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immunochemical characterization of lymantri dispar npv hemagglutinin: protein-carbohydrate interaction.the agglutination of chicken erythrocytes by lymantria dispar nuclear polyhedrosis virus polyhedrin has been shown to provide specific virus identification. selected mono- and oligosaccharides, present in blood group substances, were assayed by the land-steiner hapten inhibition technique for specific inhibition of polyhedrin hemagglutination. n-acetylgalactosamine and n-acetylglucosamine inhibit to the greatest extent; galactosamine, glucosamine and fucose to a lesser extent. the hapten inhibit ...197822517
[detection of carriers of latent nuclear polyhedrosis virus in natural populations of the gypsy moth lymantria dispar l. (lepidoptera, family lymantriidae)]. 1977140992
gypsy moth cell lines divergent in viral susceptibility. i. culture and identification.a series of cell lines unique in insect virus susceptibility pattern have been isolated from the ovaries of the gypsy moth (lymantria dispar: lepidoptera: lymantriidae) on a synthetic medium with mammalian and avian serum supplementation. growth curves showed the poorest growth occurring on peptone-based media with somewhat better growth on amino-acid-based media. the best growth was obtained with combined media. serological study distinguished the present cell lines from one another and from ce ...197879542
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
identification and characterization of the ie-1 gene of orgyia pseudotsugata multicapsid nuclear polyhedrosis virus.the ie-1 gene of orgyia pseudotsugata multicapsid nuclear polyhedrosis virus (opmnpv) was mapped between 95.7 and 97.1 map units on the viral genome. sequence analysis of the opmnpv ie-1 gene (opie-1) identified an open reading frame that coded for a predicted protein of 560 amino acids with a molecular weight of 64,775. transcriptional analysis of opmnpv-infected lymantria dispar (ld652y) cells identified two rnas homologous to the opie-1 open reading frame that were 1.7 and 1.9 kb in size. the ...19911989381
a baculovirus polyhedron envelope protein: immunogold localization in infected cells and mature polyhedra.a polyclonal antiserum against a trpe fusion protein containing the complete open reading frame of the polyhedron envelope (pe) protein from the nuclear polyhedrosis virus of orgyia pseudotsugata (opmnpv) was used for immunogold staining and electron microscopic examination of polyhedra, isolated polyhedron envelopes, and infected insect cells at selected times postinfection. the antiserum specifically stained the peripheral envelope of mature polyhedra and also stained the envelope structure wh ...19902403704
immunoelectron microscopic examination of orgyia pseudotsugata multicapsid nuclear polyhedrosis virus-infected lymantria dispar cells: time course and localization of major polyhedron-associated proteins.immunoelectron microscopy was employed to examine the temporal expression and localization of two proteins involved in baculovirus polyhedron assembly (polyhedrin and p10) of orgyia pseudotsugata multicapsid nuclear polyhedrosis virus (opmnpv) in infected lymantria dispar cells. in addition, the association of p10 with the polyhedron envelope (pe) protein was studied. the major capsid protein (p39) was also examined to investigate the association of virion structural proteins with polyhedron for ...19911993872
baculovirus gp64 gene expression: analysis of sequences modulating early transcription and transactivation by ie1.expression of the baculovirus major envelope glycoprotein gene (gp64) is regulated by transcription from both early and late promoters. to characterize the early promoter and identify sequences involved in the regulation of gp64 early transcription, promoter-reporter gene fusions were generated from the orygia pseudotsugata nuclear polyhedrosis virus gp64 promoter and were analyzed by transient expression in uninfected insect cells. for these analyses, 5' deletion mutations were constructed in t ...19911920618
field evaluation of a dna hybridization assay for nuclear polyhedrosis virus in gypsy moth (lepidoptera: lymantriidae) larvae.dna hybridization assays were used to detect the presence of viral dna in gypsy moth (lymantria dispar l.) larvae collected weekly from high density populations or reared from field-collected egg masses. dna was extracted from larvae, bound to nitrocellulose filters, and hybridized with digoxigenin-labeled l. dispar npv (ldnpv) dna probes. the virus incidence determined from dna hybridization assays was compared with that determined with conventional microscopic examination of larvae for polyhed ...19911842802
tandemly repeated sequence at the 3' end of the ie-2 gene of the baculovirus orgyia pseudotsugata multicapsid nuclear polyhedrosis virus is an enhancer element.an enhancer element was identified in the virus orgyia pseudotsugata multicapsid nuclear polyhedrosis virus (opmnpv) that is located adjacent to the 3' end of the ie-2 gene and 5' to an open reading frame that codes for a predicted protein that has 37% homology to the acmnpv pe-38 gene. the opmnpv enhancer (ope) consists of a 66-bp element that is tandemly repeated partially or completely 12 times. the ope sequences were shown to increase gene expression from the autographa californica mnpv dela ...19921736547
changes in macromolecular synthesis of gypsy moth cell line iplb-ld652y induced by autographa californica nuclear polyhedrosis virus infection.the aberrant replication of the autographa californica multiple-enveloped nuclear polyhedrosis virus (acmnpv) in the lymantria dispar cell line iplb-ld652y was used as a model system for the investigation of factors regulating baculovirus host specificity. a previous study of this system indicates that viral gene expression in infected cells is extremely attenuated and subsequently all cellular and viral protein synthesis is inhibited. in the present study, infection of iplb-ld652y cells with ac ...19911709681
isolation of high-molecular-weight dna from insects.a simple and rapid method for the isolation of high-molecular-weight dna from insects is described. the method does not require cscl ultracentrifugation or extensive dialysis. high-molecular-weight dna was obtained within 24 h. since the entire insect was used for dna isolation, an initial nuclei-enriched fraction was required. genomic dna was extracted from lysed nuclei by organic phase separation (liquid/liquid extraction). this method has been successfully applied to the isolation and purific ...19901693047
genetic engineering of a lymantria dispar nuclear polyhedrosis virus for expression of foreign genes.a bacterial lacz gene was inserted into an isolate of the lymantria dispar nuclear polyhedrosis virus (ldmnpv). the transfer vector was constructed by site-directed mutagenesis of the translation start site of the ldmnpv polyhedrin gene, within the bglii e fragment of the viral genome. a multiple cloning sequence was inserted at this start site and used for the insertion of the lacz gene into the transfer plasmid. liposome transfection was used to cotransfect l. dispar tissue culture cells with ...19921607870
nucleotide sequence of the p39-capsid gene region of the lymantria dispar nuclear polyhedrosis virus.a 1.85 kb region, containing an open reading frame (orf) homologous to the baculovirus p39-capsid gene, was sequenced from the lymantria dispar multicapsid nuclear polyhedrosis virus (ldmnpv) genome. analysis of the p39-capsid gene demonstrated that it was 39% and 47% identical in amino acid sequence with the homologous genes in the autographa californica and orgyia pseudotsugata mnpvs, respectively. two late promoter elements located upstream of the p39 gene in the ldmnpv genome are conserved w ...19921607869
nucleotide sequence of the polyhedron envelope protein gene region of the lymantria dispar nuclear polyhedrosis virus.a 6.4 kb region from the lymantria dispar multicapsid nuclear polyhedrosis virus (ldmnpv) genome was sequenced and found to contain open reading frames (orfs) homologous to the polyhedron envelope (pe) protein coding sequence, and the c-terminal half of orf 1, which is a gene located upstream of the pe protein gene in other baculoviruses. the proteins predicted from the ldmnpv genes encoding the pe protein, and orf 1 demonstrated 27 and 34% amino acid sequence identity, respectively, with the co ...19921607868
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
equilibrium and kinetic analysis of autographa californica nuclear polyhedrosis virus attachment to different insect cell lines.the kinetic and equilibrium attachment of autographa californica nuclear polyhedrosis virus (acmnpv) to seven insect cell lines was evaluated. kinetic experiments revealed differences of up to 10-fold in the infection rates among cell lines. equilibrium binding also varied between cell lines and was saturable. the tn 5b1-4 and tn f cell lines had the highest virus binding affinities and infection rates and exhibited diffusion-limited attachment. the rate of infection appears to be limited by the ...19921469356
characterization of the nucleotide sequence of the lymantria dispar nuclear polyhedrosis virus dna polymerase gene region.the dna polymerase gene of the lymantria dispar multinucleocapsid nuclear polyhedrosis virus (ldmnpv) was cloned and sequenced. the predicted dna polymerase protein (1113 amino acids, 115.9k) was found to have an amino acid identity of 48% with the corresponding gene of the autographa californica mnpv (acmnpv). it contains five domains associated with substrate binding, primase interaction, and pyrophosphate hydrolysis and three domains associated with 3'-->5' exonuclease activity common to othe ...19921469355
nucleotide sequence and transcriptional analysis of the polyhedrin gene of spodoptera exigua nuclear polyhedrosis virus.the nucleotide sequence of a 1.1 kbp fragment of the multiple nucleocapsid nuclear polyhedrosis virus (mnpv) of spodoptera exigua (se) containing the polyhedrin gene was determined. an open reading frame (orf) of 738 nucleotides (nt) was detected. this orf encoded a protein of 246 amino acids with a predicted m(r) of 29k. the nucleotide and amino acid sequences were compared with the sequences of eight other npv polyhedrins. the semnpv polyhedrin protein was most closely related to s. frugiperda ...19921431809
biochemical characterization and time-course analysis of lymantria dispar nuclear polyhedrosis virus with monoclonal antibodies.hybridoma cell lines secreting monoclonal antibodies (mabs) specific to a 31,000 molecular weight viral protein or a 31,000 molecular weight polyhedrin protein of lymantria dispar nuclear polyhedrosis virus (ldnpv) were developed. the two polypeptides were shown to be different by comparing their amino acid compositions. immuno-electron microscopy was used to verify specific binding of the mabs to their respective targets. specific mabs were used to develop an elisa procedure to monitor the deve ...19921393827
studies on anti-b specificity and heterogeneity of some invertebrate hemagglutinins.hemolymph from charybdis japonica and lymantria dispar, and saline extract from eunice kobiensis agglutinated human group b erythrocytes specifically. multiple agglutinins were identified, and independent agglutinins were separated from hemolymph of charybdis japonica (anti-bcj+anti-xcj) and lymantria dispar (anti-bld+rld) by absorption experiments. by the agglutination and agglutination-inhibition experiments with simple sugars, the specificities of the anti-b agglutinins in these invertebrates ...1979494266
[in vivo and in vitro studies of a cytoplasmic polyhedrosis virus isolated from euxoa scandens, riley (noctuidae, agrotinae)(author's transl)].a virus isolated from the larvaes of euxoa scandens (noctuidae, agrotinae) was studied by histological and histochemical techniques, and by electron microscopic observation of virus suspensions and ultrathin sections of infected tissues. our studies demonstrated that the virus in icosahedral in shape, measures 70 nm and belongs to the cytoplasmic polyhedrosis virus group. only the midgut cells of the larvae were found to be infected and the virus particles were localized in inclusion bodies most ...1977336008
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
characterization of baculovirus p10 synthesis using monoclonal antibodies.a series of monoclonal antibodies were produced against virion proteins of the multicapsid nuclear polyhedrosis virus of orgyia pseudotsugata (opmnpv). four of these antibodies reacted with a protein of 14 kd on western blots of electrophoretically separated opmnpv virion proteins. these antibodies were used to identify immunoreactive clones from a lambda gt11 expression library of opmnpv dna. by hybridization of insert dna from the lambda gt11 clones to blots of digests of opmnpv genomic dna, a ...19872442889
inhibition of potassium-gradient-driven phenylalanine uptake in larval lymantria dispar midgut by two bacillus thuringiensis delta-endotoxins correlates with the activity of the toxins as gypsy moth larvicides. 19911661774
p39, a major baculovirus structural protein: immunocytochemical characterization and genetic location.a series of monoclonal antibodies was produced against proteins from polyhedra-derived virions of the multicapsid nuclear polyhedrosis virus of orgyia pseudotsugata (opmnpv). two of these antibodies (214 and 236) reacted with a protein of 39 kda on western blots of electrophoretically separated opmnpv viron proteins derived from both budded and polyhedra-derived virions. this protein appears to be a major component of both bv and pdv. one of the p39 antibodies was used to characterize p39 synthe ...19883059676
physical map and polyhedrin gene sequence of lymantria dispar nuclear polyhedrosis virus.restriction maps of the 166.6-kb genome of lymantria dispar multiply-enveloped nuclear polyhedrosis virus (ldmnpv clone g) were constructed for bamhi, bglii, ecori, ecorv, hindiii and kpni, using cosmid pvk102 and pbluescript vectors. southern hybridizations indicated that the ldmnpv genome contains five dispersed regions of intragenomic sequence homology. the polyhedrin gene of ldmnpv was located within bglii-e and the sequence of the 735-nucleotide (nt) coding region and 678 nt of flanking dna ...19883063616
a simple method for the extraction of baculovirus dna.we have developed a method using lysozyme for dna extraction from baculoviruses using as a model lymantria dispar nuclear polyhedrosis virus (ldnpv) obtained from infected larvae. this method proved to be quick, inexpensive and the extracted dna was successfully used in molecular hybridization experiments.19921597509
characterization of the infection cycle of the orgyia pseudotsugata multicapsid nuclear polyhedrosis virus in lymantria dispar cells.to characterize the infection cycle of the orgyia pseudotsugata multicapsid nuclear polyhedrosis virus in lymantria dispar cells, the time course of dna synthesis and polyhedron production, and the onset and rate of budded virus production were investigated at three different m.o.i. (5, 10 and 100). in addition, the time course of expression of three proteins (gp64, p39 and polyhedrin) representative of three temporal classes of baculovirus genes was also analysed using western blot analysis. dn ...19902273385
conversion of 3-dehydroecdysone by a ketoreductase in post-diapause, pre-hatch eggs of the gypsy moth lymantria dispar.the prothoracic glands (pgs) of lymantria dispar (day-5 female, last-stage larvae) produce both ecdysone and an ecdysteroid which has the same retention time on reverse-phase liquid chromatography (rplc) as a known standard of 3-dehydroecdysone. the latter ecdysteroid can be converted by a heat-labile factor in extracts of post-diapause, pre-hatch l. dispar eggs to an ecdysteroid which has the same retention time on rplc as ecdysone. purified 3-dehydroecdysone, similarly treated with egg extract ...19902134169
nucleotide sequence of lymantria dispar nuclear polyhedrosis virus polyhedrin gene.the polyhedrin gene of the nuclear polyhedrosis virus of the gypsy moth (lymantria dispar) (ldmnpv) was cloned and sequenced. a polyhedrin open reading frame of 735 nucleotides (nt) was identified which can code for a protein of 245 amino acids that demonstrates a high degree of similarity to other polyhedrins. the protein predicted from the nucleotide sequence shows differences in several regions to that previously sequenced from the ldmnpv polyhedrin protein. the consensus sequence aataagtattt ...19892656870
production of recombinant proteins by baculovirus-infected gypsy moth cells.an experimental study was undertaken to evaluate alternative insect cell lines to sf9 [from spodoptera frugiperda (fall armyworm)] for the production of recombinant proteins. insect cell lines from two different organisms were considered: iplb-ldeita (ldeita) from lymantria dispar (gypsy moth) and iplb-hvt1 (hvt1) from heliothis virescens (tobacco budworm). both ldeita and hvt1 produced higher total activity levels of recombinant beta-galactosidase in monolayer culture than sf9 after infection w ...19911367995
nucleotide sequence, transcriptional mapping, and temporal expression of the gene encoding p39, a major structural protein of the multicapsid nuclear polyhedrosis virus of orgyia pseudotsugata.the gene encoding the 39-kda major structural protein (p39) of orgyia pseudotsugata nuclear polyhedrosis virus (opmnpv) was sequenced and transcriptionally mapped, and its expression was examined at various times postinfection. by northern hybridization, primer extension, and s1 nuclease analysis, we identified p39 mrnas of approximately 2600 nt. by primer extension analysis, we identified two major sets of transcripts which initiated around -48 and -96 nt upstream of the translation start codon ...19892644736
identification of essential trans-acting regions required for dna replication of the orgyia pseudotsugata multinucleocapsid nuclear polyhedrosis virus: lef-1 is an essential replication gene.a transient replication assay for the identification of baculovirus genes that are essential for replication of an origin-containing reporter plasmid was established for the orgyia pseudotsugata multinucleocapsid nuclear polyhedrosis virus (opmnpv). using a replication origin located on the opmnpv hindiii-n fragment, we identified a subset of cosmids and plasmids from an opmnpv cosmid library that was able to supply all the essential trans-acting factors and support replication of the origin-con ...19957886946
serological relationship of nuclear polyhedrosis viruses. i. hemagglutination by polyhedral inclusion body protein from the nuclear polyhedrosis virus of porthetria (equal to lymantria) dispar. 19751090680
sequence analysis and transcriptional mapping of the orf-2 gene of autographa californica nuclear polyhedrosis virus.sequencing of the dnapol promoter region of autographa californica nuclear polyhedrosis virus (acnpv) revealed an overlapping open reading frame (orf) in an antisense orientation, referred to as orf-2. analysis of the orf-2 deduced amino-acid sequence revealed two short regions of homology with a similar orf from lymantria dispar nuclear polyhedrosis virus (ldnpv). two 3' processing signals of this gene, expressed late during infection, were shown to be located on the orf-2 stop codon and 162 nu ...19957835700
dna hybridization assay for detection of gypsy moth nuclear polyhedrosis virus in infected gypsy moth (lymantria dispar l.) larvae.radiolabeled lymantria dispar nuclear polyhedrosis virus dna probes were used in a dna hybridization assay to detect the presence of viral dna in extracts from infected larvae. total dna was extracted from larvae, bound to nitrocellulose filters, and assayed for the presence of viral dna by two methods: slot-blot vacuum filtration and whole-larval squashes. to test the assays, neonate larvae were fed droplets containing a known concentration of l. dispar nuclear polyhedrosis virus and observed f ...19892696426
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
expression of pheromone binding proteins during antennal development in the gypsy moth lymantria dispar.we have identified 2 olfactory specific proteins in the gypsy moth lymantria dispar that are uniquely associated with the male antennae, the principal olfactory organs of this animal. these proteins were the major soluble protein components of the olfactory sensilla, present in equivalent amounts. both proteins comigrated on sds-page, showing an apparent molecular mass of 15,000 da but migrated separately on non-sds-page, indicating differences in net charge. n-terminal amino acid sequence analy ...19892795166
cytolytic differences among lepidopteran cell lines exposed to toxins of bacillus thuringiensis subsp. kurstaki (hd-263) and aizawai (hd-112): effect of aminosugars and n-glycosylation.comparison of lytic-dose response behavior of seven lepidopteran cell lines to the activated delta-endotoxin polypeptides of bacillus thuringiensis subspecies kurstaki (hd-263) and aizawai (hd-112) indicated distinct differences among the lines. the lines derived from spodoptera species s. exigua (urc-se-1a) and s. littoralis (uiv-sl-575) were more susceptible to lysis by aizawai toxin (bta) than kurstaki toxin (btk) as were cells from the lymantria dispar line (iplb-ld652y). however, the concen ...19947842169
in vivo isolation of baculovirus genotypes.restriction endonuclease profiles of a wild-type granulosis virus from artogeia rapae (argv3) and a nuclear polyhedrosis virus from lymantria dispar (ldmnpv) indicated that the preparations were genotypically heterogeneous. eight constituent genotypes were isolated from argv3 by low mortality dose infections of late instar a. rapae and analysis of progeny viral dna recovered from individual cadavers. three distinct genotypes were isolated from ldmnpv by the same approach, using l. dispar larvae. ...19882842948
irreversible binding kinetics of bacillus thuringiensis cryia delta-endotoxins to gypsy moth brush border membrane vesicles is directly correlated to toxicity.to examine the binding of bacillus thuringiensis delta-endotoxins, cryiaa, cryiab, and cryiac, to lymantria dispar (gypsy moth) brush border membrane vesicles (bbmv), saturation kinetic analyses were conducted according to a two-step interaction scheme [formula: see text] for delta-endotoxin binding to bbmv, rather than the one-step reversible binding presented in prior reports. the order of toxicity of the delta-endotoxins, as measured by the dose required for a 50% inhibition of weight gain (i ...19957559587
replication of orgyia pseudotsugata baculovirus dna: lef-2 and ie-1 are essential and ie-2, p34, and op-iap are stimulatory genes.a transient dna replication assay was used to identify genes located within m.u. 90.5-7.0 of the orgyia pseudotsugata multinucleocapsid nuclear polyhedrosis virus (opmnpv) genome that influenced replication of a reporter plasmid containing an opmnpv origin of replication, when cotransfected into uninfected lymantria dispar cells. the viral transactivator ie-1 and a 2.4-kb subclone were found to be essential for replication. the 2.4-kb region was sequenced and open reading frames were identified. ...19957571435
characterization, sequencing and phylogeny of the ecdysteroid udp-glucosyltransferase gene from two distinct nuclear polyhedrosis viruses isolated from choristoneura fumiferana.the ecdysteroid udp-glucosyltransferase (egt) gene isolated from a plaque-purified isolate of choristoneura fumiferana multinucleocapsid nuclear polyhedrosis virus (cfmnpv) was compared to its homologue from a defective mnpv virus (cfdef) present in wild type virus populations infecting the eastern spruce budworm, c. fumiferana. the egt genes were located in the same relative position within the virus genomes and their genomic location and arrangement were similar to that found in autographa cal ...19957595348
the effect of inhibitors on the growth of the entomopoxvirus from amsacta moorei in lymantria dispar (gypsy moth) cells.within the family of poxviridae, the entomopoxviruses are the most distant relatives of the more well-known and intensively studied orthopoxviruses (vaccinia and variola). the growth of the entomopoxvirus from amsacta moorei (amepv) has been characterized in cell culture and compared to that of vaccinia virus (vv), the prototypic orthopoxvirus. the overall characteristics of infected cell cultures were generally similar between the two viruses. one striking difference noted was the apparent abse ...19957645250
protein synthesis in cells infected by chilo iridescent virus (iridovirus, type 6).synthesis of the polypeptides induced in civ-infected cells was studied using radiolabelled methionine in a permissive cell line of choristoneura fumiferana. analysis of labelled cell extracts by high resolution polyacrylamide gel electrophoresis (page) revealed the sequential appearance of at least 28 structural and non-structural polypeptides in permissive conditions. this was confirmed by analysis of protein synthesis in non-permissive cell lines of lymantria dispar, aedes albopictus or by th ...19854062562
complete base sequence for the mitochondrial large subunit ribosomal rna of the gypsy moth lymantria dispar (l.).a 1355 bp sequence (accession number l32141) isolated from a gypsy moth (lymantria dispar) cdna library showed 68-74% sequence identity to mitochondrial large subunit ribosomal rna (mt irrna) sequences of locusta migratoria, apis mellifera, aedes albopictus, anopheles gambiae and two drosophila species. a comparison of the primary sequences of the mt irrnas from the above insects in four orders and from esherichia coli demonstrated regions of conservation which presumably correspond to regions o ...19947704305
restriction mapping of lymantria dispar nuclear polyhedrosis virus dna: localization of the polyhedrin gene and identification of four homologous regions.the genome of the multiple-embedded nuclear polyhedrosis virus (mnpv) of lymantria dispar (ldmnpv) was partially characterized by restriction endonuclease analysis and a physical map was constructed using cosmid cloning and southern cross blot hybridization. using bamhi, bglii, ecori and hindiii, the size of the genome was estimated to be 88.5 x 10(6) mr or 134.04 kbp. ldmnpv dna was also analysed using methylation-sensitive restriction enzymes. the resulting restriction profiles suggested that ...19882842437
temporal analysis and spatial mapping of lymantria dispar nuclear polyhedrosis virus transcripts and in vitro translation polypeptides.genomic expression of the lymantria dispar multinucleocapsid nuclear polyhedrosis virus (ldmnpv) was studied. viral specific transcripts expressed in cell culture at various times from 2 through 72 h postinfection were identified and their genomic origins mapped through northern analysis. sixty-five distinct transcripts were identified in this analysis. most viral transcripts were expressed late in infection, and originated from throughout the viral genome. viral polypeptides expressed in infect ...19911685048
[range of the hosts of the causative agent of pébrine (nosema bombycis) in the mulberry silkworm].biological interrelations between the agent of microsporidiosis of the silkworm and local species of insects were studied. 7 species of lepidoptera (agrotis segetum, chloridea obsoleta, laphygma exigua, plusia gamma, pieris brassicae, p. rapae, lymantria dispar) were infected with the suspension of nosema bombycis spores. results have shown that 6 species of insects are susceptible to nosema bombycis excluding only the gipsy moth. so, n. bombycis can be considered to be a parasite with a wide ra ...19806769083
[determination of lysine residue number, positive charge and molecular lengths of histone h1 and h5 by a method of incomplete succinylation].a simple method for determination of lysine residue number and positive charge of histones h1 and h5 in the acetic acid -- urea system has been developed. the method is based on incomplete succinylation of lysine residues and allows an accurate determination of protein molecular length. the accuracy of the method has been demonstrated by determination of goose and chicken histones h5 and their fragments. the calibrating curve for determination of molecular lengths of histones h1 and h5 has been ...19826812653
the baculovirus gp64 envelope fusion protein: synthesis, oligomerization, and processing.the baculovirus gp64 envelope fusion protein (gp64 efp) is a class i integral membrane protein that enters the secretory pathway and is oligomerized and extensively processed during transport to the plasma membrane. the kinetics of gp64 efp biosynthesis, oligomerization, and processing in orgyia pseudotsugata multicapsid nuclear polyhedrosis virus (opmnpv)-infected lymantria dispar cells were examined by pulse label, pulse-chase, and immunoprecipitation experiments. relative rates of gp64 efp sy ...19957778291
field-flow fractionation of alkali-liberated nuclear polyhedrosis virus from gypsy moth lymantria dispar linnaeus.rod-shaped viral particles of the gypsy moth nuclear polyhedrosis virus (npv) were obtained through alkaline dissolution of inclusion bodies isolated and purified from infected hosts. the liberated viral fraction contains a wide assortment of enveloped aggregated forms as well as enveloped monomers. this complex mixture was separated by means of sedimentation field-flow fractionation (sedimentation fff) to give a mass spectrum of the separated particles. from the elution pattern effective molecu ...19807014576
electrophoretic characterization of proteins and rna of cytoplasmic polyhedrosis virus (cpv) from euxoa scandens.a cytoplasmic polyhedrosis virus (cpv) was isolated from euxoa scandens and propagated in vitro in lymantria dispar cells. the virions and polyhedra were analyzed by polyacrylamide gel electrophoresis. polyhedra produced in vivo and in vitro contained six polypeptides with identical relative mobilities while nonoccluded viral particles produced in vivo were composed of four polypeptides. the rna content of the purified virions extracted from infected midgut was resolved in ten segments with mole ...19817036955
identification and characterization of a second putative origin of dna replication in a baculovirus of orgyia pseudotsugata.a 7.5-kb region (96.8-2.5 m.u.), called op5, of the orgyia pseudotsugata multinucleocapsid nuclear polyhedrosis virus (opmnpv) genome that contains an origin of dna replication was characterized. this region replicates several times more efficiently and is unrelated to the previously identified putative origin of replication located on the viral hindiii-n fragment. in contrast to hindiii-n, the origin on op5 contains repeated sequences with limited sequence identity to the homologous regions fro ...19957886962
phylogenetic relationships among vairimorpha and nosema species (microspora) based on ribosomal rna sequence data.a portion (approximately 350 nucleotides) of the large subunit ribosomal rna (rrna) 5' to the 580 region (escherichia coli numbering) was sequenced using the reverse transcriptase dideoxy method and compared for several species of nosema and vairimorpha. comparison among nosema species suggests that this genus is composed of several unrelated groups. the group which includes the type species, nosema bombycis, consists of closely related species found primarily in lepidoptera. other nosema specie ...19947963643
lymantria dispar nuclear polyhedrosis virus homologous regions: characterization of their ability to function as replication origins.homologous regions (hrs) were identified in the lymantria dispar nuclear polyhedrosis virus (ldmnpv) genome. a 1.58-kb region surrounding hr4 was sequenced and found to have two distinct domains. domain i (about 600 bp) is composed of seven repeats of about 80 bp including a series of palindromes containing mlui sites and overlapping xhoi and saci sites. domain ii (about 700 bp) is composed of eight partially repeated sequences of 60 to 100 bp containing a 15- to 25-bp sequence that is 80 to 100 ...19957983712
domain iii exchanges of bacillus thuringiensis cryia toxins affect binding to different gypsy moth midgut receptors.aminopeptidase-n, purified from gypsy moth (lymantria dispar l.) brush border membrane vesicles, exhibited specific binding to cryiac toxin but not to cryiaa toxin. cryiaa-cryiac hybrid toxins were used to localize the aminopeptidase-n binding region on cryiac. slot blot assays and ligand blot experiments demonstrated that the hybrid toxins which have the residues 451 to 623, comprising essentially domain iii, from cryiac toxin exhibited strong binding to purified aminopeptidase-n and 120 kda br ...19957488105
location of a lepidopteran specificity region in insecticidal crystal protein cryiia from bacillus thuringiensis.the bacillus thuringiensis insecticidal crystal protein cryiia has both high mosquito activity and gypsy moth activity; in contrast cryiib, which is 87% homologous, displays no mosquito activity and has a three-fold lower gypsy moth activity. the regions responsible for specificity against gypsy moth (lymantria dispar) and mosquito (aedes aegypti) larvae were located by introducing mlui and xhoi sites into homologous positions within the putative domain ii of both cryiia and cryiib genes, which ...19947997170
insect tolerance of transgenic populus nigra plants transformed with bacillus thuringiensis toxin gene.leaves and stem segments of populus nigra were transformed with a. tumefaciens lba4404 harboring a binary vector containing chimeric genes of npt and 35s-omega-b.t. toxin-nos. nineteen regenerated kanamycin resistant plants were analyzed by dna hybridization, out of which 10 were shown to be the candidates of transgenic plants. insect tolerance tests showed that the transgenic plants were toxic to two lepidopteran pests, lymantria dispar l. and apocheimia cinerarius erschoff. based on the result ...19938061230
identification and characterization of a protein kinase gene in the lymantria dispar multinucleocapsid nuclear polyhedrosis virus.the lymantria dispar multinucleocapsid nuclear polyhedrosis virus (ldmnpv) gene encoding vpk has been cloned and sequenced. ldmnpv vpk shows a 24% amino acid identity to the catalytic domains of the eucaryotic protein kinases npkc from rabbits, hspkce from humans, aplpkcb from aplysia californica, and dpkc98f from drosophila melanogaster, and homology to several other protein kinases from yeasts, mice, and bovines. the homology suggests that vpk is a serine/threonine protein kinase as defined by ...19948107234
identification and characterization of the ecdysteroid udp-glucosyltransferase gene of the lymantria dispar multinucleocapsid nuclear polyhedrosis virus.we have located, cloned, sequenced and characterized the ecdysteroid udp-glycosyltransferase gene (egt) gene from the baculovirus lymantria dispar multinucleocapsid nuclear polyhedrosis virus. (ldmnpv), which is specific for the gypsy moth (l. dispar). the egt gene from the related baculovirus autographa californica multinucleocapsid nuclear polyhedrosis virus (acmnpv) disrupts the hormonal balance of the host larva by galactosylating ecdysone, which prevents moulting. the location of the ldmnpv ...19948151299
[action of streptococcus faecalis on tussock moth larvae (lymantria dispar)]. 19836408898
characterization of productive and non-productive acmnpv infection in selected insect cell lines.we have investigated the process of productive and non-productive acmnpv infection in a variety of cultured insect cells including lines derived from spodoptera frugiperda (iplb-sf21), mamestra brassicae (ses-mabr-3), choristoneura fumiferana (iprl-cf-1), bombyx mori (bmn-4), lymantria dispar (iplb-ld652y), helicoverpa zea (hz1b3), and drosophila melanogaster (dm). in each cell line, we have examined viral utilization of an early, a late, and a very late promoter, the replication of viral dna an ...19938212569
horseradish peroxidase-labelled probes and enhanced chemiluminescence to detect baculoviruses in gypsy moth and eastern spruce budworm larvae.horseradish peroxidase-labelled whole genomic dna probes and enhanced chemiluminescence procedures were utilized to detect baculovirus in insect macerates blotted on nylon membranes. detection levels were similar to those found using 32p-labelled probes; 5 x 10(3) occlusion bodies (obs), 2 x 10(3) obs and 4 x 10(4) obs of lymantria dispar (l.) nuclear polyhedrosis virus (ldnpv), choristoneura fumiferana (clem.) npv (cfnpv) and c. fumiferana granulosis virus (cfgv) respectively using 10 ng/ml ldn ...19938227282
identification of genes encoding late expression factors located between 56.0 and 65.4 map units of the autographa californica nuclear polyhedrosis virus genome.using a previously developed method that allows the identification of autographa californica nuclear polyhedrosis virus (acmnpv) genes which stimulate transient expression from a late and a very late viral promoter [passarelli and miller, j. virol., 67, 2149-2158 (1993)], we have identified three genes between 56.0 and 65.4 map units of the acmnpv genome involved in expression from a late and a very late promoter but not from an early viral promoter. one gene, p143, was previously shown to be es ...19938249293
identification and characterization of a putative origin of dna replication in the genome of a baculovirus pathogenic for orgyia pseudotsugata.a four-kilobase (kb) region (hindiii-n, map units 7.0-11.3) of the orgyia pseudotsugata multinucleocapsid nuclear polyhedrosis virus (opmnpv) genome was found to contain sequences that conferred upon plasmids the ability to undergo infection-dependent replication. the plasmid dna appeared to be replicated to form high molecular weight multimers. plasmids with deletions of up to 1.8 kb from either end of the hindiii-n region were replication competent. however, a discrete sequence, contained with ...19938249294
a 25-kda protein is associated with the envelopes of occluded baculovirus virions.antiserum produced against preoccluded virions of the orgyia pseudotsugata multinucleocapsid nuclear polyhedrosis virus (opmnpv) was used to screen an opmnpv lambda gt11 expression library. one of the immunoreactive clones contained an insert that hybridized to a portion of the opmnpv hindiii-p fragment. a 2-kb region of hindiii-p was sequenced and found to contain three open reading frames, each preceded by a late promoter element. the insert from the lambda gt11 clone was derived from the thir ...19938337828
production of cytoplasmic polyhedrosis virus (cpv) polyhedra in a gamma irradiated lymantria dispar cell line.lymantria dispar cells wee exposed to different doses of gamma radiation one hour after infection with cytoplasmic polyhedrosis virus (cpv). it was found that irradiated cells can produce infectious polyhedra. modifications in the structure and in the process of maturation of the polyhedra were noted. the number of polyhedra per cell increased significantly after cell irradiation at 10(4) and 10(5) rads but no change was noted after cell treatment at 10(2) rads. on the other hand, morphological ...19817023418
toxicity of the venom from nasonia vitripennis (hymenoptera: pteromalidae) toward fly hosts, nontarget insects, different developmental stages, and cultured insect cells.a venom preparation from nasonia vitripennis, a wasp ectoparasitoid of fly pupae, was assayed for lethality in different stages of insects representing ten different orders and in cultured insect cells. in most cases, the motor activity of the injected insects remained completely normal for 1-2 days after the injection and displayed none of the symptoms of paralysis commonly reported for venoms of the hymenoptera. a natural host, the flesh fly sarcophaga bullata, was highly sensitive in the pupa ...19938342173
characterization of an early gene coding for a highly basic 8.9k protein from the orgyia pseudotsugata multicapsid nuclear polyhedrosis virus.a new gene of the baculovirus orgyia pseudotsugata multicapsid nuclear polyhedrosis virus (opmnpv) has been identified that encodes a highly basic 8.9k protein. the gene called p8.9 is expressed as a 0.5 kb transcript by 1 h post-infection and initiates at an early gene motif. the promoter of the 0.5 kb transcript has two upstream elements, repeats i and ii, which are similar to motifs previously characterized in the opmnpv ie-2 gene and the autographa californica nuclear polyhedrosis virus ie-n ...19938345350
a 37-kilodalton glycoprotein from a baculovirus of orgyia pseudotsugata is localized to cytoplasmic inclusion bodies.the gene encoding a 37-kda glycoprotein (gp37) of orgyia pseudotsugata multinucleocapsid nuclear polyhedrosis virus (opmnpv) was located and sequenced. gp37 of opmnpv was found to have 62 and 37% amino acid sequence identity with gp37 of autographa californica multinucleocapsid nuclear polyhedrosis virus (acmnpv) and with a protein reported to be a component of occlusion bodies from choristoneura biennis entomopoxvirus, respectively. the mrna start site of the opmnpv gp37 gene was mapped within ...19938380088
immunocytochemical characterization of p24, a baculovirus capsid-associated protein.an open reading frame (orf 1) located upstream of the polyhedron envelope protein gene in the orgyia pseudotsugata multinucleocapsid nuclear polyhedrosis virus (opmnpv) genome was cloned in-frame into a trpe bacterial expression vector. the fusion protein produced by this construct was used for the preparation of a monospecific antiserum. western blot analysis of extracts from opmnpv-infected lymantria dispar cells and autographa californica npv (acmnpv)-infected spodoptera frugiperda cells dete ...19938423444
saturable attachment sites for polyhedron-derived baculovirus on insect cells and evidence for entry via direct membrane fusion.this research provides the first evidence for specific receptor binding of polyhedron-derived baculovirus (pdv) to host cells and to lepidopteran brush border membrane vesicles (bbmv) and demonstration of entry via a nonendocytotic pathway involving direct membrane fusion. the technique of fluorescence-activated cell sorting analysis was used to investigate the specificity of binding between the pdv phenotype of lymantria dispar nuclear polyhedrosis virus (ldnpv) and host membranes. fluorescein ...19938445715
analysis of the role of 5' promoter elements and 3' flanking sequences on the expression of a baculovirus polyhedron envelope protein gene.the polyhedron envelope protein gene of the orgyia pseudotsugata multinucleocapsid nuclear polyhedrosis virus (opmnpv) is the third in a series of five open reading frames (orfs 1-5) oriented in the same direction. individual mrnas initiate at conserved late gene promoter/mrna start site (a/gtaag) sequences located upstream of each orf and the mrnas co-terminate after the fifth orf. to examine the influence of transcription from upstream promoter elements and the presence of an extensive 3' flan ...19938517021
a baculovirus encoded 16-kda glycoprotein localizes near the nuclear membrane of infected cells.an open reading frame (orf 2) located upstream of the polyhedron envelope protein gene of orgyia pseudotsugata multicapsid nuclear polyhedrosis virus (opmnpv) was cloned in frame into a trpe expression vector. the fusion protein produced by this construct was used for the production of a monospecific antiserum. western blot analysis of opmnpv-infected lymantria dispar cells detected a 16-kda protein at 24 hr postinfection. the 16-kda protein was determined to be n-glycosylated by tunicamycin tre ...19938517029
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
nucleotide sequence and transcriptional analysis of the dna polymerase gene of bombyx mori nuclear polyhedrosis virus.a gene encoding a putative dna polymerase (pol) of bombyx mori nuclear polyhedrosis virus (bmsnpv) was cloned and sequenced. the gene included an open reading frame (orf) encoding a polypeptide of 988 amino acids with a predicted molecular mass of 114.65 kda. the deduced amino acid sequence of the bmsnpv pol orf showed an overall identity of 96 and 45% to those of the autographa californica npv (acmnpv) pol orf and the lymantria dispar npv pol orf, respectively, and contained sequences conserved ...19957831799
brush border membrane aminopeptidase-n in the midgut of the gypsy moth serves as the receptor for the cryia(c) delta-endotoxin of bacillus thuringiensis.aminopeptidase-n (ap-n) was purified from gypsy moth (lymantria dispar, l.) brush border membrane vesicles (bbmv) proteins by mono-q chromatography and superdex-75 gel filtration in the presence of the zwitterionic detergent, chaps, using fplc. the purified ap-n, identified by its enzymatic activity, had an apparent size of 100 kda, and was identified as the unique bacillus thuringiensis insecticidal toxin, cryia(c), binding protein. ap-n clearly displayed strong binding to cryia(c), exhibiting ...19958580914
synergistic effect of the bacillus thuringiensis toxins cryiaa and cryiac on the gypsy moth, lymantria dispar.the insecticidal activity of toxins cryiaa, cryiab, and cryiac against lymantria dispar (gypsy moth) and bombyx mori (silkworm) was examined by force-feeding bioassays. toxin cryiaa exhibited higher toxicity than toxins cryiab and cryiac for l. dispar and b. mori. to evaluate possible synergism among these toxins, bioassays were performed with mixtures of cryiaa and cryiab, cryiab and cryiac, and cryiaa and cryiac. expected toxicity was calculated from the activity of each individual toxin and i ...19968593057
baculovirus replication alters hormone-regulated host development.the baculovirus lymantria dispar nuclear polyhedrosis virus interferes with insect larval development by altering the host's hormonal system. the level of haemolymph ecdysteroids, the insect moulting hormone, was found to be higher in virus-infected larvae than in uninfected controls. this was consistently observed in both fourth instars and day 5-infected fifth instars. the rate of hormone synthesis was examined by in vitro incubation of the prothoracic gland. gland activity in virus-infected l ...19968601794
inconsistencies in determining bacillus thuringiensis toxin binding sites relationship by comparing competition assays with ligand blotting.receptor binding properties of cry1aa, cry1ab, and cry1ac bacillus thuringiensis toxins to lymantria dispar brush border membrane vesicles (bbmv) were investigated by competition assays and bbmv ligand blotting. homologous competition binding assays demonstrated that all cry ia toxins bound to l. dispar bbmv with high binding affinities. heterologous competition assays suggested that all three toxins share the same binding sites. however, our ligand blotting experiments were not consistent with ...19968607806
the dna polymerase and helicase genes of a baculovirus of orgyia pseudosugata.regions of the genome of the orgyia pseudotsugata multinucleocapsid nuclear polyhedrosis virus (opmnpv) containing the dna polymerase and helicase genes were sequenced. the dna polymerase and helicase genes encode predicted proteins of 985 (112.6 kda) and 1223 (140.5 kda) amino acids and exhibited 63 % and 59 % amino acid identity, respectively, with their homologues in the autographa californica mnpv (acmnpv). the influence of sequence variation between the opmnpv and acmnpv dna polymerase and ...19968609478
identification of baculovirus gene that promotes autographa californica nuclear polyhedrosis virus replication in a nonpermissive insect cell line.a gene that promotes autographa californica m nuclear polyhedrosis virus (acmnpv) replication in iplb-ld652y cells, a cell line that is nonpermissive for acmnpv, was identified in lymantria dispar m nuclear polyhedrosis virus (ldmnpv). cotransfection of acmnpv dna and a plasmid carrying the ldmnpv gene into iplb-ld652y cells results in acmnpv replication. the gene maps between 43.3 and 43.8 map units on the 162-kbp genome of ldmnpv. it comprises a 218-codon open reading frame and encodes a polyp ...19968642646
aminopeptidase n purified from gypsy moth brush border membrane vesicles is a specific receptor for bacillus thuringiensis cryiac toxin.we have evaluated the binding of bacillus thuringiensis cry toxins to aminopeptidase n (apn) purified from lymantria dispar (gypsy moth) brush border membrane vesicle (bbmv). cryiac toxin bound strongly to apn, while either the structurally related cryiaa and cryiab toxins or cryic, cryiia, and cryiiia toxins showed weak binding to apn. an in vitro competition binding study demonstrated that the binding of cryiac to l. dispar bbmv was inhibited by apn. inhibition of short circuit current for cry ...19968702277
sex-specific and hormone-controlled expression of a vitellogenin-encoding gene in the gypsy moth.microvitellogenin and vitellogenin cdna from manduca sexta (tobacco hornworm) were tested for use as molecular probes to investigate the expression of genes coding for vitellogenins in spodoptera frugiperda (fall armyworm) and lymantria dispar (gypsy moth). cross-hybridization was not observed between the m. sexta cdnas and s. frugiperda dna and mrna. vitellogenin cdna from m. sexta did not hybridize to l. dispar dna or mrna. however, the 834 bp microvitellogenin cdna from m. sexta hybridized to ...19968742824
characterization of the lymantria dispar nucleopolyhedrovirus 25k fp gene.the lymantria dispar nucleopolyhedrovirus (ldmnpv) gene encoding the 25k fp protein has been cloned and sequenced. the 25k fp gene codes for a 217 amino acid protein with a predicted molecular mass of 24870 da. expression of the 25k fp protein in a rabbit reticulocyte system generated a 27 kda protein, in close agreement with the molecular mass predicted from the nucleotide sequence. the gene is located between 40.3 and 40.8 map units on the viral genome. it is transcribed in a counterclockwise ...19968760443
transformation of gypsy moth (lymantria dispar) cell lines by infection with glyptapanteles indiensis polydnavirus.glyptapanteles indiensis, a species of braconid parasitic wasp, infects its host lymantria dispar (gypsy moth) with a polydnavirus (gipdv) to suppress the host immune system during parasitization. here it is shown that gipdv can infect l. dispar cell lines and that a portion of the gipdv genome is stably maintained in infected cells. results of southern hybridization analyses suggested that this portion of the gipdv genome is integrated into the l. dispar cellular genome. this is the first repor ...19968780687
identification and nucleotide sequence of an ecdysteroid udp-glucosyltransferase gene of spodoptera littoralis multicapsid nuclear polyhedrosis virus.the spodoptera littoralis multicapsid nuclear polyhedrosis virus (slmnpv) is a member of the baculoviridae that shows a distant genetic relationship to the prototype autographa californica mnpv (acmnpv). using an acmnpv gene-specific probe, we identified and mapped an ecdysteroid udp-glucosyltransferase (egt) gene in the genome of slmnpv. sequence determination of a part from the hybridizing dna fragment revealed an open reading frame of 1548 nucleotides that exhibits 38% and 44% identity to the ...19958808334
identification and characterization of an early gene in the lymantria dispar multinucleocapsid nuclear polyhedrosis virus.the lymantria dispar multinucleocapsid nuclear polyhedrosis virus (ldmnpv) gene encoding g22 was cloned and sequenced. the g22 gene codes for a 191 amino acid protein with a predicted m(r) of 22,000. expression of g22 in a rabbit reticulocyte system generated a protein with an m(r) of 24,000, in close agreement with the molecular mass predicted from the nucleotide sequence. g22 is not significantly homologous to any known protein, nor is a g22 homologue present in the autographa californica mnpv ...19958847497
involvement of two amino acid residues in the loop region of bacillus thuringiensis cry1ab toxin in toxicity and binding to lymantria dispar.two amino acids, gly and ser, at positions 282 and 283 in the loop region of domain ii of cry1ab2 toxin are substituted with ala and leu in the cry1ab9-033 toxin. cry1ab2 exhibited about a 10-fold increase in toxicity and a 9-fold increase in binding affinity to lymantria dispar compared to cry1ab9-033. however, these toxins showed similar toxicity and binding affinity to manduca sexta and spodoptera exigua. heterologous competition assays and brush border membrane vesicle (bbmv) ligand blotting ...19968954096
protein engineering of bacillus thuringiensis delta-endotoxin: mutations at domain ii of cryiab enhance receptor affinity and toxicity toward gypsy moth larvae.substitutions or deletions of domain ii loop residues of bacillus thuringiensis delta-endotoxin cryiab were constructed using site-directed mutagenesis techniques to investigate their functional roles in receptor binding and toxicity toward gypsy moth (lymantria dispar). substitution of loop 2 residue n372 with ala or gly (n372a, n372g) increased the toxicity against gypsy moth larvae 8-fold and enhanced binding affinity to gypsy moth midgut brush border membrane vesicles (bbmv) approximately 4- ...19968962052
are odorant-binding proteins involved in odorant discrimination?pheromone-sensitive sensilla trichodea of nine moth species belonging to six families and three superfamilies of lepidoptera were immunolabelled with an antiserum against the pheromone-binding protein of antheraea polyphemus. strong immunolabelling of the sensillum lymph was observed in all long sensilla trichodea of a. polyphemus, a. pernyi (saturniidae), bombyx mori (bombycidae) and manduca sexta (sphingidae). very weak labelling was found with all sensilla trichodea of dendrolimus kikuchii (l ...19968985600
[comparative characteristics of nuclear polyhedrosis virus isolates from gypsy moth lymantria dispar l].eight strains of gypsy moth nuclear polyhedrosis virus (npv) were isolated from dead larvae in kirghizia and tyumen and novosibirsk districts. the strains were characterized in terms of polyhedron morphology, biological activity, virus production, and restriction analysis of dna. the isolated strains did not differ in size or shape of polyhedrons. npv strain nshn-2 was characterized by the highest biological activity (ld50 = 1.2 +/- 0.4) x 10(4) and maximum production: 7.3 x 10(8) pib/larva in c ...19979103045
baculovirus-mediated trans-epithelial transport of proteins in infected caterpillars.baculovirus-mediated abnormal protein trafficking was studied in infected caterpillars by using heterologous proteins. the gene for human complement c1r was expressed in larvae of mamestra brassicae by a recombinant autographa californica nuclear polyhedrosis virus (acmnpv) vector. by following the time-course of recombinant c1r distribution among various tissues, cell types and cell organelles, we concluded that the dominant site of recombinant protein synthesis was the fat body, although some ...19979129671
exploration of receptor binding of bacillus thuringiensis toxins.wild type and mutant toxins of bacillus thuringiensis delta-endotoxins were examined for their binding to midgut brush border membrane vesicles (bbmv). cryiaa, cryiab, and cryiac were examined for their binding to gypsy moth (lymantria dispar) bbmv. the binding of cryiaa and cryiac was directly correlated with their toxicity, while cryiab was observed to have lower binding than expected from its toxicity. the latter observation confirms the observation of wolfersberger (1990). the "rule" of reci ...19958524089
lipofectin increases the specific activity of cypovirus particles for cultured insect cells.cytoplasmic polyhedrosis viruses (cpv) are classified as 14 distinct species (electropherotypes) within the genus cypovirus, family reoviridae. cypovirus research has been limited by a lack of appropriate cell culture systems (for each of these virus species) in which the majority of cells can become productively infected. lipofection increased the infection rate of lymantria dispar 652 cells, by virus particles (derived from polyhedra) of orgyia pseudosugata type 5 cypovirus (op-5 cpv), from 3 ...199910204708
experimental gypsy moth (lymantria dispar) ophthalmia nodosa.ophthalmia nodosa is the nodular granulomatous inflammatory response of ocular tissue to caterpillar hairs. we experimentally simulated this condition by surgically implanting gypsy moth hairs in the rabbit cornea. we were not, however, able to produce ophthalmia nodosa solely by placing cilia in the rabbit cul-de-sac. the experimentally induced inflammation closely resembled human ophthalmia nodosa in both its clinical and histopathologic features.19836342583
molecular analysis of an enhancin gene in the lymantria dispar nuclear polyhedrosis virus.a lymantria dispar nuclear polyhedrosis virus (ldmnpv) gene has been identified that encodes a homolog to the granulovirus (gv) enhancin proteins that are capable of enhancing the infection of other baculoviruses. enhancin genes have been identified and sequenced for three species of gvs but have not been found in any other nuclear polyhedrosis virus to date. the ldmnpv enhancin gene is located between 67.6 and 70.1 kbp on the viral genome. northern and primer extension analyses of viral rnas in ...19979343163
characterization of the replication cycle of the lymantria dispar nuclear polyhedrosis virus.the life cycle of the lymantria dispar nuclear polyhedrosis virus (ldmnpv) was characterized through analysis of budded virus (bv) release, the temporal formation of polyhedra, the temporal transcription pattern of representative early, late, and hyper-expressed late genes, and the onset of dna replication in the ld652y cell line. transcripts from the ldmnpv immediate early gene g22 were detected 4 h post infection (h p.i.). the late and hyper-expressed late p39 capsid and polyhedrin genes were ...19979381798
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