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inheritance of resistance to bacillus thuringiensis cry1ac toxin in a greenhouse-derived strain of cabbage looper (lepidoptera: noctuidae).a population of cabbage looper, trichoplusia ni (hübner), collected from commercial greenhouses in the lower mainland of british columbia, canada, in 2001 showed a resistance level of 24-fold to dipel, a product of bacillus thuringiensis (bt) subspecies kurstaki. this population was selected with cry1ac, the major bt cry toxin in dipel, to obtain a homogenous population resistant to cry1ac. the resulting strain of t. ni, named glen-cry1ac, was highly resistant to cry1ac with a resistance ratio o ...200415666767
cloning and characterization of the g protein betagamma subunits from trichoplusia ni (high five cells).baculoviral-mediated expression in insect cells has become a method of choice where high-level protein expression is desired and where expression in escherichia coliform (e. coli.) is unsuitable. genes of interest are inserted into the baculoviral genome of autographa californica nuclear polyhedrosis virus (acnpv) under the extremely strong, but very late polyhedron gene (polh). the preferred host lines are derived from spodoptera frugiperda (sf9 or sf21) or tricoplusia ni (high five, invitrogen ...200515763469
biological and molecular characterization of a multicapsid nucleopolyhedrovirus from thysanoplusia orichalcea (l.) (lepidoptera: noctuidae).a multicapsid nucleopolyhedrovirus (thormnpv) that was co-isolated with a single nucleocapid thorsnpv from mixed infected larvae of thysanoplusia orichalcea l. (lepidoptea: noctuidae) is characterized. scanning electron microscopy of thormnpv showed a dodecahedral-shaped occlusion body (ob). the occluded virions contained one to as many as eight nucleocapsids/virion. virion band profiles in gradient centrifugation were consistent in at least 10 rounds of centrifugation from different virion samp ...200515766929
characterization of cultured insect cells selected by bacillus thuringiensis crystal toxin.selection for resistance against bacillus thuringiensis (bt) cry1ac10 in the trichoplusia ni (hubner) cell line bti-tn-5b1-4 (tnh5) was tested, and the development of resistance in the selected cells was like a s-form curve. monitoring at the cry1ac10 50th challenge, the resistance ratio was 1,294-fold as many as that of initial cells. but the resistance to cry1ac10 declined gradually when the selection was relaxed. the resistance declined rapidly at the low level of resistance and slowly at the ...200415780008
antimicrobial activity of conditioned medium fractions from spodoptera frugiperda sf9 and trichoplusia ni hi5 insect cells.concentrated conditioned medium (cm) fractions from spodoptera frugiperda sf9 and trichoplusia ni cells, eluting from a gel filtration column at around 10 kda, were found to exhibit strong antibacterial activity against bacillus megaterium and escherichia coli. the b. megaterium cells incubated in the cm fraction from sf9 cells rapidly lost viability: after 8 min the viability had decreased to 0.7%, as compared with the control. addition of the cm fraction to e. coli cells resulted in a less dra ...200515806353
insights into the central metabolism of spodoptera frugiperda (sf-9) and trichoplusia ni bti-tn-5b1-4 (tn-5) insect cells by radiolabeling studies.the insect cell baculovirus expression vector system (bevs) is one of the most commonly used expression systems for recombinant protein production. this system is also widely used for the production of recombinant virus and virus-like particles. although several published reports exist on recombinant protein expression using insect cells, information dealing with their metabolism in vitro is relatively scarce. in this work we have analyzed the metabolism of glucose and glutamine, the main carbon ...200515903243
production of recombinant adeno-associated viral vectors using a baculovirus/insect cell suspension culture system: from shake flasks to a 20-l bioreactor.production of recombinant adeno-associated viral vectors using a baculovirus/insect cell system at various scales is presented. shake flask studies were conducted to assess conditions to be used in bioreactors. two insect cell lines, trichoplusia ni (h5) and spodoptera frugiperda (sf9), were compared for their ability to produce raav-2 after infection with recombinant baculoviruses coding for the essential components of the vector. the effect of varying the ratio between individual baculoviruses ...200515903253
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. ...200515914839
baculovirus-based expression of an insect viral protein in 12 different insect cell lines.the ability of 12 unique lepidopteran insect cell lines from anticarsia gemmatalis, heliothis virescens, lymantria dispar (two lines), mamestra brassica, plutella xylostella, spodoptera frugiperda (two lines), and trichoplusia ni (three lines) to support production of a recombinant polydnavirus (pdv) protein (gipdv 1.1) expressed using the bac-to-bac baculovirus expression system was examined. polydnavirus gene gipdv 1.1 was cloned into the pfastbac baculovirus vector under the control of the po ...200515926859
arabidopsis iqd1, a novel calmodulin-binding nuclear protein, stimulates glucosinolate accumulation and plant defense.glucosinolates are a class of secondary metabolites with important roles in plant defense and human nutrition. to uncover regulatory mechanisms of glucosinolate production, we screened arabidopsis thaliana t-dna activation-tagged lines and identified a high-glucosinolate mutant caused by overexpression of iqd1 (at3g09710). a series of gain- and loss-of-function iqd1 alleles in different accessions correlates with increased and decreased glucosinolate levels, respectively. iqd1 encodes a novel pr ...200515960618
characterization of symmetric complexes of nerve growth factor and the ectodomain of the pan-neurotrophin receptor, p75ntr.nerve growth factor (ngf) is the ligand for two unrelated cellular receptors, trka and p75(ntr), and acts as a mediator in the development and maintenance of the mammalian nervous system. signaling through trka kinase domains promotes neuronal survival, whereas activation of the p75(ntr) "death domains" induces apoptosis under correct physiological conditions. however, co-expression of these receptors leads to enhanced neuronal survival upon ngf stimulation, possibly through a ternary p75(ntr) x ...200516009712
the cost of resistance to bacillus thuringiensis varies with the host plant of trichoplusia ni.selection for resistance to insecticides, diseases and parasitoids is assumed to be costly and often requires tradeoffs with reproductive fitness. the costs of resistance, however, are often difficult to measure. cabbage looper, trichoplusia ni, a generalist lepidopteran herbivore, has become highly resistant following the extensive use of the microbial insecticide, bacillus thuringiensis kurstaki (bt) in vegetable greenhouses. we compared the growth rate, pupal size and survival of resistant, s ...200516024361
comparative venom toxicity between pteromalus puparum and nasonia vitripennis (hymenoptera: pteromalidae) toward the hemocytes of their natural hosts, non-target insects and cultured insect cells.crude venoms from two parasitoid species, pteromalus puparum and nasonia vitripennis (hymenoptera: pteromalidae) were assayed for biological activities toward hemocytes from two species of their natural hosts and eight species of their non-natural hosts as well as two lines of cultured lepidoptera cells, respectively. by inhibiting the spreading and viability of insect hemocytes, the venom from p. puparum displayed significantly higher activities toward plasmatocytes and granular cells from both ...200516026808
ectopic expression of the human adenine nucleotide translocase, isoform 3 (ant-3). characterization of ligand binding properties.the adenine nucleotide translocase (ant) is a key component in maintaining cellular energy homeostasis, and has also been implicated in formation of the mitochondrial permeability transition pore. human ant-3 was cloned from a human heart cdna library and expressed as a histidine-tagged fusion protein in the mitochondria of the trichoplusia ni. cell line. overexpression resulted in a concomitant decrease in the endogenous ant content, allowing for the characterization of binding of known ant lig ...200516060289
efficient transposition of the piggybac (pb) transposon in mammalian cells and mice.transposable elements have been routinely used for genetic manipulation in lower organisms, including generating transgenic animals and insertional mutagenesis. in contrast, the usage of transposons in mice and other vertebrate systems is still limited due to the lack of an efficient transposition system. we have tested the ability of piggybac (pb), a dna transposon from the cabbage looper moth trichoplusia ni, to transpose in mammalian systems. we show that pb elements carrying multiple genes c ...200516096065
functional characterization of medaka cyp3a38 and cyp3a40: kinetics and catalysis by expression in a recombinant baculovirus system.phylogenic analysis of the teleost genomic lineages has demonstrated the precedent for multiple genome duplications. among many of the genes duplicated, cytochrome p450 genes have undergone independent diversification, which can be traced to a single ancestral gene. in teleosts, cytochrome p450s, from all major families, have been identified. among these, the cyp3a family has been cloned in several teleost species and demonstrated to contain multiple paralogs differing in gene expression pattern ...200516112913
assembly of homotrimeric type xxi minicollagen by coexpression of prolyl 4-hydroxylase in stably transfected drosophila melanogaster s2 cells.we established stably transfected insect cell lines containing cdnas encoding the alpha and beta subunits of human prolyl 4-hydroxylase in both trichoplusia ni and drosophila melanogaster s2 cells. the expression level and enzymatic activity of recombinant prolyl 4-hydroxylase produced in the drosophila expression system were significantly higher than those produced in the t. ni system. we further characterized the involvement of prolyl 4-hydroxylase in the assembly of the three alpha chains to ...200516115607
a novel chitin-binding protein identified from the peritrophic membrane of the cabbage looper, trichoplusia ni.a novel midgut peritrophic membrane (pm) protein, tnpm-p42, was identified from the cabbage looper, trichoplusia ni. tnpm-p42 was shown as a 42kda protein by sds-page analysis and appeared to be associated with the pm throughout its entire length. in t. ni larvae, the midgut is the only tissue where tnpm-p42 could be detected during the feeding period of the larvae. tnpm-p42 has chitin-binding activity and is strongly associated with the pm, which is similar to the currently known peritrophin ty ...200516203204
expression and characterization of recombinant beta-secretase from trichoplusia ni bti tn5b1-4 cells transformed with cdnas encoding human beta1,4-galactosyltransferase and gal beta1,4-glcnac alpha 2,6-sialytransferase.beta-secretase (betasec) was expressed in trichoplusia ni bti tn5b1-4 (tn5b1-4) cells transformed with cdnas encoding beta1,4-galactosyltransferase (galt) and gal beta1,4-glcnac alpha 2,6-sialyltransferase (st). the apparent molecular weight of recombinant beta-secretase was increased from 57 to 59 k da. a lectin blot analysis indicated that recombinant beta-secretase from tn5b1-4 betasec/galt-st cells (tn5b1-4 cells co-transformed with cdnas encoding beta-secretase, glycosyltransferases, galt, ...200516226891
transformed lepidopteran cells expressing a protein of the silkmoth fat body display enhanced susceptibility to baculovirus infection and produce high titers of budded virus in serum-free media.baculovirus vectors constitute important tools for therapeutic protein production and mammalian cell transduction for gene therapy applications. a prerequisite for such applications is that the cell lines in which baculoviruses are propagated be maintained in serum-free media that are devoid of potential human pathogens. however, in serum-free media, the performance of baculovirus-based systems can be significantly reduced. in this report, we show that silkmoth-derived host cell lines for the bo ...200516233927
yeast extract from express five serum-free medium contains factors at about 35 kda, essential for growth of trichoplusia ni insect cells.the yeast extract (of unknown origin) present in the commercially available serum-free medium 'express five' contains factors ('yeast extract factors') up to 35 kda which are essential for growth of trichoplusia ni insect cells. a yeast extract brand lacking these components could not support growth of t. ni cells. however, cell proliferation was restored by adding chromatographic fractions containing the yeast extract factors. the yeast extract factors were not solely responsible for the growth ...200516245184
factors influencing in vitro infectivity and growth of rickettsia peacockii (rickettsiales: rickettsiaceae), an endosymbiont of the rocky mountain wood tick, dermacentor andersoni (acari, ixodidae).rickettsia peacockii, a spotted fever group rickettsia, is a transovarially transmitted endosymbiont of rocky mountain wood ticks, dermacentor andersoni. this rickettsia, formerly known as the east side agent and restricted to female ticks, was detected in a chronically infected embryonic cell line, dae100, from d. andersoni. we examined infectivity, ability to induce cytopathic effect (cpe) and host cell specificity of r. peacockii using cultured arthropod and mammalian cells. aposymbiotic dae1 ...200516288906
high level expression of monomeric and dimeric human alpha1,3-fucosyltransferase v.alpha3/4-fucosyltransferases play a crucial role in inflammatory processes and tumor metastasis. while several human fucosyltransferases (fucts) with different acceptor substrate specificities have been identified, the design of specific inhibitors for therapeutic approaches is hampered by the lack of structural information. in this study, we evaluated the expression of different constructs of human fucosyltransferase v to generate the large amounts required for structural studies. the truncated ...200616290306
comparison of solid-phase radioimmunoassays for baculoviruses.the sensitivity and cross-reaction of four solid-phase radioimmunoassays (ria) for trichoplusia ni nuclear polyhedrosis virus containing singly enveloped virions were investigated. the detection limits of each assay were as follows: indirect ria, 5 ng of dissolved polyhedron antigen; direct ria, 50 ng; indirect sandwich ria, 200 ng; and direct sandwich ria, 300 ng. the indirect and indirect sandwich rias showed considerable cross-reaction with other baculovirus antigens, but the direct and direc ...197816345305
plaque assay of nuclear polyhedrosis viruses in cell culture.the nuclear polyhedrosis virus of autographa californica has been titrated in spodoptera frugiperda cells by the plaque method, using a solid overlay which does not require either the use of modified culture medium or expensive purified agarose or the addition of culture medium as a liquid layer above the solid agarose. this assay is more sensitive than that using a viscous methyl cellulose overlay but less sensitive than the end-point dilution technique. neither trichoplusia ni nor bombyx mori ...197816345306
hydrophobic interactions involved in attachment of a baculovirus to hydrophobic surfaces.the hydrophobic interactions of trichoplusia ni nuclear polyhedrosis virus were characterized by hydrophobic interaction chromatography. the determination of the hydrophobic force and some of the factors that influence its size is discussed in relation to the attachment to leaf surfaces of polyhedra during their use as biological control agents against insect pests.198616347113
cricket paralysis virus, a potential control agent for the olive fruit fly, dacus oleae gmel.representatives of several families of insect viruses were tested for growth and pathogenicity in the olive fruit fly, dacus oleae gmel. the viruses included nuclear polyhedrosis viruses, an iridovirus, two picornaviruses, and trichoplusia ni small rna virus (a member of the nudaurelia beta family), in addition to two naturally occurring viruses of the olive fruit fly. two viruses, one of the two picornaviruses (cricket paralysis virus [crpv] and the iridovirus (type 21 from heliothis armigera), ...198716347255
toxic trypsin digest fragment from the bacillus thuringiensis parasporal protein.enzymatic digestion in vitro of the bacillus thuringiensis protoxin presumably releases and activates the toxin in a manner analogous to that which occurs when a b. thuringiensis sporulated fermentation preparation passes through the midgut of a lepidopteran larva. therefore, a sporulated culture of b. thuringiensis subsp. kurstaki (serotype 3a3b) hd-263 was treated with trypsin to release an activated toxin soluble in bicarbonate buffer. a 63-kilodalton protein, toxic to cabbage looper larvae ( ...198716347290
measurement of surface charge of baculovirus polyhedra.the isoelectric points of three baculoviruses, trichoplusia ni nuclear polyhedrosis virus (npv), t. ni granulosis virus, and spodoptera littoralis npv were identified by cell electrophoresis. at neutral ph polyhedra were negatively charged. t. ni npv polyhedra were reacted with a number of reagents which could potentially attach to or degrade their surface structure. this gave information on the components that contribute to the charge profile of t. ni npv. this is discussed in relation to the u ...198716347308
co-occlusion and persistence of a baculovirus mutant lacking the polyhedrin gene.a co-occlusion process was evaluated as a commercially and ecologically acceptable strategy for the development of genetically improved baculovirus insecticides. coinfection of spodoptera frugiperda (iplb-sf-21) tissue culture cells with autographa californica nuclear polyhedrosis virus (acmnpv) and an acmnpv mutant (ac-e10) lacking the polyhedrin gene resulted in occlusion of both virus types within polyhedra. the amount of occluded ac-e10 virions in progeny polyhedra populations during serial ...199016348313
the phylloplane as a source of bacillus thuringiensis variants.novel variants of bacillus thuringiensis were isolated from the phylloplane of deciduous and conifer trees as well as of other plants. these isolates displayed a range of toxicity towards trichoplusia ni. immunoblot and toxin protein analysis indicate that these strains included representatives of the three principal b. thuringiensis pathotypes active against larvae of the orders lepidoptera, diptera, and coleoptera. we propose that b. thuringiensis be considered part of the common leaf microflo ...199116348400
binding of insecticidal crystal proteins of bacillus thuringiensis to the midgut brush border of the cabbage looper, trichoplusia ni (hübner) (lepidoptera: noctuidae), and selection for resistance to one of the crystal proteins.the susceptibility of trichoplusia ni larvae to several bacillus thuringiensis insecticidal crystal proteins (icps) was tested. neonatal larvae proved to be susceptible to solubilized trypsin-treated cryia(a), cryia(b), and cryia(c) (50% lethal concentrations [lc(50)s], 570, 480, and 320 ng/cm, respectively) but showed little susceptibility to cryib and cryid (lc(50)s, 5,640 and 2,530 ng/cm, respectively). the toxicity of icps was correlated to binding to the epithelial brush border of the midgu ...199416349420
molecular and biological characterization of a cypovirus from the mosquito culex restuans.a cypovirus from the mosquito culex restuans (named crcpv) was isolated and its biology, morphology, and molecular characteristics were investigated. crcpv is characterized by small (0.1-1.0 microm), irregularly shaped inclusion bodies that are multiply embedded. laboratory studies demonstrated that divalent cations influenced transmission of crcpv to culex quinquefasciatus larvae; magnesium enhanced crcpv transmission by approximately 30% while calcium inhibited transmission. crcpv is the secon ...200616376932
dual mutations in the autographa californica nucleopolyhedrovirus fp-25 and p35 genes result in plasma-membrane blebbing in trichoplusia ni cells.spodoptera frugiperda cells infected with autographa californica nucleopolyhedrovirus (acmnpv) lacking a functional anti-apoptotic p35 protein undergo apoptosis. however, such mutants replicate normally in trichoplusia ni (tn-368) cells. an acmnpv plaque isolate (acdefrt) was identified during propagation of a virus deficient in p35 in tn-368 cells. this virus exhibited enhanced budded-particle formation in tn-368 cells, but was partially defective for polyhedra production in the same cells. vir ...200616476974
in vivo production, stabilization, and infectivity of baculovirus preoccluded virions.wild-type and polyhedrin-negative isolates of autographa californica nuclear polyhedrosis virus were replicated in fifth-instar trichoplusia ni larvae. insect tissues infected with wild-type virus contained two types of virions that are highly infectious when ingested, those occluded in polyhedra and preoccluded virions. tissue infected with the polyhedrin-negative virus contained only preoccluded virions. the relative potencies of the two types of infected tissue were determined by dose-mortali ...199616535202
internal initiation of mrna translation in insect cell mediated by an internal ribosome entry site (ires) from shrimp white spot syndrome virus (wssv).internal initiation of mrna translation can be mediated by internal ribosome entry site (ires) elements which are located mainly in rna viruses as well as certain mammalian and insect mrna molecules. thus far, only one dna virus has been discovered to contain ires element. in this investigation, an ires element from white spot syndrome virus (wssv), a dna virus of marine shrimp, was demonstrated to direct the efficient translation of dicistronic mrna in trichoplusia ni insect cells. the ires was ...200616631622
enhancing insecticidal efficacy of baculovirus by early expressing an insect neurotoxin, lqhit2, in infected trichoplusia ni larvae.lqhit(2), an insect specific neurotoxin from the venom of leiurus quinquestriatus hebraeus, has been demonstrated to improve insecticidal efficacy of autographa californica nuclear polyhedrosis virus (acmnpv). a polyhedrin-positive recombinant acmnpvvacp(hsp70)egfp/p(pag90)it(2) was engineered for larvae to express the enhanced green fluorescence protein (egfp) and lqhit(2) under the control of p(hsp70) and p(pag90) promoters, respectively. this would allow a visual observation of the viral infe ...200616636828
arabidopsis myrosinases tgg1 and tgg2 have redundant function in glucosinolate breakdown and insect defense.in arabidopsis and other brassicaceae, the enzyme myrosinase (beta-thioglucoside glucohydrolase, tgg) degrades glucosinolates to produce toxins that deter herbivory. a broadly applicable selection for meiotic recombination between tightly linked t-dna insertions was developed to generate arabidopsis tgg1tgg2 double mutants and study myrosinase function. glucosinolate breakdown in crushed leaves of tgg1 or tgg2 single mutants was comparable to that of wild-type, indicating redundant enzyme functi ...200616640593
highly conserved piggybac elements in noctuid species of lepidoptera.the piggybac transposable element was originally discovered in a trichoplusia ni cell line and nearly identical elements were subsequently discovered in the tephritid fly, bactrocera dorsalis. this suggested the existence of piggybac in additional insects and this study shows highly conserved, though not identical, piggybac sequences in the noctuid species heliocoverpa armigera, h. zea, and spodoptera frugiperda, as well as new piggybac sequences from the t. ni organismal genome. genomic piggyba ...200616651189
the gene controlling the quantitative trait locus epithiospecifier modifier1 alters glucosinolate hydrolysis and insect resistance in arabidopsis.glucosinolates are sulfur-rich plant secondary metabolites whose breakdown products have a wide range of biological activities in plant-herbivore and plant-pathogen interactions and anticarcinogenic properties. in arabidopsis thaliana, hydrolysis by the enzyme, myrosinase, produces bioactive nitriles, epithionitriles, or isothiocyanates depending upon the plant's genotype and the glucosinolate's structure. a major determinant of this structural specificity is the epithiospecifier locus (esp), wh ...200616679459
identification and characterization of piggybac-like elements in the genome of domesticated silkworm, bombyx mori.piggybac is a short inverted terminal repeat (itr) transposable element originally discovered in trichoplusia ni. it is currently the preferred vector of choice for enhancer trapping, gene discovery and identifying gene function in insects and mammals. many piggybac-like sequences have been found in the genomes of phylogenetically species from fungi to mammals. we have identified 98 piggybac-like sequences (bmpble1-98) from the genome data of domesticated silkworm (bombyx mori) and 17 fragments ...200616685528
physical map of the dna genome of autographa californica nuclear polyhedrosis virus.a physical map of the 88 x 10(6) dalton, circular dna genome of autographa californica nuclear polyhedrosis virus was constructed. the complete order of bamhi and xmai restriction enzyme sites was determined. the ecori and hindiii fragments were partially ordered, and their general locations, relative to the bamhi and xmai maps, were determined. alterations in the restriction endonuclease fragment patterns of natural genotypic variants of a. californica nuclear polyhedrosis virus, including tric ...197916789175
restriction maps of five autographa californica mnpv variants, trichoplusia ni mnpv, and galleria mellonella mnpv dnas with endonucleases smai, kpni, bamhi, saci, xhoi, and ecori.the restriction sites of autographa californica nuclear polyhedrosis virus (acmnpv) e2 dna were mapped for the endonucleases smai, kpni, bamhi, saci, xhoi, and ecori. the restriction maps of four other acmnpv variants, trichoplusia ni (tnmnpv), and galleria mellonella (gmmnpv) genomes were determined and compared to the endonuclease cleavage maps of acmnpv e2 dna. the viral structural polypeptides of acmnpv variants s3, e2, s1, m3, and r9 were the same when analyzed by polyacrylamide gel electro ...197916789179
application of a novel radioimmunoassay to identify baculovirus structural proteins that share interspecies antigenic determinants.immunological comparisons were made of baculovirus structural proteins by using a modification of the radioimmunological techniques described by renart et al. (proc. natl. acad. sci. u.s.a. 76: 3116-3120, 1979) and towbin et al. (proc. natl. acad. sci. u.s.a. 76: 4350-4354, 1979). viral proteins were electrophoresed in polyacrylamide gels, transferred to nitrocellulose, and incubated with viral antisera, and the antibodies were detected with (125)i-labeled staphylococcus aureus protein a. antise ...198116789210
differences in susceptibility and physiological fitness of mexican field trichoplusia ni strains exposed to bacillus thuringiensis.the use of different commercial bacillus thuringiensis (bt) products in the bajio guanajuatense area in mexico began 12 yr ago, and resistance to bt in this area has been reported for plutella xylostella (l.) the current study provides a baseline response and resistance potential to bt in field and laboratory strains of bajio trichoplusia ni (hübner). differences in susceptibility to bt among t. ni populations were observed. t. ni neonates collected in romita, guanajuato, were more susceptible t ...200616813334
secretory production system of bionanocapsules using a stably transfected insect cell line.bionanocapsules (bncs) are hollow nanoscale particles composed of l protein of the hepatitis b virus surface antigen that represent specific affinity for human hepatocytes. bncs can transfer genes and drugs into human hepatocytes efficiently and specifically. bnc can be expressed in yeast cells. in this study, we developed a new l particle production system using a stably transfected insect cell line. for this purpose, we established a host-vector system using the trichoplusia ni insect cell lin ...200616847600
biosynthesis of 10,12-dienoic fatty acids by a bifunctional delta11 desaturase in spodoptera littoralis.in the biosynthetic pathway of spodoptera littoralis sex pheromone, (e,e)-10,12-tetradecadienoic acid is produced from (z)-11-tetradecenoic acid by desaturation and concomitant migration of the precursor double bond. with the aim of identifying the enzyme involved in this biotransformation, yeast deltaelo1/deltaole mutants, which are both elongase 1 and delta9 desaturase-deficient, were transformed with the s. littoralis delta11 desaturase gene using a cu+2 inducible expression vector. the trans ...200616876706
sequence and organization of the trichoplusia ni ascovirus 2c (ascoviridae) genome.the complete trichoplusia ni ascovirus 2c (tnav-2c) genome sequence was determined. the circular genome contains 174,059 bp with 165 open reading frames (orfs) of greater than 180 bp and two major homologous regions (hrs). the genome is quite a+t rich at 64.6%. fifty-four orfs had homologues in other insect viruses, such as ascoviruses, iridoviruses, baculoviruses and entomopoxviruses; 30 orfs showed low identities with those from different parasitic protozoa and 12 orfs were unique to tnav-2c. ...200616876847
piggybac-like elements in the tobacco budworm, heliothis virescens (fabricius).we identified two different groups of piggybac-like elements (ple) in the tobacco budworm, heliothis virescens, and named them hvple1 and hvple2. an intact copy of hvple1 revealed the characteristics of ple: inverted terminal repeats, inverted subterminal repeats, and an open reading frame encoding transposase, whereas other hvple1 copies and all the hvple2 copies carried disruptive mutations in the region encoding transposase. we also identified none to two bands per genome hybridized to a prob ...200616907830
importance of direct spray and spray residue contact for infection of trichoplusia ni larvae by field applications of beauveria bassiana.commercial formulations and unformulated conidia of beauveria bassiana strain gha were applied to field-grown plants and artificially infested with trichoplusia ni (hübner) larvae to compare the relative insecticidal activity resulting from direct spray contact with insecticidal activity due to contact with dry spray residue. in general, applications to cabbage, brassica oleracea l., resulted in nearly equal mortalities when comparing insects exposed to direct spray contact with those exposed by ...200616937663
suppression of plutella xylostella and trichoplusia ni in cole crops with attracticide formulations.the three key lepidopteran pests of cole, brassica oleracea l., crops in north america are diamondback moth, plutella xylostella (l.) (lepidoptera: plutellidae); cabbage looper; trichoplusia ni (hübner) (lepidoptera: noctuidae); and imported cabbageworm, pieris rapae (l.) (lepidoptera: pieridae). two species-specific pheromone-based experimental attracticide formulations were evaluated against these pests: lastcall dbm for p. xylostella and lastcall cl for t. ni. no lastcall formulation was avai ...200616937690
optimization and validation of recombinant serological tests for african swine fever diagnosis based on detection of the p30 protein produced in trichoplusia ni larvae.we describe the validation of an enzyme-linked immunosorbent assay (elisa) and confirmatory immunoblotting assays based on a recombinant p30 protein (p30r) produced in insect larvae using a baculovirus vector. such validation included the following: (i) the scaling up and standardization of p30r production and the associated immunoassays, (ii) a broad immunological analysis using a large number of samples (a total of 672) from spain and different african locations, and (iii) the detection of the ...200616954235
delta11 desaturases of trichoplusia ni and spodoptera littoralis exhibit dual catalytic behaviour.the delta(11) desaturases found in moths such as spodoptera littoralis play a critical role in the biosynthesis of their sex pheromones. the ability to functionally express these enzymes in yeast has allowed one to study the transformation of long-chain fatty acyl substrates to their 11-ene products in greater mechanistic detail. in this article, we report on the detection and quantitation of a minor 11-hydroxylated byproduct (0.1% of total fatty acids), which is formed by the delta(11) desatura ...200617027848
molecular cloning and mrna expression of peptidoglycan recognition protein (pgrp) gene in bay scallop (argopecten irradians, lamarck 1819).peptidoglycan recognition proteins (pgrps) are a type of pattern recognition molecules (prm) that recognize the unique cell wall component peptidoglycan (pgn) of bacteria and are involved in innate immunity. the first bivalve pgrp cdna sequence was cloned from bay scallop argopecten irradians by expressed sequence tag (est) and pcr technique. the full-length cdna of bay scallop pgrp (designated aipgrp) gene contained 1018bp with a 615-bp open reading frame that encoded a polypeptide of 205 amino ...200717064771
characterization of chimeric bacillus thuringiensis vip3 toxins.bacillus thuringiensis vegetative insecticidal proteins (vip) are potential alternatives for b. thuringiensis endotoxins that are currently utilized in commercial transgenic insect-resistant crops. screening a large number of b. thuringiensis isolates resulted in the cloning of vip3ac1. vip3ac1 showed high insecticidal activity against the fall armyworm spodoptera frugiperda and the cotton bollworm helicoverpa zea but very low activity against the silkworm bombyx mori. the host specificity of th ...200717122403
cell-free protein synthesis system prepared from insect cells by freeze-thawing.we established a novel cell-free protein synthesis system derived from trichoplusia ni (highfive) insect cells by a simple extraction method. luciferase and beta-galactosidase were synthesized in this system with active forms. we analyzed and optimized (1) the preparation method of the insect cell extract, (2) the concentration of the reaction components, and (3) the 5'-untranslated region (5'-utr) of mrna. the extract was prepared by freeze-thawing insect cells suspended in the extraction buffe ...200617137303
functional diversification of acyl-coenzyme a oxidases in jasmonic acid biosynthesis and action.the biosynthesis of jasmonic acid (ja) in plant peroxisomes requires the action of acyl-coenzyme a oxidase (acx). among the five expressed members (acx1-5) of the acx gene family in arabidopsis (arabidopsis thaliana), only acx1 is known to serve a role in ja production. here, we used transgenic promoter-reporter lines to show that acx1 is highly expressed in mature and germinating pollen, stem epidermal cells, and other tissues in which jasmonate-signaled processes occur. wound-induced ja accumu ...200717172287
infection, transfection, and co-transfection of baculoviruses by microprojectile bombardment of larvae.the use of baculoviruses as expression vectors for heterologous proteins has been practically limited to the use of the autographa californica multiple nucleopolyhedrovirus (acmnpv). in this work, infection, transfection and co-transfection events with the baculoviruses acmnpv and trichoplusia ni granulovirus (tngv) were accomplished by bombardment of t. ni first-instar larvae with microprojectiles coated with virions, viral dna, and viral dna and a transfer vector, respectively. a series of sho ...200717184851
mechanism of resistance to bacillus thuringiensis toxin cry1ac in a greenhouse population of the cabbage looper, trichoplusia ni.the cabbage looper, trichoplusia ni, is one of only two insect species that have evolved resistance to bacillus thuringiensis in agricultural situations. the trait of resistance to b. thuringiensis toxin cry1ac from a greenhouse-evolved resistant population of t. ni was introgressed into a highly inbred susceptible laboratory strain. the resulting introgression strain, glen-cry1ac-bcs, and its nearly isogenic susceptible strain were subjected to comparative genetic and biochemical studies to det ...200717189446
host-plant effects the expression of resistance to bacillus thuringiensis kurstaki in trichoplusia ni (hubner): an important factor in resistance evolution.pathogens are thought to exert strong selection on their hosts leading to increased host resistance. bacillus thuringiensis kurstaki (bkt) is a ubiquitous entomopathogen that has become the mainstay of nonchemical control of lepidopteran pests and thus, the potential exists for the evolution of resistance in targeted host insects. we have studied the expression of btk resistance in the cabbage looper, trichoplusia ni (hubner). for this generalist insect herbivore, three common host plants, tomat ...200717210000
expression, purification, and characterization of human acetyl-coa carboxylase 2.the full-length human acetyl-coa carboxylase 1 (acc1) was expressed and purified to homogeneity by two separate groups (y.g. gu, m. weitzberg, r.f. clark, x. xu, q. li, t. zhang, t.m. hansen, g. liu, z. xin, x. wang, t. mcnally, h. camp, b.a. beutel, h.i. sham, synthesis and structure-activity relationships of n-{3-[2-(4-alkoxyphenoxy)thiazol-5-yl]-1-methylprop-2-ynyl}carboxy derivatives as selective acetyl-coa carboxylase 2 inhibitors, j. med. chem. 49 (2006) 3770-3773; d. cheng, c.h. chu, l. c ...200717223360
metabolic engineering of the baculovirus-expression system via inverse "shotgun" genomic analysis and rna interference (dsrna) increases product yield and cell longevity.rna interference (rnai) is as powerful tool for characterizing gene function in eukaryotic organisms and cultured cell lines. its use in metabolic engineering has been limited and few reports have targeted protein expression systems to increase yield. in this work, we examine the use of in vitro synthesized double stranded rna (dsrna) in the baculovirus expression vector system (bevs), using commercially relevant cultured cells (spodoptera frugiperda, sf-9) and larvae (trichoplusia ni) as hosts. ...200717274062
development of a rapid chromatographic immunoassay for detection of human metapneumovirus using monoclonal antibodies against nucleoprotein of hmpv.human metapneumovirus (hmpv) nucleocapsid (n) protein is a major structural protein that encapsidates the rna genome and is essential for replication and transcription of the hmpv genome. we developed two mouse monoclonal antibodies (mabs), designated 3d1 and 5b10, against n protein of hmpv and characterized them by an immunofluorescence assay and an immunoprecipitation assay using trichoplusia ni (tn5) insect cells infected with a recombinant baculovirus-expressing hmpv n protein. both mabs wer ...200717316081
increased plasma selenium levels correlate with elevated resistance of heliothis virescens larvae against baculovirus infection.we reported that dietary selenium (se) impacted the growth and development of trichoplusia ni reared for many generations on diet containing extremely low levels of se. larvae had an elevated resistance to per os infection with a baculovirus. in this study, we examine how dietary se (in the form of selenite) affects the growth, development, and se content of heliothis virescens that have been laboratory reared for less than two years. larvae fed a commercial tobacco budworm diet supplemented wit ...200717316679
sequence and organization of the heliothis virescens ascovirus genome.the nucleotide sequence of the heliothis virescens ascovirus (hvav-3e) dna genome was determined and characterized in this study. the circular genome consists of 186,262 bp, has a g+c content of 45.8 mol% and encodes 180 potential open reading frames (orfs). five unique homologous regions (hrs), 23 'baculovirus repeat orfs' (bro) and genes encoding a caspase homologue and several enzymes involved in nucleotide replication and metabolism were found in the genome. several ascovirus (av)-, iridovir ...200717374755
stability of plant defense proteins in the gut of insect herbivores.plant defense against insect herbivores is mediated in part by enzymes that impair digestive processes in the insect gut. little is known about the evolutionary origins of these enzymes, their distribution in the plant kingdom, or the mechanisms by which they act in the protease-rich environment of the animal digestive tract. one example of such an enzyme is threonine (thr) deaminase (td), which in tomato (solanum lycopersicum) serves a dual role in isoleucine (ile) biosynthesis in planta and th ...200717416643
effect of bt cotton expressing cry1ac and cry2ab, non-bt cotton and starvation on survival and development of trichoplusia ni (lepidoptera: noctuidae).effects of bollgard ii cotton containing two bacillus thuringensis var. kurstaki berliner (bt) toxin proteins (cry1ac and cry2ab), non-bt cotton (dpl 491) and starvation on survival and development of cabbage looper, trichoplusia ni (hübner), were determined in the laboratory. larvae of the first four larval instars died when they fed on the terminal leaves of bt cotton plants at 50 days after planting (dap). however, 51.3% of fifth instars that fed on 50 dap bt cotton leaves pupated, and 87.1% ...200717421053
development of a low-cost, insect larvae-derived recombinant subunit vaccine against rhdv.vaccine antigens against rabbit hemorrhagic disease virus (rhdv) are currently derived from inactivated rhdv obtained from livers of experimentally infected rabbits. several rhdv-derived recombinant immunogens have been reported. however, their application in vaccines has been restricted due to their high production costs. in this paper, we describe the development of an inexpensive, safe, stable vaccine antigen for rhdv. a baculovirus expressing a recombinant rhdv capsid protein (vp60r) was use ...200717434554
the autographa californica m nucleopolyhedrovirus fibroblast growth factor accelerates host mortality.the fibroblast growth factor (vfgf) gene encoded by autographa californica m nucleopolyhedrovirus (acmnpv) has been shown to share functional properties with cellular fgfs; it is a secreted protein, binds heparin, and stimulates motility of insect cells. we previously reported that viruses containing or lacking vfgf produced similar yields of budded virus and had similar kinetics of viral dna and protein syntheses in cultured cells. in this study, we characterized these viruses in two permissive ...200717459443
propagation of bombyx mori nucleopolyhedrovirus in nonpermissive insect cell lines.this study addresses the susceptibility of spodoptera frugiperda (sf9 and sf21), trichoplusia ni (hi5), and s. exigua (se301) cells to the bombyx mori nucleopolyhedrovirus (bmnpv). although these cells have classically been considered nonpermissive to bmnpv, the cytopathic effect, an increase in viral yield, and viral dna synthesis by bmnpv were observed in sf9, sf21, and hi5 cells, but not in se301 cells. very late gene expression by bmnpv in these cell lines was also detected via beta-galactos ...200717483798
characterisation and partial sequence analysis of two novel cypoviruses isolated from the winter moth operophtera brumata (lepidoptera: geometridae).the complete nucleotide sequences of genomic segments s5 to s10 from operophtera brumata cypovirus 18 (opbucpv18), and the complete nucleotide sequences of genomic segments s2, s5, s9 and s10 from operophtera brumata cypovirus 19 (opbucpv19) have been determined. each genome segment contained a single open reading frame (orf). conserved motifs 5' (aguaaa....guuagcu) 3' were found at the ends of each segment of opbucpv18, whilst conserved motifs 5' (aacaaa....uuugc) 3' were found at each segment ...200717546493
ac18 is not essential for the propagation of autographa californica multiple nucleopolyhedrovirus.orf18 (ac18) of autographa californica multiple nucleopolyhedrovirus (acmnpv) is a highly conserved gene in lepidopteran nucleopolyhedroviruses, but its function remains unknown. in this study, an ac18 knockout acmnpv bacmid was generated to determine the role of ac18 in baculovirus life cycle. after transfection of sf-9 cells, the ac18-null mutant showed similar infection pattern to the parent virus and the ac18 repair virus with respect to the production of infectious budded virus, occlusion b ...200717573091
piggybac transposon mediated transgenesis of the human blood fluke, schistosoma mansoni.the transposon piggybac from the genome of the cabbage looper moth trichoplusia ni has been observed in the laboratory to jump into the genomes of key model and pathogenic eukaryote organisms including mosquitoes, planarians, human and other mammalian cells, and the malaria parasite plasmodium falciparum. introduction of exogenous transposons into schistosomes has not been reported but transposon-mediated transgenesis of schistosomes might supersede current methods for functional genomics of thi ...200717586730
production of selenomethionyl-derivatized proteins in baculovirus-infected insect cells.a protocol is described for the production of both intracellularly expressed and secreted selenomethionyl-derivatized recombinant proteins in baculovirus-infected insect cells. the method results in the production of recombinant soluble proteins with an semet occupancy of approximately 75% and with a recovery of approximately 20% that of native protein expression. the method is independent of the percentage methionine content of the protein and is reliable and consistent. similar results are obt ...200717660253
functional comparison of full-length and n-terminal-truncated octopamine transporters from lepidoptera.we have cloned two new lepidopteran octopamine transporters (oats), members of the solute-linked carrier family 6 (slc6) of nutrient transporters, from the cns of the european corn borer ostrinia nubilalis and the cabbage white pieris rapae. comparison of these sequences with the previously cloned oat from the cabbage looper trichoplusia ni showed that the t. ni oat sequence previously reported was truncated by 74 amino acids at the n-terminus. the cytoplasmic n-termini deduced here are consider ...200717681232
seroepidemiology of human bocavirus in hokkaido prefecture, japan.a new human virus, provisionally named human bocavirus (hbov), was discovered by swedish researchers in 2005. a new immunofluorescence assay using trichoplusia ni insect cells infected with a recombinant baculovirus expressing the vp1 protein of hbov was developed, and the levels of immunoglobulin g antibody to the vp1 protein of hbov in serum samples were measured. the overall seroprevalence rate of antibodies against the vp1 protein of hbov in a japanese population aged from 0 months to 41 yea ...200717699639
production of a recombinant antibody fragment in whole insect larvae.infection of insect cells with baculovirus expression constructs is commonly used to produce recombinant proteins that require post-translational modifications for their activity, such as mammalian proteins. however, technical restraints limit the capacity of insect cell-based culture systems to be scaled up to produce the large amounts of recombinant protein required for human pharmaceuticals. in this study, we designed an automated insect rearing system and whole insect baculovirus expression ...200717827537
purification and characterization of native and recombinant sapin2a, a plant sieve element-localized proteinase inhibitor.sapin2a encodes a proteinase inhibitor in nightshade (solanum americanum), which is specifically localized to the enucleate sieve elements. it has been proposed to play an important role in phloem development by regulating proteolysis in sieve elements. in this study, we purified and characterized native sapin2a from nightshade stems and recombinant sapin2a expressed in escherichia coli. purified native sapin2a was found as a charge isomer family of homodimers, and was weakly glycosylated. nativ ...200717870592
development of serum-free media for lepidopteran insect cell lines.lepidopteran insect cell culture technology has progressed to the point of becoming an essential part of one of the most successful eukaryotic expression systems and is increasingly used industrially on a large scale. therefore, there is a constant need for convenient and low-cost culture media capable of supporting good insect cell growth and ensuring high yield of baculovirus as well as the strong expression of recombinant proteins. vertebrate sera or invertebrate hemolymph were essential supp ...200717951770
baculovirus insecticide production in insect larvae.baculovirus-based insecticides are currently being used worldwide, and new products are in development in many countries. the most dramatic examples of successful baculovirus insecticides are found in soybean in brazil and cotton in china. production of baculoviruses is generally done in larvae of a convenient host species, and the level of sophistication varies tremendously between field-collection of infected insects at the one extreme and automated mass manufacturing at the other. currently, ...200717951781
adaptation of the "in-gel release method" to n-glycome analysis of low-milligram amounts of material.protein n-glycosylation is a post-translational modification which plays numerous crucial physiological roles. the n-glycan pattern varies depending on the species organs, tissues and even cell types and their respective physiological states. obtaining enough starting material from a particular cell type or tissue for n-glycan purification by conventional methods can, in certain cases, be very difficult. previously, a sensitive technique, the "in-gel release method" that allows the determination ...200718041037
production of recombinant polyhedra containing cry1ac fusion protein in insect cell lines.insect cell lines and the control of infection for obtaining the maximum amount of polyhedrin-crylac-polyhedrin fusion protein from bactrus in monolayer and suspension culture systems were tested. growth rates of the trichoplusia ni (high-five) cell line in both culture systems were better than the other insect cell lines, spodoptera fiugiferda (sf-9, sf-21), trichoplusia ni (tn5), and spodoptera exigua (se301). the expression of the fusion protein in a monolayer culture showed that se301 cells ...200718051294
novel isolate of bacillus thuringiensis subsp. thuringiensis that produces a quasicuboidal crystal of cry1ab21 toxic to larvae of trichoplusia ni.a new isolate (is5056) of bacillus thuringiensis subsp. thuringiensis that produces a novel variant of cry1ab, cry1ab21, was isolated from soil collected in northeastern poland. cry1ab21 was composed of 1,155 amino acids and had a molecular mass of 130.5 kda, and a single copy of the gene coding for this endotoxin was located on a approximately 75-kbp plasmid. when synthesized by the wild-type strain, cry1ab21 produced a unique, irregular, bipyramidal crystal whose long and short axes were both ...200818083867
immune system responses and fitness costs associated with consumption of bacteria in larvae of trichoplusia ni.insects helped pioneer, and persist as model organisms for, the study of specific aspects of immunity. although they lack an adaptive immune system, insects possess an innate immune system that recognizes and destroys intruding microorganisms. its operation under natural conditions has not been well studied, as most studies have introduced microbes to laboratory-reared insects via artificial mechanical wounding. one of the most common routes of natural exposure and infection, however, is via foo ...200718154650
positive association between resistance to bacillus thuringiensis and overwintering survival of cabbage loopers, trichoplusia ni (lepidoptera: noctuidae).development of resistance to insecticides has generally been associated with fitness costs that may be magnified under challenging conditions. lepidopterans which are resistant to the biopesticide bacillus thuringiensis subsp. kurstaki (btk) have been shown to have reduced fitness, such as lower survival when subjected to overwintering stress. recently, resistance to btk has been found in some populations of trichoplusia ni hübner in greenhouses in british columbia. this situation provides an op ...200818257958
extended budded virus formation and induction of apoptosis by an acmnpv fp-25/p35 double mutant in trichoplusia ni cells.an autographa californica nucleopolyhedrovirus (acmnpv) mutant (acdefrt) isolated from virus-infected trichoplusia ni (tn-368) cells produced plasma membrane blebbing and caspase-3-like activity late in infection. it also synthesized less polyhedra, but displayed enhanced budded virus formation in tn-368 cells. this phenotype resulted from dual mutations in p35 and fp-25. in this study we showed that enhanced budded virus production occurs because the hourly rate of release of virus from acdefrt ...200818261819
expression and characterization of human n-acetylglucosaminyltransferases and alpha2,3-sialyltransferase in insect cells for in vitro glycosylation of recombinant erythropoietin.the glycans linked to the insect cell-derived glycoproteins are known to differ from those expressed mammalian cells, partly because of the low level or lack of glycosyltransferase activities. gnt ii, gnt iv, gnt v, and st3gal iv, which play important roles in the synthesis of tetraantennary-type complex glycan structures in mammalian cells, were overexpressed in trichoplusia ni cells by using a baculovirus expression vector. the glycosyltransferases, expressed as a fusion form with the igg-bind ...200818309288
detection of genes encoding antimicrobial peptides in mexican strains of trichoplusia ni (hübner) exposed to bacillus thuringiensis.the systemic immune response of trichoplusia ni after bacillus thuringiensis (bt) exposure was evaluated by comparing the expression of genes encoding antimicrobial peptides (amps) in bt-susceptible and -resistant t. ni strains that were either exposed or not to xentari (bt-xt). amp genes were detected by rt-pcr using primers for attacin, gloverin, lebocin, lysozyme, and peptidoglycan recognition peptide (pgrp). in general, amp genes were detected more frequently in mexican field strains previou ...200818359041
diversity of errantivirus (retrovirus) sequences in two cell lines used for baculovirus expression, spodoptera frugiperda and trichoplusia ni.to determine the diversity of retroviruses (errantiviruses) in cell lines used for baculovirus expression, degenerate primers were designed complementary to conserved regions of lepidopteran errantivirus reverse transcriptase genes. these primers were used to pcr amplify sequences from dna isolated from spodoptera frugiperda (sf-9) and trichoplusia ni (hi-5) cell lines. cloning, sequencing, and phylogenetic analysis of over 20 pcr products from each cell line demonstrated the presence of diverse ...200818363091
plant-mediated alteration of the peritrophic matrix and baculovirus infection in lepidopteran larvae.the peritrophic matrix (pm) lines the midgut of most insects, providing protection to the midgut epithelial cells while permitting passage of nutrients and water. herein, we provide evidence that plant-mediated alteration of the pm contributes to the well-documented inhibition of fatal infection by autographa californica multiple nucleopolyhedrovirus (acmnpv) of heliothis virescens f. larvae fed cotton foliage. we examined the impact of the pm on pathogenesis using a viral construct expressing a ...200818374352
the iridoid glucoside, antirrhinoside, from antirrhinum majus l. has differential effects on two generalist insect herbivores.the iridoid glucoside, antirrhinoside, is constitutively distributed throughout antirrhinum majus l. in a manner consistent with its possible role as an allelochemical, but there is no evidence that it has a defensive function with respect to insect herbivory. to address this question, two generalist herbivores, lymantria dispar l. (gypsy moth) and trichoplusia ni hübner (cabbage looper) were chosen for feeding trials on excised whole leaves of a. majus and in artificial diet assays. in leaf exc ...200818414950
characterization of acmnpv with a deletion of ac68 gene.orf68 (ac68) of autographa californica multiple nucleopolyhedrovirus (acmnpv) is a highly conserved gene that codes a predicted 192-amino acid protein, but its function remains unknown. results of the current study showed that ac68 was transcribed from 3 to 96 h and the protein was detected from 36 to 96 h post infection. an ac68 knockout bacmid was generated to investigate the role of the gene in baculovirus life cycle. analyses of the production of infectious budded virus, occlusion bodies, an ...200818483845
expression and production of human interleukin-7 in insect cells using baculovirus expression vector system (bevs).interleukin-7 (il-7) is a glycoprotein cytokine with significant clinical and biomedical potential, such as cancer therapy and hiv infections. earlier it has been cloned and expressed in various protein expression systems; however, they are not efficient for large-scale production. to address this inadequacy, we report in this paper the production of recombinant human interleukin-7 (hil-7) in insect cells. a recombinant bacmid containing hil-7 was constructed, purified, and characterized. it was ...200818546077
genotoxic activation of 2-phenylbenzotriazole-type compounds by human cytochrome p4501a1 and n-acetyltransferase expressed in salmonella typhimurium umu strains.four 2-phenylbenzotriazole (pbta)-type compounds (pbta-4, pbta-6, pbta-7, and pbta-8) were identified as major mutagens in blue cotton/rayon-adsorbed substances collected at sites below textile dyeing factories or municipal water treatment plants treating domestic waste and effluents from textile dyeing factories in several rivers in japan. the main purpose of this study is to understand the basis of the roles of human cytochrome p450 (cyp) and n-acetyltransferases (nats) in genotoxic activation ...200818562244
genomic organization of the translationally controlled tumor protein (tctp) gene from shrimp marsupenaeus japonicus.the shrimp translationally controlled tumor protein (tctp) gene was previously found up-regulated in response to viral infection. in order to further understand the transcriptional regulation of tctp gene expression under viral challenge, the 5'-flanking promoter region of the tctp gene in shrimp marsupenaeus japonicus (mj-tctp) was cloned by using genomic walking method. the sequence shows that this region contains many important potential binding sites for transcription factors, such as activa ...200918587664
a new trichoplusia ni cell line for membrane protein expression using a baculovirus expression vector system.a new cell line, msu-tnt4 (tnt4), was established from trichoplusia ni embryos for use with baculovirus expression vectors and evaluated for its potential for membrane protein production. to evaluate membrane protein synthesis, recombinant baculoviruses were constructed to express the human neurotensin receptor 1 as an enhanced green fluorescent protein (gfp) fusion. tnt4 cells had a doubling time of 21 h and expressed the membrane-gfp fusion protein at approximately twice the level as sf21 cell ...200818622664
rational scale-up of a baculovirus-insect cell batch process based on medium nutritional depth.we have developed a serum-free cell culture process utilizing a recombinant baculovirus (acnpv) expression vector to infect trichoplusia ni insect cells for the production of the human lysosomal enzyme, glucocerebrosidase. the enzyme, which is harvested as a secreted protein in this process, can serve as a replacement therapy for the genetic deficiency gaucher disease. in the course of pilot scale-up of a batch glucocerebrosidase process from 25-ml working volume shaker flask units to 25-l worki ...199618629948
autographa californica nuclear polyhedrosis virus-induced proteins in tissue culture.the multiplication of autographa californica nuclear polyhedrosis virus was studied in trichoplusia ni and spodoptera frugiperda insect cell lines. using pulse labeling, 18 viral-induced proteins were identified by polyacrylamide gel electrophoresis. based on infectivity curves and protein synthesis studies, replication could be divided into four phases. during the early phase of infection, four viral-induced proteins with molecular weights of 45, 35, 34, and 31 thousand daltons could be detecte ...198018631644
radioimmunoassay analysis of baculovirus granulins and polyhedrins.granulin and polyhedrin proteins were purified by preparative sodium dodecyl sulfate (sds)-polyacrylamide gel electrophoresis from the baculoviruses autographa californica, rachiplusia ou, heliothis zea, heliothis armigera, trichoplusia ni, and spodoptera frugiperda. antisera were raised against autographa californica (ac) polyhedrin and trichoplusia ni (tn) granulin and analyzed for homologous and heterologous immunoreactivity by immunodiffusion and radioimmunoassay (ria). ac polyhedrin and tn ...198018631650
isolation and replication of an occlusion body-deficient mutant of the autographa californica nuclear polyhedrosis virus.a spontaneous mutant of the autographa californica nuclear polyhedrosis virus which was deficient in viral occlusion body formation was studied in trichoplusia ni tissue culture cells. virus multiplication could be divided into four phases based on growth curves and the sequential appearance of 25 viral-induced proteins identified by pulse labeling and polyacrylamide gel electrophoresis. during the latent phase, five viral-induced proteins with molecular weights of 68, 47, 35, and 30 thousand (k ...198018631676
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