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modification of trichoplusia ni nuclear polyhedrosis virus passaged in vivo.a designated strain of plaque-purified trichoplusia ni nuclear polyhedrosis virus (npv) was used to initiate a serial passage series in vivo. the virulence of polyhedra inclusion bodies (pib) and the distribution of mp (many pib) and fp (few pib) strains in hemolymph were monitored. when virus was passaged per os as pib, there was no major change in virulence after 15 passes in insects. plaque analysis of hemolymph from the infected insects indicated that there was no selection for the fp strain ...1978338547
restriction endonuclease analysis to distinguish two closely related nuclear polyhedrosis viruses: autographa californica mnpv and trichoplusia ni mnpv.restriction endonuclease fragment patterns of the deoxyribonucleic acid genomes of autographa californica nuclear polyhedrosis virus (multiply embedded type) and trichoplusia ni nuclear polyhedrosis virus (multiply embedded type) demonstrate that the two viruses are distinct but closely related variants.1978354530
interference with polyhedral inclusion body (pib) production in trichoplusia ni cells infected with a high passage strain of autographa californica nuclear polyhedrosis virus (npv). brief report.trichoplusia ni cells infected with a low passage (lp) strain of autographa californica nuclear polyhedrosis virus (npv) produce large numbers of polyhedral inclusion bodies (pibs). interference with pib production occurs when t. ni cells are first inoculated with a high passage (hp) strain and then challenged with the lp strain. pib production is reduced 100 fold to the level seen with hp virus infection only.1979389204
pathogenicity of bacillus cereus isolated from trichoplusia ni larvae. 1975807657
effect of a nuclear polyhedrosis virus on the realtionship between trichoplusia ni (lepidoptera: noctuidae) and the parasite, hyposoter exiguae (hymenoptera: ichneumonidae). 19751089733
degradation of matrix protein from a nuclear-polyhedrosis virus of trichoplusia ni by an endogenous protease. 19751103444
bacterial luciferase produced with rapid-screening baculovirus vectors is a sensitive reporter for infection of insect cells and larvae.bacterial luciferase, derived from a fusion of the luxa and luxb genes of vibrio harveyi, has been expressed at very high levels in caterpillars and insect cells. the coding sequence for luciferase was inserted into vectors developed in our laboratory which were designed to expedite screening of recombinant virus. these vectors contained the beta-galactosidase indicator gene under control of immediate early (ie1), early (etl), or very late (p10) promoters and a cloning site for inserting the fus ...19921308004
microscopic visualization of insect cell-bubble interactions. ii: the bubble film and bubble rupture.in this paper, the second in the series, the use of a microscopic, high-speed video system to study the interactions of two suspended insect cells strains, trichoplusia ni (tn-368) and spodoptera frugiperda (sf-9), with rupturing bubbles is reported. events such as the adsorption of cells onto the bubble film and the mechanism of bubble rupture were observed. on the basis of these observations and the experimental and theoretical work of other researchers on bubble rupture and cell death as a re ...19911367170
improved efficiency in determining the titer of the autographa californica baculovirus nonoccluded virus.an economical and time-efficient method for titering the autographa californica multiple-embedded nuclear polyhedrosis virus (acmnpv) by the endpoint method is described. the method uses an electronic pipetting device to perform dilutions in the same 60-well microplate as is used for the assay, thus eliminating the need for test tubes or vials for making dilutions. additionally, since small volumes are used for the dilutions, a substantial savings in medium is achieved. the effect of using three ...19921389161
use of a colorimetric system to detect enzymes expressed by germinating conidia of entomopathogenic fungi.an apizym system, with 19 substrates, was used to detect enzymes expressed by germinating conidia of nomuraea rileyi (5 isolates), nomuraea atypicola, nomuraea anemonoides, beauveria bassiana and metarhizium anisopliae. similar enzyme profiles were obtained for two of the n. rileyi isolates (mississippi, ecuador) regardless of whether culture medium (sabouraud-maltose-yeast) or cuticle (from larvae of trichoplusia ni, heliothis zea or heliothis virescens) were used as substrates. centroid-cluste ...19921406899
site-specific mutagenesis of the 35-kilodalton protein gene encoded by autographa californica nuclear polyhedrosis virus: cell line-specific effects on virus replication.the gene encoding the 35-kda protein (35k gene) located within the ecori-s genome fragment of autographa californica nuclear polyhedrosis virus (acmnpv) is transcribed early in infection. to examine its function(s) with respect to virus multiplication, we introduced specific mutations of this early gene into the acmnpv genome. in spodoptera frugiperda (sf21) culture, deletion of the 35k gene reduced yields of extracellular, budded virus from 200- to 15,000-fold, depending on input multiplicity. ...19921501287
in vivo transcriptional analysis of three baculovirus genes: evidence of homology between viral and host transcripts.transcripts for the gp64 and polyhedrin genes as well as a 1629 open reading frame (1629 orf) of the autographa californica nuclear polyhedrosis virus were examined in the midgut tissues and hemocytes of uninfected and infected host trichoplusia ni larvae and sf21 cells. polyhedrin-specific transcripts of 1.2, 3.4, and 4.9 kb were expressed in both infected larval tissues and infected sf21 cells. the highest level of expression for polyhedrin-specific transcripts was observed in hemocytes, where ...19921529534
comparative toxicity of the hd-1 and nrd-12 strains of bacillus thuringiensis subsp. kurstaki to defoliating forest lepidoptera.insecticidal activities of sporulated cultures of the hd-1 and nrd-12 strains of bacillus thuringiensis subsp. kurstaki were compared against four species of defoliating forest lepidopterans in diet-incorporation assays. there was no difference in lc50 between the two strains to larvae of spruce budworm (choristoneura fumiferana), gypsy moth (lymantria dispar), eastern hemlock looper (lambdina fiscellaria fiscellaria), and whitemarked tussock moth (orgyia leucostigma), whether expressed as total ...19921607666
biologically active and amidated cecropin produced in a baculovirus expression system from a fusion construct containing the antibody-binding part of protein a.a synthetic antibody-binding part derived from protein a from staphylococcus aureus was used as a fusion partner in a eukaryotic expression system employing autographa californica nuclear polyhedrosis as a vector. this, in conjunction with an efficient signal sequence, facilitated the purification of the antibacterial peptide cecropin a from the medium of spodoptera frugiperda cells infected with a recombinant virus. in order to increase further the concentrations of fusion protein, trichoplusia ...19911720614
expression of hepatitis b virus surface antigen gene in insect baculovirus vector systems.the gene coding for the hepatitis b virus surface antigen (hbsag) under the control of autographa californicanuclear polyhedrosis virus polyhedrin promoter was successfully inserted into the genome of the trichoplusia ni nuclear polyhdrosis virus. infection of spodoptera frugiperda cells with this recombinant virus produced a significant amount of hbsag protein and secreted 22 nm particles containing the hbsag. the expression of hbsag gene was also obtained both in trichoplusia ni larvae and in ...19911773016
location and nucleotide sequence of the gene encoding the viral enhancing factor of the trichoplusia ni granulosis virus.the gene encoding the viral enhancing factor (vef) of trichoplusia ni granulosis virus has been cloned from a lambda gt11 expression library, and the complete nucleotide sequence determined. the vef gene encodes a protein with a predicted mr of 104k which does not share homology with any previously reported proteins. a possible promoter is located four nucleotides upstream of the initiation codon and represents a consensus baculovirus late promoter (ataag). this has been confirmed by the identif ...19911940861
development of mutants of the mosquitocidal bacterium bacillus thuringiensis subspecies morrisoni (pg-14) toxic to lepidopterous or dipterous insects.the parasporal body of the mosquitocidal isolate (pg-14) of bacillus thuringiensis subsp. morrisoni (btm) contains five major proteins with molecular masses of, respectively, 27.3, 65, 128, 135, and 144 kda. proteins corresponding in mass to the first four of these also occur in the mosquitocidal strain, b. thuringiensis subsp. israelensis (bti), and it is thought therefore that the mosquitocidal activity of both strains is due to these four proteins. in other studies it has been shown that each ...19901969828
chemical modification of bacillus thuringiensis subsp. thuringiensis (hd-524) trypsin-activated endotoxin: implication of tyrosine residues in lepidopteran cell lysis.a purified protein fraction from a solubilized and trypsin-digested extract of bacillus thuringiensis subsp. thuringiensis (hd-524) fermentation powder was lytic to cells from several lepidopteran lines. maximum yield was obtained by alkaline carbonate-thiocyanate solubilization of washed powder followed by trypsin digestion and sephacryl (s-300) chromatography. the alkaline carbonate-solubilized fraction consisted predominantly of two bands on sodium dodecyl sulfate-polyacrylamide gel electroph ...19912002242
insecticidal toxins from bacillus thuringiensis subsp. kenyae: gene cloning and characterization and comparison with b. thuringiensis subsp. kurstaki cryia(c) toxins.genes encoding insecticidal crystal proteins were cloned from three strains of bacillus thuringiensis subsp. kenyae and two strains of b. thuringiensis subsp. kurstaki. characterization of the b. thuringiensis subsp. kenyae toxin genes showed that they are most closely related to cryia(c) from b. thuringiensis subsp. kurstaki. the cloned genes were introduced into bacillus host strains, and the spectra of insecticidal activities of each cry protein were determined for six pest lepidopteran insec ...19912014985
activation of a cryptic crystal protein gene of bacillus thuringiensis subspecies kurstaki by gene fusion and determination of the crystal protein insecticidal specificity.dna hybridization with the insecticidal crystal protein gene crylla (formerly crybl) of bacillus thuringiensis supspecies kurstaki has shown that subspecies kurstaki contains a crylla-related sequence in addition to the crylla gene (donovan et al., 1988a). we have cloned the crylla-related sequence and have determined that the sequence, which has been designated cryllb, is 89% identical to the crylla gene. recombinant b. thuringiensis cells harbouring the cloned cryllb gene produced very little ...19902089222
molecular characterization of immune inhibitor a, a secreted virulence protease from bacillus thuringiensis.the gene for the secreted neutral metalloprotease, immune inhibitor a (ina), from bacillus thuringiensis var. alesti has been cloned and sequenced. the deduced amino acid sequence has been confirmed by partial amino acid sequencing. the central part of the amino acid sequence showed similarity to the active site in thermolysin. southern and western blots show that ina-related sequences are common among other b. thuringiensis subspecies. in western blots, 17 out of 25 tested species gave a positi ...19902089225
construction of genetically engineered baculovirus insecticides containing the bacillus thuringiensis subsp. kurstaki hd-73 delta endotoxin.the delta-endotoxin gene from bacillus thuringiensis subsp. kurstaki hd-73 was inserted into autographa californica nuclear polyhedrosis virus (acmnpv) using two transfer vector systems. in the first, the delta-endotoxin gene was placed under the control of the polyhedrin gene promoter in lieu of the polyhedrin coding sequences, thus deriving a polyhedrin-negative virus. in the second, it was inserted under the control of a copy of the acmnpv p10 promoter positioned upstream of the polyhedrin ge ...19902165136
mechanisms for regulating oxygen toxicity in phytophagous insects.the antioxidant enzymatic defense of insects for the regulation of oxygen toxicity was investigated. insect species examined were lepidopterous larvae of the cabbage looper (trichoplusia ni), southern armyworm (spodoptera eridania), and black swallowtail (papilio polyxenes). these phytophagous species are subject to both endogenous and exogenous sources of oxidative stress from toxic oxygen radicals, hydrogen peroxide (h2o2) and lipid peroxides (looh). in general, the constitutive levels of the ...19902199343
infected cell specific protein and viral dna synthesis in productive and abortive infections of spodoptera frugiperda nuclear polyhedrosis virus.this study examines viral protein and dna synthesis in spodoptera frugiperda multicapsid nuclear polyhedrosis virus (sfmnpv) infections of s. frugiperda and trichoplusia ni cells. a total of 28 infected cell specific polypeptides (icsps) were detected in the productive s. frugiperda cells. of these, 14 were identified as structural polypeptides. based on the change in their rate of synthesis during the replication cycle, these icsps were grouped into four classes. only a 97k and a 29k icsp were ...19902248550
comparative histopathology of three ascovirus isolates in larval noctuids.the histopathology caused by three ascoviruses isolated respectively from heliothis zea, spodoptera frugiperda, and trichoplusia ni was studied in the host species from which the viruses were isolated originally, or in the case of the h. zea isolate, in h. virescens. in all three isolates, infected cells and virion-containing vesicles in the hemolymph were observed by 3 days postinoculation. the isolates from h. zea and t. ni exhibited relatively broad tissue tropisms infecting the tracheal matr ...19902250100
comparative study of virion structure, protein composition and genomic dna of three ascovirus isolates.the virions of three ascoviruses isolated from the noctuids heliothis zea, spodoptera frugiperda and trichoplusia ni were compared with respect to their size and structure, protein composition and the size and relatedness of their dnas. the virions of the isolates from h. zea (hav) and t. ni (tav) were allantoid in shape (400 x 130 nm), enveloped and contained an inner particle which appeared to have an internal lipid bilayer surrounding the dna core. the virions of the s. frugiperda isolate (sa ...19902391498
toxicity to spodoptera exigua and trichoplusia ni of individual p1 protoxins and sporulated cultures of bacillus thuringiensis subsp. kurstaki hd-1 and nrd-12.the toxicities to neonate spodoptera exigua and trichoplusia ni of lyophilized powders obtained from sporulated liquid cultures (referred to as sporulated cultures) and escherichia coli-expressed p1 [cryia(a) cryia(b) cryia(c)] protoxins from three-gene strains of nrd-12 and hd-1 of bacillus thuringiensis subsp. kurstaki were determined by using diet incorporation bioassays. although sporulated cultures from both strains were more toxic to t. ni than s. exigua, there were no differences in toxic ...19902403254
microscopic visualization of insect cell-bubble interactions. i: rising bubbles, air-medium interface, and the foam layer.through the use of microscopic, high-speed video technology, the interactions of two suspended insect cell lines, trichoplusia ni (tn-368) and spodoptera frugiperda (sf-9), with air and oxygen bubbles were studied. events such as cell-bubble attachment, cell-bubble collision, cell transport into the foam layer, and trapping of cells in the foam layer are presented and discussed. based on these observations and those in a companion paper (chalmers, j. j.; bavarian, f. biotechnol. prog. 1991, foll ...19911367169
structure of preproattacin and its processing in insect cells infected with a recombinant baculovirus.from a cdna library made from fatbody of immunized hyalophora cecropia pupae, a full-length clone corresponding to basic attacin was isolated. the corresponding amino acid sequence suggests that basic attacin is synthesized as a 233-residue preproprotein. this was confirmed by cloning the cdna fragment encoding the basic attacin in the baculovirus autographa californica nuclear polyhedrosis virus downstream of the polyhedrin promoter and expressing the protein in spodoptera frugiperda (sf9) cell ...19902406140
functional analysis of a 603 nucleotide open reading frame upstream of the polyhedrin gene of autographa californica nuclear polyhedrosis virus.the sequence of the 2000 nucleotides immediately upstream of the polyhedrin gene of the autographa californica multiple nucleocapsid nuclear polyhedrosis virus has been determined. comparative analysis of the data identified a 603 nucleotide open reading frame (orf) separated from the polyhedrin gene coding sequences by 156 nucleotides and in the opposite strand of dna. northern hybridization analysis of polyadenylated rna from infected cells highlighted a 3.7 kb species produced maximally at 12 ...19902407804
insect resistant cotton plants.we have expressed truncated forms of the insect control protein genes of bacillus thuringiensis var. kurstaki hd-1(cryia(b) and hd-73 (cryia(c) in cotton plants at levels that provided effective control of agronomically important lepidopteran insect pests. total protection from insect damage of leaf tissue from these plants was observed in laboratory assays when tested with two lepidopteran insects, an insect relatively sensitive to the b.t.k. insect control protein, trichoplusia ni (cabbage loo ...19901366777
occluded and nonoccluded nuclear polyhedrosis virus grown in trichoplusia ni: comparative neutralization comparative infectivity, and in vitro growth studies.nuclear polyhedrosis virus infections of lepidopteran cells often result in the production of both occluded and nonoccluded virus. the characterization of these two different forms has been the subject of several papers. we have divided the nonoccluded virus (nov) category further into plasma membrane-budded non-occluded virus (pmb-nov), intracellular nov, and hemolymph-derived nov, and have done additional studies investigating the differences between these nonoccluded forms and the alkali-libe ...1976787558
complementation of recombinant baculoviruses by coinfection with wild-type virus facilitates production in insect larvae of antigenic proteins of hepatitis b virus and influenza virus.we describe the coinfection of insects with wild-type and recombinant baculoviruses in which the polyhedrin gene promoter is used to express hepatitis b virus envelope protein (hepatitis b virus surface antigen; hbsag) or influenza a virus neuraminidase (na). viruses were administered per os to larvae of the cabbage looper, trichoplusia ni, causing an infection that within 5 days resulted in the production of approximately 0.15 mg of hbsag per insect, representing 1.5% of the total extracted pro ...19892646635
insect cell culture: improved media and methods for initiating attached cell lines from the lepidoptera.several cell lines from the pupae of the noctuid moth species spodoptera frugiperda, heliothis zea, and trichoplusia ni were isolated on a synthetic medium containing insect hemolymph and turkey serum. these lines were progressively adapted to improved media free of insect hemolymph but containing one or more of the following sera: turkey, chicken, and fetal calf. primary culture tissue disruption was improved by substituting collagenase for trypsin. primary culture survival was improved by cont ...1975172433
characterization and serology of the matrix protein from a nuclear-polyhedrosis virus of trichoplusia ni before and after degradation by an endogenous proteinase.the intact matrix protein from a nuclear-polyhedrosis virus of the cabbage looper (trichoplusia ni), isolated after inhibition of an endogenous serine-type proteinase, was further purified by molecular-sieve chromatography. the matrix protein was associated with carbohydrate moieties, and the carbohydrate content was determined for the two major peptides isolated after proteolysis by the endogenous proteinase. the association-dissociation interactions of the intact and proteinase-hydrolysed mono ...197723107
sanguinarine, a phototoxic h2o2-producing alkaloid.sanguinarine chloride, a quaternary salt of a benzophenanthrene alkaloid, was phototoxic to catalase-deficient strains of escherichia coli but not to trichoplusia ni (cabbage looper moth larvae), an insect with high levels of catalase activity. chemical analyses confirm that sanguinarine is an efficient producer of h2o2. this differential toxicity suggests that the mode of phototoxic action involves production of h2o2 which could be detoxified in many organisms by catalase.19892696988
identification and distribution of a proctolin-like neuropeptide in the nervous system of the gypsy moth, lymantria dispar, and in other lepidoptera.although the neuropeptide proctolin has important functions in many arthropods, it is reported to be absent in lepidoptera. its possible occurrence in these insects was reinvestigated by bioassays of hplc fractions and immunocytochemistry. a proctolin-like substance was recovered from the frontal and subesophageal ganglia of lymantria dispar. this substance has the same chromatographic retention time as proctolin; enzymatic degradation indicates that it is a peptide; it is bound by proctolin ant ...19892732362
insecticidal properties of genetically engineered baculoviruses expressing an insect juvenile hormone esterase gene.exploring the possibility of enhancing the properties of baculoviruses as biological control agents of insect pests, we tested the effect of expressing an insect gene (jhe) encoding juvenile hormone esterase. juvenile hormone esterase inactivates juvenile hormone, which regulates the outcome of an insect molt. a cdna encoding the juvenile hormone esterase of heliothis virescens was inserted into the genome of autographa californica nuclear polyhedrosis virus such that the gene was expressed unde ...19921622228
comparisons of host cell dna insertions and altered transcription at the site of insertions in few polyhedra bacilovirus mutants.few polyhedgra (fp) mutants of autographa californica nuclear polyhedrosis virus (acnpv) and the closely related strain galleria mellonella (gm)npv have been reported which contain trichoplusia ni host cell dna sequences inserted into the viral genome between map units 35.0 and 37.7. new fp mutants are described with alterations of the hindiii-i restriction enzyme fragment (33.8 to 37.7 map units) of acnpv, either deletions of viral dna sequences or insertions of spodoptera frugiperda host cell ...19882829419
effects of serial passage of autographa californica nuclear polyhedrosis virus in cell culture.a study of the major genomic alterations occurring during serial passage of autographa californica nuclear polyhedrosis virus (acnpv) in a trichoplusia ni cell line was conducted. progeny viruses from 24 independent passages were randomly selected and analyzed with restriction endonucleases. specific deletion mutations were generated repeatedly in the psti-g (7.6 to 13.1%) and the psti-i (14.4-17.9%) regions; these mutations became predominant in the serially passaged stocks in which they arose. ...19872887077
transposon-mediated mutagenesis of a baculovirus.spontaneous mutants of insect nuclear polyhedrosis viruses of autographa californica (acmnpv) and galleria mellonella (gmmnpv) were analyzed. these mutants produce few polyhedra in infected cells and have insertions of host cell dna. all the insertions mapped to two adjacent sites in the genome. the junctions between two host insertions and the viral dna were sequenced. one of the insertions contained a perfect 7-bp inverted terminal repeat, and had caused a direct duplication of 4 bp of viral d ...19852992159
co-expression of the hepatitis b surface and core antigens using baculovirus multiple expression vectors.the hepatitis b (hb) virus dna sequences coding for the pre-core (prec) or c antigens (hbpcag, hbcag) have been inserted into the baculovirus plasmid transfer vector, pacym1, such that the hb viral sequences are under the control of the polyhedrin promoter of autographa californica nuclear polyhedrosis virus (acnpv). spodoptera frugiperda cells infected with either of the derived recombinant plasmids in the presence of infectious acnpv dna yielded recombinant, polyhedrin-negative viruses that ex ...19883053987
expression of post-translational processing of preprocecropin a using a baculovirus vector.a cdna fragment encoding preprocecropin a was inserted into the baculovirus autographa californica nuclear polyhedrosis virus downstream of the polyhedrin promoter. the gene was expressed in recombinant-infected last instar larvae of trichoplusia ni and in diapausing pupae of hyalophora cecropia. the identity of the recombinant product was established by electrophoresis with detection of antibacterial activity and mass spectrometry. the prepropeptide had been correctly processed including remova ...19911712729
protective effect of methylcellulose and other polymers on insect cells subjected to laminar shear stress.the relative sensitivity of two insect cell lines to laminar shear stress was determined, and the protective effect of polymers added to the growth media of two insect cell lines, trichoplusia ni (tn-368) and spodoptera frugiperda (sf-9), was evaluated. tn-368 and sf-9 cells were found to be equally sensitive to laminar shear stress. methylcellulose [0.5% (w/v) dow e4m methocel] and dextran [4.5% (w/v)] increased the resistance of suspended cells to lysis due to laminar shear stress by factors o ...19901370016
the development of multiple expression vectors for high level synthesis of eukaryotic proteins: expression of lcmv-n and acnpv polyhedrin protein by a recombinant baculovirus.a copy of the polyhedrin gene promoter of autographa californica nuclear polyhedrosis virus (acnpv), in association with the coding region of lymphocytic choriomeningitis virus n protein (lcmv-n) and the relevant polyhedrin transcription termination signals, has been cloned into the unique ecorv site of a plasmid representing an ecori derived fragment of the acnpv genome. the cloning site is upstream (but in the opposite orientation) of the natural acnpv polyhedrin gene. the derived pacvc2 trans ...19873334094
functional domains of bacillus thuringiensis insecticidal crystal proteins. refinement of heliothis virescens and trichoplusia ni specificity domains on cryia(c).insecticidal crystal proteins (delta-endotoxins), cryia(a) and cryia(c), from bacillus thuringiensis are 82% homologous. despite this homology, cryia(c) was determined to have 10-fold more insecticidal activity toward heliothis virescens and trichoplusia ni than cryia(a). reciprocal recombinations between these two genes were performed by the homolog-scanning technique. the resultant mutants had different segments of their primary sequences exchanged. bioassays with toxin proteins from these mut ...19911917934
synthesis and toxicity of full-length and truncated bacterial cryivd mosquitocidal proteins expressed in lepidopteran cells using a baculovirus vector.full-length (72k) and truncated (61k) cryivd mosquitocidal proteins of bacillus thuringiensis (bt) were expressed in spodoptera frugiperda cells and larvae of trichoplusia ni using a baculovirus vector to investigate the role of cryivd peptides in toxicity as well as to evaluate further the baculovirus/lepidopteran system for expressing bt proteins. the cryivd genes were inserted into the autographa californica multinucleocapsid nuclear polyhedrosis virus (acmnpv) under control of the polyhedrin ...19921730944
fatty acid composition of whole bodies, specific tissues and cell lines of two lepidopteran insects.we detail the fatty acid compositions of last larval instars of two lepidopterans, spodoptera frugiperda and trichoplusia ni, two tissues from t. ni, a cell line derived from each species and the respective larval and cell culture media. larval whole-body and specific tissue fatty acid profiles exhibited the major features commonly found in previous lepidopteran analyses, whereas the cell-line fatty acid compositions were substantially different from the compositions of both their growth media a ...19863780187
localization of juvenile hormone esterase during development in normal and in recombinant baculovirus-infected larvae of the moth trichoplusia ni.the pathogenesis and cellular localization of juvenile hormone esterase (jhe) was examined in larvae of the moth trichoplusia ni, infected with a recombinant baculovirus (autographa californica nuclear polyhedrosis virus: acnpv) engineered to produce high levels of jhe (jhe virus). the course of jhe localization in the recombinant virus infected larvae was compared with that of both wild type acnpv infected, and uninfected larvae, using immunogold electron microscopy. in the jhe virus infected i ...19921589874
serological studies on virions and polyhedron protein of a nuclear polyhedrosis virus of the cabbage looper, trichoplusia ni. 1977404757
parasite regulation of host insect metamorphosis: a new form of regulation in pseudoparasitized larvae of trichoplusia ni.when eggs of trichoplusia ni (lepidoptera) are stung by a parasitic wasp, chelonus sp., the developing host larvae precociously initiate metamorphosis ten days later. precocious initiation of metamorphosis occurs even in 'pseudoparasitized' stung hosts which contain no living parasites at the time of symptoms of host regulation by the parasite. in feeding, penultimate instar, pseudoparasitized hosts, the corpora allata activity, hemolymph juvenile hormone esterase activity, in vivo rates of juve ...19853837029
replication of autographa californica baculovirus (acmnpv) in a coleopteran cell line.a coleopteran cell line (age) derived from the cotton boll weevil anthonomus grandis supported replication of autographa californica multiple nuclear polyhedrosis virus (acmnpv). the titer of extracellular virus (ecv) and the number of occlusion bodies (ob) produced in age cells were approximately equal to those produced by a trichoplusia ni cell line (tn-cl1), and the ob produced by both cell lines were equally infectious for t. ni larvae. the identity of the age cell line was established by ch ...19921522049
in vitro host range of five baculoviruses in lepidopteran cell lines.the in vitro host range of five nuclear polyhedrosis viruses (npv) was assessed in five lepidopteran cell lines from three genera. multiple-enveloped baculoviruses of autographa californica (acmnpv), trichoplusia ni (tnmnpv), and galleria mellonella (gmmnpv) replicated in cells of t. ni, spodoptera frugiperda, and heliothis virescens to a titer of approximately 10(7) tcid50/ml. the multiple-enveloped baculovirus of s. frugiperda (sfmnpv) replicated only in s. frugiperda cells. the single-envelop ...19853988484
hemagglutinin activity in the hemolymph of anticarsia gemmatalis larvae infected with the fungus nomuraea rileyi.hemolymph samples from larvae of 3 lepidopteran species (anticarsia gemmatalis, trichoplusia ni and spodoptera frugiperda) were tested for hemagglutination activity. samples from a. gemmatalis larvae which had been injected 12-24 hrs previously with hyphal bodies of the entomopathogenic fungus nomuraea rileyi showed agglutination activity against human 0, rabbit and sheep erythrocytes. little or no activity was detected in samples from the other 2 larval species. low titers (approximately 1:2) w ...19853996705
baculovirus expression of alkaline phosphatase as a reporter gene for evaluation of production, glycosylation and secretion.we have devised a simple and efficient baculovirus expression vector system to evaluate insect tissue culture cells for their capacity to express, glycosylate and secrete foreign proteins. a truncated placental alkaline phosphatase (seap) gene was inserted into the autographa californica nuclear polyhedrosis virus (acmnpv) genome under the transcriptional control of the polyhedrin gene promoter. production levels, glycosylation, and secretion of the recombinant protein were examined in trichoplu ...19921368794
differentiation of trichoplusia ni mev and autographa californica mev by macrophage migration inhibition tests. 19734578231
differential susceptibility of parasitized and nonparasitized larvae of trichoplusia ni to a nuclear polyhedrosis virus. 19744606436
some biophysical properties of virus present in tissue cultures infected with the nuclear polyhedrosis virus of trichoplusia ni. 19744617765
characterization of deoxyribonucleic acid isolated from the granulosis viruses of the cabbage looper, trichoplusia ni and the fall armyworm, spodoptera frugiperda. 19724636117
characterization of nuclear polyhedrosis virus dnas.the nuclear polyhedrosis virus dnas characterized and compared in this study consist of the singly-enveloped nucleocapsids (snpv) of trichoplusia ni and the bundles of nucleocapsids common to a single envelope (mnpv) from spodoptera frugiperda and rachiplusia ou. the snpv and mnpv dnas are very similar in hydrodynamic properties and molecular weights. in addition, the npv dnas are similar in size to those extracted from the granulosis viruses that infect t. ni and s. frugiperda. as isolated from ...19734761726
strain selection during serial passage of trichoplusia in nuclear polyhedrosis virus.two strains of a nuclear polyhedrosis virus (npv) of trichoplusia ni were isolated on the basis of plaque morphology. they are designated as mp (having greater than 30 polyhedra per nucleus) and fp (having fewer than 10 polyhedra per nucleus). serial, undiluted passage of plaque, purified mp nonoccluded. virus (nov) in tissue culture led to the production of the fp phenotype detectable at passage 9. with continued serial, undiluted passage, fp became the predominant strain. comparative growth cu ...1976775129
recovery of the nuclear polyhedrosis virus of the cabbage looper, trichoplusia ni, by coprecipitation with lactose. 19705449744
characterization of the da26 gene in a hypervariable region of the autographa californica nuclear polyhedrosis virus genome.a region of the baculovirus autographa californica nuclear polyhedrosis virus genome that is frequently found to be altered after serial passage of the virus in cell culture was characterized. sequence analysis of this region of the genome in wild-type and mutant viruses revealed that some of the mutations affected a 675 bp open reading frame, designated da26. the da26 gene was disrupted both by deletion and by insertion of sequences that resembled transposable elements. northern blot analysis o ...19902189022
observations on an artifact present in trichoplusia ni granulosis-virus preparations. 19685725010
sex pheromones of noctuid moths. v. circadian rhythm of pheromone-responsiveness in males of autographa californica, heliothis virescens, spodoptera exigua, and trichoplusia ni (lepidoptera: noctuidae). 19655834924
exposure of white mice and guinea pigs to the nuclear-polyhedrosis virus of the cabbage looper, trichoplusia ni. 19665905543
spicaria rileyi (farlow) charles and aspergillus flavus link, entomogenous fungi of the cabbage looper, trichoplusia ni (hübner), in indiana and wisconsin. 19665948338
the antiviral spectrum of (e)-5-(2-bromovinyl)-2'-deoxyuridine.the antiviral activity spectrum of (e)-5-(2-bromovinyl)-2'-deoxyuridine (bvdu) is not restricted to herpes simplex virus type 1 (hsv-1) and varicella-zoster virus (vzv) but also encompasses several other herpesviruses such as suid herpesvirus type 1 (shv-1), bovid herpesvirus type 1 (bhv-1), simian varicella virus (svv), herpesvirus saimiri, herpesvirus platyrrhinae, and the baculovirus trichoplusia ni multiple nuclear polyhedrosis virus. other herpesviruses such as herpes simplex virus type 2, ...19846092320
efficient, low-cost protein factories: expression of human adenosine deaminase in baculovirus-infected insect larvae.human adenosine deaminase (ec 3.5.4.4), a key purine salvage enzyme essential for immune competence, has been overproduced in spodoptera frugiperda cells and in trichoplusia ni (cabbage looper) larvae infected with recombinant baculovirus. the coding sequence of human adenosine deaminase was recombined into a baculovirus immediately downstream from the strong polyhedrin gene promoter. approximately 60 hr after infection of insect cells with the recombinant virus, maximal levels of intracellular ...19902181448
heterocyclic derivatives of 3-substituted-1,1,1-trifluoro-2-propanones as inhibitors of esterolytic enzymes.a series of (alkylthio)trifluoropropanones containing a heterocyclic moiety was synthesized. the compounds were tested for in vitro inhibition of four hydrolytic enzymes including insect juvenile hormone esterase (jhe), eel acetylcholinesterase (ache), yeast lipase (lp), and bovine alpha-chymotrypsin. the i50 values ranged from 10(-3) to 10(-7) m. 3-(2-pyridylthio)-1,1,1-trifluoro-2-propanone was found to be the most potent inhibitor as compared to the other tested heterocyclic analogues with an ...19902133080
screening of insect cell lines for the production of recombinant proteins and infectious virus in the baculovirus expression system.eight cell lines derived from the insects spodoptera frugiperda, trichoplusia ni, mamestra brassicae, and estigmene acrea were evaluated for recombinant beta-galactosidase and infectious virus production following infection with the baculovirus autographa californica multiple nuclear polyhedrosis virus (acmnpv). production was assessed on a specific (per cell and per microgram of uninfected cellular protein) and on a volumetric (per milliliter) basis. cell density was found to be an important fa ...19921369220
baculovirus replication: phosphorylation of polypeptides synthesized in trichoplusia ni nuclear polyhedrosis virus-infected cells.a number of polypeptides synthesized specifically in trichoplusia ni multiple nucleocapsid nuclear polyhedrosis virus (t. ni mnpv)-infected spodoptera frugiperda cells are phosphorylated both early and late in infection. certain non-structural proteins and the major basic internal protein are the main phosphoproteins detected in infected cells. the polyhedron protein was not phosphorylated. many cell proteins continue to be phosphorylated throughout infection. pulse-chase experiments have shown ...19846379101
trichoplusia ni granulosis virus granulin: a phenol-soluble, phosphorylated protein.trichoplusia ni granulosis virus granulin consists of one major polypeptide component with an estimated molecular weight of 28,000. the protein is phenol soluble, phosphorylated, and acidic. a protease activated by alkaline conditions is also associated with solubilized granulin preparations. if not properly inactivated, the protease will introduce extensive artifact into the protein giving rise to ambiguous and incorrect results as analyzed by sds-polyacrylamide gel electrophoresis and peptide ...19751185849
nuclear polyhedrosis virus detection: relative capabilities of clones developed from trichoplusia ni ovarian cell line tn-368 to serve as indicator cells in a plaque assay.cloned cell lines from the established trichoplusia ni line tn-368 appear to differ from one another in their relative capabilities to serve as plaque assay indicator cell lines for autographa californica nuclear polyhedrosis virus. although there seems to be little correlation between their relative generation times and their efficiency in supporting plaque formation as indicator cell lines, there does seem to be a relationship within a given line between its capability to serve as an indicator ...19751104895
bacillus thuringiensis var israelensis crystal delta-endotoxin: effects on insect and mammalian cells in vitro and in vivo.bacillus thuringiensis var israelensis parasporal crystal delta-endotoxin was purified by ultracentrifugation on a discontinuous sucrose gradient. native delta-endotoxin crystals showed no detectable toxicity in the vitro and in vivo systems that are described. by contrast alkali-solubilized crystal delta-endotoxin caused rapid cytological and cytopathological changes in aedes albopictus, choristoneura fumiferana 63 cf1, spodoptera frugiperda and trichoplusia ni cell lines as observed by phase-c ...19836874728
release of pi-anchoring enzymes and other effects of phosphatidylinositol-specific phospholipase c from bacillus thuringiensis on tn-368 cells from a moth ovary.release of pi-anchoring enzymes and other effects of phosphatidylinositol-specific phospholipase c from bacillus thuringiensis on tn-368 cells from a moth ovary. toxicon 27, 637-645, 1989.--the effect of phosphatidylinositol-specific phospholipase c(piplc) from bacillus thuringiensis was investigated on tn-368 cells, derived from the ovary of a moth, trichoplusia ni. quantitative analysis of lipids showed that phosphatidylinositol (pi) was contained as one of the major phospholipids in tn-368 ce ...19892749761
simple method for the isolation of the antilepidopteran toxin from bacillus thuringiensis subsp. kurstaki.a protein with a molecular mass of 66 kda was isolated by a simple, rapid, and inexpensive method, using 3-n-morpholinopropanesulfonic acid, potassium thiocyanate, and dithiothreitol, from a mixture of spores, parasporal crystals, and cell debris of bacillus thuringiensis subsp. kurstaki. the protein was active against the third instar larvae of trichoplusia ni, was soluble in 19 mm na2co3, and was characterized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and confirmed as the in ...19902360990
relative activity of bacillus thuringiensis var. kurstaki and b. thuringiensis var. israelensis against larvae of aedes aegypti, culex quinquefasciatus, trichoplusia ni, heliothis zea, and heliothis virescens. 19817320315
newly identified, basic hemolymph proteins from noctuid species.a number of basic metamorphosis-associated proteins were identified from several noctuid species. all of these proteins have molecular weights in the range of 73,000 to 74,000. two of the proteins in trichoplusia ni and heliothis virescens were found to be suppressible by a juvenile hormone analog.19873606619
alteration of a lepidopteran peritrophic membrane by baculoviruses and enhancement of viral infectivity.the peritrophic membrane (pm), which lines the midgut of many insect species, has several functions. in particular, it may serve as a mechanical barrier to invading microorganisms. the protein composition of the pm from healthy and baculovirus-treated trichoplusia ni (cabbage looper) larvae was analyzed by polyacrylamide gel electrophoresis. a specific interaction took place between baculoviruses and the pm of susceptible t. ni larvae. a 68-kda glycoprotein of the pm disappeared within 15 min po ...19883055665
immunochemical characterization of juvenile hormone esterase from different species of lepidoptera.cross-immunoreactivity of juvenile hormone esterase (jhe) from different species was tested using anti-jhe (trichoplusia ni) (noctuidae) polyclonal antibody. partial cross-reactivity was observed between jhe from hyphantria cunea, isia isabella (arctuidae) and spodoptera exigua (noctuidae) in immunoblot analysis. soluble antigen-antibody complex formation was observed between anti-jhe (t. ni) and antigen(s) from heliothis virescens (noctuidae) during immunotitration of antigen(s). using an elisa ...19873579958
an insect model for assessing oxidative stress related to arsenic toxicity.the potential usefulness of an insect model to evaluate oxidative stress induced by environmental pollutants was examined with trivalent arsenic (as3+, naaso2) and pentavalent arsenic (as5+, na2haso4) in adult female house flies, musca domestica, and fourth-instar cabbage loopers, trichoplusia ni. m. domestica was highly susceptible to both forms of arsenic following 48 h exposure in the drinking water with lc50s of 0.008 and 0.011% w/v for as3+ and as5+, respectively. t. ni larvae were suscepti ...19957606044
another version of the human insulin receptor kinase domain: expression, purification, and characterization.we have overexpressed another insulin receptor kinase molecule, which consists of residues 941-1343 inclusive of the human insulin receptor, by using the baculo-virus expression vector pvl941. unlike the two previous preparations of insulin receptor kinase in this expression system, this molecule contains the complete unmodified sequence of the cytoplasmic domain of the human insulin receptor beta subunit. our construct allows high-level expression of the recombinant protein in cultured sf9 cell ...19892554291
horseradish peroxidase phe172-->tyr mutant. sequential formation of compound i with a porphyrin radical cation and a protein radical.a gene coding for the f172y mutant of horseradish peroxidase isozyme c (hrp) has been constructed and expressed in both spodoptera frugiperda (sf-9) and trichoplusia ni egg cell homogenate (highfive) cells. homology modeling with respect to three peroxidases for which crystal structures are available places phe172 on the proximal side of the heme in the vicinity of porphyrin pyrrole ring c. the ph optimum and spectroscopic properties of the f172y mutant are essentially identical to those of wild ...19957629167
multicellular-vesicle-promoting polypeptide from trichoplusia ni: tissue distribution and n-terminal sequence.an n-terminal amino acid sequence of a 16.9 kda hemolymph polypeptide, "vesicle promoting factor" (vpf) from trichoplusia ni, revealed a high sequence homology (70%) with manduca sexta apolipophorin-iii. a polyclonal antibody developed against vpf, however, was not immunoreactive with either purified m. sexta or t. ni apolipophorin-iii. immunoblots of tissue homogenates of t. ni indicated that vpf was present in imaginal wing discs, central nervous system (cns), silk glands, midgut and hemocytes ...19957655058
differential requirements for baculovirus late expression factor genes in two cell lines.a plasmid library of 18 late expression factor (lef) genes (lef library) from the baculovirus autographa californica nuclear polyhedrosis virus (acmnpv) supports transient expression from a late viral promoter in the sf-21 cell line, derived from spodoptera frugiperda. we found, however, that this lef library was unable to support expression from the same promoter in the tn-368 cell line, derived from trichoplusia ni, which is also permissive for acmnpv replication. to identify the additional fa ...19957666527
antigenic differences between european and american isolates of porcine reproductive and respiratory syndrome virus (prrsv) are encoded by the carboxyterminal portion of viral open reading frame 3.antigenic differences between european and american isolates of porcine reproductive and respiratory syndrome virus (prrsv) were revealed by serologic analysis of a recombinant protein derived from prrsv open reading frame 3 (orf 3). the hydrophilic carboxyterminal 199 amino acids encoded by the orf 3 of a european (lelystad) isolate of prrsv were expressed as a recombinant fusion protein (bp03-p) in a baculovirus gene expression system. sera from gnotobiotic swine exposed to prototypic referenc ...19957667907
high level expression and phosphorylation of hepatitis b virus polymerase in insect cells with recombinant baculoviruses.the hepatitis b virus polymerase open reading frame, as well as various subdomains of polymerase, was expressed in insect cells using the recombinant baculovirus expression system. full-length polymerase was expressed at very low levels in a spodoptera frugiperda cell line, the amino-terminal domain of polymerase was expressed at high levels, and other constructs were expressed at intermediate levels. infections of a trichoplusia ni cell line with the same recombinant baculoviruses resulted in h ...19937683161
the pathway of infection of autographa californica nuclear polyhedrosis virus in an insect host.an immunohistochemical study was conducted to detect the temporal infection sequence of autographa californica m nuclear polyhedrosis virus in trichoplusia ni larvae. staining patterns indicated that the initial infection occurred in the midgut, simultaneously in columnar epithelial and regenerative cells, but that subsequently this tissue recovered. a major envelope glycoprotein stained in a polar fashion when it was expressed in columnar epithelial cells, but not when expressed in other cells ...19892648574
the inactivation of the nuclear polyhedrosis virus of trichoplusia ni by gamma and ultraviolet radiation. 19685681529
rate of increase of autographa californica nuclear polyhedrosis virus in trichoplusia ni larvae determined by dna:dna hybridization.the rate of increase and doubling time of the hob clone of autographa californica nuclear polyhedrosis virus (acmnpv-hob) in neonate trichoplusia ni larvae was determined by measuring the increase in viral dna through time following inoculation with average doses of 50 or 17,400 occlusion bodies per larva. changes in total dna and viral dna through time were followed by fluorescence spectroscopy and quantitative slot-blot dna:dna hybridization, respectively. total dna content (i.e., larval dna a ...19902405064
baculovirus replication: inhibition of trichoplusia ni multiple nuclear polyhedrosis virus by [e]-5-(2-bromovinyl)-2'-deoxyuridine.[e]-5-(2-bromovinyl)-2'-deoxyuridine (bvdu) inhibits the replication of the baculovirus trichoplusia ni multiple nucleocapsid nuclear polyhedrosis virus in spodoptera frugiperda cells. virus-specific dna synthesis and late protein synthesis are suppressed by the drug. bvdu is phosphorylated by deoxythymidine (deoxycytidine) kinase present in both uninfected and virus-infected cells, and in its 5'-triphosphate form it inhibits dna polymerase activity in virus-infected cells. the effect of the bvd ...19836343553
rate of increase and critical amount of nuclear polyhedrosis virus in lepidopterous larvae estimated from survival time assay data with a birth-death model.a birth-death model developed for pathogens of vertebrates was used to estimate the in vivo rate of increase (alpha) and the doubling time (td) from survival time assay data. host-pathogen combinations used in this study were two autographa californica nuclear polyhedrosis virus isolates in trichoplusia ni and heliothis zea npv in h. zea. the alpha's, estimated as he negative reciprocal of the slope of the linearly decreasing section of the plot of median survival times against the logarithm of ...19883291823
optimization of growth methods and recombinant protein production in bti-tn-5b1-4 insect cells using the baculovirus expression system.a novel insect cell line from trichoplusia ni, bti-tn 5b1-4 (tn 5), was compared to spodoptera frugiperda, sf 9, cells for production of two recombinant secreted proteins: truncated epstein-barr viral attachment protein (ebv gp105) and truncated, soluble tissue factor (stf). under optimum conditions for both cell lines, tn 5 cells produced 28-fold more secreted stf than sf 9 cells, respectively, on a per cell basis. the total production of gp105 was similar for the two cell lines. however, tn5 c ...19937764044
production of human alkaline phosphatase, a secreted, glycosylated protein, from a baculovirus expression system and the attachment-dependent cell line trichoplusia ni bti-tn 5b1-4 using a split-flow, air-lift bioreactor.a split-flow, air-lift bioreactor for the cultivation of insect cells to produce recombinant protein is described. it can be used advantageously with attached cell systems. this bioreactor incorporates two sections: a rise and a downcomer. trichoplusia ni bti-tn 5b1-4 cells are grown on a support material of glass beads or microcarriers placed in the downcomer. this cell line is more productive than other commonly used insect cell lines, but it has the disadvantage of being difficult to use at l ...19937764358
gene organization and transcription of ted, a lepidopteran retrotransposon integrated within the baculovirus genome.a single copy of the retrotransposon ted, from the moth trichoplusia ni (a lepidopteran noctuid), was identified within the dna genome of the baculovirus autographa californica nuclear polyhedrosis virus. determination of the complete nucleotide sequence (7,510 base pairs) of the integrated copy indicated that ted belongs to the family of retrotransposons that includes drosophila melanogaster elements 17.6 and gypsy and thus represents the first nondipteran member of this invertebrate group to b ...19901692964
developmental resistance in fourth instar trichoplusia ni orally inoculated with autographa californica m nuclear polyhedrosis virus.larvae of lepidopteran insects commonly become increasingly resistant to baculovirus infections as they age. the mechanism responsible for this development resistance is not known, but the phenomenon does not occur if the viral inoculum is administered intrahemocoelically instead of orally, which is the natural route of infection. this observation indicates that the factors mediating developmental resistance are operative during infection of the primary target tissue, the larval midgut, and not ...19957778273
the nucleotide sequence of the pieris brassicae granulosis virus granulin gene.two overlapping restriction fragments containing the pieris brassicae granulosis virus (gv) granulin gene were cloned into plasmids. the regions containing the coding region and the 5' and 3' flanking regions were subcloned into m13 and sequenced. the nucleotide sequence data were compared to those for the granulin gene from the trichoplusia ni gv and the polyhedrin gene from the autographa californica nuclear polyhedrosis virus (acmnpv). the amino acid sequences derived from these dna sequences ...19853891914
lack of functional significance of cys227 and cys234 in terminal deoxynucleotidyltransferase.identification of the three functional regions (catalytic, nucleotide substrate-binding, dna substrate-binding) of the monofunctional template independent dna polymerase terminal deoxynucleotidyltransferase has not been completely established. the potential participation of 2 amino acid residues, cys227 and cys234, has been controversial, and conflicting data have been published. to investigate the role of cys227, the human terminal transferase cdna was modified by site-directed mutagenesis to i ...19921544903
physical and chemical properties of trichoplusia ni granulosis virus granulin.the protein solubilized from the proteinic crystalline structure surrounding the granulosis virus of trichoplusia ni by use of a carbonate buffer (ph 10.7) gives a major component, as analyzed by ultracentrifugation, with a molecular weight of 180,000. this protein has heterogeneous subunit structure as demonstrated by estimates of molecular weights by use of gel electrophoresis, amino-, and carboxy-terminal analyses, and peptide mapping of enzyme digests of the protein. the amino acid compositi ...19734765397
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