| expression and enzyme activity determination of human cyclooxygenase-1 and -2 in a baculovirus-insect cell system. | to develop an in vitro intact cell-based assay for screening selective cyclooxygenase inhibitors. | 2004 | 15301731 |
| identification and molecular cloning of insecticidal toxins from the venom of the brown spider loxosceles intermedia. | the venom of loxosceles intermedia was investigated for the presence of insecticidal toxins active against spodoptera frugiperda (lepdoptera: noctuidade), an insect that has caused great reductions in corn production in brazil. a combination of gel filtration (sephadex g-100) and ion-exchange chromatography (carboxymethyl cellulose, cm 52) resulted in four major fractions that were submitted to biological assay. fraction 4 was further purified by a reverse phase hplc (c18 column) resulting in pe ... | 2004 | 15302533 |
| recombinant destabilase-lysozyme: synthesis de novo in e. coli and action mechanism of the enzyme expressed in spodoptera frugiperda. | destabilase-lysozyme (dl) from salivary gland secretion of the medicinal leech (hirudo medicinalis) is as a member of the invertebrate lysozyme family, which sharply differs from other lysozyme families. in this study, dl lysozyme function was confirmed during expression of a gene encoding dl in escherichia coli. several constructs of the expression vectors pkk ompa and pet-3a with or without bacterial, leech, or yeast signal peptides (sp) were used. the use of a construct without signal peptide ... | 2004 | 15310278 |
| ubiquitin protein ligase activity of the anti-apoptotic baculovirus protein op-iap3. | the baculovirus inhibitor of apoptosis protein (iap) op-iap3 is required to prevent apoptosis during infection of insect cells by orgyia pseudotsugata m nucleopolyhedrovirus (opmnpv) and inhibits apoptosis when overexpressed in insect and mammalian cells. although previous reports have demonstrated that the ring domain is important for the anti-apoptotic function of op-iap3, the function of this domain in op-iap3 has not been studied. here, the ability of op-iap3 to function as an e3 ubiquitin p ... | 2004 | 15325084 |
| a uniform procedure for the purification of cdk7/cych/mat1, cdk8/cycc and cdk9/cyct1. | we have established a uniform procedure for the expression and purification of the cyclin-dependent kinases cdk7/cych/mat1, cdk8/cycc and cdk9/cyct1. we attach a his(6)-tag to one of the subunits of each complex and then co-express it together with the other subunits in spodoptera frugiperda insect cells. the cdk complexes are subsequently purified by ni(2+)-nta and mono s chromatography. this approach generates large amounts of active recombinant kinases that are devoid of contaminating kinase ... | 2004 | 15328539 |
| trypsin inhibitor from poecilanthe parviflora seeds: purification, characterization, and activity against pest proteases. | plants synthesize a variety of molecules, including proteinaceous proteinase inhibitors, to defend themselves of being attacked by insects. in this work, a novel trypsin inhibitor (ppti) was purified from the seeds of the native brazilian tree poecilanthe parviflora (benth) (papilioinodeae, leguminosae) by gel filtration chromatography on a sephadex g-100 followed by superdex g75 chromatography (fplc), sepharose 4b-trypsin column, and fractionated by reversed-phase hplc on a c-18 column. sds-pag ... | 2004 | 15328890 |
| genetic structure of a spodoptera frugiperda nucleopolyhedrovirus population: high prevalence of deletion genotypes. | a nicaraguan field isolate (sfnic) of spodoptera frugiperda nucleopolyhedrovirus was purified by plaque assay on sf9 cells. nine distinct genotypes, a to i, were identified by their restriction endonuclease profiles. variant sfnic-b was selected as the standard because its restriction profile corresponded to that of the wild-type isolate. physical maps were generated for each of the variants. the differences between variants and the sfnic-b standard were confined to the region between map units ... | 2004 | 15345446 |
| the nramp orthologue of cryptococcus neoformans is a ph-dependent transporter of manganese, iron, cobalt and nickel. | cryptococcus neoformans is an important human opportunistic pathogen and a facultative intracellular parasite, particularly in hiv-infected individuals. little is known about metal ion transport in this organism. c. neoformans encodes a single member of the nramp (natural resistance-associated macrophage protein) family of bivalent cation transporters, known as cramp, which we have cloned and expressed in xenopus laevis oocytes and spodoptera frugiperda sf 21 insect cells. cramp induces saturabl ... | 2005 | 15350193 |
| improved pc1/3 production through recombinant expression in insect cells and larvae. | protein convertase 1/3 is a serine endoproteinase present in the regulated secretory pathway of endocrine and neuroendocrine cells. it is responsible for the processing of numerous prohormones and proneuropeptides into their biologically active moieties, often following cleavage at pairs of basic residues. the determination of its three-dimensional structure, as well as the understanding of its enzymatic properties, would greatly benefit from the production and availability of large amounts of r ... | 2004 | 15358360 |
| characterization and localization of the unique marek's disease virus type 2 orf873 gene product. | studies on marek's disease virus (mdv)-unique genes are important for understanding the biological nature of the virus. based on complete dna sequence analyses of the mdv genomes, the mdv genomes contain presumably at least five mdv-unique genes, which are commonly conserved among the three mdv serotypes. a recombinant baculovirus that contains the mdv serotype 2 (mdv2)-unique gene, orf873, under the polyhedrin promoter was constructed and designated racorf873. polyclonal and monoclonal antibodi ... | 2004 | 15365234 |
| discovery of brl 50481 [3-(n,n-dimethylsulfonamido)-4-methyl-nitrobenzene], a selective inhibitor of phosphodiesterase 7: in vitro studies in human monocytes, lung macrophages, and cd8+ t-lymphocytes. | the biochemical and pharmacological characteristics in human proinflammatory cells of brl 50481 [3-(n,n-dimethylsulfonamido)-4-methyl-nitrobenzene], a novel and selective inhibitor of phosphodiesterase (pde) 7, are described. brl 50481 inhibited the activity of hrpde7a1 expressed in baculovirus-infected spodoptera frugiperda 9 cells in a competitive manner (ki value of 180 nm) and was 416 and 1884 times less potent against pde3 and 38 and 238 times less potent against pde4 at a substrate concent ... | 2004 | 15371556 |
| characterization of a novel peptide with allatotropic activity in the fall armyworm spodoptera frugiperda. | a cdna that encodes 53 amino acids, including one copy of the rvrgnpiscf-oh peptide, was cloned from spodoptera frugiperda. this peptide strongly stimulates the synthesis and release of juvenile hormone (jh) in vitro by the corpora allata (ca) of s. frugiperda and was code-named spofr-at 2. northern blotting and reverse transcriptase polymerase chain reaction (rt-pcr) analyses revealed that the preprohormone is expressed as one transcript in the brain, midgut (mg) and ovary (ov) in a tissue- and ... | 2004 | 15380923 |
| formulation with an optical brightener does not increase probability of developing resistance to spodoptera frugiperda nucleopolyhedrovirus in the laboratory. | stilbene-derived optical brighteners can markedly enhance the insecticidal activity of certain baculoviruses. we evaluated the influence of an optical brightener on the rate at which spodoptera frugiperda (j.e. smith) (lepidoptera: noctuidae) developed resistance to nucleopolyhedrovirus (sfmnpv). two laboratory colonies of s. frugiperda were inoculated with an lc50 of sfmnpv, in the absence or presence of the optical brightener tinopal lpw (0.1%), over a period of two and 11 generations, in the ... | 2004 | 15384328 |
| virus entry or the primary infection cycle are not the principal determinants of host specificity of spodoptera spp. nucleopolyhedroviruses. | the multicapsid nucleopolyhedroviruses (npvs) of spodoptera exigua (semnpv), spodoptera frugiperda (sfmnpv), and spodoptera littoralis (splinpv) are genetically similar (78 % similarity) but differ in their degree of host specificity. infection by each of the three npvs in these three spodoptera host species was determined by oral inoculation of larvae with occlusion bodies (obs) or intrahaemocoelic injection with occlusion derived virions (odvs). rt-pcr analysis of total rna from inoculated ins ... | 2004 | 15448346 |
| the role of carboxyl, guanidine and imidazole groups in catalysis by a midgut trehalase purified from an insect larvae. | a trehalase (ec 3.2.1.28) of 67 kda was purified to homogeneity from the midgut of spodoptera frugiperda (lepidoptera) larvae. the enzyme is inhibited by toxic beta-glucosides produced by plants (amygdalin, prunasin, salicin and phlorezin) and by their aglycones (mandelonitrile, phloretin). from kcat and km values determined in different phs, the pka values of catalytic essential groups were calculated (pka = 4.5 and pka = 8.0). these pka values agree with the ones determined from enzyme chemica ... | 2004 | 15475303 |
| cytotoxicity induced by nivalenol, deoxynivalenol, and fumonisin b1 in the sf-9 insect cell line. | the toxicity of the mycotoxins nivalenol (niv), deoxynivalenol (don), and fumonisin b1 (fb1) was studied in the lepidopteran spodoptera frugiperda (sf-9) cells, by the trypan blue dye-exclusion and 3-(4,5-dimethylthiozole-2-yl)-2,5-biphenyl tetrazolium bromide (mtt) tests, uptake analyses of cytotoxicity, and cell metabolism, respectively. deoxyribonucleic acid analysis by flow cytometry was used to identify apoptosis and cell cycle distribution. after 48 h of exposure, the mtt and trypan blue d ... | 2004 | 15479121 |
| comparative physical localization of maize mir1 gene in zea mays l. and coix lacryma-jobi l. | the maize gene mir1 encoded a cystein proteinase which is resistant to fall armyworm. previously, rflp map indicated that the mir1 was mapped on chromosome 6. however, physical location of the mir1 gene on chromosome 6 has not been reported. in this study, the mir1 gene was physically located on the short arm of metaphase and pachytene chromosome 6 by dual-color fluorescence in situ hybridization (fish) with 45s rdna as a reference marker. the results of southern blotting suggested that there we ... | 2004 | 15487499 |
| the murine thymic stromal lymphopoietin receptor is partially expressed with an extracellular c-terminus. | the gene encoding the murine thymic stromal lymphopoietin receptor was expressed in lepidopteran insect cells using the baculovirus expression vector system. the corresponding gene was inserted under the polyhedrin promoter of the autographa californica nuclear polyhedrosis virus and expressed with an n-terminal poly-histidine tag and a c-terminal flag-tag in the spodoptera frugiperda insect cell line sf9 during viral infection. flow cytometer analysis of cells infected with the produced recombi ... | 2005 | 15488943 |
| investigation of the substrate specificity of a beta-glycosidase from spodoptera frugiperda using site-directed mutagenesis and bioenergetics analysis. | the specificity of the spodoptera frugiperda digestive beta-glycosidase (sfbetagly50) for fucosides, glucosides and galactosides is determined by noncovalent interactions of glycone 6-oh and glycone 4-oh with the active-site residues q39 and e451. site-directed mutagenesis and enzyme steady-state kinetics were described, showing that replacement of e451 with glutamine increased the preference of sfbetagly50 for glucosides in comparison to galactosides, whereas replacing e451 with serine had the ... | 2004 | 15511222 |
| baculovirus proteins ie-1, lef-3, and p143 interact with dna in vivo: a formaldehyde cross-linking study. | ie-1, lef-3, and p143 are three of six proteins encoded by autographa californica nucleopolyhedrovirus (acmnpv) essential for baculovirus dna replication in transient replication assays. ie-1 is the major baculovirus immediate early transcription regulator. lef-3 is a single-stranded dna binding protein (ssb) and p143 is a dna helicase protein. to investigate their interactions in vivo, we treated acmnpv-infected spodoptera frugiperda cells with formaldehyde and separated soluble proteins from c ... | 2004 | 15518813 |
| porcine purple acid phosphatase: heterologous expression, characterization, and proteolytic analysis. | uteroferrin is an iron-binding glycoprotein, which is abundantly synthesized in porcine uterine glandular endometrium and believed to be involved in maternal/fetal iron transport. in the present study, uteroferrin has been cloned and functionally expressed using baculovirus-infected insect host cells spodoptera frugiperda. the work also addresses the possible role of proteolytic cleavage to facilitate the release of uteroferrin-bound iron. the enzyme secreted in culture medium exhibits a molecul ... | 2004 | 15519293 |
| molecular characterization of brevibacillus laterosporus and its potential use in biological control. | thirty-three strains of brevibacillus laterosporus, including three novel strains isolated from brazilian soil samples, were examined for genetic variability by the use of different pcr-based methods. molecular markers that could characterize bacterial strains with regards to their pathogenic potential were investigated. in addition, toxicity was assessed by the use of insects belonging to the orders lepidoptera and coleoptera and the mollusk biomphalaria glabrata. among the targets tested, biom ... | 2004 | 15528531 |
| effect of expression of manganese superoxide dismutase in baculovirus-infected insect cells. | it has previously been demonstrated that baculovirus infection of the spodoptera frugiperda sf-9 (sf-9) and trichoplusia ni bti-tn-5b1-4 (tn-5b1-4) insect cell lines leads to oxidative stress as measured by protein and membrane lipid oxidation and that this oxidative damage contributes to cell death. as a result of these findings, it was hypothesized that baculovirus infection stimulates superoxide radical (o(2)(.-)) synthesis in the mitochondria and that the resulting o(2)(.-) accumulation over ... | 2004 | 15531788 |
| functional analysis of human microsomal epoxide hydrolase genetic variants. | human microsomal epoxide hydrolase (ephx1) is active in the metabolism of many potentially carcinogenic or otherwise genotoxic epoxides, such as those derived from the oxidation of polyaromatic hydrocarbons. ephx1 is polymorphic and encodes allelic variation at least two amino acid positions, y113h and h139r. in a number of recent molecular epidemiological investigations, ephx1 polymorphism has been suggested as a susceptibility factor for several human diseases. to better evaluate the functiona ... | 2004 | 15535985 |
| the ecdysteroid udp-glucosyltransferase gene promoter from autographa californica multicapsid nucleopolyhedrovirus. | the ecdysteroid udp-glucosyltransferase (egt) gene promoter fragments of different lengths were generated from the genomic dna of the autographa californica multicapsid nucleopolyhedrovirus (acmnpv) by pcr. after being purified and enzymatic digestion, they were cloned into the pgem-3z vector for construction of reporter plasmids pacegt542-luc, pacegt309-luc and pacegt159-luc with the luciferase gene driven by the acmnpv egt promoter. the results of transient expression in the spodoptera frugipe ... | 2004 | 15540610 |
| plant-incorporated bacillus thuringiensis resistance for control of fall armyworm and corn earworm (lepidoptera: noctuidae) in corn. | fall armyworm, spodoptera frugiperda (j.e. smith), and corn earworm, helicoverpa zea (boddie), perennially cause leaf and ear damage to corn, zea mays l., in the southeastern united states. transgenic bacillus thuringiensis (bt) hybrids with the bt11, mon810, or 176 events expressing the cry1ab insecticidal endotoxin from were evaluated for control fall armyworm and corn earworm at seven locations in georgia during 1999 and 2000. corn was planted at the recommended time for each location and 1 a ... | 2004 | 15568349 |
| relative concentration of cry1a in maize leaves and cotton bolls with diverse chlorophyll content and corresponding larval development of fall armyworm (lepidoptera: noctuidae) and southwestern corn borer (lepidoptera: crambidae) on maize whorl leaf profiles. | to manage insect resistance to transgenic crops that express insecticidal proteins from bacillus thuringiensis (bt) berliner, the u.s. environmental protection agency recommends a refuge-based insect resistance management strategy where a percentage of non-bt (refuge) crop is grown in proximity to a bt-expressing crop. an important requirement for this strategy is that the toxin exists at a high effective dose for control of the target pest(s), so that heterozygous individuals in the population ... | 2004 | 15568367 |
| characterization of the replication of a baculovirus mutant lacking the dna polymerase gene. | in a previous study, the dna polymerase gene (dnapol) of autographa californica multiple nucleopolyhedrovirus (acmnpv) was identified as one of six genes required for plasmid replication in a transient replication assay (m. kool, c. ahrens, r.w. goldbach, g.f. rohrmann, j.m. vlak, identification of genes involved in dna replication of the autographa californica, proc. natl. acad. sci. u.s.a. 91, (1994) 11212-11216); however, another study based on a similar approach reported that the virally enc ... | 2005 | 15582664 |
| expression of the retrovirus gypsy gag in spodoptera frugiperda cell culture with the recombinant baculovirus. | | 2004 | 15584516 |
| alzheimer amyloid precursor aspartyl proteinase activity in chapso homogenates of spodoptera frugiperda cells. | presenilins are polytopic, integral proteins that control intramembranous proteolysis at the "gamma-" and "epsilon-" cleavage sites of the alzheimer amyloid-beta precursor protein (app) to yield amyloid-beta peptide (abeta) and the app intracellular domain (aicd). we have overexpressed a constitutively active, pathogenic form of ps1 (known as ps1 delta exon 9) together with its substrate, app-c99, in spodoptera frugiperda (sf9) cells. sf9 cells have been reported to lack endogenous gamma-secreta ... | 2004 | 15592142 |
| expression and purification of active plant diphosphonucleotide phosphatase/phosphodiesterase from baculovirus-infected insect cells. | we have previously purified and characterized a diphosphonucleotide phosphatase/phosphodiesterase (ppd1) from yellow lupin seeds. this report describes an efficient strategy for overexpression in baculovirus-infected spodoptera frugiperda sf9 cells and purification of a functional ppd1 enzyme. we tested six variants of recombinant ppd1, differing in secretion peptides, fusion tags, and promoters. the highest expression level of the active ppd1 was achieved when the native signal peptide and the ... | 2005 | 15596367 |
| identification, cloning and expression of a cry1ab cadherin receptor from european corn borer, ostrinia nubilalis (hubner) (lepidoptera: crambidae). | transgenic corn expressing the cry1ab toxin from bacillus thuringiensis is highly toxic to european corn borer, ostrinia nubilalis, larvae. a putative cry1ab receptor (onbt-r(1)) molecule was cloned and sequenced from a cdna library prepared from midgut tissue of o. nubilalis larvae. the 5.6 kb gene is homologous with a number of cadherin genes identified as cry1 binding proteins in other lepidopterans. brush border membrane vesicles were prepared using dissected midguts from late instars. a 220 ... | 2005 | 15607653 |
| the structures of the unique sulfotransferase retinol dehydratase with product and inhibitors provide insight into enzyme mechanism and inhibition. | the structure of retinol dehydratase (dhr) from spodoptera frugiperda, a member of the sulfotransferase superfamily, in complexes with the inactive form of the cofactor pap 3'-phosphoadenosine 5'-phosphate (pap) and (1) the product of the reaction with retinol anhydroretinol (ar), (2) the retinoid inhibitor all-trans-4-oxoretinol (or), and (3) the potent steroid inhibitor androsterone (and) have been determined and compared to the enzyme complex with pap and retinol. the structures show that the ... | 2004 | 15608121 |
| nucleotide sequence and molecular characterization of the structural glycoprotein gp41 gene homologue of spodoptera litura nucleopolyhedrosis virus (spltnpv-i). | the structural glycoprotein gene gp41 homologue of spodoptera litura nucleopolyhedrosis virus (spltnpv-i *) was identified in the 4.0 kb ecori-l fragment of the viral genome. the nucleotide sequence of 2063 bp of this fragment revealed an open reading frame of 1014 nucleotides to encode a polypeptide of 337 amino acids. analysis of nucleotide and deduced amino acid sequences of the putative orf indicated its identity with gp41 protein of other baculoviruses sharing maximum homology with that of ... | 2004 | 15663006 |
| the deletion of the pif gene improves the biosafety of the baculovirus-based technologies. | our goal was to improve the biosafety of baculovirus-based technologies by deleting the pif (per os infectivity factor) gene from baculovirus expression vectors. such a deletion would block transmission in nature without disturbing protein production. a pif deletion mutant of autographa californica multiplecapsid nucleopolyhedrovirus (acmnpv) was constructed and its infectivity to two host species was tested by oral or intrahemocoelic inoculation. virus replication after oral inoculation was mon ... | 2004 | 15664077 |
| species and cultivar influences on survival and parasitism of fall armyworm. | interactions between host plant resistance and biological control may benefit or hinder pest management efforts. turfgrass cultivars have rarely been tested for extrinsic resistance characteristics such as occurrence and performance of beneficial arthropods on plant genotypes with resistance to known turf pests. parasitism of fall armyworm, spodoptera frugiperda (j.e. smith), among six turfgrass genotypes was evaluated. the six grasses tested [sea isle-1 and 561-79 seashore paspalum, paspalum va ... | 2004 | 15666756 |
| developmental expression of a tyramine receptor gene in the brain of the honey bee, apis mellifera. | this study reveals that the tyramine receptor gene, amtyr1, is expressed in the developing brain, as well as in the brain of the adult worker honey bee. changes in levels of amtyr1 expression were examined using northern analysis. age-related increases in amtyr1 transcript levels were observed not only during metamorphic adult development, but also in the brain of the adult worker bee. rna in situ hybridization revealed the pattern of amtyr1 expression. cell bodies staining intensely for tyramin ... | 2005 | 15672398 |
| nucleotide sequence and transcriptional analysis of the pif gene of spodoptera frugiperda nucleopolyhedrovirus (sfmnpv). | defective viruses, not transmissible alone, increase the transmissibility of complete genotypes in natural populations of spodoptera frugiperda multicapsid nucleopolyhedrovirus (sfmnpv). the defective phenotype is associated with a 15 kb deletion, which includes the pif (per os infectivity factor) gene. the sequence of a 2.4 kb fragment that includes pif was determined. multiple transcripts encompassing pif were detected by northern blot analysis. rt-pcr and nuclease protection analysis demonstr ... | 2005 | 15681073 |
| receptor-mediated endocytosis and intracellular trafficking of lipoproteins and transferrin in insect cells. | while the intracellular pathways of ligands after receptor-mediated endocytosis have been studied extensively in mammalian cells, in insect cells these pathways are largely unknown. we transfected drosophila schneider line 2 (s2) cells with the human low-density lipoprotein (ldl) receptor (ldlr) and transferrin (tf) receptor (tfr), and used endocytosis of ldl and tf as markers. after endocytosis in mammalian cells, ldl is degraded in lysosomes, whereas tf is recycled. fluorescence microscopy ana ... | 2005 | 15681222 |
| the function of envelope protein p74 from autographa californica multiple nucleopolyhedrovirus in primary infection to host. | this research investigated the function of envelope protein p74 of autographa californica multiple nuclear polyhedrosis virus (acmnpv) in primary infection to host. a p74-inactivation recombinant baculovirus, rac-gfp(delta) p74, was constructed by inserting gfp driven by acmnpv polyhedrin promoter into the p74 locus of acmnpv genome. bioassays showed that the p74-null occlusion bodies (obs) failed to infect its natural host larvae, spodoptera exigua, per os, while the p74-null budded virus (bvs) ... | 2005 | 15744572 |
| optical brighteners do not influence covert baculovirus infection of spodoptera frugiperda. | covert infection with spodoptera frugiperda multiple nucleopolyhedrovirus, detected by reverse transcription-pcr of virus gene transcripts (ie-0 and polh), was not significantly affected by the presence of an optical brightener (tinopal unpa-gx), indicating no change in virus virulence. detection of the covert infection was dependent on insect life stage and the viral mrna used for diagnosis. | 2005 | 15746377 |
| cry1ab protoxin effects on spodoptera frugiperda (lepidoptera: noctuidae) and its parasitoid cotesia marginiventris (hymenoptera: braconidae). | | 2004 | 15759410 |
| cloning and characterization of the g protein betagamma subunits from trichoplusia ni (high five cells). | baculoviral-mediated expression in insect cells has become a method of choice where high-level protein expression is desired and where expression in escherichia coliform (e. coli.) is unsuitable. genes of interest are inserted into the baculoviral genome of autographa californica nuclear polyhedrosis virus (acnpv) under the extremely strong, but very late polyhedron gene (polh). the preferred host lines are derived from spodoptera frugiperda (sf9 or sf21) or tricoplusia ni (high five, invitrogen ... | 2005 | 15763469 |
| spodoptera frugiperda pheromone lures to avoid nontarget captures of leucania phragmatidicola. | we confirmed that commercial three- or four-component spodoptera frugiperda (j.e. smith) pheromone lures had a high nontarget capture rate for leucania phragmatidicola guenée, which compromised monitoring efforts in the northeastern united states. we compiled taxonomic features to distinguish l. phragmatidicola from s. frugiperda, and we compared five new lures. s. frugiperda catch specificity was improved by removing (z)-11-hexadecen-1-ol acetate (z11-16:ac), which attracted l. phragmatidicola. ... | 2005 | 15765666 |
| biological and molecular characterization of a multicapsid nucleopolyhedrovirus from thysanoplusia orichalcea (l.) (lepidoptera: noctuidae). | a multicapsid nucleopolyhedrovirus (thormnpv) that was co-isolated with a single nucleocapid thorsnpv from mixed infected larvae of thysanoplusia orichalcea l. (lepidoptea: noctuidae) is characterized. scanning electron microscopy of thormnpv showed a dodecahedral-shaped occlusion body (ob). the occluded virions contained one to as many as eight nucleocapsids/virion. virion band profiles in gradient centrifugation were consistent in at least 10 rounds of centrifugation from different virion samp ... | 2005 | 15766929 |
| recombinant rotat 1.2 variable surface glycoprotein as antigen for diagnosis of trypanosoma evansi in dromedary camels. | the transcript encoding a predominant trypanosoma evansi variable surface glycoprotein rotat 1.2 was cloned and expressed as a recombinant protein in spodoptera frugiperda and trichoplusia ni (insect) cells. its potential as an antigen for specific detection of antibody in serum of dromedary camels affected by surra, was evaluated. in elisa, the reactivity of the recombinant rotat 1.2 vsg was similar to that of native rotat 1.2 vsg. an indirect agglutination reagent was therefore prepared by cou ... | 2005 | 15777921 |
| expression of a toxoneuron nigriceps polydnavirus-encoded protein causes apoptosis-like programmed cell death in lepidopteran insect cells. | the polydnavirus toxoneuron nigriceps bracovirus (tnbv) is an obligate symbiont associated with the braconid wasp t. nigriceps, a parasitoid of heliothis virescens larvae. previously, to identify polydnavirus genes that allow parasitization by altering the host immune and endocrine systems, expression patterns of tnbv genes from parasitized h. virescens larvae were analysed and cdnas were obtained. to study the function of the protein from one such cdna, tnbv1, overexpression of the protein was ... | 2005 | 15784889 |
| antiinsectan decaturin and oxalicine analogues from penicillium thiersii. | three new oxalicine and decaturin analogues (6-8) have been isolated from organic extracts of penicillium thiersii, along with the known compounds oxalicines a and b. these compounds (4-8) are members of a group of unique natural products containing terpenoid and pyridine moieties. the structures of the new compounds were elucidated by analysis of nmr and ms data. most of these metabolites exhibit potent antiinsectan activity against the fall armyworm (spodoptera frugiperda). | 2005 | 15787428 |
| the autographa californica multiple nucleopolyhedrovirus ie0-ie1 gene complex is essential for wild-type virus replication, but either ie0 or ie1 can support virus growth. | the immediate-early ie0-ie1 gene complex expresses the only baculovirus spliced gene that produces an alternate protein product. autographa californica multiple nucleopolyhedrovirus (acmnpv) ie1 is a potent transcriptional transactivator that is essential for viral replication in transient assays. ie1 contains 582 amino acids that are arranged into different domains, including an acidic activation domain at the n terminus, a dna binding domain, and an oligomerization domain at the c terminus. ie ... | 2005 | 15795248 |
| antimicrobial activity of conditioned medium fractions from spodoptera frugiperda sf9 and trichoplusia ni hi5 insect cells. | concentrated conditioned medium (cm) fractions from spodoptera frugiperda sf9 and trichoplusia ni cells, eluting from a gel filtration column at around 10 kda, were found to exhibit strong antibacterial activity against bacillus megaterium and escherichia coli. the b. megaterium cells incubated in the cm fraction from sf9 cells rapidly lost viability: after 8 min the viability had decreased to 0.7%, as compared with the control. addition of the cm fraction to e. coli cells resulted in a less dra ... | 2005 | 15806353 |
| systematic mutagenesis of potential glycosylation sites of lysosomal acid lipase. | lysosomal acid lipase (lal; ec 3.1.1.13) is a key enzyme in the intracellular lipid metabolism. it hydrolyzes exogenous triglycerides and cholesterol esters taken up by various cell types. lal has six potential n-glycosylation sites and one potential o-glycosylation site. elimination of each of the six asn-(x)-ser/thr sites by site-directed mutagenesis and expression in baculovirus-infected spodoptera frugiperda cells resulted in two single-mutant enzymes without lipolytic activities (n134q and ... | 2005 | 15809341 |
| mammalian-like nonsialyl complex-type n-glycosylation of equine gonadotropins in mimic insect cells. | recombinant equine luteinizing hormone/chorionic gonadotropin (elh/cg) was expressed in mimic insect cells, that are commercial stably transformed spodoptera frugiperda (sf9) cells expressing five mammalian genes encoding glycosyltransferases involved in the synthesis of complex-type monosialylated n-glycans. we previously showed that it exhibited no in vivo bioactivity although expressing full in vitro bioactivity, and it was suspected that this was because of insufficient sialylation of elh/cg ... | 2005 | 15814822 |
| novel macula-like virus identified in bombyx mori cultured cells. | we identified a novel, 6,513-bp-long rna, termed bombyx mori macula-like latent virus (bmmlv) rna, which abundantly expressed in b. mori cultured bmn cells. bmmlv rna potentially encodes two proteins, putative rna replicase and coat protein, which share structural features and sequence similarities with those of a plant rna virus, the genus maculavirus. northern blot analysis showed that two transcripts were expressed in bmn cells: a 6.5-kb-long rna, which contains both putative rna replicase an ... | 2005 | 15827172 |
| physical and partial genetic map of spodoptera frugiperda nucleopolyhedrovirus (sfmnpv) genome. | a nicaraguan isolate of spodoptera frugiperda multicapsid nucleopolyhedrovirus (sfmnpv) is undergoing field trials for control of this pest in the americas. this isolate is composed of multiple genotypes, some of which are deletion mutants. identification of the genetic changes in deleted genotypes cannot be accomplished without the construction of a detailed physical map. in the present study, combinations of restriction endonuclease analysis and southern blot analysis was performed. this map w ... | 2005 | 15830159 |
| spodoptera frugiperda resistance to oral infection by autographa californica multiple nucleopolyhedrovirus linked to aberrant occlusion-derived virus binding in the midgut. | spodoptera frugiperda larvae are highly resistant to oral infection by autographa californica multiple nucleopolyhedrovirus (acmnpv) (ld(50), approximately 9200 occlusions), but extremely susceptible to budded virus within the haemocoel (ld(50), <1 p.f.u.). the inability of acmnpv occlusion-derived virus (odv) to establish primary infections readily within midgut cells accounts for a major proportion of oral resistance. to determine whether inappropriate binding of acmnpv odv to s. frugiperda mi ... | 2005 | 15831946 |
| insecticidal activity of the pyrimidine nucleoside analogue (e)-5-(2-bromovinyl)-2'-deoxyuridine (bvdu). | the insecticidal activity of the antiherpetic agent (e)-5-(2-bromovinyl)-2'-deoxyuridine (bvdu) was assessed in in vivo assays against the fall armyworm, spodoptera frugiperda (je smith) (lepidoptera, noctuidae). bvdu, mixed into an artificial diet, caused a variety of effects, depending on the concentration used. compared with controls, food intake was lower, larval growth was retarded and larval development was prolonged. the treated larvae formed smaller pupae and the hatching moths often sho ... | 2005 | 15838935 |
| manoyl oxide diterpenoids from grindelia scorzonerifolia. | two new norditerpenoids, 4beta-hydroxy-19-normanoyl oxide (1) and 4alpha-hydroxy-18-normanoyl oxide (2), the new 18-o-alpha-l-arabinopyranosylmanoyl oxide (3a), and the known diterpenoids jhanol (4) and 18-hydroxy-13-epi-manoyl oxide (5) were isolated, together with other common plant constituents from an argentine collection of grindelia scorzonerifolia. the structures of the new compounds were established by extensive 1d and 2d nmr techniques and chemical transformations. structural features o ... | 2005 | 15844947 |
| expression of a novel recombinant stem cell factor/macrophage colony-stimulating factor fusion protein in baculovirus-infected insect cells. | a novel human stem cell factor (scf)/macrophage colony-stimulating factor (m-csf) fusion protein gene was constructed, in which the coding regions of human scf cdna (1-165aa) and the truncated m-csf cdna (1-149aa) were connected by a linker sequence encoding a short peptide ggggsggggsgg. the scf/m-csf gene was cloned into baculovirus transfer vector pvl1392 under the control of polyhedrin promoter and expressed in the sf9 cells (spodoptera frugiperda). sds-page and western blot analysis showed t ... | 2005 | 15866728 |
| insights into the central metabolism of spodoptera frugiperda (sf-9) and trichoplusia ni bti-tn-5b1-4 (tn-5) insect cells by radiolabeling studies. | the insect cell baculovirus expression vector system (bevs) is one of the most commonly used expression systems for recombinant protein production. this system is also widely used for the production of recombinant virus and virus-like particles. although several published reports exist on recombinant protein expression using insect cells, information dealing with their metabolism in vitro is relatively scarce. in this work we have analyzed the metabolism of glucose and glutamine, the main carbon ... | 2005 | 15903243 |
| purification and characterization of an anti-apoptotic protein isolated from lonomia obliqua hemolymph. | previously it was reported that supplementation of insect cell culture with lonomia obliqua hemolymph could extend culture longevity (maranga et al. biotechnol. prog. 2003, 19, 58-63). in this work the anti-apoptotic properties of this hemolymph in spodoptera frugiperda (sf-9) cell culture were investigated. the presence or absence of apoptotic cells was characterized by light microscopy, flow cytometry, and agarose gel electrophoresis. hemolymph was fractionated by several ion exchange and gel ... | 2005 | 15903246 |
| production of recombinant adeno-associated viral vectors using a baculovirus/insect cell suspension culture system: from shake flasks to a 20-l bioreactor. | production of recombinant adeno-associated viral vectors using a baculovirus/insect cell system at various scales is presented. shake flask studies were conducted to assess conditions to be used in bioreactors. two insect cell lines, trichoplusia ni (h5) and spodoptera frugiperda (sf9), were compared for their ability to produce raav-2 after infection with recombinant baculoviruses coding for the essential components of the vector. the effect of varying the ratio between individual baculoviruses ... | 2005 | 15903253 |
| cyp2k6 from zebrafish (danio rerio): cloning, mapping, developmental/tissue expression, and aflatoxin b1 activation by baculovirus expressed enzyme. | a full-length zebrafish (danio rerio) cytochrome p450 (cyp) 2k6 cdna, was obtained (genbank accession no. af283813) through polymerase chain reaction cloning using degenerated primers based on a consensus cyp2 sequence and the heme-binding domain. this first cyp2k family member cloned from zebrafish had 1861 bp which contained 27 bp of 5'-untranslated region (5'-utr), an open reading frame (orf) of 1518 bp, and a 300 bp 3'-utr with a poly a tail. the deduced 506 amino acid sequence of cyp2k6 had ... | 2005 | 15907766 |
| in vivo bioinsecticidal activity toward ceratitis capitata (fruit fly) and callosobruchus maculatus (cowpea weevil) and in vitro bioinsecticidal activity toward different orders of insect pests of a trypsin inhibitor purified from tamarind tree (tamarindus indica) seeds. | a proteinaceous inhibitor with high activity against trypsin-like serine proteinases was purified from seeds of the tamarind tree (tamarindus indica) by gel filtration on shephacryl s-200 followed by a reverse-phase hplc vidac c18 tp. the inhibitor, called the tamarind trypsin inhibitor (tti), showed a mr of 21.42 kda by mass spectrometry analysis. tti was a noncompetitive inhibitor with a ki value of 1.7 x 10(-9) m. in vitro bioinsecticidal activity against insect digestive enzymes from differe ... | 2005 | 15913299 |
| baculovirus-based expression of an insect viral protein in 12 different insect cell lines. | the ability of 12 unique lepidopteran insect cell lines from anticarsia gemmatalis, heliothis virescens, lymantria dispar (two lines), mamestra brassica, plutella xylostella, spodoptera frugiperda (two lines), and trichoplusia ni (three lines) to support production of a recombinant polydnavirus (pdv) protein (gipdv 1.1) expressed using the bac-to-bac baculovirus expression system was examined. polydnavirus gene gipdv 1.1 was cloned into the pfastbac baculovirus vector under the control of the po ... | 2005 | 15926859 |
| a viral caspase contributes to modified apoptosis for virus transmission. | the spodoptera frugiperda ascovirus, a dna virus that attacks lepidopterans, codes for an executioner caspase synthesized by 9 h after infection of sf21 cells. this caspase alone induces apoptosis in insect cells and, during viral replication in vivo, contributes to a novel cell cleavage process in which developing apoptotic bodies are rescued by the virus and differentiate to form large vesicles in which virions assemble. these viral vesicles disseminate to the blood, where they are acquired du ... | 2005 | 15933068 |
| immunogenicity of baculovirus expressed recombinant proteins of japanese encephalitis virus in mice. | genes encoding for the premembrane and envelope (prme), envelope (e) and nonstructural protein (ns1) of japanese encephalitis virus (jev) were cloned. each protein was expressed in baculovirus expression system. of the three proteins expressed in baculovirus system, only prme had hemagglutination activity. the prme (72 and 54 kda), e (54 kda) and ns1 (46 kda) proteins could be detected by western blotting in the recombinant virus infected cells. immunogenicity of the recombinant proteins obtaine ... | 2005 | 15933432 |
| development of a novel serum-free freezing medium for mammalian cells using the silk protein sericin. | cryopreservation is a pivotal process in cellular engineering for creating a continuous source of generated functional cell lines and for the convenience of various medical treatments that involve cell culture. fbs (fetal bovine serum) supplemented with 10% (v/v) dmso is extensively used as a freezing medium for mammalian cells using conventional methods. however, fbs should ideally be avoided, owing to serious concerns regarding bovine spongiform encephalopathy and other infections such as viru ... | 2005 | 15943583 |
| grayanane diterpenoids from the flowers of rhododendron molle with cytotoxic activity against a spodoptera frugiperda cell line. | two new grayanane diterpenoids, rhodomolins a (1) and b (2), together with two known diterpenoids, rhodomolleins i and rhodojaponin iii, were isolated from the flowers of rhododendron molle. the structures of 1 and 2 were elucidated on the basis of interpretation of spectroscopic data. all compounds were evaluated for cytotoxic activity against the spodoptera frugiperda cell line sf-9 and gave ic(50) values in the range 12-80 microg/ml. | 2005 | 15974620 |
| purification and partial characterization of r5, r5x4, and x4 hiv-1 subtype c envelope glycoproteins expressed in insect cells. | biological properties of four recombinant hiv-1 subtype c envelope glycoproteins from viruses with different phenotypic characteristics (ccr5 and/or cxcr4-utilizing) were investigated. the gp160 genes were cloned, expressed in spodoptera frugiperda insect cells, purified, and their biological characteristics were examined. the conformational and functional integrity of the hiv-1 subtype c rgp160 was intact since they reacted with the a32, c11, igg1b12, 7b2, and 17b conformational dependant monoc ... | 2005 | 15977243 |
| cell-cycle-dependent resistance to bacillus thuringiensis cry1c toxin in sf9 cells. | the sf9 cell line, derived from the moth spodoptera frugiperda, is highly and specifically sensitive to the bacillus thuringiensis cry1c toxin. upon exposure to cry1c, ionic pores are formed in the plasma membrane leading to cell swelling and death. here, we describe a unique transient tolerance to cry1c of dividing cells, which allowed completion of the division process in the presence of cry1c. correlatively, arresting the cells at g2-m phase by nocodazole treatment rendered them insensitive t ... | 2005 | 15985466 |
| protein response of insect cells to bioreactor environmental stresses. | protein expression of spodoptera frugiperda (sf9) insect cells was characterized upon exposure to environmental stresses typically present in bioreactors including heat shock, oxygen deprivation, shear stress, change of ph, and salinity or ethanol shock. this study fills the void in knowledge as to how bioreactor hydrodynamics, anoxia, small changes in ph as well as salinity alterations due to ph control or exposure to ethanol used in asepsis treatments affect protein expression in sf9 cells. he ... | 2005 | 16002170 |
| the immediate-early protein ie0 of the autographa californica nucleopolyhedrovirus is not essential for viral replication. | the role of the autographa californica multiple nucleopolyhedrovirus (acmnpv) immediate-early protein ie0 in the baculoviral infection is not clear. in this study, we constructed the recombinant virus vacdeltaie0 null for ie0 expression by targeted mutagenesis replacing exon0 with the cat gene. we found that vacdeltaie0 replicated efficiently in spodoptera littoralis sl2 cells, which are poorly permissive for acmnpv. in contrast, in spodoptera frugiperda sf9 cells, which are fully permissive for ... | 2005 | 16014970 |
| enhancement of cauliflower mosaic virus 35s promoter in insect cells infected with baculovirus. | we happened to discover that the cauliflower mosaic virus (camv) 35s promoter inserted into a recombinant autographa californica multicapsid nucleopolyhedrovirus (racmnpv) was strongly activated during the replication of the recombinant virus in spodoptera frugiperda (sf9) cells. the expression of the luciferase gene from the 35s promoter in racmnpv was remarkably increased late in infection and was resistant to alpha-amanitin treatment. primer extension indicated that transcriptional initiation ... | 2005 | 16022899 |
| proteomic analysis of trpc5- and trpc6-binding partners reveals interaction with the plasmalemmal na(+)/k(+)-atpase. | mammalian transient receptor potential canonical (trpc) genes encode a family of nonselective cation channels that are activated following stimulation of g-protein-coupled membrane receptors linked to phospholipase c. in drosophila photoreceptor cells, trp channels are found in large, multimolecular signaling complexes in association with the pdz-containing scaffolding protein, inad. a similar mammalian trpc "signalplex" has been proposed, but has yet to be defined. in the present study, affinit ... | 2005 | 16025302 |
| behavior of trichogramma atopovirilia oatman & platner and t. pretiosum riley (hymenoptera: trichogrammatidae) on spodoptera frugiperda (j. e. smith) (lepidoptera: noctuidae) egg masses. | in this study we analyzed the impact of physical barriers of spodoptera frugiperda (j. e. smith) egg-masses on the behavior of trichogramma atopovirilia oatman & platner and trichogramma pretiosum riley. the duration of drumming, drilling, oviposition, period spent over the egg-mass, and interval between parasitized eggs were timed, and the number of parasitized eggs were recorded. the presence of scales on the egg-masses caused a significant increase in the time spent by both parasitoids on eac ... | 2005 | 16025898 |
| genetically engineered horseradish peroxidase for facilitated purification from baculovirus cultures by cation-exchange chromatography. | an engineered horseradish peroxidase isozyme c (hrp c) gene was constructed by the addition of a 6xarg fusion tail to 6xhis-hrp c by the pcr strategy. the 6xhis-6xarg-hrp c cdna was expressed in the sf9 insect cell line from spodoptera frugiperda infected with autographa californica nuclear polyhedrosis virus. the recombinant peroxidase isoelectric point was 9.5 as judged by isoelectric focusing and was purified directly from the culture medium at day-6 post-infection by cation-exchange chromato ... | 2005 | 16038999 |
| high yield of biologically active recombinant human amelogenin using the baculovirus expression system. | the amelogenins are secreted by the ameloblast cells of developing teeth; they constitute about 90% of the enamel matrix proteins and play an important role in enamel biomineralization. recent evidence suggests that amelogenin may also be involved in the regeneration of the periodontal tissues and that different isoforms may have cell-signalling effects. during enamel development and mineralization, the amelogenins are lost from the tissue due to sequential degradation by specific proteases, mak ... | 2006 | 16055347 |
| development of a bi-cistronic baculovirus expression vector by the rhopalosiphum padi virus 5' internal ribosome entry site. | a bi-cistronic baculovirus transfer vector was constructed based on the 5'utr internal ribosome entry site (ires) of the rhopalosiphum padi virus (rhpv). recombinant baculoviruses containing the red fluorescent protein gene and green fluorescent protein gene flanking the rhpv 5'utr ires can simultaneously produce dual fluorescence in recombinant virus-infected spodoptera frugiperda 21 cells (sf21) under the control of a polyhedrin promoter. quantization by fluorescence spectrophotometry of the f ... | 2005 | 16084836 |
| functional importance of deletion mutant genotypes in an insect nucleopolyhedrovirus population. | a nicaraguan isolate of a nucleopolyhedrovirus (sfnic) that attacks the fall armyworm, spodoptera frugiperda, survives as a mixture of nine genotypes (sfnic a to i) that all present genomic deletions, except variant b (complete genotype). sequencing of cloned restriction fragments revealed that genotypic variants lack between 5 and 16 of the open reading frames present in a contiguous sequence of 18 kb of the sfnic genome. the absence of oral infectivity of sfnic-c and -d variants is related to ... | 2005 | 16085811 |
| functional characterization of medaka cyp3a38 and cyp3a40: kinetics and catalysis by expression in a recombinant baculovirus system. | phylogenic analysis of the teleost genomic lineages has demonstrated the precedent for multiple genome duplications. among many of the genes duplicated, cytochrome p450 genes have undergone independent diversification, which can be traced to a single ancestral gene. in teleosts, cytochrome p450s, from all major families, have been identified. among these, the cyp3a family has been cloned in several teleost species and demonstrated to contain multiple paralogs differing in gene expression pattern ... | 2005 | 16112913 |
| insect growth regulatory effects of some extracts and sterols from myrtillocactus geometrizans (cactaceae) against spodoptera frugiperda and tenebrio molitor. | a methanol extract from the roots and aerial parts of myrtillocactus geometrizans (cactaceae) yielded peniocerol 1, macdougallin 2, and chichipegenin 3. the natural products 1, 2 their mixtures, meoh and ch(2)cl(2) extracts showed insecticidal and insect growth regulatory activity against fall armyworm [spodoptera frugiperda j. e. smith (lepidoptera: noctuidae)], an important insect pest of corn, and [tenebrio molitor (coleoptera)], a pest of stored grains in mexico. the most active compounds we ... | 2005 | 16122768 |
| a novel oscillating bioreactor bellocell: implications for insect cell culture and recombinant protein production. | bellocell is a novel packed bed bioreactor that allows alternating nutrient and gas transfer to a culture. spodoptera frugiperda sf-9 grown in the bellocell (300 ml culture) reached 1.3-1.5x10(7) cells ml-1 in 7-8 days and the total baculovirus-expressed protein yield was 2.3-times that in a stirred tank bioreactor (600 ml culture). the superior cell and protein yields underline the potential of bellocell for cell culture and recombinant protein production. | 2005 | 16132854 |
| spodoptera littoralis caspase-1, a lepidopteran effector caspase inducible by apoptotic signaling. | the baculovirus autographa californica multiple nucleopolyhedrovirus (acmnpv) can successfully infect spodoptera frugiperda sf9 cells, but in contrast, in spodoptera littoralis sl2 cells it induces apoptosis aborting the infection. to understand better the mechanism of induction and execution of apoptosis in sl2 cells, we identified and characterized the first spodoptera littoralis caspase, sl-caspase-1. sl-caspase-1 is an effector caspase that cleaves devd but not ietd and lehd substrates, and ... | 2005 | 16133869 |
| single cell manipulation, analytics, and label-free protein detection in microfluidic devices for systems nanobiology. | single cell analytics for proteomic analysis is considered a key method in the framework of systems nanobiology which allows a novel proteomics without being subjected to ensemble-averaging, cell-cycle, or cell-population effects. we are currently developing a single cell analytical method for protein fingerprinting combining a structured microfluidic device with latest optical laser technology for single cell manipulation (trapping and steering), free-solution electrophoretical protein separati ... | 2005 | 16152668 |
| acaricidal and cytotoxic activities of extracts from selected genera of australian lamiaceae. | crude foliar extracts of 67 species from six subfamilies of australian lamiaceae were screened by whole organism contact toxicity on the polyphagous mite tetranychus urticae koch (acari: tetranychidae) by using a potter precision spray tower. cytotoxicity assessments against insect cell lines from spodoptera frugiperda (j.e. smith) (lepidoptera: noctuidae) and drosophila melanogaster (meigen) (diptera: drosophilidae) also were made. the spodoptera cell line was more susceptible to extracts than ... | 2005 | 16156579 |
| diffusion dependent cell behavior in microenvironments. | understanding the interaction between soluble factors and cells in the cellular microenvironment is critical to understanding a wide range of diseases. microchannel culture systems provide a tool for separating diffusion and convection based transport making possible controlled studies of the effects of soluble factors in the cellular microenvironment. in this paper we compare the proliferation kinetics of cells in traditional culture flasks to those in microfluidic channels, and explore the rel ... | 2005 | 16175265 |
| visualization of inositol 1,4,5-trisphosphate receptor by atomic force microscopy. | inositol 1,4,5-trisphosphate (ip(3)) receptor (ip(3)r) acts as a ligand-gated channel that mediates neuronal signals by releasing ca(2+) from the endoplasmic reticulum. the three-dimensional (3d) structure of tetrameric ip(3)r has been demonstrated by using electron microscopy (em) with static specimens; however, the dynamic aspects of the ip(3)r structure have never been visualized in a native environment. here we attempt to measure the surface topography of ip(3)r in solution using atomic forc ... | 2006 | 16198054 |
| expression of slam (cdw150) on sf9 cell surface using recombinant baculovirus mediates measles virus infection in the nonpermissive cells. | signaling lymphocytic activation molecule (slam; also known as cdw150) has been reported as the receptor of measles virus (mv) interacting with mv hemagglutinin (mvh). in this study, we developed a baculovirus-derived vector, the bacmid-egfp, containing a reporter gene encoding the enhanced green fluorescent protein (egfp) under the control of the promoter of very late polyhedrin gene from autographa californica multiple nucleopolyhedrovirus (acmnpv), and employed the recombinant baculovirus to ... | 2005 | 16198138 |
| sequences of cdnas and expression of genes encoding chitin synthase and chitinase in the midgut of spodoptera frugiperda. | the focus of this study was on the characterization and expression of genes encoding enzymes responsible for the synthesis and degradation of chitin, chitin synthase (sfchsb) and chitinase (sfchi), respectively, in the midgut of the fall armyworm, spodoptera frugiperda. sequences of cdnas for sfchsb and sfchi were determined by amplification of overlapping pcr fragments and the expression patterns of these two genes were analyzed during insect development by rt-pcr. sfchsb encodes a protein of 1 ... | 2005 | 16203206 |
| visualization of a recombinant gene protein in the baculovirus expression vector system using confocal scanning laser microscopy. | expression of the recombinant protein beta-galactosidase in the spodoptera frugiperda sf-9 insect cell line infected by the autographa californica nuclear polyhedrosis virus expressing beta-galactosidase (acnpv-betagal) was visualized using confocal scanning laser microscopy with fluorescent staining of both the recombinant protein and the cell nucleus. the average size of the insect cells and the intracellular dna concentration both increased markedly, respectively reading 3.8- and 2.3-fold the ... | 1999 | 16232550 |
| significant increase in recombinant protein production of a virus-infected sf-9 insect cell culture of low moi under low dissolved oxygen conditions. | spodoptera frugiperda sf-9 insect cells were infected with recombinant autographa californica nuclear polyhedrosis virus at a low multiplicity of infection (moi) (0.1), and the effect of dissolved oxygen (do) on the production of a polyhedrin promoter-driven recombinant protein (beta-galactosidase), intrinsic proteases (carboxyl and cysteine proteases), and the virus was determined. the do concentrations used in the present study were 45%, 25%, 5%, and 1.3% of air saturation. at 5% do the cell g ... | 2002 | 16233329 |
| degradation of the s. frugiperda peritrophic matrix by an inducible maize cysteine protease. | a unique 33-kda cysteine protease (mir1-cp) rapidly accumulates at the feeding site in the whorls of maize (zea mays l.) lines that are resistant to herbivory by spodoptera frugiperda and other lepidopteran species. when larvae were reared on resistant plants, larval growth was reduced due to impaired nutrient utilization. scanning electron microscopy (sem) indicated that the peritrophic matrix (pm) was damaged when larvae fed on resistant plants or transgenic maize callus expressing mir1-cp. to ... | 2006 | 16243350 |
| effect of halothane on galphai-3 and its coupling to the m2 muscarinic receptor. | halothane is an effective bronchodilator and inhibits airway smooth muscle contraction in part by inhibiting intracellular signaling pathways activated by the m2 muscarinic receptor and its cognate inhibitory heterotrimeric guanosine-5'-triphosphate (gtp)-binding protein (g protein), gi. this study hypothesized that halothane inhibits nucleotide exchange at the alpha isoform-3 subunit of gi (galphai-3), but only when regulated by the m2 muscarinic receptor. | 2005 | 16249676 |
| similarity of cuticular lipids between a caterpillar and its host plant: a way to make prey undetectable for predatory ants? | ithomiine butterflies (nymphalidae) have long-lived, aposematic, chemically protected adults. however, little is known about the defense mechanisms in larvae and other juvenile stages. we showed that larvae mechanitis polymnia are defended from ants by a chemical similarity between their cuticular lipids and those of the host plant, solanum tabacifolium (solanaceae). this is a novel defense mechanism in phytophagous insects. a field survey during one season showed that larval survivorship was up ... | 2005 | 16273428 |
| development of a new high-throughput screening model for human high density lipoprotein receptor (cla-1) agonists. | to develop a new high-throughput screening model for human high-density lipoprotein (hdl) receptor (cd36 and limpii analogous-1, cla-1) agonists using cla-1-expressing insect cells. | 2005 | 16274109 |
| characterization of cathepsin l secreted by sf21 insect cells. | sf21 cells, derived from the spodoptera frugiperda pupa, are commonly used for the heterologous expression of proteins. while purifying recombinant proteins from this system we encountered a protease, secreted at high levels by sf21 cells, that readily degraded recombinant proteins and also tended to co-purify with histidine-tagged proteins from ni(2+) affinity columns. purification and characterization of the protease revealed that it has many properties consistent with cysteine proteases of th ... | 2005 | 16274658 |
| a homologue of cathepsin l identified in conditioned medium from sf9 insect cells. | gelatin zymography revealed the presence of proteolytic activity in conditioned medium (cm) from a serum-free, non-infected spodoptera frugiperda, sf9 insect cell culture. two peptidase bands at about 49 and 39 kda were detected and found to be proform and active form of the same enzyme. the 49-kda form was visible on zymogram gels in samples of cm taken on days 4 and 5 of an sf9 culture, while the 39-kda form was seen on days 6 and 7. on basis of the inhibitor profile and substrate range, the e ... | 2006 | 16283300 |
| studies of the intermediary metabolism in cultured cells of the insect spodoptera frugiperda using 13c- or 15n-labelled tracers. | insect cells can serve as host systems for the recombinant expression of eukaryotic proteins. using this platform, the controlled expression of 15n/13c labelled proteins requires the analysis of incorporation paths and rates of isotope-labelled precursors present in the medium into amino acids. for this purpose, spodoptera frugiperda cells were grown in a complex medium containing [u-13c6]glucose. in a second experiment, cultures of s. frugiperda were grown in the presence of 15n-phenylalanine. | 2005 | 16285881 |
| casein kinase 2 during early embryonic development in silkworm bombyx mori: cdna sequence, gene expression, and enzyme activity. | to clarify the property of casein kinase 2 (ck2) during early embryonic development in the silkworm, we compared the phosphorylation activities of ck2 in non-diapause and diapause eggs until 60 h after oviposition. in nondiapause eggs, the phosphorylated signals were found at each stage and became progressively stronger through each stage. on the other hand, in diapause eggs, the strongest phosphorylated signals were found at approximately 12 to 24 h after oviposition and became progressively we ... | 2005 | 16287624 |
| expression and characterization of human vascular endothelial growth factor (vegf165) in insect cells. | vascular endothelial growth factor (vegf) is the best characterized multifunctional protein which plays a key role in normal and pathologic angiogenesis. the gene encoding the human vegf165 was cloned from the ovarian carcinoma cell line (ovcar3) and expressed in insect cells using the baculovirus expression vector system. the recombinant human vegf165 (rhvegf165) protein produced by sf21 (spodoptera frugiperda) cells underwent a similar processing compared with mammalian cells, including effici ... | 2006 | 16289914 |