| insect cell culture: improved media and methods for initiating attached cell lines from the lepidoptera. | several cell lines from the pupae of the noctuid moth species spodoptera frugiperda, heliothis zea, and trichoplusia ni were isolated on a synthetic medium containing insect hemolymph and turkey serum. these lines were progressively adapted to improved media free of insect hemolymph but containing one or more of the following sera: turkey, chicken, and fetal calf. primary culture tissue disruption was improved by substituting collagenase for trypsin. primary culture survival was improved by cont ... | 1975 | 172433 |
| plaque assay of baculoviruses employing an agarose-nutrient overlay. | four baculoviruses produced visible plaques in the spodoptera frugiperda cell line when cell monolayers were infected and overlayed with a simplified, agarose-nutrient formulation. macroscopic plaques were first detected 4 days postinoculation, and by 10 days plaques ranging from 0.5 to 3 mm in diameter were seen. dose-response experiments indicated that a single particle initiated the formation of a plaque, because a linear response was demonstrated with increased dosage. | 1979 | 429141 |
| the replication and titration of iridescent virus type 22 in spodoptera frugiperda cells. | a plaque assay for iridescent virus type 22 (from simulium sp.) using spodoptera frugiperda cells has been devised, and the kinetics of growth of the virus in this cell line have been determined. the virus particle/p.f.u. ratio was 75 +/- 8, and the p.f.u./tcid50 ratio was 0.56 +/- 0.11. | 1978 | 621497 |
| the effects of inhibitors of nucleic acid and protein synthesis on the replication of spodoptera frugiperda nuclear polyhedrosis virus in cultured spodoptera frugiperda cells. | the effects of inhibitors of nucleic acid and protein synthesis on the replication of spodoptera frugiperda nuclear polyhedrosis virus have been determined. two inhibitors of protein synthesis-cycloheximide and puromycin-were irreversible inhibitors of virus multiplication. three inhibitors of nucleic acid synthesis-actinomycin d, cytosine arabinoside and camptothecin- prevented virus multiplication; only camptothecin was reversible. rifampicin had no effect on virus multiplication. | 1976 | 799627 |
| replication of nuclear polyhedrosis virus in a continuous cell culture of spodoptera frugiperda: microscopy study of the sequence of events of the virus infection. | a microscopy study of the sequence of morphogenic events of spodoptera frugiperda nuclear polyhedrosis virus infection of s. frugiperda cells is presented which orders the sequence of replication and establishes the time scale within which the events occur. the virus entered the cell by 1 h postinfection and was uncoated. the eclipse period was 9 h and the latent period was 12 h. polyhedron formation was detected by 18 h postinfection and continued until the deposition of the polyhedron membrane ... | 1975 | 1107597 |
| purification and characterization of recombinant xenopus poly(a)(+)-binding protein expressed in a baculovirus system. | the poly(a)(+)-binding protein (pabp) is a highly conserved protein that binds to the poly(a)+ tail of mrnas. pabp has been shown to regulate message stability and translational efficiency, yet the mechanisms remain unknown. to facilitate further dissection of the functions of this protein, we have expressed and purified xenopus pabp using a baculovirus expression system. at 48 h after infection of insect spodoptera frugiperda (sf9) cells with recombinant virus, approx. 3% of cell protein was pa ... | 1992 | 1280105 |
| bacterial luciferase produced with rapid-screening baculovirus vectors is a sensitive reporter for infection of insect cells and larvae. | bacterial luciferase, derived from a fusion of the luxa and luxb genes of vibrio harveyi, has been expressed at very high levels in caterpillars and insect cells. the coding sequence for luciferase was inserted into vectors developed in our laboratory which were designed to expedite screening of recombinant virus. these vectors contained the beta-galactosidase indicator gene under control of immediate early (ie1), early (etl), or very late (p10) promoters and a cloning site for inserting the fus ... | 1992 | 1308004 |
| herpes simplex virus type 1 origin-dependent dna replication in insect cells using recombinant baculoviruses. | the minimal set of seven herpes simplex virus type 1 (hsv-1) genes required for viral origin-dependent dna synthesis was previously identified using a transient replication assay in a mammalian cell line permissive for hsv-1 growth. we have constructed recombinant baculoviruses which efficiently express the products of each of these seven genes in infected spodoptera frugiperda (sf) insect cells. when sf cells were transfected with a plasmid containing a functional hsv-1 origin of replication, a ... | 1992 | 1311360 |
| in vitro transactivation of baculovirus early genes by nuclear extracts from autographa californica nuclear polyhedrosis virus-infected spodoptera frugiperda cells. | nuclear extracts, prepared from autographa californica nuclear polyhedrosis virus-infected spodoptera frugiperda cells during a time course of infection, were analyzed for activation of early gene transcription and for late gene transcription. the templates used in the in vitro transcription assays contained promoters for baculovirus genes that have been classified as immediate early, delayed early, and late. the promoters were derived from the baculovirus 39k, p26, gp64, and dna polymerase gene ... | 1992 | 1316463 |
| physical and functional interaction of human cytomegalovirus dna polymerase and its accessory protein (icp36) expressed in insect cells. | expression of the human cytomegalovirus (hcmv) (ad169) dna polymerase gene under the control of the polyhedrin promoter of autographa californica nuclear polyhedrosis virus in spodoptera frugiperda (sf9) cells has provided a source of highly active cmv dna polymerase. in extracts from cmv-infected cells, the cmv dna polymerase is found strongly associated with an additional polypeptide, icp36. this protein has been identified as the cmv homolog of the herpes simplex virus type 1 ul42 gene produc ... | 1992 | 1318399 |
| expression of the nonstructural proteins of sindbis virus in insect cells by a baculovirus vector. | the genome of sindbis virus encodes the polypeptides that are required for the replication and transcription of the virus rna in infected cells. these polypeptides are translated as a polyprotein that is co- and post-translationally cleaved by an autoproteinase to give rise to four polypeptides designated nsp1, nsp2, nsp3 and nsp4. we have initiated a study of the functions of these proteins by expressing them in the autographa californica baculovirus polyhedrin expression system. spodoptera fru ... | 1992 | 1320794 |
| baculovirus expression of pestivirus non-structural proteins. | bovine viral diarrhoea virus (bvdv) belongs to the pestivirus group, a genus within the flaviviridae family. it possesses a positive-sense ssrna genome with a single large open reading frame (orf) encoding about 4000 amino acids. here we report the continuation of our studies of pestivirus protein biogenesis, involving expression from the viral non-structural protein-encoding region. the 3'-terminal 60% of the bvdv orf was cloned into a plasmid transfer vector which was then used to construct a ... | 1992 | 1321220 |
| characterization of the human prolyl 4-hydroxylase tetramer and its multifunctional protein disulfide-isomerase subunit synthesized in a baculovirus expression system. | prolyl 4-hydroxylase (ec 1.14.11.2), an alpha 2 beta 2 tetramer, catalyzes the posttranslational formation of 4-hydroxyproline in collagens. the enzyme can easily be dissociated into its subunits, but all attempts to associate a tetramer from the dissociated subunits in vitro have been unsuccessful. molecular cloning of the catalytically important alpha subunit has identified two types of cdna clone due to mutually exclusive alternative splicing. the beta subunit is a highly unusual multifunctio ... | 1992 | 1323838 |
| site-directed mutagenesis of human protein disulphide isomerase: effect on the assembly, activity and endoplasmic reticulum retention of human prolyl 4-hydroxylase in spodoptera frugiperda insect cells. | protein disulphide isomerase (pdi) is a highly unusual multifunctional polypeptide, identical to the beta-subunit of prolyl 4-hydroxylase. it has two -cys-gly-his-cys- sequences which represent two independently acting catalytic sites of pdi activity. we report here on the expression in baculovirus vectors of various mutant pdi/beta-subunits together with a wild-type alpha-subunit of the human prolyl 4-hydroxylase alpha 2 beta 2 tetramer in spodoptera frugiperda insect cells. when either one or ... | 1992 | 1327760 |
| expression in insect cells and immune reactivity of a 28k tegument protein of human cytomegalovirus. | the gene encoding the highly antigenic 28k (pp28) tegument phosphoprotein of human cytomegalovirus (hcmv) was expressed in insect cells utilizing a recombinant baculovirus. the mature intracellular form of the recombinant-derived pp28 had mobility on sds-polyacrylamide gels similar to that of native pp28 from hcmv strain towne-infected human foreskin fibroblasts (hffs). in vitro labelling of recombinant autographa california nuclear polyhedrosis virus-infected spodoptera frugiperda cells or of h ... | 1992 | 1328492 |
| assembly of functional gabaa receptors in insect cells using baculovirus expression vectors. | we have constructed recombinant baculoviruses containing cdnas encoding either the alpha 1- or the beta 1-subunit of the bovine gabaa receptor. in spodoptera frugiperda (iplb-sf-21) cells infected with recombinant virus expressing either the alpha 1- or beta 1-subunit, or in cells co-infected with both viruses, functional gabaa receptors were detected by whole-cell electrophysiological recordings. the threshold for the responses mediated by the homo-oligomeric channels (alpha- or beta-) was 2-3 ... | 1992 | 1330062 |
| baculovirus-mediated expression of retinoic acid receptor type gamma in cultured insect cells reveals a difference in specific dna-binding behavior with the 1,25-dihydroxyvitamin d3 receptor. | the baculovirus genetic expression system has been used to produce murine retinoic acid receptor (rar) type gamma in spodoptera frugiperda insect cells and manduca sexta insect larvae. a hydroxyapatite binding assay revealed production levels of 300 pmol of unoccupied receptor per mg of protein in insect cells, whereas levels from infected insect larvae were found to average 100 pmol of rar gamma per mg of protein. the cytosolic preparation from infected insect cells exhibited an equilibrium dis ... | 1992 | 1332041 |
| stimulation of thymidine kinase activity in baculovirus infected cells is not due to a virus-coded enzyme. | a polyhedrin-positive recombinant autographa californica nuclear polyhedrosis virus (acnpv) carrying a herpes simplex virus thymidine kinase gene under the control of the synxiv promoter, a fusion of synthetic and linker-modified polyhedrin promoters, has been constructed. when this recombinant baculovirus was used to infect a variant of spodoptera frugiperda cells deficient in thymidine kinase (tk-), a high level of tk activity was detected. these results, in conjunction with the demonstration ... | 1992 | 1333755 |
| cells transfected with human interleukin 6 cdna acquire binding sites for the hepatitis b virus envelope protein. | earlier studies revealed that human interleukin 6 (il-6) contains recognition sites for the hepatitis b virus (hbv) envelope (env) protein, and that il-6 and anti-il-6 antibodies, respectively, inhibited the interaction of cells expressing a receptor for hbv with the pres(21-47) segment of the hbv env protein, encompassing the complementary attachment site for il-6. this suggested that il-6 mediates hbv-cell interactions. we report that: (a) chinese hamster ovary cells transfected with human il- ... | 1992 | 1334115 |
| activating transcription factor-2 dna-binding activity is stimulated by phosphorylation catalyzed by p42 and p54 microtubule-associated protein kinases. | recent studies have detailed the ability of activating transcription factor-2 (atf-2) to mediate adenoviral e1a stimulation of gene expression; however, an endogenous regulator for the transcriptional activity of this protein has not been described. to characterize the regulation of atf-2 activity, we have expressed full-length and truncated peptides corresponding to various regions of the atf-2 protein in bacteria and the baculovirus insect cell system. bacterially expressed truncated (350-505) ... | 1992 | 1337144 |
| cloning and expression of a chick liver glutathione s-transferase cl 3 subunit with the use of a baculovirus expression system. | glutathione s-transferase cl 3 subunits purified from 1-day-old-chick livers were digested with achromobacter proteinase i and the resulting fragments were isolated for amino acid sequence analysis. an oligonucleotide probe was constructed accordingly for cdna library screening. a cdna clone of 1342 bases, pgcl301, encoding a protein of 26209 da was isolated and sequenced. including conservative substitutions, this protein has 75-79% sequence similarity to other alpha family glutathione s-transf ... | 1992 | 1339283 |
| alzheimer beta/a4-amyloid precursor protein: evidence for putative amyloidogenic fragment. | recombinant baculovirus was used to overexpress human alzheimer beta/a4-amyloid precursor protein (app) in spodoptera frugiperda (sf9) cells. lysates of these cells were then analyzed for the presence of carboxyl-terminal fragments of app by an immunoblotting assay using either an antibody against the app cytoplasmic domain (rabbit anti-human 695app645-694) or an antibody against the amino terminus of beta/a4-amyloid (rabbit anti-human 695app586-606). anti-human 695app645-694 identified app holo ... | 1992 | 1345769 |
| detection of protein kinase homologues and viral rna-binding domains utilizing polyclonal antiserum prepared against a baculovirus-expressed ds rna-activated 68,000-da protein kinase. | the p68 protein kinase (referred to as p68 based on its m(r) of 68,000 in human cells) is a serine/threonine kinase induced by interferon treatment and activated by dsrnas. the kinase is under tight controls in virus-infected cells since once activated, it phosphorylates its natural substrate eukaryotic initiation factor 2 (elf-2), leading to potential limitations in functional elf-2 and decreases in protein synthesis initiation. to further delineate the molecular mechanisms underlying kinase re ... | 1992 | 1360180 |
| continuous production of baculovirus in a cascade of insect-cell reactors. | insect cells (spodoptera frugiperda) were cultured in a continuous stirred-tank reactor. the effluent was led to a cascade of another two reactors, each containing half the volume of the cell-growth reactor, where the cells were infected with autographa californica nuclear polyhedrosis virus. for about 10 days production of 10(7) polyhedra (virus particles embedded in a protein capsule) per cm3 was achieved. this short production time compared to previous experiments involving an analogous syste ... | 1990 | 1366563 |
| sparged animal cell bioreactors: mechanism of cell damage and pluronic f-68 protection. | pluronic f-68 is a widely used protective agent in sparged animal cell bioreactors. in this study, the attachment-independent spodoptera frugiperda sf9 insect cell line was used to explore the mechanism of this protective effect and the nature of cell damage in sparged bioreactors. first, bubble incorporation via cavitation or vortexing was induced by increasing the agitation rate in a surface-aerated bioreactor; insect cells were rapidly killed under these conditions of the absence of polyols. ... | 1990 | 1366875 |
| multistage production of autographa californica nuclear polyhedrosis virus in insect cell cultures. | the aim of our study was to establish an efficient system for the in vitro production of the insect pathogenic autographa californica nuclear polyhedrosis virus in a spodoptera frugiperda cell line. we optimized cultivation conditions for cell proliferation as well as for virus replication in a 1.5 litre stirred tank bioreactor. cell and virus propagation were found to be optimal at a constant oxygen tension of 40%. in order to provide sufficient nutrients during virus synthesis filtration and p ... | 1990 | 1366992 |
| microscopic visualization of insect cell-bubble interactions. i: rising bubbles, air-medium interface, and the foam layer. | through the use of microscopic, high-speed video technology, the interactions of two suspended insect cell lines, trichoplusia ni (tn-368) and spodoptera frugiperda (sf-9), with air and oxygen bubbles were studied. events such as cell-bubble attachment, cell-bubble collision, cell transport into the foam layer, and trapping of cells in the foam layer are presented and discussed. based on these observations and those in a companion paper (chalmers, j. j.; bavarian, f. biotechnol. prog. 1991, foll ... | 1991 | 1367169 |
| microscopic visualization of insect cell-bubble interactions. ii: the bubble film and bubble rupture. | in this paper, the second in the series, the use of a microscopic, high-speed video system to study the interactions of two suspended insect cells strains, trichoplusia ni (tn-368) and spodoptera frugiperda (sf-9), with rupturing bubbles is reported. events such as the adsorption of cells onto the bubble film and the mechanism of bubble rupture were observed. on the basis of these observations and the experimental and theoretical work of other researchers on bubble rupture and cell death as a re ... | 1991 | 1367170 |
| dna distribution and respiratory activity of spodoptera frugiperda populations infected with wild-type and recombinant autographa californica nuclear polyhedrosis virus. | spodoptera frugiperda cells were infected with a wild-type autographa californica nuclear polyhedrosis virus and with a recombinant autographa californica nuclear polyhedrosis virus. the recombinant virus was derived from the wild-type virus and produced beta-galactosidase instead of polyhedrin. the changes in cell size, cell growth, viability, dna distribution, and respiratory activity were followed through the time course of the infection. the dna content as measured by flow cytometry of infec ... | 1990 | 1367465 |
| use of a stationary bed reactor and serum-free medium for the production of recombinant proteins in insect cells. | insect cells (spodoptera frugiperda) have been cultured in a stationary bed reactor, packed with a fibrous polyester carrier. when the bioreactor was perfused with serum-supplemented medium, a cell density of 6 x 10(6) cells ml-1 packed carrier was reached. scanning electron microscopy investigations have shown that the insect cells grew along the three-dimensionally oriented fibers of the fibra-cel carrier. after infection of the logarithmically growing cells with a recombinant baculovirus (aut ... | 1991 | 1367637 |
| production of recombinant proteins by baculovirus-infected gypsy moth cells. | an experimental study was undertaken to evaluate alternative insect cell lines to sf9 [from spodoptera frugiperda (fall armyworm)] for the production of recombinant proteins. insect cell lines from two different organisms were considered: iplb-ldeita (ldeita) from lymantria dispar (gypsy moth) and iplb-hvt1 (hvt1) from heliothis virescens (tobacco budworm). both ldeita and hvt1 produced higher total activity levels of recombinant beta-galactosidase in monolayer culture than sf9 after infection w ... | 1991 | 1367995 |
| a sensitive model system for in vivo monitoring of baculovirus gene expression in single infected insect cells. | we have developed a fast and sensitive system for the in vivo analysis of gene expression in baculovirus infected lepidopteran insect cells. a recombinant baculovirus containing a luciferase gene from the click beetle, pyrophorus plagiophthalamus, under transcriptional regulation of the polyhedrin gene promoter of autographa californica nuclear polyhedrosis virus (acnpv) was used to infect a spodoptera frugiperda cell line. recombinant luciferase could be monitored by luminometry in real-time wi ... | 1992 | 1368236 |
| optimization of protein-production by the baculovirus expression vector system in shake flasks. | shake flasks were successfully employed for the cultivation of spodoptera frugiperda (sf-9) insect cells and for the production of beta-galactosidase, a recombinant model protein, utilizing the baculovirus expression vector system. the culture doubling time and maximal cell density were 20 h and 5 x 10(6) cells/ml respectively. the optimal liquid volumes for flasks rotating at 100 rpm were 25-40% of the flask total volume. enzyme production (about 600 mg/l) was best at a multiplicity of infectio ... | 1992 | 1368502 |
| replication of a nuclear polyhedrosis virus in a continuous cell line of spodoptera frugiperda: partial characterization of the viral dna, comparative dna-dna hybridization, and patterns of dna synthesis. | | 1978 | 351928 |
| the establishment of two cell lines from the insect spodoptera frugiperda (lepidoptera; noctuidae). | the history and characteristics of two cells lines developed from primary explants of pupal tissue from the insect, spodoptera frugiperda (j.e. smith), are described. one cell line, iplb-sf 21, was developed with hemolymph-supplemented medium and has been maintained continuously on the medium. the second cell line, iplb-sf-1254, was developed with a medium containing a combination of vertebrate sera plus hemolymph and was adapted to hemolyphn-free medium at the 6th passage. the iplb-sf-21 cell l ... | 1977 | 68913 |
| characteristics of the non-occluded form of a nuclear polyhedrosis virus. | non-occluded virions of a nuclear polyhedrosis virus of the alfalfa looper, autographa californica, found in the medium of cell cultures of infected fall armyworm, spodopter frugiperda, and in the hemolymph of infected s. frugiperda larvae were partially characterized by biological, chemical and physical methods. also, the rate of appearance of the virions was studied in cell culture and the host insect to determine maximum virion production. virions obtained from both sources were heat-sensitiv ... | 1975 | 343 |
| effect of glycosylation on properties of soluble interferon gamma receptors produced in prokaryotic and eukaryotic expression systems. | we investigated the influence of glycosylation on solubility, chromatographic behavior and resistance to heat- and chaotrope-dependent denaturation and proteolytic digestion of three recombinant human interferon gamma receptors produced in escherichia coli, spodoptera frugiperda and chinese hamster ovary cells. the proteins produced in the eukaryotic expression systems were glycosylated, carrying different, heterogeneous carbohydrate moieties. they were assayed fully glycosylated and after remov ... | 1992 | 1368793 |
| baculovirus expression of alkaline phosphatase as a reporter gene for evaluation of production, glycosylation and secretion. | we have devised a simple and efficient baculovirus expression vector system to evaluate insect tissue culture cells for their capacity to express, glycosylate and secrete foreign proteins. a truncated placental alkaline phosphatase (seap) gene was inserted into the autographa californica nuclear polyhedrosis virus (acmnpv) genome under the transcriptional control of the polyhedrin gene promoter. production levels, glycosylation, and secretion of the recombinant protein were examined in trichoplu ... | 1992 | 1368794 |
| trypanosoma cruzi flagellar repetitive antigen expression by recombinant baculovirus: towards an improved diagnostic reagent for chagas' disease. | we constructed a recombinant baculovirus that expressed part of a trypanosoma cruzi flagellar repetitive antigen (fra). both cell- associated and secreted forms of recombinant fra were detected in cultures of virus-infected spodoptera frugiperda (sf9) cells. these forms show a complex pattern after polyacrylamide gel electrophoresis and western blot analysis using either an anti-fra rabbit serum or human chagasic sera. competitive western-blot experiments revealed that all bands react with the s ... | 1992 | 1369025 |
| recombinant beta-galactosidase production in serum-free medium by insect cells in a 14-l airlift bioreactor. | spodoptera frugiperda (sf9) insect cells were successfully cultured in serum-free medium in a 14-l airlift bioreactor. cell densities as high as 1 x 10(7) cells/ml were achieved with specific growth rates of approximately 0.0286 h-1 (doubling time of 24 h). this system was also used to demonstrate the expression of a reported gene, beta-galactosidase (beta-gal), when cells were infected with a recombinant baculovirus. approximately 0.33 mg of beta-gal/ml (i.e., 104,000 units/ml) of medium were o ... | 1992 | 1369040 |
| recombinant protein production in insect cell cultures infected with a temperature-sensitive baculovirus. | spodoptera frugiperda (iplb-sf-21) insect cells were grown in shake-flasks and infected with a temperature-sensitive baculovirus to express the gene of chloramphenicol acetyl transferase (cat) in serum-free medium (sf-900) and two serum-supplemented media (ipl-41 and grace's). in temperature-shift experiments (cell growth at 33 degrees c followed by virus replication at 27 degrees c 3-4 days later), virus and cat production were much poorer in the serum-free medium than in serum-supplemented med ... | 1992 | 1369167 |
| screening of insect cell lines for the production of recombinant proteins and infectious virus in the baculovirus expression system. | eight cell lines derived from the insects spodoptera frugiperda, trichoplusia ni, mamestra brassicae, and estigmene acrea were evaluated for recombinant beta-galactosidase and infectious virus production following infection with the baculovirus autographa californica multiple nuclear polyhedrosis virus (acmnpv). production was assessed on a specific (per cell and per microgram of uninfected cellular protein) and on a volumetric (per milliliter) basis. cell density was found to be an important fa ... | 1992 | 1369220 |
| effects of oxygen on recombinant protein production by suspension cultures of spodoptera frugiperda (sf-9) insect cells. | spodoptera frugiperda (sf-9) insect cells have been grown in serum-free medium in 250-ml spinner flasks. the maximum cell density obtained in these cultures was dependent on the aeration rate of the culture. similar yields of uninfected cells were obtained when cultures were stirred in spinner flasks at 80 rev min-1 and in a 4-1 stirred-tank bioreactor and the dissolved oxygen in the bioreactor was controlled at 20% of air saturation. cells were infected with a recombinant baculovirus at differe ... | 1992 | 1369406 |
| protective effect of methylcellulose and other polymers on insect cells subjected to laminar shear stress. | the relative sensitivity of two insect cell lines to laminar shear stress was determined, and the protective effect of polymers added to the growth media of two insect cell lines, trichoplusia ni (tn-368) and spodoptera frugiperda (sf-9), was evaluated. tn-368 and sf-9 cells were found to be equally sensitive to laminar shear stress. methylcellulose [0.5% (w/v) dow e4m methocel] and dextran [4.5% (w/v)] increased the resistance of suspended cells to lysis due to laminar shear stress by factors o ... | 1990 | 1370016 |
| bovine rotavirus segment 5 protein expressed in the baculovirus system interacts with zinc and rna. | the cdna sequence of genomic segment 5 of bovine rotavirus (rf strain) has been inserted into baculovirus transfer vectors, downstream of the polyhedrin promotor. recombinant baculoviruses containing gene 5 were selected and the protein was expressed to high yields in spodoptera frugiperda cells. the recombinant protein was inoculated into rabbits and mice to produce specific hyperimmune antisera. the polyclonal antiserum reacted with a protein in rotavirus-infected ma104 cells and with a protei ... | 1992 | 1379624 |
| presentation of hepatitis b virus pres2 epitope on bluetongue virus core-like particles. | a chimeric protein containing most of the hepatitis b virus pres2 region (amino acid residues 1-48) upstream to, and colinear with the amino-terminus of bluetongue virus vp7 protein (pres2-vp7) was expressed by a recombinant autographa californica nuclear polyhedrosis virus (acnpv). the chimeric protein formed btv core-like particles (clps) in spodoptera frugiperda cells only when the cells were coinfected with this recombinant virus and a recombinant baculovirus that expresses unmodified vp7 an ... | 1992 | 1381538 |
| potential role of monocyte chemoattractant protein 1/je in monocyte/macrophage-dependent iga immune complex alveolitis in the rat. | we have examined the role of monocyte chemoattractant protein 1 (mcp 1) in the pathogenesis of monocyte/macrophage-dependent iga immune complex alveolitis in the rat. rat mcp 1 was cloned and expressed in order to facilitate analysis of its function in rat models of human disease. a cdna library was constructed from rat pulmonary artery endothelial cells stimulated with tnf-alpha. the cdna library was screened with synthetic oligonucleotide probes based on the recently published rat mcp 1 cdna s ... | 1992 | 1387671 |
| expression, purification, and functional characterization of adenovirus 5 and 12 e1a proteins produced in insect cells. | the 12 s and 13 s e1a cdnas from both the adenovirus (ad) nononcogenic type 5 and the oncogenic type 12 were overexpressed in an insect cell/baculovirus system. upon infection of spodoptera frugiperda cells, the production of e1a proteins reached a level of about 15 micrograms/10(6) cells. the e1a proteins are highly soluble and apparently are processed authentically. they are readily recognized by various antibodies and display phosphorylation patterns similar to those of e1a proteins synthesiz ... | 1992 | 1387752 |
| purification and characterization of a low m(r) gtp-binding protein, ram p25, expressed by baculovirus expression system. | the ram gene was isolated from rat megakaryocyte cdna library with an oligonucleotide probe which is specific for a low m(r) gtp-binding proteins c25kg purified from human platelets. its gene product (ram p25) is a monomeric 25-kda guanine nucleotide-binding protein. the protein was expressed by using baculovirus transfer vector, pacym1, which allowed the production at a high level of soluble recombinant ram p25 in spodoptera frugiperda (sf9) cells under the control of polyhedrin promoter. the e ... | 1992 | 1390922 |
| characterization of rabies virus glycoprotein expressed by recombinant baculovirus. | a cdna of the glycoprotein (g protein) gene of rabies virus nishigahara strain was cloned and inserted into a baculovirus genome under the control of the polyhedrin promoter. infection of spodoptera frugiperda cells with this recombinant virus produced a large quantity of new protein instead of the parental polyhedrin protein. by immunofluorescent and immunoblotting analyses, the recombinant protein was antigenically similar to the authentic g protein. its molecular mass estimated by sodium dode ... | 1992 | 1413990 |
| purification of pancreatic cholesterol esterase expressed in recombinant baculovirus-infected sf9 cells. | a cdna clone encoding the entire coding sequence of rat pancreatic cholesterol esterase (bile salt-stimulated lipase) was subcloned into the baculovirus transfer vector pvl1392 and used to co-transfect spodoptera frugiperda (sf9) insect cells with wild-type autographa californica nuclear polyhedrosis virus (acnpv) dna. two recombinant proteins (m(r) 74 kda and 64 kda) reactive with anti-cholesterol esterase igg were produced and secreted by the infected sf9 cells in large quantities in a time-de ... | 1992 | 1422214 |
| high-level production of human alpha- and beta-globins in insect cells. | high-level production of human alpha- and beta-globins in cultured spodoptera frugiperda (sf-9) cells infected with recombinant baculoviruses is described. the expressed globins are produced to 70-140 mg protein/liter of cell culture or 5-10% of the total cellular protein. two recombinant baculoviruses for alpha-globin, h alpha and h beta alpha, differ in their construction in that the 5'-untranslated region of the beta-globin gene is inserted 5' to the alpha-globin mrna coding region in h beta ... | 1992 | 1422217 |
| production, purification, and characterization of human recombinant il-8 from the eucaryotic vector expression system baculovirus. | the cdna for the human chemotactic interleukin, il-8, was subcloned from a bacterial source into the eucaryotic vector expression system baculovirus. recombinant human il-8 (rhil-8) was synthesized and secreted from sf9 cells derived from spodoptera frugiperda following infection of a recombinant virus harboring the full-length il-8 structural gene. infected sf9 cells produced rhil-8 in a range from 0.5 to 2.0 mg of rhil-8/liter of postinfection cell culture media. the recombinant interleukin wa ... | 1992 | 1422225 |
| recombinant laminin b1 chains exhibit intact short-arm domains but do not form oligomeric molecules. | the human laminin b1 chain has been produced in the baculovirus expression system in sufficient amounts for biochemical and functional studies. a full-length cdna, which was constructed of four partially overlapping clones and verified by in vitro transcription and translation to be functional, was cloned into the transfer vector pvl1392 behind the polyhedrin promoter. the recombinant construct was incorporated by in vivo homologous recombination into the genome of the wild-type baculovirus, aut ... | 1992 | 1425664 |
| expression of cauliflower mosaic virus orf ii in a baculovirus system. | the cauliflower mosaic virus orf ii encoding the aphid transmission factor (atf) was mutagenized to introduce a bamhi restriction site upstream from the initiation codon and then cloned into an eukaryotic viral expression vector (autographa californica nuclear polyhedrosis virus). all recombinant viruses tested in spodoptera frugiperda (sf21) cells expressed a protein of about 18 kd which comigrated in page with atf from infected plants. western blotting using an oligopeptide antiserum to atf co ... | 1992 | 1428752 |
| the identification and characterization of a gdp-dissociation inhibitor (gdi) for the cdc42hs protein. | the ras-related protein, cdc42hs, is a 22-kda gtp-binding protein which is the human homolog of a saccharomyces cerevisiae yeast-cell-division cycle protein. in attempting to isolate and biochemically characterize mammalian proteins capable of regulating various activities of cdc42hs, we have identified an activity in bovine brain cytosol which effectively inhibits the dissociation of [3h]gdp from the platelet- or the spodoptera frugiperda-expressed cdc42hs protein. the purification of this acti ... | 1992 | 1429634 |
| sequence evolution in mitochondrial ribosomal and nd-1 genes in lepidoptera: implications for phylogenetic analyses. | a 2,256-bp sequence of the mitochondrial genome of a lepidopteran (spodoptera frugiperda) contains trnas for valine and leucine, the 16s rrna, and three-quarters of the nd-1 presumptive protein-coding gene. a 64-bp stretch of unknown function was located between the rrna and leucine trna. sequence divergence in the 16s rrna obtained from alignment with published insect sequences is consistent with phylogenetic hypotheses, in that diptera and lepidoptera are more closely related to each other (24 ... | 1992 | 1435234 |
| expression and secretion of japanese encephalitis virus nonstructural protein ns1 by insect cells using a recombinant baculovirus. | the nonstructural protein ns1 of japanese encephalitis virus (jev) was expressed at a high level under the control of the polyhedrin promoter in spodoptera frugiperda (sf9) insect cells using a recombinant baculovirus. recombinant ns1 was designed to contain its natural signal sequence at its n-terminus and no c-terminal hydrophobic domain that could act as a membrane anchor. this recombinant protein exhibited similar size to native ns1 expressed in aedes albopictus (c6/36) insect cells infected ... | 1992 | 1448926 |
| studies on processing, particle formation, and immunogenicity of the hiv-1 gag gene product: a possible component of a hiv vaccine. | antigens in a particulate conformation were shown to be highly immunogenic in mammals. for this reason, the particle forming capacity of derivatives of the hiv-1 group specific core antigen p55 gag was assayed and compared dependent on various expression systems: recombinant bacteria, vaccinia- and baculoviruses were established encoding the entire core protein p55 either in its authentic sequence or lacking the myristylation consensus signal. moreover, p55 gag was expressed in combination with ... | 1992 | 1456888 |
| analysis of soluble human and mouse interferon-gamma receptors expressed in eukaryotic cells. | the extracellular domains of the human and mouse interferon-gamma receptors were produced in insect spodoptera frugiperda cells infected with recombinant baculoviruses and in mammalian chinese-hamster-ovary cells. the receptors expressed in both systems are secreted into the culture medium. their signal peptides are cleaved off and the proteins show heterogeneity in glycosylation which, however, does not affect the capacity to bind interferon gamma or specific antibodies. the soluble mouse recep ... | 1992 | 1459135 |
| studies on the interaction of alpha subunits of gtp-binding proteins with beta gamma dimers. | the interaction of several preparations of purified beta gamma dimers with two types of guanosine-nucleotide-binding-regulatory-(g)-protein alpha subunits, a recombinant bv alpha i3, made in sf9 spodoptera frugiperda cells by the baculovirus (bv) expression system, and alpha s, either purified from human erythrocyte gs-type gtp-binding protein, and activated by naf/alcl3, or unpurified as found in a natural membrane, were studied. the beta gamma dimers used were from bovine rod outer segments (r ... | 1992 | 1459143 |
| distinct signals in human immunodeficiency virus type 1 pr55 necessary for rna binding and particle formation. | the human immunodeficiency virus type 1 (hiv-1) gag gene product pr55 self-assembles to form virus-like particles when expressed in spodoptera frugiperda cells using recombinant baculoviruses. the particles resemble immature hiv and are released from the infected cell into the culture medium. using this system we have progressively truncated the gag open reading frame from the c terminus and examined each deleted gag protein for its particle-producing capability. we show that deletion of pr6 and ... | 1992 | 1469349 |
| transient expression assay in a baculovirus system using firefly luciferase gene as a reporter. | transient gene expression assays were developed to assess the function of the regulatory sequences of baculoviruses bombyx mori nuclear polyhedrosis virus (bmnpv) and autographa californica nuclear polyhedrosis virus (acnpv) in insect cells of bombyx mori and spodoptera frugiperda, respectively. dna sequences encoding luciferase (luc) of the firefly photinus pyralis was successfully employed in the expression assay as a reporter gene. recombinant plasmids were constructed containing the luc gene ... | 1992 | 1475909 |
| characterization of baculovirus-expressed hemagglutinin and fusion glycoproteins of the attenuated measles virus strain aik-c. | with measles virus cdna of the avirulent vaccine strain aik-c, two cdnas of h or f genes were amplified by the polymerase chain reaction. the amplified cdnas were inserted respectively to the baculovirus transfer vector pacym1 derived from the nuclear polyhedrosis virus of autographa californica (acnpv). after co-transfection of the transfer vectors with acnpv dna to spodoptera frugiperda cells, recombinant baculoviruses were screened by plaque assay, and the viruses containing h-cdna or f-cdna ... | 1992 | 1476005 |
| human preproendothelin-1 is converted into active endothelin-1 by baculovirus-infected insect cells. | to investigate biochemical and biological parameters involved in preproendothelin-1 (preproet-1) maturation we infected spodoptera frugiperda (sf21) cells with a suitable engineered baculovirus vector carrying the cdna encoding the entire human 212 amino acids precursor. culture supernatants were tested by ria using an anti-et-1 serum, et-1-like immunoreactive material (irm) was detected in the infected sf21 cells but not in control, wild-type or mock-infected cells. fractionation of the culture ... | 1992 | 1497664 |
| distinct n-glycan fucosylation potentials of three lepidopteran cell lines. | the fucosyltransferase activities of three insect cell lines, mb-0503 (from mamestra brassicae), bm-n (from bombyx mori) and sf-9 (from spodoptera frugiperda), were investigated and compared with that of honeybee venom glands. cell extracts and venom gland extracts were incubated with gdp-[14c]fucose and glycopeptides isolated from human igg and from bovine fibrin. the labeled oligosaccharide products were released by peptide-n4-(n-acetyl-beta-glucosaminyl)asparagine amidase a, fluorescence mark ... | 1992 | 1499571 |
| site-specific mutagenesis of the 35-kilodalton protein gene encoded by autographa californica nuclear polyhedrosis virus: cell line-specific effects on virus replication. | the gene encoding the 35-kda protein (35k gene) located within the ecori-s genome fragment of autographa californica nuclear polyhedrosis virus (acmnpv) is transcribed early in infection. to examine its function(s) with respect to virus multiplication, we introduced specific mutations of this early gene into the acmnpv genome. in spodoptera frugiperda (sf21) culture, deletion of the 35k gene reduced yields of extracellular, budded virus from 200- to 15,000-fold, depending on input multiplicity. ... | 1992 | 1501287 |
| overexpressed full-length human bcl2 extends the survival of baculovirus-infected sf9 insect cells. | full-length and truncated human bcl2 lacking the entire c-terminal hydrophobic domain have been overexpressed in spodoptera frugiperda insect cells with the baculovirus expression system. immunoblot analysis with bcl2-specific antibodies revealed that both full-length and truncated bcl2 are expressed as multiple immunoreactive species, suggesting posttranslational modifications. the expression of the full-length but not the truncated bcl2 extended the survival of baculovirus-infected cells by pr ... | 1992 | 1502141 |
| a synthetic early promoter from a baculovirus: roles of the tata box and conserved start site cagt sequence in basal levels of transcription. | many baculovirus early genes and insect genes transcribed by rna polymerase ii have a conserved transcription start site sequence (cagt) located downstream of a consensus tata box. to examine the functions and interactions of these two motifs in initiating accurately positioned basal transcription, a 43-nt synthetic promoter was synthesized from the tata box and start site sequences of the gp64 early promoter from the orgyia pseudotsugata multicapsid nuclear polyhedrosis virus (opmnpv). the synt ... | 1992 | 1519359 |
| site-directed mutagenesis and chemical modification of cysteine residues of rat glutathione s-transferase 3-3. | rat liver glutathione s-transferase (gst) 3-3 is composed of two identical subunits, each containing three cysteine residues, cys-86, cys-114 and cys-173. we have shown previously that cys-86 is not involved in the enzymic activity of gst 3-3 [hsieh, huang, chen, lai & tam (1991) biochem, j. 278, 293-297]. at 50 degrees c, iodoacetamide can inactivate the enzyme by modifying cys-86 and cys-114. cys-114 can be protected against iodoacetamide inhibition by s-(dinitrophenyl)glutathione. site-direct ... | 1992 | 1520269 |
| expression of two s-ribonucleases of petunia inflata using baculovirus expression system. | we have previously shown that three petunia inflata s-proteins, products of the multiallelic s-gene of the self-incompatibility system, are ribonucleases. here we report the expression of cdnas for two of these s-proteins using the baculovirus expression system. s2- and s3-proteins were found in both supernatants and lysates of spodoptera frugiperda cells infected with recombinant baculoviruses. both recombinant s-proteins contained glycosylated (25 kd) and nonglycosylated (23 kd) forms. recombi ... | 1992 | 1520311 |
| hepatitis delta antigen expressed by recombinant baculoviruses: comparison of biochemical properties and post-translational modifications between the large and small forms. | hepatitis delta virus (hdv) encodes only one protein, the hepatitis delta antigen (hdag). two forms of hdag, a large (27 kda) and a small (24 kda) one, participate in the various steps of hdv replication. to further understand the properties of hdag, we have constructed recombinant baculoviruses and expressed both forms of the hdag in insect cells. the gene encoding hdag was placed under the control of the polyhedrin promoter of autographa californica nuclear polyhedrosis virus (acnpv) by homolo ... | 1992 | 1529540 |
| expression of the nucleocapsid protein of dugbe virus and antigenic cross-reactions with other nairoviruses. | the small (s) rna segment of dugbe (dug) virus (nairovirus, bunyaviridae) encodes a single protein, the nucleocapsid (n) protein, of m(r) 49.4 kda. cdna derived from the complete coding region for the n protein was cloned into autographa californica nuclear polyhedrosis virus (acnpv) under control of the polyhedrin promoter and used to infect spodoptera frugiperda insect cells. western blotting analysis using monoclonal antibodies demonstrated the production of dug n protein in the infected cell ... | 1992 | 1529645 |
| baculovirus-mediated expression and functional characterization of human nadph-p450 oxidoreductase. | human nadph-p450 oxidoreductase (or) is an intrinsically membrane-bound flavoprotein that serves to transfer electrons from nadph to cytochrome p450. or is also involved in the metabolic activation of chemotherapeutic alkylating agents. the human or cdna was engineered into baculovirus and the recombinant virus was used to infect spodoptera frugiperda (sf9) cells. approximately 3.3% of total protein of infected cells was human or. the enzyme was purified by ion exchange and affinity chromatograp ... | 1992 | 1536559 |
| expression of androgen receptor in insect cells. purification of the receptor and renaturation of its steroid- and dna-binding functions. | a full-length rat androgen receptor cdna was used to produce a recombinant baculovirus (acrar) by homologous recombination. spodoptera frugiperda (sf9) cells infected with this virus expressed a 110-kda polypeptide that amounted up to about one-third of total cell protein. studies with ar antibodies confirmed that this protein was indeed rar. only a minor portion of the recombinant ar was soluble in buffers without ionic detergents, but its complete solubilization was achieved in 6 m guanidine h ... | 1992 | 1537872 |
| high-level synthesis and fate of acetylcholine in baculovirus-infected cells: characterization and purification of recombinant rat choline acetyltransferase. | rat choline acetyltransferase (chat) has been expressed at a high level in spodoptera frugiperda sf9 cells using a baculovirus expression system. a cdna containing the coding sequence for chat was inserted into the transfer vector pacym1 to yield the recombinant vector pacym1/chat. sf9 cells were then coinfected with pacym1/chat and the wild-type autographa californica virus. one recombinant virus particle, containing the cdna for chat, was selected that expressed a protein of 68.5 kda. forty ho ... | 1992 | 1548478 |
| biophysical and biochemical properties of baculovirus-expressed camv p1 protein. | cauliflower mosaic virus (camv) gene i encodes a protein (p1) that has been implicated in the control of virus movement in infected plants. to assist in the characterization of the mechanism of action of p1, gene i has been expressed efficiently in spodoptera frugiperda (sf) cells using recombinant baculovirus. control of the expression of camv gene i by the polyhedrin late promoter in the baculovirus autographa californica nuclear polyhedrosis virus (acnpv) resulted in very high levels of p1 ac ... | 1992 | 1549910 |
| expression of prostaglandin endoperoxide synthase-1 in a baculovirus system. | a cdna coding for ovine prostaglandin endoperoxide (pgh) synthase-1 was used to construct a recombinant baculovirus which was expressed in spodoptera frugiperda (sf9) insect cells. two proteins reactive with anti-pgh synthase antibody were produced. a larger protein (mr = 72,000) coelectrophoresed with native enzyme; a smaller, more abundant protein (mr = 66,000) was unglycosylated enzyme. about 90% of both the immunoreactivity and the cyclooxygenase activity were present in a low speed (10(5) g ... | 1992 | 1567411 |
| expression and transport into mitochondria of bovine cytochrome p-450(scc) in insect cells using the baculovirus expression system. | bovine cytochrome p-450(scc) introduced with the baculovirus host vector system was found to be expressed in spodoptera frugiperda cells. cell fractionation analysis indicated that the p-450(scc) expressed as the precursor form was transported into mitochondria and converted to a mature form. however, this form did not exhibit definite activity for cholesterol side chain cleavage. these findings suggest that most of the p-450(scc) expressed by this system is an inactive protein within mitochondr ... | 1992 | 1567449 |
| synapsin iia: expression in insect cells, purification, and characterization. | synapsin iia belongs to a family of neuron-specific phosphoproteins called synapsins, which are associated with synaptic vesicles in presynaptic nerve terminals. in order to examine the biochemical properties of synapsin iia, and ultimately its physiological function, purified protein is required. since attempts to purify significant quantities of synapsin iia, an isoform of the synapsins, from mammalian brain have proven difficult, we undertook the production of recombinant synapsin iia by util ... | 1992 | 1567872 |
| the nadp-dependent methylenetetrahydrofolate dehydrogenase-methenyltetrahydrofolate cyclohydrolase-formyltetrahydrofolate synthetase is not expressed in spodoptera frugiperda cells. | the insect cell line derived from spodoptera frugiperda (sf9) does not express the activities of the trifunctional nadp-dependent methylenetetrahydrofolate dehydrogenase-methenyltetrahydrofolate cyclohydrolase-formyltetrahydrofolate synthetase. the lack of synthetase activity was confirmed by the inability to incorporate radiolabeled formate into nucleotides. the cells express, instead, a mg2+ and nad-dependent bifunctional methylenetetrahydrofolate dehydrogenase-methenyltetrahydrofolate cyclohy ... | 1992 | 1569082 |
| differential factor binding at the promoter of early baculovirus gene pe38 during viral infection: gata motif is recognized by an insect protein. | regulatory elements interacting with dna-binding proteins have been investigated in the promoter sequence of the early pe38 gene in the autographa californica nuclear polyhedrosis virus (acnpv). a gata motif located 50 nucleotides upstream of the pe38 transcriptional start site is recognized differentially in the course of infection. as demonstrated by footprint and gel mobility shift assays, the gata sequences ttatct are protected by nuclear extracts from uninfected spodoptera frugiperda cells ... | 1992 | 1583720 |
| isopentenoid synthesis in embryonic drosophila cells: prenylated protein profile and prenyl group usage. | it has been established that vertebrates and yeasts modified a unique subset of polypeptides with farnesyl and geranylgeranyl residues. this observation has been extended to drosophila kc cells. [3h]mevalonate was incorporated into 54 kc cell peptides (18-92 kda). as reported for mammalian cells, most of the labeled peptides had molecular weights between 21 and 27 kda. c18 radio-hplc tryptic digest profiles for delipidized, [3h]mevalonate-labeled (a) insect (drosophila and spodoptera frugiperda) ... | 1992 | 1586169 |
| 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 |
| high level expression of dna polymerases from herpesviruses. | the dna polymerase genes of human cytomegalovirus (hcmv) and varicella-zoster virus (vzv) were inserted separately into the polyhedrin gene of autographa californica nuclear polyhedrosis virus (acnpv) by cotransfection of spodoptera frugiperda (sf9) cells with baculovirus transfer vectors carrying the genes and acnpv infectious dna. infection of sf9 cells with the recombinant viruses resulted in expression from the polyhedrin promoter of proteins of the expected mrs. these proteins possessed dna ... | 1991 | 1649906 |
| immunological characterization of the epstein-barr virus phosphoprotein pp58 and deoxyribonuclease expressed in the baculovirus expression system. | the open reading frames of the phosphoprotein pp58 (bmrfi) and the deoxyribonuclease (bglf5) of the epstein-barr-virus (ebv) strain m-aba were cloned in the baculovirus expression vectors pac373 and pac360 and expressed in the spodoptera frugiperda (sf158) insect cells. the recombinant phosphoprotein pp58 expressed in sf158 cells was recognized by the anti-pp58 rabbit anti-sera which were generated by immunizing rabbits with a trpe-bmrfi fusion protein expressed in e. coli. dna-cellulose chromat ... | 1991 | 1650330 |
| overproduction of rat 1,25-dihydroxyvitamin d3 receptor in insect cells using the baculovirus expression system. | the rat 1,25-dihydroxyvitamin d3 [1,25-(oh)2d3] receptor has been expressed at elevated levels in spodoptera frugiperda cells using the baculovirus expression vector system. the recombinant 1,25-(oh)2d3 receptor is full-length, binds 1,25-(oh)2d3, and is recognized by a monoclonal antibody specific for 1,25-(oh)2d3 receptor. densitometric scanning of coomassie brilliant blue-stained sds/polyacrylamide gels indicated a recombinant receptor protein level comprising 5% of the total soluble protein ... | 1991 | 1650474 |
| expression and characterization of poliovirus proteins 3bvpg, 3cpro, and 3dpol in recombinant baculovirus-infected spodoptera frugiperda cells. | as an initial step toward investigating the roles of poliovirus proteins in viral rna replication, a baculovirus expression system was used to produce poliovirus proteins from the p3 region. spodoptera frugiperda (sf9) cells were infected with a recombinant baculovirus, vetl-pov3a*bcd, which contains cdna coding for poliovirus proteins 3d, 3c, 3b, and a portion of 3a protein sequence. immunofluorescence microscopy revealed that the majority of 3d (polymerase) was in the cytoplasm of recombinant ... | 1991 | 1654015 |
| functional expression of the human growth factor activatable na+/h+ antiporter (nhe-1) in baculovirus-infected cells. | we constructed a recombinant baculovirus, based on autographa californica nuclear polyhedrosis virus, containing the human na+/h+ antiporter cdna under control of the polyhedrin promoter. when infected with this recombinant baculovirus, the sf9 cell line, derived from spodoptera frugiperda, expresses a fully functional na+/h+ antiporter as measured by the generation of an amiloride-sensitive na+ influx in response to an acid load. the na+/h(+)-exchange activity, not detectable in sf9 cells, emer ... | 1991 | 1654101 |
| overexpression and characterization of the human mineralocorticoid receptor. | the full-length human renal mineralocorticoid receptor (hmr) has been overproduced in spodoptera frugiperda (sf9) insect cells using baculovirus-mediated expression. the overproduced hmr binds aldosterone with high affinity (kd = 1.36 nm) and has high affinity for cortisol, cortexolone, and progesterone. immunoprecipitation and immunoblot analysis of the recombinant hmr with mr-specific antibodies reveal three major protein bands with molecular masses of 115, 119, and 125 kda. hmr isoforms show ... | 1991 | 1655735 |