from a conserved baculovirus late motif (ataag) with transcripts detected from 16 through 72 hr p.i. the protein is detected in infected cell extracts from 36 hr p.i. western blot assay of odv, bv, viral envelope, and nucleocapsid preparations coupled with immunoelectron microscopy reveal that this protein localizes t ...19968599240
baculovirus replication alters hormone-regulated host development.the baculovirus lymantria dispar nuclear polyhedrosis virus interferes with insect larval development by altering the host's hormonal system. the level of haemolymph ecdysteroids, the insect moulting hormone, was found to be higher in virus-infected larvae than in uninfected controls. this was consistently observed in both fourth instars and day 5-infected fifth instars. the rate of hormone synthesis was examined by in vitro incubation of the prothoracic gland. gland activity in virus-infected l ...19968601794
actin binding and proteolysis by the baculovirus acmnpv: the role of virion-associated v-cath.infection of larvae by autographa californica m nuclear polyhedrosis virus (acmnpv) results in liquefaction of susceptible hosts, presumably due to the breakdown of cells and extracellular matrices. in spodoptera frugiperda tissue culture cells, infection leads to dramatic rearrangement and eventual destruction of the actin cytoskeleton. the first of these rearrangements is the formation of actin cables in the cytoplasm of the cell. cable formation requires release of the budded virus (bv) nucle ...19968607267
baculovirus infection of spodoptera exigua larvae: lacz expression driven by promoters of early genes pe38 and me53 in larval tissue.to follow the progression of infection of autographa californica multicapsid nuclear polyhedrosis virus (acmnpv) within tissues of its larval host, we have constructed acmnpv recombinants carrying lacz reporter genes under the control of the early virus promoters pe38 and me53, in addition to the authentic genes. the early promoter-lacz gene cassettes were located upstream of the very late polyhedrin gene. in infected insect cell lines, pe38 transcription is initiated at an early promoter, while ...19968609477
the dna polymerase and helicase genes of a baculovirus of orgyia pseudosugata.regions of the genome of the orgyia pseudotsugata multinucleocapsid nuclear polyhedrosis virus (opmnpv) containing the dna polymerase and helicase genes were sequenced. the dna polymerase and helicase genes encode predicted proteins of 985 (112.6 kda) and 1223 (140.5 kda) amino acids and exhibited 63 % and 59 % amino acid identity, respectively, with their homologues in the autographa californica mnpv (acmnpv). the influence of sequence variation between the opmnpv and acmnpv dna polymerase and ...19968609478
studies of choristoneura fumiferana nuclear polyhedrosis virus gene expression in insect cells.to investigate the mechanisms regulating baculovirus virulence and host range we have begun to study choristoneura fumiferana nuclear polyhedrosis virus (cfmnpv) and its gene expression in permissive and nonpermissive cells. we have identified and mapped three genes on the cfmnpv genome. the polyhedrin gene is located from 0.0 to 2.0 m.u. and two other genes, dnapol and p143, both of which are essential for baculovirus dna replication, are located from 35.3 to 40.9 m.u. and 55.5 to 63.4 m.u., re ...19968610448
Refresh
firstprev
12345678910...
nextlast
Displaying items 901 - 1000 of 1347