| 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 |
| expression of a human, neutralizing monoclonal antibody specific to puumala virus g2-protein in stably-transformed insect cells. | we cloned the heavy- and light-chain antibody genes of a human x (humanxmouse) trioma secreting a neutralizing, igg monoclonal antibody to the g2-protein of puumala virus. the antibody genes were inserted separately into plasmid transfer vector piei-4 such that the genes were under control of the baculovirus immediate early gene promoter, iei. trichoplusia ni (tn) cells were co-transfected with these constructs and a selection plasmid containing a neomycin-resistance gene. cloned transformants e ... | 2000 | 11121551 |
| effect of light energy on alkali-released virions from anagrapha falcifera nucleopolyhedrovirus. | we compared the insecticidal activities of occluded and nonoccluded afmnpv baculovirus obtained by dissolving the occlusion bodies (ob) with sodium carbonate. droplet feeding and cotton leaf feeding bioassay techniques were used to determine the dose response against neonate trichoplusia ni (hübner) and loss of insecticidal activity when the virus was exposed to simulated sunlight from a xenon light source. using droplet bioassays to determine a dose response, nonoccluded virus (nov) was 20 time ... | 2000 | 11023735 |
| effects of incubation temperature on the dose-survival time relationship of trichoplusia ni larvae infected with autographa californica nucleopolyhedrovirus. | the interaction between virus multiplication and host development was studied by determining the survival time of trichoplusia ni larvae inoculated with a wide range of doses of autographa californica nucleopolyhedrovirus and incubated at five different temperatures spanning the biologically relevant range. the results support earlier findings that the course of baculovirus infection follows the mathematical description of the birth-death model. both in vivo rate of virus increase and larval gro ... | 2000 | 11023746 |
| 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 |
| expression, purification, and characterization of the human receptor activator of nf-kappab ligand (rankl) extracellular domain. | receptor activator of nf-kappab ligand (rankl) is a type ii transmembrane protein found on osteoblasts which functions as a major determinant of osteoclast differentiation and activation. rankl mediates bone homeostasis through binding to the cognate ligand on osteoclasts, rank, and a soluble decoy receptor, osteoprotegerin (opg). we designed a construct encoding the extracellular domain of human rankl that conformed to reports of native processing. to encourage folding and posttranslational mod ... | 2000 | 11035950 |
| identification, cloning, and characterization of a major cat flea salivary allergen (cte f 1). | an 18 kda protein isolated from saliva of the cat flea, ctenocephalides felis, elicits a positive intradermal skin test (idst) in 100 and 80% of experimental and clinical flea allergic dogs, respectively. using solid-phase enzyme-linked immuno assay (elisa), this protein detected ige in 100 and 80% of experimental and clinical flea allergic dogs, respectively. a cdna (pfsi) encoding a full-length cte f 1 protein was isolated from a c. felis salivary gland cdna library, using a combination of pcr ... | 2000 | 11074254 |
| replication of the gonad-specific virus hz-2v in ld652y cells mimics replication in vivo. | a newly discovered, nonoccluded insect virus, known as gonad-specific virus or hz-2v, was found to replicate differently in two insect cell lines derived from ovarian tissues (tn-368 cells from trichoplusia ni and ld652y from lymantria dispar). differences between these two cell lines were observed in virus plaque forming ability, rate of viral dna replication, time course of infectious virus production, and the mechanism of virus release from infected cells. replication of hz-2v in ld652y cells ... | 2001 | 11161993 |
| 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 |
| trichoplusia ni gloverin, an inducible immune gene encoding an antibacterial insect protein. | by using differential display pcr, we obtained a cdna clone encoding a gloverin homologue from the cabbage looper, trichoplusia ni. the expression of the gene was induced by bacterial infections. the gene codes for a 174 amino acid residue protein, including a signal sequence and a prosegment. the deduced mature protein is 14 kda and shows 58% and 49% identity to p2 from helicoverpa armigera and to hyalophora gloveri gloverin, respectively. the protein was detected in hemolymph and hemocytes fro ... | 2002 | 12044496 |
| unique disulfide bond structures found in st8sia iv polysialyltransferase are required for its activity. | ncam polysialylation plays a critical role in neuronal development and regeneration. polysialylation of the neural cell adhesion molecule (ncam) is catalyzed by two polysialyltransferases, st8sia ii (stx) and st8sia iv (pst), which contain sialylmotifs l and s conserved in all members of the sialyltransferases. the members of the st8sia gene family, including st8sia ii and st8sia iv are unique in having three cysteines in sialylmotif l, one cysteine in sialylmotif s, and one cysteine at the cooh ... | 2001 | 11279095 |
| 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 |
| comparative pathogenesis of helicoverpa zea s nucleopolyhedrovirus in noctuid larvae. | we used a recombinant of helicoverpa zea s nucleopolyhedrovirus containing the hsp70/lacz reporter cassette (hzsnpv-hsp70/lacz) to quantify mortality relationships and to elucidate early pathogenesis in two permissive hosts, heliothis virescens and helicoverpa zea, and one semi-permissive host, trichoplusia ni. fourth instar t. ni were highly resistant to fatal infection both by oral injection of occlusions and by intrahaemocoelic injection of budded virus, indicating the presence of both midgut ... | 2001 | 11413390 |
| metabolic activation of polycyclic aromatic hydrocarbons and other procarcinogens by cytochromes p450 1a1 and p450 1b1 allelic variants and other human cytochromes p450 in salmonella typhimurium nm2009. | a variety of polycyclic aromatic hydrocarbons and their dihydrodiol derivatives, arylamines, heterocyclic amines, and nitroarenes, were incubated with cdna-based recombinant (escherichia coli or trichoplusia ni) systems expressing different forms of human cytochrome p450 (p450 or cyp) and nadph-p450 reductase using salmonella typhimurium tester strain nm2009, and the resultant dna damage caused by the reactive metabolites was detected by measuring expression of umu gene in the cells. recombinant ... | 2001 | 11502724 |
| both lymantria dispar nucleopolyhedrovirus enhancin genes contribute to viral potency. | enhancins are a group of proteins first identified in granuloviruses (gv) that have the ability to enhance nuclear polyhedrosis virus potency. we had previously identified an enhancin gene (e1) in the lymantria dispar multinucleocapsid nucleopolyhedrovirus (ldmnpv) (d. s. bischoff and j. m. slavicek, j. virol. 71:8133-8140, 1997). inactivation of the e1 gene product within the viral genome lowered viral potency by an average of 2.9-fold. a second enhancin gene (e2) was identified when the entire ... | 2001 | 11507209 |
| n-glycan patterns of human transferrin produced in trichoplusia ni insect cells: effects of mammalian galactosyltransferase. | the n-glycans of human serum transferrin produced in trichopulsia ni cells were analyzed to examine n-linked oligosaccharide processing in insect cells. metabolic radiolabeling of the intra- and extracellular protein fractions revealed the presence of multiple transferrin glycoforms with molecular weights lower than that observed for native human transferrin. consequently, the n-glycan structures of transferrin in the culture medium were determined using three-dimensional high performance liquid ... | 2000 | 10929010 |
| cloning, partial purification and in vivo developmental profile of expression of the juvenile hormone epoxide hydrolase of ctenocephalides felis. | cdnas encoding two different epoxide hydrolases (ncfeh1 and ncfeh2) were cloned from a cdna library prepared from the wandering larval stage of the cat flea, ctenocephalides felis. predicted translations of the open reading frames indicated the clones encoded proteins of 464 (cfeh1) and 465 (cfeh2) amino acids. these proteins have a predicted molecular weight of 53 kda and a putative 22 amino acid n-terminal hydrophobic membrane anchor. the amino acid sequences are 77% identical, and both are ho ... | 2002 | 12125060 |
| establishment and characterization of insect cell lines from 10 lepidopteran species. | cell lines from selected lepidopteran species were established for the overall purpose of use in baculovirus production. a total of 36 new cell lines from 10 lepidopteran species were generated, including cell lines from a pyralid, the european corn borer, ostrinia nubilalis, a plutellid, the diamondback moth, plutella xylostella, as well as eight noctuids: the black cutworm, agrotis ipsilon, the celery looper, anagrapha falcifera, the velvetbean caterpillar, anticarsia gemmatalis, the corn earw ... | 2001 | 11515970 |
| life table parameters, reproductive rate, intrinsic rate of increase, and estimated cost of rearing podisus maculiventris (heteroptera: pentatomidae) on an artificial diet. | the impact of an insect-free artificial diet provided at nymphal and/or adult stage upon the developmental rate, life table parameters, and fertility table parameters was examined for podisus maculiventris (say). this study showed that when fed an insect-free artificial diet during both the nymphal and adult stage, developmental time was prolonged, preoviposition period was extended, and reproductive rate (r0) and intrinsic rate of increase (r) were significantly lower than when fed larval insec ... | 2001 | 11777035 |
| sex differences in the metabolism of (+)- and (-)-limonene enantiomers to carveol and perillyl alcohol derivatives by cytochrome p450 enzymes in rat liver microsomes. | (+)-limonene is reported to cause nephropathy in male rats, but not in female rats and other species of animals including mice, rabbits, guinea pigs, and dogs. male rats contain high levels of alpha2u-globulin in kidneys, and it has been shown that limonene and/or its metabolites are able to bind noncovalently to alpha2u-globulin, resulting in an accumulation of protein droplets in the renal tubules. in this study, we investigated whether (+)- and (-)-limonene enantiomers are differentially meta ... | 2002 | 11800592 |
| effects of phytic acid and xanthotoxin on growth and detoxification in caterpillars. | phytic acid is abundant in the fruits and seeds of many plants and is found in foliage to a lesser extent. among its several properties, phytic acid is a potent chelator of essential minerals and proteins; thus, the possibility exists that heme-based enzymes such as cytochrome p450 monooxygenases in herbivores are detrimentally affected by phytic acid via chelation of dietary iron. mortality, growth performance, and p450-mediated metabolism of xanthotoxin, a plant allelochemical, were examined i ... | 2001 | 11545369 |
| plasmodium falciparum: glycosylation status of plasmodium falciparum circumsporozoite protein expressed in the baculovirus system. | we expressed the main surface antigen of plasmodium falciparum sporozoites, the circumsporozoite protein (csp), in high five (trichoplusia ni) insect cells using the baculovirus system. significant amounts of the recombinant protein could be obtained, as judged by sds-page, western blot, and immunofluorescence analysis. the cellular localization for recombinant csp was determined by immunofluorescence. the high fluorescence signal of the permeabilized cells, relative to that of fixed nonpermeabi ... | 2002 | 12243739 |
| partial redistribution of the autographa californica nucleopolyhedrovirus chitinase in virus-infected cells accompanies mutation of the carboxy-terminal kdel er-retention motif. | during virus infection of insect cells, the autographa californica nucleopolyhedrovirus chitinase is localized primarily within the endoplasmic reticulum (er), which is consistent with the presence of a carboxy-terminal er retention motif (kdel). release of chitinase into the extracellular medium appears to be concomitant with terminal cell lysis, rather than by active secretion. in this study, we have shown that mutation of the kdel motif induces a partial redistribution of the chitinase at bot ... | 2002 | 11842263 |
| identification, sequence analysis, and phylogeny of the immediate early gene 1 of the trichoplusia ni single nucleocapsid polyhedrosis virus. | substantial research has been conducted on the immediate early i (ie-1) genes from the prototype baculovirus auographa californica multicapsid nuclear polyhedrosis virus (acmnpv) and the orgyia pseudotsugata multicapsid nuclear polyhedrosis virus (opmnpv). in both cases ie-1 gene products have been implicated in transcriptional activation and repression. in this study an ie-1 homolog was identified from trichoplusia ni single nucleocapsid polyhedrosis virus (tnisnpv). nucleotide sequence analysi ... | 2001 | 11556402 |
| antimicrobial activity of exocrine glandular secretion of chrysomela larvae. | the exocrine glandular secretions of leaf beetle larvae of the taxon chrysomela are well-known defensive devices used against some generalist predators. salicylaldehyde is the major repellent component of secretions emitted by larvae of chrysomela vigintipunctata and c. lapponica, which feed on salicin-rich salicaceae. in this study, we examined whether salicylaldehyde is also active against the entomopathogenic fungus metarhizium anisopliae. the germination and growth of this fungus was strongl ... | 2002 | 11925070 |
| localization of cctbeta in rat brain and overexpression in insect cells. | to study the localization of ctp: phosphocholine cytidylyltransferase beta isoform (cctbeta) in rat brain, its expression in insect cells and enzymatic properties. | 2002 | 11931693 |
| metabolism of (+)- and (-)-limonenes to respective carveols and perillyl alcohols by cyp2c9 and cyp2c19 in human liver microsomes. | limonene, a monocyclic monoterpene, is present in orange peel and other plants and has been shown to have chemopreventive activities. (+)- and (-)-limonene enantiomers were incubated with human liver microsomes and the oxidative metabolites thus formed were analyzed using gas chromatography-mass spectrometry. two kinds of metabolites, (+)- and (-)-trans-carveol (a product by 6-hydroxylation) and (+)- and (-)-perillyl alcohol (a product by 7-hydroxylation), were identified, and the latter metabol ... | 2002 | 11950794 |
| molecular cloning and induction of nuclear receptors from insect cell lines. | fragments of ecr and usp were cloned from two insect cell lines, sf21 and high five cells (derived respectively from spodoptera frugiperda and trichoplusia ni), using a pcr-based approach employing degenerate primers designed on the basis of conserved regions of nuclear receptors, together with 5'- and 3'-race. an additional orphan nuclear receptor, hr4 fragment, was cloned from high five cells. comparison of these fragments with manduca sexta counterparts showed that the cloned sfecr [ecdysone ... | 2002 | 12020840 |
| insect cells and their potential as stabilization barriers for dna of multiple and single nucleopolyhedroviruses against ultraviolet-b-simulated sunlight inactivation. | a cell line from trichoplusia ni (tn-cl1) infected with the autographa californica multiple nucleopolyhedrovirus (acmnpv-hpp) and a cell line from helicoverpa zea (bcirl-hz-am1) infected with the helicoverpa zea single nucleopolyhedrovirus (hzsnpv/brcl2) were subjected to ultraviolet-b (uv-b) irradiation at a predetermined level of exposure that would inactivate greater than 95% of the virus suspended in the liquid. the working hypothesis was that the homologous insect cells would utilize their ... | 2002 | 12026166 |
| an azurocidin-like protein is induced in trichoplusia ni larval gut cells after bacterial challenge. | trichoplusia ni immune genes up-regulated in response to bacterial infection have been isolated using differential display polymerase chain reaction. here we report the cloning and characterisation of a gut-specific immune gene encoding an azurocidin-like protein. the deduced protein is 317 amino acid residues long with a hydrophobic c-terminus and a predicted 17-residue signal peptide. the mature t. ni protein shows 30% identity to human azurocidin, an antibacterial protein. like azurocidin, th ... | 2002 | 12031410 |
| expression and functional analysis of an inhibitor of apoptosis protein from trichoplusia ni. | an inhibitor of the apoptosis protein (iap) family gene from trichoplusia ni, tn-iap1v, a variant of lepidopteran tn-iap1, was cloned by rt-pcr. there are six single nucleotide polymorphisms between the two tn-iap1 variants, resulting in three predicted single amino acid polymorphisms. with the gst fusion expression system, soluble recombinant tn-iap1v was highly expressed in escherichia coli and then purified by affinity chromatography. caspase inhibition assays indicated that recombinant tn-ia ... | 2002 | 12054521 |
| storage stability of anagrapha falcifera nucleopolyhedrovirus in spray-dried formulations. | a multiply embedded nucleopolyhedrovirus isolated from anagrapha falcifera (kirby) (afmnpv) can lose insecticidal activity during months of dry storage in ambient room conditions. we tested the spray-dried afmnpv formulations after storage for up to 1 year at room temperatures for insecticidal activity against neonate trichoplusia ni (hübner). experimental formulations were made using combinations of corn flours, lignin, and sucrose, and were selected based on previous work which demonstrated th ... | 2002 | 12054782 |
| vertical transmission of tnsnpv, tncpv, acmnpv, and possibly recombinant npv in trichoplusia ni. | four viruses were tested for vertical transmission in trichoplusia ni: t. ni nucleopolyhedrovirus (tnsnpv), t. ni cypovirus (tncpv), autographa californica nucleopolyhedrovirus (acmnpv), and acmnpv engineered to express a scorpion toxin (acmnpv.aait). fifth instars were exposed to each virus, the survivors were reared and mated, and second-generation (f(1)) insects were examined for infection. tnsnpv was transmitted to offspring at a prevalence rate of 15.4%, tncpv at 10.2%, and acmnpv at 10.1%. ... | 2002 | 12054786 |
| 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 |
| signals involved in arabidopsis resistance to trichoplusia ni caterpillars induced by virulent and avirulent strains of the phytopathogen pseudomonas syringae. | plants have evolved different but interconnected strategies to defend themselves against herbivorous insects and microbial pathogens. we used an arabidopsis/pseudomonas syringae pathosystem to investigate the impact of pathogen-induced defense responses on cabbage looper (trichoplusia ni) larval feeding. arabidopsis mutants [npr1, pad4, eds5, and sid2(eds16)] or transgenic plants (nahg) that are more susceptible to microbial pathogens and are compromised in salicylic acid (sa)-dependent defense ... | 2002 | 12068100 |
| persistence of an occlusion-negative recombinant nucleopolyhedrovirus in trichoplusia ni indicates high multiplicity of cellular infection. | we use data from the serial passage of co-occluded recombinant autographa californica nuclear polyhedrosis virus (acmnpv) to estimate the viral multiplicity of infection of cells within infected insects. co-occlusion, the incorporation of wild-type and mutant virus genomes in the same occlusion body, has been proposed as a strategy to deliver genetically modified viruses as insecticides in a way that contains their spread in the environment. it may also serve as a means whereby naturally occurri ... | 2001 | 11679346 |
| lack of involvement of haemocytes in the establishment and spread of infection in spodoptera frugiperda larvae infected with the baculovirus autographa californica m nucleopolyhedrovirus by intrahaemocoelic injection. | it is thought that insect haemocytes, or blood cells, play an important role in baculovirus pathogenesis by amplifying and helping to spread the infection within the insect. here, infection is described of the lepidopteran noctuid species spodoptera frugiperda with the baculovirus autographa californica m nucleopolyhedrovirus (acmnpv). late instar s. frugiperda larvae were infected by intrahaemocoelic injection using a recombinant of acmnpv expressing the enhanced green fluorescent protein gene ... | 2002 | 12075074 |
| complex-type biantennary n-glycans of recombinant human transferrin from trichoplusia ni insect cells expressing mammalian [beta]-1,4-galactosyltransferase and [beta]-1,2-n-acetylglucosaminyltransferase ii. | a novel recombinant baculovirus expression vector was used to produce his-tagged human transferrin in a transformed insect cell line (tn5beta4galt) that constitutively expresses a mammalian beta-1,4-galactosyltransferase. this virus encoded the his-tagged human transferrin protein in conventional fashion under the control of the very late polyhedrin promoter. in addition, to enhance the synthesis of galactosylated biantennary n-glycans, this virus encoded human beta-1,2- n-acetylglucosaminyltran ... | 2003 | 12634321 |
| enzyme kinetics and glycan structural characterization of secreted alkaline phosphatase prepared using the baculovirus expression vector system. | secreted human alkaline phosphatase (seap, a model protein containing a single n-glycan chain) was expressed in spodoptera frugiperda sf-9 (sf-9) and trichoplusia ni bti-tn-5b1-4 (tn-5b1-4) insect cell lines infected with recombinant autographa californica multiple nuclear polyhedrovirus expressing seap under control of the polyhedrin promoter. sds-page showed that both systems expressed fairly pure rseap products. the rseap expression level was 7.0 u/ml in tn-5b1-4, higher than the 4.1 u/ml pro ... | 2002 | 12109816 |
| deletions in the ac-iap1 gene of the baculovirus acmnpv occur spontaneously during serial passage and confer a cell line-specific replication advantage. | the psti-i region of the autographa californica multicapsid nucleopolyhedrovirus (acmnpv) genome was previously shown to be a frequent target of spontaneous deletions during serial virus passage in tn-368 cells (kumar and miller, virus res. 7 (1987) 335). analysis of two of these serial passage mutants showed that a portion of the ac-iap1 gene was deleted. to directly test the effect of loss of ac-iap1, three different deletions in ac-iap1 were introduced into recombinant viruses and the ability ... | 2001 | 11682127 |
| evidence of oxidative stress following the viral infection of two lepidopteran insect cell lines. | the infection of spodoptera frugiperda sf-9 (sf-9) and trichoplusia ni bti-tn-5b1-4 (tn-5b1-4) insect cell lines with autographa californica multiple nucleopolyhedrovirus (acmnpv) resulted in increased levels of lipid hydroperoxides and protein carbonyls. in addition, the viral infection resulted in a significant decrease in the reduced glutathione to oxidized glutathione (2gsh/gssg) ratio. these results are all consistent with an increased level of oxidative stress as a result of the viral infe ... | 2001 | 11728817 |
| larval and pupal induction and n-terminal amino acid sequence of lysozyme from heliothis virescens. | fifth instar larvae and prepupae of heliothis virescens (tobacco budworm) were injected with live enterobacter cloacae and bled at different times after vaccination. immune pupal hemolymph showed a 54 times increase in lysozyme activity when compared with normal larval hemolymph, and an 11 times increase of lysozyme activity when compared with immune larval hemolymph. lysozyme activity of the normal pupal hemolymph increased as greatly as did lysozyme activity of the immune larval hemolymph afte ... | 2000 | 12770220 |
| detailed characterization of cysteine-less p-glycoprotein reveals subtle pharmacological differences in function from wild-type protein. | 1. subtle alterations in the coupling of drug binding to nucleotide hydrolysis were observed following mutation of all seven endogenous cysteine residues to serines in the human multidrug resistance transporter, p-glycoprotein. wild-type (wt) and the mutant (cys-less) forms of p-gp were expressed in trichoplusia ni (high five) cells and purified by metal affinity chromatography in order to undertake functional studies. 2. no significant differences were observed in substrate ([(3)h]-azidopine) b ... | 2001 | 11739236 |
| synthesis of new carboxylesterase inhibitors and evaluation of potency and water solubility. | carboxylesterases are essential enzymes in the hydrolysis and detoxification of numerous pharmaceuticals and pesticides. they are vital in mediating organophosphate toxicity and in activating many prodrugs such as the chemotherapeutic agent cpt-11. it is therefore important to study the catalytic mechanism responsible for carboxylesterase-induced hydrolysis, which can be accomplished through the use of potent and selective inhibitors. trifluoromethyl ketone (tfk)-containing compounds are the mos ... | 2001 | 11743738 |
| effect of silkworm hemolymph on n-linked glycosylation in two trichoplusia ni insect cell lines. | a recombinant n-linked glycoprotein, secreted human placental alkaline phosphatase (seap), was produced in two trichoplusia ni insect cell lines using the baculovirus expression vector. silkworm hemolymph (sh) was added to tnmfh + 10% fetal bovine serum (fbs) medium to a concentration of 2.5% or 5%, and seap production and glycosylation in the presence of sh were compared with controls devoid of hemolymph. growing tn-4s cells in 5% sh-supplemented medium required progressive adaptation of the ce ... | 2003 | 12889034 |
| acyl-coa z9- and z10-desaturase genes from a new zealand leafroller moth species, planotortrix octo. | two cdnas encoding acyl-coa z9-desaturase from the fat body and z10-desaturase from the pheromone gland of the greenhead leafroller moth, planotortrix octo, were obtained by race pcr. the z9-desaturase (pocto-z9) cdna spans 2291 nt with an orf encoding a 352 amino-acid protein, which has 65% identity to trichoplusia ni delta 9 desaturase (tni-z9). the z10-desaturase (pocto-z10) cdna spans 2777 nt with an orf encoding a protein with 356 amino acids. pocto-z10 shows lower identity to pocto-z9 and ... | 2002 | 12213232 |
| use of the piggybac transposon for germ-line transformation of insects. | germ-line transformation of insects is now possible with four independent transposable element vector systems. among these, the ttaa-insertion site specific transposon, piggybac, discovered in trichoplusia ni, is one of the most widely used. transformations have been achieved in a wide variety of dipterans, lepidopterans, and a coleopteran, and for many species, piggybac transposition was first tested by plasmid-based mobility assays in cell lines and embryos. all plasmid and genomic insertions ... | 2002 | 12225912 |
| insect control and dosage effects in transgenic canola containing a synthetic bacillus thuringiensis cryiac gene. | zygotic hypocotyls of canola (brassica napus l.) cv oscar, cv westar, and the breeding line uga188-20b were transformed with a truncated synthetic bacillus thuringiensis insecticidal crystal protein gene (bt cryiac) under the control of the cauliflower mosaic virus 35s promoter using agrobacterium tumefaciens-mediated transformation. fifty-seven independently transformed lines were produced, containing 1 to 12 copies of the transgenes. a range of cry expressors was produced from 0 to 0.4% cry as ... | 1996 | 12226379 |
| baculovirus lef-12 is not required for viral replication. | the baculovirus lef-12 (orf41) gene is required for transient expression of baculovirus late genes. to analyze the role of lef-12 in the context of infected cells, two mutant viruses were constructed. both mutants were viable in trichoplusia ni high 5 and spodoptera frugiperda sf9 cells. single-step growth curves, however, indicated that virus yields were reduced approximately fivefold in the absence of lef-12. pulse-labeling of infected cells revealed that lef-12 mutant viruses entered the late ... | 2002 | 12414945 |
| multitrophic interactions of the silverleaf whitefly, host plants, competing herbivores, and phytopathogens. | our laboratory found that silverleaf whitefly (slw; bemisia argentifolii bellows & perring) feeding alters host plant physiology and chemistry. the slw induces a number of host plant defenses, including pathogenesis-related (pr) protein accumulation (e.g., chitinases, beta-1,3-glucanases, peroxidases, chitosanases, etc.). induction of the pr proteins by slw feeding occurs in various plant species and varieties. the extent and type of induction is dependent on a number of factors that include hos ... | 2002 | 12432517 |
| the response of virally infected insect cells to dissolved oxygen concentration: recombinant protein production and oxidative damage. | the effects of dissolved oxygen (do) concentration on virally infected insect cells were investigated in 3-l bioreactor culture. specifically, cultures of spodoptera frugiperda sf-9 (sf-9) and trichoplusia ni bti-tn-5b1-4 (tn-5b1-4) were infected with autographa californica multiple nucleopolyhedrovirus expressing secreted alkaline phosphatase (seap). following infection at a do concentration of 50% air saturation, the do concentration was adjusted to a final value of either 190%, 50%, or 10% ai ... | 2003 | 12432586 |
| beta-(1 --> 4)-galactosyltransferase activity in native and engineered insect cells measured with time-resolved europium fluorescence. | to evaluate the ability of insect cells to produce complex-type n-glycans, beta-(1 --> 4)-galactosyltransferase (beta4galt) activity in several insect cell lines was analyzed. for this purpose, we developed a simple and highly sensitive assay for beta-(1 --> 4)-galactosyltransferase (beta4galt) activity, which is based on time-resolved fluorometry of europium. bovine serum albumin (bsa) modified with glcnac (glcnac(44)-bsa) was used as the acceptor. glcnac(44)-bsa was coated on a 96-well micropl ... | 2002 | 12433481 |
| recombinantly expressed isoenzymic aminopeptidases from helicoverpa armigera (american cotton bollworm) midgut display differential interaction with closely related bacillus thuringiensis insecticidal proteins. | several investigators have independently identified membrane-associated aminopeptidases in the midgut of insect larvae as the initial interacting ligand to the insecticidal crystal proteins of bacillus thuringiensis. though several isoenzymes of aminopeptidases have been identified from the midgut of an insect and their corresponding cdna cloned, only one of the isoform has been expressed heterologously and studied for its binding to cry toxins. here we report the cloning and expression of two a ... | 2003 | 12441000 |
| improved production of recombinant tumstatin in stably transformed trichoplusia ni bti tn 5b1-4 cells. | we describe the expression and in vitro activity of recombinant tumstatin from stably transformed trichoplusia ni bti tn 5b1-4 cells. recombinant tumstatin was secreted into a culture medium with a molecular weight of 29 kda. recombinant tumstatin was also purified to homogeneity using a simple one-step ni2+ affinity fractionation. purified recombinant tumstatin inhibited endothelial cell proliferation in a dose-dependent manner. the concentration at half-maximum inhibition (ed50) for recombinan ... | 2004 | 15039068 |
| natural occurrence of bacillus thuringiensis on cabbage foliage and in insects associated with cabbage crops. | bacillus thuringiensis was isolated from the phylloplane of organically grown cabbage in one field during two growth seasons (1992-93). the frequency of b. thuringiensis varied between 0.02 and 0.67 of the total b. cereus/b. thuringiensis population, with an average of 0.11. characterization of the b. thuringiensis isolates from foliage showed that the majority (64% of 150 isolates) belonged to serovar kurstaki, had bipyramidal crystals and toxicity towards pieris brassicae and/or trichoplusia n ... | 1997 | 12452602 |
| autographa californica m nucleopolyhedrovirus early gp64 synthesis mitigates developmental resistance in orally infected noctuid hosts. | the unusual early synthesis of the autographa californica m nucleopolyhedrovirus (acmnpv) budded virus (bv) structural protein gp64 is an important virulence factor during oral infection of heliothis virescens larvae. considering the breadth of the acmnpv host range, the importance of early gp64 synthesis in orally infected permissive hosts (trichoplusia ni and spodoptera exigua) from subfamilies other than that of h. virescens was assessed. an acmnpv recombinant, having wild-type early and late ... | 2004 | 15039526 |
| epitope mapping of human prostate specific antigen and glandular kallikrein expressed in insect cells. | two human prostate gland proteases were expressed in insect cells using recombinant baculovirus expression system. prostate specific antigen (psa) is an established serum marker of prostate cancer whereas the clinical utility of its close homologue, human glandular kallikrein (hk2) is presently unknown. the production levels using trichoplusia ni cells were roughly 300 &mgr;g/l and 6 mg/l for hk2 and psa, respectively. on western-blot we estimated the size for both proteins to be approximately 3 ... | 1997 | 12496928 |
| enterotoxigenicity and cytotoxicity of bacillus thuringiensis strains and development of a process for cry1ac production. | bacillus thuringiensis is indistinguishable from bacillus cereus except for the production of insecticidal crystal proteins (icps). b. thuringiensis strains may show enterotoxin profiles and toxin levels similar to those of b. cereus strains isolated from food-poisoning cases. it is important for the food industry and farmers to consider that with the application of b. thuringiensis strains to crops, their spores may be introduced into the human food chain. in this study, 59 b. thuringiensis str ... | 2003 | 12502392 |
| characterization of mamestra configurata nucleopolyhedrovirus enhancin and its functional analysis via expression in an autographa californica m nucleopolyhedrovirus recombinant. | enhancin genes have been identified in a number of baculoviruses and enhancin proteins are characterized by their ability to enhance the oral infectivity of heterologous baculoviruses in various lepidopteran insects. here, we describe the putative enhancin gene from mamestra configurata nucleopolyhedrovirus (maconpv), only the second npv in which an enhancin-like orf has been identified. the putative enhancin gene from maconpv has a typical baculovirus late promoter (ataag) 15 bp upstream from t ... | 2003 | 12533708 |
| comparison of the amino acid sequences of rna-dependent rna polymerases of cypoviruses in the family reoviridae. | the nucleotide sequences of the genome segment s2 of bombyx mori cypovirus 1, s2 of lymantria dispar cypovirus 1, s1 of lymantria dispar cypovirus 14 and s1 of a proposed new electropherotype of trichoplusia ni cypovirus 15 were determined. these segments encoded putative rna-dependent rna polymerases (rdrps). the deduced amino acid sequences of rdrps within the genus cypovirus showed 32% to 94% identities, while extent of homology between rdrps in the genera cypovirus and oryzavirus, a genus mo ... | 2003 | 12556988 |
| improving glucose and glutamine metabolism of human hek 293 and trichoplusia ni insect cells engineered to express a cytosolic pyruvate carboxylase enzyme. | metabolic engineering has been defined as a directed improvement of product formation or cellular properties by modification of specific biochemical pathways or introduction of new enzymatic reactions by recombinant dna technology. the use of metabolic flux analysis (mfa) has helped in the understanding of the key limitation in the metabolic pathways of cultured animal cells. the mfa of the major nutrients glucose and glutamine showed that the flux of glucose to the tca cycle and its subsequent ... | 2003 | 12573011 |
| production of a sialylated n-linked glycoprotein in insect cells: role of glycosidases and effect of harvest time on glycosylation. | using a nonengineered trichoplusia ni insect cell line, tn-4s, infected with an autographa californica recombinant baculovirus, 20% sialylation of human secreted placental alkaline phosphatase (seap) was observed. in contrast to this level of sialylation, intermediate complex forms with terminal galactose or n-acetylglucosamine were found in low proportions (<3% and <1%, respectively). we tested whether time of harvest or degradation of intermediate complex forms is responsible for this distribu ... | 2003 | 12573025 |
| improving baculovirus resistance to uv inactivation: increased virulence resulting from expression of a dna repair enzyme. | the use of baculoviruses as biological control agents is hampered by their susceptibility to inactivation by ultraviolet (uv) light. in an attempt to reduce uv inactivation, an algal virus pyrimidine dimer-specific glycosylase, cv-pdg, was expressed in the baculovirus autographa californica m nucleopolyhedrovirus (acmnpv), and the infectivity of recombinant viruses expressing cv-pdg was measured after exposure to uv light. expression of cv-pdg resulted in a 3-fold decrease in inactivation of bud ... | 2003 | 12581719 |
| dna microarrays of baculovirus genomes: differential expression of viral genes in two susceptible insect cell lines. | we describe, for the first time, the generation of a viral dna chip for simultaneous expression measurements of nearly all known open reading frames (orfs) in the best-studied members of the family baculoviridae, autographa californica multiple nucleopolyhedrovirus (acmnpv) and bombyx mori nucleopolyhedrovirus (bmnpv). in this study, a viral dna chip (ac-bmnpv chip) was fabricated and used to characterize the viral gene expression profile for acmnpv in different cell types. the viral chip is com ... | 2003 | 12607109 |
| production of recombinant human osteoprotegrin from trichoplusia ni cells and bombyx mori larvae. | human osteoprotegrin (opg) and its truncated mutant opg-280 and lengthened mutant opg-fc were constructed and successfully expressed in trichoplusia ni cells and bombyx mori larvae. native sds-page and western blot analysis revealed that opg-fc is present as a homodimer in tn cells or b. mori larvae compared with opg and opg-280. furthermore, the hypocalcemic effect assay showed that truncation of the c-terminal 100 residues opg does not abolish the biological activity and fc can be helpful in f ... | 2004 | 15327363 |
| in vitro expression and biochemical characterization of juvenile hormone esterase from manduca sexta. | juvenile hormone esterase (jhe) is a selective enzyme that hydrolyzes the methyl ester of juvenile hormone. this enzyme plays an important role in the regulation of metamorphosis in caterpillars, and is implicated in additional roles in development and reproduction in this and other orders of insect. the full length coding region of the jhe cdna from manduca sexta was subcloned into the baculovirus acmnpv genome under the control of the p10 promoter. the recombinant virus demonstrated the expres ... | 2003 | 12609517 |
| a few-polyhedra mutant and wild-type nucleopolyhedrovirus remain as a stable polymorphism during serial coinfection in trichoplusia ni. | few-polyhedra (fp) mutants of nucleopolyhedroviruses (npvs) are a well-known phenomenon during serial passage of virus in cell culture. under these circumstances such mutants produce low yields of occlusion bodies (obs) and poorly occlude virions, but they are selected for through advantageous rates of budded virus replication. spontaneous insertion of transposable elements originating from host cell dna into the viral fp25 gene has been shown to be a common cause of the phenotype. a model of np ... | 2003 | 12676682 |
| expression and assembly of arabidopsis thaliana pyruvate dehydrogenase in insect cell cytoplasm. | a vector was constructed for expression of arabidopsis thaliana mitochondrial pyruvate dehydrogenase (e1) in the cytoplasm of trichoplusia ni cells. the construct pddr101 comprises the mature-e1alpha coding sequence under control of the polh promoter, plus the mature-e1beta coding sequence under control of the p10 promoter. the e1alpha sequence was engineered to include an n-terminal his-tag. when protein samples were subjected to immobilized metal ion affinity chromatography, the alpha- and bet ... | 2003 | 12699701 |
| ex vivo monitoring of protein production in baculovirus-infected trichoplusia ni larvae with a gfp-specific optical probe. | trichoplusia ni larvae were infected with baculoviruses containing genes for the expression of ultraviolet optimized green fluorescent protein (gfpuv) and several product proteins. a gfp-specific optical probe was used to both excite the green fluorescent protein (lambda(ex) = 385 nm), and subsequently monitor fluorescence emission (lambda(em) = 514 nm) from outside the infected larvae. the probe's photodetector was connected to a voltmeter, which was used to quantify the amount of gfpuv express ... | 2003 | 12768630 |
| a baculovirus enhancin alters the permeability of a mucosal midgut peritrophic matrix from lepidopteran larvae. | the peritrophic matrix (pm) in lepidopterous larvae may function as a defensive barrier against ingested viral pathogens. pms isolated from trichoplusia ni and pseudaletia unipuncta larvae, were treated with a baculovirus-encoded metalloprotease (enhancin) from trichoplusia ni granulosis virus (tngv) and their in vitro permeability to blue dextran and fluorescent-labelled autographa californica nuclear polyhedrosis virus (acmnpv) was determined using a dual chamber permeability apparatus. incuba ... | 1999 | 12770384 |
| effect of culture conditions on the degree of sialylation of a recombinant glycoprotein expressed in insect cells. | secreted human placental alkaline phosphatase (seap) was produced in a nonengineered trichoplusia ni insect cell line, tn-4s, using a recombinant autographa californica baculovirus expression vector. the effect of culture conditions on seap specific yield and glycosylation was studied. when cultured in the high aspect ratio vessel (harv) or in tissue culture flasks (t-flasks), baculovirus-infected tn-4s cells produced high levels of seap (13 and 23 u/10(6) cells, respectively; 4 days postinfecti ... | 2003 | 12790633 |
| heterologous expression, purification, and enzymatic activity of mycobacterium tuberculosis nad(+) synthetase. | the enzyme nad(+) synthetase (nade) catalyzes the last step of nad biosynthesis. given nad vital role in cell metabolism, the enzyme represents a valid target for the development of new antimycobacterial agents. in the present study we expressed and purified two putative forms of mycobacterium tuberculosis nad(+) synthetase, differing in the polypeptide chain length (nade-738 and nade-679). furthermore, we evaluated several systems for the heterologous expression and large scale purification of ... | 2002 | 12182838 |
| rna interference as a metabolic engineering tool: potential for in vivo control of protein expression in an insect larval model. | many ex vivo factors influence the yield of recombinant protein produced via acmnpv (autographa californica multiple nucleocapsid nuclear polyhedrosis virus) in trichoplusia ni (t. ni) larvae. among these are: the method of infection, the time of infection, the virus load, and the time of harvest. in vivo strategies, however, that attempt to manipulate host function in this and other expression systems have largely been ignored. in this work, rna interference (rnai) is shown as an effective meta ... | 2003 | 12948752 |
| molecular evolutionary analysis of the widespread piggybac transposon family and related "domesticated" sequences. | piggybac is a short inverted-repeat-type dna transposable element originally isolated from the genome of the moth trichoplusia ni. it is currently the gene vector of choice for the transformation of various insect species. a few sequences with similarity to piggybac have previously been identified from organisms such as humans ( looper), the pufferfish takifugu rubripes ( pigibaku), xenopus ( tx), daphnia ( pokey), and the oriental fruit fly bactrocera dorsalis. we have now identified 50 piggyba ... | 2003 | 12955498 |
| pseudomonas syringae manipulates systemic plant defenses against pathogens and herbivores. | many pathogens are virulent because they specifically interfere with host defense responses and therefore can proliferate. here, we report that virulent strains of the bacterial phytopathogen pseudomonas syringae induce systemic susceptibility to secondary p. syringae infection in the host plant arabidopsis thaliana. this systemic induced susceptibility (sis) is in direct contrast to the well studied avirulence/r gene-dependent resistance response known as the hypersensitive response that elicit ... | 2005 | 15657122 |
| field activity and storage stability of anagrapha falcifera nucleopolyhedrovirus (afmnpv) in spray-dried lignin-based formulations. | a multiple-embedded nucleopolyhedrovirus isolated from anagrapha falcifera (kirby) (afmnpv) has potential to be developed into a microbial bioinsecticide because the host range includes several economic pests. we tested spray-dried afmnpv formulations after storage for insecticidal activity based on bioassays with neonate trichoplusia ni (hübner). eight experimental lignin-based spray-dried formulations, a glycerin-based formulation, and an unformulated sample were made with virus stock from thr ... | 2003 | 14503576 |
| rna interference silences microplitis demolitor bracovirus genes and implicates glc1.8 in disruption of adhesion in infected host cells. | the family polydnaviridae consists of ds-dna viruses that are symbiotically associated with certain parasitoid wasps. pdvs are transmitted vertically but also are injected by wasps into hosts where they cause several physiological alterations including immunosuppression. the pdv genes responsible for mediating immunosuppression and other host alterations remain poorly characterized in large measure because viral mutants cannot be produced to study gene function. here we report the use of rna int ... | 2003 | 14554081 |
| rapid evolution and the cost of resistance to bacillus thuringiensis in greenhouse populations of cabbage loopers, trichoplusia ni. | the microbial insecticide bacillus thuringiensis (bt) has become the mainstay of non-chemical control of lepidopteran pests, either as sprays or through the incorporation of bt toxins into transgenic crops. given the wide use of bt, it is striking that currently only one pest species, plutella xylostella, has been reported to have developed significant resistance to bt outside the laboratory. by contrast, we report here the frequent and rapid development of resistance to b. thuringiensis kurstak ... | 2003 | 14613613 |
| synergism between the nucleopolyhedroviruses of autographa californica and trichoplusia ni. | previous observations on high virulence of autographa californica multiple nucleopolyhedrovirus (acmnpv) and trichoplusia ni single nucleopolyhedrovirus (tnsnpv) acting together led us to test possible synergism between these two nucleopolyhedroviruses (npvs) on cabbage looper larvae. because synergism between acmnpv and the trichoplusia ni granulovirus (tngv) has been well established before, these two viruses were included in this study as a positive control. each virus was assayed separately ... | 2003 | 14658849 |
| heterologous expression of maize (zea mays l.) mir1 cysteine proteinase in eukaryotic and prokaryotic expression systems. | several heterologous expression systems were tested for their ability to express a unique maize cysteine proteinase mir1. a baculovirus-based expression system using trichoplusia ni larvae as host resulted in the expression of mir1 that was correctly processed and exhibited proteinase activity. expression in escherichia coli resulted in accumulation of mir1, but it had limited solubility and enzymatic activity. large quantities of mir1 were produced when pichia pastoris was used as the host, but ... | 2004 | 14766309 |
| deletion of the autographa californica nucleopolyhedrovirus chitinase kdel motif and in vitro and in vivo analysis of the modified virus. | infection of insect larvae with autographa californica nucleopolyhedrovirus (acmnpv) results in the liquefaction of the host, a process involving the action of virus-encoded chitinase and cathepsin gene products. chitinase is localized in the endoplasmic reticulum (er) during infection because of the presence of a c-terminal er retrieval motif (kdel). in this study, the kdel coding region was removed from the chitinase gene so that expression of the modified chitinase remained under the control ... | 2004 | 15039525 |
| metabolism of thymol and trans-anethole in larvae of spodoptera litura and trichoplusia ni (lepidoptera: noctuidae). | metabolism of the monoterpenoid thymol and the phenylpropanoid trans-anethole, both constituents of essential oils, was investigated following topical and oral administration of the compounds to the tobacco cutworm (spodoptera litura) and the cabbage looper (trichoplusia ni). in both species and irrespective of route, administration of thymol resulted in the excretion of its 3-o-beta-glucoside, whereas trans-anethole was hydroxylated on the side chain methyl group. both metabolites were isolated ... | 2004 | 15113155 |
| crystallization of glycosylated human bace protease domain expressed in trichoplusia ni. | human beta-amyloid precursor protein cleaving enzyme (beta-secretase, or bace) belongs to the aspartyl protease family, and is responsible for generating the n-terminus of beta-amyloid peptide (abeta). bace is a type i transmembrane glycoprotein with pre-, pro- and catalytic domains, a short transmembrane helix and a cytoplasmic region. in this study, a truncated form was engineered to produce the authentic catalytic domain of bace in trichoplusia ni (high 5) cells. the glycosylated bace zymogen ... | 2004 | 15134659 |
| effects of ac150 on virulence and pathogenesis of autographa californica multiple nucleopolyhedrovirus in noctuid hosts. | ac150 is expressed late during infection of cultured lepidopteran insect cells by autographa californica multiple nucleopolyhedrovirus. the ac150 gene product is predicted to have a molecular mass of 11 161 da and consists of a hydrophobic n terminus and a single 'peritrophin-a'-like domain, connected by a short region of charged amino acids. an ac150 deletion mutant and its parental wild-type virus were compared for differences in virulence by both oral and intrahaemocoelic routes of infection. ... | 2005 | 15914839 |
| characterization of two autographa californica nucleopolyhedrovirus proteins, ac145 and ac150, which affect oral infectivity in a host-dependent manner. | the genome of the baculovirus autographa californica nuclear polyhedrosis virus (acmnpv) contains two homologues, orf145 and orf150, of the heliothis armigera entomopoxvirus (haepv) 11,000-kda gene. polyclonal antibodies raised against the ac145 or ac150 protein were utilized to demonstrate that they are expressed from late to very late times of infection and are within the nuclei of infected sf-21 cells. transmission electron microscopy coupled with immunogold labeling of ac145 found this prote ... | 2004 | 15163737 |
| p74 mediates specific binding of autographa californica m nucleopolyhedrovirus occlusion-derived virus to primary cellular targets in the midgut epithelia of heliothis virescens larvae. | p74, an envelope protein of the occlusion-derived virus (odv) of autographa californica m nucleopolyhedrovirus (acmnpv), is critical for oral infection of trichoplusia ni larvae. the role of p74 during primary infection, however, is unknown. here we provide evidence that p74 facilitates binding of acmnpv odv to a specific receptor within the larval midgut epithelia of another host species, heliothis virescens. we adapted a fluorescence dequenching assay to compare binding, fusion, and competitio ... | 2004 | 15194753 |
| identification and characterization of a major early-transcribed gene of trichoplusia ni single nucleocapsid nucleopolyhedrovirus using the baculovirus expression system. | an early transcribed gene (me-53) of a south africa strain of trichoplusia ni single nucleocapsid nucleopolyhedrovirus (tnsnpv) was sequenced and identified. it has an open reading frame of 1146 nucleotides that encodes a protein of 382 amino acids with a molecular mass of 45.2 kda. the deduced protein sequence alignment of 13 baculovirus me-53s indicated that the tnsnpv me-53 shares the highest homologies with npv subgroup ii-a spodoptera exigua multiple and mamestra configurata (maco) nucleopo ... | 2004 | 15215681 |
| molecular characterization of the major virion protein gene from the trichoplusia ni ascovirus. | ascoviruses (avs) belong to a family of double-stranded dna viruses that infect lepidoptera insects and cause the unique pathology of forming virion-containing vesicles in the hemolymph of infected hosts. virions of avs are large and contain more than 12 polypeptides. a gene, tnav-cp, encoding the major structural protein of the trichoplusia ni ascovirus 2a (tnav-2a) was cloned by immunoscreening an expression library with antibodies against total tnav virion proteins. tnav-cp is an intronless g ... | 2003 | 12913362 |
| life history and cost analysis for continuous rearing of perillus bioculatus (heteroptera: pentatomidae) on a zoophytogenous artificial diet. | the impact of a zoophytogenous, insect-free artificial diet on the developmental rate, life history parameters, and fertility was examined over 11 consecutive generations for domesticated perillus biocullatus (f.) (heteroptera: pentatomidae). this study showed that when fed an insect-free artificial diet during both the nymphal and adult stages, developmental times were prolonged, and the net reproductive rates (r0) and the intrinsic rates of increase (r(m)) were significantly lower than when fe ... | 2004 | 15279257 |
| differentially enhanced insect resistance, at a cost, in arabidopsis thaliana constitutively expressing a transcription factor of defensive metabolites. | a transgenic line of arabidopsis thaliana constitutively expressing a conserved myb transcription factor of phenylpropanoid biosynthesis resulting in solid-purple leaves had significantly increased resistance to leaf feeding by first instar fall armyworms (spodoptera frugiperda), but no enhanced resistance to cabbage looper (trichoplusia ni) larvae, when compared to wild type plants. however, inflorescence and silique (seed pod) production were significantly reduced by 22 and 52%, respectively, ... | 2004 | 15291486 |
| [characterization of resistant cell line of trichoplusia ni to bacillus thuringiensis toxin cry1ac]. | the cell line (bti-tn-5b1-4) of trichoplusia ni had been selected by bacilllus thuringiensis (bt) toxin cry1ac for 86 generations continuously. a high level of resistance was achieved and the partial characterizations of the resistance-selected bti-tn-5b1-4 cells were studied in order to explore the mechanism of the resistance to bt toxin. the cells selected by cry1ac had low level of cross-resistance (19.7 fold) to cry1c, their ability of tolerance in the hypotonic solution increased evidently ... | 2004 | 15323422 |
| improved pc1/3 production through recombinant expression in insect cells and larvae. | protein convertase 1/3 is a serine endoproteinase present in the regulated secretory pathway of endocrine and neuroendocrine cells. it is responsible for the processing of numerous prohormones and proneuropeptides into their biologically active moieties, often following cleavage at pairs of basic residues. the determination of its three-dimensional structure, as well as the understanding of its enzymatic properties, would greatly benefit from the production and availability of large amounts of r ... | 2004 | 15358360 |
| inheritance of resistance to bacillus thuringiensis subsp. kurstaki in trichoplusia ni. | the genetic inheritance of resistance to a commercial formulation of bacillus thuringiensis subsp. kurstaki was examined in a trichoplusia ni colony initiated from a resistant population present in a commercial vegetable greenhouse in british columbia, canada. progeny of f(1) reciprocal crosses and backcrosses between f(1) larvae and resistant (p(r)) and susceptible (p(s)) populations were assayed at different b. thuringiensis subsp. kurstaki concentrations. the responses of progeny of reciproca ... | 2004 | 15466525 |
| identification, characterization and phylogenic analysis of conserved genes within the odvp-6e/odv-e56 gene region of choristoneura fumiferana granulovirus. | the genes that are located within the odvp-6e/odv-e56 region of the choristoneura fumiferana granulovirus (chfugv) were identified by sequencing the 11 kb bamhi restriction fragment on the chfugv genome. the global gc content that was calculated from the data obtained from this genomic region was 34.96%. the open-reading frames (orfs), located within the odvp-6e/odv-e56 region, are presented and compared to the equivalent orfs that are located at the same region in other gvs. this region is comp ... | 2004 | 15469697 |
| expression, purification and sublocalization of sars-cov nucleocapsid protein in insect cells. | the causative agent of severe acute respiratory syndrome (sars) is a previously unidentified coronavirus, sars-cov. the nucleocapsid (n) protein of sars-cov is a major viral protein recognized by acute and early convalescent sera from sars patients. to facilitate the studies on the function and structure of the n protein, this report describe the expression and purification of recombinant sars-cov n protein using the baculovirus expression system. recombinant hexa-histidine-tagged n protein with ... | 2004 | 15514849 |
| effect of expression of manganese superoxide dismutase in baculovirus-infected insect cells. | it has previously been demonstrated that baculovirus infection of the spodoptera frugiperda sf-9 (sf-9) and trichoplusia ni bti-tn-5b1-4 (tn-5b1-4) insect cell lines leads to oxidative stress as measured by protein and membrane lipid oxidation and that this oxidative damage contributes to cell death. as a result of these findings, it was hypothesized that baculovirus infection stimulates superoxide radical (o(2)(.-)) synthesis in the mitochondria and that the resulting o(2)(.-) accumulation over ... | 2004 | 15531788 |
| mannosamine supplementation extends the n-acetylglucosaminylation of recombinant human secreted alkaline phosphatase produced in trichoplusia ni (cabbage looper) insect cell cultures. | a major limitation of the insect cell-baculovirus expression vector system is its poor capacity to perform complex glycosylation, since the glycoproteins produced are usually only of the high-mannose and paucimannose types. nonetheless, recent evidence indicates that, under various conditions, some insect cell lines are capable of producing complex-type glycans. in the present study, we assessed the effects of supplementation with mann (mannosamine) mannac (n-acetylmannosamine) and cyt (cytidine ... | 2005 | 15574126 |
| molecular cloning and functional characterization of a neuronal choline transporter from trichoplusia ni. | a cdna encoding a high-affinity na(+)-dependent choline transporter (trncht) was isolated from the cns of the cabbage looper trichoplusia ni using an rt-pcr-based approach. the deduced amino acid sequence of the cht cdna predicts a 594 amino acid protein of 64.74 kda prior to glycosylation. trncht has 80%, 79%, 76%, and 58% amino acid identity to putative chts from anopheles gambiae, drosophila melanogaster and apis mellifera, and a cloned cht from limulus polyphemus, respectively. in situ hybri ... | 2005 | 15607656 |
| immunofluorescence assay for detection of human metapneumovirus-specific antibodies by use of baculovirus-expressed fusion protein. | human metapneumovirus (hmpv) has recently been identified as an etiological agent of acute respiratory infections. the hmpv fusion (f) protein has been indicated to be a major antigenic determinant that mediates effective neutralization and protection against hmpv infection. we developed a new immunofluorescence assay (ifa) using trichoplusia ni (tn5) insect cells infected with a recombinant baculovirus-expressing hmpv f protein (bac-f ifa). a total of 200 serum samples from japanese people 1 mo ... | 2005 | 15643008 |
| glc1.8 from microplitis demolitor bracovirus induces a loss of adhesion and phagocytosis in insect high five and s2 cells. | polydnaviridae is a unique family of dna viruses that are symbiotically associated with parasitoid wasps. upon oviposition, wasps inject these viruses into their hosts, where they cause several physiological alterations, including suppression of the cellular immune response. here we report that expression of the glc1.8 gene from microplitis demolitor bracovirus (mdbv) causes a loss of adhesion by two hemocyte-like cell lines, namely, high five cells from the lepidopteran trichoplusia ni and s2 c ... | 2005 | 15650210 |