| improvements in spinner-flask designs for insect-cell suspension culture. | experiments have been conducted to characterize the growth of spodoptera frugiperda (fall armyworm) (sf9) cells in standard and modified spinner designs. results suggest that in standard spinner flasks growth is limited by mixing and oxygen transfer. modifications to spinner geometry, baffling and agitator aspect ratio provided increases in growth rate and final cell density. improvements in the performance of spinners, such as those described herein, have the potential to simplify a process and ... | 2003 | 12583814 |
| novel antiinsectan oxalicine alkaloids from two undescribed fungicolous penicillium spp. | 15-deoxyoxalicine b (1) and decaturins a (2) and b (3) have been isolated from penicillium decaturense and penicillium thiersii, two previously undescribed species obtained as colonists of wood-decay fungi. the structures were determined by 2d nmr experiments and/or single-crystal x-ray diffraction analysis. these compounds are members of a rare structural class, and decaturins a and b feature a new polycyclic ring system. decaturin b (3) exhibited potent antiinsectan activity against the fall a ... | 2003 | 12605512 |
| dna microarrays of baculovirus genomes: differential expression of viral genes in two susceptible insect cell lines. | we describe, for the first time, the generation of a viral dna chip for simultaneous expression measurements of nearly all known open reading frames (orfs) in the best-studied members of the family baculoviridae, autographa californica multiple nucleopolyhedrovirus (acmnpv) and bombyx mori nucleopolyhedrovirus (bmnpv). in this study, a viral dna chip (ac-bmnpv chip) was fabricated and used to characterize the viral gene expression profile for acmnpv in different cell types. the viral chip is com ... | 2003 | 12607109 |
| activation of vanilloid receptor type i in the endoplasmic reticulum fails to activate store-operated ca2+ entry. | to evaluate interaction of vanilloid receptor type 1 (trpv1) with endogenous ca(2+) signalling mechanisms, trpv1 was expressed in spodoptera frugiperda (sf 9) insect cells using recombinant baculovirus. stimulation of trpv1-expressing cells, but not control sf 9 cells, with resiniferatoxin (rtx), capsaicin or anandamide, produced an increase in cytosolic free ca(2+) concentration ([ca(2+)](i)), with ec(50) values of 166 pm, 24.5 nm and 3.89 microm respectively. in the absence of extracellular ca ... | 2003 | 12608892 |
| two nonoverlapping domains on the norwalk virus open reading frame 3 (orf3) protein are involved in the formation of the phosphorylated 35k protein and in orf3-capsid protein interactions. | expression of the norwalk virus open reading frame 3 (orf3) in spodoptera frugiperda (sf9) cells yields two major forms, the predicted 23,000-molecular-weight (23k) form and a larger 35k form. the 23k form is able to interact with the orf2 capsid protein and be incorporated into virus-like particles. in this paper, we provide mass spectrometry evidence that both the 23k and 35k forms are composed only of the orf3 protein. two-dimensional gel electrophoresis and phosphatase treatment showed that ... | 2003 | 12610132 |
| maximizing production of estrogen receptor beta with the baculovirus expression system. | steroid hormone/nuclear receptor expression in cultured insect cell lines is routinely driven by a baculovirus vector. an advantage of the baculovirus production of these receptors is that large amounts of functional receptors are obtained for subsequent in vitro studies. most laboratories produce nuclear receptors in spodoptera frugiperda (sf)9 cells. however, no one has determined whether this cell line is optimal for the production of any nuclear receptor. we compared the time course and leve ... | 2003 | 12613256 |
| distribution and diversity of cry genes in native strains of bacillus thuringiensis obtained from different ecosystems from colombia. | colombia is a tropical country located at the north of south america. it is considered to be one of the most important countries in terms of its biodiversity worldwide. one hundred and eight soil samples obtained from agricultural crops and wild ecosystems were evaluated in terms of the presence of bacillus thuringiensis (bt) native strains. one hundred and eight different bt strains were isolated and characterized by the presence of crystal proteins by sds-page and a multiplex pcr with general ... | 2003 | 12623312 |
| cloning, tissue distribution, and functional studies of a new cytochrome p450 3a subfamily member, cyp3a45, from rainbow trout (oncorhynchus mykiss) intestinal ceca. | in trout and mammals, the major extrahepatic expression site for cyp3a forms is in the intestine. a cdna encoding a new cyp3a subfamily member was isolated from rainbow trout intestinal ceca by reverse transcriptase-polymerase chain reaction (rt-pcr), followed by rapid amplification of cdna ends (race)-pcr. in a set of two primers for pcr, a consensus sequence in the highly conserved regions in 17 cyp3a sequences was used for one primer, and the second primer was designed based on adapter sequen ... | 2003 | 12646270 |
| pyroglutamyl-peptidase i: cloning, sequencing, and characterisation of the recombinant human enzyme. | pyroglutamyl-peptidase i (ec 3.4.19.3) is well known from bacteria and archaea, but has not previously been cloned or sequenced from any vertebrate. we describe the cloning and sequencing of the human (aj278828) and mouse (aj278829) forms of pyroglutamyl-peptidase i. the deduced amino acid sequences each consist of 209 residues and show approximately 30% identity with bacterial forms of the enzyme. they show clear homology to the enzyme from prokaryotes and place the mammalian forms of the enzym ... | 2003 | 12651114 |
| 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 | 12658873 |
| human cyp1a1 allelic variants: baculovirus expression and purification, hydrodynamic, spectral, and catalytical properties and their potency in the formation of all-trans-retinoic acid. | three human cytochrome p450 1a1 (cyp1a1) allelic variants, namely wild-type (cyp1a1.1), cyp1a1.2 (i462v), and cyp1a1.4 (t461n), were expressed as c-terminal his-tagged fusions including a thrombin cleavage site in spodoptera frugiperda insect cells by baculovirus infection. the variants were expressed with 30-90 nmol (1.8-5.4 mg) spectrally active cytochrome p450 per one liter of culture and purified to electrophoretic homogeneity by ni-agarose chromatography. the recombinant variants were struc ... | 2003 | 12699690 |
| molecular characterisation of cdnas from the fall armyworm spodoptera frugiperda encoding manduca sexta allatotropin and allatostatin preprohormone peptides. | allatotropin (at) is a 13-residue amidated neuropeptide, first isolated from pharate adult heads of the tobacco hornworm, manduca sexta (manse-at), which strongly stimulates the biosynthesis of juvenile hormones (jh) in the corpora allata (ca) of adult moths. in spodoptera frugiperda, a cdna that encodes 134 amino acids, including an at peptide, has been cloned. the s. frugiperda allatotropin mature peptide (spofr-at) [gfknvemmtargfa] is identical to that isolated from m. sexta. the basic organi ... | 2003 | 12706626 |
| analysis of host specificity of two closely related baculoviruses in permissive and nonpermissive cell lines. | the baculoviruses bombyx mori nucleopolyhedrovirus (bmnpv) and autographa californica multinucleocapsid nucleopolyhedrovirus (acmnpv) share about 90% identity at the genomic level but they have non-overlapping host range and show a high degree of host specificity. we have demonstrated here that acmnpv undergoes dna replication and early gene expression in bombyx-derived bmn cells but fails to show very late gene expression or produce budded virion (bv) particles. coinfection with bmnpv supported ... | 2003 | 12727338 |
| expression and purification of recombinant kaposi's sarcoma-associated herpesvirus dna polymerase using a baculovirus vector system. | the dna polymerase (pol) of kaposi's sarcoma-associated herpesvirus (kshv) is essential for viral dna replication and, thus, may be considered as a viable target for anti-kshv therapeutics. to produce large quantities of homogeneous and pure pol required for high-throughput screening (hts) for inhibitors, we generated a recombinant baculovirus vector encoding a hexahistidine (his6)-tagged pol and infected spodoptera frugiperda sf-9 insect cells. high expression of recombinant pol (rpol) was achi ... | 2003 | 12729724 |
| analysis of two d1-like dopamine receptors from the honey bee apis mellifera reveals agonist-independent activity. | dopamine is found in many invertebrate organisms, including insects, however, the mechanisms through which this amine operates remain unclear. we have expressed two dopamine receptors cloned from honey bee (amdop1 and amdop2) in insect cells (spodoptera frugiperda), and compared their pharmacology directly using production of camp as a functional assay. in each assay, amdop1 receptors required lower concentrations of dopamine and 6,7-adtn for maximal activation than amdop2 receptors. conversely, ... | 2003 | 12750008 |
| a novel medium for expression of proteins selectively labeled with 15n-amino acids in spodoptera frugiperda (sf9) insect cells. | whereas bacterial expression systems are widely used for production of uniformly or selectively (15)n-labeled proteins the usage of the baculovirus expression system for labeling is limited to very few examples in the literature. here we present the complete formulations of the two insect media, iml406 and 455, for the high-yield production of selectively (15)n-labeled proteins in insect cells. the quantities of (15)n-amino acids utilized in the production of labeled gst were similar in the case ... | 2003 | 12766395 |
| expression and in vitro activation of manduca sexta prophenoloxidase-activating proteinase-2 precursor (propap-2) from baculovirus-infected insect cells. | prophenoloxidase activation is a component of the immune system in insects and crustaceans. we recently purified and cloned a new prophenoloxidase-activating proteinase (pap-2) from hemolymph of the tobacco hornworm manduca sexta [j. biol. chem. 278, 3552-3561]. as the terminal component of a putative serine proteinase cascade, this enzyme activates prophenoloxidase (propo) via limited proteolysis. to purify and study the activating proteinase for pap-2 from this insect, we expressed the zymogen ... | 2003 | 12767815 |
| a limited role in parasitism for microplitis demolitor polydnavirus. | spodoptera frugiperda larvae stung by microplitis demolitor undergo physiological alterations characteristic of parasitism. however, despite these physiological modifications, parasitized s. frugiperda larvae never yield adult wasps. our original hypothesis that unsuccessful parasitism was due to a transcriptionally inactive polydnavirus proved untrue. microplitis demolitor polydnavirus (mdpdv) successfully infected and expressed, albeit transiently, in s. frugiperda hemocytes. mdpdv expression ... | 1998 | 12769875 |
| pharmacological characterisation of 5-hydroxytryptamine-induced contractile effects in the isolated gut of the lepidopteran caterpillar spodoptera frugiperda. | the indolealkylamine 5-hydroxytryptamine (5-ht, 0.1nm-1&mgr;m) caused dose-dependent increases in the number of contractions observed in guts isolated from the caterpillar spodoptera frugiperda. of the 5-ht analogues tested for agonist action, 2-methyl-5-ht (0.1-10&mgr;m) was a full agonist with reduced potency while alpha-methyl-5-ht (0.1-100&mgr;m), 5-carboxamidotryptamine (0.1-100&mgr;m), 5-methoxytryptamine (5-meot) (10nm-10&mgr;m), and tryptamine (1-100&mgr;m) were partial agonists. incubat ... | 2002 | 12770131 |
| structure-activity relationship of contractile effects induced by helicokinins in the isolated gut of the lepidopteran caterpillar spodoptera frugiperda. | the diuretic helicokinins yfspwg-amide (hez ki), vrfspwg-amide (hez kii) and kvkfsawg-amide (hez kiii) are potent contractants of the isolated gut of the caterpillar spodoptera frugiperda at doses ranging from 0.1 to 10nm. in comparison, the pentapeptide fspwg-amide was a full agonist with greatly reduced potency while spwg-amide and pwg-amide were weak partial agonists. substitution of individual amino acids in hez ki with alanine revealed that replacement of the [phenylalanine(2)] residue caus ... | 2002 | 12770134 |
| insect larvae contain substances toxic to adults: the discovery of paralysins. | acidic methanolic extracts of larvae of nine different insect species were found to contain substances that cause a lethal effect in the adult stage of the same species and of other species. these endogenous toxic substances, apparently being widely spread over the class of insects, were designated as paralysins, because of their immediate and observable paralytic effect upon injection. the developmental concentration curves of five different species of insects (galleria mellonella (lepidoptera) ... | 1998 | 12770158 |
| proteolytic breakdown of the neb-trypsin modulating oostatic factor (neb-tmof) in the hemolymph of different insects and its gut epithelial transport. | the degradation of the unblocked hexapeptide, trypsin modulating oostatic factor of the flesh fly neobellieria (sarcophaga) bullata (neb-tmof) was studied in vitro in the hemolymph of the lepidopteran spodoptera frugiperda, the orthopteran schistocerca gregaria and the dictyopteran leucophaea maderae. the half-life in the different species varied from approximately 3min in l. maderae to approximately 25min in s. gregaria. purification of the degradation products and esi-qq-oa-tof mass spectromet ... | 2001 | 12770174 |
| ecdysteroid levels/profiles of the parasitoid wasp, diapetimorpha introita, reared on its host, spodoptera frugiperda and on an artificial diet. | diapetimorpha introita is an ichneumonid ectoparasitoid of the fall armyworm, spodoptera frugiperda. since it has been reported that d. introita wasps reared on an artificial diet exhibit a significantly lower percentage of adult eclosion and fecundity than host-reared wasps, this study was undertaken to elucidate the factors responsible for the reduced viability observed in diet-reared wasps. a system of markers has been devised to track the development (from the initiation of cocooning through ... | 2000 | 12770210 |
| nature of the anchors of membrane-bound aminopeptidase, amylase, and trypsin and secretory mechanisms in spodoptera frugiperda (lepidoptera) midgut cells. | spodoptera frugiperda larvae have a microvillar aminopeptidase and both soluble and membrane-bound forms of amylase and trypsin. membrane-bound aminopeptidase is solubilized by glycosyl phosphatidylinositol-specific phospholipase c (gpi-plc) and detergents, suggesting it has a gpi anchor. membrane-bound trypsin is not affected by gpi-plc, although it is solubilized by papain and by different detergents. membrane-bound amylase is similar to trypsin, although once solubilized in detergent it behav ... | 1999 | 12770393 |
| vanessa cardui adipokinetic hormone (vanca-akh) in butterflies and a moth. | small neuropeptides of the adipokinetic hormone/red pigment-concentrating hormone (akh/rpch) family regulate energy metabolism in insects. within lepidopterans, the nonapeptide manduca sexta akh (manse-akh) represents a widely occurring akh, whereas the decapeptide helze-hrth (at first isolated from heliothis zea) seems to be restricted to moths. here we show that vanca-akh, a non-amidated undecapeptide which we recently found in the painted lady butterfly, vanessa cardui, is also present in the ... | 2003 | 12781830 |
| existence of catalase-less peroxisomes in sf21 insect cells. | catalase activity, a peroxisomal marker enzyme, was not detectable in any of the subcellular fractions of spodoptera frugiperda (sf) 21 insect cells, although marker enzymes in other organelles were distributed in the fractions in a manner similar to that seen in mammalian cells. when a green fluorescent protein fused with peroxisome targeting signal 1 at the c-terminal (gfp-skl) was expressed in sf21 cells, punctate fluorescent dots were observed in the cytoplasm. the fraction where gfp-skl was ... | 2003 | 12788084 |
| antioxidants prevent uv-induced apoptosis by inhibiting mitochondrial cytochrome c release and caspase activation in spodoptera frugiperda (sf9) cells. | oxidative stress has been shown to be associated with apoptosis (programmed cell death) in a number of cell systems. we earlier reported in vitro cultured spodoptera frugiperda (sf9) cells as a model system to study oxidative stress induced apoptosis (j biosciences 24 (1999) 13) and the inhibition of uv-induced apoptosis by the baculovirus antiapoptotic p35 protein that acts as a sink to sequester reactive oxygen species (proc natl acad sci usa 96 (1999) 4838). we now show that uv-induced apopto ... | 2003 | 12798776 |
| differential regulation of two arabidopsis type iii phosphatidylinositol 4-kinase isoforms. a regulatory role for the pleckstrin homology domain. | here, we compare the regulation and localization of the arabidopsis type iii phosphatidylinositol (ptdins) 4-kinases, atpi4kalpha1 and atpi4kbeta1, in spodoptera frugiperda (sf9) insect cells. we also explore the role of the pleckstrin homology (ph) domain in regulating atpi4kalpha1. recombinant kinase activity was found to be differentially sensitive to ptdins-4-phosphate (ptdins4p), the product of the reaction. the specific activity of atpi4kalpha1 was inhibited 70% by 0.5 mm ptdins4p. the eff ... | 2003 | 12805633 |
| synthesis of stereospecifically deuterated 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol (nnal) iastereomers and metabolism by a/j mouse lung microsomes and cytochrome p450 2a5. | the tobacco-specific nitrosamine 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (nnk) is a lung carcinogen in mice and rats and is a putative human lung carcinogen. nnk undergoes cytochrome p450-mediated metabolic activation to dna-binding intermediates but is also extensively reduced to 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol (nnal) in vivo. because nnal is also tumorigenic, the carcinogenicity of nnk may actually be governed by the metabolic activation of nnal, rather than direct activat ... | 2003 | 12807362 |
| stereospecific deuterium substitution attenuates the tumorigenicity and metabolism of the tobacco-specific nitrosamine 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (nnk). | stereochemical determinants of the tumorigenicity and metabolism of the tobacco-specific nitrosamine 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (nnk) were investigated using the stereospecifically deuterated isotopomers (4r)-[4-(2)h(1)]nnk and (4s)-[4-(2)h(1)]nnk. upon ip administration to groups of 20 female a/j mice, nnk and (4s)-[4-(2)h(1)]nnk exhibited similar lung tumorigenicity at three different doses, whereas (4r)-[4-(2)h(1)]nnk was 2-fold less tumorigenic at all three doses. in a pa ... | 2003 | 12807363 |
| protein kinase c delta induces src kinase activity via activation of the protein tyrosine phosphatase ptp alpha. | previously we have shown that protein kinase c (pkc)-mediated reorganization of the actin cytoskeleton in smooth muscle cells is transmitted by the non-receptor tyrosine kinase, src. several authors have described how 12-o-tetradecanoylphorbol-13-acetate (tpa) stimulation of cells results in an increase of src activity, but the mechanism of the pkc-mediated src activation is unknown. using pkc isozymes purified from spodoptera frugiperda insect cells, we show here that pkc is not able to activat ... | 2003 | 12826681 |
| functional and biochemical analysis of the type 1 inositol (1,4,5)-trisphosphate receptor calcium sensor. | modulation of the type 1 inositol (1,4,5)-trisphosphate receptors (insp(3)r1) by cytosolic calcium (ca(2+)) plays an essential role in their signaling function, but structural determinants and mechanisms responsible for the insp(3)r1 regulation by ca(2+) are poorly understood. using dt40 cell expression system and ca(2+) imaging assay, in our previous study we identified a critical role of e2100 residue in the insp(3)r1 modulation by ca(2+). by using intrinsic tryptophan fluorescence measurement ... | 2003 | 12829484 |
| purification of dna for the transfection of a spodoptera frugiperda cell line. | spodoptera frugiperda (sf-9) cells have been widely used in baculovirus expression systems, transient gene expression studies and transgenic cell lines. these applications commonly require the transfection of bacterial plasmid dna. one of the most reliable methods of preparing transfection-quality plasmid dna is cesium chloride (cscl) density gradient centrifugation. however, the traditional cscl dna purification is a long and laborious process. we have made a series of modifications to the trad ... | 2002 | 12843704 |
| isolation and characterization of a human sperm antigen gene h-sp-1. | we isolated and characterized a human sperm antigen gene (h-sp-1) from human testis complementary dna using antiserum against the human sperm membrane. northern blot analysis detected two transcripts (2.3 and 1.1 kb) of the h-sp-1 gene. the 2.3-kb transcript is ubiquitous, whereas the 1.1-kb transcript is specific to the human testis with a high level of expression. determination of the base sequence of h-sp-1 showed a size of 2170 bp and 43.4% homology with human synaptophysin. the base sequenc ... | 2003 | 12846798 |
| impact of a nucleopolyhedrovirus bioinsecticide and selected synthetic insecticides on the abundance of insect natural enemies on maize in southern mexico. | the impact of commonly used organophosphate (chlorpyrifos, methamidophos), carbamate (carbaryl), and pyrethroid (cypermethrin) insecticides on insect natural enemies was compared with that of a nucleopolyhedrovirus (baculoviridae) of spodoptera frugiperda (j. e. smith) (lepidoptera noctuidae) in maize grown in southern mexico. analyses of the selectv and koppert side effects (iobc) databases on the impact of synthetic insecticides on arthropod natural enemies were used to predict approximately 7 ... | 2003 | 12852601 |
| field and laboratory evaluations of transgenic cottons expressing one or two bacillus thuringiensis var. kurstaki berliner proteins for management of noctuid (lepidoptera) pests. | field studies were conducted from 1999 to 2001 to evaluate the efficacy of the transgenic cotton, gossypium hirsutum (l.), genotype, bollgard ii (monsanto 15985), which expresses two bacillus thuringiensis berliner (bt) proteins (cry1ac + cry2ab) that are active against lepidopterous pests. bollgard ii was compared with bollgard (dp50b), which expresses only one bt protein (cry1ac), and, in all tests, the conventional variety, dp50, was used as a non-bt control. larval populations of the bollwor ... | 2003 | 12852613 |
| pathogenesis of autographa californica m nucleopolyhedrovirus in fifth instar spodoptera frugiperda. | we have characterized infection and pathogenesis of an autographa californica m nucleopolyhedrovirus recombinant, acmnpv-hsp70/lacz, carrying the lacz reporter gene, in penultimate (fifth) instar spodoptera frugiperda. bioassays revealed that while <0.1 p.f.u. of budded virus was required to generate an ld(50) by intrahaemocoelic injection, approximately 6000 occlusions were required orally to achieve the same mortality in newly moulted fifth instar (5(0)) larvae. in pathogenesis experiments, 78 ... | 2003 | 12867633 |
| baculoviral p35 inhibits oxidant-induced activation of mitochondrial apoptotic pathway. | in this study we report that the baculovirus p35 anti-apoptotic protein prevents cell death by quenching free radicals at a very upstream step in the apoptotic pathway. mitochondria of activated rat peritoneal macrophages as well as spodoptera frugiperda (sf9) insect cells, following treatment with oxidants, h(2)o(2)/uvb irradiation, release cytochrome c followed by activation of caspase-3. transfection of macrophages/sf9 cells with a construct carrying the p35 gene under the cmv/hsp promoters r ... | 2003 | 12893247 |
| transport of fluorescein methotrexate by multidrug resistance-associated protein 3 in iec-6 cells. | the transport characteristics of fluorescein methotrexate (f-mtx) were studied by using the rat intestinal crypt cell line iec-6. enhanced accumulation of f-mtx at 4 degrees c suggests the existence of an active efflux system. mk-571, an inhibitor of the multidrug resistance-associated protein/atp binding cassette c (mrp/abcc) family, also enhanced the accumulation of f-mtx. transcellular transport of f-mtx from the apical to the basolateral compartment was 2.5 times higher than the opposite dir ... | 2003 | 12909565 |
| expression of vp2 gene protein of infectious bursal disease virus detected in korea. | the vp2 gene dna (1.4 kb in approximate) of a very virulent infectious bursal disease virus (vvibdv) chinju strain detected in chinju, korea was cloned into the bacmid, a baculovirus shuttle vector, through transposition of the gene from initially cloned pfastbachta plasmid, a baculovirus expression vector, and was subsequently expressed in spodoptera frugiperda (sf) cells. biological properties of the expressed vp2 subunit protein were characterized to aid in the development of genetically engi ... | 2003 | 12913360 |
| antioxidant and insect growth regulatory activities of stilbenes and extracts from yucca periculosa. | the methanol extract from the bark of yucca periculosa f. baker afforded 4,4'-dihydroxstilbene, resveratrol and 3,3',5,5'-tetrahydroxy-4-methoxystilbene and had growth regulatory activity against the fall army worm (spodoptera frugiperda j.e. smith, lepidoptera:noctuidae) an insect pest of corn. the most active compound was 3,3',5,5'-tetrahydroxy-4-methoxystilbene which had significant effects at 3 microg/g in diets. in addition to the inhibitory activity on bleaching of crocin induced by alkoxy ... | 2003 | 12943764 |
| method to express and purify nm23-h2 protein from baculovirus-infected cells. | high-throughput protein expression and purification are major bottlenecks in the postgenomic and proteomic era. we show here an automated method to express and purify nm23-h2, a nucleoside diphosphate kinase (ndpk), in a 96-well format, by the use of a robotic workstation, from insect spodoptera frugiperda (sf9) baculovirus-infected cells using nickel-nitrilotriacetic acid (ni-nta) agarose beads. the automated method is coupled to mass spectrometry for a validation and quality-control analysis. ... | 2003 | 12951781 |
| characterization of m2 muscarinic receptor activation of different g protein subtypes. | the m2 muscarinic acetylcholine receptor (machr) expressed in insect cells (spodoptera frugiperda, sf9) using the baculovirus system formed active functional complexes with coexpressed gi as well as with go proteins, while no complexes could be detected with internal g proteins. comparison of the abilities of different muscarinic agonists and partial agonists to increase [35s]gtpgammas binding revealed no significant differences between m2/gi and m2/go complexes neither with respect to affinitie ... | 2004 | 12971914 |
| expression of functional anopheles merus alpha-amylase in the baculovirus/spodoptera frugiperda system. | the anopheles merus (diptera, nematocera, culicoidea) alpha-amylase gene (ameramy, genbank accession number u01210) was amplified with its own or with the zabrotes subfasciatusalpha-amylase signal peptide (zsameramy, genbank accession number ay270183) by pcr, using designed primers. the ameramy gene was sequenced from its promotor to the tga codon. as a positive control, the z. subfasciatusalpha-amylase gene with its own signal peptide (zsamy, genbank accession number af255722) was also amplifie ... | 2003 | 12974946 |
| fall armyworm fr sequences map to sex chromosomes and their distribution in the wild indicate limitations in interstrain mating. | the fall armyworm, spodoptera frugiperda, consists of two host strains (rice and corn) that differ in developmental, physiological and behavioural characteristics. however, because the strains are morphologically indistinguishable the investigation of strain-specific behaviour, particularly in the wild, is very difficult. this has spurred the isolation of diagnostic molecular markers. fr sequences are tandem-repeat genetic elements found in large clusters only in the rice strain. to facilitate t ... | 2003 | 12974950 |
| sequence analysis of a 5.1 kbp region of the spodoptera frugiperda multicapsid nucleopolyhedrovirus genome that comprises a functional ecdysteroid udp-glucosyltransferase (egt) gene. | the ecdysteroid udp-glucosyltransferase gene from the spodoptera frugiperda multicapsid nucleopolyhedrovirus (sfmnpv) was identified using degenerate primers whose sequence were derived from conserved regions of the egt proteins encoded by other baculoviruses. analysis of the gene sequence revealed the presence of an open reading frame (orf) with potential to encode a polypeptide of 525 amino acids. promoter sequences typical of baculovirus genes were found in the 5' region of this orf. a polyad ... | 2003 | 14501191 |
| constitutive oligomerization of human d2 dopamine receptors expressed in spodoptera frugiperda 9 (sf9) and in hek293 cells. analysis using co-immunoprecipitation and time-resolved fluorescence resonance energy transfer. | human d2long (d2l) and d2short (d2s) dopamine receptor isoforms were modified at their n-terminus by the addition of a human immunodeficiency virus (hiv) or a flag epitope tag. the receptors were then expressed in spodoptera frugiperda 9 (sf9) cells using the baculovirus system, and their oligomerization was investigated by means of co-immunoprecipitation and time-resolved fluorescence resonance energy transfer (fret). [3h]spiperone labelled d2 receptors in membranes prepared from sf9 cells expr ... | 2003 | 14511374 |
| direct solubilization of heterologously expressed membrane proteins by incorporation into nanoscale lipid bilayers. | one of the biggest challenges in the field of proteomics is obtaining functional membrane proteins solubilized and dispersed into a physiologically relevant environment that maintains the spectrum of in vivo activities. here we describe a system composed of nanoscale self-assembled particles, termed nanodiscs, which contain a single phospholipid bilayer stabilized by an encircling membrane scaffold protein (msp). using microsomal membranes of baculovirus-infected spodoptera frugiperda (sf9) inse ... | 2003 | 14513561 |
| the homologous region sequence (hr1) of autographa californica multinucleocapsid polyhedrosis virus can enhance transcription from non-baculoviral promoters in mammalian cells. | the autographa californica multinucleocapsid polyhedrosis virus homologous region sequence hr1 enhances transcription from the viral polyhedrin promoter in spodoptera frugiperda insect cells and independently functions as an origin of replication (ori) sequence. the binding of the host nuclear protein, hr1-binding protein (hr1-bp), is crucial for the enhancer activity (habib, s., pandey, s., chatterji, u., burma, s., ahmad, r., jain, a., and hasnain, s. e. (1996) dna cell biol. 15, 737-747 and h ... | 2003 | 14570875 |
| evidence for the evolution of ascoviruses from iridoviruses. | ascoviruses (family ascoviridae) are large, enveloped, double-stranded (ds)dna viruses that attack lepidopteran larvae and pupae, and are unusual in that they are transmitted by parasitic wasps during oviposition. previous comparisons of dna polymerase sequences from vertebrate and invertebrate viruses suggested that ascoviruses are closely related to iridoviruses. this relationship was unexpected because these viruses differ markedly in virion symmetry, genome configuration and cellular patholo ... | 2003 | 14573805 |
| autographa californica nucleopolyhedrovirus infection of spodoptera frugiperda cells: a global analysis of host gene regulation during infection, using a differential display approach. | autographa californica nucleopolyhedrovirus (acmnpv), the type member of the virus family baculoviridae, infects pest insects and has been the subject of many studies for its development as a biopesticide. it is also the virus upon which most of the commercial baculovirus protein expression systems are based. acmnpv infection of cultured host spodoptera frugiperda (sf9) cells can induce a number of alterations of host cell properties including altering the cellular cytoskeleton, an arrest of the ... | 2003 | 14573808 |
| rice stripe virus 23.9 k protein aggregates and forms inclusion bodies in cultured insect cells and virus-infected plant cells. | the genome of rice stripe virus (rsv, genus tenuivirus) contains seven open reading frames (orfs). little is known about the products of four of these orfs, including the 23.9 k protein encoded by the virus-sense orf of rna3. western blotting revealed that the 23.9 k protein was synthesized in the host plant and also in the planthopper vector of rsv. using a baculovirus vector, the 23.9 k protein was expressed, both unfused and fused with red-shifted green fluorescent protein, in spodoptera frug ... | 2003 | 14579176 |
| characterization and transcriptional profiles of three spodoptera frugiperda genes encoding cysteine-rich peptides. a new class of defensin-like genes from lepidopteran insects? | the present work describes sequence and transcription of three spodoptera frugiperda genes encoding 6-cysteine-rich peptides. sequence alignments indicate that the predicted peptides belong to the insect defensin family, although phylogenetic analyses suggest they form a cluster distinct from that of other neopteran insect defensins. the three genes were identified in a non-immune-challenged sf9 cells cdna (dna complementary to rna) library (landais et al., bioinformatics, in press) and were nam ... | 2003 | 14597170 |
| defective or effective? mutualistic interactions between virus genotypes. | defective viruses lack genes essential for survival but they can co-infect with complete virus genotypes and use gene products from the complete genotype for their replication and transmission. as such, they are detrimental to the fitness of complete genotypes. here, we describe a mutualistic interaction between genotypes of an insect baculovirus (nucleopolyhedrovirus of spodoptera frugiperda (lepidoptera)) that increases the pathogenicity of the viral population. mixtures of a complete genotype ... | 2003 | 14613611 |
| comparative study on the susceptibility of cutworms (lepidoptera: noctuidae) to agrotis segetum nucleopolyhedrovirus and agrotis ipsilon nucleopolyhedrovirus. | the common cutworm (agrotis segetum) and the black cutworm (agrotis ipsilon) are serious soil pests of many vegetable and field crops all over the world. we have demonstrated the cross-infectivity of two baculoviruses, a. segetum nucleopolyhedrovirus (agsenpv) and a. ipsilon nucleopolyhedrovirus (agipnpv) for these two insect pests. the susceptibility of a. segetum to agipnpv was confirmed by dna restriction endonuclease analyses of dna isolated from virus harvested from infected a. segetum larv ... | 2003 | 14615215 |
| beauvericin cytotoxicity to the invertebrate cell line sf-9. | the cyclic hexadepsipeptide beauvericin, initially known as a secondary metabolite produced by the entomopathogenic fungus beauveria bassiana and toxic to artemia salina larvae, has been more recently recognized as an important mycotoxin synthesized by a number of fusarium strains, which parasite maize, wheat and rice. therefore, this mycotoxin may enter the food chain, causing yet unknown effects to the health of both domestic animals and humans. the cytotoxic effects of beauvericin on mammalia ... | 2003 | 14617831 |
| stable expression of recombinant human alpha3/4 fucosyltransferase iii in spodoptera frugiperda sf9 cells. | human alpha3/4 fucosyltransferase iii (ft3; ec 2.4.1.65) synthesizes fucosylated glycoconjugates, namely the lewis (le) determinants. ft3 is detected in milk, gastric mucosa, kidney and other organs, but is found in very low amounts in these native tissues. in this work, we describe the expression of a soluble secretory form of ft3 (sft3) in spodoptera frugiperda (sf9) insect cells using a non-lytic vector system. the coding sequence was cloned into the expression vector pib/v5-his-topo which co ... | 2003 | 14636711 |
| crystal structure of an invertebrate caspase. | caspases play an essential role in the execution of apoptosis. these cysteine proteases are highly conserved among metazoans and are translated as inactive zymogens, which are activated by proteolytic cleavages to generate the large and small subunits and remove the n-terminal prodomain. the 2.3 a resolution crystal structure of active sf-caspase-1, the principal effector caspase of the insect spodoptera frugiperda, is presented here. the structure represents the first nonhuman caspase to be res ... | 2004 | 14645217 |
| the role of residues r97 and y331 in modulating the ph optimum of an insect beta-glycosidase of family 1. | the activity of the digestive beta-glycosidase from spodoptera frugiperda (sfbetagly50, ph optimum 6.2) depends on e399 (pka = 4.9; catalytic nucleophile) and e187 (pka = 7.5; catalytic proton donor). homology modelling of the sfbetagly50 active site confirms that r97 and y331 form hydrogen bonds with e399. site-directed mutagenesis showed that the substitution of r97 by methionine or lysine increased the e399 pka by 0.6 or 0.8 units, respectively, shifting the ph optima of these mutants to 6.5. ... | 2003 | 14653813 |
| annotation pattern of ests from spodoptera frugiperda sf9 cells and analysis of the ribosomal protein genes reveal insect-specific features and unexpectedly low codon usage bias. | a whole set of expressed sequence tags (ests) from the sf9 cell line of spodoptera frugiperda is presented here for the first time. by this way we want to identify both conserved and specific genes of this pest species. we also expect from this analysis to find a class of protein sequences providing a tool to explore genomic features and phylogeny of lepidoptera. | 2003 | 14668217 |
| (r,s)-reticuline 7-o-methyltransferase and (r,s)-norcoclaurine 6-o-methyltransferase of papaver somniferum - cdna cloning and characterization of methyl transfer enzymes of alkaloid biosynthesis in opium poppy. | s-adenosyl-l-methionine:(r,s)-reticuline 7-o-methyltransferase converts reticuline to laudanine in tetrahydrobenzylisoquinoline biosynthesis in the opium poppy papaver somniferum. this enzyme activity has not yet been detected in plants. a proteomic analysis of p. somniferum latex identified a gel spot that contained a protein(s) whose partial amino acid sequences were homologous to those of plant o-methyltransferases. cdna was amplified from p. somniferum rna by reverse transcription pcr using ... | 2003 | 14675446 |
| synergistic effects of three piper amides on generalist and specialist herbivores. | the tropical rainforest shrub piper cenocladum, which is normally defended against herbivores by a mutualistic ant, contains three amides that have various defensive functions. while the ants are effective primarily against specialist herbivores, we hypothesized that these secondary compounds would be effective against a wider range of insects, thus providing a broad array of defenses against herbivores. we also tested whether a mixture of amides would be more effective against herbivores than i ... | 2003 | 14682530 |
| engineering sialic acid synthetic ability into insect cells: identifying metabolic bottlenecks and devising strategies to overcome them. | previous studies have indicated negligible levels of both sialylation and the precursor n-acetylneuraminic acid (neu5ac) in a number of insect cell lines grown in serum-free medium. the overexpression of the human sialic acid 9-phosphate synthase (sas) in combination with n-acetylmannosamine (mannac) feeding has been shown to overcome this limitation. in this study we evaluated the potential bottlenecks in the sialic acid synthesis pathway in a spodoptera frugiperda (sf9) insect cell line and de ... | 2003 | 14690432 |
| stress-induced apoptosis in spodoptera frugiperda (sf9) cells: baculovirus p35 mitigates eif2 alpha phosphorylation. | spodoptera frugiperda (sf9) ovarian cells, natural hosts for baculovirus, are good model systems to study apoptosis and also heterologous gene expression. we report that uninfected sf9 cells readily undergo apoptosis and show increased phosphorylation of the alpha subunit of eukaryotic initiation factor 2 (eif2alpha) in the presence of agents such as uvb light, etoposide, high concentrations of cycloheximide, and egta. in contrast, tunicamycin, a23187, and low concentrations of cycloheximide pro ... | 2003 | 14690445 |
| evaluation of oils and microbial pathogens for control of lepidopteran pests of sweet corn in new england. | vegetable and mineral oil, beauveria bassiana (balsamo) and bacillus thuringiensis subsp. kurstaki berliner were evaluated for control of ostrinia nubilalis (hübner) (lepidoptera: pyralidae), helicoverpa zea (boddie) (lepidoptera: noctuidae) and spodoptera frugiperda (j. e. smith) (lepidoptera: noctuidae) in sweet corn (zea mays l.). field experiments in maine and massachusetts during 1993 and 1994 evaluated oils and pathogens singly or in combinations, using a single application directly to the ... | 2003 | 14977100 |
| effect of the manganese ion on human alpha3/4 fucosyltransferase iii activity. | the effect of manganese and other divalent cations on the activity of a soluble recombinant form of human alpha3/4 fucosyltransferase iii (sft3) expressed in spodoptera frugiperda (sf9) insect cells was studied. sft3 was active in the absence of divalent cations with an optimum ph of 4.5. in the absence of mn2+ increasing the ph from 4.5 to 7.0 caused a decrease in the affinity of sft3 for the acceptor galbeta3glcnaco(ch2)3nhco(ch2)5nh-biotin, as monitored by the 4-fold increase in the apparent ... | 2004 | 14977360 |
| baculoviruses expressing the human familial alzheimer's disease presenilin 1 mutation lacking exon 9 increase levels of an amyloid beta-like protein in sf9 cells. | presenilin 1 (ps1) plays a pivotal role in the production of the amyloid-beta protein (abeta) that is central to the pathogenesis of alzheimer's disease. ps1 regulates the intramembranous proteolysis of a 99-amino-acid c-terminal fragment of the amyloid precursor protein (app-c99), a cleavage event that releases abeta following a reaction catalyzed by an enzyme termed 'gamma-secretase'. the molecular mechanism of ps1-mediated, gamma-secretase cleavage remains largely unresolved. in particular, c ... | 2004 | 14993906 |
| expression and localization of the spodoptera frugiperda allatotropin (spofr-at) and allatostatin (spofr-as) genes. | manduca sexta allatotropin and allatostatin were the first corpora allata (ca) regulating neuropeptides identified from lepidoptera. recently, we cloned the allatotropin (spofr-at) and the allatostatin (spofr-as) genes from the fall armyworm spodoptera frugiperda. using one-step rt-pcr for semi-quantification of the gene expression, we now demonstrate that three mrna isoforms of the spofr-at gene and the spofr-as gene are expressed in brain, digestive tract, and reproductive organs of larvae, pu ... | 2004 | 15027073 |
| larval and pupal stage of spodoptera frugiperda (j. e. smith) (lepidoptera: noctuidae) in sweet and field corn genotypes. | spodoptera frugiperda, the fall armyworm, is a very significant polyphagous pest due to the damages it causes, and control difficulties. lack of information about its impact on sweet corn motivated a comparison of its biology, with respect to the larval and pupal stages, among the genotypes elisa, br 400 (sweet corns), and br pampa (field corn). in laboratory conditions (25 +/- 1 masculine c; 70 +/- 10% rh; photophase 12 hours), 35 caterpillars were individualized and fed daily with 3.14 cm(2) s ... | 2004 | 15029373 |
| a genomic bac library and a new bac-gfp vector to study the holocentric pest spodoptera frugiperda. | two genomic tools for the study of lepidoptera and the holocentric structure of their chromosomes are presented in this paper. a bacterial artificial chromosome (bac) library was constructed using nuclear dna partially digested with hindiii from eggs of spodoptera frugiperda. the library contains a total of 36,864 clones with an average insert size of 125 kb, which corresponds to approximately 11.5 genome equivalents. hybridization screening of the library was performed with eight single-copy ge ... | 2004 | 15041017 |
| arachidonic and eicosapentaenoic acid metabolism by human cyp1a1: highly stereoselective formation of 17(r),18(s)-epoxyeicosatetraenoic acid. | human cytochrome p450 1a1 (cyp1a1) and human nadph-cytochrome p450 reductase were expressed and purified from spodoptera frugiperda insect cells. a reconstituted enzymatically active system metabolized polyunsaturated fatty acids such as arachidonic (aa) and eicosapentaenoic acid (epa). cyp1a1 was an aa hydroxylase which oxidizes this substrate at a rate of 650+/-10 pmol/min/nmol cyp1a1, with over 90% of metabolites accounted for by hydroxylation products and with 19-oh-aa as major product. epox ... | 2004 | 15041462 |
| cytotoxicity of fungal metabolites to lepidopteran (spodoptera frugiperda) cell line (sf-9). | the toxicity of sixteen fungal metabolites produced by some entomopathogenic fungi or biological control fungi agents was evaluated on lepidopteran spodoptera frugiperda (sf-9) cell line by trypan blue dye exclusion and mtt-colorimetric assay, after 48 h of incubation. no statistical difference was found between ic50values (50% inhibiting concentration) and cc50 values (50% cytotoxicity concentration) obtained by mtt test and trypan blue dye exclusion for each fungal metabolite. by mtt assay, th ... | 2004 | 15050836 |
| in vivo binding of the cry11bb toxin of bacillus thuringiensis subsp. medellin to the midgut of mosquito larvae (diptera: culicidae). | bacillus thuringiensis subsp. medellin produces numerous proteins among which 94 kda known as cry11bb, has mosquitocidal activity. the mode of action of the cry11 proteins has been described as similar to those of the cry1 toxins, nevertheless, the mechanism of action is still not clear. in this study we investigated the in vivo binding of the cry11bb toxin to the midgut of the insect species anopheles albimanus, aedes aegypti, and culex quinquefasciatus by immunohistochemical analysis. spodopte ... | 2004 | 15057351 |
| rapid quantification of juvenile hormones and their metabolites in insect haemolymph by liquid chromatography-mass spectrometry (lc-ms). | a simple, fast and sensitive method was developed for routine determination of juvenile hormone (jh), jh diols and jh acids in insect haemolymph, by liquid chromatography-mass spectrometry (lc-ms). sample clean-up involves the precipitation of proteins by methanol/isooctane (1:1, v/v), centrifugation and partial evaporation of the organic solvents. since jh is bound to a carrier protein in the haemolymph, a binding protein (bp) assay was performed to ensure jh is removed during precipitation. th ... | 2004 | 15057500 |
| conservation of a proteinase cleavage site between an insect retrovirus (gypsy) env protein and a baculovirus envelope fusion protein. | the predicted env protein of insect retroviruses (errantiviruses) is related to the envelope fusion protein of a major division of the baculoviridae. the highest degree of homology is found in a region that contains a furin cleavage site in the baculovirus proteins and an adjacent sequence that has the properties of a fusion peptide. in this investigation, the homologous region in the env protein of the gypsy retrovirus of drosophila melanogaster (dmegypv) was investigated. alteration of the pre ... | 2004 | 15063117 |
| silencing structural and nonstructural genes in baculovirus by rna interference. | we review several aspects of rnai and gene silencing with baculovirus. we show that the potency of rnai in spodoptera frugiperda (sf21) insect cells correlates well with the efficiency of transfection of the sirna. using a fluorescein-labeled sirna we found that the sirna localized in areas surrounding the endoplasmic reticulum (er). both long (700 nucleotides long) and small ( approximately 25 nucleotides long) interfering rnas were equally effective in initiating rna interference (rnai), and t ... | 2004 | 15068883 |
| purification and characterization of recombinant human prostacyclin synthase. | prostacyclin synthase (pgis), which catalyzes the conversion of prostaglandin (pg) h(2) to prostacyclin (pgi(2)), is a member of the cytochrome p-450 (p450) superfamily, cyp8a1. to study the enzymatic and protein characteristics of human pgis, the enzyme was overexpressed in spodoptera frugiperda 21 (sf21) cells using the baculovirus expression system. pgis was expressed in the microsomes of the infected sf21 cells after culture in 5 microg/ml hematin-supplemented medium for 72 h. the holoenzyme ... | 2004 | 15115769 |
| effects of elevated atmospheric co2 on the nutritional ecology of c3 and c4 grass-feeding caterpillars. | it is plausible that the nutritional quality of c3 plants will decline more under elevated atmospheric co2 than will the nutritional quality of c4 plants, causing herbivorous insects to increase their feeding on c3 plants relative to c4 plants. we tested this hypothesis with a c3 and c4 grass and two caterpillar species with different diet breadths. lolium multiflorum (c3) and bouteloua curtipendula (c4) were grown in outdoor open top chambers at ambient (370 ppm) or elevated (740 ppm) co2. bioa ... | 2004 | 15118901 |
| structure and function of recombinant cobra venom factor. | cobra venom factor (cvf) is the complement-activating protein from cobra venom. it is a structural and functional analog of complement component c3. cvf functionally resembles c3b, the activated form of c3. like c3b, cvf binds factor b, which is subsequently cleaved by factor d to form the bimolecular complex cvf,bb. cvf,bb is a c3/c5 convertase that cleaves both complement components c3 and c5. cvf is a three-chain protein that structurally resembles the c3b degradation product c3c, which is un ... | 2004 | 15131128 |
| a constitutive super-enhancer: homologous region 3 of bombyx mori nucleopolyhedrovirus. | transient expression assays were carried out to assess the transcriptional enhancement from the immediate-early gene (ie-1) promoters by cis-linked bombyx mori nucleopolyhedrovirus (bmnpv) homologous region-3 (hr3). the ie-1 promoters were derived from autographa californica multinucleocapsid nucleopolyhedrovirus (acmnpv) and bmnpv. hr3 was placed downstream of the luciferase (luc) gene, the resulting plasmids were used to transfect the spodoptera frugiperda (sf-21), bombyx mori (bm-5, bm-n) cel ... | 2004 | 15147978 |
| assessment of experimental bt events against fall armyworm and corn earworm in field corn. | performance of experimental bacillus thuringiensis (bt) mon events alone and pyramided with mon810 were evaluated over 3 yr in georgia and alabama. ability of events to prevent whorl defoliation by the fall armyworm, spodoptera frugiperda (j. e. smith), and natural ear feeding damage by the corn earworm, helicoverpa zea (boddie) was assessed. in each year, near-isogenic hybrids with novel single transformation events and crosses pyramided with the mon810 event were compared with the standard sin ... | 2004 | 15154443 |
| ultralow rates of spinosad in phagostimulant granules provide control of spodoptera frugiperda (lepidoptera: noctuidae) in maize. | field trails in 2002 and 2003 were performed to determine the efficacy of maize flour-based granular formulations with ultralow rates of the naturally derived insecticide spinosad (0.1, 0.3, and 1.0 g [ai]/ha), for control of spodoptera frugiperda (j.e. smith) in maize, zea mays l., in southern mexico. spinosad formulations were compared with a chemical standard, a commercial granular formulation of chlorpyrifos (150 g [ai]/ha). in both years, application of spinosad resulted in excellent levels ... | 2004 | 15154464 |
| comparison of anti-feedant and insecticidal activity of nimbin and salannin photo-oxidation products with neem (azadirachta indica) limonoids. | photo-oxidation of the neem limonoids nimbin and salannin with uv light in the presence of oxygen gives two isomeric lactone products per limonoid, nimbinolide and isonimbinolide, and salanninolide and isosalanninolide, respectively. when compared in insect tests with the important limonoids of neem seeds, azadirachtin, nimbin and salannin, isonimbinolide and isosalanninolide show activity greater than that of nimbin or salannin and in some respects show activity approaching that of azadirachtin ... | 2004 | 15154512 |
| biological activity of phenolic compounds from alchornea glandulosa. | from the isopropyl acetate fraction obtained by partition of the leaves ethanolic extract of alchornea glandulosa, six phenolic compounds were isolated and identified. general lethality of the extract, fractions and compounds were assayed with brine shrimp test (bst) and with antifeedant activity bioassay (aab) against neonate larvae of spodoptera frugiperda. aqueous fraction and ethyl gallate, one of the major constituents of the polar fraction, have shown the highest toxicity in bst. ethanolic ... | 2004 | 15159004 |
| recombinant cobra venom factor. | cobra venom factor (cvf) is the complement-activating protein from cobra venom. cvf is a three-chain protein that functionally resembles c3b, the activated form of complement component c3. like c3b, cvf forms a c3/c5 convertase with factor b in the presence of factor d and mg(2+). although cvf exhibits functional activity of c3b, it structurally resembles the c3b degradation product c3c, which is not able to form a c3/c5 convertase. cvf has become an important research tool to decomplement labor ... | 2004 | 15159065 |
| insect antifeedants from tropical plants: structures of dumnin and dumsenin. | two novel a-seco limonoids, dumnin and dumsenin, were isolated from the methanolic extract of croton jatrophoides by bioassay-guided fractionation, and the structures were determined by nuclear magnetic resonance, circular dichroism, and mass spectrometry experiments. these compounds showed potent antifeedant activity (pc(50) <or= 2.0 microg/ml) against the larvae of pink bollworm, pectinophora gossypiella, and/or fall armyworm, spodoptera frugiperda, providing results comparable to dumsin and z ... | 2004 | 15161191 |
| the rhopalosiphum padi virus 5' internal ribosome entry site is functional in spodoptera frugiperda 21 cells and in their cell-free lysates: implications for the baculovirus expression system. | cap-independent internal initiation of translation occurs on a number of viral and cellular mrnas and is directed by internal ribosome entry site (ires) elements. rhopalosiphum padi virus (rhpv) is a member of the dicistroviridae. these viruses have single-stranded, positive-sense rna genomes that contain two open reading frames, both preceded by ires elements. previously, the activity of the rhpv 5' utr ires has been demonstrated in mammalian, drosophila and wheat germ in vitro translation syst ... | 2004 | 15166440 |
| cloning and expression of human udp-glucuronosyltransferase 1a4 in bac-to-bac system. | udp-glucuronosyltransferases (ugts) catalyze the transfer of glucuronic acid from uridine diphosphate-glucuronic acid (udp-ga) to compounds with amine, hydroxyl, and carboxylic acid moieties. n-glucuronidation is an important pathway for elimination of many tertiary amine therapeutic agents used in humans. ugt1a4 has been reported to be specific for glucuronidating primary, secondary, and tertiary amines, forming n-glucuronides. to further investigate the drugs metabolized by ugt1a4, the bac-to- ... | 2004 | 15178418 |
| inhibition of insulin-like growth factor i receptor autophosphorylation by novel 6-5 ring-fused compounds. | the insulin-like growth factor 1 receptor (igf1r) plays an important role in cell transformation, and it has emerged as a target for anti-cancer drug design. igf1r is activated by autophosphorylation at three sites in the enzyme activation loop. we describe here a group of 6-5 ring-fused compounds that are the first reported inhibitors selective for the unphosphorylated (0p) form of igf1r. these compounds do not significantly inhibit the fully activated, triply phosphorylated (3p) form. igf1r wa ... | 2004 | 15183126 |
| radiation responses of sf9, a highly radioresistant lepidopteran insect cell line. | lepidopteran insect cells are known to exhibit very high radioresistance. although very effective dna excision-repair has been proposed as a contributing factor, a detailed understanding of insect cell radiation responses has not yet been obtained. therefore, the study was carried out to understand the in vitro radiation responses of sf9 lepidopteran cells. | 2004 | 15204707 |
| large-scale overproduction, functional purification and ligand affinities of the his-tagged human histamine h1 receptor. | this report describes an efficient strategy for amplified functional purification of the human h1 receptor after heterologous expression in sf9 cells. the cdna encoding a c-terminally histidine-tagged (10xhis) human histamine h1 receptor was used to generate recombinant baculovirus in a spodoptera frugiperda-derived cell line (iplb-sf9). as judged from its ligand affinity profile, functional receptor could be expressed at high levels (30-40 pmol per 10(6) cells). rapid proteolysis in the cell cu ... | 2004 | 15206929 |
| type-a allatostatins from the fall armyworm, spodoptera frugiperda: molecular cloning, expression and tissue-specific localization. | the gene encoding the spodoptera frugiperda allatostatin type-a peptide family (y/fxfgl-amides) was isolated from s. frugiperda brain cdna. the gene encodes a precursor of 231 amino acids containing nine (or ten) y/fxfgl-a peptides that are tandemly arranged in three blocks. the comparison of the spofr-ast a precursor with the respective precursor genes from two other lepidopteran species, helicoverpa armigera and bombyx mori, shows high homology in size, sequence (84 and 57%, respectively), and ... | 2004 | 15211550 |
| sirna-directed silencing of transgene expressed in cultured insect cells. | rna interference (rnai) has emerged as a powerful tool to rapidly analyze gene functions in a wide variety of eukaryotic organisms as well as in cultured cell lines. we demonstrate here that rnai can be applied to study the function of a transgene expressed in an insect cell line (spodoptera frugiperda, sf21). the aminopeptidase n gene (apn) targeted for silencing in the present study was isolated from the midgut of spodoptera litura larvae and expressed in sf21 cells using baculovirus expressio ... | 2004 | 15219846 |
| characterization of a lipid activated ctp:phosphocholine cytidylyltransferase from drosophila melanogaster. | ctp:phosphocholine cytidylyltransferase (cct) is an enzyme critical for cellular phosphatidylcholine (pc) synthesis, converting phosphocholine and cytidine 5'-triphosphate (ctp) to cdp-choline. we have isolated a cdna encoding an isoform of cct from drosophila melanogaster and expressed the recombinant native and 6 x -his-tagged forms using a baculovirus expression system in spodoptera frugiperda (sf9) insect cells. immunoblot using anti-phospho amino acid antibodies reveals the enzyme is phosph ... | 2004 | 15238222 |
| purified recombinant human prosaposin forms oligomers that bind procathepsin d and affect its autoactivation. | before delivery to endosomes, portions of procd (procathepsin d) and prosap (prosaposin) are assembled into complexes. we demonstrate that such complexes are also present in secretions of cultured cells. to study the formation and properties of the complexes, we purified procd and prosap from culture media of spodoptera frugiperda cells that were infected with baculoviruses bearing the respective cdnas. the biological activity of procd was demonstrated by its ph-dependent autoactivation to pseud ... | 2004 | 15255780 |
| characterization of a baculovirus with a deletion of vlf-1. | very late expression factor (vlf-1) of autographa californica multiple nucleopolyhedrovirus (acmnpv) is essential for high levels of expression of the very late genes p10 and polh, and evidence suggests vlf-1 may also be involved in viral dna replication. in this study, investigations determined whether vlf-1 is essential for viral dna replication by generating a vlf-1 knockout bacmid containing the acmnpv genome through homologous recombination in escherichia coli. additionally, a vlf-1 repair ... | 2004 | 15262507 |
| lysosomal acid lipase as a preproprotein. | lysosomal acid lipase (lal; ec 3.1.1.13) hydrolyzes intracellular triglycerides and cholesterol esters taken up by various cell-types. previously, lal purified from human liver tissue was described as a preproprotein with a 27 amino acid signal peptide and a 49 amino acid propeptide. three mutants of the putative proregion of lal were produced and expressed in spodoptera frugiperda insect cells. pulse-chase experiments demonstrated that lal undergoes proteolytical processing. the deletion of the ... | 2004 | 15269241 |
| insect growth inhibition by tocotrienols and hydroquinones from roldana barba-johannis. | the methanol extract from the aerial parts of roldana barba-johannis (asteraceae) afforded sargachromenol, sargahydroquinoic acid, and sargaquinoic acid. these natural products and their corresponding acetylated and methylated derivatives showed insecticidal and insect growth regulatory activities against the fall armyworm [spodoptera frugiperda j.e. smith, (lepidoptera: noctuidae)], an important insect pest of corn. the most active compounds were sargachromenol and its acetylated derivative; sa ... | 2004 | 15280003 |
| differentially enhanced insect resistance, at a cost, in arabidopsis thaliana constitutively expressing a transcription factor of defensive metabolites. | a transgenic line of arabidopsis thaliana constitutively expressing a conserved myb transcription factor of phenylpropanoid biosynthesis resulting in solid-purple leaves had significantly increased resistance to leaf feeding by first instar fall armyworms (spodoptera frugiperda), but no enhanced resistance to cabbage looper (trichoplusia ni) larvae, when compared to wild type plants. however, inflorescence and silique (seed pod) production were significantly reduced by 22 and 52%, respectively, ... | 2004 | 15291486 |
| competition of bacillus thuringiensis cry1 toxins for midgut binding sites: a basis for the development and management of transgenic tropical maize resistant to several stemborers. | binding and competition of five bacillus thuringiensis toxins--cry1ab, cry1ac, cry1ba, cry1ca, and cry1ea--for midgut binding sites from three pests, spodoptera frugiperda, diatraea saccharalis, and diatraea grandiosella, were investigated as part of a strategy to develop tropical transgenic maize resistant to several stemborers. on s. frugiperda, cry1ab and cry1ac compete for the same binding site; cry1ba and cry1ca compete for a second binding site. cry1ea recognizes a third specific binding s ... | 2004 | 15297925 |