Publications

a baculovirus expression vector derived from the basic protein promoter of autographa californica nuclear polyhedrosis virus.	the basic protein of autographa californica nuclear polyhedrosis virus (acmnpv) is associated with virus dna in virion nucleocapsids and is produced in infected cells during the late phase of gene expression. a transfer vector was constructed containing the beta-galactosidase gene, under the control of a copy of the putative basic protein promoter, in place of the polyhedrin gene within the acmnpv ecori fragment i. after cotransfection of spodoptera frugiperda cells with the transfer vector and ...	1990	2109042
localization of the 34 kda polyhedron envelope protein in spodoptera frugiperda cells infected with autographa californica nuclear polyhedrosis virus.	using immuno-electron microscopy the 34 kda polyhedron envelope (pe) phosphoprotein (pp 34) was localized in cells infected with autographa californica multiple-nucleocapsid nuclear polyhedrosis virus (acmnpv). in wild-type acmnpv-infected cells this protein was found associated with electron-dense "spacers" and the polyhedron envelopes demonstrating their structural relationship. in these cells pp 34 was also found associated with fibrillar structures present in the nucleus and cytoplasm of inf ...	1990	2109589
human acid beta-glucosidase: glycosylation is required for catalytic activity.	the role of oligosaccharide modification in human acid beta-glucosidase function was investigated. this lysosomal enzyme has five putative n-glycosylation sites, four of which are occupied. the unglycosylated human protein was stable when expressed in bacteria or in spodoptera frugiperda cells in the presence of tunicamycin but lacked catalytic activity. deglycosylation of purified acid beta-glucosidase from human placenta with n-glycanase under native conditions resulted in the removal of an ac ...	1990	2110456
expression and characterization of a functional human immunodeficiency virus envelope glycoprotein in insect cells.	recombinant baculoviruses were used to express the gp160 envelope glycoprotein of the human immunodeficiency virus type 1 (hiv-1) and a truncated variant designated gp160(t) which lacks a transmembrane domain. glycosylation, proteolytic cleavage, secretion, and biological activities of gp160 and gp160(t) 160(t) were studied in spodoptera frugiperda cells. both proteins were rapidly glycosylated and initially were found to be totally endo-beta-n-acetyl-d-glucosaminidase h (endo-h) sensitive. howe ...	1990	2111957
cloning and expression of human aldose reductase.	the complete amino acid sequence of human retina and muscle aldose reductase was determined by nucleotide analysis of cdna clones isolated using synthetic oligonucleotide probes based on partial amino acid sequences of purified human psoas muscle aldose reductase. the cdna sequence differs substantially in the noncoding and coding regions of recently published sequences of this enzyme. the mrna for aldose reductase was abundantly expressed in hela cells, but only scarcely in a neuroblastoma cell ...	1990	2112546
alpha 1----3-galactosyltransferase: the use of recombinant enzyme for the synthesis of alpha-galactosylated glycoconjugates.	we have reported the isolation and characterization of a bovine cdna clone containing the complete coding sequence for udp-gal:gal beta 1----4glcnac alpha 1----3-galactosyltransferase [joziasse, d. h., shaper, j. h., van den eijnden, d. h., van tunen, a. j. & shaper, n. l. (1989) j. biol. chem. 264, 14290-14297]. insertion of this cdna clone into the genome of autographa californica nuclear polyhedrosis virus (acnpv) and subsequent infection of spodoptera frugiperda (sf9) insect cells with recom ...	1990	2116309
cell cycle kinetics of insect cell cultures compared to mammalian cell cultures.	cell cycle kinetic of lepidopteran cell lines sf9 (spodoptera frugiperda) and izdmb0503 (mamestra brassicae) were investigated and compared to mammalian cell cycle distributions. the resting phase (g0) of mammalian cells is characterized by a 2c-dna content whereas g0-phase of insect cell lines is characterized by a 4c-dna content. flow cytometric data in combination with growth curves of partially synchronized and asynchronously growing cells proved the existence of this phenomenon. kinetics of ...	1990	2142463
co-expression of the human hla-b27 class i antigen and the e3/19k protein of adenovirus-2 in insect cells using a baculovirus vector.	we have expressed the human mhc class i hla-b27 antigen, human beta 2-microglobulin, and the e3/19k protein of adenovirus-2 in spodoptera frugiperda insect cells (sf9) by using the autographa californica nuclear polyhedrosis virus. all genes were inserted under the strong polyhedrin promoter in the vector pvl941. the proteins were expressed at high levels, ranging from 1 to 8 mg protein per 3 x 10(9) cells. both a full-length and a truncated form of hla-b27 were expressed. the latter was termina ...	1990	2150332
identification of bluetongue virus vp6 protein as a nucleic acid-binding protein and the localization of vp6 in virus-infected vertebrate cells.	recently the insect baculovirus autographa californica nuclear polyhedrosis virus (acnpv) has been effectively adapted as a highly efficient vector in insect cells for the expression of various genes. a cdna sequence of rna segment 9 of bluetongue virus serotype 10 (btv-10, an orbivirus member of the reoviridae family) encoding a minor core protein (vp6) has been inserted into the bamhi site of the pacym1 transfer vector derived from acnpv. spodoptera frugiperda cells were cotransfected with the ...	1990	2152806
identification of a viral gene encoding a ubiquitin-like protein.	the baculovirus autographa californica nuclear polyhedrosis virus (acmnpv, which is representative of the mnpv subtype in which the virions may contain many nucleocapsids within a single viral envelope) encodes a protein, v-ubi, that has 76% identity with the eukaryotic protein ubiquitin. transcriptional mapping indicated that the gene for v-ubi was transcribed during the late phase of viral infection. two transcriptional start sites potentially encoding v-ubi were identified. both sites were co ...	1990	2153300
sequence comparison of cellular and viral copies of host cell dna insertions found in autographa californica nuclear polyhedrosis virus.	the nucleotide sequence and structural characteristics of two nonhomologous host dna insertions of spodoptera frugiperda (fall armyworm) origin isolated from few polyhedra mutants of the baculovirus autographa californica nuclear polyhedrosis virus (acmnpv) have been determined. neither of the host insertions contain long open reading frames suggesting that cellular genes were not introduced into the viral genome. one of the host dna insertions, ifp1.6, terminated in short imperfect inverted rep ...	1990	2154879
purification and characterization of the recombinant extracellular domain of human nerve growth factor receptor expressed in a baculovirus system.	to obtain the large quantities of the extracellular domain of the nerve growth factor receptor (ngf-r) necessary for structural analyses, we produced this protein in the baculovirus expression system. a cdna coding for the extracellular domain of the human ngf-r was first introduced into transfer vector pvl941. recombinant baculovirus was produced by cotransfecting spodoptera frugiperda cells with the transfer vector and dna of autographa californica nuclear polyhedrosis virus. recombinant viral ...	1990	2155239
synthesis of bluetongue virus (btv) corelike particles by a recombinant baculovirus expressing the two major structural core proteins of btv.	the l3 and m7 genes of bluetongue virus (btv), which encode the two major core proteins of the virus (vp3 and vp7, respectively), were inserted into a baculovirus dual-expression transfer vector and a recombinant baculovirus expressing both foreign genes isolated following in vivo recombination with wild-type autographa californica nuclear polyhedrosis virus dna. spodoptera frugiperda insect cells infected with the recombinant synthesized large amounts of btv corelike particles. these particles ...	1990	2157041
an insertion of insect cell dna in the 81-map-unit segment of autographa californica nuclear polyhedrosis virus dna.	in this report, a transposonlike insertion of spodoptera frugiperda insect cell dna was analyzed in single-plaque isolate e of the insect baculovirus autographa californica nuclear polyhedrosis virus (acnpv). the 634-base-pair insertion is characterized by an 18-base-pair terminal inverted repeat and carries an ecori site. this additional ecori site in the 81-map-unit segment of the dna of plaque isolate e of acnpv explains the difference between the ecori restriction map of the dna from this is ...	1990	2157067
enzymatic activities of overexpressed herpes simplex virus dna polymerase purified from recombinant baculovirus-infected insect cells.	biochemical characterization of the herpes simplex virus (hsv) dna polymerase, a model dna polymerase and an important target for antiviral drugs, has been limited by a lack of pure enzyme in sufficient quantity. to overcome this limitation, the hsv dna polymerase gene was introduced into the baculovirus, autographa californica nuclear polyhedrosis virus, under the control of the polyhedrin promoter to give rise to a recombinant baculovirus, bp58. bp58-infected spodoptera frugiperda insect cells ...	1990	2157192
protection of mice against lethal japanese encephalitis with a recombinant baculovirus vaccine.	genes coding for the e and ns1 glycoproteins of japanese encephalitis virus (jev) were cloned into baculovirus expression vectors. the recombinant baculoviruses obtained were used to infect spodoptera frugiperda cells. the infected cells were used to immunize c57/b mice, which were then challenged with live jev. survival was increased from about 30% in unimmunized mice to 70% in e and polyprotein recipients (p less than 0.005), but was not increased in ns1 recipients despite the development of a ...	1990	2160199
high level expression of the two outer capsid proteins of bluetongue virus serotype 10: their relationship with the neutralization of virus infection.	dna representing rna segments 2 and 5 of bluetongue virus (btv) serotype 10, corresponding to the genes that code for the outer capsid proteins vp2 and vp5, have been inserted into a baculovirus transfer vector in lieu of the coding region of the polyhedrin gene of autographa californica nuclear polyhedrosis virus (acnpv). after co-transfection of spodoptera frugiperda cells with wild-type acnpv dna in the presence of the recombinant transfer vector dnas polyhedrin-negative recombinant baculovir ...	1990	2160763
expression of pseudomonas phosphotriesterase activity in the fall armyworm confers resistance to insecticides.	the gene encoding for the phosphotriesterase (opd) from pseudomonas diminuta has been subcloned into a baculovirus expression system. functional enzyme is produced when the recombinant baculovirus is used to infect either cultured spodoptera frugiperda sf9 cells or the larval stage of the fall armyworm. the ld50 for paraoxon toxicity was found to increase 280-fold in the larvae after infection with the recombinant baculovirus and expression of the functional phosphotriesterase.	1990	2164956
biophysical studies on the morphology of baculovirus-expressed bluetongue virus tubules.	bluetongue virus tubules were purified from spodoptera frugiperda cells infected with a recombinant baculovirus containing the ns1 gene from bluetongue virus serotype 10, and expressed under control of the autographa californica nuclear polyhedrosis virus polyhedrin promoter. these tubules were subjected to a variety of chemical and physical treatments and the resulting effects on tubule morphology were examined by electron microscopy. a number of morphological similarities were noted between bl ...	1990	2167940
synthesis of bluetongue virus-encoded phosphoprotein and formation of inclusion bodies by recombinant baculovirus in insect cells: it binds the single-stranded rna species.	a dna clone of rna segment 8 (s8) of bluetongue virus type 10 (btv-10), an orbivirus member of the reoviridae family has been expressed to high levels (20 mg/1 x 10(9) cells) using an autographa californica nuclear polyhedrosis virus expression vector (pa-cym1). the expressed protein is similar to the authentic btv phosphoprotein ns2, in its size, antigenicity, and also the manner of phosphorylation (e.g. same peptides and residues). both mammalian and insect cell-derived ns2 proteins are phosph ...	1990	2170574
expression of avian leukaemia virus env-gp85 in spodoptera frugiperda cells by use of a baculovirus expression vector.	we studied the genetic expression of gp85 of avian leukaemia virus (alv) subgroup a in a baculovirus/insect cell system. 5'terminal sequences of the gag gene were added to precede the alv gp85 sequence and a stop codon was introduced at the boundary of gp85 and gp37. the resulting construct was then cloned into the baculovirus transfer vector pacym1, which contains the polyhedrin promoter of autographa californica nuclear polyhedrosis virus (acnpv). cells of the insect spodoptera frugiperda (sf9 ...	1990	2174957
expression of the full-length products of the human papillomavirus type 6b (hpv-6b) and hpv-11 l2 open reading frames by recombinant baculovirus, and antigenic comparisons with hpv-11 whole virus particles.	the l2 open reading frames (orfs) of human papillomavirus (hpv) types 6b and 11 were expressed as full-length non-fusion proteins in spodoptera frugiperda (sf-9) cells using recombinant baculovirus. both proteins were detected on western blots as immunoreactive bands which migrated with apparent mrs of 76k and 78k, respectively, and contained both cross-reactive and type-specific epitopes, as determined by polyclonal antisera directed against defined subregions of the hpv-6b and hpv-11 l2 orfs. ...	1990	2174962
characterization of the basement membrane glycoprotein entactin synthesized in a baculovirus expression system.	a cdna clone encoding the entire open reading frame of mouse entactin was constructed. this cdna was inserted into autographa californica multiple nuclear polyhedrosis virus and expressed in spodoptera frugiperda, sf9, insect cells. the recombinant entactin was produced in large quantities under the control of the polyhedrin promoter. amino terminus sequence analysis indicated that the signal peptide in the preprotein was correctly cleaved. polyclonal antibodies prepared against either the natur ...	1990	2180961
efficient, low-cost protein factories: expression of human adenosine deaminase in baculovirus-infected insect larvae.	human adenosine deaminase (ec 3.5.4.4), a key purine salvage enzyme essential for immune competence, has been overproduced in spodoptera frugiperda cells and in trichoplusia ni (cabbage looper) larvae infected with recombinant baculovirus. the coding sequence of human adenosine deaminase was recombined into a baculovirus immediately downstream from the strong polyhedrin gene promoter. approximately 60 hr after infection of insect cells with the recombinant virus, maximal levels of intracellular ...	1990	2181448
ecdysteroids increase the yield of recombinant protein produced in baculovirus insect cell expression system.	spodoptera frugiperda cells are the hosts of wild type and recombinant virus in the baculovirus insect cell expression system. the expression of the foreign gene could be enhanced by the addition of ecdysteroids and increased amount of recombinant protein was secreted into the medium. the time and concentration dependence of this effect was followed in the case of 20-hydroxyecdysone-, makisterone and ecdysone. 20-hydroxyecdysone proved to be the most efficient, producing a three fold increase in ...	1990	2182025
antigenic parvovirus b19 coat proteins vp1 and vp2 produced in large quantities in a baculovirus expression system.	two baculovirus expression vectors derived from autographica californica nuclear polyhedrosis virus (acnpv) were prepared containing the complete 2.5 kb coding region for parvovirus b19 coat protein vp1 (acb19vp1l) and the 1.8 kb coding region for vp2 (acb19vp2l), placed under the control of the polyhedrin promoter. the recombinant viruses were used to infect spodoptera frugiperda cells and the proteins expressed were analysed using appropriate antibodies. acb19vp1l-infected cells produced b19 v ...	1990	2188463
characterization of the da26 gene in a hypervariable region of the autographa californica nuclear polyhedrosis virus genome.	a region of the baculovirus autographa californica nuclear polyhedrosis virus genome that is frequently found to be altered after serial passage of the virus in cell culture was characterized. sequence analysis of this region of the genome in wild-type and mutant viruses revealed that some of the mutations affected a 675 bp open reading frame, designated da26. the da26 gene was disrupted both by deletion and by insertion of sequences that resembled transposable elements. northern blot analysis o ...	1990	2189022
rabies diagnostic reagents prepared from a rabies n gene recombinant expressed in baculovirus.	a gene encoding the nucleoprotein (n) of rabies virus was inserted into the genome of the baculovirus autographa californica nuclear polyhedrosis virus. recombinant gene expression was controlled by the strong polyhedrin gene promoter. insect cells (spodoptera frugiperda) infected by a baculovirus recombinant containing the rabies virus n gene produced abundant amounts of a novel 55-kilodalton protein of a size comparable to that of the rabies virus n protein, as demonstrated by polyacrylamide g ...	1990	2191008
expression of influenza a and b virus nucleoprotein antigens in baculovirus.	full-length cdna clones of the nucleoprotein (np) genes of influenza a/ann arbor/6/60 and b/ann arbor/1/86 viruses were constructed from virion rna and subsequently expressed in spodoptera frugiperda (sf9) cells using the baculovirus vector, autographa californica nuclear polyhedrosis virus. western blot analysis of lysates prepared from sf9 cells infected with the recombinant viruses confirmed that the baculovirus-expressed np antigens were reactive with monoclonal antibodies specific for eithe ...	1990	2197370
human acidic fibroblast growth factor overexpressed in insect cells is not secreted into the medium.	we have overexpressed human acidic fibroblast growth factor (afgf) in spodoptera frugiperda sf9 cells from a cdna clone under the control of the promoter of the polyhedrin gene of the baculovirus autographa californica nuclear polyhedrosis virus. a 16.5-kd product was made in recombinant virus-infected cells that specifically reacted in immunoblots with various antibodies prepared against afgf. recombinant afgf was mitogenic for bhk21 cells and its activity was stimulated by heparin. the mechani ...	1990	2200451
secretion of functional papain precursor from insect cells. requirement for n-glycosylation of the pro-region.	the synthetic gene coding for the precursor of the cysteine protease papain (ec 3.4.22.2) has been expressed using the baculovirus/insect cell system. the prepropapain gene was cloned into the transfer vector ipdc125 behind the polyhedrin promoter. the recombinant construct was then incorporated by homologous recombination into the autographa californiaca nuclear polyhedrosis virus genome. the host spodoptera frugiperda sf9 cells infected with the recombinant baculovirus secrete an enzymatically ...	1990	2204628
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
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
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
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
ascovirus infectivity and effects of infection on the growth and development of noctuid larvae.	the infectivity per os and by inoculation of ascoviruses isolated from heliothis zea, spodoptera frugiperda, and trichopulsia ni and the effects of infection by these viruses on host growth and development were studied in the species from which these viruses were originally isolated, or in the case of the h. zea isolate, in h. virescens. mortality caused by all three viral isolates averaged less than 15% in third instars fed doses of 10 to 10(5) viral vesicles per larva incorporated into diet, w ...	1990	2250099
comparative histopathology of three ascovirus isolates in larval noctuids.	the histopathology caused by three ascoviruses isolated respectively from heliothis zea, spodoptera frugiperda, and trichoplusia ni was studied in the host species from which the viruses were isolated originally, or in the case of the h. zea isolate, in h. virescens. in all three isolates, infected cells and virion-containing vesicles in the hemolymph were observed by 3 days postinoculation. the isolates from h. zea and t. ni exhibited relatively broad tissue tropisms infecting the tracheal matr ...	1990	2250100
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
physicochemical analysis of the hepatitis b virus core antigen produced by a baculovirus expression vector.	the hepatitis b virus particle consists of an envelope carrying the surface antigen of the virus and an internal capsid consisting of the core antigen (hbcag). the internal capsid contains the circular, partially dsdna genome and the viral polymerase. empty core particles have been produced in spodoptera frugiperda cells using a recombinant baculovirus vector, ym1ktc, that expresses a 21.4k derivative of the hbcag gene. the particles have been purified to homogeneity by caesium chloride density ...	1990	2254755
analyses of catalytic activity and inhibitor binding of human acid beta-glucosidase by site-directed mutagenesis. identification of residues critical to catalysis and evidence for causality of two ashkenazi jewish gaucher disease type 1 mutations.	analyses of catalytic properties and inhibitor binding were conducted to investigate the molecular basis of active site function of human acid beta-glucosidases (ec 3.2.1.45) expressed from normal and gaucher disease type 1 alleles. comparative studies were conducted with enzymes expressed from natural (spleen and fibroblasts) alleles or from mutagenized cdnas in spodoptera frugiperda (sf9) cells using the baculovirus expression system. mutant cdnas containing thr43 to lys43 (beta-glcthr43----ly ...	1990	2324100
overexpression of full-length human glucocorticoid receptor in spodoptera frugiperda cells using the baculovirus expression vector system.	we have expressed a full-length human glucocorticoid receptor (hgr) in spodoptera frugiperda (sf9) cells using the baculovirus expression vector system (bevs). the level of expression is approximately 100-fold greater than in cem-c7 cells. between 0.5-1.0 mg hgr can be generated per liter of sf9 cell culture. the expressed hgr is capable of binding glucocorticoids with specificity and high affinity. covalent labeling with 3h-dexamethasone mesylate and western blot analysis using a polyclonal ant ...	1990	2330001
role of glycosylation in the transport of recombinant glycoproteins through the secretory pathway of lepidopteran insect cells.	cell lines established from the lepidopteran insect spodoptera frugiperda (e.g., sf9) are used routinely as hosts for the expression of foreign proteins by baculovirus vectors. previously, we showed that human tissue plasminogen activator (t-pa) was expressed, n-glycosylated, and secreted by sf9 cells infected with a recombinant baculovirus (jarvis dl, summers md: mol cell biol 9:214-223, 1989). we also showed that t-pa secretion was blocked by tunicamycin (tm), an inhibitor of n-glycosylation, ...	1990	2341487
cloning and expression of ndv hemagglutinin-neuraminidase cdna in a baculovirus expression vector system.	the hemagglutinin-neuraminidase (hn) gene of the hitchner b1 strain of newcastle disease virus (ndv) was cloned as a cdna and inserted into a baculovirus expression vector. the recombinant hn (rechn) expressed in spodoptera frugiperda cells had both hemagglutinating and neuraminidase activities both of which were inhibited by polyclonal anti-ndv sera or a monoclonal antibody (mab) against hn. infected insect cells could hemadsorb chicken red blood cells suggesting that the rechn is properly glyc ...	1990	2345960
polybrene-mediated transfection of cultured lepidopteran cells: induction of a drosophila heat shock promoter.	we have introduced hsp-cat plasmid dna into spodoptera frugiperda (lepidoptera: noctuidae) cells by transfection with purified dna (1 to 48 micrograms/ml) mixed with the polycation polybrene (100 micrograms/ml) in serum-free grace's medium. the hsp-cat construct contains a gene coding for the bacterial enzyme chloramphenicol acetyltransferase (cat), whose expression is controlled by a promoter derived from a drosophila heat shock (hsp) gene. expression of cat activity in transfected spodoptera c ...	1990	2384451
oligosaccharide processing in the expression of human plasminogen cdna by lepidopteran insect (spodoptera frugiperda) cells.	a comparison has been made between the asn289-linked oligosaccharide structures of human plasma plasminogen and a recombinant human plasminogen, expressed in lepidopteran insect (spodoptera frugiperda) cells, after infection of these cells with a recombinant baculovirus containing the entire human plasminogen cdna. using anion-exchange liquid chromatography mapping of the oligosaccharide units cleaved from the proteins by glycopeptidase f, compared with elution positions of standard oligosacchar ...	1990	2386787
identification and characterization of the gene encoding the major structural protein of insect iridescent virus type 22.	the major structural protein (msp--apparent molecular weight 49,000) of insect iridescent virus type 22 (isolated from blackflies--simulium spp.) was resolved from disrupted, purified virus particles by sds-page and transferred to nitrocellulose by western blotting. the portion of the blot containing the msp was identified and excised. tryptic peptides, generated by digestion in situ, were purified by hplc. three of these peptides were sequenced and an oligonucleotide gene probe was designed usi ...	1990	2389558
comparative study of virion structure, protein composition and genomic dna of three ascovirus isolates.	the virions of three ascoviruses isolated from the noctuids heliothis zea, spodoptera frugiperda and trichoplusia ni were compared with respect to their size and structure, protein composition and the size and relatedness of their dnas. the virions of the isolates from h. zea (hav) and t. ni (tav) were allantoid in shape (400 x 130 nm), enveloped and contained an inner particle which appeared to have an internal lipid bilayer surrounding the dna core. the virions of the s. frugiperda isolate (sa ...	1990	2391498
expression of the glycoprotein gene from a fish rhabdovirus by using baculovirus vectors.	a cdna fragment containing the gene encoding the glycoprotein of infectious hematopoietic necrosis virus was inserted into autographa californica baculovirus vectors under the control of the polyhedrin promoter. a 66-kilodalton protein, identical in size to the glycosylated glycoprotein of infectious hematopoietic necrosis virus, was expressed at high levels in spodoptera frugiperda cells infected with the recombinant viruses. the expressed protein reacted with antiserum to the glycoprotein on w ...	1990	2403442
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
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
the oligosaccharides of influenza virus hemagglutinin expressed in insect cells by a baculovirus vector.	the hemagglutinin of fowl plague virus has been expressed in spodoptera frugiperda (sf) cell cultures using a baculovirus vector. to elucidate the structure of the carbohydrate side chains, radioactively labeled oligosaccharides were liberated by treatment with endoglucosaminidase h and glycopeptidase f. sequential degradation with exoglycosidases and chromatographic analyses revealed the presence of oligomannosidic side chains, predominantly of the structures man5-9glcnac2, and the truncated ol ...	1990	2407026
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
synthesis, purification, and characterization of the cytoplasmic domain of the human insulin receptor using a baculovirus expression system.	the cytoplasmic domain of the beta subunit of the human insulin receptor has been overexpressed in insect cells using the baculovirus expression system. a recombinant baculovirus (bir-2) was constructed by inserting the human insulin proreceptor cdna fragment that encodes the cytoplasmic domain of the receptor into the genome of autographa californica nuclear polyhedrosis virus adjacent to the strong polyhedrin promoter. synthesis of the protein (baculovirus insulin receptor kinase (birk), mr 48 ...	1988	2451671
regulation of host rna levels during baculovirus infection.	during infection of the permissive host insect cell line spodoptera frugiperda iplb-sf-21 by the baculovirus autographa californica nuclear polyhedrosis virus (acmnpv), the levels of host actin, histone, and heat shock 70 (hsp70) rnas are reduced substantially. reduction of the host rna levels occurs primarily during a narrow window of the replication process, from approximately 12 to 18 hr postinfection (p.i.), corresponding to the phase in which the extracellular form of the virus buds into th ...	1988	2459844
vp4 protein of porcine rotavirus strain osu expressed by a baculovirus recombinant induces neutralizing antibodies.	the complete vp4 gene of porcine rotavirus strain osu has been inserted into a baculovirus expression vector under the control of the polyhedrin promoter. the vp4 outer capsid protein, which is a major neutralization antigen in rotavirus, was expressed in high yield in spodoptera frugiperda cells. reactivity with polyclonal and monoclonal antibodies suggested that neutralizing epitopes were functionally unaltered on the expressed vp4. the vp4 produced in this system also induced antibodies in gu ...	1989	2480689
glycosylation and secretion of human tissue plasminogen activator in recombinant baculovirus-infected insect cells.	cell lines established from the lepidopteran insect spodoptera frugiperda (fall armyworm; sf9) are used routinely as hosts for the expression of foreign proteins by recombinant baculovirus vectors. we have examined the pathway of protein glycosylation and secretion in these cells, using human tissue plasminogen activator (t-pa) as a model. t-pa expressed in sf9 cells was both n glycosylated and secreted. at least a subset of the n-linked oligosaccharides in extracellular t-pa was resistant to en ...	1989	2494430
high level expression of nonfused foreign genes with autographa californica nuclear polyhedrosis virus expression vectors.	high levels of nonfused chloramphenicol acetyltransferase, beta-galactosidase, and beta-glucuronidase expressed under the control of new vector constructs of the polyhedrin promoter in spodoptera frugiperda cells infected with autographa californica nuclear polyhedrosis virus were investigated by sds-page and rna dot blot analysis of total cytoplasmic rna. when the polyhedrin atg start codon was converted to att by site-directed mutagenesis, translation initiated at downstream atg codons resulti ...	1989	2497580
expression of functional human acid beta-glucosidase in cos-1 and spodoptera frugiperda cells.	a cdna encoding human acid beta-glucosidase (n-acylsphingosyl-1-o-beta-d-glucoside: glucohydrolase, ec 3.2.1.45) expressed catalytically active enzyme in transfected cos-1 or infected spodoptera frugiperda (sf9) cells. the expression plasmid p91023(b) (p91023b/glc) and a baculovirus (acmnpv/glc) containing the cdna were constructed and used with the respective cells. by immunoblotting a glycosylated, 63-kilodalton human acid-beta-glucosidase was detected in the transfected or infected cells. a 5 ...	1989	2498077
a baculovirus blocks insect molting by producing ecdysteroid udp-glucosyl transferase.	the predicted amino acid sequence of a newly identified gene of the insect baculovirus autographa californica nuclear polyhedrosis virus was similar to several uridine 5'-diphosphate (udp)-glucuronosyl transferases and at least one udp-glucosyl transferase. genetic and biochemical studies confirmed that this gene encodes an ecdysteroid udp-glucosyl transferase (egt). this enzyme catalyzes the transfer of glucose from udp-glucose to ecdysteroids, which are insect molting hormones. expression of t ...	1989	2505387
the rhesus rotavirus outer capsid protein vp4 functions as a hemagglutinin and is antigenically conserved when expressed by a baculovirus recombinant.	rhesus rotavirus (rrv) gene 4 was cloned into lambda bacteriophage, inserted into a polyhedrin promoter shuttle plasmid, and expressed in sf9 cells by a recombinant baculovirus. the baculovirus-expressed vp4 protein made up approximately 5% of the spodoptera frugiperda-infected cell protein. monoclonal antibodies that neutralize the virus bound to the expressed vp4 polypeptide, indicating that the expressed vp4 protein was antigenically indistinguishable from viral vp4. in addition, we have dete ...	1989	2538649
expression and purification of biologically active v-sis/platelet-derived growth factor b protein by using a baculovirus vector system.	malignant transformation induced by simian sarcoma virus is mediated by its v-sis protein, the monkey homolog of the platelet-derived growth factor (pdgf) b chain. by use of an appropriately engineered baculovirus expression vector, the v-sis protein was expressed in the insect cell line spodoptera frugiperda (sf9) at a level 50- to 100-fold higher than that observed with overexpression in mammalian-cell transfectants. the sis protein produced by sf9 cells underwent processing similar to that ob ...	1989	2542616
synthesis of immunogenic, but non-infectious, poliovirus particles in insect cells by a baculovirus expression vector.	a baculovirus expression vector (acleon) derived from autographa californica nuclear polyhedrosis virus (acnpv) was prepared containing the complete 6.6 kb coding region of the p3/leon/37 strain of poliovirus type 3 placed under the control of the acnpv polyhedrin promoter. the recombinant virus was used to infect spodoptera frugiperda insect cells. as demonstrated by use of the appropriate antibodies, infected insect cells made poliovirus proteins that included the structural proteins vp0, vp1 ...	1989	2543785
expression of two related nonstructural proteins of bluetongue virus (btv) type 10 in insect cells by a recombinant baculovirus: production of polyclonal ascitic fluid and characterization of the gene product in btv-infected bhk cells.	in vitro translation of bluetongue virus (btv) double-stranded rna in the rabbit reticulocyte lysate system has shown segment 10 (s10) to code for two related proteins, ns3 and ns3a. the presence of both products in vivo, however, has remained unconfirmed owing to the very low level of synthesis of the s10 gene product(s) in btv-infected bhk cells. in the present work, a cdna copy of btv type 10 (btv-10) s10 rna was inserted into autographa californica nuclear polyhedrosis baculovirus (acnpv) in ...	1989	2545903
rotavirus sa11 genome segment 11 protein is a nonstructural phosphoprotein.	we investigated properties of the rotavirus genome segment 11 protein. a rotavirus sa11 genome segment 11 cdna which contains the entire coding region was sequenced and inserted into the baculovirus transfer vector pvl941. recombinants containing gene 11 cdna were selected, and the gene 11 product expressed in spodoptera frugiperda cells infected with these recombinants was inoculated into guinea pigs to produce hyperimmune antiserum. characterization of the antiserum showed that it recognized a ...	1989	2548010
high-efficiency expression of mammalian beta-adrenergic receptors in baculovirus-infected insect cells.	infection of a clonal isolate of spodoptera frugiperda cells (sf9) with a baculovirus expression vector harboring the cdna encoding the beta-adrenergic receptor resulted in a high efficiency expression. at 48 hr post-infection, the level of expression of beta-adrenergic receptors was approximately 12 million/cell. specific activities of crude lysates of infected sf9 cells were approximately 30 pmol/mg of protein, 5-fold greater than those of membranes of high-expressor chinese hamster ovary cell ...	1989	2551283
expression of largest rna segment and synthesis of vp1 protein of bluetongue virus in insect cells by recombinant baculovirus: association of vp1 protein with rna polymerase activity.	the bluetongue virus core particles have been shown to contain an rna-directed rna polymerase (1). to identify the protein responsible for the virion rna polymerase activity, the complete 3.9 kb dna clone representing the largest rna segment 1 (l1) of bluetongue virus (btv-10) was placed under control of the polyhedrin promoter of autographa californica nuclear polyhedrosis virus (acnpv). the derived recombinant virus was used to infect spodoptera frugiperda cells. as demonstrated by stained pol ...	1989	2552409
synthesis of functional bovine leukemia virus (blv) p34tax protein by recombinant baculoviruses.	the bovine leukemia virus (blv) p34tax (also called tat, p34, xlor gene product) is a 34-kda polypeptide encoded in the 3'-terminal region of the virus. this protein is responsible for positive transcriptional trans-activation of promoter elements located within the blv long-terminal repeat. we introduced the protein-coding region of blv p34tax into the genome of the baculovirus autographa californica nuclear polyhedrosis virus. after infection of the insect spodoptera frugiperda (sf9) cell line ...	1989	2554576
overexpression and assembly of the herpes simplex virus type 1 helicase-primase in insect cells.	herpes simplex virus type 1 (hsv-1) encodes a helicase-primase that consists of three polypeptides encoded by the ul5, ul8, and ul52 genes (crute, j.j., tsurumi, t., zhu, l., weller, s.k., olivo, p.d., challberg, m.d., mocarski, e.s., and lehman, i.r. (1989) proc. natl. acad, sci, u.s.a. 86, 2186-2189). to obtain sufficient quantities of the enzyme for study, we have overexpressed the three genes using the baculovirus expression system. we find that the fully active enzyme can be assembled in vi ...	1989	2556383
expression of hemolytically active human complement component c1r proenzyme in insect cells using a baculovirus vector.	the gene of human c1r has been expressed in a baculovirus-insect-cell system via the pac373 transplacement vector. the full-length cdna copy was inserted into the pac373 vector downstream from the strong polyhedrin promoter of the baculovirus, autographa californica nuclear polyhedrosis virus (acnpv). spodoptera frugiperda cells were cotransfected with the resultant plasmid, pacc1r, and the wild-type acnpv dna. recombinant viruses, which drove the expression of c1r protein, were selected by plaq ...	1989	2557186
expression of sindbis virus 26s cdna in spodoptera frugiperda (sf9) cells, using a baculovirus expression vector.	to study protein processing in an insect spodoptera frugiperda (fall armyworm; sf9) cell line, a 26s cdna encoding the sequence of sindbis virus structural proteins (capsid protein, of 30 kilodaltons [kda]; p62 [the precursor of e3 and e2], of 62 kda; a 6-kda peptide; and the e1 protein, of 56 kda) was inserted into the genome of autographa californica nuclear polyhedrosis virus (acnpv) adjacent to the polyhedrin promoter. by immunoblot analysis with antisera directed against whole sindbis virus ...	1989	2644447
location and nucleotide sequence of the 25k protein missing from baculovirus few polyhedra (fp) mutants.	wild-type and few polyhedra (fp) mutants of the baculovirus autographa californica nuclear polyhedrosis virus (acmnpv) were studied to identify and sequence the gene encoding the 25-kda (25k) protein normally present in acmnpv-infected spodoptera frugiperda cells but which is often missing from fp mutant-infected cells. our previous study had mapped two overlapping late transcripts to the insertion site of host cell dna within the hindiii-i fragment (33.8 to 37.7 map units) of wild-type acmnpv. ...	1989	2644735
synthesis of biologically active influenza virus hemagglutinin in insect larvae.	the hemagglutinin of influenza (fowl plague) virus was expressed in larvae of heliothis virescens by using recombinant autographa californica nuclear polyhedrosis virus (acnpv) as a vector. animals were infected with the recombinant virus either by parenteral injection or by feeding. for oral uptake, recombinant virus occluded in polyhedra obtained from cultured spodoptera frugiperda cells after coinfection with authentic acnpv was used. immunohistological analyses of infected animals revealed t ...	1989	2648023
native and mutant 5-lipoxygenase expression in a baculovirus/insect cell system.	human 5-lipoxygenase (ec 1.13.11.34), the key enzyme involved in the transformation of arachidonic acid to the potent biologically active leukotrienes, has been overexpressed in insect cells using a baculovirus expression system. a recombinant baculovirus (3b6) carrying the human 5-lipoxygenase coding sequence downstream of the strong polyhedrin protein promoter was isolated. approximately 48 hr after infection of spodoptera frugiperda cells with the recombinant baculovirus, maximal intracellula ...	1989	2649885
glycosylation is not required for the fusion activity of the g protein of vesicular stomatitis virus in insect cells.	the gene encoding the complete glycoprotein of vesicular stomatitis virus (vsv, indiana serotype g protein) with potential asparagine-linked glycans at amino acid residues 179 and 338 was inserted into a baculovirus transfer vector pacym1, derived from the nuclear polyhedrosis virus of autographa californica (acnpv). the gene was placed under the control of the acnpv polyhedrin promotor and expressed by the derived recombinant viruses to high levels in spodoptera frugiperda cell lines. the princ ...	1989	2650461
insertion of the sfmnpv polyhedrin gene into an acmnpv polyhedrin deletion mutant during viral infection.	the autographa californica nuclear polyhedrosis virus (acmnpv) polyhedrin deletion mutant, d10a, was cotransfected with the recombinant plasmid sfmnpv.hindiii-l, which contains the spodoptera frugiperda nuclear polyhedrosis virus (sfmnpv) polyhedrin gene. an occlusion-positive hybrid virus was obtained which contained a dna insertion at approximately 13 map units on the acmnpv.d10a genome. a fine structure restriction map of cloned viral dna fragments from this region revealed that most of the 1 ...	1989	2655273
effect of cytochalasin d on cell morphology and acmnpv replication in a spodoptera frugiperda cell line.	cytochalasin d (cd) is a specific inhibitor of actin microfilament elongation and has been used to identify actin-dependent cellular processes. in this study we observed the effects of this inhibitor on autographa californica m nuclear polyhedrosis virus infected and uninfected iplb-sf-21 cells by electron microscopy. the cytochalasin d-induced morphological effects detected in uninfected cells included lobulate nuclei, double nuclei, long retraction processes, increased zeiosis, more frequent p ...	1989	2656869
expression of human plasminogen cdna in a baculovirus vector-infected insect cell system.	a cdna that encodes the human plasminogen (hpg) amino acid sequence has been inserted adjacent to the polyhedrin promoter in the genome of the baculovirus, autographa californica nuclear polyhedrosis virus, which was then used to infect cultured cells of the farm armyworm, spodoptera frugiperda. under the conditions of cell growth employed, recombinant (rec)-hpg was secreted into the medium after 24 h postinfection (p.i.), at which point virtually no rec-hpg antigen remained inside the cells. at ...	1989	2658810
analysis of transcription initiation in the panolis flammea nuclear polyhedrosis virus polyhedrin gene.	the nucleotide sequence of the polyhedrin gene of panolis flammea multiple nucleocapsid polyhedrosis virus (pfmnpv) has been determined. the coding sequences of this gene shared 82% similarity at the dna level and 88% similarity at the protein level with the polyhedrin gene from autographa californica (ac) mnpv. a single nucleotide deviation from the consensus transcription initiation sequence for baculovirus very late genes was identified in the pfmnpv polyhedrin gene. rna was prepared from mam ...	1989	2659729
studies on the control region of the p10 gene of the autographa californica nuclear polyhedrosis virus.	5' deletion mutants of the autographa californica nuclear polyhedrosis virus very late p10 gene promoter have been prepared and subjected to a transient expression assay in infected spodoptera frugiperda cells. the control plasmid contained the chloramphenicol acetyltransferase (cat) reporter gene under the control of the p10 promoter, which was included in a 230 bp sequence upstream from the p10 translation initiation codon. the control plasmid also contained a segment of the hr5 enhancer downs ...	1989	2659731
activation of the purified protein tyrosine kinase domain of the epidermal growth factor receptor.	biological responses to epidermal growth factor (egf) depend on the ligand-stimulated protein tyrosine kinase activity of its receptor. to further characterize the enzymatic activity of the egf receptor, the baculovirus expression system was used to express the cytoplasmic protein tyrosine kinase domain of the egf receptor. spodoptera frugiperda (sf9) cells infected with recombinant baculovirus correctly expressed an active tyrosine kinase domain of the egf receptor as demonstrated by 35s metabo ...	1989	2661557
expression of yb1 glutathione s-transferase using a baculovirus expression system.	a full-length cdna clone was isolated for rat liver yb1 glutathione s-transferase (ec 2.5.1.18). the coding sequence of yb1 cdna was inserted into a baculovirus vector for infection of spodoptera frugiperda (sf9) cells. the enzymatically active recombinant yb1 glutathione s-transferase protein has a native molecular weight of 42,000 daltons (by molecular sieve chromatography), a subunit molecular weight of 26,500 daltons (by sds-polyacrylamide gel electrophoresis), a pi of 8.4 and an extinction ...	1989	2669745
baculovirus polyhedrin promoter-directed expression of rubella virus envelope glycoproteins, e1 and e2, in spodoptera frugiperda cells.	to study the capability of spodoptera frugiperda (fall armyworm; sf9) cells to synthesize and process mature rubella virus (rv) proteins, a cdna encoding the structural envelope glycoproteins, e1 (58 kda) and e2 (42-47 kda) were inserted into the genome of autographa californica nuclear polyhedrosis virus (acnpv) and expressed during infection under the transcriptional regulation of the polyhedrin gene promoter. by immunoblot analysis with antibodies directed against purified rv, the individual ...	1989	2672567
novel exogenous heme-dependent expression of mammalian cytochrome p450 using baculovirus.	rat p450 iia1 is a membrane-bound hemoprotein that catalyzes the 7 alpha- and 6 alpha-hydroxylation of testosterone. a recombinant baculovirus, containing the cdna encoding iia1, was constructed and used to infect spodoptera frugiperda cells. infected cells contained up to 10% iia1 protein as quantified by western blot analysis; however, only a small percentage of this protein contained heme and was catalytically active. addition of hemin to the culture medium during the course of viral infectio ...	1989	2679389
expression and characterization of the ha-ras p21 protein produced at high levels in the insect/baculovirus system.	normal and mutated cdnas of ha-ras have each been cloned into a standard (pac373) and a novel (p36c) baculovirus transfer vector and introduced via homologous recombination into the genome of autographa californica nuclear polyhedrosis virus immediately downstream of the polyhedrin promoter. spodoptera frugiperda cells infected with recombinant virus containing the normal ha-ras gene express very high levels of ras p21 protein (approximately 20% of total cell protein), whereas the mutant protein ...	1989	2681210
construction and analysis of an autographa californica nuclear polyhedrosis virus mutant lacking the polyhedral envelope.	an autographa californica nuclear polyhedrosis virus (acmnpv)-specific protein of 34 kda (pp34) is shown to be involved in the morphogenesis of the polyhderal envelope of baculoviruses. the region of the acmnpv genome encompassing ecori-h and -s (map positions 82.6-85.8) contains five open reading frames (orfs) forming one transcriptional unit. the bacterial beta-galactosidase (lacz) gene was inserted in phase with the n-terminal 12 amino acids of orf3, thereby intervening this gene. a recombina ...	1989	2683364
immunogenic and protective properties of rabies virus glycoprotein expressed by baculovirus vectors.	the gene encoding the glycoprotein of rabies virus (g protein, cvs strain) has been cloned and inserted into the baculovirus transfer vector pacym1 derived from the nuclear polyhedrosis virus of autographa californica (acnpv). the gene was placed under the control of the acnpv polyhedrin promoter and expressed to high levels by the derived recombinant virus using a spodoptera frugiperda cell line. it has been established that the antigenic characteristics of the protein were conserved by compari ...	1989	2688298
characterization of japanese encephalitis virus envelope protein expressed by recombinant baculoviruses.	recombinant baculoviruses containing the coding sequences of the viral structural proteins, i.e., the capsid (c) protein, the precursor to premembrane (prem) protein, and the envelope (e) protein, as well as a nonstructural protein, ns1, of japanese encephalitis virus (jev) were constructed. infection of spodoptera frugiperda cells with these recombinant viruses produced prem and e proteins. the e proteins synthesized by the recombinants were shown to be glycosylated and similar in size to the a ...	1989	2688300
efficient transfection of insect cells with baculovirus dna using electroporation.	efficient transfection of spodoptera frugiperda cells with autographa californica nuclear polyhedrosis virus (acnpv) dna has been carried out using the technique of electroporation. the efficiency of transfection was monitored by assaying the extracellular virus in cell culture supernatants 3 days post-electroporation. maximum titres of 2 x 10(9) p.f.u./ml acnpv were obtained when using a pulse length of 7.7 ms and a field strength of 500 v/cm. this compared with a titre of 2 x 10(6) p.f.u./ml a ...	1989	2691634
high-level expression and purification of a recombinant human erythropoietin produced using a baculovirus vector.	conditions presently have been established for the high-level expression and simplified purification of recombinant human erythropoietin produced in spodoptera frugiperda cells. expression, as mediated by infection with a recombinant baculovirus, was accomplished in suspension culture using reduced levels of serum and media supplements experimentally determined to provide optimum levels of factor production (500,000 u/l). purification of this recombinant human erythropoietin to virtual homogenei ...	1989	2752139
insect cells as substrates for biologicals.	various methods for the growth of insect cells and the production of recombinant human beta-interferon (rhu beta-ifn) by insects cells, spodoptera frugiperda, ipl-sf-21ae after infection with recombinant acnpv are described. using a suspension perfusion biospin filter culture system, an ipl-sf-21ae cell line was propagated in a semi-controlled environment. to obtain and maintain high cell densities and viabilities in this system, the culture medium for suspension methods was modified by suppleme ...	1989	2759354
[use of baculovector for the expression of hbsag gene in insect cells].	based on the information of molecular biology of autographa californica nuclear polyhedrosis virus (acnpv), a recombinant transfer plasmid pacmv was constructed by molecular procedures included using two synthetic localized probes, which provided an inserted position linked with bamhi sequences nearly at polyhedrin initiating atg codon. then an expression vector pacmv-hbsag was reconstructed, it contained hbsag gene from subclone pypss-1 derived from adwserotype of hbv. the recombinant virus con ...	1989	2800542
cell-free translation in lysates from spodoptera frugiperda (lepidoptera: noctuidae) cells.	1. conditions for in vitro translation of mrna in cell-free extracts from cultured spodoptera frugiperda cells were defined. 2. incorporation of [35s]methionine into acid-precipitable material increased for approximately 1 hr, and was sensitive to the protein synthesis inhibitors pactamycin and cycloheximide. 3. micrococcal nuclease-treated lysate, primed with purified rabbit globin mrna, synthesized a major protein with the size of full length globin, indicating that the lysate supported correc ...	1989	2805641
identification and nucleotide sequence of the regions of autographa californica nuclear polyhedrosis virus genome carrying insertion elements derived from spodoptera frugiperda.	a morphology mutant of the baculovirus autographa californica nuclear polyhedrosis virus called m5 was previously shown to synthesize two size classes of viral dna, one of which had a deletion of 42% of the genome. it was hypothesized that the presence of the smaller m5 circular dna resulted from the specific deletion of the region located between two sites carrying short dna insertions. these sites have now been identified by dna-dna hybridization using cloned ecori fragments of m5 dna. one clo ...	1987	2823472
functional mapping of autographa california nuclear polyhedrosis virus genes required for late gene expression.	a plasmid containing the bacterial chloramphenicol acetyltransferase (cat) gene under the control of an autographa california nuclear polyhedrosis virus (acnpv) late gene promoter was constructed. this plasmid (pl2cat) also contained the acnpv hr5 enhancer element. transient-expression assay experiments indicated that the late promoter was active in spodoptera frugiperda cells cotransfected with pl2cat and acnpv dna but not when pl2cat was transfected alone. low levels of cat activity were obser ...	1988	2826808
comparisons of host cell dna insertions and altered transcription at the site of insertions in few polyhedra bacilovirus mutants.	few polyhedgra (fp) mutants of autographa californica nuclear polyhedrosis virus (acnpv) and the closely related strain galleria mellonella (gm)npv have been reported which contain trichoplusia ni host cell dna sequences inserted into the viral genome between map units 35.0 and 37.7. new fp mutants are described with alterations of the hindiii-i restriction enzyme fragment (33.8 to 37.7 map units) of acnpv, either deletions of viral dna sequences or insertions of spodoptera frugiperda host cell ...	1988	2829419
efficient expression in insect cells of a soluble, active human insulin receptor protein-tyrosine kinase domain by use of a baculovirus vector.	the human insulin receptor (ir) is a transmembrane glycoprotein, whose cytoplasmic domain contains an insulin-activated protein-tyrosine kinase (ec 2.7.1.112). by the use of an appropriately engineered baculovirus expression vector, a soluble cytoplasmic derivative of this domain was expressed in the insect cell line spodoptera frugiperda (sf9). at 24 to 48 h after sf9 cells were infected with recombinant virus, a protein of the size expected for this domain (approximately 48 kilodaltons) consti ...	1988	2833613