| bovine respiratory syncytial virus nucleocapsid protein expressed in insect cells specifically interacts with the phosphoprotein and the m2 protein. | the coding region of the gene for the nucleocapsid (n) protein of bovine respiratory syncytial virus has been inserted into the genome of autographa californica nuclear polyhedrosis virus (acnpv) using transfer vector pvl 1393. infection of spodoptera frugiperda cells with recombinant virus resulted in the synthesis of high levels of n protein. this protein was indistinguishable from the authentic bovine respiratory syncytial virus (brsv) n protein by sds-gel electrophoresis and immunoprecipitat ... | 1993 | 8438581 |
| nucleotide sequence of the p74 gene of a baculovirus pathogenic to the spruce budworm, choristoneura fumiferana multicapsid nuclear polyhedrosis virus. | polypeptide p74 has been found to be essential for production of virulent occlusion bodies of the baculovirus autographa californica multicapsid nuclear polyhedrosis virus (acmnpv). hybridization with acmnpv-derived probes has led to the location of the p74 gene in the spruce budworm virus, choristoneura fumiferana mnpv. sequence data indicate that cfmnpv p74 is 73% identical to acmnpv at the nucleotide level and 77% identical at the amino acid level. elements of predicted secondary structure ar ... | 1993 | 8439559 |
| saturable attachment sites for polyhedron-derived baculovirus on insect cells and evidence for entry via direct membrane fusion. | this research provides the first evidence for specific receptor binding of polyhedron-derived baculovirus (pdv) to host cells and to lepidopteran brush border membrane vesicles (bbmv) and demonstration of entry via a nonendocytotic pathway involving direct membrane fusion. the technique of fluorescence-activated cell sorting analysis was used to investigate the specificity of binding between the pdv phenotype of lymantria dispar nuclear polyhedrosis virus (ldnpv) and host membranes. fluorescein ... | 1993 | 8445715 |
| three baculovirus genes involved in late and very late gene expression: ie-1, ie-n, and lef-2. | we developed a method to identify baculovirus genes required for late and very late gene expression that is based on subtraction of clones from an autographa californica nuclear polyhedrosis virus genomic library which is able to trans activate promoters of reporter plasmids in transient expression assays. using this assay, we found that three genes located between 83.7 and 7.5 map units of the autographa californica nuclear polyhedrosis virus genome were involved in expression from the late cap ... | 1993 | 8445724 |
| an apoptosis-inhibiting baculovirus gene with a zinc finger-like motif. | spodoptera frugiperda sf-21 cells infected with autographa californica nuclear polyhedrosis virus mutants which lack a functional p35 gene undergo apoptosis, a type of programmed cell death. to identify p35-homologous genes in other baculoviruses, a. californica nuclear polyhedrosis virus dna containing a deletion in p35 was cotransfected into sf-21 cells along with genomic dnas from other baculoviruses. one of the viral dnas which were able to rescue wild-type infection was from cydia pomonella ... | 1993 | 8445726 |
| nucleotide sequence and transcriptional analysis of a gene encoding gp41, a structural glycoprotein of the baculovirus autographa californica nuclear polyhedrosis virus. | | 1993 | 8445739 |
| ligand binding by a recombinant insect juvenile hormone binding protein. | a cdna for the hemolymph juvenile hormone binding protein (jhbp) of larval manduca sexta has been isolated, sequenced, and expressed in an insect cell line. a recombinant baculovirus, containing the jhbp cdna fused to the p10 promoter of autographa californica nuclear polyhedrosis virus, was constructed. insect cells (sf9) infected with this virus secreted recombinant jhbp (rjhbp) into the medium (> 50 micrograms/ml), and cotranslational removal of an 18 amino acid leader sequence was observed. ... | 1993 | 8448166 |
| functional domains of the p10 protein of autographa californica nuclear polyhedrosis virus. | distinct functional domains in the autographa californica nuclear polyhedrosis virus p10 protein were identified by analysis of p10 mutants. when up to 15 amino acids from the carboxy terminus were deleted, truncated p10 proteins were found in both the nucleus and the cytoplasm of infected cells, but formed no fibrillar structures. this suggested that the positively charged carboxy terminus is not required for nuclear or cytoplasmic localization of p10 protein, but is involved in protein-protein ... | 1993 | 8468550 |
| sequence, transcriptional mapping, and overexpression of p47, a baculovirus gene regulating late gene expression. | a 2.8-kb region of the autographa californica nuclear polyhedrosis virus genome was sequenced and found to contain an open reading frame (p47) which was capable of rescuing a previously characterized temperature-sensitive mutant, ts317 (s. partington, h. yu, a. lu, and e. b. carstens, virology 157:91-102, 1990). transcriptional mapping demonstrated that an early 4.2-kb rna encoded the p47 open reading frame and probably overlapped the 39k delayed-early gene. the p47 open reading frame was cloned ... | 1993 | 8474157 |
| the 35-kilodalton protein gene (p35) of autographa californica nuclear polyhedrosis virus and the neomycin resistance gene provide dominant selection of recombinant baculoviruses. | autographa californica nuclear polyhedrosis virus (acmnpv) recombinants were constructed to test the effectiveness of the acmnpv 35-kilodalton protein gene (35k gene) and the bacterial neomycin resistance gene (neo) as dominant selectable markers for baculoviruses. insertion of the acmnpv apoptosis suppressor gene (p35) into the genome of p35-deletion mutants inhibited premature host cell death and increased virus yields up to 1200-fold at low multiplicities in spodoptera frugiperda (sf21) cell ... | 1993 | 8493093 |
| identification and characterization of lef-1, a baculovirus gene involved in late and very late gene expression. | an autographa californica nuclear polyhedrosis virus (acmnpv) gene required in transient expression assays for late and very late viral gene expression was identified, sequenced, and transcriptionally mapped. this gene, designated late expression factor 1 (lef-1), was located between 7.4 and 8.7 map units of the acmnpv physical map. it was identified by cotransfecting spodoptera frugiperda cultured cells with a collection of overlapping cloned dna fragments covering the entire acmnpv genome and ... | 1993 | 8497062 |
| activation and detection of a latent baculovirus resembling mamestra brassicae nuclear polyhedrosis virus in m. brassicae insects. | a laboratory culture of mamestra brassicae insects (mblc) has been found to harbor a latent baculovirus infection. the latent virus was activated by feeding the m. brassicae larvae with either the closely related panolis flammea nuclear polyhedrosis virus (npv), or the distantly related autographa californica npv. restriction fragment profiles of the activated virus dna showed that it is very closely related, if not identical, to m. brassicae npv. polymerase chain reaction amplification of polyh ... | 1993 | 8503177 |
| efficient secretion of attacin from insect fat-body cells requires proper processing of the prosequence. | the attacins constitute a group of immune proteins induced after bacterial infection in the moth hyalophora cecropia. they are synthesized as preproproteins and undergo post-translational modification during transport to the hemolymph. the processing and transport rates of attacin were studied in its natural host as a response to infection. monensin totally inhibited the processing from proattacin to attacin and the radiolabeled proattacin remained intracellular. this observation indicated that ... | 1993 | 8508799 |
| nucleotide sequence and transcriptional analysis of the p10 gene of spodoptera exigua nuclear polyhedrosis virus. | the p10 gene of spodoptera exigua multiple nuclear polyhedrosis virus (semnpv) was localized on the xbai fragment h (5.1 kb) of the physical map of the viral genome. the coding sequence of the semnpv p10 gene is 264 nucleotides (nt) long corresponding to a predicted protein of 88 amino acids with an mhf of 9607. the semnpv p10 protein showed only limited amino acid identity (39% and 26%, respectively) to those of orgyia pseudotsugata mnpv (opmnpv) and autographa californica mnpv (acmnpv) and thu ... | 1993 | 8509757 |
| apoptosis reduces both the in vitro replication and the in vivo infectivity of a baculovirus. | apoptotic programmed cell death occurs when the insect cell line sf-21, derived from spodoptera frugiperda, is infected with mutants of the baculovirus autographa californica nuclear polyhedrosis virus (acmnpv) which lack a functional p35 gene. however, infection of the trichoplusia ni tn-368 cell line with p35 mutants does not result in apoptosis (r. clem, m. fechheimer, and l. miller, science 254:1388-1390, 1991). we have examined the effect of apoptosis on acmnpv infections in cell lines and ... | 1993 | 8510202 |
| in vitro transcription from baculovirus late gene promoters: accurate mrna initiation by nuclear extracts prepared from infected spodoptera frugiperda cells. | extracts prepared from nuclei of autographa californica nuclear polyhedrosis virus-infected spodoptera frugiperda cells were shown to support in vitro transcription from baculovirus late gene promoters. in vitro transcription was optimized for the late promoter of the 39k gene. the mg2+ concentration was critical; concentrations higher than 1 to 2 mm did not support late transcription. additional conditions included template (40 micrograms/ml), extract (2.5 mg/ml), and incubation time (25 min). ... | 1993 | 8510206 |
| a baculovirus-expressed fusion protein containing the antibody-binding domain of protein a and insect luciferase. | a fusion construct encoding two antibody-binding sites of protein a from staphylococcus aureus and click beetle, pyrophorus plagiophthalamus, luciferase (lucgr) was designed and expressed using the baculovirus system. the construct was inserted under the transcriptional regulation of the polyhedrin gene promoter of the autographa californica nuclear polyhedrosis virus (acnpv) and expressed in the insect spodoperta frugiperda cell line during viral infection. the properties of the resultant chime ... | 1993 | 8512706 |
| a method for producing recombinant baculovirus expression vectors at high frequency. | a system has been developed that can generate recombinant baculovirus expression vectors at frequencies approaching 100%. this system provides a selection for recombinant viruses by using the essential gene downstream of the autographa californica nuclear polyhedrosis virus (acmnpv) polyhedrin expression locus. two acmnpv derivatives were constructed in which the expression locus and part of the downstream gene are flanked by restriction sites. the parental viruses are viable; however, restricti ... | 1993 | 8512707 |
| a simple and efficient procedure for the oral inoculation of trichoplusia ni larvae with polyhedrin-negative recombinant baculovirus. | current procedures for inoculating lepidopteran larvae with polyhedrin-negative recombinant baculovirus, i.e. intracoelomic injection or coinfection with wild type virus, are laborious and can compromise final yields of recombinant protein. herein is described a simple and efficient method for oral inoculation. up to 100% infection was obtained when individual early fifth instar trichoplusia ni larvae were fed a small piece of a formaldehyde-free insect diet to which 4.2 x 10(5) pfu of a polyhed ... | 1993 | 8514841 |
| cloning and sequence analysis of a p40 structural protein gene of helicoverpa zea nuclear polyhedrosis virus. | a gene encoding an occluded virion structural protein was isolated from an expression library constructed from the helicoverpa zea s-type nuclear polyhedrosis virus (npv) isolate hzs-15 using both polyclonal and monoclonal antibody screening. the gene was located within a pstl-sall fragment of the hzs-15 genome spanning from 96.5 to 97.3 m.u. sequencing analyses revealed a long open reading frame of 927 nucleotides that predicted a protein of 37 kda in size. immunoblot analyses using the monoclo ... | 1993 | 8517018 |
| analysis of the role of 5' promoter elements and 3' flanking sequences on the expression of a baculovirus polyhedron envelope protein gene. | the polyhedron envelope protein gene of the orgyia pseudotsugata multinucleocapsid nuclear polyhedrosis virus (opmnpv) is the third in a series of five open reading frames (orfs 1-5) oriented in the same direction. individual mrnas initiate at conserved late gene promoter/mrna start site (a/gtaag) sequences located upstream of each orf and the mrnas co-terminate after the fifth orf. to examine the influence of transcription from upstream promoter elements and the presence of an extensive 3' flan ... | 1993 | 8517021 |
| a baculovirus encoded 16-kda glycoprotein localizes near the nuclear membrane of infected cells. | an open reading frame (orf 2) located upstream of the polyhedron envelope protein gene of orgyia pseudotsugata multicapsid nuclear polyhedrosis virus (opmnpv) was cloned in frame into a trpe expression vector. the fusion protein produced by this construct was used for the production of a monospecific antiserum. western blot analysis of opmnpv-infected lymantria dispar cells detected a 16-kda protein at 24 hr postinfection. the 16-kda protein was determined to be n-glycosylated by tunicamycin tre ... | 1993 | 8517029 |
| location of two putative origins of dna replication of autographa californica nuclear polyhedrosis virus. | previously, we described a defective autographa californica nuclear polyhedrosis virus (acmnpv) which must contain cis-acting elements required for dna synthesis, such as the origin(s) of replication (ori). defective genomes of acmnpv generated after serial undiluted passage were analyzed further. three small separated regions were retained in dna of defective acmnpv and accumulated in extracellular defective interfering viruses as well as in intracellular dna after 40 passages. two of these reg ... | 1993 | 8517035 |
| expression and radiolabeling of human c-reactive protein in baculovirus-infected cell lines and trichoplusia ni larvae. | human c-reactive protein (crp) is a member of the pentraxin family of proteins which are molecules composed of five identical subunits arranged in a planar configuration. in the present study a human crp cdna clone was ligated into the baculovirus vector pvl1393 which was used to establish a recombinant strain of baculogold autographa californica multiple nuclear polyhedrosis virus containing the coding and leader sequence for human crp (designated acmnpv-crp). synthesis and secretion of crp wer ... | 1995 | 8527929 |
| construction and insect larval expression of recombinant vesicular stomatitis nucleocapsid protein and its use in competitive elisa. | the gene encoding the nucleocapsid (n) protein of indiana 1 serotype vesicular stomatitis virus (vsv-in1) was transferred into the genome of autographa californica nuclear polyhedrosis virus (baculovirus) as a full-length non-fusion construct under the control of the polyhedrin gene promoter. recombinant n protein was obtained from trichoplusia ni insect larvae inoculated 72-96 h previously with the recombinant baculovirus. polyclonal antibody (pab) against vsv-in1 was produced in mice using vsv ... | 1995 | 8530566 |
| convenient desktop-scale production of the extracellular domain of the human growth hormone receptor by an insect-baculovirus secretion system using a protein-free culture. | expression of a gene encoding the extracellular domain of the human growth hormone receptor (hghr-ed) inserted into the genome of autographa californica nuclear polyhedrosis virus was done using a desktop-scale spinner culture. spodoptera frugiperda 9 (sf9) cells infected with the recombinant virus secreted a protein with hgh-binding activity into the medium. oxygen supplementation was required for high level secretion of the product. the highest cell production capability was estimated at more ... | 1995 | 8534995 |
| identification and characterization of the v-cath gene of the baculovirus, cfmnpv. | the v-cath gene of the autographa californica multi-nucleocapsid nuclear polyhedrosis virus (acmnpv) encodes a cathepsin l-like proteinase which plays a role in the liquefaction of host tissues during a viral infection [1]. we have identified a homologous gene in the spruce budworm virus, choristoneura fumiferana mnpv (cfmnpv). the cfmnpv v-cath gene is 74% identical to acmnpv v-cath at the nucleotide sequence level and 80% identical at the level of predicted amino acid sequence. transcription a ... | 1995 | 8547313 |
| high-level expression of human beta-interferon gene in the silkworm with new constructed bmnpv vector. | bombyx mori nuclear polyhedrosis virus (bmnpv) and bombyx mori cells as well as silkworm larvae were used successfully for the production of biologically active recombinant proteins. there are only a few types of bmnpv general vectors. here a new type of vector plasmid pbm92 was constructed in this experiment. the translational initiation codon atg of the polyhedrin gene in pbm92 was changed into att, and then five cloning sites of a foreign gene were ligated after the +12 bp site of the polyhed ... | 1995 | 8562846 |
| 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 ... | 1996 | 8567669 |
| production of polyhedra of the autographa californica nuclear polyhedrosis virus using the sf21 and tn5b1-4 cell lines and comparison with host-derived polyhedra by bioassay. | both wild-type and recombinant baculoviruses are becoming more attractive for the control of insect pests. thus, there is an increased incentive to address and resolve logistical problems associated with large-scale production of these viruses. in this study, we have compared the potential of two insect cell lines, tn5b1-4 and sf21, for the production of polyhedra and compared the efficacy of both cell culture-derived and host-derived viruses by bioassay. the efficacy of both wild-type acmnpv an ... | 1995 | 8568278 |
| nucleotide sequence of a 1446 base pair sali fragment and structure of a novel early gene of leucania separata nuclear polyhedrosis virus. | a 1446 bp sali fragment of lsnpv was sequenced by the silver staining method, and two large open reading frames (orfs, orf1 and orf2) were found, both contain typical characteristics of the 5' regulatory elements of baculovirus early genes. orf1 is 345 bp long with the capacity to encode a putative protein of 114 amino acid residues with mw about 13 kda and was designated p13 gene, orf2 comprises 248 bp from the 3' end of the fragment. in the untranslated region (utr) of orf1, a 33 bp mini cistr ... | 1995 | 8572949 |
| nucleocapsid protein n of lelystad virus: expression by recombinant baculovirus, immunological properties, and suitability for detection of serum antibodies. | the orf7 gene, encoding the nucleocapsid protein n of lelystad virus (lv), was inserted downstream of the p10 promoter into autographa californica nuclear polyhedrosis virus (baculovirus). the resulting recombinant baculovirus, designated bac-orf7, expressed a 15-kda protein in insect cells. this protein was similar in size to the n protein expressed by lv in cl2621 cells when it was analyzed on sodium dodecyl sulfate-polyacrylamide gels. the n protein expressed by bac-orf7 was immunoprecipitate ... | 1995 | 8574824 |
| two human alpha 2-adrenoceptor subtypes alpha 2a-c10 and alpha 2b-c2 expressed in sf9 cells couple to transduction pathway resulting in opposite effects on camp production. | the baculovirus expression vector system utilizing the strong polyhedrin gene promoter of the autographa californica nuclear polyhedrosis virus (acnpv) was used for high level expression of the two alpha 2-adrenoceptor subtypes alpha 2a-c10 and alpha 2b-c2 in spodoptera frugiperda (sf-9) insect cells. for rapid screening of recombinant viruses the luciferase gene was expressed under the early etl-promoter (early transcript large) in the same plasmid. both receptor subtypes showed the same rank o ... | 1995 | 8575536 |
| replication of a mamestra brassicae nuclear polyhedrosis virus in a newly established mamestra brassicae cell line. | a continuous cell line, designated mbl-3, was newly established from minced neonate larvae of the cabbage armyworm, mamestra brassicae. this new cell line is heteroploid, containing triploid cells predominantly at the frequency of about 50%. doubling time of the cell population is 33 hrs. the akutsu isolate of a mamestra brassicae nuclear polyhedrosis virus (mbnpv) was examined for replication in 16 continuous cell lines, including the cell line mbl-3, from seven lepidoptera species: mamestra br ... | 1995 | 8578995 |
| expression of human stem cell factor in the baculovirus expression system. | a partial cdna of human stem cell factor (scf) was isolated and transferred into the genome of bombyx mori nuclear polyhedrosis virus adjacent to the polyhedrin promoter. the soluble form of the scf protein was expressed in bombyx mori cells and silkworm larvae and was secreted into the medium in an active state. the recombinant scf was similar to natural scf with respect to its interaction with monoclonal antibody and its synergistic biological activity in conjunction with colony-stimulating fa ... | 1995 | 8589646 |
| the baculovirus cysteine protease has a cathepsin b-like s2-subsite specificity. | autographa californica nuclear polyhedrosis virus (acnpv) encodes a functional cysteine protease of the papain family which is expressed after infection in spodoptera fruglperda sf9 cells. the protease displays an inhibition profile typical for cysteine proteases and is highly active against synthetic peptide substrates. the ph optimum of the bell-shaped ph-activity curve is between 5.0 and 5.5. the best substrate tested is z-arg-arg-mca which is specific for cathepsin b. the specificity constan ... | 1995 | 8590630 |
| 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 ... | 1996 | 8599240 |
| 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 ... | 1996 | 8601794 |
| 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 ... | 1996 | 8607267 |
| 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 ... | 1996 | 8609477 |
| 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 ... | 1996 | 8609478 |
| 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 ... | 1996 | 8610448 |
| 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 |
| baculovirus-directed expression of human prostatic steroid 5 alpha-reductase 1 in an active form. | in the human prostate, the enzyme steroid 5 alpha-reductase (h5 alpha r) catalyses the conversion of testosterone into the more potent androgen, dihydrotestosterone. two distinct cdnas coding for h 5 alpha r in the human prostate have been previously characterized. enzyme h5 alpha r1 shows a maximum activity at basic ph whereas h5 alpha r2 has an acidic ph optimum activity. we report here the expression of the human steroid h5 alpha r1 in a eukaryotic expression system: the baculovirus-directed- ... | 1993 | 8664165 |
| 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 ... | 1996 | 8666274 |
| 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 ... | 1996 | 8676487 |
| high expression of hbv s gene in bombyx mori cell culture and in silkworms. | the coding sequence of human hepatitis b virus (hbv) subtype adr surface antigen s gene was inserted into the genome of the nuclear polyhedrosis virus of bombyx mori (bmnpv) under control of the polyhedrin promoter, and a recombinant bmnpvs virus was constructed. upon infection of the recombinant virus, the yield of hbsag was 35.5 micrograms per 10(6) cells, 750 micrograms per larva, and 690 micrograms per pupa. western blot and electron microscope observation of the preliminary purified hbsag p ... | 1995 | 8679930 |
| [sequencing of polyhedrin gene of leucania separata multiple capsid nuclear polyhedrosis virus by silver sequencing system]. | the genome of lsnpv was digested with ecorv and cloned into pbluescript. with the partial polyhedrin gene of autographa californica as a probe, positive recombinants were screened by colony in situ hybridization, further characterized by sequencing, and designated as plsev and plsph5. the intact 741 bp polyhedrin gene of lsnpv is located on the two recombinants revealed by silver sequencing system, and shares 80.0% and 97.0% nucleotide identity of the coding sequence, 87.8% and 97.5% amino acid ... | 1995 | 8701583 |
| 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 ... | 1996 | 8709251 |
| codon usage in the prototype baculovirus--autographa californica nuclear polyhedrosis virus. | the overall codon usage profile of autographa californica nuclear polyhedrosis virus (acnpv) was analyzed, using uwgcg sequence analysis software package from the known protein coding gene sequences available in genbank release 72. the analysis revealed that although only 45% of the codon used by acnpv have g/c at wobble base position, 15 out of 20 acnpv codons over-utilized for their given amino acids has g/c at the wobble position indicating a possible selection of these codons. the difference ... | 1995 | 8714213 |
| 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 ... | 1996 | 8738171 |
| [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. | 1996 | 8743017 |
| 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 ... | 1996 | 8760466 |
| 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 ... | 1996 | 8764020 |
| 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 ... | 1996 | 8776764 |
| 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 ... | 1996 | 8793839 |
| 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 ... | 1996 | 8794314 |
| 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 ... | 1996 | 8794340 |
| 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 ... | 1996 | 8794358 |
| 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. | 1996 | 8799322 |
| 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 ... | 1996 | 8806491 |
| 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 ... | 1996 | 8806500 |
| 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 ... | 1996 | 8806516 |
| 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 ... | 1996 | 8806578 |
| identification and nucleotide sequence of an ecdysteroid udp-glucosyltransferase gene of spodoptera littoralis multicapsid nuclear polyhedrosis virus. | the spodoptera littoralis multicapsid nuclear polyhedrosis virus (slmnpv) is a member of the baculoviridae that shows a distant genetic relationship to the prototype autographa californica mnpv (acmnpv). using an acmnpv gene-specific probe, we identified and mapped an ecdysteroid udp-glucosyltransferase (egt) gene in the genome of slmnpv. sequence determination of a part from the hybridizing dna fragment revealed an open reading frame of 1548 nucleotides that exhibits 38% and 44% identity to the ... | 1995 | 8808334 |
| 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. ... | 1996 | 8811031 |
| continuous cell line from pupal ovary of perina nuda (lepidoptera: lymantriidae) that is permissive to nuclear polyhedrosis virus from p. nuda | a 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 ... | 1996 | 8812599 |
| 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 ... | 1996 | 8812862 |
| in vivo and in vitro characterization of several isolates of spodoptera exigua nuclear polyhedrosis virus. | spodoptera exigua nuclear polyhedrosis viruses (senpvs), isolated form five geographically distinct regions of japan and thailand, were characterized by their dna restriction endonuclease pattern, level of virus production in a continuous cell line of s. exigua and biological activity to s. exigua larvae. the ecori and psti fragments exhibited similar overall patterns with minor differences. digestion of virus dna from a plaque-purified isolate, senpv-i1, with psti yielded 14 fragments and the e ... | 1995 | 8825303 |
| establishment of a cell line derived from embryos of the potato tuber moth phthorimaea operculella (zeller). | a cell line from the main insect pest of potatoes in tropical and subtropical areas, phthorimaea operculella (zeller), was obtained from embryoculture. these cells were cultured in grace's modified medium. the cell line, designated ors-pop-93, had a heterogeneous population consisting of spherical and spindle cells with great capacity to adhere and a doubling time of 40 h. they were subcultured for more than 60 passages. their polypeptidic profile was different from profiles of other lepidoptera ... | 1995 | 8826086 |
| 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 ... | 1996 | 8831686 |
| 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 ... | 1996 | 8836032 |
| characterization of the baculovirus choristoneura fumiferana multicapsid nuclear polyhedrosis virus p10 gene indicates that the polypeptide contains a coiled-coil domain. | the dna sequence and transcription pattern of the p10 gene from the spruce budworm baculovirus choristoneura fumiferana multicapsid nuclear polyhedrosis virus (cfmnpv) were analysed. the cfmnpv p10 gene codes for a protein 81 amino acids in length: this is the shortest p10 peptide identified thus far. a novel characteristic of the cfmnpv p10 gene is that it contains tandem late initiation sites (taag) having different temporal transcription patterns. comparative analysis of cfmnpv p10 and the am ... | 1995 | 8847496 |
| identification and characterization of an early gene in the lymantria dispar multinucleocapsid nuclear polyhedrosis virus. | the lymantria dispar multinucleocapsid nuclear polyhedrosis virus (ldmnpv) gene encoding g22 was cloned and sequenced. the g22 gene codes for a 191 amino acid protein with a predicted m(r) of 22,000. expression of g22 in a rabbit reticulocyte system generated a protein with an m(r) of 24,000, in close agreement with the molecular mass predicted from the nucleotide sequence. g22 is not significantly homologous to any known protein, nor is a g22 homologue present in the autographa californica mnpv ... | 1995 | 8847497 |
| 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 ... | 1996 | 8860651 |
| 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 ... | 1996 | 8862393 |
| 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 ... | 1996 | 8862418 |
| 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- ... | 1996 | 8910443 |
| 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 ... | 1996 | 8910650 |
| 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 ... | 1996 | 8914851 |
| 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 ... | 1996 | 8918923 |
| 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. | 1996 | 8921880 |
| 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 ... | 1996 | 8930653 |
| 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 ... | 1996 | 8941320 |
| 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 ... | 1996 | 8943013 |
| 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 ... | 1996 | 8950470 |
| 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 ... | 1996 | 8969454 |
| 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 |