| insect cells as hosts for the expression of recombinant glycoproteins. | baculovirus-mediated expression in insect cells has become well-established for the production of recombinant glycoproteins. its frequent use arises from the relative ease and speed with which a heterologous protein can be expressed on the laboratory scale and the high chance of obtaining a biologically active protein. in addition to spodoptera frugiperda sf9 cells, which are probably the most widely used insect cell line, other mainly lepidopteran cell lines are exploited for protein expression ... | 1999 | 10612411 |
| biological activities of crude plant extracts from vitex trifolia l. (verbenaceae). | biological assays of vitex trifolia l. organic extracts have shown relevant activities. hexanic and dichloromethanic (dcm) extracts, when prepared from stems and foliage, have proved to be very toxic against several cancer cell lines in culture (sqc-1 uiso, ovcar-5, hct-15 coladcar, and kb). also, an important antifeeding activity against the insect pest spodoptera frugiperda (lepidoptera: noctuidae) was recorded. the hexanic extract from leaves completely inhibited the growth of the fungal plan ... | 1999 | 10616958 |
| expression of fas-fas ligand in murine testis. | during spermatogenesis, it has been suggested that the number of germ cells to be matured is regulated and restricted through the apoptotic mechanism. in the present study, we investigated the expression and apoptotic role of fas and fas ligand (l) in the murine testis. | 1999 | 10622469 |
| analysis of tissue inhibitor of metalloproteinases-2 effect on pro-matrix metalloproteinase-2 activation by membrane-type 1 matrix metalloproteinase using baculovirus/insect-cell expression system. | membrane-type 1 matrix metalloproteinase (mt1-mmp; mmp14) is known to activate pro-matrix metalloproteinase-2 (pro-mmp-2; progelatinase a) on the cell surface. to analyse the tissue inhibitor of metalloproteinases-2 (timp-2) effect on activation of pro-mmp-2 by mt1-mmp, we have expressed the full-size mt1-mmp (fmt1-mmp) and a transmembrane (tm)-domain-deleted soluble mt1-mmp (smt1-mmp) in the baculovirus/sf9 (spodoptera frugiperda 9) insect-cell system, where neither endogenous gelatinolytic mmp ... | 2000 | 10642509 |
| use of flow cytometry to rapidly optimize the transfection of animal cells. | plasmid transfection is the first step in the generation of stably transformed animal cells and is also a useful tool for analyzing transient gene expression. maximizing the transfection efficiency and expression level from the introduced plasmid is critical to the success of these processes. by means of lipid-mediated transfection, a plasmid vector expressing the green fluorescence reporter protein has been coupled with flow cytometry to conveniently investigate those parameters that impact the ... | 2000 | 10649786 |
| stachyose synthesis in seeds of adzuki bean (vigna angularis): molecular cloning and functional expression of stachyose synthase. | stachyose is the major soluble carbohydrate in seeds of a number of important crop species. it is synthesized from raffinose and galactinol by the action of stachyose synthase (ec 2.4.1.67). we report here on the identification of a cdna encoding stachyose synthase from seeds of adzuki bean (vigna angularis ohwi et ohashi). based on internal amino acid sequences of the enzyme purified from adzuki bean, oligonucleotides were designed and used to amplify corresponding sequences from adzuki bean cd ... | 1999 | 10652123 |
| hybrid and complex glycans are linked to the conserved n-glycosylation site of the third eight-cysteine domain of ltbp-1 in insect cells. | covalent association of ltbp-1 (latent tgf-beta binding protein-1) to latent tgf-beta is mediated by the third eight-cysteine (also referred to as tb) module of ltbp-1, a domain designated as cr3. spodoptera frugiperda (sf9) cells have proved a suitable cell system in which to study this association and to produce recombinant cr3, and we show here that another lepidopteran cell line, trichoplusia nitn-5b1-4 (high-five) cells, allows the recovery of large amounts of functional recombinant cr3. cr ... | 2000 | 10677208 |
| evolutionary conservation of apoptosis mechanisms: lepidopteran and baculoviral inhibitor of apoptosis proteins are inhibitors of mammalian caspase-9. | we cloned a new inhibitor of apoptosis protein (iap) homolog, sfiap, from spodoptera frugiperda sf-21 cells, a host of insect baculoviruses. sfiap contains two baculovirus iap repeat domains followed by a ring domain. sfiap has striking amino acid sequence similarity with baculoviral iaps, cpiap and opiap, suggesting that baculoviral iaps may be host-derived genes. sfiap and baculoviral cpiap inhibit bax but not fas-induced apoptosis in human cells. their apoptosis-suppressing activity in mammal ... | 2000 | 10677478 |
| expression of a neuronal nicotinic acetylcholine receptor in insect and mammalian host cell systems. | different mammalian and insect somatic host cell systems were tested in their ability to express, fold, and assemble alpha7-type neuronal acetylcholine receptor (achr) both at the transcriptional and translational level. for this purpose we employed clonal cell lines derived from the neural crest, such as pc12 cells from a rat adrenal pheochromocytoma, and gh3 cells isolated from a rat pituitary tumor, as well as non-neuronal cells such as nih-3t3 fibroblasts from embryonic nih swiss mouse and s ... | 2000 | 10685617 |
| induction of apoptosis in glioma cells by recombinant human fas ligand. | fas ligand (fasl) belongs to the tumor necrosis factor family and has the ability to induce apoptosis in susceptible target cells by binding to its receptor, fas. it has been demonstrated recently that the fasl/fas system plays a pivotal role in the cytocidal activity of t lymphocytes in the immune system. fasl may act as a cytotoxic effector molecule to fas-expressing malignant tumor cells. we reported previously that fas is commonly expressed in human brain tumor cells. in this study, we exami ... | 2000 | 10690733 |
| kinetics studies of the cardiac ca-atpase expressed in sf21 cells: new insights on ca-atpase regulation by phospholamban. | kinetics studies of the cardiac ca-atpase expressed in sf21 cells (spodoptera frugiperda insect cells) have been carried out to test the hypotheses that phospholamban inhibits ca-atpase cycling by decreasing the rate of the e1.ca to e1'.ca transition and/or the rate of phosphoenzyme hydrolysis. three sample types were studied: ca-atpase expressed alone, ca-atpase coexpressed with wild-type phospholamban (the natural pentameric inhibitor), and ca-atpase coexpressed with the l37a-phospholamban mut ... | 2000 | 10692318 |
| a single glycine residue at the entrance to the first membrane-spanning domain of the gamma-aminobutyric acid type a receptor beta(2) subunit affects allosteric sensitivity to gaba and anesthetics. | site-directed mutagenesis of the gamma-aminobutyric acid type a (gaba(a)) receptor beta(2) subunit has demonstrated that conversion of a conserved glycine residue located at the entrance to the first transmembrane domain into the homologous rho(1) residue phenylalanine alters the modulating effects of four different i.v. anesthetics: pentobarbital, alphaxalone, etomidate, and propofol. using the baculovirus expression system in spodoptera frugiperda 9 cells, anesthetic-induced enhancement of [(3 ... | 2000 | 10692487 |
| pathways of glutamine metabolism in spodoptera frugiperda (sf9) insect cells: evidence for the presence of the nitrogen assimilation system, and a metabolic switch by 1h/15n nmr. | 1h/15n and 13c nmr were used to investigate metabolism in spodoptera frugiperda (sf9) cells. labelled substrates ([2-15n]glutamine, [5-15n]glutamine, [2-15n]glutamate, 15nh4cl, [2-15n]alanine, and [1-13c]glucose) were added to batch cultures and the concentration of labelled excreted metabolites (alanine, nh4+, glutamine, glycerol, and lactate) were quantified. cultures with excess glucose and glutamine produce alanine as the main metabolic by-product while no ammonium ions are released. 1h/15n ... | 2000 | 10702908 |
| cloning, sequencing and expression of the gene that encodes the major neutralisation-specific antigen of african horsesickness virus serotype 9. | a marked improvement in the efficiency of cloning the large double stranded rna (dsrna) genome segments of african horsesickness virus (ahsv) was achieved when the dsrna polyadenylation step was carried out with undenatured rather than strand-separated dsrna. it is a prerequisite to use dsrna of very high purity because in the presence of even trace amounts of single stranded rna, the dsrna appears to be poorly polyadenylated as judged by its effectiveness as a template for oligo-dt-primed cdna ... | 2000 | 10713375 |
| adaptation of an insect cell line of spodoptera frugiperda to grow at 37 degrees c: characterization of an endodiploid clone. | sf21 and sf9 cell lines established from the lepidoptera spodoptera frugiperda do not display major induction of heat shock proteins when exposed to a temperature of 37 degrees c. after some months of adaptation at 37 degrees c we obtained two new cell lines, sf21-ht and sf9-ht, which have now been established for several years in our laboratory. the sf9-ht line displays a slightly shorter doubling time at 37 degrees c than the wild type at 28 degrees c, but cell lethality gives rise to an earli ... | 2000 | 10718368 |
| interactions of the human multidrug resistance proteins mrp1 and mrp2 with organic anions. | the human multidrug resistance protein mrp1 and its homolog, mrp2, are both suggested as being involved in cancer drug resistance and the transport of organic anions. we expressed mrp1 and mrp2 in spodoptera frugiperda ovarian cells and compared their atp-dependent transport properties and vanadate-sensitive atpase activities in isolated membrane vesicles. both mrp1 and mrp2 actively transported leukotriene c(4) and n-ethylmaleimide glutathione (nem-gs), although the relative affinity of mrp2 fo ... | 2000 | 10727523 |
| the multifunctional deoxynucleoside kinase of insect cells is a target for the development of new insecticides. | the antiherpetic agent (e)-5-(2-bromovinyl)-2'-deoxyuridine (bvdu) was found to be an efficient substrate for recombinant drosophila melanogaster-deoxyribonucleoside kinase with a k(m) of 4.5 microm and a v(max) of 400 nmol/microg protein/h compared with 1.3 microm and 62.5 nmol/microg protein/h, respectively, for the natural substrate thymidine. mammalian cytosolic thymidine kinase-1 does not recognize bvdu as a substrate. in sharp contrast to mammalian cells, the insect d. melanogaster and spo ... | 2000 | 10727530 |
| synthesis and use of 3'-(azidoiodosalicyl) derivatives of 2', 5'-dideoxyadenosine as photoaffinity ligands for adenylyl cyclase. | 3'-[(4-azidosalicyl)glycyl]-2',5'-dideoxyadenosine (1), 3'- [(4-azidosalicyl)-gamma-aminobutyryl]-2',5'-dideoxyadenosine (2), and the (125)i-labeled mono- and diiodinated analogs of 1 were synthesized and tested as photoaffinity probes for adenylyl cyclases. kinetics for inhibition of purified type i enzyme by 1 was noncompetitive with respect to mn(*)5'-atp in the absence of light, implying a p-site mechanism of inhibition. in a uv-dependent manner both 1 and 2 and the iodinated derivative of 1 ... | 2000 | 10729209 |
| hematopoiesis in larval pseudoplusia includens and spodoptera frugiperda. | maintenance of circulating hemocytes in larval lepidoptera has been attributed to both mitosis of hemocytes already in circulation and the release of hemocytes from hematopoietic organs. in this study, we compared hematopoiesis in the noctuids pseudoplusia includens and spodoptera frugiperda. for both species, hemocyte densities per microl of blood increased with instar. differential hemocyte counts indicated that plasmatocytes were the most abundant hemocyte type during early instars but granul ... | 2000 | 10737919 |
| a new secreted insect protein belonging to the immunoglobulin superfamily binds insulin and related peptides and inhibits their activities. | insulin and related peptides are key hormones for the regulation of growth and metabolism. here we describe a novel high affinity insulin-related peptide-binding protein (ibp) secreted from cells of the insect spodoptera frugiperda. this ibp is composed of two ig-like c2 domains, has a molecular mass of 27 kda, binds human insulin with an affinity of 70 pm, and inhibits insulin signaling through the insulin receptor. the binding protein also binds insulin-like growth factors i and ii, proinsulin ... | 2000 | 10748036 |
| phosphorylation of the insect immunophilin fkbp46 by the spodoptera frugiperda homolog of casein kinase ii. | immunophilins are a family of conserved proteins found in both prokaryotes and eukaryotes, that exhibit peptidylprolyl isomerase (ppiase) activity. members of this family bind to immunosuppressive drugs and on this basis are divided into two classes: fkbps bind to fk506 and rapamycin, while cyclophilins bind to cyclosporin a. in this paper, we report on insect immunophilin fkbp46 and its associated kinase. the insect fkbp46 belongs to the high-molecular-weight immunophilins and shares many chara ... | 2000 | 10767538 |
| expression of the green fluorescent protein carried by autographa californica multiple nucleopolyhedrovirus in insect cell lines. | a recombinant acmnpv containing the green fluorescent protein (gfp) gene under the polyhedrin promoter (polh) was used to investigate the expression of the gfp gene as well as the production of recombinant extracellular virus in 14 continuous insect cell lines, including heliothis virescens (bcirl-hv-am1), helicoverpa zea (bcirl-hz-am1), anticarsia gemmatalis (bcirl-ag-am1), trichoplusia ni (tn-cl1), spodoptera frugiperda (iplb-sf21), spodoptera exigua (bcirl/amcy-se-e1 and bcirl/amcy-se-e5), bo ... | 2000 | 10777062 |
| human 5-hydroxytryptamine(5a) receptors activate coexpressed g(i) and g(o) proteins in spodoptera frugiperda 9 cells. | the ability of the human 5-hydroxytryptamine serotonin type 5a (h5-ht(5a)) receptor to couple to g proteins from distinct families was investigated through the simultaneous infection of spodoptera frugiperda 9 insect cells with recombinant baculoviruses encoding the various proteins. expression of g proteins was demonstrated in immunoblots. receptor-g protein coupling was monitored by high-affinity agonist binding and agonist-induced stimulation of [(35)s]guanosine-5'-o-(3-thio) triphosphate bin ... | 2000 | 10779389 |
| regulation of vitellogenesis in the fall armyworm, spodoptera frugiperda (lepidoptera: noctuidae). | studies were undertaken to investigate vitellogenesis and its regulation in female adults of the fall armyworm, spodoptera frugiperda. a single female-specific protein, likely to be the s. frugiperda vitellogenin (vg), appeared approximately 5 h after adult eclosion in the hemolymph of virgin females. the concentration of the protein increased with age as sodium dodecyl sulfate-polyacrylamide gel electrophoresis (sds-page) revealed. a protein with the same relative molecular mass was also presen ... | 2000 | 10802110 |
| high level expression of "male specific" pheromone binding proteins (pbps) in the antennae of female noctuiid moths. | pheromone binding proteins (pbps) are one branch of a multigene family of lepidopteran odorant binding proteins (obps) that are known for their relatively high levels of expression in male antennae. however, pbp expression has been observed at low levels in female antennae of the saturniidae, bombycidae and lymantriidae, and at relatively high levels in members of the noctuiidae. the function of female pbp expression is unclear, as female lepidoptera are consistently noted for their failure to r ... | 2000 | 10802242 |
| protein requirements for assembly of virus-like particles of junonia coenia densovirus in insect cells. | the coding sequences of four overlapping polypeptides starting at four different in-frame aug codons and co-terminating at the stop codon of the cap gene of junonia coenia densovirus (jcdnv) were inserted under the control of the p10 promoter of autographa californica nucleopolyhedrovirus (acmnpv) to generate acmnpv-vp1 (four polypeptides), acmnpv-vp2 (three polypeptides), acmnpv-vp3 (two polypeptides), and acmnpv-vp4 (one polypeptide) recombinant viruses. in all cases, infection of spodoptera f ... | 2000 | 10811945 |
| kinetic properties of the channels formed by the bacillus thuringiensis insecticidal crystal protein cry1c in the plasma membrane of sf9 cells. | spectrofluorimetric measurements were conducted to quantify, in real-time, membrane permeability changes resulting from the treatment of sf9 insect cells (spodoptera frugiperda, lepidoptera) with different bacillus thuringiensis cry insecticidal proteins. coumarin-derived cd222 and merocyanin-540 probes were respectively used to monitor extracellular k(+) and membrane potential variations upon sf9 cells incubation with cry toxins. our results establish that cry1c induces, after a delay, the depo ... | 2000 | 10811973 |
| determination of nadh-dependent glutamate synthase (gogat) in spodoptera frugiperda (sf9) insect cells by a selective 1h/15n nmr in vitro assay. | this is the second of two papers [drews, m., doverskog, m., ohman, l., chapman, b.e., jacobsson, u., kuchel, p.w., häggström, l., 2000. pathways of glutamine metabolism in spodoptera frugiperda (sf9) insect cells: evidence for the presence of the nitrogen assimilation system, and a metabolic switch by 1h/15n nmr. j. biotechnol. 78, 23-37]. where the general goal has been to determine and characterise the glutamine metabolism in sf9 cells. the presence of glutamate synthase (gogat) activity was i ... | 2000 | 10817344 |
| growth inhibitory effects on fall armyworm spodoptera frugiperda of some limonoids isolated from cedrela spp. (meliaceae). | dichloromethane extracts of cedrela salvadorensis and cedrela dugessi afforded a photogedunin epimeric mixture, gedunin and cedrelanolide. these compounds and the photogedunin epimeric acetates 3 and 4 at the 23-oh position were evaluated against spodoptera frugiperda. toosendanin, isolated from melia azedarach, was used as a positive control. when tested for activity on neonate larvae into the no-choice bioassays, gedunin, photogedunin epimeric mixture, and photogedunin acetates mixture caused ... | 2000 | 10820113 |
| influence of whorl region from resistant and susceptible corn genotypes on fall armyworm (lepidoptera: noctuidae) growth and development. | the effect of diets prepared from whorl tissue of resistant and susceptible corn genotypes, zea mays l., on the larval growth, development, and physiology of fall armyworm, spodoptera frugiperda (j. e. smith), was analyzed. larvae reared on an optimized artificial diet had a higher growth rate and developed faster than those reared on lyophilized whorl tissue from resistant and susceptible genotypes. larvae reared on the resistant material were smaller and had a longer developmental period. larv ... | 2000 | 10826203 |
| effects of mdr1 and mdr3 p-glycoproteins, mrp1, and bcrp/mxr/abcp on the transport of (99m)tc-tetrofosmin. | multidrug resistance (mdr1) p-glycoprotein (pgp), multidrug resistance-associated protein (mrp1), and breast cancer resistance protein (bcrp/mxr/abcp) are members of the atp-binding-cassette (abc) superfamily of membrane transporters and are thought to function as energy-dependent efflux pumps of a variety of structurally diverse chemotherapeutic agents. we herein report the characterization of (99m)tc-tetrofosmin, a candidate radiopharmaceutical substrate of abc transporters. (99m)tc-tetrofosmi ... | 2000 | 10856437 |
| filamentous actin is required for lepidopteran nucleopolyhedrovirus progeny production. | autographa californica m nucleopolyhedrovirus (acmnpv) is the prototypical member of the nucleopolyhedrosis: genus of the baculoviridae:, a family of large, double-stranded dna viruses that are highly diverse. nucleocapsid morphogenesis of acmnpv and others in the nucleopolyhedrovirus: genus takes place within the nuclei of infected host cells. previously, we showed that filamentous actin (f-actin) is essential for this process to occur in acmnpv-infected cells, an unprecedented finding for a dn ... | 2000 | 10859396 |
| interaction between manduca sexta allatotropin and manduca sexta allatostatin in the fall armyworm spodoptera frugiperda. | a peptide that strongly stimulates juvenile hormone (jh) biosynthesis in vitro by the corpora allata (ca) was purified from methanolic brain extracts of adult spodoptera frugiperda. using hplc separation followed by edman degradation and mass spectrometry, the peptide was identified as manduca sexta allatotropin (mas-at). treating the ca from adult s. frugiperda with synthetic mas-at (at 10(-6) m) caused an up to sevenfold increase in jh biosynthesis. the stimulation of jh synthesis was dose-dep ... | 2000 | 10876115 |
| stable gaba(a) receptor intermediates in sf-9 cells expressing alpha1, beta2 and gamma2 subunits. | spodoptera frugiperda insect cells (sf-9 cells) were used to study gaba(a) receptor assembly. time courses of the expression level of alpha1beta2 and alpha1beta2gamma2 receptor protein showed [(3)h]muscimol binding to appear 2 hr before [(3)h]flunitrazepam and [(35)s]tbps binding. this indicates that muscimol may bind to pentamers with an immature conformation or to molecules smaller than the pentamer. binding studies performed on fractions from sucrose gradients loaded with solubilized alpha1be ... | 2000 | 10878592 |
| mobility assays confirm the broad host-range activity of the minos transposable element and validate new transformation tools. | fast and reliable methods for assessing the mobility of the transposable element minos have been developed. these methods are based on the detection of excision and insertion of minos transposons from and into plasmids which are co-introduced into cells. excision is detected by polymerase chain reaction (pcr) with appropriate primers. transposition is assayed by marker rescue in escherichia coli, using a transposon plasmid that carries a tetracycline resistance gene and a target plasmid carrying ... | 2000 | 10886410 |
| cloning and kinetics of expression of brucella abortus heat shock proteins by baculovirus recombinants. | in an effort to develop genetically engineered brucella abortus (ba) vaccines, the genes encoding heat shock proteins (hsps) groel, groes, and htra were cloned and expressed in the bac-to-bac baculovirus system, and the kinetics of protein expression were analyzed using various insect cell lines in suspension cultures, different cell densities in suspension cultures, multiplicities of infection and recombinant virus replication times. trichoplusia ni cells expressed only ba htra, but spodoptera ... | 2000 | 10889410 |
| a unique 33-kd cysteine proteinase accumulates in response to larval feeding in maize genotypes resistant to fall armyworm and other lepidoptera. | plants respond to insect feeding with a number of defense mechanisms. using maize genotypes derived from antiquan germ plasm that are resistant to lepidoptera, we have demonstrated that a unique 33-kd cysteine proteinase accumulates in the whorl in response to larval feeding. the abundance of the proteinase increased dramatically at the site of larval feeding after 1 hr of infestation and continued to accumulate for as long as 7 days. the 33-kd cysteine proteinase was most abundant in the yellow ... | 2000 | 10899972 |
| size limits of thiocarbamides accepted as substrates by human flavin-containing monooxygenase 1. | microsomes isolated from spodoptera frugiperda (sf)9 cells infected with human flavin-containing monooxygenase (fmo)1 recombinant baculovirus catalyzed the nadph- and o2-dependent oxidation of methimazole, thiourea, and phenylthiourea. however, there was no detectable activity with 1,3-diphenylthiourea or larger thiocarbamides. microsomes from control sf9 cells were devoid of methimazole or thiourea s-oxygenase activity. trimethylamine up to 1.0 mm had no detectable effect on the oxidation of 10 ... | 2000 | 10901713 |
| changes in midgut endopeptidase activity of spodoptera frugiperda (lepidoptera: noctuidae) are responsible for adaptation to soybean proteinase inhibitors. | the development of transgenic maize plants expressing soybean proteinase inhibitors could reduce the economic damage of one of the major maize pests in brazil, the fall armyworm, spodoptera frugiperda (j.e. smith, 1797). we examined the influence of soybean proteinase inhibitors on digestive enzyme properties and development of s. frugiperda larvae. the inhibition of trypsin and chymotrypsin activities in vitro by soybean proteinase inhibitors suggested that either kunitz (sbti) or bowman-birk ( ... | 2000 | 10902346 |
| conventional resistance of experimental maize lines to corn earworm (lepidoptera: noctuidae), fall armyworm (lepidoptera: noctuidae), southwestern corn borer (lepidoptera: crambidae), and sugarcane borer (lepidoptera: crambidae). | plant resistance is a useful component of integrated pest management for several insects that are economically damaging to maize, zea mays l. in this study, 15 experimental lines of maize derived from a backcross breeding program were evaluated for resistance to corn earworm, helicoverpa zea (boddie); fall armyworm, spodoptera frugiperda (j. e. smith); southwestern corn borer, diatraea grandiosella dyar; and sugarcane borer, diatraea saccharalis (f.). experimental line 100-r-3 was resistant in t ... | 2000 | 10902359 |
| differences in the post-translational modifications of human papillomavirus type 6b major capsid protein expressed from a baculovirus system compared with a vaccinia virus system. | virus-like particles (vlps) are being currently investigated in vaccines against viral infections in humans. there are different recombinant-protein-expression systems available for obtaining the necessary vlp preparation for vaccination. however, the differences in post-translational modifications of the recombinant proteins obtained and their differences in efficacy in eliciting an anti-viral response in vaccines are not well established. in this study we have compared the post-translational m ... | 2000 | 10918035 |
| infectivity, transmission and 16s rrna sequencing of a rickettsia, coxiella cheraxi sp. nov., from the freshwater crayfish cherax quadricarinatus. | a rickettsia-like organism isolated from infected, farm-reared cherax quadricarinatus was cultured in the yolk sac of developing chicken eggs, but could not be cultured in 3 continuous cell lines, bluegill fry (bf-2), fathead minnow (fhm), and spodoptera frugiperda (sf-9). the organism was confirmed by fulfilling koch's postulates as the aetiological agent of mortalities amongst c. quadricarinatus. when c. quadricarinatus was inoculated with the organism, mortality was 100% at 28 degrees c and 8 ... | 2000 | 10918979 |
| culture in the rotating-wall vessel affects recombinant protein production capability of two insect cell lines in different manners. | the production of recombinant secreted alkaline phosphatase protein in virally infected insect cells was studied in shaker flask and high aspect rotating-wall vessel (harv) culture. two commonly used cell lines, spodoptera frugiperda sf-9 (sf-9) and a nonaggregating isolate of the trichoplusia ni bti-tn-5b1-4 (tn-5b1-4) cell line, trichoplusia ni tn-5b1-4-na (tn-5b1-4-na), were used and monitored for 120-h postinfection. different responses to culture in the harv were seen in the two cell lines. ... | 2000 | 10949994 |
| abnormal formation of polyhedra resulting from a single mutation in the polyhedrin gene of autographa californica multicapsid nucleopolyhedrovirus. | a spontaneous mutant that produces a single abnormally large cubic polyhedron per infected cell was isolated from a polyhedra-positive recombinant autographa californica multicapsid nucleopolyhedrovirus (acmnpv). both wild-type and mutant virus produce two forms of virus particles, budded virions and occluded virions. however, occluded virions are not found within the polyhedra of cells infected with mutant virus, as with the wild-type virus. these large cubic polyhedra do not have the typical l ... | 2000 | 10963398 |
| inhibition of [(3)h]ponasterone a binding by ecdysone agonists in the intact sf-9 cell line. | ecdysone agonists, including dibenzoylhydrazines, significantly inhibited the binding of [(3)h]ponasterone a ([(3)h]poa) in intact sf-9 cells (spodoptera frugiperda). the amount of [(3)h]poa binding varied in a concentration-dependent manner. according to the ic(50), concentration at which there is 50% inhibition, the order of potency of typical ecdysone agonists is tebufenozide (rh-5992) > methoxyfenozide (rh-2485) > poa > 20-hydroxyecdysone > cyasterone > rh-5849, makisterone a > or = inokoste ... | 2000 | 10978733 |
| the surface of prostate carcinoma du145 cells mediates the inhibition of urokinase-type plasminogen activator by maspin. | maspin is a novel serine protease inhibitor (serpin) with tumor suppressive potential in breast and prostate cancer, acting at the level of tumor invasion and metastasis. it was subsequently demonstrated that maspin inhibits tumor invasion, at least in part, by inhibiting cell motility. interestingly, in cell-free solutions, maspin does not inhibit several serine proteases including tissue-type plasminogen activator and urokinase-type plasminogen activator (upa). despite the recent biochemical e ... | 2000 | 10987285 |
| effects of long- and short-term passage of insect cells in different culture media on baculovirus replication. | two insect cell lines that had been maintained in both serum-free (sfm) and serum-containing (scm) media for over 5 years were each tested for their ability to replicate baculovirus. the gypsy moth cell line, iplb-ldeita (ld), produced similar (not statistically different) amounts of gypsy moth nucleopolyhedrovirus (ldmnpv) occlusion bodies (obs) in the two media (serum-free ex-cell 400 and tc-100 with 9% (v/v) fetal bovine serum, scm(1)) but produced more of the autographa californica nucleopol ... | 2000 | 11023743 |
| osmolarity effects on observed insect cell size after baculovirus infection are avoided using growth medium for sample dilution. | rates of cell size increase are an important measure of success during the baculovirus infection process. batch and fed batch cultures sustain large fluctuations in osmolarity that can affect the measured cell volume if this parameter is not considered during the sizing protocol. where osmolarity differences between the sizing diluent and the culture broth exist, biased measurements of size are obtained as a result of the cell osmometer response. spodoptera frugiperda (sf9) cells are highly sens ... | 2000 | 11027170 |
| on-line monitoring of physiological parameters of insect cell cultures during the growth and infection process. | on-line monitoring of insect cell cultures used for the production of recombinant proteins with the baculovirus expression vector system (bevs) provides valuable tools for the optimization, operation, and control of the production process. the relative permittivity (epsilon') and co(2) evolution rates (cer) were measured on-line using the biomass monitor and the infrared co(2) analyzer, respectively. the growth and infection phases of two different cell lines, spodoptera frugiperda (sf-9) and tr ... | 2000 | 11027174 |
| cell cycle progression in serum-free cultures of sf9 insect cells: modulation by conditioned medium factors and implications for proliferation and productivity. | cell cycle progression was studied in serum-free batch cultures of spodoptera frugiperda (sf9) insect cells, and the implications for proliferation and productivity were investigated. cell cycle dynamics in kbm10 serum-free medium was characterized by an accumulation of 50-70% of the cells in the g(2)/m phase of the cell cycle during the first 24 h after inoculation. following the cell cycle arrest, the cell population was redistributed into g(1) and in particular into the s phase. maximum rate ... | 2000 | 11027179 |
| rgs7 is palmitoylated and exists as biochemically distinct forms. | regulator of g protein signaling (rgs) proteins are gtpase-activating proteins that modulate neurotransmitter and g protein signaling. rgs7 and its binding partners galpha and gbeta5 are enriched in brain, but biochemical mechanisms governing rgs7/galpha/gbeta5 interactions and membrane association are poorly defined. we report that rgs7 exists as one cytosolic and three biochemically distinct membrane-bound fractions (salt-extractable, detergent-extractable, and detergent-insensitive) in brain. ... | 2000 | 11032900 |
| heterologous expression of cyp2k1 and identification of the expressed protein (bv-cyp2k1) as lauric acid (omega-1)-hydroxylase and aflatoxin b1 exo-epoxidase. | lmc2 is the most abundant constitutively expressed hepatic cytochrome p450 found in sexually immature rainbow trout (onchorynchus mykiss) and is also the isozyme that activates the carcinogen aflatoxin b1 (afb1). this p450 has been cloned, sequenced, and designated as cyp2k1. the present report describes the heterologous expression of enzymatically active cyp2k1 (bv-cyp2k1) in baculovirus spodoptera frugiperda (sf9) insect cells and its catalytic and immunoreactivity characterization in comparis ... | 2000 | 11038153 |
| the inhibitors of apoptosis of epiphyas postvittana nucleopolyhedrovirus. | in this study, four inhibitor of apoptosis genes (iaps) in the genome of epiphyas postvittana nucleopolyhedrovirus (eppomnpv) that are homologous to iap-1, iap-2, iap-3 and iap-4 genes of other baculoviruses have been identified. all four iap genes were sequenced and the iap-1 and iap-2 genes were shown to be functional inhibitors of apoptosis. the iap-1, iap-2 and iap-3 genes contain two baculovirus apoptosis inhibitor repeat motifs and a c(3)hc(4) ring finger-like motif. the activity of the ia ... | 2000 | 11038395 |
| purification and properties of a beta-glycosidase purified from midgut cells of spodoptera frugiperda (lepidoptera) larvae. | two beta-glycosidases (bg) (mr 47,000 and mr 50,000) were purified from spodoptera frugiperda (lepidoptera: noctuidae) midguts. these two polypeptides associate or dissociate depending on the medium ionic strength. the mr 47,000 bg probably has two active sites. one of the putative active sites (cellobiase site) hydrolyses p-nitrophenyl beta-d-glucoside (npbetaglu) (79% of the total activity in saturated enzyme), cellobiose, amygdalin and probably also cellotriose, cellotetraose and cellopentaos ... | 2000 | 11044660 |
| phylogenetic position of the diadromus pulchellus ascovirus dna polymerase among viruses with large double-stranded dna genomes. | the ascoviridae: is a family of large double-stranded (ds) dna insect viruses that contains four species, the spodoptera frugiperda (sfav1), trichoplusia ni (tnav2), heliothis virescens (hvav3) and diadromus pulchellus (dpav4) ascoviruses. these are unique among insect viruses in that the primary means of transmission among their lepidopteran hosts is generally by being vectored mechanically by hymenopteran parasitoids. ascoviruses are similar in virion structure, but their relationships with th ... | 2000 | 11086137 |
| characterization of repetitive dna regions and methylated dna in ascovirus genomes. | the accompanying phylogenetic study of large double-stranded dna viruses based on their delta dna polymerase genes suggests that ascoviruses (family ascoviridae:) and iridoviruses (family iridoviridae:) are closely related and may share a common ancestor. this relationship was unexpected because of marked differences between these viruses. iridoviruses produce icosahedral virions and occur broadly among vertebrates and invertebrates, whereas ascoviruses typically produce reniform or bacilliform ... | 2000 | 11086138 |
| a new ascovirus from spodoptera exigua and its relatedness to the isolate from spodoptera frugiperda. | a new ascovirus was isolated from spodoptera exigua in indonesia and was tentatively assigned as a new species, spodoptera exigua ascovirus 5a (seav-5a) according to the present ictv ascovirus naming scheme based on dna restriction fragment length polymorphism (rflp), hybridization, formation of occlusion body, tissue tropism and host spectrum. seav-5a replicated primarily in the fat body of susceptible hosts. seav-5a could be transmitted to s. frugiperda, pseudoplusia includens and trichoplusia ... | 2000 | 11086139 |
| co-expression of the capsid proteins of cowpea mosaic virus in insect cells leads to the formation of virus-like particles. | the regions of rna-2 of cowpea mosaic virus (cpmv) that encode the large (l) and small (s) coat proteins were expressed either individually or together in spodoptera frugiperda (sf21) cells using baculovirus vectors. co-expression of the two coat proteins from separate promoters in the same construct resulted in the formation of virus-like particles whose morphology closely resembled that of native cpmv virions. no such particles were formed when the individual l and s proteins were expressed. s ... | 2000 | 11086140 |
| crystallization of retinol dehydratase from spodoptera frugiperda: improvement of crystal quality by modification by ethylmercurythiosalicylate. | retinol dehydratase is a sulfotransferase which is presumed to catalyze the dehydration of its substrate via a transient retinyl sulfate intermediate. crystals (space group p2(1), unit-cell parameters a = 82.05, b = 66.61, c = 84.90 a, beta = 111.29 degrees ) are significantly improved by covalent modification of the protein with ethylmercury. | 2000 | 11092933 |
| identification of a novel isoform of the cyclic-nucleotide phosphodiesterase pde3a expressed in vascular smooth-muscle myocytes. | we have identified a new cyclic-nucleotide phosphodiesterase isoform, pde3a, and cloned its cdna from cultured aortic myocytes. the nucleotide sequence of its coding region is similar to that of the previously cloned myocardial isoform except for the absence of the initial 300-400 nt that are present in the latter, as confirmed by reverse-transcriptase-mediated pcr, 5' rapid amplification of cdna ends and a ribonuclease protection assay. expression in spodoptera frugiperda (sf9) cells yields a p ... | 2001 | 11115397 |
| adenine excisional repair function of myh protein on the adenine:8-hydroxyguanine base pair in double-stranded dna. | adenine paired with 8-hydroxyguanine (oh(8)g), a major component of oxidative dna damage, is excised by myh base excision repair protein in human cells. since repair activity of myh protein on an a:g mismatch has also been reported, we compared the repair activity of his(6)-tagged myh proteins, expressed in spodoptera frugiperda sf21 cells, on a:oh(8)g and a:g mismatches by dna cleavage assay and gel mobility shift assay. we also compared the repair ability of type 1 mitochondrial protein with t ... | 2000 | 11121482 |
| the xenobiotic inhibitor profile of cytochrome p4502c8. | to investigate inhibition of recombinant cyp2c8 by: (i) prototypic cyp isoform selective inhibitors (ii) imidazole/triazole antifungal agents (known inhibitors of cyp), and (iii) certain cyp3a substrates (given the apparent overlapping substrate specificity of cyp2c8 and cyp3a). | 2000 | 11136296 |
| effect of two granulosis viruses on the activity of the gypsy moth (lepidoptera: lymantriidae) nuclear polyhedrosis virus. | two granulosis viruses (gv) were tested as enhancers for the gypsy moth nuclear polyhedrosis virus (ldmnpv). helicoverpa armigera (hübner) cv (hagv) had no detrimental effect upon larval growth and development, but in combination with ldmnpv it reduced both the lc50 and the lt50 for the npv. in addition, the combination also adversely affected the growth and development of gypsy moth larvae. the lc50 of ldmnpv was reduced by as much as 300-fold (hagv at 10(-2) dilution) and the lt50 was reduced ... | 2000 | 11142292 |
| production of biologically active recombinant bovine interferon-gamma by two different baculovirus gene expression systems using insect cells and silkworm larvae. | the full-length bovine interferon-gamma (bifn-gamma) cdna, including the secretion signal peptide coding region was recloned into baculovirus transfer vectors pacym1 and pbm050. these vectors were co-transfected with autographa californica nuclear polyhedrosis virus (acnpv) or bombyx mori nuclear polyhedrosis virus (bmnpv) dna into spodoptera frugiperda cells (sf21ae) and bombyx mori cells (bmn), respectively. the recombinant viruses, named acbifn-gamma and bmbifn-gamma, were then recovered. rec ... | 2001 | 11145838 |
| comparison of the biological activities of human immunodeficiency virus 1 p24 and gp41 expressed in spodoptera frugiperda cells by use of bac-to-bac system. | recombinant transposing plasmids pfh24 and pfh41 were constructed by cloning the human immunodeficiency virus 1 (hiv-1) p24 and gp41 genes, respectively, into the transposing vector pfastbachta. recombinant bacmids rbh24 and rbh41 were obtained by transposing ppolh/p24 and ppolh/gp41 expression cassettes from recombinant plasmids pfh24 and pfh41, respectively. recombinant viruses rach24 and rach41 were generated by transfection of the spodoptera frugiperda (sf9) cells with the dnas of plasmids r ... | 2000 | 11155353 |
| interaction of hydrophobic anions with the rat skeletal muscle chloride channel clc-1: effects on permeation and gating. | permeation of a range of hydrophobic anions through the rat skeletal muscle chloride channel, rclc-1, expressed in sf-9 (a spodoptera frugiperda insect cell line) cells has been studied using the whole-cell patch-clamp technique. bi-ionic reversal potentials measured with external application of foreign anions gave the following permeability sequence: cl- (1) > benzoate (0.15) > hexanoate (0.12) > butyrate (0.09) > propionate (0.047) approximately formate (0.046). anions with larger hydrophobic ... | 2001 | 11158270 |
| production of functional antibodies generated in a nonlytic insect cell expression system. | a monoclonal antibody directed against the type 2 adenovirus (ad2) penton base protein was cloned and expressed in spodoptera frugiperda (sf9) cells using a nonlytic vector system. the coding sequences for the immunoglobulin light and heavy chains were placed under the control of the orgyia pseudotsugata multicapsid nucleopolyhedrosis virus immediate-early 2 (opie2) promoter. transfected sf9 cells continuously secreted the antibody which retained the ability to recognize both native and recombin ... | 2001 | 11162396 |
| high-level expression of human acidic fibroblast growth factor and basic fibroblast growth factor in silkworm (bombyx mori l.) using recombinant baculovirus. | a hybrid of autographa californica nuclear polyhedrosis virus and bombyx mori nuclear polyhedrosis virus, which is infectious to both spodoptera frugiperda and bombyx mori, was prepared in our previous study. two recombinant hybrid baculoviruses, carrying cdnas of human acidic and basic fibroblast growth factors, respectively, were successfully constructed in this study, for the large-scale production of human afgf and bfgf using silkworm as host. these recombinant viruses were used to inoculate ... | 2001 | 11162406 |
| alpha-galactosidases from the larval midgut of tenebrio molitor (coleoptera) and spodoptera frugiperda (lepidoptera). | there are three midgut alpha-galactosidases (tg1, tg2, tg3) from tenebrio molitor larvae that are partially resolved by ion-exchange chromatography. the enzymes have approximately the same ph optimum (5.0), pl value (4.6) and mr value (46000-49000) as determined by gel filtration or native electrophoresis run in polyacrylamide gels with different concentrations. substrate specificities and functions were proposed for the major t. molitor midgut alpha-galactosidases (tg2 and tg3) based on chromat ... | 2001 | 11163310 |
| 5-hydroxytryptamine 4(a) receptor expressed in sf9 cells is palmitoylated in an agonist-dependent manner. | the mouse 5-hydroxytryptamine 4(a) receptor [5-ht(4(a))] was expressed with a baculovirus system in insect cells and analysed for acylation. [(3)h]palmitic acid was effectively incorporated into 5-ht(4(a)) and label was sensitive to the treatment with reducing agents indicating a thioester-type bond. analysis of protein-bound fatty acids revealed that 5-ht(4(a)) contains predominantly palmitic acid. treatment of infected sf9 (spodoptera frugiperda) cells with bimu8 [(endo-n-8-methyl-8-azabicyclo ... | 2001 | 11171060 |
| expression and characterization of recombinant human alpha-3/4-fucosyltransferase iii from spodoptera frugiperda (sf9) and trichoplusia ni (tn) cells using the baculovirus expression system. | the human alpha-3/4-fucosyltransferase iii (fuc-tiii) participates in the synthesis of lewis determinants. the enzyme from human sources is scarce and heterogeneous. in this paper we describe the expression of a secreted form of fuc-tiii (sft3) in two insect cell lines, spodoptera frugiperda (sf9) and trichoplusia ni (tn), using the baculovirus expression system. the sf9 cells secreted approx. 0.4 unit/l (1 mg/l) of the enzyme. the tn cells secreted approx. 3-fold this amount. a large proportion ... | 2001 | 11171070 |
| characterization and mapping of the 12 kda fk506-binding protein (fkbp12)-binding site on different isoforms of the ryanodine receptor and of the inositol 1,4,5-trisphosphate receptor. | we investigated the interaction of the 12 kda fk506-binding protein (fkbp12) with two ryanodine-receptor isoforms (ryr1 and ryr3) and with two myo-inositol 1,4,5-trisphosphate (ip3) receptor isoforms (ip3r1 and ip3r3). using glutathione s-transferase (gst)-fkbp12 affinity chromatography, we could efficiently extract ryr1 (42+/-7% of the solubilized ryr1) from terminal cisternae of skeletal muscle as well as ryr3 (32+/-4% of the solubilized ryr3) from ryr3-overexpressing hek-293 cells. these inte ... | 2001 | 11171121 |
| expression of scavenger receptor class b type 1 (sr-bi) promotes microvillar channel formation and selective cholesteryl ester transport in a heterologous reconstituted system. | in the "selective" cholesteryl ester (ce) uptake process, surface-associated lipoproteins [high density lipoprotein (hdl) and low density lipoprotein] are trapped in the space formed between closely apposed surface microvilli (microvillar channels) in hormone-stimulated steroidogenic cells. this is the same location where an hdl receptor (sr-bi) is found. in the current study, we sought to understand the relationship between sr-bi and selective ce uptake in a heterologous insect cell system. sf9 ... | 2001 | 11171999 |
| correlating differences in larval survival and development of bollworm (lepidoptera: noctuidae) and fall armyworm (lepidoptera: noctuidae) to differential expression of cry1a(c) delta-endotoxin in various plant parts among commercial cultivars of transgenic bacillus thuringiensis cotton. | differences in larval survival and development of bollworm, helicoverpa zea (boddie), and fall armyworm, spodoptera frugiperda (j. e. smith), respectively, were found to exist among commercially available cry1a(c) transgenic bacillus thuringiensis berliner (bt) varieties. using a quantification assay (elisa) to measure the levels of delta-endotoxin in two of these varieties ('dp 451b/rr' and 'nucotn 33b'), differences in the amount of delta-endotoxin present in various plant parts was correlated ... | 2001 | 11233127 |
| tyr199 in transmembrane domain 5 of the beta2-adrenergic receptor interacts directly with the pharmacophore of a unique fluorenone-based antagonist. | mutagenesis of the beta2-adrenergic receptor (beta2ar) has suggested that amino acids in transmembrane domain 5 (tmd 5) play an important role in the interaction of the receptor with the catechol end of adrenergic agonists. however, little direct biochemical evidence for the interaction of any beta2ar agonist or antagonist with tmd 5 has been reported. to identify receptor amino acids that contribute to the beta2ar antagonist binding site, we identified the precise amino acid photoinsertion site ... | 2001 | 11237852 |
| comparison of escherichia coli, saccharomyces cerevisiae, pichia pastoris, spodoptera frugiperda, and cos7 cells for recombinant gene expression. application to a rabbit liver carboxylesterase. | expression of a rabbit liver carboxylesterase has been achieved in several different model systems including escherichia coli, pichia pastoris, saccharomyces cerevisiae, spodoptera frugiperda, and cos7 cells. although, recombinant protein was observed in e. coli sonicates, little or no enzymatic activity was detected. similarly, no activity was observed following expression in s. cerevisiae. in contrast, active protein was produced in p. pastoris, from s. frugiperda following baculoviral infecti ... | 2000 | 11252804 |
| selective recognition of inositol phosphates by subtypes of the inositol trisphosphate receptor. | synthetic analogues of inositol trisphosphate (ip(3)), all of which included structures equivalent to the 4,5-bisphosphate of (1,4,5)ip(3), were used to probe the recognition properties of rat full-length type 1, 2 and 3 ip(3) receptors expressed in insect spodoptera frugiperda 9 cells. using equilibrium competition binding with [(3)h](1,4,5)ip(3) in ca(2+)-free cytosol-like medium, the relative affinities of the receptor subtypes for (1,4,5)ip(3) were type 3 (k(d)=11+/-2 nm)>type 2 (k(d)=17+/-2 ... | 2001 | 11256949 |
| modulation of inositol 1,4,5-trisphosphate binding to the various inositol 1,4,5-trisphosphate receptor isoforms by thimerosal and cyclic adp-ribose. | three different genes encode the inositol 1,4,5-trisphosphate (ip3) receptor (ip3r), an intracellular ca2+ channel involved in cellular ca2+ signaling. the ip3-binding characteristics of the various ip3r isoforms differ, but until now no specific activators or inhibitors of ip3 binding have been described. we compared the effects of oxidizing reagents, in particular thimerosal, and of cyclic adp-ribose (cadpr) on ip3 binding to the various ip3r isoforms. we therefore expressed the n-terminal 581 ... | 2001 | 11274965 |
| potent antiviral potamogetonyde and potamogetonol, new furanoid labdane diterpenes from potamogeton malaianus. | new furanoid labdane diterpenes, potamogetonyde (3) and potamogetonol (4), together with two known compounds, potamogetonin (1) and 15,16-epoxy-12-oxo-8(17),13(16),14-labdatrien-20,19-olide (2), were isolated from the ch(2)cl(2) extract of potamogeton malaianus. the chemical structures of 1-4 were elucidated by the analyses of their spectral data, mainly by 1d and 2d nmr techniques. potamogetonyde (3) and potamogetonol (4) exhibited potent antiviral (hsv-1) activity with respective ic(50) values ... | 2001 | 11277765 |
| apoptosis in motion. an apical, p35-insensitive caspase mediates programmed cell death in insect cells. | activation of caspases by proteolytic processing is a critical step during apoptosis in metazoans. here we use high resolution time lapse microscopy to show a tight link between caspase activation and the morphological events delineating apoptosis in cultured sf21 cells from the moth spodoptera frugiperda, a model insect system. the principal effector caspase, sf-caspase-1, is proteolytically activated during sf21 apoptosis. to define the potential role of initiator caspases in vivo, we tested t ... | 2001 | 11278634 |
| molecular determinants of desensitization and assembly of the chimeric gaba(a) receptor subunits (alpha1/gamma2) and (gamma2/alpha1) in combinations with beta2 and gamma2. | two gamma-aminobutyric acid(a) (gaba(a)) receptor chimeras were designed in order to elucidate the structural requirements for gaba(a) receptor desensitization and assembly. the (alpha1/gamma2) and (gamma2/alpha1) chimeric subunits representing the extracellular n-terminal domain of alpha1 or gamma2 and the remainder of the gamma2 or alpha1 subunits, respectively, were expressed with beta2 and beta2gamma2 in spodoptera frugiperda (sf-9) cells using the baculovirus expression system. the (alpha1/ ... | 2001 | 11290383 |
| novel sp family-like transcription factors are present in adult insect cells and are involved in transcription from the polyhedrin gene initiator promoter. | we earlier documented the involvement of a cellular factor, polyhedrin (polh) promoter-binding protein, in transcription from the autographa californica nuclear polyhedrosis virus polh gene promoter. sequences upstream of the polh promoter were found to influence polh promoter-driven transcription. analysis of one such region, which could partially compensate for the mutated polh promoter and also activate transcription from the wild-type promoter, revealed a sequence (acsp) containing a caccc m ... | 2001 | 11294840 |
| affinity purification of secreted alkaline phosphatase produced by baculovirus expression vector system. | human secreted alkaline phosphatase (seap) was produced in a stably-transformed spodoptera frugiperda sf-9 insect cell line (sfb4galt) following infection with a recombinant autographa californica multiple nuclear polyhedrovirus containing the seap gene under control of the polyhedrin promoter. an affinity chromatographic column prepared by linking 4-amino-benzylphosphonic acid to histidyl-expoxy-sepharose was used to isolate seap from the cell supernatant following removal of cells and virus an ... | 2001 | 11297388 |
| insect growth regulatory activity of some extracts and compounds from parthenium argentatum on fall armyworm spodoptera frugiperda. | the methanolic extract from aerial parts of parthenium argentatum, afforded argentatin a and b. these compounds were evaluated for their effect on the fall armyworm (spodoptera frugiperda). toosendanin, a commercial insecticide derived from melia azedarach was used as positive control. when tested for activity, using neonate larvae into the no-choice artificial diet bioassays, argentatin a, argentatin b and methanol extract caused significant growth inhibitory activity with gc50 of 17.8, 36.1 an ... | 2001 | 11302221 |
| defining mechanisms of toxicity for linoleic acid monoepoxides and diols in sf-21 cells. | linoleic acid monoepoxides have been correlated with many pathological conditions. studies using insect cells derived from spodoptera frugiperda (sf-21 cells) have suggested that conversion of the epoxides to the diols is required for toxicity. however, more recent studies using rabbit renal proximal tubules have suggested that linoleic acid monoepoxides are direct mitochondrial toxins. to better understand these discrepancies, we compared the toxicity of these linoleic acid metabolites in sf-21 ... | 2001 | 11304132 |
| association of pcr and feeding bioassays as a large-scale method to screen tropical bacillus thuringiensis isolates for a cry constitution with higher insecticidal effect against spodoptera frugiperda (lepidoptera: noctuidae) larvae. | to verify whether the presence of any of the cry1c, 1d, 1e and 1f genes could be associated with high toxicity against fall armyworm. | 2001 | 11328507 |
| role of glycine-534 and glycine-1179 of human multidrug resistance protein (mdr1) in drug-mediated control of atp hydrolysis. | the human multidrug resistance protein (mdr1) (p-glycoprotein), a member of the atp-binding cassette (abc) family, causes multidrug resistance by an active transport mechanism, which keeps the intracellular level of hydrophobic compounds below a cell-killing threshold. human mdr1 variants with mutations affecting a conserved glycine residue within the abc signature of either or both abc units (g534d, g534v, g1179d and g534d/g1179d) were expressed and characterized in spodoptera frugiperda (sf9) ... | 2001 | 11336637 |
| cloning and characterization of an inhibitor of apoptosis protein (iap) from bombyx mori. | we cloned a novel inhibitor of apoptosis protein (iap) family member, bmiap, from bombyx mori bmn cells. bmiap contains two baculoviral iap repeat (bir) domains followed by a ring domain. bmiap shares striking amino acid sequence similarity with lepidopteran iaps, sfiap and tniap, and with two baculoviral iaps, cpiap and opiap, suggesting evolutionary conservation. bmiap blocks programmed cell death (apoptosis) in spodoptera frugiperda sf-21 cells induced by p35 deficient autographa californica ... | 2001 | 11341966 |
| amino acid residues involved in substrate binding and catalysis in an insect digestive beta-glycosidase. | a beta-glycosidase (m(r) 50000) from spodoptera frugiperda larval midgut was purified, cloned and sequenced. it is active on aryl and alkyl beta-glucosides and cellodextrins that are all hydrolyzed at the same active site, as inferred from experiments of competition between substrates. enzyme activity is dependent on two ionizable groups (pk(a1)=4.9 and pk(a2)=7.5). effect of ph on carbodiimide inactivation indicates that the pk(a) 7.5 group is a carboxyl. k(cat) and k(m) values were obtained fo ... | 2001 | 11342030 |
| characterization of cytochrome p450 2a4 and 2a5-catalyzed 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (nnk) metabolism. | the tobacco-specific nitrosamine, 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (nnk), is a potent lung carcinogen in the a/j mouse, and is believed to be a causative agent for human lung cancer. nnk requires metabolic activation by alpha-hydroxylation to exert its carcinogenic potential. the human p450, 2a6 is a catalyst of this reaction. there are two closely related enzymes in the mouse, p450 2a4 and 2a5, which differ from each other by only 11 amino acids. in the present study these two mou ... | 2000 | 11368333 |
| antioxidant defense systems of two lipidopteran insect cell lines. | spodoptera frugiperda sf-9 (sf-9) and trichoplusia ni bti-tn-5b1-4 (tn-5b1-4) insect cell lines were found to contain unique assemblages of antioxidant enzymes. specifically, the sf-9 insect cell line contained manganese and copper-zinc superoxide dismutase (mnsod and cuznsod) for reducing the superoxide radical (o(2)(*-)) to hydrogen peroxide (h(2)o(2)) and ascorbate peroxidase (apox) for reducing the resulting h(2)o(2) to h(2)o. approximately one third of the total sod activity was found to be ... | 2001 | 11368923 |
| determination of nucleotides and sugar nucleotides involved in protein glycosylation by high-performance anion-exchange chromatography: sugar nucleotide contents in cultured insect cells and mammalian cells. | we have developed a simple and highly sensitive hplc method for determination of cellular levels of sugar nucleotides and related nucleotides in cultured cells. separation of 9 sugar nucleotides (cmp-neu5ac, cmp-neu5gc, cmp-kdn, udp-gal, udp-glc, udp-galnac, udp-glcnac, gdp-fuc, gdp-man) and 12 nucleotides (amp, adp, atp, cmp, cdp, ctp, gmp, gdp, gtp, ump, udp, and utp) was examined by reversed-phase hplc and high-performance anion-exchange chromatography (hpaec). although the reversed-phase hpl ... | 2001 | 11373089 |
| the peritrophic membrane of spodoptera frugiperda: secretion of peritrophins and role in immobilization and recycling digestive enzymes. | a peritrophin from the spodoptera frugiperda peritrophic membrane (pm) and microvillar proteins from s. frugiperda anterior midgut cells were isolated and used to raise antibodies in a rabbit. these antibodies, as well as a tenebrio molitor amylase antibody that cross-reacts with s. frugiperda amylases, and wheat-germ aglutinin were used in immunolocalization experiments performed with the aid of confocal fluorescence and immunogold techniques. the results showed that the peritrophin was secrete ... | 2001 | 11376453 |
| expression in spodoptera frugiperda (sf21) insect cells of bt-r(1), a cadherin-related receptor from manduca sexta for bacillus thuringiensis cry1ab toxin. | the cadherin-related receptor of manduca sexta, bt-r(1), for the cry1a family of bacillus thuringiensis insecticidal toxins, was expressed in cultured spodoptera frugiperda (sf21) insect cells utilizing the expression vector deltaop-gp64. recombinant bt-r(1) was released by the sf21 cells in soluble form into the culture medium and represents approximately 58% of total bt-r(1) produced by the cells. the soluble protein was purified by affinity chromatography using cry1ab toxin coupled to sepharo ... | 2001 | 11388812 |
| recombinant expression and purification of human androgen receptor in a baculovirus system. | a full-length human androgen receptor (har) cdna was used to produce recombinant baculovirus. spodoptera frugiperda (sf9) cells infected with this virus expressed protein with an n-terminal hexahistidine tag (his(6)-har) in soluble and insoluble forms. the soluble cytosolic his(6)-har demonstrated similar association and dissociation half-times for mibolerone, similar binding affinity for mibolerone, and similar steroid specificity as bona fide ar. under native conditions, the soluble cytosolic ... | 2001 | 11396977 |
| establishment and characterization of cell-free translation/glycosylation in insect cell (spodoptera frugiperda 21) extract prepared with high pressure treatment. | a coupled cell-free translation/glycosylation system, prepared from spodoptera frugiperda insect cells, was established and optimized for protein production and glycosylation efficiency. both translation and glycosylation were stimulated by addition of mg2+, k+, atp, gtp, creatine kinase and creatine phosphate, suggesting that glycoprotein productivity is largely determined by translation efficiency. however, high concentrations of creatine phosphate significantly inhibited translation. spermidi ... | 2001 | 11398925 |
| typical antipsychotics exhibit inverse agonist activity at rat dopamine d1-like receptors expressed in sf9 cells. | the baculovirus system has been used to express the rat dopamine d1 receptors in spodoptera frugiperda (sf9) cells. a panel of typical antipsychotics including, alpha-flupenthixol, fluphenazine and thioridizine were found to inhibit dopamine-dependent stimulation of adenylyl cyclase. however, these compounds were also found to inhibit adenylyl cyclase activity in the absence of agonist in sf9 cells expressing dopamine d1-like receptors. therefore, these nonselective dopamine receptor compounds d ... | 2001 | 11408027 |
| nature of the oligomers formed by muscarinic m2 acetylcholine receptors in sf9 cells. | wild-type, flag-tagged, and c-myc-tagged muscarinic m2 receptors extracted in digitonin-cholate from singly and co-infected sf9 (spodoptera frugiperda) cells were indistinguishable in their binding of [3h]quinuclidinylbenzilate, either before or after purification. the flag epitope was found to coimmunoprecipitate with the c-myc epitope when co-infected cells were solubilised in digitonin-cholate, n-dodecyl-beta-d-maltoside or lubrol-px. the degree of coprecipitation in digitonin-cholate was una ... | 2001 | 11408044 |
| characterization of the cdna encoding the 90 kda heat-shock protein in the lepidoptera bombyx mori and spodoptera frugiperda. | this report presents the first hsp90 complete cdna sequences from two lepidoptera. the bombyx mori full sequence was reconstituted from 15 partial cdna clones belonging to expressed sequence tag libraries obtained from different tissues or cultured cells, thus showing the ubiquitous expression of the hsp90 gene. the spodoptera frugiperda cdna was isolated as a full-length clone from a cdna library established from the sf9 cell line. both cdnas are highly homologous and display the classical amin ... | 2001 | 11418243 |
| acetylcholinesterase and insect growth inhibitory activities of gutierrezia microcephala on fall armyworm spodoptera frugiperda j.e. smith. | from the aerial parts of gutierrezia microcephala (asteraceae), four oxyflavones were isolated, namely 5,7,2'-trihydroxy-3,6,8,4',5'-pentamethoxyflavone (1); 5,7,4'-trihydroxy-3,6,8-trimethoxyflavone (2); 5,7,2',4'-tetrahydroxy-3,6,8,5'-tetramethoxyflavone (3); 5,2'-dihydroxy-3,6,7,8,4',5'-hexamethoxyflavone (4), and an ent-clerodane, bacchabolivic acid (5). compounds 1-5, the synthetic methyl ester (6), n-hexane and meoh extracts were evaluated against the fall armyworm (spodoptera frugiperda). ... | 2001 | 11421454 |