Publications

purification and characterization of human salivary-gland prokallikrein from recombinant baculovirus-infected insect cells.	a full-length cdna encoding human salivary-gland preprokallikrein was inserted into the baculovirus autographa californica nuclear polyhedrosis virus downstream of the polyhedrin promoter. the gene was expressed in transfected spodoptera frugiperda cells and the recombinant product secreted into the culture medium. by alternating anion-exchange chromatography and gel-filtration steps, twice repeated, prokallikrein was purified to homogeneity, which was confirmed by amino acid analysis and n-term ...	1992	1587272
rat urate oxidase produced by recombinant baculovirus expression: formation of peroxisome crystalloid core-like structures.	urate oxidase (ec 1.7.3.3), which catalyzes the oxidation of uric acid to allantoin, is present in most mammals but absent in humans and hominoid primates. in rats and most other mammals that catabolize uric acid to allantoin, this enzyme is localized within the crystalloid cores of peroxisomes present in liver parenchymal cells. to determine whether urate oxidase forms these crystalloid cores or whether core-forming protein(s) exist in association with urate oxidase, a baculovirus expression ve ...	1992	1594592
secretion of a functional soluble form of neutral endopeptidase-24.11 from a baculovirus-infected insect cell line.	neutral endopeptidase (nep; ec 3.4.24.11) is an integral membrane protein found at the plasma membrane of many cell types. a secreted form of nep (sec-nep) was recently obtained by transfection of cos-1 cells with a recombinant expression vector consisting of the cdna encoding the signal peptide of pro-opiomelanocortin fused in-frame to the cdna sequence of the complete ectodomain of rabbit nep [lemay, waksman, roques, crine & boileau (1989) j. biol. chem. 264, 15620-15623]. in order to produce ...	1992	1599410
overproduction and large-scale purification of the human poly(adp-ribose) polymerase using a baculovirus expression system.	we have overproduced the full-length human poly(adp-ribose) polymerase (parp) in spodoptera frugiperda (sf9) cells using a baculovirus expression vector system. approx. 20 mg of purified protein from 5 x 10(8) sf9 cells were obtained by a simple three-step purification procedure including 3-aminobenzamide affinity chromatography. the recombinant protein (reparp), which migrates as a unique 116-kda band on sds-polyacrylamide gels, was identified as parp by western blotting using either polyclonal ...	1992	1601310
dissimilar expression of autographa californica multiple nucleocapsid nuclear polyhedrosis virus polyhedrin and p10 genes.	the temporal expression of the autographa californica multiple nucleocapsid nuclear polyhedrosis virus polyhedrin and p10 genes in spodoptera frugiperda cells was studied using virus recombinants in which either gene was replaced by the juvenile hormone esterase (jhe) gene of heliothis virescens. the jhe served as a highly specific and sensitive reporter for gene expression. activation of the p10 gene followed a pattern different to that of polyhedrin. the p10 gene was activated a few hours earl ...	1992	1607866
hepatitis b virus polymerase gene: expression of the long open reading frame using the baculovirus expression system.	a recombinant baculovirus was constructed containing a copy of the hepatitis b virus (hbv) genome which was inserted to produce an in-frame fusion of the precore (pre-c) coding region with the first 11 amino acids of the polyhedrin gene. the recombinant baculovirus expressed the 25k pre-c protein and two novel proteins, of approximately 93k and 72k. both the 93k and 72k proteins are recognized by an anti-polymerase monoclonal antibody. northern blot analysis of the mrna produced during infection ...	1992	1607871
acidic ph induces fusion of cells infected with baculovirus to form syncytia.	the enveloped baculovirus insect cell system has been used extensively for expression of recombinant proteins, including viral fusion proteins. we tested wild-type baculovirus for endogenous fusion protein activity. syncytia formation, dye transfer, and capacitance changes were observed after incubating infected spodoptera frugiperda cells in acidic media, consistent with fusion protein activity. only a short acidic pulse of 10 s is needed to trigger syncytia formation. identical results were ob ...	1992	1618326
foreign gene expression by a baculovirus vector with an expanded host range.	a nuclear polyhedrosis virus (npv) (baculoviridae)-based gene expression system was improved by dna recombination. the bmn cell line established from bombyx mori and the sf21 cell line established from spodoptera frugiperda are non-permissive for autographa californica multicapsid npv (acmnpv) and b. mori npv (bmnpv) replication, respectively. after cotransfection of acmnpv dna and bamhi-digested bmnpv dna into sf21 cells, progeny viruses were isolated by plaque purification on bmn cell monolaye ...	1992	1629707
expression of recombinant soluble and membrane-bound catechol o-methyltransferase in eukaryotic cells and identification of the respective enzymes in rat brain.	the rat and human recombinant soluble and membrane-bound catechol o-methyltransferase (s- and mb-comt, respectively) were expressed using mammalian and baculovirus vectors. low levels of rat and human s-comt polypeptides were detected by immunoprecipitation in k-562 cell lines transfected with the s-comt vectors. from k-562 cells transfected with the rat mb-comt construct, both s- and mb-comt recombinant proteins were detected by a rat comt-specific anti-serum. infection of lepidopteran spodopte ...	1992	1633830
baculovirus expression and characterization of catalytically active horseradish peroxidase.	studies of horseradish peroxidase (hrp), a prototypical enzyme, have provided much of the information that is available on the mechanisms and functions of hemoprotein peroxidases. hrp itself is widely used in biotechnological applications. further progress in defining the structure and function of the enzyme, however, requires its expression in a heterologous system. we report here baculovirus-mediated, high yield expression of a synthetic gene for hrp in spodoptera frugiperda cell culture. expr ...	1992	1637184
baculovirus-expressed rabies virus m1 protein is not phosphorylated: it forms multiple complexes with expressed rabies n protein.	the rabies n, m1, m2, and g antigens have been expressed in spodoptera frugiperda cells from single gene expression vectors or dual gene vectors (n/m1 or m2/g) using the baculovirus system. although n protein was phosphorylated, no evidence for m1 phosphorylation was obtained. n-m1 complexes were formed in vivo using dual infections or the coexpression vectors, as well as in vitro in mixing experiments. the free or complexed rabies n and m1 proteins reacted with available monoclonal and polyclon ...	1992	1641989
expression and purification of functional g protein alpha subunits using a baculovirus expression system.	the alpha subunits of the heterotrimeric guanine nucleotide-binding proteins gi1, gi2, gi3, g0, and gs have been overexpressed in sf9 cells using a baculovirus expression system. the gi1 alpha, gi2 alpha, gi3 alpha, and g0 alpha have been purified to homogeneity from infected spodoptera frugiperda (sf9) cells and characterized. yields of up to 1.8 mg of purified recombinant g alpha have been obtained from 300-ml cultures of infected cells. the recombinant alpha subunits are myristoylated and are ...	1992	1730649
synthesis and toxicity of full-length and truncated bacterial cryivd mosquitocidal proteins expressed in lepidopteran cells using a baculovirus vector.	full-length (72k) and truncated (61k) cryivd mosquitocidal proteins of bacillus thuringiensis (bt) were expressed in spodoptera frugiperda cells and larvae of trichoplusia ni using a baculovirus vector to investigate the role of cryivd peptides in toxicity as well as to evaluate further the baculovirus/lepidopteran system for expressing bt proteins. the cryivd genes were inserted into the autographa californica multinucleocapsid nuclear polyhedrosis virus (acmnpv) under control of the polyhedrin ...	1992	1730944
expression of the mouse c-abl type iv proto-oncogene product in the insect cell baculovirus system.	the cellular gene c-abl is the normal homologue of the transforming gene (v-abl) within the genome of the abelson leukaemia virus. the cdna sequence coding for the cellular form of the murine abl gene (c-abl type iv) has been inserted into the baculovirus transfer vector, pac36c, so that the c-abl gene is under the control of the polyhedrin promoter of autographa californica nuclear polyhedrosis virus (acnpv). spodoptera frugiperda cells infected with the recombinant transfer vector in the prese ...	1992	1737071
baculovirus recombinant expressing a secreted form of a transmembrane carcinoma-associated antigen.	ga733-2 is a monoclonal antibody-defined, 40-kda glycoprotein antigen that is associated with carcinomas of various origins. hydrophobicity analysis of the protein sequence predicted by complementary dna (cdna) has suggested that the ga733-2 antigen is a type i membrane protein. in this study, the polymerase chain reaction was used in a strategy to omit cdna sequences for the transmembrane and cytoplasmic domains, thereby converting the extracellular domain into a secretory protein. full-length ...	1992	1737342
expression of the saccharomyces cerevisiae kex2p endoprotease in inset cells. evidence for a carboxy-terminal autoprocessing event.	the pheromone-processing kex2p endoprotease of saccharomyces cerevisiae has been difficult to characterize due to its low level of expression in yeast cells. to overcome this problem, we have overexpressed kex2p using the baculovirus/insect cell expression system. spodoptera frugiperda sf9 insect cells infected with a recombinant baculovirus, containing the complete kex2 gene which encodes the kex2p protease (814 amino acids), accumulate an 120-kda functional form of the enzyme. the inhibition p ...	1992	1740121
baculovirus gene me53, which contains a putative zinc finger motif, is one of the major early-transcribed genes.	we have been investigating the first viral genes to be transcribed in spodoptera frugiperda cells after the infection with autographa california nuclear polyhedrosis virus in order to identify regulatory proteins required for the activation of early and/or late gene transcription. by screening a cdna library from polyadenylated rna transcribed at 1 h postinfection, we identified a gene which is designated me53. this newly identified gene is located upstream of the late p74 gene and forms a diver ...	1993	8093490
human papillomavirus type 16 e6, e7 and l1 and type 18 e7 proteins produced by recombinant baculoviruses.	proteins derived from the e6, e7 and l1 orfs of hpv16 and the e7 orf of hpv18 were produced in insect cells using a baculovirus expression system. hpv orfs were inserted into baculovirus transfer vectors pacym1 or pvl1393/2, and recombinant baculoviruses isolated using a combination of limiting dilution and plaque assay. using hpv-specific antisera and monoclonal antibodies hpv proteins were identified in lysates of spodoptera frugiperda (sf-21) cells infected with hpv-recombinant baculovirus. i ...	1993	8106603
processing of asparagine-linked oligosaccharides in insect cells. n-acetylglucosaminyltransferase i and ii activities in cultured lepidopteran cells.	the levels of beta 1,2-n-acetylglucosaminyltransferase (glcnac-t) i and ii activities in cultured cells from bombyx mori (bm-n), mamestra brassicae (izd-mb-0503) and spodoptera frugiperda (sf-9 and sf-21) were investigated. apart from initial experiments with man alpha-3(man alpha 1-6)-man beta 1-o(ch2)8cooh3 and 3h-labelled udp-glcnac as substrates, glcnac-t i activity was measured with a non-radioactive hplc method using pyridylaminated man3-glcnac2 and man5glcnac2 as acceptor oligosaccharides ...	1993	8130393
cloning and expression of a rat neuronal nitric oxide synthase coding sequence in a baculovirus/insect cell system.	a dna sequence encoding rat neuronal no synthase (nnos) was isolated and cloned into the baculovirus expression vector pvl1393 to generate pvlrbnos. transfection of spodoptera frugiperda sf-21 cells with the construct pvlrbnos resulted in the synthesis of high levels of neuronal no synthase. analysis of the expression pattern revealed soluble, enzymatically active no synthase in the cytoplasm of cell extracts. active enzyme could also be purified from culture supernatants using 2'-5' adp sepharo ...	1993	7504478
lepidopteran-specific crystal toxins from bacillus thuringiensis form cation- and anion-selective channels in planar lipid bilayers.	previous studies in our laboratory have shown that cryic, a lepidopteran-specific toxin from bacillus thuringiensis, triggers calcium and chloride channel activity in sf-9 cells (spodoptera frugiperda, fall armyworm). chloride currents were also observed in sf-9 membrane patches upon addition of cryic toxin to the cytoplasmic side of the membrane. in the present study the ability of activated cryic toxin to form channels was investigated in a receptor-free, artificial phospholipid membrane syste ...	1993	7681482
high level expression and phosphorylation of hepatitis b virus polymerase in insect cells with recombinant baculoviruses.	the hepatitis b virus polymerase open reading frame, as well as various subdomains of polymerase, was expressed in insect cells using the recombinant baculovirus expression system. full-length polymerase was expressed at very low levels in a spodoptera frugiperda cell line, the amino-terminal domain of polymerase was expressed at high levels, and other constructs were expressed at intermediate levels. infections of a trichoplusia ni cell line with the same recombinant baculoviruses resulted in h ...	1993	7683161
expression of a recombinant baculovirus for vitronectin in insect cells: purification, characterization of post-translational modifications and functional studies of the recombinant protein.	vitronectin (vn) is a cell attachment glycoprotein present in plasma and the extracellular matrix that also has multiple regulatory roles in the complement, coagulation, and fibrinolytic systems. vn stabilizes plasminogen activator inhibitor-1 (pai-1) with potential binding sites reported at both the somatomedin b and heparin binding domains. in order to perform detailed structure-function analyses of vn, we have produced and expressed a recombinant baculovirus containing the cdna of human vn. r ...	1993	7688496
expression of the glycosylphosphatidylinositol-linked complement-inhibiting protein cd59 antigen in insect cells using a baculovirus vector.	cd59 antigen (cd59) is a glycosylphosphatidylinositol (gpi)-linked membrane glycoprotein which protects human cells from complement-mediated lysis. here we report the expression of functionally active cd59 in spodoptera frugiperda insect cells using a baculovirus vector. recombinant cd59 was expressed abundantly on the surface of the insect cells and protected the cells from lysis by human complement. the protein was released from the cell surface by treatment with phosphatidylinositol-specific ...	1993	7694573
ion channel activity of n-terminal fragments from cryia(c) delta-endotoxin.	proteolytically-derived amino-terminal fragments from the bacillus thuringiensis delta-endotoxin, cryia(c), demonstrated ion channel activity in two in vitro assays in the absence of any membrane receptors. a 24.5 kda fragment (from spodoptera frugiperda digestive proteolysis) formed cation-selective channels (10mv for a 3:1 salt gradient) in planar lipid bilayers up to several hundred picosiemen conductance. a 21.5 kda fragment doublet (from alkaline hydrolysis) also showed large conductance in ...	1993	7694582
effect of temperature and oxygen on cell growth and recombinant protein production in insect cell cultures.	the effect of temperature and o2 saturation on the production of recombinant proteins beta-galactosidase and human glucocerebrosidase by spodoptera frugiperda cells (sf9) infected with recombinant autographa californica nuclear polyhedrosis virus was investigated. the rates of cell growth, glucose consumption, o2 consumption and product expression were measured at temperatures between 22 degrees c and 35 degrees c. the results indicated that possible o2 limitation may be alleviated without compr ...	1993	7763472
insect cell hosts for baculovirus expression vectors contain endogenous exoglycosidase activity.	four different insect cell lines that can be used as hosts for baculovirus infection were assayed for the presence of endogenous exoglycosidases. all four cell lines, derived from spodoptera frugiperda, trichoplusia ni, bombyx mori, or malacosoma disstria, contained n-acetyl-beta-glucosaminidase, n-acetyl-beta-galactosaminidase, beta-galactosidase, and sialidase activities. exoglycosidase activities were found in cell lysates as well as cell-free supernatants from uninfected and wild-type baculo ...	1993	7763590
biotechnological aspects of the production of human pro-kallikrein using the acnpv-baculovirus-expression system.	we have investigated large scale production processes (up to 2 liters) of recombinant proteins using the baculovirus expression system in order to optimize the product yields. experiments using cell lines of spodoptera frugiperda (sf9) and mamestra brassicae (izd-mb0503) were performed to show the different production capacities of the cell lines. the influence of the infection at different cell densities is described. beyond that, tc100-, ipl41- and serum-free ipl41-medium were compared to demo ...	1993	7763638
host-specific transcription of baculovirus genes.	we examined the differential replication of autographa californica multicapsid nuclear polyhedrosis virus (acmnpv; family: baculoviridae) in spodoptera frugiperda (sf) and bombyx mori (bm) cell lines. our previous studies have shown that infection of bm cells is abortive; infectious progeny is not produced, and although virogenic stroma is produced, complete virions or inclusion bodies are not assembled. we designed the present study to determine the extent of infected-cell-specific polypeptides ...	1993	7763702
optimization of growth methods and recombinant protein production in bti-tn-5b1-4 insect cells using the baculovirus expression system.	a novel insect cell line from trichoplusia ni, bti-tn 5b1-4 (tn 5), was compared to spodoptera frugiperda, sf 9, cells for production of two recombinant secreted proteins: truncated epstein-barr viral attachment protein (ebv gp105) and truncated, soluble tissue factor (stf). under optimum conditions for both cell lines, tn 5 cells produced 28-fold more secreted stf than sf 9 cells, respectively, on a per cell basis. the total production of gp105 was similar for the two cell lines. however, tn5 c ...	1993	7764044
fed-batch culture of insect cells: a method to increase the yield of recombinant human nerve growth factor (rhngf) in the baculovirus expression system.	a fed-batch method was developed which increased the density of insect cells (spodoptera frugiperda, sf-9 cells) in suspension culture and the feeding of nutrients improved the yield of a recombinant protein produced by a baculovirus expression system. analysis of spent medium samples indicated that depletions of glucose and glutamine correlated with the retardation of cell growth. feeding of a mixture of nutrients consisting of glucose, glutamine, yeastolate and lipids solution restored the gro ...	1993	7764302
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
identification and immunocharacterization of a 25k structural protein in autographa californica nuclear polyhedrosis virus (acmnpv).	a 25k protein was a major component of the triton x-100 solubilized fraction prepared for the isolation of nuclear matrices from spodoptera frugiperda cells infected with autographa californica nuclear polyhedrosis virus (acmnpv). polyclonal antisera to p25 were produced in rabbits and used in the characterization of p25. western blots of proteins probed with the p25 antiserum demonstrated the presence of the p25 in both the polyhedral-derived and extracellular viral phenotypes. using an indirec ...	1993	8317143
non-homologous recombination between adenovirus and acnpv dna fragments in cell-free extracts from insect spodoptera frugiperda nuclei.	in previous work, we have developed a cell-free system from nuclear extracts of hamster cells to study the mechanism of integrative recombination between adenovirus type 12 (ad12) dna and hamster cell dna (jessberger et al., 1989; tatzelt et al., 1992). we have also demonstrated that in insect cells the left terminal fragment of ad2 dna can insert by non-homologous recombination into the 32.6 to 34.5 map unit (ecori-o) fragment and into other segments of autographa californica nuclear polyhedros ...	1993	8317144
purification and characterization of recombinant histidine-tagged human platelet 12-lipoxygenase expressed in a baculovirus/insect cell system.	a baculoviral expression vector consisting of a sequence encoding a six-histidine tag apposed to the human platelet 12-lipoxygenase cdna, under control of the polyhedrin promoter, was constructed. recombinant 12-lipoxygenase baculoviruses were used to infect spodoptera frugiperda insect cells (sf9). at 54 h post-infection, maximal 12-lipoxygenase activity and protein levels were achieved; the enzyme was purified to apparent homogeneity in a single step by nickel-ion-chelation chromatography in w ...	1993	8319693
isolation, characterization, and proteolysis of human prosaposin, the precursor of saposins (sphingolipid activator proteins).	prosaposin contains separate domains in tandem for four saposins, a, b, c, and d. these mature saposins are produced by limited proteolysis of prosaposin. they are involved in lysosomal hydrolysis of gm1 ganglioside, gluco- and galactocerebrosides, sulfatides, and sphingomyelin and other sphingolipids. prosaposin also exists as a secretory protein in body fluids. in this investigation prosaposin was expressed in spodoptera frugiperda cells (sf9) by infection with baculovirus containing a full le ...	1993	8323276
expression of anticardiolipin cofactor, human beta 2-glycoprotein i, by a recombinant baculovirus/insect cell system.	a full-length cdna coding a human beta 2-glycoprotein i (beta 2-gpi) was introduced into the baculovirus genome to construct a recombinant baculovirus. spodoptera frugiperda (sf9) cells were infected with the recombinant baculovirus. a protein (mol. wt 43,000) reactive with anti-beta 2-gpi antisera was produced in the insect cells and secreted into the culture medium. the recombinant beta 2-gpi was purified from the culture supernatant by sequential cardiolipin (cl)-affinity column chromatograph ...	1993	8324900
phosphorylation of elongation factor-2 from the lepidopteran insect, spodoptera frugiperda.	in mammalian cells, protein synthesis can be regulated at the level of elongation by the phosphorylation of elongation factor 2 (eef-2) by a highly specific ca2+/calmodulin-dependent kinase. in this report, we show that eef-2 from a cell line derived from the insect, spodoptera frugiperda, is a substrate for mammalian eef-2 kinase and that phosphorylation is ca(2+)-dependent. furthermore, two-dimensional peptide mapping shows that the kinase phosphorylates the same sites in spodoptera eef-2 as t ...	1993	8335099
one-step purification and characterization of human pancreatic lipase expressed in insect cells.	a cdna clone encoding the sequence of human pancreatic lipase (hpl) was subcloned into the baculovirus transfer vector pvl1392 and used in co-transfection of spodoptera frugiperda (sf9) insect cells with wild-type autographa californica nuclear polyhedrosis virus (acnpv) dna. a single recombinant protein (50 kda) secreted by sf9 cells was detectable in the culture medium 24 h post-infection using both anti-hpl polyclonal antibodies and potentiometric measurements of lipolytic activity. the expre ...	1993	8335100
a 25-kda protein is associated with the envelopes of occluded baculovirus virions.	antiserum produced against preoccluded virions of the orgyia pseudotsugata multinucleocapsid nuclear polyhedrosis virus (opmnpv) was used to screen an opmnpv lambda gt11 expression library. one of the immunoreactive clones contained an insert that hybridized to a portion of the opmnpv hindiii-p fragment. a 2-kb region of hindiii-p was sequenced and found to contain three open reading frames, each preceded by a late promoter element. the insert from the lambda gt11 clone was derived from the thir ...	1993	8337828
immediate-early baculovirus genes transactivate the p143 gene promoter of autographa californica nuclear polyhedrosis virus.	trans-acting regulatory components of autographa californica nuclear polyhedrosis virus (acmnpv) were studied in a transient assay system for their ability to activate the p143 gene promoter. a region including 502 nt upstream of the aug of the p143 gene was linked to the bacterial chloramphenicol acetyltransferase (cat) gene. the p143 promoter-cat construct was used to identify acmnpv immediate-early gene products required for expression from the p143 gene early promoter. transient expression a ...	1993	8337840
overexpression, characterization, and purification of a recombinant mouse immunophilin fkbp-52 and identification of an associated phosphoprotein.	to gain insight into the structure and function of the immunophilin fkbp-52, a mouse fkbp-52 was overexpressed in spodoptera frugiperda insect cells (sf9 cells) with the baculovirus expression system. the purification and characterization of the recombinant fkbp-52 (rfkbp-52) was facilitated by incorporating a histidine 6-mer domain at its n terminus. the rfkbp-52 was highly purified on a n(i)2+ affinity resin with an estimated recovery of 10 mg of pure protein from 1 liter of sf9 cell culture. ...	1993	8341706
cdna cloning and expression of a soluble epoxide hydrolase from human liver.	we report the cloning and expression of a cdna that encodes a soluble epoxide hydrolase from human liver. the 2101-base clone predicts a 554-residue protein (m(r) 62,640) with an apparently imperfect peroxisomal targeting signal of ser-lys-met at the carboxy terminus. the cdna was expressed in the baculovirus system in the spodoptera frugiperda 21 cell line. the recombinant protein was similar to soluble epoxide hydrolase isolated from human liver in terms of molecular weight, hydrolytic activit ...	1993	8342951
5-lipoxygenase-activating protein stimulates the utilization of arachidonic acid by 5-lipoxygenase.	5-lipoxygenase (5-lo) and its activating protein (flap) are both required for cellular leukotriene (lt) synthesis, with 5-lo catalyzing both the synthesis of (5s)-5-hydroperoxy-6,8,11,14-eicosatetraenoic acid (5-hpete) from arachidonic acid and the subsequent synthesis of lta4 from 5-hpete. we have previously expressed both human 5-lo and human flap to high levels in spodoptera frugiperda (sf9) insect cells, using recombinant baculoviruses. to study the mechanism by which flap activates 5-lo, we ...	1993	8344271
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
expression and characterization of biologically active human hepatocyte growth factor (hgf) by insect cells infected with hgf-recombinant baculovirus.	a cdna containing the entire coding sequence of human hepatocyte growth factor (hgf) [also known as scatter factor (sf)] was inserted into the genome of autographa california nuclear polyhedrosis virus (baculovirus) adjacent to the polyhedrin promoter by homologous recombination. insect cells (spodoptera frugiperda) infected with the recombinant virus secrete relatively high levels (3-8 mg/l) of biologically active hgf into the culture medium. the recombinant hgf induces pronounced morphological ...	1993	8347597
the presence of udp-n-acetylglucosamine:alpha-3-d-mannoside beta 1,2-n-acetylglucosaminyltransferase i activity in spodoptera frugiperda cells (iplb-sf-21ae) and its enhancement as a result of baculovirus infection.	a golgi preparation from spodoptera frugiperda (iplb-sf-21ae) cells was incubated in the presence of the mannosidase ii inhibitor, swainsonine, with the oligosaccharide, m(alpha 1,3)[[m(alpha 1,3)[m(alpha 1,6)]m(alpha 1,6)]] m(beta 1,4)gn(beta 1,4)gn (m5gn2), the preferred substrate for the enzyme, udp-n-acetylglucosamine:alpha-3-d-mannoside beta 1,2-n-acetylglucosaminyltransferase i (gn-ti). this resulted in formation of the product, gn(beta 1,2)m(alpha 1,3)[[m(alpha 1,3)[m(alpha 1,6)]m(alpha 1 ...	1993	8349674
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
expression of human il-2 receptor alpha- and beta-chains using the baculovirus expression system.	the genes encoding the alpha- and beta-chains of the human interleukin-2 receptor were expressed in lepidopteran insect cells using the baculovirus expression vector system. the corresponding genes were inserted under the polyhedrin promoter of the autographa california nuclear polyhedrosis virus and expressed in the spodoptera frugiperda insect cell line during viral infection. the recombinant receptor proteins were identified in the insect cell lysates by using protein dot blot and elisa techn ...	1993	8356402
steroid requirement for androgen receptor dimerization and dna binding. modulation by intramolecular interactions between the nh2-terminal and steroid-binding domains.	infection of spodoptera frugiperda sf9 insect cells with recombinant human androgen receptor (ar) baculovirus results in expression of a 118-kda phosphoprotein that displays high affinity androgen binding and androgen-dependent targeting to the nucleus. using the dna mobility shift assay, specific in vitro binding of full-length ar to androgen response element dna (are) requires intracellular hormone exposure. the ability of a variety of steroids to induce are binding paralleled their transcript ...	1993	8360187
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
baculovirus-mediated expression of human 65 kda and 67 kda glutamic acid decarboxylases in sf9 insect cells and their relevance in diagnosis of insulin-dependent diabetes mellitus.	cdnas coding for the full-length human 65 and 67 kda glutamic acid decarboxylases (gad65 and gad67) were amplified from pancreas and hippocampus cdna libraries by polymerase chain reaction, respectively. both cdnas were inserted into a baculovirus vector which mediated highly efficient expression of the human gad65 and gad67 with histidine-hexapeptides as affinity ligands at their c-termini in spodoptera frugiperda (sf9) cells. the recombinant gad proteins were purified to homogeneity by affinit ...	1993	8370667
sequence and temporal appearance of the early transcribed baculovirus gene he65.	we have identified the early transcribed he65 gene by screening a cdna library from polyadenylated rna which was isolated at 1 h after infection of spodoptera frugiperda cells with autographa californica nuclear polyhedrosis virus (acnpv). nucleotide sequencing analysis of the he65-specific cdna clone reveals one open reading frame of 1,662 nucleotides from which a protein of 65 kda in size can be predicted. the he65 gene is located downstream of the late transcribed p80 gene and upstream of the ...	1993	8371346
characterization of the replication of plasmids containing hr sequences in baculovirus-infected spodoptera frugiperda cells.	the replication of a series of plasmids, each containing one or more of 6 homologous regions (hrs) from the autographa californica multinucleocapsid nuclear polyhedrosis virus (acmnpv) genome, was investigated in acmnpv-infected spodoptera frugiperda cells. although a construct containing hr2 showed the most efficient replication, the sequences flanking the hr region appeared to influence the levels of replication. plasmids with non-hr-containing acmnpv inserts showed greatly reduced replication ...	1993	8372444
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
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
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
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
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
expression of rat liver phenylalanine hydroxylase in insect cells and site-directed mutagenesis of putative non-heme iron-binding sites.	rat liver phenylalanine hydroxylase was expressed in both escherichia coli and the spodoptera frugiperda insect cell line, sf9. recombinant enzyme from e. coli was inactive and contained less than 0.1 iron atom/subunit. in contrast, recombinant enzyme expressed in sf9 cells using a baculovirus vector was active and identical in several properties to phenylalanine hydroxylase from rat liver: the km for 6-methyltetrahydropterin was 39 microm (compared with 35 microm for the rat liver enzyme), 1 at ...	1993	8385134
development of baculovirus triple and quadruple expression vectors: co-expression of three or four bluetongue virus proteins and the synthesis of bluetongue virus-like particles in insect cells.	baculovirus multiple gene transfer vectors pacab3 and pacab4 have been developed to facilitate the insertion of three or four foreign genes respectively into the autographa californica nuclear polyhedrosis virus (acnpv) genome by a single co-transfection experiment. the pacab3 vector contains a polyhedrin promoter and two p10 promoters on either side of the polyhedrin promoter but in opposite orientations. the pacab4 vector has an additional polyhedrin promoter in opposite orientation to the fir ...	1993	8385313
baculovirus-expressed herpes simplex virus type 2 glycoprotein d is immunogenic and protective against lethal hsv challenge.	herpes simplex virus type 2 glycoprotein d (gd2) was cloned and expressed in the baculovirus-spodoptera frugiperda system. milligram quantities of glycoprotein were recovered from suspension culture and subjected to purification by ion-exchange and immunoaffinity chromatography. the resultant purified gd existed as a homogeneous 57,500 mw monomeric species demonstrating reactivity with anti-gd monoclonal antibodies including those directed at a non-sequential neutralizing epitope of gd. immuniza ...	1993	8385842
human beta 2-adrenergic receptor produced in stably transformed insect cells is functionally coupled via endogenous gtp-binding protein to adenylyl cyclase.	spodoptera frugiperda insect cells (sf9) containing the stably integrated human beta 2-adrenergic receptor gene under the control of the baculovirus ie1 promoter expressed up to 350,000 human receptors/cell. the number of receptors did not change with cell density or age of culture. the adrenergic receptors overexpressed in the insect cells were functional with respect to their ligand binding and signalling properties. coupling of the receptors to endogenous gtp-binding proteins is demonstrated ...	1993	8386637
possible role of phosphorylation in the function of chicken myod1.	chicken myod1 (cmd1), an equivalent to the mouse myod1, expressed in chicken skeletal muscle (lin, z.-y., dechesne, c. a., eldridge, j., and paterson, b. m. (1989) genes & dev. 3, 986-996), was produced in spodoptera frugiperda (sf9) cells by baculovirus expression vector and purified to almost homogeneity. this cmd1 was directly demonstrated to be a phosphoprotein by a 32p-labeling experiment. phosphoamino acid analysis revealed that only serine residue was phosphorylated. phosphoamino acid of ...	1993	8389359
nucleotide sequence of the ubiquitin-39k gene region from the orgyia pseudotsugata multinucleocapsid nuclear polyhedrosis virus genome.	the region of the orgyia pseudotsugata multinucleocapsid nuclear polyhedrosis virus (opmnpv) genome containing the ubiquitin and the nuclear matrix-associated protein (39k) genes was sequenced. the first 77 amino acids of the opmnpv ubiquitin open reading frame (orf) showed 84.4% and 79% amino acid sequence identity to autographa californica multinucleocapsid nuclear polyhedrosis virus (acmnpv) and the spodoptera frugiperda insect ubiquitin, respectively. the predicted opmnpv ubiquitin protein c ...	1993	8389803
recovery and characterization of a rotavirus outer capsid protein expressed in a recombinant insect cell system.	recombinant osu vp4 protein, and outer capsid antigen of porcine rotavirus, was purified to a high level from the spent broth of baculovirus-infected spodoptera frugiperda insect cells. initial clarification of the broth with a 0-60% ammonium sulfate cut retained 93% of the total vp4. q-sepharose ion exchange chromatography performed at ph 6.5 yielded 67% of the initial amount of vp4 in the pooled fractions, with more than four times the purity of the original sample. gel filtration chromatograp ...	1993	8390880
expression of the bovine viral diarrhoea virus osloss p80 protein: its use as elisa antigen for cattle serum antibody detection.	the putative gene encoding the cytopathic bovine viral diarrhoea virus (bvdv) osloss strain p80 protein was amplified by pcr and inserted into a t7 promoter-based vector for expression in escherichia coli. bacterial expression led to cytoplasmic insoluble inclusion bodies which were denatured by urea treatment and renatured by dialysis. rabbit antisera were raised against this p80 recombinant antigen and assayed for the immunoprecipitation of either p120 or p80 protein from cytopathic or non-cyt ...	1993	8393083
synthesis of immunogenic hepatitis a virus particles by recombinant baculoviruses.	recombinant baculoviruses were constructed which contained the hepatitis a virus (hav) open reading frame (orf) under the control of the polyhedrin promoter. northern blot analysis with an hav-specific oligonucleotide probe demonstrated a single transcript large enough to include the hav orf in spodoptera frugiperda cells infected with these recombinants. immunoblots revealed a 220 kda protein representing the hav polyprotein. in addition, proteins which co-migrated with hav capsid proteins, and ...	1993	8393604
the multifunctional folic acid synthesis fas gene of pneumocystis carinii encodes dihydroneopterin aldolase, hydroxymethyldihydropterin pyrophosphokinase and dihydropteroate synthase.	the nucleotide sequence of a folic acid synthesis (fas) gene from pneumocystis carinii contains an open reading frame (orf) that predicts a protein of 740 amino acids with an m(r) of 83,979. a recombinant baculovirus was constructed which directed expression of the predicted fas740 polypeptide in cultured spodoptera frugiperda (sf9) insect cells. the overexpressed 'full-length' protein migrated anomalously in sodium dodecyl sulfate/polyacrylamide gels, with an apparent molecular mass of 71.5 kda ...	1993	8397083
purification of the bovine nicotinic acetylcholine receptor alpha-subunit expressed in baculovirus-infected insect cells.	the bovine skeletal muscle nicotinic acetylcholine receptor alpha-subunit was produced in insect, spodoptera frugiperda (sf9), cells by infection with a recombinant baculovirus. the expressed alpha-subunit protein could not be solubilized efficiently with triton x-100 or sodium cholate, but could be solubilized efficiently with zwittergent 3-14, sodium dodecyl sulfate or tris(hydroxymethyl)aminomethane dodecylsulfate. after solubilization of the alpha-subunit protein with zwittergent 3-14 from t ...	1993	8407866
expression of feline immunodeficiency virus gag and env precursor proteins in spodoptera frugiperda cells and their use in immunodiagnosis.	the gag and env genes of the feline immunodeficiency virus strain ut113 were cloned into a baculovirus transfer vector. the recombinant plasmids were used to create recombinant baculoviruses that expressed either the gag or the env precursor protein in insect cells (sf9 cells). leader sequence cleavage occurred in sf9 cells expressing the envelope precursor, but further processing was not observed. crude lysates of insect cells infected with the wild-type baculovirus or with the recombinant viru ...	1993	8408554
post-translational processing of barley beta-glucan endohydrolases in the baculovirus-insect cell expression system.	two cdnas encoding barley (1-->3,1-->4)-beta-glucanase (ec 3.2.1.73) isoenzymes ei and eii have been expressed in spodoptera frugiperda (sf9) cell cultures using the baculovirus acnpv vector. modifications to both the 5' and 3' ends of the cdnas were required before satisfactory levels of expression were obtained. the modified cdnas directed high levels of (1-->3,1-->4)-beta-glucanase expression in the sf9 insect cell cultures, with yields of approximately 10 mg/liter of isoenzyme ei (expei) and ...	1993	8422276
immunocytochemical characterization of p24, a baculovirus capsid-associated protein.	an open reading frame (orf 1) located upstream of the polyhedron envelope protein gene in the orgyia pseudotsugata multinucleocapsid nuclear polyhedrosis virus (opmnpv) genome was cloned in-frame into a trpe bacterial expression vector. the fusion protein produced by this construct was used for the preparation of a monospecific antiserum. western blot analysis of extracts from opmnpv-infected lymantria dispar cells and autographa californica npv (acmnpv)-infected spodoptera frugiperda cells dete ...	1993	8423444
antibody response to the m2 protein of influenza a virus expressed in insect cells.	a recombinant baculovirus expressing the m2 protein from influenza a/ann arbor/6/60 (h2n2) virus (aa60 virus) was constructed. the expressed m2 protein was recognized by a monoclonal antibody specific for the m2 protein and comigrated with the m2 protein from cells infected with aa60 virus on sds-polyacrylamide gels. immunofluorescence studies indicated that the expressed m2 protein was present on the surface of spodoptera frugiperda (sf9) cells infected with the recombinant baculovirus. immunoa ...	1993	8423445
immunoreactivity and protective effects in mice of a recombinant dengue 2 tonga virus ns1 protein produced in a baculovirus expression system.	the double-stranded replicative form of dengue 2 virus (den-2) rna (tonga strain) was used as a substrate to produce dna clones of the ns1-ns2a genes via reverse transcriptase synthesis of full length cdna followed by polymerase chain reaction amplification of the ns1-ns2a region. products were cloned into ptz18r for sequencing and into baculovirus for expression studies. the deduced amino acid sequence of the ns1-ns2a was almost identical to that of the s1 attenuated strain of den-2 puerto rico ...	1993	8423452
expression of bombyx mori cytoplasmic polyhedrosis virus polyhedrin in insect cells by using a baculovirus expression vector, and its assembly into polyhedra.	a cdna encoding the cytoplasmic polyhedrin of bombyx mori cytoplasmic polyhedrosis virus (bmcpv) strain h was introduced into an improved baculovirus expression vector which can be utilized to express foreign genes in the spodoptera frugiperda cell line iplb-sf-21ae (sf21 cells) and the b. mori cell line bmn. a recombinant virus produced large, cubic inclusion body-like structures in infected sf21 and bmn cells. western blot analysis showed that these structures were bmcpv polyhedra. this result ...	1993	8423453
expression of biologically active human factor viii using a baculovirus vector.	factor viii is a complex, plasma glycoprotein involved in the process of blood coagulation. production of the recombinant molecule has largely been confined to mammalian cell systems which have, in general, proven to be inefficient producers of factor viii. the use of a baculovirus expression system may provide increased levels of this glycoprotein, although it is not certain that insect cell-derived factor viii will be biologically active. the n-linked glycosylation patterns in insect cells, un ...	1993	8427596
production of recombinant androgen receptor in a heterologous expression system.	to facilitate detailed studies of androgen receptor, we have produced a full-length receptor protein and some of its deletion mutants in spodoptera frugiperda (sf9) insect cells, using the baculovirus expression system. recombinant baculovirus dna-infected sf9 cells expressed these proteins in very high quantities, which represented as much as 30-40% of total insect cell protein at 72 h after infection. only < 1% of the recombinant protein was soluble in low-salt buffers; the majority formed ele ...	1993	8432026
site-directed mutagenesis and chemical modification of histidine residues on an alpha-class chick liver glutathione s-transferase cl 3-3. histidines are not needed for the activity of the enzyme and diethylpyrocarbonate modifies both histidine and lysine residues.	each chick liver glutathione s-transferase cl 3 subunit contains three histidine residues: his142, his158 and his228. cl 3-3 can be inactivated by treating with diethylpyrocarbonate. the inactivation process is ph dependent and the pka of the modified residue is 6.4. the second-order inhibition rate constant is 741 m-1min-1 at ph 7.0. based on difference-spectrum and kinetic analysis, inactivation coincides with the modification of one histidine residue. however, hydroxylamine treatment of the d ...	1993	8436137
expression of human thromboxane synthase using a baculovirus system.	human thromboxane (tx) synthase (ec 5.3.99.5) was produced by the baculovirus expression system using cdna encoding human tx synthase [(1991) biochem. biophys. res. commun. 78, 1479-1484]. a recombinant baculovirus txs7 was expressed in spodoptera frugiperda sf9 insect cells. the expressed protein was recognized by monoclonal antibody, kon 7 raised against human tx synthase [(1990) blood 76, 80-85]. the recombinant tx synthase catalyzed the conversion of prostaglandin (pg) h2 to txa2 and 12-hydr ...	1993	8436233
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
bovine respiratory syncytial virus nucleocapsid protein expressed in insect cells specifically interacts with the phosphoprotein and the m2 protein.	the coding region of the gene for the nucleocapsid (n) protein of bovine respiratory syncytial virus has been inserted into the genome of autographa californica nuclear polyhedrosis virus (acnpv) using transfer vector pvl 1393. infection of spodoptera frugiperda cells with recombinant virus resulted in the synthesis of high levels of n protein. this protein was indistinguishable from the authentic bovine respiratory syncytial virus (brsv) n protein by sds-gel electrophoresis and immunoprecipitat ...	1993	8438581
5-lipoxygenase-activating protein is an arachidonate binding protein.	5-lipoxygenase-activating protein (flap) is an 18-kda integral membrane protein which is essential for cellular leukotriene (lt) synthesis, and is the target of lt biosynthesis inhibitors. however, the mechanism by which flap activates 5-lo has not been determined. we have expressed high levels of human flap in spodoptera frugiperda (sf9) insect cells infected with recombinant baculovirus, and used this system to demonstrate that flap specifically binds [125i]l-739,059, a novel photoaffinity ana ...	1993	8440384
functional human transcobalamin ii isoproteins are secreted by insect cells using the baculovirus expression system.	transcobalamin ii (tcii) is a cobalamin (cbl, vitamin b12)-binding protein in mammalian plasma that facilitates the cellular uptake of the vitamin. to obtain human tcii in sufficient quantity for analytical studies, the complementary dna (cdna) encoding tcii was inserted into the plasmid pvl 1393, and the baculovirus expressing tcii was obtained by homologous recombination in spodoptera frugiperda (sf9) insect cells by cotransfection with the wildtype virus. under optimized conditions, sf9 cells ...	1993	8443384
an apoptosis-inhibiting baculovirus gene with a zinc finger-like motif.	spodoptera frugiperda sf-21 cells infected with autographa californica nuclear polyhedrosis virus mutants which lack a functional p35 gene undergo apoptosis, a type of programmed cell death. to identify p35-homologous genes in other baculoviruses, a. californica nuclear polyhedrosis virus dna containing a deletion in p35 was cotransfected into sf-21 cells along with genomic dnas from other baculoviruses. one of the viral dnas which were able to rescue wild-type infection was from cydia pomonella ...	1993	8445726
characterization of vaccinia surface antigen expressed by recombinant baculovirus.	the gene encoding the vaccinia surface antigen (s antigen) was inserted into a baculovirus transfer vector and a recombinant virus was isolated. the s antigen was expressed on the surface of spodoptera frugiperda cells (sf cells) infected with the recombinant baculovirus. recombinant proteins were detected in immunoblotting with anti-vaccinia serum and have the apparent molecular weight of 40 and 50-kda. the 50-kda polypeptide was tunicamycin sensitive and was thus glycosylated. the glycosylated ...	1993	8460484
the expression of hemolytically active human complement protein c9 in mammalian, insect, and yeast cells.	the cdna sequence encoding mature human c9 protein and its signal peptide was cloned into three expression vectors for expression in cos-7 (mammalian), spodoptera frugiperda iplb-sf-21ae (insect), and saccharomyces cerevisiae (yeast) cells. in addition, c9 cdna encoding only the mature protein was fused to the yeast invertase leader sequence (suc2) and cloned for expression in yeast. under optimal conditions cos-7 and iplb-sf-21ae cells secreted recombinant c9 (rc9) at concentrations of about 11 ...	1993	8471847
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
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
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
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