| comparison of field-collected ascovirus isolates by dna hybridization, host range, and histopathology. | six field-collected ascovirus isolates obtained from five noctuid species in the continental united states were compared with respect to the general relatedness of their dna, host range, and histopathology. two isolates were from spodoptera frugiperda, and the other four were from autographa precationis, heliothis virescens, helicoverpa zea, and trichoplusia ni. dna-dna hybridization studies showed that the six isolates belonged to three distinct viral species, with the isolates from s. frugiper ... | 1998 | 9709014 |
| high-level expression of the human cb2 cannabinoid receptor using a baculovirus system. | a human cb2 recombinant baculovirus (acnpv-hcb2) was generated by site-specific transposition and employed to express the human cb2 cannabinoid receptor. northern analysis of total rna from spodoptera frugiperda (sf9) insect cells infected with acnpv-hcb2 revealed novel expression of a unique 2.3 kb transcript when probed with hcb2 cdna. this transcript corresponded to the size expected for hcb2 generated from the recombinant virus construct. western immunoblot analysis of whole cell homogenates ... | 1998 | 9714308 |
| a novel role for calmodulin: ca2+-independent inhibition of type-1 inositol trisphosphate receptors. | calmodulin inhibits both inositol 1,4,5-trisphosphate (ip3) binding to, and ip3-evoked ca2+ release by, cerebellar ip3 receptors [patel, morris, adkins, o'beirne and taylor (1997) proc. natl. acad. sci. u. s.a. 94, 11627-11632]. in the present study, full-length rat type-1 and -3 ip3 receptors were expressed at high levels in insect spodoptera frugiperda 9 cells and the effects of calmodulin were examined. in the absence of ca2+, calmodulin caused a concentration-dependent and reversible inhibit ... | 1998 | 9716504 |
| expression and characterization of canine cytochrome p450 2d15. | cyp2d15 is the canine ortholog of human cyp2d6, the human cyp2d isoform involved in the metabolism of drugs such as antiarhythmics, adrenoceptor antagonists, and tricyclic antidepressants. similar to human, canine cyp2d15 is expressed in the liver, with detectable levels in several other tissues. three different cyp2d15 cdna clones were obtained by rt-pcr from dog liver rna. two clones corresponded to variant full-length cyp2d15 cdnas (termed cyp2d15 wt2 and cyp2d15 v1); the third was identified ... | 1998 | 9721180 |
| tyrosine structural changes detected during the photoactivation of rhodopsin. | we present the first fourier transform infrared (ftir) analysis of an isotope-labeled eukaryotic membrane protein. a combination of isotope labeling and ftir difference spectroscopy was used to investigate the possible involvement of tyrosines in the photoactivation of rhodopsin (rho). rho --> mii difference spectra were obtained at 10 degrees c for unlabeled recombinant rho and isotope-labeled l-[ring-2h4]tyr-rho expressed in spodoptera frugiperda cells grown on a stringent culture medium conta ... | 1998 | 9726981 |
| receptor cell habituation in the a1 auditory receptor of four noctuoid moths. | moths of both sexes of empyreuma affinis (=pugione) and syntomeida epilais (arctiidae, ctenuchinae), maenas jussiae (arctiidae, arctiinae) and spodoptera frugiperda (noctuidae, amphipyrinae) were studied. spike activity in the a1 cell was recorded using a stainless-steel hook electrode from the tympanic nerve in the mesothorax. acoustic stimuli consisting of 25 and 100 ms pulses at the best frequency for the species and at intensities that evoke a1 cell saturation response were used at repetitio ... | 1998 | 9739070 |
| baculovirus-mediated expression of recombinant rat phosphatidylcholine transfer protein. | phosphatidylcholine transfer protein catalyzes intermembrane transfer of phosphatidylcholines exclusive of all other phospholipid classes. although postulated to participate in phosphatidylcholine biosynthesis and biliary trafficking in liver, the molecular basis underlying the substrate specificity of phosphatidylcholine transfer protein remains to be elucidated. having demonstrated the inability of escherichia coli to express recombinant phosphatidylcholine transfer protein, we infected spodop ... | 1998 | 9741699 |
| functional properties of a naturally occurring isoform of soluble guanylyl cyclase. | soluble guanylyl cyclase (sgc), the target enzyme of the signalling molecule no, contains one prosthetic haem group and consists of an alpha and a beta subunit. so far, only the alpha1beta1 heterodimer has been shown to exist in different cells and tissues, and most biochemical studies of sgc have been performed with the alpha1 beta1 heterodimer. here we demonstrate for the first time the natural occurrence of the alpha2 subunit on the protein level. the alpha2 subunit co-precipitated with the b ... | 1998 | 9742221 |
| apoptosis resulting from superinfection of heliothis zea virus 1 is inhibited by p35 and is not required for virus interference. | superinfection of spodoptera frugiperda insect cells that are persistently infected with heliothis zea 1 (hz-1) virus induces general cellular apoptosis and subsequently results in homologous virus interference. since apoptosis correlates closely with both a significant decrease in yield of virus progeny and expansion of virus infection among cells, further experiments were designed to verify the direct association of apoptosis with homologous interference. it was found that superinfection-induc ... | 1998 | 9747740 |
| site-specific, enzymatic biotinylation of recombinant proteins in spodoptera frugiperda cells using biotin acceptor peptides. | site-specific, enzymatic biotinylation of recombinant proteins can be exploited to circumvent many problems associated with the use of biotinylating reagents in vitro and to overcome some of their inherent limitations. additionally, biotinyl proteins can be purified to near-homogeneity in a single step under native conditions. here we report that a biotin acceptor peptide (bap) substrate for escherichia coli biotin holoenzyme synthetase (bira) can be used to label recombinant proteins with bioti ... | 1998 | 9750126 |
| expression of recombinant human acid sphingomyelinase in insect sf21 cells: purification, processing and enzymatic characterization. | biochemical and structural studies on human acid sphingomyelinase (hasmase) depend on the access to homogeneous biologically active enzyme. due to the low abundance of native hasmase (n-hasmase) in human tissue, conventional purification strategies are not suitable for the isolation of preparative amounts of the enzyme. we describe a novel approach to the functional expression and purification of hasmase employing the baculovirus expression vector system. infection of spodoptera frugiperda 21 ce ... | 1998 | 9764481 |
| persistent baculovirus infection results from deletion of the apoptotic suppressor gene p35. | infection with the wild-type baculovirus autographa californica multiple nuclear polyhedrosis virus (acmnpv) results in complete death of spodoptera frugiperda (sf) cells. however, infection of sf cells with acmnpv carrying a mutation or deletion of the apoptotic suppressor gene p35 allowed the cloning of surviving sf cells that harbored persistent viral genomes. persistent infection established with the virus with p35 mutated or deleted was blocked by stable transfection of p35 in the host geno ... | 1998 | 9765462 |
| characterization of the human gonadotropin-releasing hormone receptor heterologously produced using the baculovirus/insect cell and the semliki forest virus systems. | 1. two eukaryotic viral systems, the baculovirus/insect cell and the semliki forest virus systems, were tested for heterologous expression of human gonadotropin-releasing hormone receptor (gnrhr) cdna. 2. an unmodified as well as a c-myc epitope-tagged human gnrh receptor was produced in two insect cell lines (spodoptera frugiperda, trichoplusia ni) after infection with the respective recombinant baculoviruses. in both insect cell lines, the receptor was identified by immunoblot analysis as a tr ... | 1998 | 9777251 |
| polyhedrin sequence determines the tetrahedral shape of occlusion bodies in thysanoplusia orichalcea single-nucleocapsid nucleopolyhedrovirus. | a nucleopolyhedrovirus (npv) isolated from the looper thysanoplusia orichalcea l. (lepidoptera: noctuidae) (thornpv) is occluded in a tetrahedral protein matrix. the orf of the thornpv polyhedrin gene contains 738 nt which code for 246 amino acids of the putative polyhedrin protein with an estimated molecular mass of 28,778 da. the promoter of this gene is similar in length to the promoter of spodoptera frugiperda npv (sfmnpv), with a 5 nt deletion before the start codon compared to those of oth ... | 1998 | 9780063 |
| expression of bovine activin-a and inhibin-a in recombinant baculovirus-infected spodoptera frugiperda sf 21 insect cells. | currently, bioactive activin and inhibin for investigative purposes are obtained either by purification from bovine or porcine follicular fluid or have been kindly supplied in limited amounts by genentech. the latter are recombinant formulations produced in cultured monkey kidney cv-1 cells. the aims of this study were to assess the potential of the baculovirus expression system as an alternative means to produce recombinant activin and inhibin. towards these goals, two recombinant baculoviruses ... | 1998 | 9785478 |
| nonenzymatic palmitoylation at cys 3 causes extra-activation of the alpha-subunit of the stimulatory gtp-binding protein gs. | treatment of crude stimulatory gtp-binding protein of adenylyl cyclase (gs) from turkey erythrocyte membranes with hydroxylamine results in twofold enhancement of adenylyl cyclase activity following reconstitution with adenylyl cyclase type v expressed in spodoptera frugiperda cells (sf9) cells. enhancement by hydroxylamine of immunoaffinity purified gs was still 1.5-fold, while that of gs purified according to the multiple-step procedure by northup, j. k., sternweis, p. c., smigel, m. d., schle ... | 1998 | 9799124 |
| nineteen baculovirus open reading frames, including lef-12, support late gene expression. | a set of 18 plasmid subclones of the autographa californica nuclear polyhedrosis virus genome, each containing an identified late expression factor gene (lef), supports expression from a late viral promoter in transient expression assays in the sf-21 cell line derived from spodoptera frugiperda. we have constructed a further set of plasmids in which each lef open reading frame (orf) is controlled by the drosophila melanogaster heat shock protein 70 (hsp70) promoter and epitope tagged. failure of ... | 1998 | 9811761 |
| infection of lepidoptera with a pseudotyped retroviral vector. | studies requiring the introduction and expression of manipulated gene constructs have been technically difficult in non-drosophilid insects. retroviruses can be engineered to be replication defective and to serve as vectors for gene constructs of interest. in this study, pseudotyped momlv(vsv-g) retroviral vectors are shown to successfully infect lepidopteran cells in vitro and in vivo. in spodoptera frugiperda cells in vitro and in manduca sexta in vivo, infection and conversion to proviral dna ... | 1998 | 9818383 |
| recombinant ebola virus nucleoprotein and glycoprotein (gabon 94 strain) provide new tools for the detection of human infections. | after cloning and sequencing the glycoprotein (gp) gene of one of the gabonese strains of ebola virus isolated during the 1994-1996 outbreak, it was shown that the circulating virus was of the zaire subtype. this was confirmed in this study by cloning and sequencing the nucleoprotein (np) gene of this strain. these two structural proteins were also expressed as recombinant proteins and used in elisa tests. np was expressed as a his-tagged fusion protein in escherichia coli and was purified on re ... | 1998 | 9820131 |
| the us5 open reading frame of herpes simplex virus type 1 does encode a glycoprotein (gj). | based on sequence analysis, the protein encoded by the us5 open reading frame (orf) of herpes simplex virus type 1 (hsv-1) was predicted to contain an n-glycosylation site and was given the designation of glycoprotein j (gj). however, the us5 gene product has not been identified and the identity of gj as a glycoprotein has not been confirmed. we have cloned and expressed the dna encoding the complete sequence of the us5 orf, using a baculovirus expression system. western blotting, using polyclon ... | 1998 | 9820842 |
| high-level production of alternatively spliced soluble interleukin-6 receptor in serum of patients with adult t-cell leukaemia/htlv-i-associated myelopathy. | we have previously shown, using human t-cell lymphocytotrophic virus-i (htlv-i)-infected cell lines, that soluble interleukin-6 receptor (sil-6r) is generated through an alternative splicing mechanism. in this study, we examined human sera for the presence of alternatively spliced soluble il-6r (as-sil-6r). we produced a monoclonal antibody (mab) recognizing the unique sequence of as-sil-6r peptide, generated by an altered reading frame. we also made recombinant as-sil-6r protein in spodoptera f ... | 1998 | 9824498 |
| encephalitogenicity of myelin-associated oligodendrocytic basic protein and 2',3'-cyclic nucleotide 3'-phosphodiesterase for balb/c and sjl mice. | in search of new encephalitogenic myelin antigens, the 2',3'-cyclic nucleotide 3'-phosphodiesterase (cnp) and 19 000 mw isoform of myelin-associated oligodendrocytic basic protein (mobp) were obtained as recombinant proteins by the baculovirus expression system in spodoptera frugiperda cells and purified to homogeneity by immobilized metal chelate affinity chromatography (imac). the purified mobp was soluble in water and showed retarded migration on sodium dodecyl sulphate-polyacrylamide gel ele ... | 1998 | 9824501 |
| cellular oxygenation of 12-hydroxyeicosatetraenoic acid and 15-hydroxyeicosatetraenoic acid by 5-lipoxygenase is stimulated by 5-lipoxygenase-activating protein. | it has been proposed that 5-lipoxygenase (5-lo)-activating protein (flap) is an arachidonate transfer protein for leukotriene biosynthesis. using the spodoptera frugiperda (sf9) insect cells, we demonstrate that flap causes a large stimulation (190-fold) of the conversion of 12(s)-hydroxyeicosatetraenoic acid (12(s)-hete) to 5, 12-dihete when co-expressed with 5-lipoxygenase. we also demonstrate that flap can stimulate (2-2.5-fold) the oxygenation of 15(s)-hete by 5-lo to 5,15-dihete. the stimul ... | 1998 | 9830031 |
| induction of t cell response by bluetongue virus core-like particles expressing a t cell epitope of the m1 protein of influenza a virus. | a cd4+ t cell epitope of the influenza virus matrix protein corresponding to the c terminus (qayqkrmgvqmqrfk) was inserted into the vp7 gene of bluetongue virus (btv). the chimeric protein was expressed by a dual recombinant autographa californica polyhedrosis virus (acnpv), which encodes the two inner capsid proteins vp3 and vp7 of btv. when spodoptera frugiperda cells (sf9 cells) were infected with this recombinant btv, core-like particles (clps) were formed as demonstrated by electron microsc ... | 1998 | 9832322 |
| pharmacological properties of homomeric and heteromeric glur1o and glur3o receptors. | homomeric and heteromeric alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionate (ampa) receptor subunits glur1o and glur3o were expressed in spodoptera frugiperda (sf9) insect cells. membranes containing the recombinant receptors showed a doublet of bands of the expected size (99-109 kda) after western immunoblotting which was shifted to a single band upon deglycosylation. in (r,s)-[3h]ampa binding experiments, high expression was seen (bmax = 0.8-3.8 pmol/mg protein) along with high affinity bin ... | 1998 | 9851590 |
| production of recombinant porcine tumor necrosis factor alpha in a baculovirus expression system. | porcine tumor necrosis factor alpha (tnf-alpha) was produced using a baculovirus system in spodoptera frugiperda (sf21ae) cells. cytotoxic activity was detected in supernatant of sonicated sf21ae cells infected with recombinant viruses. the recombinant protein was also demonstrated to be functionally active by its ability to cause apoptosis in wehi 164 cells. three distinct bands of 26, 17 and 14 kda were revealed by western blot analysis using anti-human tnf-alpha antibody. moreover, the anti-h ... | 1998 | 9853300 |
| expression and cellular distribution of baculovirus-expressed bovine herpesvirus 1 (bhv-1) glycoprotein d (gd) sequences. | glycoprotein d (gd) of bovine herpesvirus 1 (bhv-1), a homolog of herpes simplex virus gd, represents a major component of the viral envelope and is a dominant immunogen. to study the antigenic properties of the different regions of gd, we have expressed the full-length gd encoding gene and overlapping fragments spanning various regions of the gd open reading frame in a baculovirus (autographa californica nuclear polyhedrosis virus)--insect cell (spodoptera frugiperda, sf-9) system. maximum leve ... | 1998 | 9856100 |
| substrate specificity and kinetic mechanism of the insect sulfotransferase, retinol dehydratase. | spodoptera frugiperda retinol dehydratase catalyzes the conversion of retinol to the retro-retinoid anhydroretinol. it shares sequence homology with the family of mammalian cytosolic sulfotransferases and provides the first link between sulfotransferases and retinol metabolism. in this study the enzymatic properties of retinol dehydratase were examined using bacterially expressed protein. we show that retinol dehydratase can catalyze the transfer of the sulfonate moiety to small phenolic compoun ... | 1998 | 9857081 |
| characterization of the acidic domain of the ie1 regulatory protein from orgyia pseudotsugata multicapsid nucleopolyhedrovirus. | this study presents a detailed analysis of the acidic n-terminal region of the orgyia pseudotsugata multicapsid nucleopolyhedrovirus (opmnpv) transactivator ie1. the n-terminal region of ie1 is rich in acidic amino acids and has been hypothesized to be an acidic activation domain. removal of the n-terminal 126 amino acids containing the acidic domain of ie1 resulted in complete loss of transactivation activity, indicating that this region is essential for transactivation. the opmnpv acidic domai ... | 1998 | 9875318 |
| lipid composition of spodoptera frugiperda (sf9) and trichoplusia ni (tn) insect cells used for baculovirus infection. | the lipid composition of two different insect cell lines from spodoptera frugiperda (sf9) and trichoplusia ni (tn) which are established cell lines for infection with recombinant baculovirus was analyzed by high-performance liquid chromatography and gas-liquid chromatography. the major phospholipids found were phosphatidylcholine and phosphatidylethanolamine, the major mono-unsaturated fatty acids were oleic acid and palmitoleic acid, the major saturated fatty acid was stearic acid. the choleste ... | 1998 | 9877163 |
| toxicity and receptor binding properties of a bacillus thuringiensis cryic toxin active against both lepidoptera and diptera. | this study describes an investigation into the relationship among toxicity, specificity, and binding of cryic delta-endotoxin from bacillus thuringiensis subspecies aizawai hd-229. in vivo bioassays of larvae using inclusions (aedes aegypti) or solubilized toxin (spodoptera littoralis, spodoptera exigua, spodoptera frugiperda, and manduca sexta) revealed that cryic was toxic to both dipteran and lepidopteran larvae. in vitro saturation binding assays with cryic toxin labeled in vivo with [35s]me ... | 1999 | 9878290 |
| noxious components of venom from the pupa-specific parasitoid pimpla hypochondriaca | venom from the endoparasitoid wasp pimpla hypochondriaca induced a lethal paralysis when injected into the hemocoel of larval, pupal, and adult stages of the tomato moth lacanobia oleracea and was also toxic to the adult stages of the housefly musca domestica and the cockroach blatella germanica. heating the venom to 58 degreesc did not reduce its potency against m. domestica. phenoloxidase activity was detected in diluted venom using dihydroxyphenylalanine as the substrate and occurred independ ... | 1999 | 9878292 |
| development and use of a pcr assay for detection of the reproductive virus in wild populations of helicoverpa zea (lepidoptera: noctuidae) | helicoverpa zea reproductive virus (hzrv) is a nonoccluded bacilliform virus that affects both female and male moths of the corn earworm h. zea. in order to study the biology and host range of hzrv, a bioassay was previously developed to detect the presence of this virus in infected insects. a drawback of this bioassay is that it is time consuming and requires more than a month to complete. here we describe the development of a polymerase chain reaction (pcr) assay for the rapid detection of hzr ... | 1999 | 9878296 |
| isolation of an apoptosis suppressor gene of the spodoptera littoralis nucleopolyhedrovirus. | spodoptera frugiperda sf9 cells infected with mutants of the autographa californica nucleopolyhedrovirus (acmnpv) which lack a functional p35 gene undergo apoptosis, aborting the viral infection. the spodoptera littoralis nucleopolyhedrovirus (slnpv) was able to suppress apoptosis triggered by vdeltap35k/pol+, an acmnpv p35 null mutant. to identify the putative apoptotic suppressor gene of slnpv, overlapping cosmid clones representing the entire slnpv genome were individually cotransfected along ... | 1999 | 9882332 |
| molecular characterization, enzymatic analysis, and purification of murine proprotein convertase-1/3 (pc1/pc3) secreted from recombinant baculovirus-infected insect cells. | a cdna coding for the murine proprotein convertase-1 (mpc1 also known as mpc3 or mspc3) was inserted into the autographa californica nuclear polyhedrosis virus. following infection of spodoptera frugiperda cells, the recombinant n-glycosylated protein is secreted into the cell culture medium from which it can be purified to homogeneity as a fully enzymatically active enzyme. two major secreted molecular forms of mpc1 with apparent molecular weights of 85 and 71 kda, respectively, and a minor one ... | 1998 | 9882569 |
| cloning and expression of a putative glycoprotein gene of channel catfish virus using baculovirus expression system. | the 1035 base-pair (bp) open reading frame 59 (orf59) of channel catfish virus (ccv) codes for an abundant hydrophobic membrane glycoprotein considered to be the major envelope glycoprotein of the virus. the orf59 gene of ccv was cloned into the baculovirus genome by site-specific transposition and expressed in spodoptera frugiperda cells (sf9 cells). expressed protein could be recognized by polyclonal antibody against ccv and appeared to have the same electrophoretic mobility as native protein. ... | 1998 | 9891736 |
| identification and characterization of spodoptera frugiperda furin: a thermostable subtilisin-like endopeptidase. | spodoptera frugiperda (sf9) cells are widely employed for high-level expression of heterologous recombinant genes from baculovirus vectors. using a plasmid library encoding cdna of sf9 cells we have identified here the spodoptera frugiperda analog of the proprotein convertase furin which plays an important role in posttranslational protein processing. spodoptera frugiperda furin (sfurin) is closest related to drosophila melanogasterfurin with which it shares an extended cysteine-rich domain, whe ... | 1998 | 9894811 |
| characterization of the leukotriene b4 receptor in porcine leukocytes. separation and reconstitution with heterotrimeric gtp-binding proteins. | leukotriene b4 (ltb4) is a potent chemoattractant derived from arachidonic acid. when cdnas for ltb4 receptor (blt) were cloned it was found that they belong to a guanine nucleotide-binding regulatory protein (g-protein)-coupled receptor superfamily. however, purification of blt from inflammatory cells and reconstitution with various types of g-proteins have not been successful. in the present study, blt from porcine leukocytes was solubilized, separated from associated g-proteins by ricinus com ... | 1999 | 9914522 |
| aberrant expression and dysfunction of fas antigen in mrl/mpj-lpr/lpr murine ovary. | in lpr mice the insertion of an early transposable element (etn) into intron 2 of the fas gene, which mediates apoptosis, causes the development of massive lymphadenopathy, splenomegaly and autoimmune disease. in the present study we investigated the influence of this mutation on ovarian development of lpr mice. by means of in situ hybridisation, the expression of fas mrna was detected at the same levels in the ovarian cells of mrl/mpj-lpr/lpr (mrl/lpr) mice as in those of mrl/mpj-(+)/+ (mrl/+) ... | 1998 | 9921647 |
| toxicity, binding, and permeability analyses of four bacillus thuringiensis cry1 delta-endotoxins using brush border membrane vesicles of spodoptera exigua and spodoptera frugiperda. | the binding and pore formation properties of four bacillus thuringiensis cry1 toxins were analyzed by using brush border membrane vesicles from spodoptera exigua and spodoptera frugiperda, and the results were compared to the results of toxicity bioassays. cry1fa was highly toxic and cry1ac was nontoxic to s. exigua and s. frugiperda larvae, while cry1ca was highly toxic to s. exigua and weakly toxic to s. frugiperda. in contrast, cry1bb was active against s. frugiperda but only marginally activ ... | 1999 | 9925568 |
| cloning and expression of a novel juvenile hormone-metabolizing epoxide hydrolase during larval-pupal metamorphosis of the cabbage looper, trichoplusia ni. | a full-length cdna encoding for a microsomal juvenile hormone (jh)-metabolizing epoxide hydrolase (tmeh-1) was isolated from a cdna library constructed from fat body of last stadium (wandering) cabbage loopers, trichoplusia ni, at the exact developmental time of maximum jh epoxide hydrolase activity. tmeh-1 was 1887 base pairs in length with a 1389 base pair open reading frame encoding 463 amino acids. amino acid sequence analysis showed that tmeh-1 was most similar to and contained the exact ca ... | 1999 | 9927177 |
| in vivo and in vitro analysis of baculovirus ie-2 mutants. | upon transient expression in cell culture, the ie-2 gene of autographa californica nuclear polyhedrosis virus (acmnpv) displays three functions: trans activation of viral promoters, direct or indirect stimulation of virus origin-specific dna replication, and arrest of the cell cycle. the ability of ie2 to trans stimulate dna replication and coupled late gene expression is observed in a cell line derived from spodoptera frugiperda but not in a cell line derived from trichoplusia ni. this finding ... | 1999 | 9971831 |
| a plant 126-kda phosphatidylinositol 4-kinase with a novel repeat structure. cloning and functional expression in baculovirus-infected insect cells. | phosphatidylinositol metabolism plays a central role in signaling pathways in animals and is also believed to be of importance in signal transduction in higher plants. we report here the molecular cloning of a cdna encoding a previously unidentified 126-kda phosphatidylinositol (pi) 4-kinase (atpi4kbeta) from the higher plant arabidopsis thaliana. the novel protein possesses the conserved domains present in animal and yeast pi 4-kinases, namely a lipid kinase unique domain and a catalytic domain ... | 1999 | 10026194 |
| evaluation of transgenic sweet corn hybrids expressing cryia (b) toxin for resistance to corn earworm and fall armyworm (lepidoptera: noctuidae). | many of the lepidopterous insects which attack sweet corn, zea mays l., are susceptible to insecticidal proteins produced by bacillus thuringiensis ssp. kurstaki (berliner) (btk). transgenic sweet corn expressing a synthetic cry gene for production of a btk-insecticidal protein may provide a more environmentally acceptable means of sweet corn production. eight transgenic sweet corn hybrids containing a synthetic gene for cryia(b) protein production (bt11 event) were evaluated for resistance to t ... | 1999 | 10036986 |
| identification of additional genes that influence baculovirus late gene expression. | we were unable to confirm transient late gene expression using constructs of 18 genes that had been reported to support autographa californica multinucleocapsid nucleopolyhedrovirus (acmnpv) late gene expression when transfected into spodoptera frugiperda cells [lu, a., and miller, l. k. (1995). j. virol. 69, 975-982]. three genes (orf66, orf68, and orf41) were included, all or in part, in the constructs used in that study, but they had not been independently tested. therefore we investigated th ... | 1999 | 10049816 |
| release and preparation of intact and unreduced n-linked oligosaccharides from sf-9 insect cells. | glycosylation, the addition of carbohydrates to a peptide backbone, is the most extensive cotranslational and posttranslational modification made to proteins by eukaryotic cells. the glycosylation profile of a recombinant glycoprotein can significantly affect its biological activity, which is particularly important when being used in human therapeutic applications. therefore, defining glycan structures to ensure consistency of recombinant glycoproteins among different batches is critical. in thi ... | 1999 | 10069429 |
| the biosynthesis and post-translational modification of pbs21 an ookinete-surface protein of plasmodium berghei. | radiolabelled methionine incorporation into synchronised plasmodium berghei gametocytes or ookinete cultures, showed that pbs21 is not synthesised in bloodstage parasites; synthesis was detected within three hours of induction of gametogenesis; synthesis was triggered at gametogenesis, not by fertilisation. we show native pbs21 to be a hydrophobic membrane protein that was insensitive to cleavage by phosphatidylinositol phospholipase c (pi-plc), but sensitive to alkaline hydroxylamine, and parti ... | 1999 | 10080386 |
| o-glycosylation potential of lepidopteran insect cell lines. | the enzyme activities involved in o-glycosylation have been studied in three insect cell lines, spodoptera frugiperda (sf-9), mamestra brassicae (mb) and trichoplusia ni (tn) cultured in two different serum-free media. the structural features of o-glycoproteins in these insect cells were investigated using a panel of lectins and the glycosyltransferase activities involved in o-glycan biosynthesis of insect cells were measured (i.e., udp-galnac:polypeptide n-acetylgalactosaminyltransferase, udp-g ... | 1999 | 10082987 |
| differences in the expression and localization of human melanotransferrin in lepidopteran and dipteran insect cell lines. | the ability of several lepidopteran and dipteran insect cell lines to express human melanotransferrin (p97), a glycosyl phosphatidylinositol (gpi)-anchored, iron-binding sialoglycoprotein, was assessed. spodoptera frugiperda-derived (sf9) cell lines, transformed with the p97 gene under control of a baculovirus immediate-early promoter, were able to constitutively express the protein and correctly attach it to the outer cell membrane via a gpi anchor as demonstrated by pi-plc treatment. in contra ... | 1999 | 10092489 |
| optimising fed-batch production of recombinant proteins using the baculovirus expression vector system. | fed-batch culture can offer significant improvement in recombinant protein production compared to batch culture in the baculovirus expression vector system (bevs), as shown by nguyen et al. (1993) and bedard et al. (1994) among others. however, a thorough analysis of fed-batch culture to determine its limits in improving recombinant protein production over batch culture has yet to be performed. in this work, this issue is addressed by the optimisation of single-addition fed-batch culture. this t ... | 1998 | 10099329 |
| synthesis of rotavirus-like particles in insect cells: comparative and quantitative analysis. | when the three major structural proteins, vp2, vp6, and vp7, of rotavirus are co-expressed in insect cells infected with recombinant baculoviruses, they self-assemble into triple-layered virus-like particles (vlps) that are similar in morphology to native rotavirus. in order to establish the most favorable conditions for the synthesis of rotavirus vlps, we have compared the kinetics of 2/6/7-vlp synthesis in two different insect cell lines: high five cells propagated in excell 405 medium and spo ... | 1998 | 10099441 |
| use of mannosamine for inducing the addition of outer arm n-acetylglucosamine onto n-linked oligosaccharides of recombinant proteins in insect cells. | glycosylation is a cellular process accomplished by a series of sequential enzymatic processing steps through the endoplasmic reticulum and golgi apparatus with vesicle transport between the membranous organelles. the capacity for complex glycosylation is considered to be conferred by cell genetics, while the roles of nongenetic factors in protein processing are often ignored. it was hypothesized that the glycosyltransferase reactions in the insect cell-baculovirus system were limited by the sma ... | 1999 | 10194391 |
| induction of protective immunity against dengue virus type 2: comparison of candidate live attenuated and recombinant vaccines. | dengue (den) viruses (serotypes 1 to 4) are mosquito-borne flaviviruses which cause about fifty million human infections annually and represent an expanding public health problem in the tropics. at present, there are no safe and effective vaccines which induce protective immunity to all four serotypes of den. natural infection or vaccination with native and recombinant proteins may induce an immune response to the surface envelope e-protein which was shown to be protective to super-infection wit ... | 1999 | 10195766 |
| identification of a cell surface protein from crandell feline kidney cells that specifically binds aleutian mink disease parvovirus. | aleutian mink disease parvovirus (adv) is the etiological agent of aleutian disease of mink. the acute disease caused by adv consists of permissive infection of alveolar type ii cells that results in interstitial pneumonitis. the permissive infection is experimentally modeled in vitro by infecting crandell feline kidney (crfk) cells with a tissue culture-adapted isolate of adv, adv-g. adv-g vp2 empty virions expressed in a recombinant baculovirus system were analyzed for the ability to bind to t ... | 1999 | 10196278 |
| parameters influencing human mu opioid receptor over-expression in baculovirus-infected insect cells. | the cdna encoding the human mu opioid receptor (hmor) was cloned in the baculovirus autographa californica (acmnpv) under the control of the polyhedrin promoter. we investigated the influence of different molecular constructions on receptor expression levels: the receptor was fused either to an amino- or a carboxy-terminal histidine tag (hmor-n-his and hmor-c-his respectively), or to the cleavable sequence signal of the baculovirus gp64 glycoprotein (gp-hmor and gp-hmor-c-his). two cell lines, s ... | 1999 | 10201114 |
| molecular cloning and functional expression of o-methyltransferases common to isoquinoline alkaloid and phenylpropanoid biosynthesis. | in cell suspension cultures of the meadow rue thalictrum tuberosum, biosynthesis of the anti-microbial alkaloid berberine can be induced by addition of methyl jasmonate to the culture medium. the activities of the four methyltransferases involved in the formation of berberine from l-tyrosine are increased in response to elicitor addition. partial clones generated by rt-pcr with methyltransferase-specific primers were used as hybridization probes to isolate four cdnas encoding o-methyltransferase ... | 1999 | 10205892 |
| use of the baculovirus system to assemble polyomavirus capsid-like particles with different polyomavirus structural proteins: analysis of the recombinant assembled capsid-like particles. | the genes encoding the structural proteins (vp1, vp2 and vp3) of murine polyomavirus were cloned into the p2bac dual multiple cloning site vector, individually or jointly, and the corresponding proteins were expressed in spodoptera frugiperda (sf9) insect cells by cotransfecting sf9 cells with the constructed vector and the linear dna of autographa californica multiple nuclear polyhedrosis virus (acmnpv). recombinant capsid-like particles could be purified 5 days post-infection from sf9 cells in ... | 1999 | 10211971 |
| in vitro host range of autographa californica nucleopolyhedrovirus recombinants lacking functional p35, iap1 or iap2. | we have examined the host range in different insect cell lines of autographa californica nucleopolyhedrovirus (acmnpv) recombinants lacking p35, iap1 or iap2. these genes encode, or are predicted to encode, anti-apoptotic proteins. abrogation of p35 reduced the ability of acmnpv to replicate in permissive cell lines derived from spodoptera frugiperda insects by inducing apoptosis. in semi-permissive cell lines, such as lymantria dispar and spodoptera littoralis cells, we observed cytopathic effe ... | 1999 | 10211976 |
| the baculovirus antiapoptotic p35 gene also functions via an oxidant-dependent pathway. | cellular imbalance in the levels of antioxidants and reactive oxygen species resulting in apoptosis is directly associated with a number of parasitic infections, aging, and several genetic and multifactorial diseases. the baculovirus acnpv-encoded antiapoptotic p35 gene prevents apoptosis induced by a variety of apoptotic agents in different systems. we demonstrate the ability of the p35 gene to inhibit oxidative stress-induced apoptosis. in vitro cultured spodoptera frugiperda (sf9) insect cell ... | 1999 | 10220380 |
| development of hiv/aids vaccine using chimeric gag-env virus-like particles. | we attempted to develop a candidate hiv/aids vaccine, by using unprocessed hiv-2 gag pr45 precursor protein. we found that a 45 kda unprocessed hiv-2 gag precursor protein (pr45), with a deletion of a portion of the viral protease, assembles as virus-like particles (vlp). we mapped the functional domain of hiv-2 gag vlp formation in order to find the minimum length of gag protein to form vlp. a series of deletion mutants was constructed by sequentially removing the c-terminal region of hiv-2 gag ... | 1999 | 10223338 |
| cell cycle arrest mediated by hepatitis delta antigen. | hepatitis delta antigen (hdag) is the only viral-encoded protein of the hepatitis delta virus (hdv). this protein has been extensively characterized with respect to its biochemical and functional properties. however, the molecular mechanism responsible for persistent hdv infection is not yet clear. previously, we reported that overexpression of hdag protects insect cells from baculovirus-induced cytolysis [hwang, s.b. park, k.-j. and kim, y.s. (1998) biochem. biophys. res. commun. 244, 652-658]. ... | 1999 | 10225424 |
| real-time fluorimetric analysis of gramicidin d- and alamethicin-induced k+ efflux from sf9 and cf1 insect cells. | gramicidin d and alamethicin are pore-forming peptides which exhibit lethal properties against a large spectrum of cells. despite a wealth of experimental data from artificial membranes, the time course and quantitative analysis of the activity of these ionophores are not well described in living cells. in the present study, the newly described fluorescent dye cd-222 was used to monitor extracellular potassium ion concentration and report the effects of these antibiotics on the k+ permeability o ... | 1999 | 10320344 |
| coinfection of spodoptera exigua and spodoptera frugiperda cell lines with the nuclear polyhedrosis viruses of autographa californica and spodoptera exigua. | the autographa californica multinucleocapsid nuclear polyhedrosis virus (acmnpv) has a broad host range among lepidoptera. in contrast, the spodoptera exigua mnpv (semnpv) can replicate efficiently only in s. exigua larvae or s. exigua-derived cell lines. in this study, we examined the coinfection of s. exigua se301 and spodoptera frugiperda iplb-sf21aeii (sf21) cell lines with semnpv and acmnpv recombinant (ac360-501beta-gal) which was constructed for expression of beta-galactosidase under cont ... | 1998 | 10325534 |
| temperature dependent characteristics of a recombinant infectious hematopoietic necrosis virus glycoprotein produced in insect cells. | a recombinant infectious hematopoietic necrosis virus (ihnv) glycoprotein (g protein) was produced in insect cells using a baculovirus vector (autographa californica nuclear polyhedrosis virus). characteristics of this protein were evaluated in relation to native viral g protein. a full-length (1.6 kb) cdna copy of the glycoprotein gene of ihnv was inserted into the baculovirus vector under control of the polyhedrin promoter. high levels of g protein (approximately 0.5 microgram/1 x 10(5) cells) ... | 1999 | 10349547 |
| immunogenicity of a recombinant infectious hematopoietic necrosis virus glycoprotein produced in insect cells. | a recombinant infectious hematopoietic necrosis virus (ihnv) glycoprotein (g protein), produced in spodoptera frugiperda (sf9) cells following infection with a baculovirus vector containing the full-length (1.6 kb) glycoprotein gene, provided very limited protection in rainbow trout oncorhynchus mykiss challenged with ihnv. fish were injected intraperitoneally (i.p.) with sf9 cells grown at 20 degrees c (recglow) or 27 degrees c (recghigh) expressing the glycoprotein gene. various antigen (ag) p ... | 1999 | 10349553 |
| replication of spodoptera exigua nucleopolyhedrovirus in permissive and non-permissive lepidopteran cell lines. | the spodoptera exigua multinucleocapsid nucleopolyhedrovirus (semnpv) was inoculated to eight lepidopteran cell lines derived from spodoptera exigua (se301), spodoptera frugiperda (sf21aeii), spodoptera littoralis (cls-79), spodoptera litura (spli-221), pseudaletia separata (lese-11), trichoplusia ni (hi-5), plutella xylostella (pxl/c) and bombyx mori (bmn4). the productive infection of semnpv was observed only in se301 cells. however, a dot-blot hybridization analysis revealed that semnpv dna r ... | 1998 | 10358729 |
| generation of a new protein purification matrix by loading ceramic hydroxyapatite with metal ions--demonstration with poly-histidine tagged green fluorescent protein. | the gene encoding the green fluorescent protein (gfp) from the jellyfish aequorea victoria, was inserted under transcriptional control of the polyhedrin promoter of the autographa californica nuclear polyhedrosis virus and expressed in the spodoptera frugiperda insect cell line sf9 during viral infection. the baculovirus transfervector pbluebachisb was used for constructing the recombinant baculovirus, so that the green fluorescent protein could be tagged with a poly-histidine tail. this fusion ... | 1999 | 10361722 |
| host cell receptor binding by baculovirus gp64 and kinetics of virion entry. | gp64 is the major envelope glycoprotein from budded virions of the baculoviruses autographa californica multicapsid nucleopolyhedrovirus (acmnpv) and orgyia pseudotsugata multicapsid nucleopolyhedrovirus (opmnpv). to examine the potential role of gp64 as a viral attachment protein in host cell receptor binding, we generated, overexpressed, and characterized a soluble form of the opmnpv gp64 protein, gp64solop. assays for trimerization, sensitivity to proteinase k, and reduction by dithiothreitol ... | 1999 | 10366584 |
| circular configuration of the genome of ascoviruses. | a circular configuration of genomic dna was observed in ascoviruses isolated from two species of insects of the family noctuidae [fall armyworm (spodoptera frugiperda) and cotton bollworm (helicoverpa zea)] using restriction endonuclease (ren) digestion, conventional gel electrophoresis, pulsed-field gel electrophoresis and southern hybridization analysis. this circular configuration of ascovirus genomic dna was established based on the difference between linear and circular dna in the numbers o ... | 1999 | 10374973 |
| role of murine cytochrome p-450 2f2 in metabolic activation of naphthalene and metabolism of other xenobiotics. | despite their substantially lower levels relative to hepatic tissue, pulmonary cytochrome p-450 (cyp) monooxygenases play an important role in the metabolic activation of substrates that cause lung injury. the target- and species-selective toxicity of a number of pulmonary toxicants has been attributed to the presence and distribution of activating enzymes with high kcat in target airways of susceptible species. however, experimental demonstration of these concepts and quantitative assessment of ... | 1999 | 10381788 |
| mitochondrial dna acts as potential promoter of the baculovirus rna polymerase. | we have examined whether mitochondrial dna could act as target of the rna polymerase encoded by the baculovirus autographa californica multicapsid nuclear polyhedrosis virus, because the baculovirus late promoters and the control region of host mitochondrial dna show a high degree of sequence similarity. in vitro transcription using mitochondrial dna from spodoptera frugiperda cells and nuclear extracts prepared from baculovirus infected cells demonstrates that mitochondrial dna is recognized by ... | 1999 | 10384964 |
| baculovirus expression, purification and evaluation of recombinant pneumococcal surface adhesin a of streptococcus pneumoniae. | pneumococcal surface adhesin a (psaa), with a molecular mass of approximately 37 kd by sds-page, is a common surface protein expressed by all 90 serotypes of streptococcus pneumoniae. s. pneumoniae serotype 6b genomic dna was amplified to generate a dna fragment carrying the full-length psaa sequence and was cloned into a baculovirus expression system. we expressed either cell-associated or cell-free nonfusion psaa polypeptides using two insect cell lines, spodoptera frugiperda (sf9) and trichop ... | 1999 | 10394131 |
| characterization of three distinct cdna clones encoding cysteine proteinases from maize (zea mays l.) callus. | in previous work, a 33 kda cysteine proteinase was found in callus initiated from maize (zea mays l.) resistant to fall armyworm feeding. a callus cdna library from the maize inbred mp708 was screened with oligonucleotides derived from the n-terminal amino acid sequence of the 33 kda proteinase and several cdna clones were isolated and sequenced. a cdna clone encoding the 33 kda cysteine proteinase, mir1, was identified. two additional clones, mir2 and mir3, encoding putative cysteine proteinase ... | 1999 | 10394950 |
| recombinant protein synthesis in trichoplusia ni bti-tn-5b1-4 insect cell aggregates. | the trichoplusia ni bti-tn-5b1-4 (tn-5b1-4) insect cell line has received considerable attention as a host for the baculovirus expression vector system. in the present study, suspension cultures were used to compare tn-5b1-4 cell aggregates and cells selected to grow predominantly as individual cells. no significant difference was found between cell aggregates and cells growing predominantly individually in regard to cell growth rate, glucose consumption and lactate accumulation, and specific re ... | 1999 | 10397817 |
| chimeric infectious bursal disease virus-like particles expressed in insect cells and purified by immobilized metal affinity chromatography. | chimeric virus-like particles (vlps) of infectious bursal disease virus (ibdv) were produced by coinfecting spodoptera frugiperda (sf-9) insect cells with two recombinant baculoviruses, vibd-7 and vedlh-22. vibd-7 encodes vp2, vp3, and vp4 of the ibdv structural proteins. vedlh-22 encodes vp2 with five histidine residues at the carboxy-terminus (vp2h). coinfection produced hybrid vlps composed of vp2, vp2h, and vp3. the additional histidine residues on vp2h enabled the efficient purification of ... | 1999 | 10397829 |
| characterization of bimodal cell death of insect cells in a rotating-wall vessel and shaker flask | in previous publications, we reported the benefits of a high-aspect rotating-wall vessel (harv) over conventional bioreactors for insect-cell cultivation in terms of reduced medium requirements and enhanced longevity. to more fully understand the effects that harv cultivation has on longevity, the present study characterizes the mode and kinetics of spodoptera frugiperda cell death in this quiescent environment relative to a shaker-flask control. data from flow cytometry and fluorescence microsc ... | 1999 | 10397835 |
| expression, purification, and characterization of the cytoplasmic domain of the human igf-1 receptor using a baculovirus expression system. | the cytoplasmatic domain of the beta-subunit of the human igf-1 receptor (residues 929-1337) has been overexpressed in insect cells using the baculovirus expression system. synthesis of the soluble protein (igfk, m(r) 46 kda) in spodoptera frugiperda (sf9) cells was detected 24 h after infection and maximal accumulation was achieved 40-48 h postinfection. rapid purification to near homogeneity (>/=95% pure protein) was accomplished by sequential chromatography on resource-q and phenyl-sepharose ... | 1999 | 10403833 |
| [vp4 protein of simian rotavirus strain sa11 expressed by a baculovirus recombinant]. | the complementary dna copies of the gene 4 were prepared by reverse transcription and polymerase chain reaction (rt-pcr) from genomic rna of simian rotavirus strain sa11. a complete vp4 gene was been inserted into a baculovirus intermediate vector pvl-1393 which was under control of the polyhedrin promoter. the outer capsid protein vp4 which was a major neutralization antigen of rotavirus, was expressed in high yield in spodoptera frugiperda cell line. the vp4 protein expressed was recognized wi ... | 1997 | 10453552 |
| structure-function analysis of human cytochrome p-450 2b6 using a novel substrate, site-directed mutagenesis, and molecular modeling. | the structural basis for functional differences between human cytochrome p-450 2b6 and rat 2b1 was investigated. an amino acid sequence alignment predicted the location of 2b6 substrate recognition site (srs) residues. ten residues within these srss unique to 2b6 compared with 2b1, 2b4, and 2b11 were chosen for mutagenesis. two additional sites that differ between 2b6 and 2b1 and are known to have a role in 2b1 substrate specificity were also mutated. the 2b6 mutants were expressed in spodoptera ... | 1999 | 10454488 |
| characterization of a bioassay for detection of recombinant and native ovine interleukin-5. | in order to analyse th2-type immune responses in sheep by the assay of interleukin (il)-5 in biological fluids, the ovine il-5 gene was cloned and expressed in spodoptera frugiperda sf9 insect cells using the baculovirus expression vector system. the recombinant product was purified as bac-ov-il-5 from the supernatant fluid. the ovine il-5 was biologically active in a bioassay using il-5-dependent baf cells, which have been used previously to specifically detect human il-5. the specificity of ba ... | 1999 | 10457200 |
| in vitro analysis of venom from the wasp nasonia vitripennis: susceptibility of different cell lines and venom-induced changes in plasma membrane permeability. | the lethal effects of crude venom prepared from the ectoparasitic wasp nasonia vitripennis were examined with cultured cells from six insect and two vertebrate species. venom caused cells from sarcophaga peregrina (nih sape4), drosophila melanogaster (crl 1963), trichoplusia ni (tn-368 and bti-tn-5b1-4), spodoptera frugiperda (sf-21ae), and lymantria dispar (ipl-ldfbc1) to round up, swell, and eventually die. despite similar sensitivities and overlapping lc50 values [0.0004-0.0015 venom reservoi ... | 1999 | 10475265 |
| in vivo and in vitro phosphorylation of rice dwarf phytoreovirus pns12 cytoplasmic nonstructural protein. | in vivo and in vitro phosphorylation and intracellular location of rice dwarf phytoreovirus pns12, which is encoded by one of the twelve dsrna genome segments, s12, and comprises 312 amino acids, was investigated. when [32p]phosphoric acid was incorporated into rdv-infected leafhopper cultured cells, labelled pns12 was immunoprecipitated from the cells by a monospecific anti-pns12 polyclonal antibody. recombinant pns12 was purified from spodoptera frugiperda cells infected with acrs12, a baculov ... | 1999 | 10481743 |
| alteration of glucose consumption kinetics with progression of baculovirus infection in spodoptera frugiperda cells. | we have used the initial-rate approach to characterize changes in the glucose consumption kinetics of baculovirus-infected spodoptera frugiperda clone 9 (sf9) cells with the progression of the infection process. the specific glucose consumption rate (qg) of cultured baculovirus-infected sf9 cells was measured at 4, 8, 12, 16, and 24 h postinfection (h.p.i.) in media containing 4-35 mm glucose. higher medium glucose concentrations resulted in higher final extracellular virus and recombinant beta- ... | 1999 | 10488553 |
| expression of a glycosylphosphatidylinositol-linked manduca sexta aminopeptidase n in insect cells. | aminopeptidase n (apn; ec 3.4.11.2) is an exopeptidase that is attached to cell membranes by a hydrophobic amino-terminal stalk in vertebrates or a glycosylphosphatidylinositol (gpi) anchor in insects. in this study, we report the cloning, expression, and characterization of an aminopeptidase n from manduca sexta midgut. the full-length aminopeptidase n cdna (apn1a) encodes a 995-amino-acid protein. the predicted amino acid sequence differs by 8 amino acids from m. sexta apn1. these different am ... | 1999 | 10497076 |
| expression of the fusion protein recombinant human granulocyte-macrophage colony stimulating factor and leukemia inhibitory factor in a baculovirus vector system. | a fusion gene coding human granulocyte-macrophage colony stimulating factor (gm-csf) and leukemia inhibitory factor (lif) cdnas was inserted into the transfer vector psxivvi+ x3 with the control of syn and xiv promoters. the sf9 cells (spodoptera frugiperda) were co-transfected with the recombinant plasmid and tnnpv dna (trichoplusia ni nuclear polyhedrosis virus dna). the fusion protein recombinant human granulocyte-macrophage colony stimulating factor (gm-csf) and leukemia inhibitory factor (l ... | 1999 | 10510487 |
| expression, purification and characterization of peanut (arachis hypogaea) agglutinin (pna) from baculovirus infected insect cells. | peanut (arachis hypogaea) seed lectin, pna is widely used to identify tumor specific antigen (t-antigen), galbeta1-3galnac on the eukaryotic cell surface. the functional amino acid coding region of a cdna clone, pbsh-pn was pcr amplified and cloned downstream of the polyhedrin promoter in the autographa californica nucleopolyhedrovirus (acnpv) based transfer vector pvl1393. co-transfection of spodoptera frugiperda cells (sf9) with the transfer vector, pacpna and acrp6 (a recombinant acnpv having ... | 1999 | 10513900 |
| molecular characterization of genes in the gp41 region of baculoviruses and phylogenetic analysis based upon gp41 and polyhedrin genes. | a newly sequenced anticarsia gemmatalis multiple nucleopolyhedrovirus (agmnpv) gp41 gene was used to reconstruct the phylogeny for gp41 by comparison with autographa californica mnpv, bombyx mori mnpv, helicoverpa zea single nucleopolyhedrovirus (snpv), lymantria dispar mnpv, orgyia pseudotsugata mnpv and spodoptera frugiperda mnpv. the 3.5 kb fragment of the agmnpv gp41 region not only contained the gp41 gene but also three other open reading frames that had significant homology with the very l ... | 1999 | 10518714 |
| detection of baculovirus-infected insect cells by flow cytometric side-scatter analyses. | the baculovirus expression vector system (bevs), utilizing the autographa californica nuclear polyhedrosis virus (acnpv), has turned out to be an attractive alternative for high-level expression (<600 mg/l) of recombinant proteins. however, there is a shortage of reliable methods for monitoring the infection process in situations where marker proteins cannot be used. | 1999 | 10520205 |
| role of a potent inhibitory monoclonal antibody to cytochrome p-450 3a4 in assessment of human drug metabolism. | cytochrome p-450 (cyp) 3a4 is an inordinately important cyp enzyme that catalyzes the metabolism of a vast array of clinically used drugs. microsomal proteins of spodoptera frugiperda (sf21) insect cells infected with recombinant baculoviruses encoding cyp3a4 cdna were used to immunize mice and to develop a monoclonal antibody (mab(3a4a)) specific to cyp3a4 through the use of hybridoma technology. the mab is both a potent inhibitor and a strong binder of cyp3a4. one and 5 microl (0.5 and 2.5 mic ... | 1999 | 10525096 |
| a noda-like virus isolated from the sweetpotato pest spodoptera eridania (cramer) (lep.; noctuidae) | a small isometric virus has been isolated from larvae of the sweetpotato pest spodoptera eridania (cramer) collected near pariacoto, ancash province, peru. it is designated the pariacoto virus (pav). in addition to its high pathogenicity on its natural host spodoptera eridania, pav was found to replicate in spodoptera ochrea (hampson) larvae but not in spodoptera frugiperda (smith) larvae. the size of the viral particle was estimated to be about 30 nm in diameter. polyacrylamide gel electrophore ... | 1999 | 10534414 |
| characterization of a nucleopolyhedrovirus from the black cutworm, agrotis ipsilon (lepidoptera: noctuidae) | the black cutworm, agrotis ipsilon (hufnagel) (lepidoptera: noctuidae), is a serious localized pest of vegetable and field crops. we have characterized a newly discovered baculovirus, the agrotis ipsilon multicapsid nucleopolyhedrovirus (agipmnpv), that was isolated from a. ipsilon in illinois. restriction enzyme fragment profiles of agipmnpv dna were distinct from those of previously described nucleopolyhedroviruses. electron microscopy of agipmnpv-infected tissues indicated that nucleocapsids ... | 1999 | 10534417 |
| the cadherin-like protein is essential to specificity determination and cytotoxic action of the bacillus thuringiensis insecticidal cryiaa toxin. | the bacillus thuringiensis cryiaa toxin binds a cadherin-like protein (btr175) on the brush-border membranes of the bombyx mori midgut columnar cells, which are the targets. by introducing the btr175 gene with a baculovirus, spodoptera frugiperda sf9 cells expressed btr175 protein on the cell membrane and became susceptible to the cryiaa toxin. the toxin bound the cadherin repeat adjacent to the membrane and made a pore that passed inorganic ions, causing the cell to swell and burst. this was no ... | 1999 | 10544269 |
| biological functions of recombinant bovine interleukin 6 expressed in a baculovirus system. | the cdna encoding bovine interleukin 6 (il-6) was obtained from messenger rna extracted from lipopolysaccharide-stimulated bovine kupffer cells by the reverse transcription polymerase chain reaction (rt/pcr), and cloned into the baculovirus vector pvl 1392. insect cells (sf21ae derived from spodoptera frugiperda) infected with the recombinant baculovirus secreted a large amount of 23.7 kd protein into the culture medium. this protein was capable of causing increased haptoglobin production and de ... | 1999 | 10547274 |
| type 3 inositol trisphosphate receptors in rinm5f cells are biphasically regulated by cytosolic ca2+ and mediate quantal ca2+ mobilization. | there are three subtypes of mammalian ins(1,4,5)p(3) (insp(3)) receptor, each of which forms an intracellular ca(2+) channel. biphasic regulation of insp(3) receptors by cytosolic ca(2+) is well documented in cells expressing predominantly type 1 or type 2 insp(3) receptors and might contribute to the regenerative recruitment of ca(2+) release events and to limiting their duration in intact cells. the properties of type 3 receptors are less clear. bilayer recording from insp(3) receptors of rin- ... | 1999 | 10548533 |
| physical maps and virulence of anticarsia gemmatalis nucleopolyhedrovirus genomic variants. | seventeen plaque purified isolates of two viral preparations of anticarsia gemmatalis multiple nucleopolyhedrovirus (agmnpv), were analyzed in terms of the genomic changes after digestion of their dnas with hindiii and psti restriction enzymes. the 1979 agmnpv wild type preparation (agmnpv-'79) resulted in six different variants and the 1985 viral commercial preparation (agmnpv-'85), in eleven. the genomic variation of all the isolates was mapped showing that those from 1985 presented more heter ... | 1999 | 10550671 |
| large-scale expression and purification of the human vitamin d receptor and its ligand-binding domain for structural studies. | we have expressed recombinant human vitamin d receptor and its ligand-binding domain in spodoptera frugiperda (sf9) insect cells with a 30-litre bioreactor. both proteins were purified to apparent homogeneity with yields of 0.5-3.5 mg/l. gel-filtration analyses indicated that the purified human vitamin d receptor and its ligand-binding domain were present as monomers in solution. the purified vitamin d receptor and its ligand-binding domain were demonstrated to bind 1alpha,25-dihydroxyvitamin d( ... | 1999 | 10567209 |
| agonist selective regulation of g proteins by cannabinoid cb(1) and cb(2) receptors. | we have examined the ligand regulation and g protein selectivity of the human cannabinoid cb(1) and cb(2) receptors by an in situ reconstitution technique directly measuring g protein activation. membranes from spodoptera frugiperda cells expressing cb(1) and cb(2) receptors were chaotrope extracted to denature endogenous gtp-binding proteins. the ability of the receptors to catalyze the gdp-gtp exchange of each g protein was then examined with purified bovine brain g(i) and g(o). activation of ... | 1999 | 10570066 |
| limonoids from swietenia macrophylla and s. aubrevilleana. | an investigation of the seeds of swietenia macrophylla and s. aubrevilleana (meliaceae) is reported. three new compounds, augustineolide (1) and 3beta,6-dihydroxydihydrocarapin (2) from s. macrophylla and 6-acetoxyhumilinolide c (3) from s. aubrevilleana were isolated and characterized along with fifteen known compounds. four of the compounds were subjected to an antifeedant bioassay on the final instar larvae of spodoptera frugiperda. the antifeedant activity was comparable to that of bicyclono ... | 1999 | 10579863 |
| selection of a nucleopolyhedrovirus for control of spodoptera frugiperda (lepidoptera: noctuidae): structural, genetic, and biological comparison of four isolates from the americas. | spodoptera frugiperda (j. e. smith) (lepidoptera: noctuidae) is the principal pest of maize in tropical and subtropical regions of the americas. larvae of this species are susceptible to a nucleopolyhedrovirus (npv) which has attracted interest as a potential biocontrol agent. four strains of npv isolated from infected s. frugiperda larvae in the united states, nicaragua, and argentina were subjected to a structural, genetic, and biological comparison to select a candidate isolate for use in bio ... | 1999 | 10582046 |
| constitutive secretion of an endogenous insulin-like peptide binding protein with high affinity for insulin in spodoptera frugiperda (sf9) cell cultures. | serum-free medium from batch cultures of sf9 insect cells was examined for the occurrence of proteins related to the insulin-like growth factor family. we found that the sf9 cell line constitutively produced and secreted a soluble protein with a mw of 27 kda that exerted specific binding to human insulin-like growth factor-i (igf-i) and -ii. moreover, the secreted protein bound human insulin and human proinsulin with higher affinity than igf-i and -ii. the order of affinity to the insulin peptid ... | 1999 | 10600480 |