Publications

use of recombinant baculoviruses in synthesis of morphologically distinct viruslike particles of flock house virus, a nodavirus.	flock house virus (fhv) is a small icosahedral insect virus of the family nodaviridae. its genome consists of two messenger-sense rna molecules, both of which are encapsidated in the same particle. rna1 (3.1 kb) encodes proteins required for viral rna replication; rna2 (1.4 kb) encodes protein alpha (43 kda), the precursor of the coat protein. when spodoptera frugiperda cells were infected with a recombinant baculovirus containing a cdna copy of rna2, coat protein alpha assembled into viruslike ...	1993	8474173
functional expression of the rat anion exchanger ae2 in insect cells by a recombinant baculovirus.	we used baculovirus to transiently express a rat anion exchanger (ae2) in spodoptera frugiperda (sf9) insect cells. no detectable cl(-)-hco3- exchange activity was observed in wild type or sham-infected sf9 cells, monitored using 2',7'-bis(2-carboxyethyl)-5(6)-carboxyfluorescein, a ph-sensitive fluorescent dye. functional expression of anion exchange activity in the ae2 recombinant baculovirus-infected cells was observed within the first day after infection and sustained over the next 3 days. th ...	1993	8476013
the 35-kilodalton protein gene (p35) of autographa californica nuclear polyhedrosis virus and the neomycin resistance gene provide dominant selection of recombinant baculoviruses.	autographa californica nuclear polyhedrosis virus (acmnpv) recombinants were constructed to test the effectiveness of the acmnpv 35-kilodalton protein gene (35k gene) and the bacterial neomycin resistance gene (neo) as dominant selectable markers for baculoviruses. insertion of the acmnpv apoptosis suppressor gene (p35) into the genome of p35-deletion mutants inhibited premature host cell death and increased virus yields up to 1200-fold at low multiplicities in spodoptera frugiperda (sf21) cell ...	1993	8493093
high level expression of the large subunit of mouse ribonucleotide reductase in a baculovirus system.	the large subunit of ribonucleotide reductase from mouse has been overexpressed in spodoptera frugiperda cells infected with recombinant baculovirus. the expressed protein was purified by affinity chromatography to apparent homogeneity as determined by sds-page. the homogeneous protein is recognized in western blot analysis by a monoclonal antibody raised to the large subunit of ribonucleotide reductase from calf thymus, has the correct n-terminal sequence, and, in the presence of the small subu ...	1993	8495754
identification and characterization of lef-1, a baculovirus gene involved in late and very late gene expression.	an autographa californica nuclear polyhedrosis virus (acmnpv) gene required in transient expression assays for late and very late viral gene expression was identified, sequenced, and transcriptionally mapped. this gene, designated late expression factor 1 (lef-1), was located between 7.4 and 8.7 map units of the acmnpv physical map. it was identified by cotransfecting spodoptera frugiperda cultured cells with a collection of overlapping cloned dna fragments covering the entire acmnpv genome and ...	1993	8497062
post-translational processing and thr-206 are required for glycosylasparaginase activity.	lysosomal glycosylasparaginase is encoded as a 36.5 kda polypeptide that is post-translationally processed to subunits of 19.5 kda (heavy) and 15 kda (light). recombinant glycosylasparaginase has been expressed in spodoptera frugiperda insect cells enabling the precursor and processed forms to be isolated and their catalytic potential determined. only the subunit conformation was functional indicating glycosylasparaginase is encoded as an inactive zymogen. the newly created amino terminal residu ...	1993	8500622
isolation and expression of human 1,25-dihydroxyvitamin d3 24-hydroxylase cdna.	human 1,25-dihydroxyvitamin d3 24-hydroxylase cdna clones were isolated from an hl-60 cell cdna library by using a reverse transcription/polymerase chain reaction-generated human cdna probe. the 24-hydroxylase cdna consists of a 1539-bp open reading frame encoding a 513-amino acid polypeptide. protein sequence analysis shows that the human 24-hydroxylase is 90% homologous (82% identical) to that of the rat, with 100% homology in the 21-amino acid heme-binding region. northern blot analysis showe ...	1993	8506296
breaching the conformational integrity of the catalytic triad of the serine protease plasmin: localized disruption of a side chain of his-603 strongly inhibits the amidolytic activity of human plasmin.	site-directed mutagenesis has been used to construct a cdna that encodes a recombinant variant human plasminogen (hpg) containing a pro-611-->ile mutation (mrhpg). the mutein was expressed in recombinant baculovirus-infected spodoptera frugiperda cells (iplb-sf-21ae), and purified. after activation of this zymogen to its corresponding form of the serine protease plasmin (mrhpm), this latter enzyme was essentially inactive toward an amide plasmin substrate, most likely from alteration of the spat ...	1993	8506386
apoptosis reduces both the in vitro replication and the in vivo infectivity of a baculovirus.	apoptotic programmed cell death occurs when the insect cell line sf-21, derived from spodoptera frugiperda, is infected with mutants of the baculovirus autographa californica nuclear polyhedrosis virus (acmnpv) which lack a functional p35 gene. however, infection of the trichoplusia ni tn-368 cell line with p35 mutants does not result in apoptosis (r. clem, m. fechheimer, and l. miller, science 254:1388-1390, 1991). we have examined the effect of apoptosis on acmnpv infections in cell lines and ...	1993	8510202
high-level production of biologically active human cytosolic phospholipase a2 in baculovirus-infected insect cells.	infection of spodoptera frugiperda insect cells with a recombinant baculovirus expressing human cytosolic phospholipase a2 (cpla2) resulted in the production of biologically active protein. the level of recombinant human cpla2 production in infected insect cells was at least 50-fold higher than that observed in human monoblast u937 cells.	1993	8514199
location of two putative origins of dna replication of autographa californica nuclear polyhedrosis virus.	previously, we described a defective autographa californica nuclear polyhedrosis virus (acmnpv) which must contain cis-acting elements required for dna synthesis, such as the origin(s) of replication (ori). defective genomes of acmnpv generated after serial undiluted passage were analyzed further. three small separated regions were retained in dna of defective acmnpv and accumulated in extracellular defective interfering viruses as well as in intracellular dna after 40 passages. two of these reg ...	1993	8517035
the 85-kda, arachidonic acid-specific phospholipase a2 is expressed as an activated phosphoprotein in sf9 cells.	a human, 85-kda arachidonoyl-specific, cytosolic phospholipase a2 was expressed using the baculovirus-insect cell expression system. expression resulted in the production of an active protein which consisted of approximately 3% of the total protein in the host spodoptera frugiperda (sf9) cells at 67 h after infection. the phospholipase a2 was purified to apparent homogeneity and exhibited calcium-dependent phospholipase a2 activity with a specific activity of 8 mumol/min/mg protein, as well as c ...	1993	8215460
human rotavirus k8 strain represents a new vp4 serotype.	the complete vp4 gene of the human rotavirus (hrv) k8 strain (g1 serotype) was cloned and inserted into the baculovirus transfer vector pvl941 under the control of the polyhedrin promoter. a k8vp4 recombinant baculovirus was obtained by cotransfection of spodoptera frugiperda (sf9) cells with transfer vector dna containing the k8vp4 gene and wild-type baculovirus dna. infection of sf9 cells with this vp4 recombinant baculovirus resulted in the production of a protein that is similar in size and ...	1993	8380098
formation of poliovirus-like particles by recombinant baculoviruses expressing the individual vp0, vp3, and vp1 proteins by comparison to particles derived from the expressed poliovirus polyprotein.	the cdna sequences encoding the vp0, vp3, and vp1 structural proteins of poliovirus type 3 (strain p3/leon/37) have been individually cloned and used to prepare recombinant baculoviruses based on autographa californica nuclear polyhedrosis viruses (acnpv). expression of the proteins was monitored in virus-infected spodoptera frugiperda cells. both the individually expressed vp0 protein and vp0 derived from the complete poliovirus coding region were shown to incorporate myristic acid. a significa ...	1993	8380663
expression of rat ctp:phosphocholine cytidylyltransferase in insect cells using a baculovirus vector.	ctp:phosphocholine cytidylyltransferase (ct) is a key regulatory enzyme in phosphatidylcholine biosynthesis. we constructed a recombinant baculovirus (bct) containing rat ct cdna under the control of the polyhedrin promoter. crude cell extracts of spodoptera frugiperda (sf9) cells infected with bct possessed 250-fold higher specific activities for ct compared to rat liver cytosol, and ct protein constituted 3-6% of the total cellular protein. the 42-kda form of ct predicted from the cdna sequenc ...	1993	8382903
penetration of autographa californica nuclear polyhedrosis virus nucleocapsids into iplb sf 21 cells induces actin cable formation.	the budded form of autographa californica nuclear polyhedrosis virus functionally enters cells by adsorptive endocytosis. during the period of virus uptake into spodoptera frugiperda iplb sf21 cells (1 to 4 hr postinfection), filamentous f-actin cables, visualized by fluorescence microscopy, were formed within the cytoplasm of infected cells. cable formation appeared to be a direct effect of viral inoculum in that the numbers of observed cables increased with an increase in multiplicity of infec ...	1993	8212560
phosphorylation of synthetic peptides containing tyr-met-x-met motifs by nonreceptor tyrosine kinases in vitro.	several tyrosine phosphorylation sites in the insulin receptor kinase substrate irs-1 are predicted to be within tyr-met-x-met (ymxm) motifs, and synthetic peptides corresponding to these sequences are excellent substrates for the insulin receptor kinase in vitro (shoelson, s. e., chatterjee, s., chaudhuri, m., and white, m. f. (1992) proc. natl. acad. sci. u. s. a. 89, 2027-2031). in this study, ymxm-containing peptides are shown to act as substrates for two members of the nonreceptor subfamily ...	1993	8227078
the antimicrosporidial activity of albendazole.	the antimicrosporidial activity of albendazole was tested on nosema bombycis in vitro in spodoptera frugiperda cells and in vivo in heliocoverpa zea larvae and pupae. significant reductions in the percentage of infected s. frugiperda cells were obtained using a concentration of 5.3 micrograms/ml albendazole in tissue culture medium but recrudescence occurred after the drug was withdrawn from the cultures. significant reductions in the number of spores harvested from 6th-instar larvae or pupae we ...	1993	8228321
expression of two different forms of cdna for thromboxane synthase in insect cells and site-directed mutagenesis of a critical cysteine residue.	cdna coding for human placental thromboxane synthase (ec 5.3.99.5) was amplified by pcr from a human placental cdna library and sequenced. this cdna and a shorter cdna isolated from a human lung cdna library with a deletion of 163 bp near the 3' end were expressed in spodoptera frugiperda (sf9) insect cells using a baculovirus expression system. the cdna from human placenta was expressed as an active enzyme (60 kda) with a specific activity higher than those reported from other cell types, where ...	1993	8240243
construction of a cdna encoding the multifunctional animal fatty acid synthase and expression in spodoptera frugiperda cells using baculoviral vectors.	a cdna encoding the 2505-residue multifunctional rat fatty acid synthase has been constructed and expressed as a catalytically active protein in spodoptera frugiperda (sf9) cells using autographa californica nuclear polyhedrosis virus (baculovirus). the 7.5 kb cdna was engineered by the amplification and sequential splicing together of seven fragments contained in overlapping cdnas that collectively spanned the entire rat fatty acid synthase coding sequence. the full-length cdna was cloned into ...	1993	8250834
adenine dna methyltransferase m.cviri expression accelerates apoptosis in baculovirus-infected insect cells.	the adenine dna methyltransferase m.cviri (tgcma) gene from chlorella virus xz-6e was cloned into the autographa californica nuclear polyhedrosis virus (acmnpv) genome and expressed in spodoptera frugiperda insect cells under the control of two tandemly arranged viral promoters, the early etl promoter and the late polyhedrin promoter. m.cviri activity was first detected at 10 hr p.i and reached a maximum at 48 hr p.i. viral dna synthesized in insect cells infected with m.cviri expressing virus ( ...	1993	8372450
comparative recombinant protein production of eight insect cell lines.	a recombinant autographa californica baculovirus expressing secreted alkaline phosphatase (seap) gene was used to evaluate the expression of a secreted glycoprotein in eight insect cell lines derived from spodoptera frugiperda, trichoplusia ni, mamestra brassicae and estigmene acrea. because cell density was found to influence protein production, seap production was evaluated at optimal cell densities for each cell line on both a per cell and per milliliter basis. on a per cell basis, the t. ni- ...	1993	8314732
biosynthesis and secretion of human interleukin 2 glycoprotein variants from baculovirus-infected sf21 cells. characterization of polypeptides and posttranslational modifications.	human interleukin 2 (il-2) and human il-2 mutant proteins, with artificially introduced n-glycosylation or o-glycosylation sites, have been expressed in a lepidopteran cell line (sf21, spodoptera frugiperda) using recombinant baculovirus vectors. only approximately 25% of the total recombinant il-2 protein synthesized by sf21 cells was secreted into the culture medium. significant n-terminal truncations were detected in the secreted polypeptides (up to 85% of the molecules). alanine and proline ...	1993	8344280
inhibition of african swine fever virus binding and infectivity by purified recombinant virus attachment protein p12.	the african swine fever virus protein p12, involved in virus attachment to the host cell, has an apparent molecular mass of 17 kda in sodium dodecyl sulfate-polyacrylamide gel electrophoresis under nonreducing conditions. we have also identified 12- and 10-kda forms of the p12 protein in infected vero cells and found that the mature 17-kda protein is the only form present in virus particles. the p12 protein has been produced in large amounts in spodoptera frugiperda insect cells infected with a ...	1993	8350406
azadirachtin inhibits proliferation of sf 9 cells in monolayer culture.	azadirachtin a in ppm quantity inhibits proliferation and monolayer formation of spodoptera frugiperda (sf9) insect cells in monolayer culture. the incubated cells demonstrate reduced rates of protein synthesis which finally leads to cell death. this growth inhibiting activity is compared with other botanicals such as ohchinin, salannin and volkensin. the high and specific activity of azadirachtin a on insect cells is discussed in comparison with its effect on a mammalian cell line, based on 2 d ...	1993	8363710
expression and characterization of the rat d3 dopamine receptor: pharmacologic properties and development of antibodies.	a baculovirus expression system provided an enriched source of biologically and immunologically active d3 dopamine receptors. receptors expressed in spodoptera frugiperda insect (sf9) cells at a density of 5 to 15 pmol/mg of protein displayed high affinity for the antagonists, eticlopride, fluphenazine and spiroperidol, and the agonist, n-propylnorapomorphine. the binding of agonists was not sensitive to gtp. antisera raised against synthetic peptides in the third intracellular loop of the d3 do ...	1993	8437101
baculovirus-directed expression of human prostatic steroid 5 alpha-reductase 1 in an active form.	in the human prostate, the enzyme steroid 5 alpha-reductase (h5 alpha r) catalyses the conversion of testosterone into the more potent androgen, dihydrotestosterone. two distinct cdnas coding for h 5 alpha r in the human prostate have been previously characterized. enzyme h5 alpha r1 shows a maximum activity at basic ph whereas h5 alpha r2 has an acidic ph optimum activity. we report here the expression of the human steroid h5 alpha r1 in a eukaryotic expression system: the baculovirus-directed- ...	1993	8664165
semliki forest virus capsid protein expressed by a baculovirus recombinant.	we have constructed a recombinant baculovirus which expressed the semliki forest (sfv) capsid (c) gene as a fusion protein under the control of the polyhedrin gene promoter. the sequence coding for c and part of the envelope e3 region were expressed as a polyprotein precursor. spodoptera frugiperda (sf9) insect cells infected with the recombinant virus produced a protein reacting with polyclonal rabbit antiserum that had been raised against the 33 kda authentic c protein purified from sfv. prime ...	1993	8379852
functional expression of the alpha 2 and alpha 3 isoforms of the na,k-atpase in baculovirus-infected insect cells.	multiple isoforms of both the alpha and beta subunits of na,k-atpase have been identified. elucidating their roles has been complicated by the fact that most tissues express multiple isoforms and purification techniques specific for each isoform have not been achieved. the baculovirus expression system, which uses the baculovirus autographica californica to infect insect cells, is an ideal system for studying the na,k-atpase isoforms since high amounts of foreign proteins can be produced and som ...	1993	8383329
baculovirus expression of the maize mitochondrial protein urf13 confers insecticidal activity in cell cultures and larvae.	the urf13 protein, which is encoded by the mitochondrial gene t-urf13, is responsible for cytoplasmic male sterility and pathotoxin sensitivity in the texas male-sterile cytoplasm (cms-t) of maize. mitochondrial sensitivity to two host-specific fungal toxins (t toxins) is mediated by the interaction of urf13 and t toxins to form pores in the inner mitochondrial membrane. a carbamate insecticide, methomyl, mimics the effects of t toxins on isolated cms-t mitochondria. urf13 was expressed in spodo ...	1993	8475086
in vitro transcription from baculovirus late gene promoters: accurate mrna initiation by nuclear extracts prepared from infected spodoptera frugiperda cells.	extracts prepared from nuclei of autographa californica nuclear polyhedrosis virus-infected spodoptera frugiperda cells were shown to support in vitro transcription from baculovirus late gene promoters. in vitro transcription was optimized for the late promoter of the 39k gene. the mg2+ concentration was critical; concentrations higher than 1 to 2 mm did not support late transcription. additional conditions included template (40 micrograms/ml), extract (2.5 mg/ml), and incubation time (25 min). ...	1993	8510206
interaction of hepatic microsomal epoxide hydrolase derived from a recombinant baculovirus expression system with an azarene oxide and an aziridine substrate analogue.	a recombinant baculovirus (vehx) encoding rat hepatic microsomal epoxide hydrolase has been constructed. infection of spodoptera frugiperda (sf9) cells with the recombinant virus results in the expression of the enzyme at a level estimated to be between 5% and 10% of the cellular protein. the enzyme, which can be purified in 15% yield by a simple three-step procedure involving detergent extraction, deae-cellulose chromatography, and removal of the detergent on hydroxylapatite, has physical and k ...	1993	8383521
characterization of the human oxysterol receptor overexpressed in the baculovirus system.	oxysterols are potent regulators of enzymes of the de novo cholesterol biosynthetic pathway and do not require the ldl (low density lipoprotein):ldl receptor system for their regulatory actions. the search for an alternate transduction system led to the identification of an oxysterol binding protein. this cytosolic protein has been extensively characterized, purified, and cloned. although it fulfills the pharmacologic criteria for an oxysterol receptor by binding to oxysterols with affinities co ...	1993	8251955
an unusual 30-kda protein binding to the polyhedrin gene promoter of autographa californica nuclear polyhedrosis virus.	gel retardation assays using nuclear extracts from the autographa californica nuclear polyhedrosis virus-infected sf21 (spodoptera frugiperda) insect cell line revealed a host factor (polyhedrin promoter-binding protein or ppbp) that binds to the polyhedrin gene promoter. a hexanucleotide sequence (aataaa) within the promoter is important for binding in association with neighboring elements, some of which are contributed by the sequence taagtatt present at the transcription start point. ppbp was ...	1994	8300607
structures of the n-linked oligosaccharides of the membrane glycoproteins from three lepidopteran cell lines (sf-21, izd-mb-0503, bm-n).	the primary structures of the asn-linked carbohydrate chains isolated from membrane glycoproteins of the three insect cell lines mamestra brassicae (mb-0503), bombyx mori (bm-n), and spodoptera frugiperda (sf-21) have been determined. tryptic glycopeptides derived from the membrane fraction were digested with peptide-n-glycanase a. the resulting oligosaccharides were reductively aminated with 2-aminopyridine and identified by two-dimensional hplc mapping in combination with exoglycosidase digest ...	1994	8311447
a prototype recombinant vaccine against respiratory syncytial virus and parainfluenza virus type 3.	we have produced a genetically-engineered chimeric protein composed of the external domains of the respiratory syncytial virus (rsv) fusion (f) protein and the parainfluenza virus type 3 (piv-3) hemagglutinin-neuraminidase (hn) protein in insect cells using the baculovirus expression system. the yield of the soluble chimeric frsv-hnpiv-3 protein could be increased approximately 2-fold by using trichoplasia ni (high five) insect cells in place of spodoptera frugiperda (sf9) for expression. the ch ...	1994	7765021
analysis of human acid beta-glucosidase by site-directed mutagenesis and heterologous expression.	structure/function relationships of acid beta-glucosidase, the enzyme deficient in gaucher disease, were evaluated by characterizing the proteins expressed from cdnas encoding normal and mutant enzymes. twenty-two gaucher disease mutations or created mutations were expressed in spodoptera frugiperda (sf9) cells and analyzed for catalytic properties, stability, inhibitor binding, and modifier interactions. many gaucher disease mutations encoded highly disruptive amino acid substitutions (e.g. p28 ...	1994	8294487
expression and ligand binding characterization of the beta-subunit (p75) ectodomain of the interleukin-2 receptor.	the baculovirus-mediated eukaryotic insect cell expression system was used to prepare large quantities of the beta-subunit ectodomain of the high-affinity interleukin-2 receptor (il-2r beta x). we describe the expression, purification, and biophysical characterization of this ligand binding domain. the human cdna encoding il-2r beta x was inserted into baculovirus transfer vectors. high titer recombinant baculovirus was produced in spodoptera frugiperda (sf9) insect cells, and the viral supernat ...	1994	8180212
flow cytometric analysis of bioluminescence emitted by recombinant baculovirus-infected insect cells.	five recombinant baculoviruses, each containing a different insect luciferase gene encoding a protein with characteristic light emission properties, namely, luc gr (546 nm), luc ff (556 nm), luc yg (560 nm), luc ye (578 nm), and luc or (593 nm) were constructed. all genes were inserted under the transcriptional control of the polyhedrin gene promoter of the autographa californica nuclear polyhedrosis virus (acnpv) and expressed in spodoptera frugiperda insect cells during viral infection. the bi ...	1994	8187580
biologically active recombinant human complement factor h: synthesis and secretion by the baculovirus system.	a complete cdna clone (4.3 kb) encoding human complement factor h (hcfh; 155 kda) has been cloned into the pvl1393 baculovirus expression vector and transfected into spodoptera frugiperda sf9 insect cells. a biologically active (92 +/- 15%) 140-kda protein was secreted into the medium with a yield of more than 5 mg/liter. this is the first report of synthesis of biologically active recombinant hcfh in a heterologous system.	1994	8206393
crystallization and initial x-ray crystallographic characterization of recombinant bovine inositol polyphosphate 1-phosphatase produced in spodoptera frugiperda cells.	bovine inositol polyphosphate 1-phosphatase, a monomeric protein with a molecular mass of 44,000 da, hydrolyzes the 1-position phosphate from inositol 1,3,4-trisphosphate and inositol 1,4-bisphosphate. the low abundance of inositol polyphosphate 1-phosphatase in tissues has precluded structural studies requiring large quantities of enzyme. we used recombinant baculovirus harboring the cdna of bovine inositol polyphosphate 1-phosphatase to infect spodoptera frugiperda (sf9) insect cells. recombin ...	1994	8107142
synthesis of recombinant baculoviruses expressing the outer capsid protein vp2 of five btv serotypes and the induction of neutralizing antibodies to homologous and heterologous btv serotypes.	dna representing rna segment l2 of 5 different bluetongue virus (btv) serotypes (btv-1, -2, -11, -13 and -17) corresponding to the gene that codes for the btv neutralization antigen vp2, have been inserted individually into baculovirus transfer vectors in lieu of the 5' coding region of the polyhedrin gene of autographa california nuclear polyhedrosis virus (acnpv). after co-transfection of spodoptera frugiperda cells with wild-type acnpv dna in the presence of the derived transfer vector dnas, ...	1994	8178571
cellular toxicities and membrane binding characteristics of insecticidal crystal proteins from bacillus thuringiensis toward cultured insect cells.	the pattern of in vitro toxicity of activated toxins from several classes of entomocidal inclusion genes from bacillus thuringiensis was measured using eight established cell lines from lepidopteran insects. protoxins representing cryia(b), cryia(c), and a mixture of all three cryia toxins (subtypes a, b, and c; from b. thuringiensis subsp. kurstaki hd-1) were compared with the protoxin representing cryic in a bioassay which measured the viability of cultured insect cells upon exposure to entomo ...	1994	8176242
accurate processing and secretion in the baculovirus expression system of an erythroid-cell-stimulating factor consisting of a chimaera of insulin-like growth factor ii and an insect insulin-like peptide.	a synthetic gene encoding the signal peptide and the n-terminal sequence of bombyxin, an insect insulin-like peptide, and the 58 amino acids of the c-terminal sequence of human insulin-like growth factor ii (igf ii) has been expressed using the baculovirus system. this synthetic chimaera was obtained by amplification of four overlapping oligonucleotides using taq polymerase and cloning into the transfer vector pbluebac. the construct was integrated by homologous recombination into the autographa ...	1994	8166627
an apoptosis-inhibiting gene from a nuclear polyhedrosis virus encoding a polypeptide with cys/his sequence motifs.	two different baculovirus genes are known to be able to block apoptosis triggered upon infection of spodoptera frugiperda cells with p35 mutants of the insect baculovirus autographa californica nuclear polyhedrosis virus (acmnpv):p35 (p35-encoding gene) of acmnpv (r. j. clem, m. fechheimer, and l. k. miller, science 254:1388-1390, 1991) and iap (inhibitor of apoptosis gene) of cydia pomonella granulosis virus (cpgv) (n. e. crook, r. j. clem, and l. k. miller, j. virol. 67:2168-2174, 1993). using ...	1994	8139034
properties of a recombinant kainate receptor expressed in baculovirus-infected insect cells.	glur6 is a glutamate receptor of the kainate subtype that is expressed in the mammalian central nervous system. the cdna coding for the rat brain receptor was subcloned into a baculovirus expression vector and the purified recombinant virus was used to infect spodoptera frugiperda (sf9) insect cells. the pharmacological profile and the status of several post-translational modifications of the glur6 protein were analyzed. saturation analyses of [3h]kainic acid binding demonstrated that glur6 expr ...	1994	8157071
association of peroxisome proliferator-activated receptor and hsp72.	in an effort to understand the relationship between a 72-kda heat shock protein (hsp72) and peroxisome proliferator-activated receptors (ppars), we have characterized their interaction using clofibric acid-sepharose chromatography and co-immunoprecipitation with antisera raised against either rat ppar (rppar) or hsp72. first, we observed that both rppar and hsp72 elute in a clofibrate-dependent manner from the clofibric acid-sepharose matrix. second, we found that immunoprecipitation of either p ...	1994	8132576
amphibian allantoinase. molecular cloning, tissue distribution, and functional expression.	the chain of enzymes necessary to convert uric acid to its metabolic products urea and glyoxylic acid in vertebrates is truncated through the successive loss of allantoicase, allantoinase, and urate oxidase during phylogenetic evolution. previous studies have assigned the localization of both urate oxidase and allantinase to the peroxisome in the amphibian liver. this study reports the cloning of a cdna encoding bullfrog (rana catesbeiana) allantoinase, an enzyme that converts allantoin to allan ...	1994	8163532
expression of ricin b chain in spodoptera frugiperda.	dna encoding ricin b chain was derived from preproricin genomic dna and ligated into the baculovirus transfer vector, pacgp67a. co-transfection of spodoptera frugiperda sf9 cells with baculogold dna was followed by limiting dilution purification of recombinant baculovirus. infection of sf9 cells at a multiplicity of infection of 1 in the presence of 25 mm lactose produced 3 mg/l of soluble, glycosylated, 34 kda protein immunoreactive with monoclonal and polyclonal antibodies to ricin b chain. th ...	1994	7980448
production, purification, and characterization of recombinant maspin proteins.	maspin, a novel mammary serine protease inhibitor, was shown to have tumor suppressing activity (zou, z., anisowicz, a., hendrix, m. j. c., thor, a., neveu, m., sheng, s., rafidi, k., seftor, e., and sager, r. (1994) science 263, 526-529). in this paper, we report the production of recombinant glutathione s-transferase-maspin fusion protein, expressed in the bacterium escherichia coli, and recombinant maspin, expressed in the insect spodoptera frugiperda cells. the fusion protein was purified by ...	1994	7983035
iodinated 2-aminotetralins and 3-amino-1-benzopyrans: ligands for dopamine d2 and d3 receptors.	in developing selective ligands for dopamine d2 and d3 receptors, several iodinated 2-aminotetralins and 3-amino-1-benzopyrans, trans-7-hydroxy-2-[n-(3'-iodo-2'- propenyl)amino]tetralin (1), trans-monohydroxy-2-[n-propyl-n-(3'-iodo-2'-propenyl)amino]tetralin (7-, 5-, and 6-oh-pipat) (2, 3, and 4), and trans-monohydroxy-3,4-dihydro-3-[n-propyl-n-(3'-iodo-2'- propenyl)-amino]-2h-1-benzopyran (6- and 8-oh-benzopyrans) (5 and 6), were prepared. these compounds were evaluated for their binding profil ...	1994	7990123
human 17 beta-hydroxysteroid dehydrogenase: overproduction using a baculovirus expression system and characterization.	estrogenic 17 beta-hydroxysteroid dehydrogenase (17 beta-hsd) plays a pivotal role in the synthesis of estrogens. we overproduced human placental estrogenic 17 beta-hsd using a baculovirus expression system for the study of the enzyme mechanism. a cdna encoding the entire open reading frame of human 17 beta-hsd was inserted into the genome of autographa californica nuclear polyhedrosis virus and expressed in spodoptera frugiperda (sf9) insect cells. metabolic labeling and western blot analysis u ...	1994	7918113
the nonstructural glycoprotein of rotavirus affects intracellular calcium levels.	rotavirus infection of monkey kidney cells has been reported to result in a significant increase in the concentration of intracellular calcium. this increase in intracellular calcium was associated with viral protein synthesis and cytopathic effects in infected cells. we tested the effect of individual rotavirus proteins on intracellular calcium concentrations in insect spodoptera frugiperda (sf9) cells. insect cells were infected with wild-type baculovirus or baculovirus recombinants that conta ...	1994	8254736
cloning and expression of the nucleocapsid gene of virulent kabete o strain of rinderpest virus in baculovirus: use in differential diagnosis between vaccinated and infected animals.	rinderpest (rp) is a viral disease of ruminants with > 95% morbidity and mortality. we have cloned the cdna of the nucleocapsid (n) gene of the virulent kabete o strain of rinderpest virus (rpvk) and compared its nucleotide and deduced amino acid sequences with those of the n genes of the lapinized strain of rinderpest virus (rpvl), measles virus (mv), and canine distemper virus (cdv). the gene has an open reading frame of 1575 nucleotides and encodes a protein of 525 amino acids with a molecula ...	1994	8259649
in vivo and in vitro analyses of recombinant baculoviruses lacking a functional cg30 gene.	the cg30 gene of autographa californica nuclear polyhedrosis virus (acmnpv) encodes two sequence motifs, a zinc finger-like motif and a leucine zipper, found in other polypeptides known to be involved in gene regulation. to gain insight into the function of the cg30 product, cg30, we constructed and characterized recombinant viruses lacking a functional cg30 gene. we found that cg30 mutants had no striking phenotype in cell lines derived from spodoptera frugiperda or trichoplusia ni or in t. ni ...	1994	8289348
high-level expression and purification of secreted forms of herpes simplex virus type 1 glycoprotein gd synthesized by baculovirus-infected insect cells.	two forms of herpes simplex virus glycoprotein gd were recombined into autographa californica nuclear polyhedrosis virus (baculovirus) and expressed in infected spodoptera frugiperda (sf9) cells. each protein was truncated at residue 306 of mature gd. one form, gd-1(306t), contains the coding sequence of patton strain herpes simplex virus type 1 gd; the other, gd-1(qaat), contains three mutations which eliminate all signals for addition of n-linked oligosaccharides. prior to recombination, each ...	1994	8289380
functional mapping of regions of the autographa californica nuclear polyhedrosis viral genome required for dna replication.	previous results showed that plasmids containing one of the eight putative origins (ori's) of autographa californica nuclear polyhedrosis virus (acmnpv) are replicated after transfection into spodoptera frugiperda cells if essential trans-acting factors are supplied by acmnpv infection (kool et al., virology, 192, 94-101, 1993a; kool et al., j. gen. virol., in press, 1993b; leisy and rohrmann, virology, 196, 722-730, 1993). in this report a transient complementation assay is described in which f ...	1994	8291249
synapsin iia bundles actin filaments.	synapsins are neuron-specific phosphoproteins associated with small synaptic vesicles in the presynaptic nerve terminal. synapsin i, which has been demonstrated to bundle f-actin in vitro, has been postulated to regulate neurotransmitter release by cross-linking synaptic vesicles to the actin cytoskeleton. to investigate the possible interaction of synapsin ii with actin filaments, we expressed synapsin ii in spodoptera frugiperda and high five insect cells using a recombinant baculovirus. purif ...	1994	7931311
production and characterization of recombinant rat brain-derived neurotrophic factor and neurotrophin-3 from insect cells.	rat brain-derived neurotrophic factor (bdnf) and neurotrophin-3 (nt-3) were engineered for expression in a baculovirus-infected spodoptera frugiperda insect cell system. the bdnf and nt-3 from the culture supernatants were purified by ion-exchange and reverse-phase chromatography to apparent homogeneity. the purification procedure yielded approximately 2 mg of pure rat bdnf or nt-3 per liter of culture supernatant. a single n-terminus only was found for either secreted molecule and was analogous ...	1994	8294909
enzymatic catalysis of disulfide formation.	the formation of disulfide bridges in membrane and secretory proteins occurs during the protein folding process in the endoplasmic reticulum of eukaryotic cells and the periplasm of prokaryotes. the formation of disulfide bridges is facilitated by the thiol-disulfide oxidoreductases, protein disulfide isomerase (pdi) in eukaryotes and dsba in prokaryotes. structure-function analysis of these soluble proteins demonstrates that their active sites are sequences with similarity to the active site of ...	1994	7909462
human pancreatic procolipase expressed in insect cells: purification and characterization.	a cdna clone encoding human pancreatic procolipase was incorporated into a recombinant baculovirus. spodoptera frugiperda insect cells infected with the recombinant baculovirus secreted procolipase into the medium, which could be isolated in a single step by immunoaffinity chromatography. the highly purified protein reactivated human pancreatic lipase in a concentration-dependent fashion and was efficiently converted to colipase by limited trypsin digestion. this expression system is suitable fo ...	1994	7858428
spodoptera frugiperda sf-9 cells nuclear factor binds to a specific sequence on the baculovirus polyhedrin promoter.	the promoter of the baculovirus polyhedrin gene is used for the heterologous expression of proteins in the baculovirus expression system. the polyhedrin gene of baculovirus is a very late gene that is hyper-expressed 18-24 h post-infection (p.i). we analyzed the dna-binding activity of the polyhedrin promoter using gel mobility shift analysis, dnasei footprint analysis and uv cross-linking experiments. specific dna-binding activity was found in uninfected cells. specific binding activity was als ...	1994	8191787
detection of a putative 30-kda ligand of the cluster-2 antigen.	the cluster-2 antigen, also called egp-2, is a 38-kda transmembrane glycoprotein with a distribution that is largely confined to human epithelial cells and their derived carcinomas. monoclonal antibodies (mabs) directed against egp-2 have been extensively studied as anti-tumor agents, yet the function of the antigen is not known. in the present study we used a biotinylated recombinant soluble derivative of the egp-2 (segpbio) as a probe to detect a possible egp-2 ligand, using various carcinoma ...	1994	8194899
eukaryotic expression of recombinant human centromere autoantigen and its use in a novel elisa for diagnosis of crest syndrome.	the cdna coding for the human full-length centromere protein b (cenp-b) was isolated from a liver cdna library by oligonucleotide screening and extended at the 5' and 3' ends by linker addition. the cdna was inserted into a modified baculovirus transfer vector which mediated high-level expression of recombinant human cenp-b with a histidine-hexapeptide as affinity ligand at its n-terminus in infected spodoptera frugiperda (sf9) insect cells. based on the histidine-hexapeptide moiety, the recombi ...	1994	8198697
cloning and production of functional active recombinant hepatitis b virus surface antigen binding protein.	endonexin ii present on the surface of human hepatocytes has recently been identified as a hepatitis b virus surface antigen (hbsag) binding protein. a full-length cdna clone encoding human endonexin ii was isolated from a human liver cdna library and was placed under the control of the polyhedrin promoter of autographa californica nuclear polyhedrosis virus (acnpv). infection of spodoptera frugiperda cells with recombinant virus resulted in the production of high amounts of recombinant protein. ...	1994	7999073
expression and characterization of human dopamine d2 receptor in baculovirus-infected insect cells.	multiple types of dopamine d2-like receptors (d2, d3, d4) have been identified. differences in pharmacology among these receptors may have profound clinical ramifications for the treatment of psychosis. analysis of the structure and function of their binding sites requires a source of large amounts of receptor, uncontaminated by the other types of d2-like receptor. we engineered a recombinant baculovirus containing the human d2 receptor cdna (drd2) to express this receptor in insect cells. spodo ...	1994	8182575
correct in vivo processing of a chimeric ubiquitin-proapolipoprotein a-i fusion protein in baculovirus-infected insect cells.	the cdna coding for human proapolipoprotein a-i was expressed as a ubiquitin fusion under the control of the polyhedrin promoter in baculovirus-infected sf9 spodoptera frugiperda insect cells. the fusion protein was expressed at high level and was quantitatively cleaved in vivo. the cleaved product was purified and its n-terminal amino acid sequence was established. the data showed that authentic proapolipoprotein a-i has been produced, and thus demonstrated the existence in spodoptera frugiperd ...	1994	7764450
the porcine thromboxane synthase-encoding cdna: sequence, mrna expression and enzyme production in sf9 insect cells.	a full-length cdna encoding porcine thromboxane synthase (ts) was isolated and sequenced. the open reading frame encodes a 534-amino acid (aa) protein (m(r) 60,451) which shares more than 75% identity with ts from other species and is 30% homologous to several enzymes of the cytochrome p-450 iii family. sequence comparison among porcine (p), human (h), and murine (m) ts indicated conservation of eight cys residues and one putative n-glycosylation site. several highly conserved regions were ident ...	1994	8144035
human monocyte chemoattractant protein-1 expressed in a baculovirus system.	human monocyte chemoattractant protein-1 (hmcp-1) was produced using a baculovirus system. the hmcp-1 cdna was inserted into the genomic dna of autographa californica nuclear polyhedrosis virus (acnpv) using a transfer vector, pjvp10z. spodoptera frugiperda insect cells, which were infected with this recombinant virus, secreted recombinant hmcp-1 (re-hmcp-1) at the level of 10-20 micrograms/ml of culture medium. this product was shown to chemoattract monocytes. three distinct bands of 11, 11.5 a ...	1994	8144036
dengue type-2 virus envelope protein made using recombinant baculovirus protects mice against virus challenge.	the gene coding for the envelope (e) glycoprotein of dengue-2 virus was cloned into baculovirus (autographa californica nuclear polyhedrosis virus). the recombinant virus contained the entire e protein gene, preceded by 38 nucleotides from the end of the prematrix glycoprotein gene and followed by the first 83 nucleotides of nonstructural protein 1. when expressed in spodoptera frugiperda (sf9) cells, approximately 1 mg of recombinant e antigen was made per 10(9) cells. this antigen reacted with ...	1994	8147490
optimization of the growth conditions of sf21 insect cells for high-density perfusion culture in stirred-tank bioreactors.	spodoptera frugiperda insect cells (iplb-sf21-ae) (sf21), infected with baculovirus expression vectors during their exponential growth phase, are commonly used to produce a variety of heterologous recombinant proteins. in the present study the culture conditions of these insect cells were studied to establish high-density suspension cultures with prolonged exponential growth phases. the sf21 cells were grown in 125-ml spinner flasks using five different culture media supplemented with 5% fetal c ...	1994	7764890
reconstitution of recombinant 33-kda subunit of the clathrin-coated vesicle h(+)-atpase.	evidence suggests that the atp hydrolytic sector of the clathrin-coated vesicle proton-translocating atpase is composed of four subunits of molecular masses of 70, 58, 40, and 33 kda (xie, x. s., and stone, d. k. (1988) j. biol. chem. 263, 9859-9867). we have now expressed recombinant 33-kda polypeptide in escherichia coli and in spodoptera frugiperda (sf9) cells. this subunit, renatured and purified from both sources, lacks intrinsic atpase activity. co-reconstitution of these recombinant 33-kd ...	1994	8157666
continuous insect cell (sf-9) culture with aeration through sparging.	the continuous growth of spodoptera frugiperda sf-9 cells in a 250-ml blown-glass jacketed spinner flask under a direct air sparging environment was investigated. even at 220 ml working volume (about 90% of total volume), this spinner flask provided good mixing and oxygenation as demonstrated by a higher cell density compared with fermentor cultures. this eliminates a common limitation of the traditional spinner flask, namely much lower cell density at high working volume. furthermore, this spin ...	1994	7764966
evaluation of monitoring approaches and effects of culture conditions on recombinant protein production in baculovirus-infected insect cells.	the baculovirus infection process of spodoptera frugiperda (sf9) insect cells in oxygen-controlled bioreactors in serum-free medium was investigated using a recombinant autographa californica (acnpv) virus expressing beta-galactosidase enzyme as a model system. a variety of monitoring techniques including trypan blue exclusion, fluorescent dye staining, oxygen uptake rate (our) measurements, and glucose consumption were applied to infected cells to determine the best way of evaluating cell integ ...	1994	7765930
functional expression of rat peroxisomal acyl-coa oxidase in spodoptera frugiperda cells.	a cdna coding the 661-residue rat peroxisomal fatty acyl-coa oxidase (acox) has been constructed and expressed as catalytically active protein in spodoptera frugiperda (sf9) insect cells using baculovirus expression system. recombinant viral clones were purified and the expressed protein was identified by immunoblotting and catalytic activity. in the rat liver, acox consists of three polypeptide components a, b and c, with relative molecular mass of 72 kda, 51 kda and 21 kda, respectively. in sf ...	1994	8166685
characterization of [125i](r)-trans-7-hydroxy-2-[n-propyl-n-(3'-iodo-2'-propenyl)amino] tetralin binding to dopamine d3 receptors in rat olfactory tubercle.	[125i](r,s)-trans-7-hydroxy-2-[n-propyl-n-(3'-iodo-2'-propenyl)- amino]tetralin ([125](r,s)-trans-7-oh-pipat) has been shown to bind with high affinity to dopamine d3 receptors expressed in spodoptera frugiperda cells. no specific binding was seen in spodoptera frugiperda cells expressing a high density of d2 receptors. it was therefore, suggested that [125i] (r,s)-trans-7-oh-pipat selectively labels d3 receptors. in the present study, saturation binding of [125i](r)-trans-7-oh-pipat to membrane ...	1994	8114008
location of agonist-dependent-phosphorylation sites in the third intracellular loop of muscarinic acetylcholine receptors (m2 subtype).	muscarinic acetylcholine receptors (machr, human m2 subtype) expressed in sf9 (spodoptera frugiperda) cells using the baculovirus system were purified and subjected to phosphorylation by a machr kinase, which was partially purified from porcine cerebrum. two bands with apparent molecular masses of 59 kda and 39 kda as determined by sds/page were found to be phosphorylated in an agonist-dependent manner. both bands were labeled by the irreversible muscarinic ligand [3h]propylbenzilylcholine musta ...	1994	8119296
complementation of human immunodeficiency virus (hiv-1) gag particle formation.	the human immunodeficiency virus gag precursor protein pr55gag exhibits the ability of particle assembly when expressed using recombinant baculoviruses. in order to delineate the sequences required for particle formation, two mutants of gag (d1 and d2) were constructed in which 10 amino acids within the ca domain were deleted. both mutants yielded stable high levels of gag antigen following expression in spodoptera frugiperda insect cells. electron microscopy of sections through infected cells r ...	1994	8122369
development of a recombinant enzyme-linked immunosorbent assay for detection of antibodies against epstein-barr virus nuclear antigens 2a and 2b.	the baculovirus expression system was used to produce full-length epstein-barr virus nuclear antigens (ebnas) 2a and 2b. recombinant baculoviruses that contained the ebna-2a- and ebna-2b-encoding sequences were constructed. the proteins were expressed in spodoptera frugiperda sf-9 cells infected with the recombinant viruses and were characterized by using monoclonal and human polyclonal antibodies by immunoblotting and immunofluorescence techniques. partially purified extracts of the ebna-2a- an ...	1994	8126164
metallopanstimulin gene product produced in a baculovirus expression system is a nuclear phosphoprotein that binds to dna.	the protein product of the human metallopanstimulin (mps-1) gene was produced in the insect cell line spodoptera frugiperda (sf9) using the baculovirus expression vector system autographa californica nuclear polyhedrosis virus (acnpv). when a cloned mps-1 complementary dna sequence was inserted into the acnpv viral genome downstream from the promoter of the polyhedrin gene, a polypeptide with an apparent molecular weight of approximately 10,000 was observed in extracts of infected sf9 cells. thi ...	1994	7986747
molecular characterization of a strain-specific repeated dna sequence in the fall armyworm spodoptera frugiperda (lepidoptera: noctuidae).	the fall armyworm moth, spodoptera frugiperda, is a migratory species composed of sympatric corn and rice strains. the strains are indistinguishable in morphology but can be recognized by molecular markers. we have cloned and characterized seven monomer units of a repeated dna sequence, called fr, which is found exclusively in the genome of the rice strain individuals. the 189 bp fr units are tandemly organized in arrays longer than 30 kb. female individuals possess over 100-fold more of the fr ...	1994	7987522
baculovirus expression of non-structural protein ns2 and core protein vp7 of african horsesickness virus serotype 3 and their use as antigens in an indirect elisa.	non-structural protein ns2 and core protein vp7 of african horsesickness virus serotype 3 (ahsv3) were expressed in spodoptera frugiperda cells by recombinant baculoviruses containing the relevant genes. these proteins were purified and analysed by polyacrylamide gel electrophoresis and western blot. ns2 and vp7 were used separately as antigens in an indirect elisa for the detection of ahsv antibodies. both antigens cross-reacted with hyperimmune guinea-pig antisera to infected cell lysates of a ...	1994	7989441
preparation and characterization of human rheumatoid arthritic synovial fluid phospholipase a2 produced by recombinant baculovirus-infected insect cells.	we prepared human rheumatoid arthritic synovial fluid phospholipase a2 (pla2) [ec 3.1.1.4] from insect cells infected with a recombinant baculovirus. the pla2 dna was designed, changing the original codons to those used frequently in the polyhedrin gene. sixteen oligo-deoxynucleotides ranging from 40 to 70 nucleotides were chemically synthesized and then assembled to form the whole pla2 gene. the gene thus synthesized was then placed under the control of the polyhedrin promoter of autographa cal ...	1994	7798190
protective efficacy in mice of a secreted form of recombinant dengue-2 virus envelope protein produced in baculovirus infected insect cells.	we constructed a recombinant baculovirus encoding a dengue (den)-2 virus envelope glycoprotein truncated of 102 amino acids (aa) at its c-terminus (d2e delta 102). the production, processing and transportation of the recombinant protein in baculovirus-infected spodoptera frugiperda (sf9) cells and its immunogenic properties in mice were compared to those of a previously characterized recombinant den-2 e-protein with a 71aa c-terminal truncation (d2e delta 71). both proteins were transported thro ...	1994	7826210
altered glycosylation and selected mutation in recombinant human complement component c9: effects on haemolytic activity.	recombinant wild-type and mutated forms of human complement component c9 have been synthesized in baculovirus-infected insect cells. wild-type recombinant c9 was indistinguishable from native c9, as judged by haemolytic activity, trypsin and alpha-thrombin digestion, reaction with antibodies to c9, enzymatic deglycosylation to the same core size and polymerization in the presence of zn2+. replacement of the native signal peptide with the honey-bee melittin signal peptide, and replacement of spod ...	1994	7835977
plaque assay and replication of tipula iridescent virus in spodoptera frugiperda ovarian cells.	a plaque assay was developed for the study of tipula iridescent virus (tiv) replication using a cell line derived from the fall army worm spodoptera frugiperda (sf9). infection and plaque formation were monitored with time by phase contrast microscopy, video and fluorescent light microscopy. structure of virions, viroplasmic centres and organelles of infected cells were examined by transmission electron microscopy (tem). after 4 h postinfection, plaques were visibly detected within the cell mono ...	1994	7839010
effects of co-expressing chaperone bip on functional antibody production in the baculovirus system.	the assembly pathway of the insect cell spodoptera frugiperda (sf-9) was engineered to include expression of the murine chaperone immunoglobulin heavy chain binding protein (bip) using the baculovirus vector. the impact of bip coexpression on the production and secretion of functional and soluble recombinant immunoglobulin igg levels was evaluated. recombinant bip was found to associate specifically with immunoglobulins in immunoprecipitation studies. coinfection of insect cells with a bip-conta ...	1994	7858430
the structure and function of the first component of complement: genetic engineering approach (a review).	the availability of cdna and genomic clones for the subcomponents of c1, as well as the recognition of the modular organization of serine-proteases have opened up exciting new possibilities for approaching structural problems. in this review the latest achievements of combined protein engineering, functional and structural studies are summarized. the concept of this research is to construct deletion, point and hybrid mutants of the highly homologous c1r and c1s subcomponents, to reveal the funct ...	1994	7866721
production of measles nucleoprotein in different expression systems and its use as a diagnostic reagent.	measles nucleoprotein has been successfully expressed in three different hosts, bacterial (escherichia coli bl21 de3), insect (spodoptera frugiperda; sf9) and mammalian (primary human fibroblasts) cells, each producing an antigenic protein of m(r) 60 kda. the nucleoprotein produced in all three hosts was used in an elisa for the detection of antibodies to measles virus in a cohort of haemagglutinin-positive or -negative human sera. data produced from baculovirus and adenovirus-based antigens ind ...	1994	7868644
clonal variation in the spodoptera frugiperda iplb-sf21-ae insect cell population.	clones have been isolated from the heterogeneous spodoptera frugiperda iplb-sf21-ae insect cell population. five of these clones, in addition to the parent cell line and the sf9 cell line (another clonal isolate of the parent cell line), have been compared in regards to morphology, growth, budded virus synthesis, and recombinant protein synthesis. no significant differences in cell morphology were found among these cell lines. there was, however, a significant difference in the average cell size ...	1994	7764939
characterization by electrospray mass spectrometry of human ca(2+)-sensitive cytosolic phospholipase a2 produced in baculovirus-infected insect cells.	the 85-kd cytosolic phospholipase a2 (cpla2) is a novel receptor-regulated phospholipase that is thought to initiate the production of inflammatory lipid mediators. since cpla2 is present only in minute amounts (less than 0.01% of total cellular protein) in various cells and tissues, we have used the baculovirus expression system to produce sufficient quantities of cpla2 for structural and functional analysis. the cdna for cpla2 was cloned into a baculovirus expression vector and, upon infection ...	1994	7764328
heterologous expression of the plant proteins strictosidine synthase and berberine bridge enzyme in insect cell culture.	the heterologous expression of cdnas encoding the alkaloid biosynthetic enzymes, strictosidine synthase [ec 4.3.3.2] from rauvolfia serpentina and the berberine bridge enzyme [(s)-reticuline: oxygen oxidoreductase (methylene-bridge-forming), ec 1.5.3.9] from eschscholtzia californica, has been achieved in a cell culture (sf9) of the fall army worm, spodoptera frugiperda, using a baculovirus-based expression system. the expression resulted in the overproduction of each plant enzyme in a catalytic ...	1994	7764480
expression, glycosylation, and phosphorylation of human keratins 8 and 18 in insect cells.	the filament forming ability and post-translational modifications of the human intermediate filaments, keratin polypeptides 8 and 18 (k8/18), were studied in recombinant baculovirus-infected insect (spodoptera frugiperda, sf9) cells. no change in cell morphology was noted after high levels of k8/18 were expressed in sf9 cells coinfected with recombinant virus-containing human k8 and k18. immunofluorescence staining showed that k8/18 expressed in sf9 cells formed somewhat what disorganized and ro ...	1994	7510249
expression and characterization of the alpha 2 subunit of the alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionate (ampa)-selective glutamate receptor channel in a baculovirus system.	using a baculovirus expression vector system, the alpha 2 subunit of the mouse alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionate (ampa)-selective glutamate receptor (glur) channel was expressed in spodoptera frugiperda insect cells. immunoblotting using the antibody made to the synthetic peptide corresponding to the c-terminus of glur alpha 2 and [35s]methionine/[35s]cysteine metabolic radiolabeling revealed the major 102-kda and the minor 98-kda protein bands. metabolic radiolabeling with t ...	1994	7889366
desensitization, phosphorylation and palmitoylation of the human dopamine d1 receptor.	the regulation and post-translational modifications of the human dopamine d1 receptor were studied in the baculovirus-eukaryotic cell expression system. baculovirus constructs containing either the dna encoding the dopamine d1 receptor or a dna encoding a c-myc epitope tagged dopamine d1 receptor (c-myc-dopamine d1 receptor) were used to infect spodoptera frugiperda (sf9) insect cells. expressed dopamine d1 and c-myc-dopamine d1 receptors bound agonists and antagonists with affinities and a rank ...	1994	7515822
baculovirus-mediated expression of the epidermal growth factor-like modules of human factor ix fused to the factor xiiia transamidation site in fibronectin. evidence for a direct interaction between the nh2-terminal epidermal growth factor-like module of factor ixa beta and factor x.	factor ix is a vitamin k-dependent procoagulant zymogen of a serine protease. in the presence of ca2+ the active form of factor ix (factor ixa beta) forms a complex with factor viiia on suitable phospholipid surfaces such as aggregated platelets. this macromolecular complex rapidly activates factor x. we have previously provided data that suggest an interaction between the nh2-terminal epidermal growth factor (egf)-like module of factor ixa beta and the substrate factor x. in an alternative appr ...	1994	7906269
similar ligand binding in recombinant human alpha 2 c2-adrenoceptors produced in mammalian, insect and yeast cells.	ligand binding properties were investigated in recombinant human alpha 2c2-adrenoceptors expressed in three different host systems: shionogi s115 mouse mammary tumour cells, spodoptera frugiperda sf9 insect cells and saccharomyces cerevisiae yeast cells. the expected 43 kda alpha 2c2 protein was visualized with immunoblotting using a polyclonal alpha 2c2-receptor antibody. [3h]rauwolscine binding in cell homogenates or membranes (bmax 3-11 pmol/mg protein; kd approximately 5.5 nm) was inhibited ...	1994	7911432
the alpha-5 segment of bacillus thuringiensis delta-endotoxin: in vitro activity, ion channel formation and molecular modelling.	a peptide with a sequence corresponding to the highly conserved alpha-5 segment of the cry delta-endotoxin family (amino acids 193-215 of bacillus thuringiensis cryiiia [gazit and shai (1993) biochemistry 32, 3429-3436]), was investigated with respect to its interaction with insect membranes, cytotoxicity in vitro towards spodoptera frugiperda (sf-9) cells, and its propensity to form ion channels in planar lipid membranes (plms). selectively labelled analogues of alpha-5 at either the n-terminal ...	1994	7529493
expression of functional metabotropic and ionotropic glutamate receptors in baculovirus-infected insect cells.	an ionotropic glutamate receptor of the kainate subtype (glur6) and a g-protein coupled metabotropic glutamate receptor (mglur1 alpha) were expressed and studied in two insect cell lines: sf9 cells from spodoptera frugiperda and mg1 cells from trichoplusia ni. application of kainate to glur6-infected mg1 cells produced kainate-activated currents. glutamate activation of mglur1 alpha in mg1- and sf9-infected cells caused rapid, transient increases in intracellular calcium levels. this effect was ...	1994	7936400
cystine-rich fish antifreeze is produced as an active proprotein precursor in fall armyworm cells.	recombinant cystine-rich fish antifreeze protein (afp) was produced by fall armyworm cells from a baculovirus expression vector containing sea raven afp cdna. the natural signal sequence encoded in the cdna directed secretion of the antifreeze into the medium, from where it was recovered and purified to homogeneity. the m(r) of the exported protein (16k), as determined by sds-page, was larger than that (14k) of mature afp isolated from sea raven serum. sequencing of the recombinant afp showed th ...	1994	7945337