Publications
| Title | Abstract | Year(sorted ascending) Filter | PMID Filter |
|---|
| expression of the gene encoding firefly luciferase in insect cells using a baculovirus vector. | a cdna encoding the firefly luciferase [photinus luciferin: oxygen 4-oxidoreductase (decarboxylating, atp-hydrolyzing), ec 1.13.12.7] was cloned downstream from the polyhedrin gene promoter of autographa californica nuclear polyhedrosis virus and expressed in spodoptera frugiperda clone-9 cells. synthesis of luciferase (luc) was accurately measured in insect cells growing in a 96-well plate, by a simple, rapid, nonradioactive, inexpensive and sensitive method based on fogging of x-ray film. luc ... | 1990 | 2205537 |
| differential gene expression mediated by late, very late and hybrid baculovirus promoters. | a series of recombinant viruses was constructed to study the regulation of expression from the promoter (pcap) for the major capsid-protein-encoding gene (vp39) of the baculovirus, autographa californica nuclear polyhedrosis virus. each virus of this series contains the cat reporter gene under the control of pcap or portions thereof. pcap regulation of cat was determined at the rna and enzyme activity levels, and was compared to cat gene expression driven by ppolh, the promoter of the polyhedrin ... | 1990 | 2205543 |
| the expression of the autographa californica nuclear polyhedrosis virus genome in insect cells. | this report presents a synopsis of recently published work in our laboratory on the molecular biology of the insect baculovirus autographa californica nuclear polyhedrosis virus (acnpv). the following studies have been summarized. (1) on the mode of transcription of the acnpv genome in insect cells. (2) translation of proteins encoded in the 81.2 to 85.0 map unit segment of acnpv. (3) inserts of insect cell dna in the acnpv genome. (4) expression of influenza (fowl plague) virus haemagglutinin i ... | 1990 | 2205973 |
| expression and characterization of the chick nicotinic acetylcholine receptor alpha-subunit in insect cells using a baculovirus vector. | a baculovirus transfer vector was constructed containing an entire cdna copy of the chick nicotinic acetylcholine receptor (nachr) alpha-subunit under control of the autographa californica nuclear polyhedrosis virus (acnpv) polyhedrin gene promoter. recombinant baculovirus was obtained by co-transfection of spodoptera frugiperda cells with infectious, wild-type acnpv dna and the transfer vector. polyhedrin-negative, recombinant viruses were identified which expressed the nachr alpha-subunit. the ... | 1990 | 2209600 |
| linearization of baculovirus dna enhances the recovery of recombinant virus expression vectors. | engineered derivatives of autographa californica multiple nucleocapsid nuclear polyhedrosis virus (acmnpv) possessing a unique restriction site provide a source of viral dna that can be linearized by digestion with a specific endonuclease. circular or linearized dna from two such viruses were compared in terms of their infectivity and recombinogenic activities. the linear forms were 15- to 150-fold less infectious than the corresponding circular forms, when transfected into spodoptera frugiperda ... | 1990 | 2216760 |
| expression, characterization, and purification of a phosphorylated rabies nucleoprotein synthesized in insect cells by baculovirus vectors. | a baculovirus expression vector (acnpv3) derived from the nuclear polyhedrosis virus of autographa californica (acnpv) was prepared containing the complete coding region of the nucleoprotein (n) gene of rabies virus (gif-sur-yvette clone of the cvs strain). the gene was placed under the control of the acnpv polyhedrin promoter and was expressed to high levels (66 mg n protein/liter of 2 x 10(9) cells) by the derived recombinant virus using a spodoptera frugiperda cell line. using available antis ... | 1990 | 2219704 |
| nucleotide sequence and characterization of the 39k gene region of autographa californica nuclear polyhedrosis virus. | the complete nucleotide sequence of the pstl-k fragment of the autographa californica nuclear polyhedrosis virus (acmnpv) genome was determined. this region of the genome contains the delayed early 39k gene and v-ubi, a viral protein with homology to ubiquitine. in addition, pstl-k potentially encodes five other proteins. hybrid-select translation mapped nine viral-specific proteins to this fragment. three proteins were observed with both early and late rna; two of them comigrated with proteins ... | 1990 | 2219715 |
| expression of cauliflower mosaic virus gene i using a baculovirus vector based upon the p10 gene and a novel selection method. | a new baculovirus expression vector based upon the p10 gene of autographa californica nuclear polyhedrosis virus (acnpv) and a novel system for the screening of p10 recombinants have been developed. the insertion of a cassette containing the lacz gene under the control of a heat-shock promoter of drosophila melanogaster downstream from the cloning site in p10 transfer vectors allows the convenient identification of putative recombinants by virtue of their expression of beta-galactosidase. using ... | 1990 | 2219726 |
| expression of cauliflower mosaic virus gene i in insect cells using a novel polyhedrin-based baculovirus expression vector. | an improved polyhedrin-based baculovirus expression vector was constructed to expedite distinguishing infections by putative baculovirus recombinants from infections by wild-type (wt) baculovirus. the vector utilizes the escherichia coli beta-galactosidase gene (lacz) as a genetic marker for positive recombination between wt autographa californica nuclear polyhedrosis virus and the baculovirus transfer vector. the marker gene/expression cassette was constructed so that lacz and the deleted polyh ... | 1990 | 2230725 |
| [purification of autographa californica nuclear polyhedrosis virus dna and cloning of the polyhedrin gene]. | autographa califonica nuclear polyhedrosis virus (acnpv) dna from larvae of lepidopteran infected by acnpv has been isolated. the ecori fragments of acnpv dna have been cloned using plasmid pbr325 as a vector. the polyhedrin gene fragment containing recombinant plasmid pac-18 was isolated through in situ hybridization. for the convenience of further construction the ecori insertion fragment was cloned into puc-8. thus plasmid puac-2 was obtained. the results of restriction mapping of pac-18 and ... | 1990 | 2242284 |
| identification and characterization of a baculovirus occlusion body glycoprotein which resembles spheroidin, an entomopoxvirus protein. | a 37-kda polypeptide specified by autographa californica nuclear polyhedrosis virus was found to share significant homology with choristoneura biennis entomopoxyvirus spheroidin protein, which is the major component of entomopoxvirus occlusion bodies. antibodies raised against spheroidin cross-reacted with the 37-kda protein and confirmed its expression in the late phase of wild-type baculovirus infection. immunoblot analysis and fluorescence microscopy demonstrated that the protein was associat ... | 1990 | 2243377 |
| infected cell specific protein and viral dna synthesis in productive and abortive infections of spodoptera frugiperda nuclear polyhedrosis virus. | this study examines viral protein and dna synthesis in spodoptera frugiperda multicapsid nuclear polyhedrosis virus (sfmnpv) infections of s. frugiperda and trichoplusia ni cells. a total of 28 infected cell specific polypeptides (icsps) were detected in the productive s. frugiperda cells. of these, 14 were identified as structural polypeptides. based on the change in their rate of synthesis during the replication cycle, these icsps were grouped into four classes. only a 97k and a 29k icsp were ... | 1990 | 2248550 |
| expression of plant virus genes in animal cells: high-level synthesis of cowpea mosaic virus b-rna-encoded proteins with baculovirus expression vectors. | the baculovirus expression system has been used to produce non-structural proteins encoded by bottom-component rna (b-rna) of cowpea mosaic virus (cpmv). for this, cdnas containing the 60k, 87k, 110k and 170k protein coding sequences were each provided with an atg start codon and the cdna containing the 60k coding sequence with a taa stop codon immediately downstream of the coding sequence. recombinant baculoviruses were retrieved which harboured the modified b-cdna sequences under the control o ... | 1990 | 2254749 |
| characteristically distinct isolates of the nuclear polyhedrosis virus from spodoptera litura. | more than 100 isolates were plaque-purified to examine the genetic variations in four wild stocks of spodoptera litura nuclear polyhedrosis virus (npv) collected in japan. these isolates were characterized by their in vitro host range in three established insect cell lines, growth characteristics, polyhedral protein, dna restriction endonuclease pattern and dna hybridization. the isolates were separated into four distinct groups: (i) isolates corresponding to autographa californica npv, (ii and ... | 1990 | 2254753 |
| characterization of the infection cycle of the orgyia pseudotsugata multicapsid nuclear polyhedrosis virus in lymantria dispar cells. | to characterize the infection cycle of the orgyia pseudotsugata multicapsid nuclear polyhedrosis virus in lymantria dispar cells, the time course of dna synthesis and polyhedron production, and the onset and rate of budded virus production were investigated at three different m.o.i. (5, 10 and 100). in addition, the time course of expression of three proteins (gp64, p39 and polyhedrin) representative of three temporal classes of baculovirus genes was also analysed using western blot analysis. dn ... | 1990 | 2273385 |
| nucleotide sequence and transcriptional analysis of the hindiii p region of a temperature-sensitive mutant of autographa californica nuclear polyhedrosis virus. | dna sequence analysis of the hindiii p region of a temperature-sensitive mutant of autographa californica nuclear polyhedrosis virus confirmed the specific amplification of 1.4 kb of viral dna from this region of the genome. the sequenced region included an open reading frame, translated in a counterclockwise direction, which would potentially encode a 74k protein. the amplified dna was contained within this open reading frame, resulting in in-frame amplifications of a domain within the protein. ... | 1990 | 2273394 |
| nucleotide sequence of a circular single-stranded dna associated with coconut foliar decay virus. | a circular single-stranded (ss) covalently closed (ccc) dna associated with coconut foliar decay virus (cfdv) was purified, amplified by the polymerase chain reaction, and subcloned and its sequence established by analysis of overlapping subgenomic cdna clones. the complete cfdv sequence comprised 1291 nucleotides and contained open reading frames for six proteins of molecular weight larger than 5 kda. one of these (orf1, 33.4 kda) codes for a leucine-rich protein with the nucleoside triphosphat ... | 1990 | 2345968 |
| expression of human granulocyte-macrophage colony-stimulating factor gene in insect cells by a baculovirus vector. | a plasmid pac373gm-csf was constructed and co-transfected into spodoptera frugiperda (sf9) cells with wild-type autographa californica nuclear polyhedrosis virus (acnpv) dna. the recombinant virus vac373gm-csf was identified and purified by several rounds of plaque hybridization. by assaying the culture medium, we demonstrated recombinant virus infected sf9 cells expressing hgm-csf. recombinant hgm-csfs with apparent molecular masses of 14.5, 15.5 and 16.5 kda were detected by the western blot m ... | 1990 | 2403525 |
| a baculovirus polyhedron envelope protein: immunogold localization in infected cells and mature polyhedra. | a polyclonal antiserum against a trpe fusion protein containing the complete open reading frame of the polyhedron envelope (pe) protein from the nuclear polyhedrosis virus of orgyia pseudotsugata (opmnpv) was used for immunogold staining and electron microscopic examination of polyhedra, isolated polyhedron envelopes, and infected insect cells at selected times postinfection. the antiserum specifically stained the peripheral envelope of mature polyhedra and also stained the envelope structure wh ... | 1990 | 2403704 |
| identification of virus-specific polypeptides and translatable mrnas in the isolated pupal abdomens of the silkworm, bombyx mori, infected with nuclear polyhedrosis virus. | analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis demonstrated that infection of the isolated pupal abdomens of the silkworm, bombyx mori, with b. mori nuclear polyhedrosis virus (bmnpv) caused generation of a number of polypeptides with a concomitant decrease of cellular polypeptides. these generated polypeptides were not identified as viral structural polypeptides, but were characterized as the degradation products of cellular polypeptides, as evidenced by the reaction with ... | 1990 | 2405063 |
| rate of increase of autographa californica nuclear polyhedrosis virus in trichoplusia ni larvae determined by dna:dna hybridization. | the rate of increase and doubling time of the hob clone of autographa californica nuclear polyhedrosis virus (acmnpv-hob) in neonate trichoplusia ni larvae was determined by measuring the increase in viral dna through time following inoculation with average doses of 50 or 17,400 occlusion bodies per larva. changes in total dna and viral dna through time were followed by fluorescence spectroscopy and quantitative slot-blot dna:dna hybridization, respectively. total dna content (i.e., larval dna a ... | 1990 | 2405064 |
| structure of preproattacin and its processing in insect cells infected with a recombinant baculovirus. | from a cdna library made from fatbody of immunized hyalophora cecropia pupae, a full-length clone corresponding to basic attacin was isolated. the corresponding amino acid sequence suggests that basic attacin is synthesized as a 233-residue preproprotein. this was confirmed by cloning the cdna fragment encoding the basic attacin in the baculovirus autographa californica nuclear polyhedrosis virus downstream of the polyhedrin promoter and expressing the protein in spodoptera frugiperda (sf9) cell ... | 1990 | 2406140 |
| expression of the lassa virus nucleocapsid protein in insect cells infected with a recombinant baculovirus: application to diagnostic assays for lassa virus infection. | the coding region of the gene for the nucleocapsid protein of lassa virus has been inserted into the genome of autographa californica nuclear polyhedrosis virus (acnpv) using the transfer vector pacym1, so that expression of the foreign dna is under the control of the promoter of the acnpv polyhedrin gene. infection of cultured spodoptera frugiperda cells with recombinant virus resulted in the synthesis of high levels of a protein that was indistinguishable from the authentic lassa virus protein ... | 1990 | 2406367 |
| overlapping divergent transcripts mapping to the hindiii f region of the autographa californica nuclear polyhedrosis virus. | we have mapped a set of transcripts that cross the xhoi site in the hindiii f fragment of the genome of the autographa californica nuclear polyhedrosis virus. these transcripts overlap at their 5' ends by about 550 bases and run in opposite directions. we have tentatively identified two open reading frames corresponding to the leftward transcript. the rightward transcript is present from 8 h post-infection (p.i.) to 24 h p.i.; the leftward transcripts are present from 2 h p.i. to 24 h p.i. the e ... | 1990 | 2406368 |
| functional analysis of a 603 nucleotide open reading frame upstream of the polyhedrin gene of autographa californica nuclear polyhedrosis virus. | the sequence of the 2000 nucleotides immediately upstream of the polyhedrin gene of the autographa californica multiple nucleocapsid nuclear polyhedrosis virus has been determined. comparative analysis of the data identified a 603 nucleotide open reading frame (orf) separated from the polyhedrin gene coding sequences by 156 nucleotides and in the opposite strand of dna. northern hybridization analysis of polyadenylated rna from infected cells highlighted a 3.7 kb species produced maximally at 12 ... | 1990 | 2407804 |
| autographa californica m nuclear polyhedrosis virus: microtubules and replication. | progressive reorganization and depolymerization of microtubules corresponded with virus-induced rounding of autographa californica m nuclear polyhedrosis virus (acmnpv)-infected spodoptera frugiperda iplb-sf-21 cells, suggesting that microtubules were instrumental in maintaining the normal shape of these cells. depolymerization of all cortical and most of the paranuclear microtubules with colchicine also resulted in cell rounding, confirming this hypothesis. studies with aphidicolin and cyclohex ... | 1990 | 2408230 |
| isolation of temperature sensitive mutants of autographa californica nuclear polyhedrosis virus: phenotype characterization of baculovirus mutants defective in very late gene expression. | temperature sensitive mutants of autographa californica nuclear polyhedrosis virus were isolated following exposure of parental replicating virus to 5-bromo-2'-deoxyuridine. two mutants designated ts317 and ts538 were characterized in detail. both mutants synthesized viral dna but were defective in infectious extracellular virus production at the nonpermissive temperature suggesting defects in late gene regulation. pulse-labeling experiments indicated that a switch to late protein synthesis occu ... | 1990 | 2408231 |
| transcription of the baculovirus polyhedrin gene reduces the levels of an antisense transcript initiated downstream. | a late 3.2-kilobase (kb) rna initiated approximately 2 kb downstream of the 3' end of the autographa californica nuclear polyhedrosis virus polyhedrin-coding sequence and traversed the polyhedrin gene in an antisense direction. this rna was sense rna for the two open reading frames flanking the polyhedrin gene. a mutant virus, vxpoly, which differs from wild-type virus only at the essential rna initiation site in the polyhedrin promoter, exhibited higher levels of the 3.2-kb rna than did wild-ty ... | 1990 | 1692351 |
| gene organization and transcription of ted, a lepidopteran retrotransposon integrated within the baculovirus genome. | a single copy of the retrotransposon ted, from the moth trichoplusia ni (a lepidopteran noctuid), was identified within the dna genome of the baculovirus autographa californica nuclear polyhedrosis virus. determination of the complete nucleotide sequence (7,510 base pairs) of the integrated copy indicated that ted belongs to the family of retrotransposons that includes drosophila melanogaster elements 17.6 and gypsy and thus represents the first nondipteran member of this invertebrate group to b ... | 1990 | 1692964 |
| categorizing some early and late transcripts directed by the autographa californica nuclear polyhedrosis virus. | using an s1 mapping assay on rna from spodoptera frugiperda cells infected by the autographa californica nuclear polyhedrosis virus in the presence and absence of cycloheximide and aphidicolin, we can distinguish three classes of transcripts. first, there are those whose synthesis is blocked by the dna synthesis inhibitor aphidicolin and which are therefore late transcripts. these include the late transcript of the 39k gene and a late leftward transcript across the xhoi site in the hindiii-f reg ... | 1990 | 1698929 |
| characterization of virus-like particles produced by a recombinant baculovirus containing the gag gene of the bovine immunodeficiency-like virus. | the entire gag gene of the bovine immunodeficiency-like virus (biv) was inserted behind the strong polyhedron promoter of autographa californica nuclear polyhedrosis virus (acnpv). the resultant recombinant baculovirus (acnpv-bivgag) was used to infect insect cells in order to overexpress and characterize biv gag gene products. the infection resulted in the high-level expression of a protein similar in size to the predicted biv gag precursor (pr53gag). biv pr53gag was detected in acnpv-bivgag-in ... | 1990 | 1699349 |
| continuous production of baculovirus in a cascade of insect-cell reactors. | insect cells (spodoptera frugiperda) were cultured in a continuous stirred-tank reactor. the effluent was led to a cascade of another two reactors, each containing half the volume of the cell-growth reactor, where the cells were infected with autographa californica nuclear polyhedrosis virus. for about 10 days production of 10(7) polyhedra (virus particles embedded in a protein capsule) per cm3 was achieved. this short production time compared to previous experiments involving an analogous syste ... | 1990 | 1366563 |
| multistage production of autographa californica nuclear polyhedrosis virus in insect cell cultures. | the aim of our study was to establish an efficient system for the in vitro production of the insect pathogenic autographa californica nuclear polyhedrosis virus in a spodoptera frugiperda cell line. we optimized cultivation conditions for cell proliferation as well as for virus replication in a 1.5 litre stirred tank bioreactor. cell and virus propagation were found to be optimal at a constant oxygen tension of 40%. in order to provide sufficient nutrients during virus synthesis filtration and p ... | 1990 | 1366992 |
| dna distribution and respiratory activity of spodoptera frugiperda populations infected with wild-type and recombinant autographa californica nuclear polyhedrosis virus. | spodoptera frugiperda cells were infected with a wild-type autographa californica nuclear polyhedrosis virus and with a recombinant autographa californica nuclear polyhedrosis virus. the recombinant virus was derived from the wild-type virus and produced beta-galactosidase instead of polyhedrin. the changes in cell size, cell growth, viability, dna distribution, and respiratory activity were followed through the time course of the infection. the dna content as measured by flow cytometry of infec ... | 1990 | 1367465 |
| simulated acid rain reduces the susceptibility of the european pine sawfly (neodiprion sertifer) to its nuclear polyhedrosis virus. | the study dealt with the effect of simulated acid rain (both h(2)so(4) and hno(3); acidities of ph 4 and ph 3) on the susceptibility of the larvae of neodiprion sertifer to its nuclear polyhedrosis virus. scots pines growing in a subarctic area with low ambient pollution levels were irrigated with simulated acid rain during two summers. neodiprion larvae fed with foliage from the experimental trees were infected with a dilute virus suspension. the acid treatment of host trees had a significant e ... | 1990 | 22160113 |
| purification and functional characterization of the human beta 2-adrenergic receptor produced in baculovirus-infected insect cells. | a human cdna fragment bearing the complete coding region for the beta 2-adrenergic receptor was introduced into the genome of autographa california nuclear polyhedrosis virus under the control of the polyhedrin promoter. binding studies using [125i]iodocyanopindolol showed that sf9 insect cells infected with the recombinant virus expressed approximately 1 x 10(6) beta 2-adrenergic receptors on their cell surface. photoaffinity labeling of whole cells and membranes revealed a molecular weight of ... | 1991 | 1645288 |
| secretion of biologically active leech hirudin from baculovirus-infected insect cells. | the thrombin inhibitor, hirudin, from the leech hirudo medicinalis, is the most powerful natural anticoagulant known. it has been characterized as a polypeptide of 65 amino acids which exhibits its anticoagulant properties by binding tightly and specifically to alpha-thrombin. the potency and specificity of hirudin have generated interest on its possible use in the treatment or prophylaxis of various thrombotic diseases. we have used the baculovirus expression system to efficiently produce activ ... | 1991 | 1647362 |
| high level expression of dna polymerases from herpesviruses. | the dna polymerase genes of human cytomegalovirus (hcmv) and varicella-zoster virus (vzv) were inserted separately into the polyhedrin gene of autographa californica nuclear polyhedrosis virus (acnpv) by cotransfection of spodoptera frugiperda (sf9) cells with baculovirus transfer vectors carrying the genes and acnpv infectious dna. infection of sf9 cells with the recombinant viruses resulted in expression from the polyhedrin promoter of proteins of the expected mrs. these proteins possessed dna ... | 1991 | 1649906 |
| insect virus: assays for viral replication and persistence in mammalian cells. | viral pesticidal agents must be evaluated for potential health hazards prior to utilization. assessment of the likelihood of replication in humans has included in vitro exposure of human cells to the potential pesticidal agent. previous in vitro evaluation strategies have lacked positive controls. thus, negative results, interpreted as no effect of the virus on human cells, could reflect basic deficiencies in the testing protocols. we designed a testing scheme for viral pesticides and used it to ... | 1991 | 1650786 |
| functional expression of the human growth factor activatable na+/h+ antiporter (nhe-1) in baculovirus-infected cells. | we constructed a recombinant baculovirus, based on autographa californica nuclear polyhedrosis virus, containing the human na+/h+ antiporter cdna under control of the polyhedrin promoter. when infected with this recombinant baculovirus, the sf9 cell line, derived from spodoptera frugiperda, expresses a fully functional na+/h+ antiporter as measured by the generation of an amiloride-sensitive na+ influx in response to an acid load. the na+/h(+)-exchange activity, not detectable in sf9 cells, emer ... | 1991 | 1654101 |
| processing and antigenicity of entire and anchor-free spike glycoprotein s of coronavirus tgev expressed by recombinant baculovirus. | the s gene of transmissible gastroenteritis virus (tgev) was inserted into the genome of autographa californica nuclear polyhedrosis virus (acnpv) using the transfer plasmid pvl941. infection of sf9 insect cells with the recombinant virus resulted in the synthesis of a 175k polypeptide which was able to trimerize and was transported to the cell surface as is the authentic tgev s protein. despite the lack of complete carbohydrate processing, the recombinant s protein exhibited antigenic propertie ... | 1991 | 1660201 |
| a functional hepatitis b virus x protein produced in insect cells. | we have constructed a recombinant baculovirus from autographa californica nuclear polyhedrosis virus, called acx, that expresses the gene encoding the hepatitis b virus x protein in infected spodoptera frugiperda (sf21ae) insect cells. a 16.5-kda monomer and a 33-kda dimer of the x protein were detected in extracts from acx-infected cells on immunoblots using a polyclonal anti-x antibody. the biological activity of the insect cell-produced x protein was assayed by fusing acx-infected sf21ae cell ... | 1991 | 1660211 |
| characterization of pseudorabies virus neutralization antigen glycoprotein giii produced in insect cells by a baculovirus expression vector. | the gene encoding the complete glycoprotein of pseudorabies virus (prv, yamagata-s81 strain glycoprotein giii) has been inserted into the baculovirus transfer vector pacym1s derived from the nuclear polyhedrosis virus of autographa californica (acnpv). a spodoptera frugiperda cell line, sf21ae, was efficiently co-transfected with the transfer vector containing the giii gene and acnpv dna by cationic liposomes (lipofectin). the gene was placed under the control of the acnpv polyhedrin promoter an ... | 1991 | 1661981 |
| immunoselection of recombinant baculoviruses expressing high levels of biologically active herpes simplex virus type 1 glycoprotein d. | the dna sequence encoding the complete herpes simplex virus type 1 (hsv-1) glycoprotein d (gd) was inserted into a baculovirus transfer vector under control of the polyhedrin gene promoter of the baculovirus autographa california nuclear polyhedrosis virus (acnpv). after co-transfection of spodoptera frugiperda (sf9) insect cells with wild-type acnpv dna and the recombinant transfer vector dna, polyhedrin-negative recombinants that expressed high levels of hsv-1 gd were isolated using immunoaffi ... | 1991 | 1662037 |
| dna restriction polymorphism in wild isolates of spodoptera frugiperda nuclear polyhedrosis virus. | restriction endonuclease analysis was used to examine variation in dna of 22 wild isolates of spodoptera frugiperda nuclear polyhedrosis virus (sfnpv). eleven of the 15 isolated from louisiana were distinguishable based on restriction fragment profiles from the enzymes bamhi, hindiii, and ecori. there was significant genetic variation in sfnpv isolates within single agricultural fields. nucleotide sequence divergence values, based on restriction fragment profiles, indicated that genetic variatio ... | 1991 | 1679452 |
| temporal analysis and spatial mapping of lymantria dispar nuclear polyhedrosis virus transcripts and in vitro translation polypeptides. | genomic expression of the lymantria dispar multinucleocapsid nuclear polyhedrosis virus (ldmnpv) was studied. viral specific transcripts expressed in cell culture at various times from 2 through 72 h postinfection were identified and their genomic origins mapped through northern analysis. sixty-five distinct transcripts were identified in this analysis. most viral transcripts were expressed late in infection, and originated from throughout the viral genome. viral polypeptides expressed in infect ... | 1991 | 1685048 |
| the basic dna-binding protein of bombyx mori nuclear polyhedrosis virus: the existence of an additional arginine repeat. | the basic dna-binding protein of the bombyx mori nuclear polyhedrosis virus (bmnpv) was purified by hplc and a sequence of 45 amino acids from the n-terminus was determined. there were no detectable modifications such as n-terminal blockage, glycosylation, or phosphorylation. the amino acid sequence showed high homology to the predicted amino acid sequences of the basic proteins of autographa californica npv (acnpv) and orgyia pseudotsugata npv (opnpv) (90 and 76%, respectively), however, the bm ... | 1991 | 1703373 |
| changes in macromolecular synthesis of gypsy moth cell line iplb-ld652y induced by autographa californica nuclear polyhedrosis virus infection. | the aberrant replication of the autographa californica multiple-enveloped nuclear polyhedrosis virus (acmnpv) in the lymantria dispar cell line iplb-ld652y was used as a model system for the investigation of factors regulating baculovirus host specificity. a previous study of this system indicates that viral gene expression in infected cells is extremely attenuated and subsequently all cellular and viral protein synthesis is inhibited. in the present study, infection of iplb-ld652y cells with ac ... | 1991 | 1709681 |
| enzyme activities in four different forms of human immunodeficiency virus 1 pol gene products. | five cassettes of the pol gene of human immunodeficiency virus 1 were constructed and inserted under the control of the polyhedrin gene promoter of autographa californica nuclear polyhedrosis virus by homologous recombination. the first cassette polf contains the full-length pol open reading frame; the second cassette pol100 starts with the first aug codon of the pol gene and deletes 103 amino acids from the amino terminus of the pol gene product; the third cassette pol97 deletes the entire prot ... | 1991 | 1711203 |
| expression of post-translational processing of preprocecropin a using a baculovirus vector. | a cdna fragment encoding preprocecropin a was inserted into the baculovirus autographa californica nuclear polyhedrosis virus downstream of the polyhedrin promoter. the gene was expressed in recombinant-infected last instar larvae of trichoplusia ni and in diapausing pupae of hyalophora cecropia. the identity of the recombinant product was established by electrophoresis with detection of antibacterial activity and mass spectrometry. the prepropeptide had been correctly processed including remova ... | 1991 | 1712729 |
| construction, purification and characterization of a recombinant baculovirus containing the gene for alpha subunit of human chorionic gonadotropin. | a cdna encoding the alpha subunit of human chorionic gonadotropin, a placental glycoprotein hormone, was cloned downstream to the viral polyhedrin gene promoter of autographa california nuclear polyhedrosis virus and the recombinant transfer vector was used to co-transfect spodoptera frugiperda cells growing in culture. recombinant baculovirus carrying the alpha hcg gene was detected and isolated after dot hybridization using supernatant from co-transfected cells. recombinant vac alpha hcg havin ... | 1991 | 1721605 |
| production of recombinant proteins by baculovirus-infected gypsy moth cells. | an experimental study was undertaken to evaluate alternative insect cell lines to sf9 [from spodoptera frugiperda (fall armyworm)] for the production of recombinant proteins. insect cell lines from two different organisms were considered: iplb-ldeita (ldeita) from lymantria dispar (gypsy moth) and iplb-hvt1 (hvt1) from heliothis virescens (tobacco budworm). both ldeita and hvt1 produced higher total activity levels of recombinant beta-galactosidase in monolayer culture than sf9 after infection w ... | 1991 | 1367995 |
| processing, secretion, and immunoreactivity of carboxy terminally truncated dengue-2 virus envelope proteins expressed in insect cells by recombinant baculoviruses. | two recombinant baculoviruses were constructed by inserting via the transfer vector pacym1 the genes coding for the structural proteins of dengue (den)-2 virus downstream from the polyhedrin promoter of autographa californica nuclear polyhedrosis virus. the two recombinants differed in truncation of 26 and 71 amino acids, respectively, in the carboxy-terminal sequence of den-specific envelope (e) glycoprotein. recombinant den-2 e glycoproteins were processed and transported to the surface of spo ... | 1991 | 1984665 |
| proteins encoded in the 81.2- to 85.0-map-unit fragment of autographa californica nuclear polyhedrosis virus dna can be translated in vitro and in spodoptera frugiperda cells. | we have previously demonstrated that five open reading frames exist in the nucleotide sequence of the 81.2- to 85.0-map-unit (m.u.) segment of plaque isolate e of autographa californica nuclear polyhedrosis virus (acnpv) dna. the corresponding polypeptides are 9.8, 12.1, 36.6, 25.0, and 48.2 kda in size (c. oellig, b. happ, t. müller, and w. doerfler, j. virol. 63:1494, 1989), and we have investigated whether these proteins can be translated in infected cells. on subfragments of this viral dna s ... | 1991 | 1985220 |
| identification of the very early transcribed baculovirus gene pe-38. | we have started to identify early viral rnas that are transcribed at 1 h after inoculation to investigate the mechanism involved in the regulation of early gene expression of autographa californica nuclear polyhedrosis virus (acnpv). cloned viral dna fragments were hybridized to northern (rna) blots of polyadenylated rna isolated from spodoptera frugiperda cells at 1, 2, and 6 h postinfection to localize very early transcripts. subsequently we prepared a cdna library of polyadenylated rna transc ... | 1991 | 1987375 |
| transient expression of the autographa californica nuclear polyhedrosis virus immediate-early gene, ie-n, is regulated by three viral elements. | autographa californica nuclear polyhedrosis virus (acmnpv) is a double-stranded dna virus that expresses several immediate-early genes under the control of different promoters. the expression of one of these transcription units, ie-n, is shown here, by a transient expression assay, to be regulated by both cis- and trans-acting viral elements. the steady-state levels of ie-n mrna were very abundant soon after infection but were nearly undetectable during the late phase of the viral life cycle. an ... | 1991 | 1987380 |
| identification and characterization of the ie-1 gene of orgyia pseudotsugata multicapsid nuclear polyhedrosis virus. | the ie-1 gene of orgyia pseudotsugata multicapsid nuclear polyhedrosis virus (opmnpv) was mapped between 95.7 and 97.1 map units on the viral genome. sequence analysis of the opmnpv ie-1 gene (opie-1) identified an open reading frame that coded for a predicted protein of 560 amino acids with a molecular weight of 64,775. transcriptional analysis of opmnpv-infected lymantria dispar (ld652y) cells identified two rnas homologous to the opie-1 open reading frame that were 1.7 and 1.9 kb in size. the ... | 1991 | 1989381 |
| immunoelectron microscopic examination of orgyia pseudotsugata multicapsid nuclear polyhedrosis virus-infected lymantria dispar cells: time course and localization of major polyhedron-associated proteins. | immunoelectron microscopy was employed to examine the temporal expression and localization of two proteins involved in baculovirus polyhedron assembly (polyhedrin and p10) of orgyia pseudotsugata multicapsid nuclear polyhedrosis virus (opmnpv) in infected lymantria dispar cells. in addition, the association of p10 with the polyhedron envelope (pe) protein was studied. the major capsid protein (p39) was also examined to investigate the association of virion structural proteins with polyhedron for ... | 1991 | 1993872 |
| nucleotide sequence and transcript mapping of the hindiii f region of the autographa californica nuclear polyhedrosis virus genome. | the organization of genes in the 2.95 kb ecori-sali fragment (0.7 to 3.0 map units) located within the hindiii f region of the autographa californica nuclear polyhedrosis virus genome was studied by a combination of dna sequencing, northern blot analysis, s1 mapping and primer extension analysis. in addition to the two divergent overlapping transcripts [leftward early (es1) and rightward late (es2)] previously reported, a third transcript which is present at 18 h p.i. and which also runs leftwar ... | 1991 | 1993873 |
| nucleotide sequence of a gene essential for viral dna replication in the baculovirus autographa californica nuclear polyhedrosis virus. | the nucleotide sequence of the 60.1- to 65.5-m.u. region of autographa californica nuclear polyhedrosis virus (acmnpv) was determined. seven large open reading frames were identified. two open reading frames potentially encoding gene products of 143 and 38 kda were found in the counterclockwise direction upstream of the p6.9 gene. four additional open reading frames were found in the opposite direction. analysis of the predicted amino acid sequence of the 143-kda gene revealed a potential leucin ... | 1991 | 1994581 |
| characterization and purification of a functional rat glucocorticoid receptor overexpressed in a baculovirus system. | the structure-function relationship of the oligomeric unactivated glucocorticoid receptor is not fully understood. an essential step in the process of understanding such a relationship involves the production of large quantities of the receptor. using a baculovirus expression system we have been able to overproduce a recombinant rat glucocorticoid receptor (rgr). a cdna coding for the entire rgr was introduced into the genome of the wild type baculovirus, autographa californica nuclear polyhedro ... | 1991 | 1995640 |
| sequential rearrangement and nuclear polymerization of actin in baculovirus-infected spodoptera frugiperda cells. | proper assembly of nucleocapsids of the baculovirus autographa californica nuclear polyhedrosis virus is prevented by cytochalasin d, a drug that interferes with actin microfilament function. to investigate the involvement of microfilaments in a. californica nuclear polyhedrosis virus replication, a fluorescence microscopy study was conducted that correlated changes in distribution of microfilaments with events in the life cycle of the virus. tetramethylrhodamine isothiocyanate-labeled phalloidi ... | 1991 | 1995943 |
| the influence of antisense rna on transcriptional mapping of the 5' terminus of a baculovirus rna. | s1 nuclease protection and primer extension analyses were used to determine the 5' end of the autographa californica nuclear polyhedrosis virus 603 open reading frame (orf) transcript upstream of and on the opposite strand to the polyhedrin gene. these analyses suggested that the 5' end of the 603 orf was located near the initiation site for polyhedrin gene transcription. primer extension products of reverse transcription of 603 rna were dependent on the presence of the taag sequence, an essenti ... | 1991 | 2005431 |
| enhanced secretion from insect cells of a foreign protein fused to the honeybee melittin signal peptide. | the baculovirus/insect cell system has been remarkably successful in yielding high levels of synthesis of many proteins which have been difficult to synthesize in other host/vector systems. the system is also capable of secreting heterologous proteins, but with generally low efficiency. we have increased the efficiency of secretion of the system by using signal peptides of insect origin to direct the secretion of a foreign protein. the precursor of the plant cysteine protease papain (propapain) ... | 1991 | 2016060 |
| characterization of v-sis protein expressed in silkworm larvae using the bombyx mori nuclear polyhedrosis virus vector. | the v-sis oncogene of simian sarcoma virus encodes a protein which is homologous to the human platelet-derived growth factor b-chain. the v-sis protein undergoes a series of processing steps including dimer formation and proteolytic digestion to generate several molecular sizes of the protein. two of these v-sis proteins were expressed alone or as polyhedrin-sis fusion proteins using the bombyx mori nuclear polyhedrosis virus vector. the polyhedrin-sis fusion proteins contained a collagenase-sen ... | 1991 | 2016272 |
| baculovirus-directed high level expression of the hepatitis delta antigen in spodoptera frugiperda cells. | the hepatitis delta antigen (hdag) is a multifunctional protein. it forms the core-like structure of the hepatitis delta virus (hdv) but also enhances replication of hdv in the nucleus of the hepatocyte. a cdna fragment encoding hdag was inserted adjacent to the polyhedrin promoter of autographa californica nuclear polyhedrosis virus present in the baculovirus transfer vector pvl941. after transfection of spodoptera frugiperda (sf9) cells a recombinant baculovirus ac delta 1 was isolated and pur ... | 1991 | 2016595 |
| replication of heliothis zea nuclear polyhedrosis virus in cloned cell lines. | clones from two heliothis zea (lepidoptera:noctuidae) ovarian cell lines, bcirl-hz-am1 (am1) and bcirl-hz-am3 (am3), were generated and their ability to produce h. zea nuclear polyhedrosis virus (hzsnpv) was compared. titers of extracellular virus (ecv) ranged from 5.5 (am3-f9) to 44.9 x 10(5) pfu/ml (am1-a4), with the parental cell lines am3 and am1 producing 14.8 and 26.4 x 10(5) pfu/ml, respectively. concentrations of polyhedral inclusion bodies (pib) produced by the cloned lines ranged from ... | 1991 | 2022870 |
| molecular analysis of a baculovirus regulatory gene. | to better understand the structure and function of a baculovirus regulatory gene, the nucleotide sequence of ie-n expressed by autographa californica nuclear polyhedrosis virus was determined. the 2.0-kb psti-ecorv restriction fragment (97.5 to 98.9 mu) encodes the upstream regulatory sequences, open reading frame, and downstream sequences of the immediate early ie-n gene. using a convenient restriction site, the 285-bp promoter of ie-n was divided into two functional regions as defined by trans ... | 1991 | 2024466 |
| autographa californica m nuclear polyhedrosis virus and cytochalasin d: antagonists in the regulation of protein synthesis. | during infection by the baculovirus autographa californica multiple nuclear polyhedrosis virus (acmnpv), synthesis of host proteins in iplb-sf-21 cells is inhibited. we report here that cytochalasin d (cd), a drug that specifically interacts with actin, behaved as an antagonist in the virus-mediated shutdown of host proteins actin and tubulin. in uninfected cells, cd caused an increase in actin synthesis but had no apparent effect on tubulin synthesis. in infected cells, cd similarly enhanced ac ... | 1991 | 2024491 |
| expression and cell membrane localization of rat m3 muscarinic acetylcholine receptor produced in sf9 insect cells using the baculovirus system. | a recombinant baculovirus bearing the cdna coding for the rat muscarinic acetylcholine receptor subtype m3 placed under the control of the autographa californica nuclear polyhedrosis virus polyhedrin gene promoter, was constructed. polymerase chain reaction screening was used to identify the recombinant baculovirus. northern blot analysis of total rna from insect cells infected with the recombinant baculovirus indicated that the transcripts were abundant. binding assays carried out with the musc ... | 1991 | 2037073 |
| host range expansion by recombination of the baculoviruses bombyx mori nuclear polyhedrosis virus and autographa californica nuclear polyhedrosis virus. | the mechanisms of host specificity of nuclear polyhedrosis viruses (npvs) (baculoviridae) were analyzed after coinfection of bombyx mori npv (bmnpv) and one of four distinct groups of spodoptera litura npv (slnpv), including an autographa californica npv (acnpv) variant (s. maeda, y. mukohara, and a. kondo, j. gen. virol. 71:2631-2639, 1990), into various lepidopteran cell lines. replication of bmnpv in nonpermissive cells (tn-386, sf-21, and cls-79) was induced by coinfection with acnpv but not ... | 1991 | 2041087 |
| expression of an enhancer-binding protein in insect cells transfected with the autographa californica nuclear polyhedrosis virus ie1 gene. | the baculovirus autographa californica nuclear polyhedrosis virus contains an element known as homologous region 5 (hr5) which is an enhancer of delayed-early viral gene expression. to begin to identify proteins that interact with hr5, dna-protein interactions were analyzed by using extracts from spodoptera frugiperda cells and a fragment of dna containing the left half of the hr5 enhancer. this 252-bp dna fragment contains two copies of a 30-bp direct repeat (dr30) and two copies of a 24-bp imp ... | 1991 | 2041088 |
| baculovirus vectors for multiple gene expression and for occluded virus production. | the nature and properties of a variety of plasmids are described that facilitate the construction of baculovirus vectors for expression of one or more heterologous genes. the plasmids are designed for use with autographa californica nuclear polyhedrosis virus, acmnpv, as a vector for protein production in insect cells and/or insect larvae. several plasmids described here facilitate the simultaneous insertion and expression of two different genes. some vector systems allow high and equal levels o ... | 1991 | 2055465 |
| expression of the x protein of hepatitis b virus in insect cells using recombinant baculoviruses. | the baculovirus system was used to express the x protein of human hepatitis b virus (hbv). the x open reading frames (x orfs) from cloned viral dna of the hbv subtypes ayw and adr were introduced into the genome of autographa californica nuclear polyhedrosis virus (acnpv). the hbv-dna of subtype adr derived from a hepatocellular carcinoma contains an x orf and a 5' extended prex/x orf, which were both used to construct x recombinant baculoviruses. infection of sf9 insect cells with these recombi ... | 1991 | 2058090 |
| construction of an improved baculovirus insecticide containing an insect-specific toxin gene. | baculoviruses provide alternatives to chemicals for controlling insect pests and can be applied by spraying. baculoviruses have a limited host range, but work relatively slowly. they are dissolved in the midgut of insect larvae to release infectious virions which enter gut epithelial cells and begin to replicate. replication in other organs causes extensive tissue damage and eventually death. this process can take 4-5 days, but in the field may last for more than a week, allowing the larvae to f ... | 1991 | 2062383 |
| high-level expression of a mitochondrial enzyme, ornithine transcarbamylase from rat liver, in a baculovirus expression system. | the mitochondrial enzyme, ornithine transcarbamylase (otc) from rat liver was expressed in spodoptera frugiperda (sf) insect cells using a baculovirus vector. when insect cells were infected with recombinant autographica californica nuclear polyhedrosis virus (acnpv) containing a cdna encoding the precursor form of otc (potc) inserted into the polyhedrin gene, they expressed catalytically active enzyme at levels of approximately 2.5 micrograms/10(6) cells. about 25% of the active enzyme was a no ... | 1991 | 2069719 |
| identification of upstream promoter elements mediating early transcription from the 35,000-molecular-weight protein gene of autographa californica nuclear polyhedrosis virus. | site-directed mutagenesis was used to examine the organization of cis-acting regulatory elements that comprise the promoter of the early 35,000-molecular-weight protein gene (35k protein gene) encoded by the ecori-s region of the baculovirus autographa californica nuclear polyhedrosis virus. the promoter fragment, extending from positions -226 to +12 relative to the early rna start site (position +1), was fused to the reporter gene encoding chloramphenicol acetyltransferase (cat) and then insert ... | 1991 | 2072443 |
| expression and characterization of recombinant mouse beta 2-microglobulin type a in insect cells infected with recombinant baculoviruses. | the murine beta 2-microglobulina cdna was cloned into pac373 and pvl941 transfer vectors and introduced via homologous recombination into the genome of autographa californica nuclear polyhedrosis virus downstream of the polyhedrin promoter. both types of recombinant baculoviruses were isolated and used to infect spodoptera frugiperda (sf9) lepidopteran cells. beta 2m was synthesized at a substantially higher rate in cells infected with the pvl941-derived virus than when the pac373-based virus wa ... | 1991 | 1754712 |
| effect of an autographa californica nuclear polyhedrosis virion component(s) on dna synthesis and growth in several insect cell lines. | thirteen different insect cell lines representing three different orders were infected with autographa californica nuclear polyhedrosis virus (acmnpv) whose genome had been inactivated in situ by photochemical means or by short wave uv irradiation. changes in rates of cellular dna synthesis, as measured by [3h]thymidine incorporation, and cell growth were subsequently measured at various times post infection. seven cell lines exhibited a significant decline in [3h]thymidine incorporation (compar ... | 1991 | 1759911 |
| a baculovirus dual expression vector derived from the autographa californica nuclear polyhedrosis virus polyhedrin and p10 promoters: co-expression of two influenza virus genes in insect cells. | a baculovirus transfer vector, pacuw3, was developed to facilitate the insertion of two influenza virus genes, those encoding the haemagglutinin (ha) and neuraminidase (na) membrane glycoproteins, into the autographa californica nuclear polyhedrosis virus genome in a single cotransfection experiment. the na gene was inserted in place of the polyhedrin coding sequences under the control of the polyhedrin promoter, whereas the ha gene was placed under the control of a copy of the p10 promoter at a ... | 1991 | 1765769 |
| structural comparison of the autographa californica nuclear polyhedrosis virus-induced rna polymerase and the three nuclear rna polymerases from the host, spodoptera frugiperda. | a partial subunit structure has been determined for the novel rna polymerase that is induced in fall armyworm (spodoptera frugiperda) cells upon infection with the autographa californica nuclear polyhedrosis virus (acnpv). the putative structure includes nine polypeptides; the complexity of this structure is in accord with the high sedimentation coefficient (15s) estimated for this enzyme. a comparison of the putative structure of the virus-induced polymerase with those of the three host nuclear ... | 1991 | 1767583 |
| expression of the s1 sub-unit of pertussis toxin using a recombinant baculovirus. | the full-length genome of the s1 sub-unit containing the native signal sequence was inserted downstream of the polyhedrin promoter of autographa californica nuclear polyhedrosis virus. a large amount of 28 kd protein, which was larger than the authentic s1 sub-unit (26 kd), was detected in insect cells infected with the recombinant virus by coomassie blue staining after resolving by sds-page. although the level of expression of the recombinant s1 sub-unit was high, immunoblotting and n-terminal ... | 1991 | 1778337 |
| nucleotide sequence of the gene coding for p40, an occluded virion-specific polypeptide of bombyx mori nuclear polyhedrosis virus. | 1991 | 1783783 | |
| high-level expression and characterization of hepatitis b virus surface antigen in silkworm using a baculovirus vector. | the bombyx mori nuclear polyhedrosis virus (bmnpv) in the silkworm was used successfully for mass production of biologically active foreign genes under the control of the polyhedrin promoter. this system was adapted for the production of large amounts of hepatitis b virus surface antigens (hbsag). the dna fragments coded for the middle protein, which is composed of the s protein with the pre-s2 region, were cloned, the signal protein gene of beta-ifn was added, and both were inserted into a clon ... | 1991 | 1816252 |
| the nonstructural protein (nss) encoded by the ambisense s rna segment of tomato spotted wilt virus is associated with fibrous structures in infected plant cells. | the open reading frame located in the viral strand of the ambisense s rna of tomato spotted wilt virus (tswv), was cloned into transfer vector pac33dz1 and inserted downstream of the polyhedrin promoter in the autographa californica nuclear polyhedrosis virus genome. recombinant baculoviruses were obtained that showed a high-level expression of a 52.4-kda protein corresponding to the inserted tswv gene. the viral protein thus produced was purified and injected into rabbits to raise antibodies. w ... | 1991 | 1826573 |
| secretion of single-chain urokinase-type plasminogen activator from insect cells. | a cdna encoding human urokinase-type plasminogen activator was inserted downstream from the polyhedrin promoter of the baculovirus autographa californica nuclear polyhedrosis virus. a protein of similar mr to urokinase (uk) was synthesized and approx. 90% was secreted from recombinant virus-infected spodoptera frugiperda cells. zymography and western blotting analysis of the insect-derived protein demonstrated that it was comprised solely of the high-mr form of uk. no low-mr uk was detected. ami ... | 1991 | 1834526 |
| field evaluation of a dna hybridization assay for nuclear polyhedrosis virus in gypsy moth (lepidoptera: lymantriidae) larvae. | dna hybridization assays were used to detect the presence of viral dna in gypsy moth (lymantria dispar l.) larvae collected weekly from high density populations or reared from field-collected egg masses. dna was extracted from larvae, bound to nitrocellulose filters, and hybridized with digoxigenin-labeled l. dispar npv (ldnpv) dna probes. the virus incidence determined from dna hybridization assays was compared with that determined with conventional microscopic examination of larvae for polyhed ... | 1991 | 1842802 |
| high-level expression of the epstein-barr virus alkaline deoxyribonuclease using a recombinant baculovirus: application to the diagnosis of nasopharyngeal carcinoma. | the epstein-barr virus (ebv) alkaline deoxyribonuclease (dnase) was inserted into the baculovirus autographa californica nuclear polyhedrosis virus (acmnpv). infection of the insect cell line spodoptera frugiperda (sf9) with the recombinant virus led to the expression of an enzymatically active alkaline dnase. the recombinant ebv alkaline dnase was highly soluble, and the recombinant baculovirus produced approximately 10-20 mg of ebv dnase per 1 x 10(9) cells. the recombinant enzyme activity was ... | 1991 | 1847261 |
| direct double-stranded dna sequencing with baculovirus genomes. | double-stranded dna sequencing with the modified t7 dna polymerase (sequenase) was performed directly with nuclear polyhedrosis virus dna genomes. the conditions were optimized to allow for a rapid and unambiguous sequence analysis of nuclear polyhedrosis virus genomes. | 1991 | 1849913 |
| detection and analysis of autographa californica nuclear polyhedrosis virus mutants with defective interfering properties. | defective interfering particles (dips) were generated upon continuous production of autographa californica nuclear polyhedrosis virus (acnpv) in bioreactors. this configuration mimicked the serial undiluted passaging of virus, which is known to result in plaque-morphology mutants. restriction enzyme analysis of dip-containing preparations of extracellular virus showed the presence of many dna fragments in less than equimolar amounts. these fragments were colinear on the physical map of acnpv and ... | 1991 | 1853572 |
| a baculovirus homolog of a cu/zn superoxide dismutase gene. | a gene with greater than 50% amino acid sequence identity to eukaryotic cu/zn superoxide dismutase genes (sod) was found at 19 map units in the genome of the baculovirus autographa californica nuclear polyhedrosis virus (acmnpv). this gene was transcribed into 1.4- and 1.5-kb rnas late in virus infection. the two late rnas have coterminal 3' ends but initiate from two different start points, both of which map to the central adenine of the sequence motif attaag. the late gene product was found to ... | 1991 | 1871962 |
| dengue-1 virus envelope glycoprotein gene expressed in recombinant baculovirus elicits virus-neutralizing antibody in mice and protects them from virus challenge. | in order to test the feasibility of baculovirus (autographa californica nuclear polyhedrosis virus, acnpv) expression vectors for making immunogens against dengue-1 (den-1) virus, a portion of the envelope (e) glycoprotein gene of den-1 virus was cloned and expressed. the recombinant baculovirus contains 107 nucleotides from the 3' terminus of the den-1 matrix (m) gene, which encodes a hydrophobic signal peptide and extends through the first 1, 245 nucleotides of e, terminating 243 nucleotides b ... | 1991 | 1877710 |
| synthesis of the virus-specified tubules of epizootic haemorrhagic disease virus using a baculovirus expression system. | the formation of virus-specific tubules is one of the most characteristic features in the orbivirus infection cycle, yet little is known about their role in virus replication. the tubuli are composed of a major nonstructural protein, ns1. we have investigated the expression of the ns1-encoding gene of epizootic haemorrhagic disease virus serotype 2 (alberta-strain) by producing a recombinant autographa californica nuclear polyhedrosis virus (acnpv). prior to cloning in the baculovirus transfer v ... | 1991 | 1891959 |
| repeat sequences from complex ds dna viruses can be used as minisatellite probes for dna fingerprinting. | in a search for new fingerprinting probes for use with sheep, repeat sequences derived from five poxviruses, an iridovirus and a baculovirus were screened against dna from sheep pedigrees. probes constructed from portions of the parapox viruses, orf virus and papular stomatitis virus and the baculovirus from the alfalfa looper, autographa californica, nuclear polyhedrosis virus all gave fingerprint patterns. probes from three other poxviruses and an iridovirus did not give useful banding pattern ... | 1991 | 1892249 |
| novel regulatory properties of the ie1 and ie0 transactivators encoded by the baculovirus autographa californica multicapsid nuclear polyhedrosis virus. | the baculovirus autographa californica multicapsid nuclear polyhedrosis virus expresses two immediate-early genes from the hindiii-g region (map units 90.4 to 96.8) of the genome. during the early phase of infection, nonspliced 1.9-kb and spliced 2.1-kb transcripts are expressed which encode the ie1 and ie0 (spliced ie1) gene products, respectively. these two gene products differ only in that ie0 contains an additional 54 amino acids at the amino terminus. rna analysis of these two genes during ... | 1991 | 1895384 |
| expression of human alzheimer amyloid precursor protein in insect cells. | the amyloid beta-peptide is a major constituent of amyloid deposited in the brains of patients with alzheimer's disease and is derived from a larger precursor protein/s (app-695, 751, 770). a human cdna encoding full-length app-751 was inserted into the genome of autographa californica nuclear polyhedrosis virus under transcriptional regulation of the viral polyhedrin gene promoter. the recombinant virus was used to infect insect cells, which resulted in the abundant expression of app-751. analy ... | 1991 | 1899577 |
| synthesis of biologically active adenovirus preterminal protein in insect cells using a baculovirus vector. | a dna fragment encoding the polyhedrin promoter of autographa californica multiple nuclear polyhedrosis virus (acmnpv strain) was constructed using overlapping oligodeoxyribonucleotides (oligos), which included the 5'-untranslated leader sequence of the polyhedrin-encoding gene. this dna fragment was cloned into an intermediate transfer vector (pkx105) providing a unique bamhi site for the insertion of foreign genes. the escherichia coli lacz gene was first cloned at the bamhi site of pkx105 and ... | 1991 | 1905255 |
| alpha-mannosidase-catalyzed trimming of high-mannose glycans in noninfected and baculovirus-infected spodoptera frugiperda cells (iplb-sf-21ae). a possible contributing regulatory mechanism for assembly of complex-type oligosaccharides in infected cells. | incubation of a spodoptera frugiperda (iplb-sf-21ae) cell extract with the oligosaccharide man9glcnac2, the aglucosyl derivative of the glycan that is normally transferred from the dolichol carrier to the relevant asn residue in the nascent protein, results in its trimming to man6glcnac2, an intermediate that is relatively stable to further alpha-d-mannosidase action in these cells. on the other hand, incubation of a similar extract from cells that had been infected for various times with a wild ... | 1991 | 1911772 |
| transcriptional analyses of baculovirus polyhedrin and foreign gene expression relative to baculovirus p10 mrna levels. | comparisons have been made between the p10 and polyhedrin mrna levels recovered from spodoptera frugiperda cells infected with autographa californica nuclear polyhedrosis virus (acnpv). in molar terms and from 18 h post-infection (p.i.), the polyhedrin mrna species increased to levels one and a half times to twice as high as the p10 levels. the influence of the polyhedrin leader sequence on the expression of a foreign gene under the control of the polyhedrin promoter was investigated using a ser ... | 1991 | 1919531 |
| baculovirus gp64 gene expression: analysis of sequences modulating early transcription and transactivation by ie1. | expression of the baculovirus major envelope glycoprotein gene (gp64) is regulated by transcription from both early and late promoters. to characterize the early promoter and identify sequences involved in the regulation of gp64 early transcription, promoter-reporter gene fusions were generated from the orygia pseudotsugata nuclear polyhedrosis virus gp64 promoter and were analyzed by transient expression in uninfected insect cells. for these analyses, 5' deletion mutations were constructed in t ... | 1991 | 1920618 |