Publications
| Title | Abstract | Year(sorted ascending) Filter | PMID Filter |
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| purification of a virus-induced rna polymerase from autographa californica nuclear polyhedrosis virus-infected spodoptera frugiperda cells that accurately initiates late and very late transcription in vitro. | the virus-induced rna polymerase from autographa californica nuclear polyhedrosis virus-infected spodoptera frugiperda cells was separated from the three host nuclear rna polymerases by deae-sephadex chromatography and then purified through two more steps: heparin-agarose chromatography and glycerol gradient ultracentrifugation. fractions from each of these purification steps have been assayed in vitro for the ability to perform accurate initiation of transcription on a late (p6.9) and a very la ... | 1996 | 8614978 |
| trans-activation of a cell housekeeping gene promoter by the ie1 gene product of baculoviruses. | protein ie1 is the product of a baculovirus gene, ie1, that is activated immediately upon entrance of the viral genome into the cell nucleus. this protein was previously shown to be a trans-regulator of viral genes whose products are required for initiation of the infectious cycle including viral dna replication. to test whether the ie1 protein is also capable of trans-regulating nuclear genes of the host in vitro and in vivo, we transfected the ie1 gene of bombyx mori nuclear polyhedrosis virus ... | 1996 | 8615012 |
| characterization of a highly conserved baculovirus structural protein that is specific for occlusion-derived virions. | a highly conserved baculovirus late gene called odvp-6e was shown to be a structural protein that is specific for occlusion-derived virus (odv) envelopes. the complete sequence of this gene is presented for both orgyia pseudotsugata nuclear polyhedrosis virus (opmnpv) and cydia pomonella granulosis virus (cpgv). the predicted sizes of the opmnpv and cpgv odvp-6e are 40, 241, and 38,655 respectively. the opmnpv odvp-6e gene was transcriptionally mapped and was shown to initiate from a consensus l ... | 1996 | 8615018 |
| the viral ubiquitin gene of autographa californica nuclear polyhedrosis virus is not essential for viral replication. | the baculovirus autographa californica nuclear polyhedrosis virus (acnpv) encodes a protein with significant homology to ubiquitin. to study the role of viral ubiquitin in infection, a recombinant virus was constructed with a frameshift mutation within the coding sequence of the viral ubiquitin gene, v-ubi. this recombinant, named vubi-fs, was viable, indicating that viral ubiquitin is not essential for replication in tissue culture. however, the yields of infectious budded virus were decreased ... | 1996 | 8615029 |
| functional expression of the aequorea victoria green fluorescent protein in insect cells using the baculovirus expression system. | a dna fragment encoding the green fluorescent protein (gfp) was isolated via pcr from a jellyfish aequorea victoria cdna, cloned and sequenced. subsequently, a recombinant baculovirus bearing the coding region of the gfp under the transcriptional control of the autographa californica nuclear polyhedrosis virus (acmnpv) polyhedrin gene promoter was constructed and isolated. high-level expression of gfp could be easily monitored in spodoptera frugiperda (sf9) insect cells after infection with reco ... | 1996 | 8619796 |
| physical map of anagrapha falcifera multinucleocapsid nuclear polyhedrosis virus. | a physical map of anagrapha falcifera multinucleocapsid nuclear polyhedrosis virus (afmnpv) dna was constructed for restriction endonucleases ecori, hindiii, psti and xhoi. the genome size was estimated to be 130 kbp. ordering of the restriction fragments was accomplished by cross-blot hybridization, double digestion and dna-dna hybridization. the polyhedrin gene and homologous repeat (hr) regions were located by hybridization to the autographa californica mnpv (acmnpv) polyhedrin gene and hr4, ... | 1996 | 8627218 |
| baculovirus-mediated gene transfer into mammalian cells. | this paper describes the use of the baculovirus autographa californica multiple nuclear polyhedrosis virus (acmnpv) as a vector for gene delivery into mammalian cells. a modified acmnpv virus was prepared that carried the escherichia coli lacz reporter gene under control of the rous sarcoma virus promoter and mammalian rna processing signals. this modified baculovirus was then used to infect a variety of mammalian cell lines. after infection of the human liver cell lines hepg2, >25% of the cells ... | 1996 | 8637876 |
| human bleomycin hydrolase: molecular cloning, sequencing, functional expression, and enzymatic characterization. | we have cloned the cdna of human bleomycin hydrolase (hbh), a protease which is thought to be involved in the metabolic inactivation of the antineoplastic drug bleomycin. the open reading frame consists of 1365 base pairs and is predicted to encode a 52 kda protein. the protein shares 40% identity with yeast bleomycin hydrolase and contains the conserved active site residues (cys, his, asn) characteristic for cysteine proteases of the papain superfamily. human bleomycin hydrolase has been functi ... | 1996 | 8639621 |
| cloning and biologic activities of a bovine interferon-alpha isolated from the epithelium of a rotavirus-infected calf. | a cdna encoding a distinct bovine (bo) interferon (ifn) alpha, designated boifn-alpha e, was generated from gut epithelial cells isolated from a rotavirus-infected calf. the boifn-alpha e cdna sequence shared a greater than 90% identity with the other boifn-alpha subtypes. the cdna encoding boifn-alpha e has been expressed in insect cells using the baculovirus autographa californica nuclear polyhedrosis virus (acmnpv) as a vector. insect cells infected with recombinant virus secreted a protein w ... | 1996 | 8640447 |
| identification of baculovirus gene that promotes autographa californica nuclear polyhedrosis virus replication in a nonpermissive insect cell line. | a gene that promotes autographa californica m nuclear polyhedrosis virus (acmnpv) replication in iplb-ld652y cells, a cell line that is nonpermissive for acmnpv, was identified in lymantria dispar m nuclear polyhedrosis virus (ldmnpv). cotransfection of acmnpv dna and a plasmid carrying the ldmnpv gene into iplb-ld652y cells results in acmnpv replication. the gene maps between 43.3 and 43.8 map units on the 162-kbp genome of ldmnpv. it comprises a 218-codon open reading frame and encodes a polyp ... | 1996 | 8642646 |
| factors regulating baculovirus late and very late gene expression in transient-expression assays. | eighteen genes of autographa californica nuclear polyhedrosis virus are necessary and sufficient to transactivate expression from the late vp39 promoter in transient-expression assays in sf-21 cells. these 18 genes, known as late expression factor genes (lefs), are also required to transactivate the very late promoter of the polyhedrin gene, polh, but expression from this promoter is relatively weak compared with expression from the vp39 promoter. to further define the factors required for late ... | 1996 | 8642657 |
| cloning and expression of apoptosis inhibitory protein homologs that function to inhibit apoptosis and/or bind tumor necrosis factor receptor-associated factors. | baculovirus inhibitors of apoptosis (iaps) act in insect cells to prevent cell death. here we describe three mammalian homologs of iap, miha, mihb, and mihc, and a drosophila iap homolog, diha. each protein bears three baculovirus iap repeats and an n-terminal ring finger motif. apoptosis mediated by interleukin 1beta converting enzyme (ice), which can be inhibited by orgyia pseudotsugata nuclear polyhedrosis virus iap (opiap) and cowpox virus crma, was also inhibited by miha and mihb. as mihb a ... | 1996 | 8643514 |
| a new set of versatile vectors for the heterologous expression of foreign genes using the baculovirus system. | a new set of versatile advanced baculovirus (bv) vectors for the production of fused proteins in insect cells, under the control of the strong polyhedrin promoter of the autographa california nuclear polyhedrosis virus (acmnpv), has been constructed. the vectors contain peptide tags which allow immunological detection, as well as purification of recombinant protein produced via the bv expression system. | 1996 | 8647444 |
| replication of simian virus 40 origin-containing dna during infection with a recombinant autographa californica multiple nuclear polyhedrosis virus expressing large t antigen. | autographica californica multiple nuclear polyhedrosis virus (acmnpv) has been shown to encode many of the enzymes involved in the replication of its own dna. although the acmnpv genome contains multiple sets of reiterated sequences that are thought to function as origins of dna replication, no initiator protein has yet been identified in the set of viral replication enzymes. in this study, the ability of a heterologous origin initiator system to promote dna replication in acmnpv-infected cells ... | 1997 | 8985377 |
| expression of foreign proteins on the surface of autographa californica nuclear polyhedrosis virus. | based on the method of direct cloning into the baculovirus genome by linearizing and re-ligation in presence of the target insert, we designed viral constructs that express foreign genes on the surface of baculovirus particles. we chose the glycosylated envelope protein gp41 of human immunodeficiency virus type 1 (hiv-1) as a model for displaying recombinant proteins on budded virus. the ectodomain of the envelope protein gp41 of hiv-1 was being fused to the entire baculovirus major coat protein ... | 1997 | 9105625 |
| n-terminal sequences from autographa californica nuclear polyhedrosis virus envelope proteins odv-e66 and odv-e25 are sufficient to direct reporter proteins to the nuclear envelope, intranuclear microvesicles and the envelope of occlusion derived virus. | baculovirus occlusion-derived virus (odv) derives its envelope from an intranuclear membrane source. n-terminal amino acid sequences of the autographa californica nuclear polyhedrosis virus (acmnpv) envelope proteins, odv-e66 and odv-e25 (23 and 24 amino acids, respectively) are highly hydrophobic. recombinant viruses that express the two n-terminal amino acid sequences fused to green fluorescent protein (23gfp or 24gfp) provided visual markers to follow protein transport and localization within ... | 1997 | 9108103 |
| organization and molecular characterization of genes in the polyhedrin region of the anagrapha falcifera multinucleocapsid npv. | a multinucleocapsid nuclear polyhedrosis virus (mnpv) isolated from the celery looper, anagrapha falcifera, has been proposed as a new virus based on differences in virulence and dna fragment profiles between this isolate and the autographa californica mnpv, the mnpv type species. in the present study, we examined the relatedness of the afmnpv and acmnpv genomes by (1) southern hybridization, (2) comparison of their genetic organization in the polyhedrin gene region (acmnpv ecori-i fragment), an ... | 1997 | 9125047 |
| the sequence of the orgyia pseudotsugata multinucleocapsid nuclear polyhedrosis virus genome. | the nucleotide sequence of the orgyia pseudotsugata multinucleocapsid nuclear polyhedrosis virus (opmnpv) genome was completed and analyzed. it is composed of 131,990 bases with a g + c content of 55% and contains 152 putative genes of 150 nucleotides or greater. major differences in gene content and arrangement between opmnpv and the autographa californica mnpv were found. these include the presence in opmnpv of three complete iap gene homologs, two conotoxin gene homologs, two protein tyrosine ... | 1997 | 9126251 |
| deletion analysis of four of eighteen late gene expression factor gene homologues of the baculovirus, bmnpv. | nucleotide sequence analysis of the genome of the baculovirus bombyx mori nuclear polyhedrosis virus (bmnpv) identified 18 homologues of the autographa californica npv (acnpv) lefs (late expression factor genes). these bmnpv lefs showed high (73-98%) amino acid sequence identities to acnpv lefs and were localized to similar positions in the genome. one lef, p35, was previously characterized in acnpv and bmnpv deletion experiments. functional deletion of each of the bmnpv lef homologues was attem ... | 1997 | 9126260 |
| baculovirus-mediated trans-epithelial transport of proteins in infected caterpillars. | baculovirus-mediated abnormal protein trafficking was studied in infected caterpillars by using heterologous proteins. the gene for human complement c1r was expressed in larvae of mamestra brassicae by a recombinant autographa californica nuclear polyhedrosis virus (acmnpv) vector. by following the time-course of recombinant c1r distribution among various tissues, cell types and cell organelles, we concluded that the dominant site of recombinant protein synthesis was the fat body, although some ... | 1997 | 9129671 |
| baculovirus inhibitor of apoptosis functions at or upstream of the apoptotic suppressor p35 to prevent programmed cell death. | members of the inhibitor of apoptosis (iap) gene family prevent programmed cell death induced by multiple signals in diverse organisms, suggesting that they act at a conserved step in the apoptotic pathway. to investigate the molecular mechanism of iap function, we expressed epitope-tagged op-iap, the prototype viral iap from orgyia pseudotsugata nuclear polyhedrosis virus, by using novel baculovirus recombinants and stably transfected insect cell lines. epitope-tagged op-iap blocked both virus- ... | 1997 | 9151843 |
| late promoter selection in the baculovirus gp64 envelope fusion protein gene. | the upstream promoter region of the autographa californica multicapsid nuclear polyhedrosis virus (acmnpv) gp64 gene contains five copies of taag, the conserved sequence found at the transcriptional initiation sites of almost all baculovirus late genes. in acmnpv-infected sf9 cells, late transcription initiation is detected from only two upstream taag sites and not from three downstream taag sites. to examine several models for preferential taag site utilization, we constructed a series of recom ... | 1997 | 9168879 |
| loading of iron into recombinant rat liver ferritin heteropolymers by ceruloplasmin. | we have reported previously that the heavy chain of ferritin is required for iron incorporation by ceruloplasmin (j.-h. guo, m. abedi, and s. d. aust (1996) arch. biochem. biophys. 335(1)). the purpose of this study was to determine how many heavy chains were required for ceruloplasmin to interact with ferritin such that iron loading occurred. the cdna sequences encoding the heavy and light chains of rat liver ferritin were cloned into the baculovirus transfer vector pa-cuw51 under the control o ... | 1997 | 9169016 |
| [transgenerational transfer of gypsy moth (ocneria dispar l.) nuclear polyhedrosis virus]. | gypsy moth nuclear polyhedrosis virus infects egg laying under natural conditions. the virus was identified by restriction endonucleases pst1, bamh1, and sal1. | 1997 | 9182408 |
| expression of lacz reporter gene under the control of the polyhedrin promoter of spodoptera litura nuclear polyhedrosis virus. | promoter function of the putative polyhedrin-encoding gene (polh) of spodoptera litura nuclear polyhedrosis virus (s1mnpv) was determined by transferring it to the autographa californica nuclear polyhedrosis virus (acmnpv) through the acnpv polh based vector, pvl1393. three transfer vectors pcbt2, pcbt3 and pcbt4 were constructed by substituting the promoter and the neighbouring sequences of acnpv in pvl1393 by that of s1npv. the escherichia coli lacz gene was placed downstream from the s1npv po ... | 1997 | 9185860 |
| sequencing of the putative dna helicase-encoding gene of the bombyx mori nuclear polyhedrosis virus and fine-mapping of a region involved in host range expansion. | the complete sequence of a 3666-nucleotide (nt) open reading frame (orf) and its flanking regions (58.1-62.1 map units (m.u.) from bombyx mori nuclear polyhedrosis virus (bmnpv)) was determined. this orf, bmnpv dnahel, encoded a predicted protein of 143623 da which possessed seven consensus motifs found in proteins which unwind duplex dnas, indicating that it is a dna helicase. a 572-bp saci-hindiii fragment, bmsch, that was previously shown to expand the host range of autographa californica npv ... | 1997 | 9185864 |
| heterologous promoter recognition leading to high-level expression of cloned foreign genes in bombyx mori cell lines and larvae. | the baculovirus expression system using the autographa californica nuclear polyhedrosis virus (acnpv) has been extensively utilized for high-level expression of cloned foreign genes, driven by the strong viral promoters of polyhedrin (polh) and p10 encoding genes. a parallel system using bombyx mori nuclear polyhedrosis virus (bmnpv) is much less exploited because the choice and variety of bmnpv-based transfer vectors are limited. using a transient expression assay, we have demonstrated here tha ... | 1997 | 9185865 |
| identification and characterization of a baculovirus structural protein, vp1054, required for nucleocapsid formation. | the defect in a temperature-sensitive mutant of autographa californica nuclear polyhedrosis virus, tsn1054, was mapped and characterized. at the nonpermissive temperature of 33 degrees c, this mutant fails to form plaques upon infection of sf-21 cultured insect cells; infection is limited to a single cell, even though the infection proceeds through the very late phase. marker rescue mapping and dna sequencing identified the gene, orf 54, which was altered by a single nucleotide substitution in t ... | 1997 | 9188569 |
| a mechanism for negative gene regulation in autographa californica multinucleocapsid nuclear polyhedrosis virus. | the autographa californica multinucleocapsid nuclear polyhedrosis virus (acmnpv) ie-1 gene product (ie-1) is thought to play a central role in stimulating early viral transcription. ie-1 has been demonstrated to activate several early viral gene promoters and to negatively regulate the promoters of two other acmnpv regulatory genes, ie-0 and ie-2. our results indicate that ie-1 negatively regulates the expression of certain genes by binding directly, or as part of a complex, to promoter regions ... | 1997 | 9188574 |
| dna replication promotes high-frequency homologous recombination during autographa californica multiple nuclear polyhedrosis virus infection. | the relative ease with which foreign genes can be incorporated into the genome of the baculovirus autographa californica nuclear polyhedrosis virus (acmnpv) indicates that a highly efficient recombinational process exists within infected cells. however, it is unclear whether this is due to marker transfer mediated by host cell enzymes or recombination events promoted by acmnpv itself. to address the latter possibility, a pair of inverted repeat is50 elements derived from the bacterial transposon ... | 1997 | 9191843 |
| baculovirus transactivator ie1 is functional in mammalian cells. | ie1 of autographa californica multicapsid nuclear polyhedrosis virus acts as a transactivator of several viral promoters in insect cells. transient expression assays indicate that ie1 is involved in the activation of the early promoter he65 in both insect tn-368 and mammalian bhk-21 cell lines. ie1 activation of the he65 promoter was compared with ie1 activation of the early 39k promoter. in contrast to the he65 promoter, the 39k promoter was not inducible by ie1 in bhk-21 cells. | 1997 | 9191949 |
| characterization of p91, a protein associated with virions of an orgyia pseudotsugata baculovirus. | polyclonal antiserum produced against preoccluded virions from the orgyia pseudotsugata multinucleocapsid nuclear polyhedrosis virus (opmnpv) was used to screen an opmnpv lambda gt11 expression library. the insert from one of the immunoreactive phage isolates hybridized to opmnpv orf86 (p91), a 2460-bp (819 amino acids) open reading frame that encodes a predicted protein of 91 kda. antibodies generated against a maltose binding protein-p91 fusion detected a band of approximately 91 kda on wester ... | 1997 | 9201231 |
| differential requirements of two insect cell lines for growth in serum-free medium. | the development of a serum-free medium that supports the growth of cells from a spodoptera frugiperda and a lymantria dispar cell line is reported. a yeast hydrolysate provided the b-vitamin complex, and a combination of a meat hydrolysate and tryptose provided most of the free amino acids required for cell growth. supplemental cystine and methionine were required to achieve maximum cell growth. the serum or serum replacements used in earlier formulations were replaced with commercial lipid prep ... | 1997 | 9201517 |
| association of a baculovirus-encoded protein with the capsid basal region. | an open reading frame homologous to acmnpv orf9 (orf1629) was characterized in the orgyia pseudotsugata multinucleocapsid nuclear polyhedrosis virus (opmnpv). sequence analysis indicated that the opmnpv homolog (called orf2) encoded a protein predicted to contain 545 amino acids with a molecular weight of 61 kda. the first 80 amino acids did not have a counterpart in the acmnpv homolog. the remainder of the orf was poorly conserved with 29% amino acid identity overall with the acmnpv orf. howeve ... | 1997 | 9007067 |
| detection and identification of multiple baculoviruses using the polymerase chain reaction (pcr) and restriction endonuclease analysis. | a technique using the polymerase chain reaction (pcr) and restriction analysis was developed for the simultaneous detection of eight baculoviruses. the baculoviruses detected by this technique were autographa californica multiple-embedded nuclear polyhedrosis virus (mnpv). anticarsia gemmatalis mnpv, bombyx mori mnpv, orgyia pseudotsugata mnpv. spodoptera frugiperda mnpv, s. exigua mnpv, anagrapha falcifera mnpv, heliothis zea single-embedded nuclear polyhedrosis virus (snpv). a highly conserved ... | 1997 | 9015292 |
| autographa californica nuclear polyhedrosis virus p143 gene product is a dna-binding protein. | we have identified the protein product of the autographa californica nuclear polyhedrosis virus (acmnpv) p143 gene by constructing a recombinant baculovirus overexpressing the gene product p143. the overexpressed protein exhibited a relative mobility of approximately 140 kda and was stable for at least 12 hr after synthesis. immunoblotting using a monoclonal antibody developed against the overexpressed protein identified a similar polypeptide in acmnpv-infected cells which was detectable by 4 hr ... | 1997 | 9024814 |
| establishment of a cell line from spodoptera litura (lepidoptera: noctuidae) and replication of s. litura nuclear polyhedrosis virus in vitro. | a new cell line, designated ibl-slo1a, was established from pupal ovaries of the tobacco cutworm spodoptera litura. cells were grown in the tnm-fh insect cell culture medium supplemented with 10% fetal bovine serum. the subculture of cells was initiated with 1-3 x 10(5) cells/ml, a split ratio about 1:10. the growth curve of the cells fit the model of exponential growth (y = e-0.6750 + 0.0317x r2 = 0. 98), the population doubling time during logarithmic growth at 28 degrees c was 21.9 hr. the nu ... | 1997 | 9028921 |
| replication of anticarsia gemmatalis nuclear polyhedrosis virus in four lepidopteran cell lines. | anticarsia gemmatalis nuclear polyhedrosis virus (agnpv; family baculoviridae) is pathogenic for larvae of the velvetbean caterpillar, anticarsia gemmatalis hübner-an important pest of soybean. agnpv is a viable alternative to chemical control of a. gemmatalis in brazil, where its use as a pesticide has brought significant economic and environmental benefits. although a significant amount of information is available on the ecological and biological control aspects of agnpv, very little is known ... | 1997 | 9028926 |
| characterization of a domain of the genome of bmnpv containing a functional gene for a small capsid protein and harboring deletions eliminating three open reading frames that are present in acnpv. | the nucleotide sequence of a 1.5-kb fragment of the genome of bombyx mori nuclear polyhedrosis virus (bmnpv) has been determined. an open reading frame (orf) has been identified, which encodes a protein of a predicted molecular mass of 15 kda (p15). analysis of the p15 sequence revealed that it shares significant similarities with a number of previously characterized viral capsid proteins. two mrnas, 0.7- and 1.3-kb, are transcribed from the p15 gene. the 0.7-kb transcript appears before bmnpv d ... | 1997 | 9034315 |
| expression of the mouse interleukin-2 receptor gamma chain in insect cells using a baculovirus expression vector--comparison with the human common gamma chain. | the gene encoding the gamma-chain of the mouse interleukin-2 receptor was expressed in lepidopteran insect cells using the baculovirus expression vector system. the corresponding gene was inserted under 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 receptor protein was identified by immunoblotting in cell lysates prepared from insect cells infected with the p ... | 1997 | 9042425 |
| a role for baculovirus gp41 in budded virus production. | insect cells infected with tsb1074, a temperature-sensitive mutant of autographa californica nuclear polyhedrosis virus, exhibit a "single-cell-infection" phenotype whereby the infection progresses through the very late phase culminating in occlusion body formation, but neighboring cells do not become infected. marker rescue mapping and dna sequencing correlated a single nucleotide substitution within the baculovirus gp41 gene with the temperature-sensitive phenotype of tsb1074. the product of t ... | 1997 | 9217053 |
| complete sequence and transposon mutagenesis of the bamhi j fragment of cydia pomonella granulosis virus. | the bamhi j fragment of cydia pomonella granulosis virus was subcloned and subjected to transposon mutagenesis in escherichia coli using a tn3 derivative. after screening by restriction endonuclease digestion and polymerase chain reaction (pcr), 44 clones were selected representing insertions every 100 to 300 bp. the complete sequence was compiled and the transposon insertion sites mapped precisely by sequencing. analysis of the sequence revealed the presence of 7 potential open reading frames ( ... | 1997 | 9237352 |
| expression of neuroparsin cdna in insect cells using baculovirus vectors. | the cdna encoding neuroparsin a, a polytropic neurohormone of the locust, locusta migratoria, was inserted into the genome of autographa californica nuclear polyhedrosis virus such that transcription was under control of the p10 promoter. a polypeptide having the same charge and the same apparent molecular weight as the authentic neuroparsin a and that was reactive against neuroparsin immune serum was produced in recombinant virus-infected lepidopteran cell lines but not in control virus-infecte ... | 1997 | 9243790 |
| regulation of infected-cell-specific protein synthesis in sf iplb-21 cells productively infected with spodoptera frugiperda multicapsid nuclear polyhedrosis virus. | in spodoptera frugiperda (sf iplb-21) cells productively infected with s. frugiperda multicapsid nuclear polyhedrosis virus (sfmnpv), we observed the synthesis of infected-cell-specific polypeptides (icsps) using the protein synthesis inhibitor cycloheximide (cx) and the dan synthesis inhibitor cytosine arabinoside (ara-c) in an attempt to assess whether a temporal cascade and viral dna synthesis are involved in the gene expression program of sfmnpv. inhibition of protein synthesis with cx at 0 ... | 1997 | 9268770 |
| a nuclear polyhedrosis virus from the obliquebanded leafroller, choristoneura rosaceana (harris) (lepidoptera:tortricidae). | a nuclear polyhedrosis virus (crmnpv) has been isolated from larvae of the obliquebanded leafroller choristoneura rosaceana (harris) collected near st. quentin in northwest new brunswick, canada. restriction endonuclease digestion-gel electrophoresis of this virus, using a number of different endonucleases and whole crmnpv genome southern hybridization, have shown it to be different but similar to the equivalent virus from the spruce budworm choristoneura fumiferana (clem.) (cfmnpv). the polyhed ... | 1997 | 9281399 |
| ingestion time does not influence the susceptibility of trichoplusia ni to a nuclear polyhedrosis virus | 1997 | 9281407 | |
| baculovirus expression of a human g2-specific, neutralizing igg monoclonal antibody to puumala virus. | we amplified by polymerase chain reaction the heavy- and light-chain antibody genes of a human hybridoma secreting a neutralizing, igg monoclonal antibody to the g2 protein of puumala virus. the heavy- and light-chain genes were cloned and sequenced and the deduced amino acids were aligned with those of other human antibodies to identify the constant and variable regions. the genes were cloned into the baculovirus plasmid transfer vector pacuw51 such that the heavy-chain and light-chain genes we ... | 1997 | 9281505 |
| stable transformation of insect cells to coexpress a rapidly selectable marker gene and an inhibitor of apoptosis. | we have constructed several plasmid expression vectors to express foreign genes in stably transformed insect cells. unlike baculovirus-based expression vectors by which genes of interest are expressed transiently before lysis of the virus-infected cells, genes can be expressed continuously over many passages in a stable cell line. furthermore, the function of a gene or genes expressed in a stable cell line from an insect-specific promoter that is constitutively expressed can be studied in the ab ... | 1997 | 9282319 |
| splicing is required for transactivation by the immediate early gene 1 of the lymantria dispar multinucleocapsid nuclear polyhedrosis virus. | a region of the lymantria disper multinucleocapsid nuclear polyhedrosis virus (ldmnpv) genome containing the homolog of the baculovirus ie-1 gene was identified using a series of overlapping cosmids and individual plasmids in a transient transcriptional expression assay. sequence analysis of the active region identified two orfs, one of which is 32% identical to acmnpv orf141 (ie-0) and contains a putative splice donor site and the other of which is 29% identical to acmnpv ie-1 and contains a hi ... | 1997 | 9300047 |
| characterization of the polyhedrin gene of spodoptera litura nuclear polyhedrosis virus. | the polyhedrin gene (polh) of a characteristically distinct spodoptera litura nuclear polyhedrosis virus isolate (simnpv) is identified in the hindiii-f fragment of the viral dna. the nucleotide sequence of the 1057 base pair (bp) region of this fragment contains an open reading frame (orf) without any intervening sequence for coding a polypeptide of 246 amino acids. analysis of the nucleotide sequence and deduced amino acid sequence indicate that this has more than 70% sequence identity to know ... | 1997 | 9311561 |
| baculovirus gp64 gene expression: negative regulation by a minicistron. | small upstream open reading frames (orfs) or minicistrons located in the 5' leader of eukaryotic mrnas have been shown to play a role in translational regulation of some eukaryotic genes, particularly mammalian proto-oncogenes. a survey of the baculovirus autographa californica multicapsid nuclear polyhedrosis virus genome suggests that at least 10 transcripts from late genes contain potential minicistrons, and at least three of these minicistrons appear to be conserved in homologous genes of th ... | 1997 | 9311824 |
| expression of the autographa californica nuclear polyhedrosis virus apoptotic suppressor gene p35 in nonpermissive spodoptera littoralis cells. | apoptosis was postulated as the main barrier to replication of the autographa californica nuclear polyhedrosis virus (acmnpv) in a spodoptera littoralis sl2 cell line (n. chejanovsky and e. gershburg, virology 209:519-525, 1995). thus, we hypothesized that the viral apoptotic suppressor gene p35 is either poorly expressed or nonfunctional in acmnpv-infected sl2 cells. these questions were addressed by first determining the steady-state levels of the p35 product, p35, in acmnpv-infected sl2 cells ... | 1997 | 9311840 |
| responses of insect cells to baculovirus infection: protein synthesis shutdown and apoptosis. | protein synthesis is globally shut down at late times postinfection in the baculovirus autographa californica m nuclear polyhedrosis virus (acmnpv)-infected gypsy moth cell line ld652y. a single gene, hrf-1, from another baculovirus, lymantria dispar m nucleopolyhedrovirus, is able to preclude protein synthesis shutdown and ensure production of acmnpv progeny in ld652y cells (s. m. thiem, x. du, m. e. quentin, and m. m. berner, j. virol. 70:2221-2229, 1996; x. du and s. m. thiem, virology 227:42 ... | 1997 | 9311875 |
| identification of the early actin-rearrangement-inducing factor gene, arif-1, from autographa californica multicapsid nuclear polyhedrosis virus. | in response to autographa californica multicapsid nuclear polyhedrosis virus (acmnpv) infection, a sequential rearrangement of the actin cytoskeleton occurs. previous studies suggest that the penetration of nucleocapsids induces early actin cables followed by further changes of the actin cytoskeleton which depend on early viral gene expression. by transfection of a plasmid library into trichoplusia ni tn-368 cells, we have identified an early viral gene, designated arif-1, that is able to induce ... | 1997 | 9311884 |
| baculoviral display of the green fluorescent protein and rubella virus envelope proteins. | the ability to display heterologous proteins and peptides on the surface of different types of bacteriophage has proven extremely useful in protein structure/function studies. to display such proteins in a eucaryotic environment, we have produced a vector allowing for fusion of proteins to the amino-terminus of the autographa californica nuclear polyhedrosis virus (acnpv) major envelope glycoprotein, gp64. such fusion proteins incorporate into the baculoviral virion and display the flag epitope ... | 1997 | 9325155 |
| molecular analysis of an enhancin gene in the lymantria dispar nuclear polyhedrosis virus. | a lymantria dispar nuclear polyhedrosis virus (ldmnpv) gene has been identified that encodes a homolog to the granulovirus (gv) enhancin proteins that are capable of enhancing the infection of other baculoviruses. enhancin genes have been identified and sequenced for three species of gvs but have not been found in any other nuclear polyhedrosis virus to date. the ldmnpv enhancin gene is located between 67.6 and 70.1 kbp on the viral genome. northern and primer extension analyses of viral rnas in ... | 1997 | 9343163 |
| infectivity and effects of gypsy moth and spruce budworm nuclear polyhedrosis viruses ingested by rainbow trout. | rainbow trout fingerlings were fed dried krill injected with gypsy moth or spruce budworm nuclear polyhedrosis virus (ldnpv and cfnpv, respectively) at a total dose of 1.4 x 10(7) occlusion bodies (obs) per fish. by the end of the 21-day experimental period there were no adverse effects on fish survival or behavior and no significant differences in feeding rates or growth between treated and control fish. the internal organs of all fish were examined at the end of the experiment and there were n ... | 1997 | 9352215 |
| production of recombinant herpes simplex virus protease in 10-l stirred vessels using a baculovirus-insect cell expression system. | a gene expression system using recombinant autographa california nuclear polyhedrosis virus (baculovirus) and sf-9 cells has been scaled up to the 10-l tank level and shown to be capable of producing herpes simplex virus (hsv) protease in serum-free media. high densities of spodoptera frugiperda (sf-9) cells were achieved by modifying two 10-l biolafitte fermenters specifically for insect cell growth. the existing rushton impellers were replaced by marine impellers to reduce shear and the aerati ... | 1997 | 9366090 |
| dna-dependent transregulation by ie1 of autographa californica nuclear polyhedrosis virus: ie1 domains required for transactivation and dna binding. | ie1 is the principal early transregulator of autographa californica multicapsid nuclear polyhedrosis virus (acmnpv). the 582-residue protein stimulates viral transcription and binds as a dimer to 28-bp palindromic repeats (28-mers) comprising the acmnpv homologous region (hr) transcription enhancers. to define ie1 domains responsible for hr-dependent transactivation, we first constructed a series of ie1 fusions to the dna binding domain of the yeast gal4 transactivator. in transfection assays, g ... | 1997 | 9371585 |
| identification of two independent transcriptional activation domains in the autographa californica multicapsid nuclear polyhedrosis virus ie1 protein. | the autographa californica multicapsid nuclear polyhedrosis virus immediate-early protein, ie1, is a 582-amino-acid phosphoprotein that regulates the transcription of early viral genes. deletion of n-terminal regions of ie1 in previous studies (g. r. kovacs, j. choi, l. a. guarino, and m. d. summers, j. virol. 66:7429-7437, 1992) resulted in the loss of transcriptional activation, suggesting that this region may contain an acidic activation domain. to identify independently functional transcript ... | 1997 | 9371622 |
| interactions of recombinant and wild-type baculoviruses with classical insecticides and pyrethroid-resistant tobacco budworm (lepidoptera: noctuidae). | in tests with neonate heliothis virescens (f.), we characterized interactions of all combinations of a recombinant autographa californica (speyer) nuclear polyhedrosis virus (acaait) that expresses an insect-selective neurotoxin (aait) and wild-type acnpv when combined with low concentrations of several conventional insecticides. all combinations of the recombinant virus acaait and insecticides showed a positive interaction (decrease in the median lethal time (lt50) compared with the lt50 for ei ... | 1997 | 9374595 |
| characterization of the replication cycle of the lymantria dispar nuclear polyhedrosis virus. | the life cycle of the lymantria dispar nuclear polyhedrosis virus (ldmnpv) was characterized through analysis of budded virus (bv) release, the temporal formation of polyhedra, the temporal transcription pattern of representative early, late, and hyper-expressed late genes, and the onset of dna replication in the ld652y cell line. transcripts from the ldmnpv immediate early gene g22 were detected 4 h post infection (h p.i.). the late and hyper-expressed late p39 capsid and polyhedrin genes were ... | 1997 | 9381798 |
| a baculovirus (bombyx mori nuclear polyhedrosis virus) repeat element functions as a powerful constitutive enhancer in transfected insect cells. | it has been previously reported that baculovirus homologous regions, the regions of baculovirus genomes that contain the origins of dna replication, can augment the expression of a small number of baculovirus genes in vitro. we are now reporting that a region of the genome of bombyx mori nuclear polyhedrosis virus (bmnpv) containing the homologous region 3 (hr3) acts as an enhancer for the promoter of a nonviral gene, the cytoplasmic actin gene of the silkmoth b. mori. incorporation of the hr3 s ... | 1997 | 9388209 |
| efficient protein production using a bombyx mori nuclear polyhedrosis virus lacking the cysteine proteinase gene. | infection by a baculovirus (bombyx mori nuclear polyhedrosis virus, bmnpv) in silkworm (bombyx mori) larvae is highly efficient as an expression system for the production of useful proteins. however, the amount of the protein of interest expressed tends to decrease in the later stages of infection presumably due, in part, to a proteinase produced in the larval haemolymph. the n-terminal amino acid sequence of a proteinase purified from the haemolymph of bmnpv-infected larvae was identical to the ... | 1997 | 9400955 |
| expression of human immunodeficiency virus genes using baculovirus expression system. | the structural protein genes of hiv-1 and hiv-2 have been expressed in spodoptera frugiperda (sf) cells using baculovirus expression system. the noncoding flanking sequences of hiv structural genes were removed and a putative ribosome binding site was placed in front of the open reading frame of each gene by using crossover linker mutagenesis. the coding sequences of the gag, pol, env, and vif proteins were inserted into autographa californica nuclear polyhedrosis virus (acnpv) so that hiv genes ... | 1997 | 9406188 |
| detection of antibodies against porcine parvovirus nonstructural protein ns1 may distinguish between vaccinated and infected pigs. | the humoral antibody response against the nonstructural protein ns1 and the structural protein vp2 of porcine parvovirus (ppv) was evaluated by immuno-peroxidase test (ipt) and enzyme linked immuno sorbent assay (elisa) using recombinant ppv antigens. the coding sequence for ns1 and vp2 was inserted into the baculovirus. autographa californica nuclear polyhedrosis virus (acnpv) genome resulting in two recombinant baculoviruses acnpv-ns1 and acnpv-vp2, respectively. sf9 cells (spodoptora frugidip ... | 1997 | 9050166 |
| enhanced bioactivity of recombinant baculoviruses expressing insect-specific spider toxins in lepidopteran crop pests | two genetically enhanced isolates of the autographa californica nuclear polyhedrosis virus (acmnpv) expressing insect-specific neurotoxin genes from the spiders diguetia canities and tegenaria agrestis were evaluated for their commercial potential. since prevention of feeding damage is of primary importance in assessing agronomic efficacy, a method for estimating the median time to cessation of feeding (ft50) was developed. neonate droplet feeding assays with preoccluded virus samples were condu ... | 1997 | 9056461 |
| microbial contamination in spodoptera littoralis nuclear polyhedrosis virus produced in insects in egypt | spodoptera littoralis nuclear polyhedrosis virus produced in egypt, for use in field trials for the control of s. littoralis on cotton, was subject to microbiological examination to quantify microbial contamination. bacteria were found to be present at 10(6)-10(9) colony-forming units/ml in virus suspensions containing 2.1 x 10(9) polyhedral inclusion bodies/ml. batches were found to contain between 2 and 11 different species of contaminant microbes. no primary human pathogens of medical importa ... | 1997 | 9056465 |
| polyomavirus major capsid protein vp1 is capable of packaging cellular dna when expressed in the baculovirus system. | using the p2bac dual multiple cloning site transfer vector, the polyomavirus major capsid protein gene vp1 was cloned for expression in the baculovirus-insect cell expression system. the 5-day-infected cellular lysate from this recombinant preparation was purified by cesium chloride density gradient centrifugation. capsid-like particles were observed in the resulting preparation. the purified particle preparation was analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and was s ... | 1997 | 9060642 |
| characterization of the interaction between the baculovirus replication factors lef-1 and lef-2. | the autographa californica multinucleocapsid nuclear polyhedrosis virus has six genes required and three genes stimulatory for transient dna replication. we demonstrate that the products of two of these genes, lef-1 and lef-2, interact in both two-hybrid assays using saccharomyces cerevisiae and glutathione s-transferase fusion affinity assays. using yeast-two-hybrid assays, we mapped the interaction domain of lef-2 to amino acids between positions 20 and 60. extensive deletion analyses of lef-1 ... | 1997 | 9060674 |
| expression of rat steroid 5 alpha-reductase (isozyme-1) in spodoptera frugiperda, sf21, insect cells: expression of rat steroid 5 alpha-reductase. | the enzyme steroid 5 alpha-reductase (5 alpha r) catalyzes the reduction of testosterone (t) to 5 alpha-dihydrotestosterone (dht). in this study, the baculovirus expression system was used to overexpress rat 5 alpha r type i isozyme (r5 alpha r 1). the full length of r5 alpha r1 cdna was inserted into the autographa californica nuclear polyhedrosis virus (ac-mnpv) genome and expressed in spodoptera frugiperda, sf 21, insect cells. the expressed recombinant r5 alpha-r1 showed maximal enzymatic ac ... | 1997 | 9090798 |
| [comparative characteristics of nuclear polyhedrosis virus isolates from gypsy moth lymantria dispar l]. | eight strains of gypsy moth nuclear polyhedrosis virus (npv) were isolated from dead larvae in kirghizia and tyumen and novosibirsk districts. the strains were characterized in terms of polyhedron morphology, biological activity, virus production, and restriction analysis of dna. the isolated strains did not differ in size or shape of polyhedrons. npv strain nshn-2 was characterized by the highest biological activity (ld50 = 1.2 +/- 0.4) x 10(4) and maximum production: 7.3 x 10(8) pib/larva in c ... | 1997 | 9103045 |
| characterization of recombinant autographa californica nuclear polyhedrosis virus (npv) expressing the beta-galactosidase gene in both sf21 and bm5 cells by bombyx mori npv p143 helicase gene. | genomic dna of recombinant acnpv expressing beta-galactosidase was cotransfected with p143 helicase gene of bmnpv into sf21 cells. ac-bm hybrid viruses capable of replicating in both bm5 and sf21 cells were isolated. ac-bm hybrid viruses expressing beta-galactosidase either at the highest (ac-bm hybrid virus-he) or lowest (ac-bm hybrid virus-le) level were chosen for the characterization of beta-galactosidase expression in bm5 and sf21 cells. expression level of beta-galactosidase and replicatio ... | 1997 | 9509418 |
| role of early and late replication events in induction of apoptosis by baculoviruses. | autographa californica nuclear polyhedrosis virus (acmnpv) mutants that lack the apoptotic suppressor gene p35 cause apoptosis in spodoptera frugiperda sf21 cells. to identify a viral signal(s) that induces programmed cell death, we first defined the timing of apoptotic events during infection. activation of a p35-inhibitable caspase, intracellular fragmentation of host and acmnpv dna, and cell membrane blebbing coincided with the initiation of viral dna synthesis between 9 and 12 h after infect ... | 1997 | 8995679 |
| expression of fas-fas ligand system associated with atresia in murine ovary. | we detected that fas receptor (fas) was expressed at transcriptional levels in oocytes/eggs and some granulosa cells of murine ovary, whereas, fas ligand (fasl) was found to be strongly expressed in granulosa cells by means of in situ hybridisation. these results were supported by an indirect immunofluorescence (iif) test with anti-fas monoclonal antibody (mab)/fasl ab. the lysates from granulosa cells were precipitated by anti-fasl ab, exhibiting a specific band at 40 kda. when zona pellucida ( ... | 1997 | 9563680 |
| expression, purification, and characterization of a soluble form of human cytomegalovirus glycoprotein b. | the human cytomegalovirus glycoprotein b gene (gb; gpul55) was truncated at amino acid 692 and recombined into autographa californica nuclear polyhedrosis virus (baculovirus). infection of insect cells with the recombinant baculovirus resulted in high-level expression and secretion of the truncated gb protein (gb-s) into the culture medium. purification of gb-s by monoclonal antibody affinity chromatography yielded a protein of ca. 200 kda. characterization of the 200-kda purification product in ... | 1997 | 9426459 |
| [expression of green fluorescent protein with baculovirus vector in insect cells]. | the green fluorescent protein (gfp) gene was subcloned into the transfer vector pvlneo downstream of the polyhedrin gene (ocu) promoter. insect cells were cotransfected with recombinant plasmid and autographa californica nuclear polyhedrosis virus (acnpv) dna. in the presence of g418, the recombinant virus containing gfp gene was purified. the gfp expressed in insect cells with a mw of 30 kda is observable by strong green light under a fluorescent microscope. excitation and emission spectra of t ... | 1997 | 9863195 |
| expression of cell surface gpi-anchored human p97 in baculovirus-infected insect cells. | the baculovirus/insect cell system (autographa californica multiple nuclear polyhedrosis virus/spodoptera frugiperda sf9 cells) was used to express the gpi-anchored human melanoma tumor antigen, melanotransferrin or p97. this system served to study the expression and productivity of recombinant gpi-anchored p97 by insect cells. the sf9 cells expressed a cell surface gpi-anchored form of p97 as well as a soluble form of p97 that did not appear to be derived from the gpi-anchored form of p97. both ... | 1997 | 18636443 |
| substrate limitation in the baculovirus expression vector system. | the inability to infect insect cell cultures at the highest achievable cell densities has imposed major limitations to both the fundamental understanding of the baculovirus expression vector system (bevs) as well as full exploitation of its potential productive capacity for recombinant (beta-galacnpv) products. the current literature does not characterize and identify the exact nature of the observed limitations, which therefore has become the major objective and contribution of the following st ... | 1997 | 18636607 |
| expression of green fluorescent protein in insect larvae and its application for heterologous protein production. | many eukaryotic proteins have been successfully expressed in insect cells infected with a recombinant baculovirus derived from the autographa californica nuclear polyhedrosis virus (acnpv). there are, however, disadvantages with this cell-based system when carried out in suspension cultures at high bioreactor volume (e.g., limited oxygen transfer, susceptibility to contamination, high cost). these problems can be avoided by using whole larvae as the "reactors." there are, however, other problems ... | 1997 | 18636639 |
| bcl-2 expression in spodoptera frugiperda sf-9 and trichoplusia ni bti-tn-5b1-4 insect cells: effect on recombinant protein expression and cell viability. | the effect of bcl-2 expression on cell viability and recombinant protein synthesis was investigated in the spodoptera frugiperda sf-9 and trichoplusia ni bti-tn-5b1-4 (high fivetrade mark) insect cell lines. it was found that coinfection with a baculovirus expressing bcl-2 [autographa californica nuclear polyhedrosis virus (acnpv)-bcl2] extended the life span of high fivetrade mark cells but not sf-9 cells when compared to infection with recombinant baculoviruses expressing either human tissue p ... | 1997 | 18642241 |
| nucleotide sequence analysis of hasnpv protein kinase. | the protein kinase gene of helicoverpa armigera single nucleocapsid nuclear polyhedrosis virus (hasnpv) has been cloned and sequenced. it is located approximately 1.25 kb downstream of the polyhedrin gene. the predicted molecular mass of this 267 amino acid protein (havpk) is 31 kd. havpk shows a 43.0% amino acid homology to vpk from lymantria dispar multinucleocapsid nuclear polyhedrosis virus (ldmnpv) and a 39.0% homology to pk-1 from autographa californica multinucleocapsid nuclear polyhedros ... | 1997 | 12219213 |
| studies on the expression of the 28 kd glutathione s-transferase gene from schistosoma japonicum in the silkworm (bombyx mori) larvae and insect cells. | the gene of the 28 kd glutathione s-transferase (gst) from the chinese strain of schistosoma japonicum had been expressed in the silkworm (bombyx mori) cells and larvae by bombyx mori nuclear polyhedrosis virus (bmnpv) vector which had been modified. the gst gene was inserted into the right position of the bmnpv genome as identified by southern hybridization. the product was a 28 kd protein, had gst activity and antigenicity. the yield in bmn cells (1x10(6) cells/ml) was 0.77 mg and more than 5 ... | 1997 | 12219234 |
| [biosynthesis of a single peptide chain containing human chorionic gonadotropin beta and ovine common alpha subunits tandem]. | hcg beta-olh alpha chimeric cdna was constructed by using overlapping pcr to contact the codons of c-terminal end of hcg beta with the codons of n-terminal end of olh alpha, then it was subcloned into nuclear polyhedrosis virus (acnpv) expression vector pvl1393 to construct expression vector pvl1393-hcg beta-olh alpha. the insect cells (sf9) were cotransfected by the expression vector pvl1393-hcg beta-olh alpha and baculogold acnpv linearized genomic dna, and recombinant viruses acnpv-hcg beta-o ... | 1998 | 12014113 |
| nucleotide sequence and characterization of the p10 gene of spodoptera litura nuclear polyhedrosis virus. | the p10 gene was localized within the 3.0 kbp bamh i fragment of the sinpv genome. with a coding sequence of 318 nucleotides, corresponding to a protein of 105 amino acids, the p10 gene of sinpv is the longest p10 gene identified so far in npvs. the sinpv p10 gene is also distinct in having two a/tttgta motifs in the promoter region. with 10 heptad repeats, the sinpv p10 protein is probably forming a relatively large coiled-coil structure. the organization of sinpv orf552-p10 gene-orf945 cluster ... | 1998 | 12167986 |
| expression of human lactoferrin cdna in insect cells. | a transfer plasmid p8hlfc harboring human lactoferrin cdna was constructed and co-transfected with autographa californica nuclear polyhedrosis virus ( acnpv bacpak6 ) dna into spodoptera frugiperda (sf) cells. the recombinant virus acnpv-hlfc was identified and purified three times by dot hybridization and plaques screening. the expression of human lactoferrin cdna in insect cells was detected by western blotting. the expression level of lactoferrin is about 2% of insect cellular total soluble p ... | 1998 | 12167991 |
| cloning of etp28 gene of eimeria tenella (guangdong strain) sporozoites and its expression in baculovirus expression system. | a refractile body antigen etp28 gene of eimeria tenella (guangdong strain) sporozoites was cloned by pcr from the synthesized first strand cdna. it has a high homology with the previously reported etp28 gene of merck strain ls18. the gene was expressed through standard procedures in the modified autographa californica nuclear polyhedrosis virus (acmnpv-occ(-)) expression system and large amount of heterologous fusion protein (gst-6xhis-etp28) was obtained. the expression product was about 21.3% ... | 1998 | 12174264 |
| 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 |
| 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 |
| 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 |
| 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 |
| generation of recombinant baculovirus dna in e.coli using a baculovirus shuttle vector. | baculovirus vectors are now widely used to direct the expression of heterologous genes in cultured insect cells and insect larvae. in most cases, heterologous genes placed under transcriptional control of the polyhedrm promoter of the autographa californica nuclear polyhedrosis virus (acnpv) are abundantly expressed during the late stages of infection. the recombinant proteins are usually soluble and functionally similar to their authentic counterparts (l-7). in the following sections, recent ad ... | 1998 | 21390847 |
| baculovirus-mediated gene transfer into mammalian cells. | a kind of cytotoxinic gene barnase was cloned downstream of immediate early promoter from cytomegalovirus (cmv). recombinant virus vaccmvbar was constructed using bac-to-bac system by insertion of the expression cassette into the polyhedrin gene locus of autographa californica nuclear polyhedrosis virus (acnpv). cytotoxinic effect caused by expression of barnase was observed in four mammalian cells after vaccmvbar infection dose-dependent analysis revealed that one of the cells hicam was the mos ... | 1998 | 18726228 |
| 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 |
| 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 |
| the p95 gene of bombyx mori nuclear polyhedrosis virus: temporal expression and functional properties. | as part of our effort to identify baculovirus proteins acting as transcriptional regulators, we have characterized a gene, p95, of bombyx mori nuclear polyhedrosis virus (bmnpv) that encompasses an open reading frame for a putative 95-kda polypeptide (p95). the n-terminal half of the conceptually translated p95 contains two zinc finger-type dna-binding motifs, and its c terminus contains a proline-rich region reminiscent of transcriptional activation regions. northern blot analysis indicates tha ... | 1998 | 9573244 |
| 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 |