| impact of bt cottons expressing one or two insecticidal proteins of bacillus thuringiensis berliner on growth and survival of noctuid (lepidoptera) larvae. | a series of laboratory assays were performed to compare the relative impact of commercial and experimental cultivars of cotton, gossypium hirsutum (l.), expressing zero, one, or two insecticidal proteins of bacillus thuringiensis berliner, on several lepidopteran pests. assays in which larvae were fed fresh plant tissue indicated that dual-toxin b. thuringiensis (bt) cultivars, expressing both cry1ac and cry2ab endotoxins of b. thuringiensis, were more toxic to bollworms, helicoverpa zea (boddie ... | 2001 | 11425033 |
| antifeedant constituents from fagara macrophylla. | analysis of the polar fractions of an etoh extract obtained from the bark of the african medicinal plant fagara macrophylla led to the isolation and identification of the alkaloids oblongine (6), tembetarine (7) and magnoflorine (8) and the flavonoid hesperidin (9). these compounds, together with other metabolites (1--5) previously isolated from f. macrophylla, were tested for antifeedant activity in a binary-choice bioassay. the acridone alkaloid xanthoxoline (4) was found to have a potent anti ... | 2001 | 11429249 |
| effect of baculovirus infection on the mrna and protein levels of the spodoptera frugiperda eukaryotic initiation factor 4e. | the cdna sequence of eukaryotic translation initiation factor eif4e was derived from a spodoptera frugiperda cdna library. eight tryptophan residues, typical for eif4e, are strictly conserved in the encoded 210 amino acid protein. a polyclonal antiserum detected a 26 kda protein in lepidopteran cell lines, but not in dipteran cells. sf21 cells have a single eif4e gene copy, which is transcribed into a 1500 nt transcript. infection with acmnpv resulted in a decrease in eif4e mrna starting between ... | 2001 | 11437917 |
| characterization of lysosomal acid lipase mutations in the signal peptide and mature polypeptide region causing wolman disease. | wolman disease results from an inherited deficiency of lysosomal acid lipase (lal; ec 3.1.1.13). this enzyme is essential for the hydrolysis of cholesteryl esters and triacylglycerols derived from endocytosed lipoproteins. because of a complete absence of lal activity, wolman patients accumulate progressive amounts of cholesteryl esters and triacylglycerols in affected tissues. to investigate the nature of the genetic defects causing this disease, mutations in the lal gene from three subjects of ... | 2001 | 11441129 |
| cannabinoid receptors are absent in insects. | the endocannabinoid system exerts an important neuromodulatory role in mammals. knockout mice lacking cannabinoid (cb) receptors exhibit significant morbidity. the endocannabinoid system also appears to be phylogenetically ancient--it occurs in mammals, birds, amphibians, fish, sea urchins, leeches, mussels, and even the most primitive animal with a nerve network, the hydra. the presence of cb receptors, however, has not been examined in terrestrial invertebrates (or any member of the ecdysozoa) ... | 2001 | 11447587 |
| antibody-secreting cell responses to rotavirus proteins in gnotobiotic pigs inoculated with attenuated or virulent human rotavirus. | because of their similarities to infants in mucosal immune responses and their susceptibility to human rotavirus (hrv) diarrhea, gnotobiotic pigs provide a useful model for rotaviral disease. in this study, we performed quantitative enzyme-linked immunospot (elispot) assays to measure local and systemic isotype-specific antibody-secreting cell (asc) responses to individual structural (vp4, vp6, and vp7) and nonstructural (nsp3 and nsp4) proteins of wa hrv. the spodoptera frugiperda cells express ... | 2001 | 11473996 |
| expression of c1 esterase inhibitor by the baculovirus expression vector system: preparation, purification, and characterization. | c1 esterase inhibitor (c1inh) is an important regulator of the classical complement pathway. hereditary deficiency of c1inh causes angioedema of the skin, gut, and respiratory tissues that may be fatal. c1inh replacement therapy may be lifesaving for patients with this disorder. the objective of this study was to evaluate the use of the baculovirus expression vector system for mass producing biologically active human recombinant (rc1inh). a recombinant baculovirus was constructed coding the huma ... | 2001 | 11483003 |
| comparison of growth and recombinant protein expression in two different insect cell lines in attached and suspension culture. | culture conditions required for obtaining maximum recombinant protein concentrations from two cell lines, spodoptera frugiperda (iplbeta-sf21-ae) and trichoplusia ni (tn 5beta-1-4), were determined in this work. conditions studied include mode of culture (suspended vs attached), agitation rates, inoculum sizes, cell concentration at the time of infection, and various serum-free media (sfm). results were compared with the performance of attached cultures in tnm-fh with 10% fetal bovine serum. gro ... | 2001 | 11485429 |
| an arabidopsis inositol phospholipid kinase strongly expressed in procambial cells: synthesis of ptdins(4,5)p2 and ptdins(3,4,5)p3 in insect cells by 5-phosphorylation of precursors. | we have cloned a phosphatidylinositol-4-phosphate 5-kinase (pip5k) cdna (atp5k1) from arabidopsis thaliana. by the application of cell permeabilization and short-term nonequilibrium labelling we show that expression of atp5k1 in baculovirus-infected insect (spodoptera frugiperda) cells directs synthesis of ptdins(4,5)p2 and ptdins(3,4,5)p3. the same phosphoinositides were produced by isolated whole-cell membrane fractions of atp5k1-expressing insect cells. their synthesis was not affected by add ... | 2001 | 11489170 |
| co-expression of human cytochrome p4501a1 (cyp1a1) variants and human nadph-cytochrome p450 reductase in the baculovirus/insect cell system. | 1. three human cytochrome p4501a1 (cyp1a1) variants, wild-type (cyp1a1.1), cyp1a1.2 (1462v) and cyp1a1.4 (t461n), were co-expressed with human nadph-p450 reductase (or) in spodoptera frugiperda (sf9) insect cells by baculovirus co-infection to elaborate a suitable system for studying the role of cypa1 polymorphism in the metabolism of exogenous and endogenous substrates. 2. a wide range of conditions was examined to optimize co-expression with regard to such parameters as relative multiplicity o ... | 2001 | 11513247 |
| a study of the autographa californica multiple nucleopolyhedrovirus odv envelope protein p74 using a gfp tag. | the autographa californica multiple nucleopolyhedrovirus (acmnpv) protein p74 is associated with the occlusion-derived virus (odv) envelope. p74 is essential for oral infectivity of odv and has been proposed to play a role in midgut attachment and/or fusion. in this study, p74 protein was expressed in-frame with green fluorescent protein (gfp) to create a p74-gfp chimera. the c-terminal gfp portion of the chimera facilitated visualization of the trafficking of p74 in baculovirus-infected spodopt ... | 2001 | 11514740 |
| establishment and characterization of insect cell lines from 10 lepidopteran species. | cell lines from selected lepidopteran species were established for the overall purpose of use in baculovirus production. a total of 36 new cell lines from 10 lepidopteran species were generated, including cell lines from a pyralid, the european corn borer, ostrinia nubilalis, a plutellid, the diamondback moth, plutella xylostella, as well as eight noctuids: the black cutworm, agrotis ipsilon, the celery looper, anagrapha falcifera, the velvetbean caterpillar, anticarsia gemmatalis, the corn earw ... | 2001 | 11515970 |
| insect growth regulator and insecticidal activity of beta-dihydroagarofurans from maytenus spp. (celastraceae). | from the aerial parts of maytenus disticha, we have isolated 9beta-benzoyloxy-1alpha,2alpha,6beta,8alpha,15penta-acetoxy-dihydro-beta-agarofuran (1) and from seeds of maytenus boaria 9beta-furoyloxy-1alpha,6beta,8alpha-triacetoxy-dihydro-beta-agarofuran (2). these compounds and their meoh and hexane/ethyl acetate (1:1 v/v) extracts were evaluated for their effects on the fall armyworm (spodoptera frugiperda). toosendanin, a commercial insecticide derived from melia azedarach was used as a positi ... | 2001 | 11531097 |
| effect of moi ratio on the composition and yield of chimeric infectious bursal disease virus-like particles by baculovirus co-infection: deterministic predictions and experimental results. | virus-like particles (vlps) are empty particles consisting of virus capsid proteins that closely resemble native virus but are devoid of the native viral nucleic acids and therefore have attracted significant attention as noninfectious vaccines. a recombinant baculovirus, vibd-7, which encodes the structural proteins (vp2, vp3, and vp4) of infectious bursal disease virus (ibdv), produces native ibd vlps in infected spodoptera frugiperda insect cells. another baculovirus, vedlh-22, encodes vp2 th ... | 2001 | 11536133 |
| influence of simulated microgravity on the longevity of insect-cell culture. | simulated microgravity within the nasa high aspect rotating-wall vessel (harv) provides a quiescent environment to culture fragile insect cells. in this vessel, the duration of stationary and death phase for cultures of spodoptera frugiperda cells was greatly extended over that achieved in shaker-flask controls. for both harv and control cultures, s. frugiperda cells grew to concentrations in excess of 1 x 10(7) viable cells ml-1 with viabilities greater than 90%. in the harv, stationary phase w ... | 1997 | 11536801 |
| toxicity of pyrethroids and effect of synergists to larval and adult helicoverpa zea, spodoptera frugiperda, and agrotis ipsilon (lepidoptera: noctuidae). | based on 48 h ld50 estimates from topical bioassays, cypermethrin was more toxic than permethrin to helicoverpa zea (boddie) larvae and adults; however, the two pyrethroids did not differ significantly in their relative toxicities to spodoptera frugiperda (j. e. smith) and agrotis ipsilon (hufnagle). larvae of each species generally were more susceptible to cypermethrin and permethrin than respective adults. the only exception to this generalization occurred with h. zea where slight overlap of t ... | 2001 | 11561845 |
| pharmacokinetic mechanisms associated with synergism by def of cypermethrin toxicity in larval and adult helicoverpa zea, spodoptera frugiperda, and agrotis ipsilon (lepidoptera: noctuidae). | penetration, metabolism, and excretion of radiocarbon were observed after topical treatment of helicoverpa zea (boddie), spodoptera frugiperda (j. e. smith), and agrotis ipsilon (hufnagle) larvae and adults with cypermethrin-14c. these pharmacokinetic events usually were higher with trans-cypermethrin-14c than with cis-cypermethrin-14c. they also were generally higher with h. zea and s. frugiperda than with a. ipsilon, and they were higher in larvae than in adults. no marked sex differences in t ... | 2001 | 11561846 |
| def sensitive esterases in homogenates of larval and adult helicoverpa zea, spodoptera frugiperda, and agrotis ipsilon (lepidoptera: noctuidae). | homogenates of helicoverpa zea (boddie), agrotis ipsilon (hufnagle), and spodoptera frugiperda (j. e. smith) third instars and adults contained s,s,s-tri-n-butyl phosphorotrithioate (def)-sensitive enzymes that hydrolyzed trans-cypermethrin and two known esterase substrates, alpha-naphthyl acetate and beta-naphthyl acetate. except for h. zea with alpha-naphthyl acetate, larval preparations were more active than those of adults, and no marked sex differences were apparent. the hydrolysis of trans ... | 2001 | 11561847 |
| measurement of membrane fusion activity from viral membrane fusion proteins based on a fusion-dependent promoter induction system in insect cells. | a number of viral membrane fusion proteins can be expressed alone on the surface of host cells, and then triggered to induce cell-to-cell fusion or syncytium formation. although rapid and easily observed, syncytium formation is not easily quantified and differences in fusion activity are not easily distinguished or measured. to address this problem, we developed a rapid and quantitative cell-to-cell fusion system that is useful for comparative analysis and may be suitable for high throughput scr ... | 2001 | 11562545 |
| a comparison of techniques for detecting invertebrate iridescent virus 6. | the aim of this study was to compare the sensitivity and precision of various methods for the detection and quantification of invertebrate iridescent virus 6 (iiv-6) (iridoviridae) isolated from a the stem-boring moth chilo suppressalis, and to apply these techniques to the detection of covert infections in the wax moth, galleria mellonella. the relationship between the virus concentration and absorbance at 260 nm was linear over the range of 1.6 x 10(9)-5.6 x 10(10) particles/ml. tcid(50) assay ... | 2001 | 11576637 |
| processing of cry1ab delta-endotoxin from bacillus thuringiensis by manduca sexta and spodoptera frugiperda midgut proteases: role in protoxin activation and toxin inactivation. | activation of cry protoxins is carried out by midgut proteases. this process is important for toxicity and in some cases for specificity. commercial proteases have been used for in vitro protoxin activation. in the case of cry1a protoxins, trypsin digestion generates a toxic fragment of 60-65 kda. here, we have analyzed the in vitro and in vivo activation of cry1ab. we found differences in the processing of cry1ab protoxin by manduca sexta and spodoptera frugiperda midgut proteases as compared t ... | 2001 | 11583928 |
| investigation of sequential behavior of carboxyl protease and cysteine protease activities in virus-infected sf-9 insect cell culture by inhibition assay. | proteases produced during the culture of spodoptera frugiperda sf-9 cells infected with autographa californica nuclear polyhedrosis virus (acnpv) were assayed with various protease inhibitors. this inhibitory analysis revealed that: (1) carboxyl and cysteine proteases were predominantly produced by the insect cells infected with recombinant acnpv, the gene of which encoded a variant of green fluorescent protein in a portion of the polyhedrin gene of the baculovirus, and (2) the protease activity ... | 2001 | 11601624 |
| expression and purification of manduca sexta prophenoloxidase-activating proteinase precursor (propap) from baculovirus-infected insect cells. | analogous to human thrombin, prophenoloxidase-activating proteinase (pap) is a terminal enzyme of a serine proteinase cascade in the tobacco hornworm manduca sexta. in order to purify and study the activating enzyme for pap from this insect, we produced the zymogen of pap (propap) in a bacterial expression system. the affinity-purified protein was then used as an antigen to generate a specific rabbit antiserum. immunoblot analysis indicated that the propap was present at a low level in manduca l ... | 2001 | 11676609 |
| deletions in the ac-iap1 gene of the baculovirus acmnpv occur spontaneously during serial passage and confer a cell line-specific replication advantage. | the psti-i region of the autographa californica multicapsid nucleopolyhedrovirus (acmnpv) genome was previously shown to be a frequent target of spontaneous deletions during serial virus passage in tn-368 cells (kumar and miller, virus res. 7 (1987) 335). analysis of two of these serial passage mutants showed that a portion of the ac-iap1 gene was deleted. to directly test the effect of loss of ac-iap1, three different deletions in ac-iap1 were introduced into recombinant viruses and the ability ... | 2001 | 11682127 |
| recovery of infectious pariacoto virus from cdna clones and identification of susceptible cell lines. | pariacoto virus (pav) is a nodavirus that was recently isolated in peru from the southern armyworm, spodoptera eridania. virus particles are non enveloped and about 30 nm in diameter and have t=3 icosahedral symmetry. the 3.0-a crystal structure shows that about 35% of the genomic rna is icosahedrally ordered, with the rna forming a dodecahedral cage of 25-nucleotide (nt) duplexes that underlie the inner surface of the capsid. the pav genome comprises two single-stranded, positive-sense rnas: rn ... | 2001 | 11711613 |
| interaction of lyssaviruses with the low-affinity nerve-growth factor receptor p75ntr. | the low-affinity nerve-growth factor receptor p75ntr interacts in vitro with the rabies virus (rv) glycoprotein and serves as a receptor for rv. the lyssavirus genus comprises seven genotypes (gts) of rabies and rabies-related viruses. the ability of p75ntr to interact with the glycoprotein of representative lyssaviruses from each gt was investigated. this investigation was based on a specific binding assay between bsr cells infected with a lyssavirus and spodoptera frugiperda (sf21) cells expre ... | 2001 | 11714960 |
| unfolding affects insect cell permeabilization by bacillus thuringiensis cry1c toxin. | bacillus thuringiensis cry toxins are efficient, environment-friendly biological insecticides. their molecular mode of action on target insect cells remains largely unknown. the aim of this study was to investigate the relation between the conformational state of the cry1c toxin and its ionophoric activity on live sf9 cells of spodoptera frugiperda, a target insect for this protein. potassium ion movement induced by cry1c across the cell membrane was measured with a fluorescent assay developed p ... | 2001 | 11718667 |
| evidence of oxidative stress following the viral infection of two lepidopteran insect cell lines. | the infection of spodoptera frugiperda sf-9 (sf-9) and trichoplusia ni bti-tn-5b1-4 (tn-5b1-4) insect cell lines with autographa californica multiple nucleopolyhedrovirus (acmnpv) resulted in increased levels of lipid hydroperoxides and protein carbonyls. in addition, the viral infection resulted in a significant decrease in the reduced glutathione to oxidized glutathione (2gsh/gssg) ratio. these results are all consistent with an increased level of oxidative stress as a result of the viral infe ... | 2001 | 11728817 |
| (-)s amisulpride binds with high affinity to cloned dopamine d(3) and d(2) receptors. | amisulpride is a substituted benzamide antipsychotic with nanomolar affinity and high selectivity for dopamine d(2) and dopamine d(3) receptors. the interaction of racemic (+/-)rs amisulpride and its two enantiomers (+)r and (-)s with dopamine d(2) and dopamine d(3) receptors subtypes were compared with that of haloperidol. binding studies were performed using either [3h]spiperone or [3h]nemonapride in baculovirus/spodoptera frugiperda insect (sf-9) cell system expressing either the human dopami ... | 2001 | 11740949 |
| field efficacy and seasonal expression profiles for terminal leaves of single and double bacillus thuringiensis toxin cotton genotypes. | examination of commercial cry1ac transgenic bacillus thuringiensis berliner (bt) cotton varieties (bollgard, monsanto, st. louis, mo) and an experimental cry1ac + cry2ab transgenic bt cotton variety (bollgard ii, monsanto) for lepidopteran field efficacy was conducted during the 2000 growing season. in addition, a commercially available (envirologix, portland, me) quantification assay (elisa) was used to measure and profile the expression levels of cry proteins in two of these varieties ['dp50b, ... | 2001 | 11777069 |
| dual signalling by different octopamine receptors converges on adenylate cyclase in sf9 cells. | the sf9 insect cell line, derived from spodoptera frugiperda, was used to study the regulation of adenylate cyclase (ac) activity by octopamine receptors. the cyclic amp (camp) production was stimulated in a concentration-dependent manner by octopamine. octopamine also elicited a rise in cytosolic ca(2+). the ca(2+) elevation was independent of the camp elevation whereas the camp elevation was partially inhibited by removal of ca(2+). the antagonistic effects of a series of compounds were tested ... | 2002 | 11804800 |
| assessment of foreign protein production by recombinant heliothis (helicoverpa) armigera entomopoxviruses in spodoptera frugiperda cells. | this report describes the first production of recombinant forms of heliothis (helicoverpa) armigera entomopoxvirus (haepv). these haepvs are engineered at either the spheroidin or fusolin locus, to produce the green fluorescent marker protein (gfp). the growth properties of these recombinant haepvs, in comparison to the parental haepv, were assessed in cultured spodoptera frugiperda sf9 cells. additionally, gfp production by these recombinant haepvs was compared to that of a gfp-expressing recom ... | 2002 | 11807239 |
| 5-ht-receptor-induced changes of the intracellular camp level monitored by a hyperpolarization-activated cation channel. | the hvcng channel from the moth heliothis virescens is highly sensitive to camp concentrations ranging between 0.1 microm and 5 microm. this hvcng channel was over-expressed in spodoptera frugiperda (sf.9) cells to measure endogenous camp levels. hyperpolarization-activated inward currents were measured in the whole-cell patch-clamp configuration with pipettes filled with different camp concentrations to calibrate the system. varying the camp concentration between 0 microm and 100 microm in the ... | 2002 | 11810212 |
| penetration of cuticle and proliferation in hemolymph by paecilomyces fumosoroseus isolates that differ in virulence against lepidopteran larvae. | frequencies of cuticular penetration and speed of proliferation in hemolymph were demonstrated for two isolates of paecilomyces fumosoroseus that differ in virulence against diamondback moth, plutella xylostella. penetrant hyphae of virulent isolate 4461 were visible in larval cuticle cross-sections of diamondback moth and fall armyworm, spodoptera frugiperda, within 22 h after inoculation. virtually no penetration was observed for isolate 1576 for up to 52 h after inoculation. isolate 4461 also ... | 2001 | 11812110 |
| comparison of blastospores of two paecilomyces fumosoroseus isolates: in vitro traits and virulence when injected into fall armyworm, spodoptera frugiperda. | blastospores of two isolates of paecilomyces fumosoroseus were compared to determine their productivity in vitro and their relative pathogenicity after injection of fall armyworm, spodoptera frugiperda. blastospores of less virulent p. fumosoroseus isolate 1576 are smaller than those of isolate 4461, and they germinate and proliferate more slowly in vitro. the pathogenicity of injected blastospores of isolate 1576 against s. frugiperda varies with larval size. in small larvae, percentage mortali ... | 2001 | 11812120 |
| characterization of wolbachia host cell range via the in vitro establishment of infections. | maternally transmitted bacteria of the genus wolbachia are obligate, intracellular symbionts that are frequently found in insects and cause a diverse array of reproductive manipulations, including cytoplasmic incompatibility, male killing, parthenogenesis, and feminization. despite the existence of a broad range of scientific interest, many aspects of wolbachia research have been limited to laboratories with insect-rearing facilities. the inability to culture these bacteria outside of the invert ... | 2002 | 11823204 |
| rat monoclonal antibodies differentiating between the epstein-barr virus nuclear antigens 2a (ebna2a) and 2b (ebna2b). | rat monoclonal antibodies were produced against the c-terminus of epstein-barr virus nuclear antigens 2a (ebna2a) and 2b (ebna2b) expressed as bacterial trpe fusion proteins. the initial screening was performed using a soluble bacterial extract containing the fusion proteins. positive hybridomas were confirmed by immunofluorescence on sf158 (spodoptera frugiperda) insect cells infected with recombinant baculovirus (autographa californica nuclear polyhedrosis virus) and expressing the complete eb ... | 1995 | 11831716 |
| the human histamine h2-receptor couples more efficiently to sf9 insect cell gs-proteins than to insect cell gq-proteins: limitations of sf9 cells for the analysis of receptor/gq-protein coupling. | the human histamine h2-receptor (hh2r) couples to gs-proteins to activate adenylyl cyclase and to gq-proteins to activate phospholipase c, but phospholipase c activation has not consistently been observed. the aim of this study was to compare coupling of hh2r to insect and mammalian gs- and gq-proteins in spodoptera frugiperda (sf9) cells. interaction of hh2r with mammalian g proteins was assessed with coexpressed proteins or receptor-galpha fusion proteins that enhance coupling efficiency. hh2r ... | 2002 | 11841575 |
| heterologous expression and topography of the main intrinsic protein (mip) from rat lens. | wild type rat lens main intrinsic protein (mip) and mip mutated (f73i, f75l) to resemble the glycerol facilitator of escherichia coli in the region of the npa1 box were used to investigate the topology of mip in the membrane of spodoptera frugiperda (sf21) cells using the baculovirus expression system and expression in mouse erythroid leukaemia cells (mel c88). differential fixation for staining was used, with paraformaldehyde for externally exposed antigenic sites, and acetone for both external ... | 2002 | 11852078 |
| ph-dependent channel activity of heterologously-expressed main intrinsic protein (mip) from rat lens. | wild-type rat lens main intrinsic protein (mip) was heterologously expressed in the membrane of spodoptera frugiperda (sf21) cells using the baculovirus expression system and in mouse erythroid leukaemia cells (mel c88). both mel and sf21 cell lines expressing wild-type mip were investigated for the conductance of ions using a whole cell patch clamp technique. an increase in conductance was seen in both expression systems, particularly on lowering the ph to 6.3. in sf21 cells, addition of antibo ... | 2002 | 11852079 |
| kinetic properties of bifunctional 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase from spinach leaves. | a cdna encoding 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase was isolated from a spinacia oleracea leaf library and used to express a recombinant enzyme in escherichia coli and spodoptera frugiperda cells. the insoluble protein expressed in e. coli was purified and used to raise antibodies. western blot analysis of a protein extract from spinach leaf showed a single band of 90.8 kda. soluble protein was purified to homogeneity from s. frugiperda cells infected with recombinant baculoviru ... | 2002 | 11856361 |
| diversity of bacillus thuringiensis strains from colombia with insecticidal activity against spodoptera frugiperda (lepidoptera:noctuidae). | to identify and characterize bacillus thuringiensis strains highly toxic to spodoptera frugiperda, and to explore the genetic diversity of such strains. | 2002 | 11872122 |
| trypanosoma evansi: cloning and expression in spodoptera frugiperda [correction of fugiperda] insect cells of the diagnostic antigen rotat1.2. | a complementary dna encoding the variant surface glycoprotein (vsg) of trypanosoma evansi rode trypanozoon antigenic type (rotat)1.2, currently used for experimental serological diagnosis of t. evansi infection in livestock, was cloned as a recombinant plasmid and sequenced. a recombinant baculovirus containing the coding region of rotat1.2 vsg was constructed to express the protein in spodoptera frugiperda [corrected] insect cells. from this, sufficient quantities of the recombinant protein are ... | 2001 | 11888244 |
| production and partial characterization of mouse monoclonal antibodies recognizing common cytokine receptor gamma chain (gammac) of human, mouse and primate origin. | monoclonal antibodies specific for the common cytokine receptor gamma chain, gammac, were produced using traditional hybridoma technology. fusion of p3x63-ag8.653 myeloma cells with splenocytes from balb/c mice immunized with spodoptera frugiperda insect cells infected with the recombinant baculovirus vl1392-hil-2rgamma resulted in several hybridoma cell clones producing monoclonal gammac-specific antibodies. four of these antibody-producing clones, iiic3, iiie8, ig3 and if10c5, were further cha ... | 2001 | 11890567 |
| expression of winged bean basic agglutinin in spodoptera frugiperda insect cell expression system. | in this paper we report the successful expression of the winged bean basic agglutinin (wba i) in insect cells infected with a recombinant baculovirus carrying the wba i gene and its characterization in terms of its carbohydrate binding properties. the expressed protein appears to have a lower molecular weight than the native counterpart which is consistent with the lack of glycosylation of the former. moreover, the expressed protein maintains its dimeric nature. hence, a role for glycosylation i ... | 2001 | 11893002 |
| expression of the human mu opioid receptor in a stable sf9 cell line. | the cdna that encodes the human mu opioid receptor (hmor) has been cloned and expressed in spodoptera frugiperda (sf9) cells using a nonlytic vector system. the coding sequence fused to the cleavable glycoprotein signal peptide gp 64, and a c-terminal histidine tag was placed under the transcriptional control of the orgyia pseudotsugata multicapsid nucleopolyhedrosis virus immediate-early 2 (opie2) promoter. transfected cells were selected using zeocin resistance and the receptor was constitutiv ... | 2002 | 11911927 |
| functional characterization of the type 1 inositol 1,4,5-trisphosphate receptor coupling domain sii(+/-) splice variants and the opisthotonos mutant form. | the type 1 inositol (1,4,5)-trisphosphate receptor (insp3r1) plays a critical role in ca2+ signaling in cells. neuronal and nonneuronal isoforms of the insp3r1 differ by alternative splicing in the coupling domain of the insp3r1 (sii site). deletion of 107 amino acids from the coupling domain of the insp3r1 results in epileptic-like behaviors in opisthotonos (opt) spontaneous mouse mutant. using spodoptera frugiperda cells expression system, we compared single-channel behavior of recombinant ins ... | 2002 | 11916857 |
| 5-hydroxytryptamine 4(a) receptor is coupled to the galpha subunit of heterotrimeric g13 protein. | serotonin (5-hydroxytryptamine (5-ht)) is an important neurotransmitter that regulates multiple events in the central nervous system. many of the 5-ht functions are mediated via g protein-coupled receptors that are coupled to multiple heterotrimeric g proteins, including g(s), g(i), and g(q) subfamilies (martin, g. r., eglen, r. m., hamblin, m. w., hoyer, d., and yocca, f. (1998) trends pharmacol. sci. 19, 2-4). here we show for the first time that the 5-hydroxytryptamine 4(a) receptor (5-ht(4(a ... | 2002 | 11923294 |
| fluorometric microplate assay to measure glutathione s-transferase activity in insects and mites using monochlorobimane. | elevated levels of glutathione s-transferases (gsts) play a major role as a mechanism of resistance to insecticides and acaricides in resistant pest insects and mites, respectively. such compounds are either detoxicated directly via phase i metabolism or detoxicated by phase ii metabolism of metabolites as formed by microsomal monooxygenases. here we used monochlorobimane (mcb) as an artificial substrate and glutathione to determine total gst activity in equivalents of single pest insects and sp ... | 2002 | 11950219 |
| biochemical properties of purified recombinant human beta-carotene 15,15'-monooxygenase. | beta-carotene 15,15'-monooxygenase (bco), formerly known as beta-carotene 15,15'-dioxygenase, catalyzes the first step in the synthesis of vitamin a from dietary carotenoids. we have biochemically and enzymologically characterized the purified recombinant human bco enzyme. a highly active bco enzyme was expressed and purified to homogeneity from baculovirus-infected spodoptera frugiperda 9 insect cells. the k(m) and v(max) of the enzyme for beta-carotene were 7 microm and 10 nmol retinal/mg x mi ... | 2002 | 11960992 |
| the high-affinity calcium[bond]calmodulin-binding site does not play a role in the modulation of type 1 inositol 1,4,5-trisphosphate receptor function by calcium and calmodulin. | modulation of the inositol 1,4,5-trisphosphate (insp(3)) receptors (insp(3)r) by cytosolic calcium (ca(2+)) plays an essential role in ca(2+) signalling, but structural determinants and mechanisms responsible for the insp(3)r regulation by ca(2+) are poorly understood. in the present study, we expressed rat insp(3)r type 1 (insp(3)r1) in spodoptera frugiperda cells using a baculovirus-expression system and reconstituted the recombinant insp(3)r1 into planar lipid bilayers for functional analysis ... | 2002 | 11972451 |
| purification and biochemical characterization of some of the properties of recombinant human kynureninase. | recombinant human kynureninase (l-kynurenine hydrolase, ec 3.7.1.3) was purified to homogeneity (60-fold) from spodoptera frugiperda (sf9) cells infected with baculovirus containing the kynureninase gene. the purification protocol comprised ammonium sulfate precipitation and several chromatographic steps, including deae-sepharose cl-6b, hydroxyapatite, strong anionic and cationic separations. the purity of the enzyme was determined by sds/page, and the molecular mass verified by maldi-tof ms. th ... | 2002 | 11985583 |
| juvenile hormone biosynthesis in the fall armyworm, spodoptera frugiperda (lepidoptera, noctuidae). | the in vitro production of juvenile hormones (jh) was investigated by using corpora allata (ca) of larvae and corpora cardiaca-corpora allata (cc-ca) complexes of adult females of the fall armyworm spodoptera frugiperda. in female moths, jh release was high compared to that in 5th and 6th instar larvae. concentrations of 0.11-0.12 mm methionine, 180-200 mm na(+), 5.8-8.3 mm k(+), 10-50 mm ca(2+) and a ph range of 5.7-6.3 yielded optimal incorporation of l-[methyl-(3)h] methionine in vitro by cc- ... | 2002 | 11997221 |
| biological activity of astilbin from dimorphandra mollis against anticarsia gemmatalis and spodoptera frugiperda. | astilbin was isolated in high yield from dimorphandra mollis, and its insecticidal and growth inhibiting activity by stomach ingestion were evaluated against anticarsia gemmatalis and spodoptera frugiperda. the insecticidal activity of astilbin, the weight reduction of the larval phase and the prolongation of the larval and pupal phases were verified for both species. astilbin was identified on the base of its nmr, ms and physical data. | 2002 | 11997979 |
| grass species and endophyte effects on survival and development of fall armyworm (lepidoptera: noctuidae). | grass selections including 10 zoysiagrasses, 18 paspalums, 34 bermuda grasses, tall fescue, creeping red fescue, and perennial ryegrasses with and without endophyte were evaluated for potential resistance to fall armyworm, spodoptera frugiperda (j. e. smith), larvae. laboratory evaluations assessed the degree of antibiosis among >70 grass lines to first-instar fall armvworms. when all parameters measured were considered, the trend in resistance to fall armyworm among endophyte-infected (e+) and ... | 2002 | 12020031 |
| molecular cloning and induction of nuclear receptors from insect cell lines. | fragments of ecr and usp were cloned from two insect cell lines, sf21 and high five cells (derived respectively from spodoptera frugiperda and trichoplusia ni), using a pcr-based approach employing degenerate primers designed on the basis of conserved regions of nuclear receptors, together with 5'- and 3'-race. an additional orphan nuclear receptor, hr4 fragment, was cloned from high five cells. comparison of these fragments with manduca sexta counterparts showed that the cloned sfecr [ecdysone ... | 2002 | 12020840 |
| identification of protein tyrosine phosphatase 1b and casein as substrates for 124-v-mos. | the mos proto-oncogene encodes a cytoplasmic serine/threonine-specific protein kinase with crucial function during meiotic cell division in vertebrates. based on oncogenic amino acid substitutions the viral derivative, 124-v-mos, displays constitutive protein kinase activity and functions independent of unknown upstream effectors of mos protein kinase. we have utilized this property of 124-v-mos and screened for novel mos substrates in immunocomplex kinase assays in vitro. | 2002 | 12022922 |
| expression of the movement protein of tomato spotted wilt virus in its insect vector frankliniella occidentalis. | tomato spotted wilt virus (tswv) is able to infect both its botanical hosts and its insect vector (thrips). in plant tissue the ns(m) protein of tswv functions as viral movement protein (mp), aggregating into plasmodesma-penetrating tubules to establish cell-to-cell movement. as upon heterologous expression ns(m) was able to form similar tubules on the surface of insect (spodoptera frugiperda) cells, we have now investigated the expression and cellular manifestation of this protein in infected t ... | 2002 | 12038691 |
| chromosome instability in spodoptera frugiperda sf-9 cell line. | homogeneous cell lines are essential in industry and research if reliable and reproducible data are to be obtained. the spodoptera frugiperda (sf-9) cell line routinely used for the production of recombinant proteins was found to be heterogeneous, containing a mixture of diploid and tetraploid cells. using dilution-cloning techniques, diploid and tetraploid subpopulations were isolated from a sf-9 parental cell line, and their cytogenetic state was monitored using vinblastine to arrest cells in ... | 2002 | 12052082 |
| optimization of baculovirus-mediated expression and purification of hexahistidine-tagged murine dna (cytosine-c5)-methyltransferase-1 in spodoptera frugiperda 9 cells. | enzymatic dna methylation of carbon 5 of cytosines is an epigenetic modification that plays a role in regulating gene expression, differentiation, and tumorigenesis. dna (cytosine-c5)-methyltransferase-1 is the enzyme responsible for maintaining established methylation patterns during replication in mammalian cells. it is composed of a large ( approximately 1100 amino acids (a.a.)) amino-terminal region containing many putative regulatory domains and a smaller ( approximately 500 a.a.) carboxy-t ... | 2002 | 12071696 |
| lack of involvement of haemocytes in the establishment and spread of infection in spodoptera frugiperda larvae infected with the baculovirus autographa californica m nucleopolyhedrovirus by intrahaemocoelic injection. | it is thought that insect haemocytes, or blood cells, play an important role in baculovirus pathogenesis by amplifying and helping to spread the infection within the insect. here, infection is described of the lepidopteran noctuid species spodoptera frugiperda with the baculovirus autographa californica m nucleopolyhedrovirus (acmnpv). late instar s. frugiperda larvae were infected by intrahaemocoelic injection using a recombinant of acmnpv expressing the enhanced green fluorescent protein gene ... | 2002 | 12075074 |
| distribution and characterization of bacillus thuringiensis on the phylloplane of species of piper (piperaceae) in three altitudinal levels. | bacillus thuringiensis is found naturally on the phylloplane. in this study 35 samples from 13 species of the genus piper (piperaceae) were collected from three altitudinal levels located between 1800 and 2900 m above sea level in the colombian andean forest of central cordillera. two hundred and fifty-six isolates of b. thuringiensis were obtained from 74% of the samples studied. b. thuringiensis index (number of isolates of b. thuringiensis/number of isolates of sporulated bacilli) was 0.2. th ... | 2002 | 12087427 |
| biosynthesis and processing of spodoptera frugiperda alpha-mannosidase iii. | we previously cloned a lepidopteran insect cell cdna that encodes a class ii alpha-mannosidase that is localized in the golgi apparatus but is cobalt-dependent, has a neutral ph optimum, hydrolyzes man(5)glcnac(2) to man(3)glcnac(2), and cannot hydrolyze glcnacman(5)glcnac(2). this enzyme was designated sfmaniii to distinguish it from golgi alpha-mannosidase ii and indicate its derivation from the fall armyworm spodoptera frugiperda. in the present study, we prepared a polyclonal antibody and us ... | 2002 | 12107079 |
| enzyme kinetics and glycan structural characterization of secreted alkaline phosphatase prepared using the baculovirus expression vector system. | secreted human alkaline phosphatase (seap, a model protein containing a single n-glycan chain) was expressed in spodoptera frugiperda sf-9 (sf-9) and trichoplusia ni bti-tn-5b1-4 (tn-5b1-4) insect cell lines infected with recombinant autographa californica multiple nuclear polyhedrovirus expressing seap under control of the polyhedrin promoter. sds-page showed that both systems expressed fairly pure rseap products. the rseap expression level was 7.0 u/ml in tn-5b1-4, higher than the 4.1 u/ml pro ... | 2002 | 12109816 |
| insect antifeedant activity of tetranortriterpenoids from the rutales. a perusal of structural relations. | structure-related insect antifeedant relationship of 56 limonoids (both natural and modified) from the plants belonging to the order rutales was attempted considering substitution patterns, oxidation states, and hydrophobicity, as well as distant geometry derived through conformational analysis on molecular modeling. orientation of the furan and hydroxylation at specific carbon sites have been shown to influence the antifeedancy against the fall armyworm, spodoptera litura. | 2002 | 12137465 |
| combined analysis of supernatant-based feeding bioassays and pcr as a first-tier screening strategy for vip -derived activities in bacillus thuringiensis strains effective against tropical fall armyworm. | to assess whether feeding bioassays using culture-supernatant proteins could be combined with pcr into a first-tier screening strategy for vip3a-like genes efficient against tropical spodoptera frugiperda. | 2002 | 12147075 |
| the role of amino-acid residues q39 and e451 in the determination of substrate specificity of the spodoptera frugiperda beta-glycosidase. | the design of beta-glycosidases with planed substrate specificity for biotechnological application has received little attention. this is mostly a consequence of the lack of data on the molecular basis of the beta-glycosidase specificity, namely data on the energy of the noncovalent interactions in the enzyme-transition state complex. in an attempt to fill this gap, site-directed mutagenesis and enzyme steady-state kinetic experiments with different substrates were conducted, using as model a di ... | 2002 | 12153567 |
| expression of human lactoferrin cdna in insect cells. | a transfer plasmid p8hlfc harboring human lactoferrin cdna was constructed and co-transfected with autographa californica nuclear polyhedrosis virus ( acnpv bacpak6 ) dna into spodoptera frugiperda (sf) cells. the recombinant virus acnpv-hlfc was identified and purified three times by dot hybridization and plaques screening. the expression of human lactoferrin cdna in insect cells was detected by western blotting. the expression level of lactoferrin is about 2% of insect cellular total soluble p ... | 1998 | 12167991 |
| expression, purification, and characterization of human dipeptidyl peptidase iv/cd26 in sf9 insect cells. | the human dipeptidyl peptidase iv/cd26 (dppiv/cd26) is a multifunctional type-ii membrane bound glycoprotein. as a receptor of collagen i and fibronectin it mediates cell-cell and cell-matrix adhesion, and by interacting with extracellular adenosine deaminase and cd45 it is involved in regulatory and costimulatory events in the immune system. dppiv/cd26 has a very distinct substrate specificity, and is potentially capable of truncating many cytokines, chemokines, and peptide hormones. in this st ... | 2002 | 12182835 |
| a approximately 35 kda polypeptide from insect cells binds to yeast acs like elements in the presence of atp. | the s. cerevisiae origin recognition complex binds to the ars consensus sequence in an atp dependent fashion. recently, the yeast cdc6 has been reported to have dna binding activity. conservation of replication proteins among different species strongly supports their functional similarity. here we report the results of an investigation into the dna binding activity of human cdc6 protein. cdc6 was expressed and purified from baculovirus infected sf9 (spodoptera frugiperda) insect cells as gst fus ... | 2002 | 12186657 |
| insect antifeedants from tropical plants ii: structure of zumsin. | a novel a-seco limonoid was isolated from methanolic extract of croton jatrophoides and designated as zumsin. this compound showed potent antifeedant activity against two lepidopteran larvae, pink bollworm, pectinophora gossypiella (pc(50) = 1 microg/cm(2), pc(95) = 8 microg/cm(2)), and fall armyworm, spodoptera frugiperda (pc(50) = 2 microg/cm(2), pc(95) = 16 microg/cm(2)). the structure of zumsin was determined as 1 using a variety of spectroscopic methods including nuclear magnetic resonance, ... | 2002 | 12188606 |
| thiersinines a and b: novel antiinsectan indole diterpenoids from a new fungicolous penicillium species (nrrl 28147). | [structure: see text] two novel antiinsectan indole diterpenoids, thiersinines a (1) and b (2), along with seven known paxilline-type compounds, were isolated from organic extracts of a new penicillium species (p. thiersii nrrl 28147). the structures of 1 and 2 were determined by analysis of 2d nmr data. thiersinines a and b possess a unique spirocyclic subunit that is unprecedented in previously known compounds of this class. both compounds exhibit potent activity against the fall armyworm (spo ... | 2002 | 12201725 |
| a rapid method for measuring intracellular ph using bcecf-am. | a rapid intracellular ph (ph(i)) measurement method based on initial rate of increase of fluorescence ratio of 2',7'-bis(2-carboxyethyl)-5,6-carboxyfluorescein upon dye addition to a cell suspension in growth medium is reported. a dye transport model that describes dye concentration and fluorescence values in intracellular and extracellular spaces provides the mathematical basis for the approach. experimental results of ammonium chloride challenge response of the two suspension cells, spodoptera ... | 2002 | 12204343 |
| heritability and correlation among some selected morphological traits and their relationship with fall armyworm damage in sweet corn. | fall armyworm [spodoptera frugiperda j. e. smith] (faw) is a serious pest in field corn and sweet corn [zea mays l.] in many parts of the world. sweet corn germplasm with effective levels of resistance to damage by the fall armyworm is needed to transfer resistance to commercial hybrids. the objectives of this study were to estimate heritability and to estimate the correlation among some selected morphological traits and their relationship with faw damage. seven shrunken-2 (sh2) inbred lines and ... | 2002 | 12216502 |
| expression of a ligninase by a new hybrid baculovirus with expanded host range. | we have generated and purified a new recombinant baculovirus with expanded host range. acnpv dna and bamhi-digested bmnpv dna were co-transfected into spodoptera frugiperda sf21 cells. progeny viruses were used to infect bmn cells, which are normally resistant to acnpv infection, in order to screen for recombinant viruses with cross-infectivity. a recombinant virus was isolated by plaque purification and analyzed. this isolate was able to infect, replicate and produce polyhedrin in both the sf21 ... | 1997 | 12219209 |
| transport of bile acids in multidrug-resistance-protein 3-overexpressing cells co-transfected with the ileal na+-dependent bile-acid transporter. | many of the transporters involved in the transport of bile acids in the enterohepatic circulation have been characterized. the basolateral bile-acid transporter of ileocytes and cholangiocytes remains an exception. it has been suggested that rat multidrug resistance protein 3 (mrp3) fulfills this function. here we analyse bile-salt transport by human mrp3. membrane vesicles from insect ( spodoptera frugiperda ) cells expressing mrp3 show time-dependent uptake of glycocholate and taurocholate. fu ... | 2003 | 12220224 |
| use of the piggybac transposon for germ-line transformation of insects. | germ-line transformation of insects is now possible with four independent transposable element vector systems. among these, the ttaa-insertion site specific transposon, piggybac, discovered in trichoplusia ni, is one of the most widely used. transformations have been achieved in a wide variety of dipterans, lepidopterans, and a coleopteran, and for many species, piggybac transposition was first tested by plasmid-based mobility assays in cell lines and embryos. all plasmid and genomic insertions ... | 2002 | 12225912 |
| functional expression of a cdna encoding pea (pisum sativum l.) raffinose synthase, partial purification of the enzyme from maturing seeds, and steady-state kinetic analysis of raffinose synthesis. | raffinose (o-alpha- d-galactopyranosyl-(1-->6)- o-alpha- d-glucopyranosyl-(1<-->2)- o-beta- d-fructofuranoside) is a widespread oligosaccharide in plant seeds and other tissues. raffinose synthase (ec 2.4.1.82) is the key enzyme that channels sucrose into the raffinose oligosaccharide pathway. we here report on the isolation of a cdna encoding for raffinose synthase from maturing pea ( pisum sativum l.) seeds. the coding region of the cdna was expressed in spodoptera frugiperda sf21 insect cells ... | 2002 | 12244450 |
| parasitoid-mediated transmission of an iridescent virus. | we examined the interaction between an invertebrate iridescent virus (iiv) isolated from spodoptera frugiperda (j.e. smith) and the solitary ichneumonid endoparasitoid eiphosoma vitticolle cresson. in choice tests, parasitoids examined and stung significantly more virus infected than healthy larvae, apparently due to a lack of defense reaction in virus infected hosts. parasitoid-mediated virus transmission was observed in 100% of the female parasitoids that stung a virus infected host in the lab ... | 2002 | 12384082 |
| p53 autoantibodies in sera from danish ovarian cancer patients and their correlation with clinical data and prognosis. | the p53 gene, a tumour suppressor gene located on the short arm of chromosome 17 (17p13), is frequently mutated in various human tumours. accumulation of p53 protein in neoplastic cells and its release following tumour necrosis can lead to development of circulating autoantibodies (aab) against p53. earlier studies of ovarian cancer (oc) patients reported different frequencies of p53 aab and conclusions regarding the clinical and prognostic value of these aab have not been in agreement. we there ... | 2002 | 12390412 |
| the drosophila diap1 protein is required to prevent accumulation of a continuously generated, processed form of the apical caspase dronc. | although loss of the inhibitor of apoptosis (iap) protein diap1 has been shown to result in caspase activation and spontaneous cell death in drosophila cells and embryos, the point at which diap1 normally functions to inhibit caspase activation is unknown. depletion of the diap1 protein in drosophila s2 cells or the sf-iap protein in spodoptera frugiperda sf21 cells by rna interference (rnai) or cycloheximide treatment resulted in rapid and widespread caspase-dependent apoptosis. co-silencing of ... | 2002 | 12397080 |
| citrus limonoids and their semisynthetic derivatives as antifeedant agents against spodoptera frugiperda larvae. a structure-activity relationship study. | the antifeedant activity of citrus-derived limonoids limonin (1), nomilin (2), and obacunone (3) and their semisynthetic derivatives 4-26 was evaluated against a commercially important pest, spodoptera frugiperda. simple chemical conversions were carried out on the natural limonoids obtained from seeds of citrus limon. these conversions focused on functional groups considered to be important for the biological activity, namely the c-7 carbonyl and the furan ring. in particular, reduction at c-7 ... | 2002 | 12405773 |
| baculovirus lef-12 is not required for viral replication. | the baculovirus lef-12 (orf41) gene is required for transient expression of baculovirus late genes. to analyze the role of lef-12 in the context of infected cells, two mutant viruses were constructed. both mutants were viable in trichoplusia ni high 5 and spodoptera frugiperda sf9 cells. single-step growth curves, however, indicated that virus yields were reduced approximately fivefold in the absence of lef-12. pulse-labeling of infected cells revealed that lef-12 mutant viruses entered the late ... | 2002 | 12414945 |
| production of hybrid double- or triple-layered virus-like particles of group a and c rotaviruses using a baculovirus expression system. | dual infections by group a and group c rotaviruses have been reported, but no reassortants between group a and group c rotaviruses have been described. the vp6 major inner capsid protein of group a and c rotaviruses shares common antigens detected by monoclonal antibodies and also shares 40-43% amino acid identity. coinfection of spodoptera frugiperda (sf9) insect cells with different combinations of the recombinant baculoviruses encoding either group a [rf vp2 (a-vp2), ind vp6 (a-vp6), and vp7 ... | 2002 | 12429511 |
| the response of virally infected insect cells to dissolved oxygen concentration: recombinant protein production and oxidative damage. | the effects of dissolved oxygen (do) concentration on virally infected insect cells were investigated in 3-l bioreactor culture. specifically, cultures of spodoptera frugiperda sf-9 (sf-9) and trichoplusia ni bti-tn-5b1-4 (tn-5b1-4) were infected with autographa californica multiple nucleopolyhedrovirus expressing secreted alkaline phosphatase (seap). following infection at a do concentration of 50% air saturation, the do concentration was adjusted to a final value of either 190%, 50%, or 10% ai ... | 2003 | 12432586 |
| beta-(1 --> 4)-galactosyltransferase activity in native and engineered insect cells measured with time-resolved europium fluorescence. | to evaluate the ability of insect cells to produce complex-type n-glycans, beta-(1 --> 4)-galactosyltransferase (beta4galt) activity in several insect cell lines was analyzed. for this purpose, we developed a simple and highly sensitive assay for beta-(1 --> 4)-galactosyltransferase (beta4galt) activity, which is based on time-resolved fluorometry of europium. bovine serum albumin (bsa) modified with glcnac (glcnac(44)-bsa) was used as the acceptor. glcnac(44)-bsa was coated on a 96-well micropl ... | 2002 | 12433481 |
| functional properties of a rainbow trout cyp3a27 expressed by recombinant baculovirus in insect cells. | cytochrome p450 3a27 (cyp3a27) is highly expressed in liver and intestine of rainbow trout (oncorhynchus mykiss). in many animal species, the intestine and liver are responsible for the first-pass metabolism of a wide range of xenobiotics. to help determine its physiological role, the catalytic capabilities of cyp3a27 protein were examined. an open reading frame of cyp3a27 in pfastbac donor plasmid was transferred to the baculovirus genome (bacmid dna) through tn7 site-specific transposition in ... | 2002 | 12433811 |
| expression of biologically active recombinant equine interferon-gamma by two different baculovirus gene expression systems using insect cells and silkworm larvae. | the full-length equine interferon-gamma (eifn-gamma) cdna, including the secretion signal peptide coding region, was recloned into baculovirus transfer vector pacym1. this vector was co-transfected with autographa californica nuclear polyhedrosis virus dna or hybrid nuclear polyhedrosis virus dna into spodoptera frugiperda cells. the recombinant viruses, named aceifn-gamma and hyeifn-gamma, were then recovered. recombinant eifn-gamma (reifn-gamma) was accumulated in the culture fluid of the acei ... | 2002 | 12445800 |
| neuropeptide co-localisation in the lepidopteran frontal ganglion studied by confocal laser scanning microscopy. | a comparative study of the co-localisation of three different families of neuropeptides, viz. allatostatins of the y/fxfgl-nh(2) type, manduca sexta allatostatin (mas-as) and allatotropin, in the frontal ganglion of lepidopteran larvae has been carried out by means of immunocytochemistry and confocal laser scanning microscopy. the simultaneous application of three types of fluorochrome-conjugated antibodies reveals triple co-localisation in an anterodorsal pair of neurones in the frontal ganglio ... | 2003 | 12483287 |
| effects of temperature on the susceptibility of insect cells to infection by baculoviruses. | three insect cell lines were tested for susceptibility to baculovirus infection by use of a typical endpoint assay procedure. cell lines from spodoptera frugiperda (iplb-sf21ae), lymantria dispar (iplb-ldeita), and heliothis virescens (iplb-hve6s) in 96-well tissue culture plates were each infected with dilutions of extra cellular virus suspensions of the autographa californica nucleopolyhedrovirus (acmnpv). in addition, the l. dispar and h. virescens cells were also infected with l. dispar nucl ... | 2001 | 12486333 |
| monoclonal antibodies to elongation factor-1alpha inhibit in vitro translation in lysates of sf21 cells. | elongation factor-1alpha (ef-1alpha) is an enzyme that is essential for protein synthesis. although ef-1alpha offers an excellent target for the disruption of insect metabolism, agents known to interfere with ef-1alpha activity are toxic to humans. in this article, we describe the development of monoclonal antibodies (mabs) that can disrupt the activity of insect ef-1alpha without cross-reacting with the human enzyme. mabs were generated to ef-1alpha from sf21 cells derived from the fall armywor ... | 2003 | 12489131 |
| endonucleases that may recognize the ssdna-backbone structure: purification from sf9 cells and characterization. | endonucleases that cleave ssdna under conditions that minimize the formation of secondary structures were detected in spodoptera frugiperda sf9 cells. one endonuclease was purified from sf9 cells and another from sf9 cells infected by baculoviruses. polynucleotides containing an a- or t-tract, or a cap binding region were digested in the presence of atp at low mg ions with these two nucleases. atp could be replaced by citrate but not by edta. the cleavage patterns were different from those obtai ... | 2002 | 12489994 |
| functional analysis of a conserved region of the baculovirus envelope fusion protein, ld130. | the envelope fusion protein from a baculovirus pathogenic for lymantria dispar was characterized. n-terminal sequence analysis determined that it was cleaved downstream of predicted signal peptide and furin cleavage motifs. mutation of the furin motif resulted in a protein that was not cleaved and did not mediate fusion. mutagenesis of three charged amino acids in a conserved sequence with the features of a fusion peptide resulted in significant reduction of the ability of the constructs to medi ... | 2002 | 12490405 |
| in vivo induction of apoptosis correlating with reduced infectivity during baculovirus infection. | spodoptera frugiperda caterpillars were infected with a mutant of autographa californica m nucleopolyhedrovirus lacking the antiapoptotic p35 gene. viral infectivity, replication, and spread were substantially reduced compared to that of a control revertant virus. terminal deoxynucleotidyltransferase-mediated dutp-biotin nick end labeling confirmed that apoptosis occurred in mutant-infected caterpillars, thus directly correlating reduced infectivity and in vivo induction of apoptosis. | 2003 | 12525657 |
| n-glycosylation of recombinant human fucosyltransferase iii is required for its in vivo folding in mammalian and insect cells. | human alpha3/4fucosyltransferase (ft3) catalyses the synthesis of fucosylated glycoconjugates involved in cell-cell interactions. ft3 has two potential n-glycosylation sites at asn(154) and asn(185). soluble secretory forms of the enzyme (sft3) and mutant forms with the first, second and both glycosylation sites (sft3dn1, sft3dn2, sft3dn) mutated have been expressed in baby hamster kidney (bhk) and spodoptera frugiperda (sf9) cells. deletion of the first or both sites caused total enzyme inactiv ... | 2003 | 12527108 |
| toxicities of emamectin benzoate homologues and photodegradates to lepidoptera. | the toxicity of a number of emamectin benzoate homologues and photodegradates to five species of lepidoptera was investigated using diet and foliar bioassays. the emamectin benzoate homologues b1a and b1b were equally toxic in the diet and foliar assays to spodoptera exigua (hübner), heliothis virescens (f.), tricoplusia ni (hübner), and spodoptera frugiperda (j. e. smith), within each of these species. plutella xylostella (l.) was the most sensitive species to emamectin benzoate. the ab1a photo ... | 2002 | 12539830 |
| translation of both 5'top and non-top host mrnas continues into the late phase of baculovirus infection. | complete cdna sequences were obtained for ribosomal protein (rp) l15 and eukaryotic initiation factor eif2alpha from the lepidopteran insect spodoptera frugiperda, and for elongation factor eef2 from s. exigua. the presence of a 5' terminal oligopyrimidine (top) tract classified the lepidopteran rpl15 transcript as a top mrna. for eef2, two types of transcripts were observed, one of which had a 5'top tract. the transcript levels for rpl15, eef2 and eif2alpha decreased following baculovirus infec ... | 2003 | 12542638 |
| [isolation, detection toxicity and structure of toxin from beauveria bassiana]. | in this experiment, the crude toxin from the metabolite of beauveria bassiana was isolated and its toxicity was detected. the result shows that indicates toxin has toxicity on larva of aedesalbopictus obvious. though it with low toxicity to larva of heliothis zea when swallowed, but has high toxicity when injected into the blood. for bacteria it only inhibites g+. when it applied to cultured cells of spodoptera frugiperda sf-21, the regression equation of toxicity is y = 2.03 + 2.39x. the half l ... | 1998 | 12548927 |
| sensitivity of invertebrate iridescent virus 6 to organic solvents, detergents, enzymes and temperature treatment. | the sensitivity of invertebrate iridescent virus 6 (iiv-6) to a selection of organic solvents, detergents, enzymes and heat treatment was assayed in spodoptera frugiperda (sf9) cells and by injection of inoculum into larvae of galleria mellonella. in several cases, the degree of sensitivity of the virus depended on the method of assay; cell culture assays indicated greater losses of activity than insect bioassay. iiv-6 was sensitive to chloroform but sensitivity to ether was only detected by cel ... | 2003 | 12573504 |
| improving baculovirus resistance to uv inactivation: increased virulence resulting from expression of a dna repair enzyme. | the use of baculoviruses as biological control agents is hampered by their susceptibility to inactivation by ultraviolet (uv) light. in an attempt to reduce uv inactivation, an algal virus pyrimidine dimer-specific glycosylase, cv-pdg, was expressed in the baculovirus autographa californica m nucleopolyhedrovirus (acmnpv), and the infectivity of recombinant viruses expressing cv-pdg was measured after exposure to uv light. expression of cv-pdg resulted in a 3-fold decrease in inactivation of bud ... | 2003 | 12581719 |