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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
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
construction of chimeric insecticidal proteins between the 130-kda and 135-kda proteins of bacillus thuringiensis subsp. aizawai for analysis of structure-function relationship.eight chimeric insecticidal protein (ip) genes were constructed between the 130-kda and 135-kda ip genes of bacillus thuringiensis subsp. aizawai, and expressed in escherichia coli jm103 cells. the characterization of the produced chimeric ips indicated that the variable region (vr1) in the amino-terminal half of the ips is responsible for the insecticidal activity against larvae of spodoptera litura and plutella xylostella. the carboxy-terminal half of vr1 was important for the formation of the ...19901368534
are b.t.k. plants really safe to eat? 19901369294
phorbol esters increase synthesis of decay-accelerating factor, a phosphatidylinositol-anchored surface protein, in human endothelial cells.a number of cell-surface proteins are anchored by a phosphatidylinositol (pi)-glycan moiety. these proteins can be released by pi-specific phospholipases c (pi-plc). decay-accelerating factor (daf) is such a cell-surface protein that protects cells from inadvertent complement attack by binding to and inactivating c3 and c5 convertases. we have studied the regulation of daf synthesis in human umbilical vein endothelial cells (huvec), a cell that has the highest level of surface daf among those hu ...19901688581
at least one bacillus thuringiensis toxin forms ion-selective pores in membranes. 19901690468
field trial of abate and teknar for simulium (diptera: simuliidae) control in india.simulium breeding was eliminated in a river up to 30 km with 0.1 ppm abate (temephos) for 30 min exposure. teknar (bacillus thuringiensis var. israelensis) at 10 ppm for 10 min exposure resulted in 98.4% and 87.9% reduction up to 20 m in 2 sections of a small stream.19901691272
extra cellular hydrolytic enzyme secretion in bacillus thuringiensis h14 & b. sphaericus & their significance in media design.level of extracellular proteolytic and amylolytic enzyme production was determined to assess the ability of these strains in utilizing complex carbon and nitrogen sources. protease secretion reached maximum at around 12th h of growth in case of all the three b. thuringiensis strains and declined sharply thereafter. in b. sphaericus strains the protease level gradually increased and reached maximum at around 24th h of growth. amylase activity was not detectable in the culture supernatants of b. s ...19901693135
[antigenic structure of endotoxins from bacillus thuringiensis].the proteins of parasporal crystals of 9 strains of entomopathogenic bacteria of the bacillus thuringiensis group of different h-serotypes were studied by different methods. electrophoresis showed that crystals from the bacteria of 1, 3, 4, 5, 8, and 10 serotypes contained 1-2 proteins with a molecular weight of 130-145 kd and an additional fraction with a molecular weight of 55-120 kd. double-radial immunodiffusion revealed that the crystals of all the serotypes studied are immunologically rela ...19901693428
delta-endotoxins form cation-selective channels in planar lipid bilayers.delta-endotoxins cryia(c) and cryiiia, two members of a large family of toxic proteins from bacillus thuringiensis, were each allowed to interact with planar lipid bilayers and were analyzed for their ability to form ion-conducting channels. both of these toxins made clearly resolved channels in the membranes and exhibited several conductance states, which ranged from 200 ps to about 4000 ps (in 300 mm kcl). the channels formed by both toxins were highly cation-selective, but not ideally so. the ...19901694077
rabbit small intestinal trehalase. purification, cdna cloning, expression, and verification of glycosylphosphatidylinositol anchoring.alpha,alpha-trehalase (ec 3.2.1.28), an intrinsic protein of intestinal brush-border membranes, was purified to homogeneity from rabbits. partial amino acid sequences were determined. two degenerate oligonucleotides based on the sequence of a cnbr peptide were employed in a polymerase chain reaction to amplify a 71-base pair fragment of trehalase dna with rabbit intestine cdna as a starting template. this fragment was used as a hybridization probe to isolate full length trehalase clones from a r ...19901697585
isolation of the second bacillus thuringiensis rna polymerase that transcribes from a crystal protein gene promoter.a crystal protein gene of bacillus thuringiensis subsp. kurstaki hd-1-dipel is transcribed in vivo from two overlapping promoters that are activated at different times during sporulation. we reported earlier (k. l. brown and h. r. whiteley, proc. natl. acad. sci. usa 85:4166-4170, 1988) that an rna polymerase containing a sigma subunit with an apparent mr of 35,000 can transcribe in vitro from the promoter utilized from early to midsporulation. we now report the isolation of an rna polymerase co ...19901701426
a translation fusion product of two different insecticidal crystal protein genes of bacillus thuringiensis exhibits an enlarged insecticidal spectrum.two truncated bacillus thuringiensis crystal protein genes, belonging to the classes cryia(b) and cryic and both coding for insecticidal n-terminal fragments of the corresponding crystal proteins, were translationally fused. expression of the gene fusion in escherichia coli showed a biologically active protein with a toxicity spectrum that overlapped those of both contributing crystal proteins.199016348153
interactions between bacillus thuringiensis subsp. israelensis and fathead minnows, pimephales promelas rafinesque, under laboratory conditions.interactions between bacillus thuringiensis subsp. israelensis and fathead minnows, pimephales promelas, were studied in laboratory exposures to two commercial formulations, vectobac-g and mosquito attack. mortality among fatheads exposed to 2.0 x 10 to 6.5 x 10 cfu/ml with both formulations was attributed to severe dissolved oxygen depletion due to formulation ingredients rather than to direct toxicity from the parasporal crystal. no adverse effects were observed at 6.4 x 10 cfu/ml and below. f ...199016348271
insecticidal activity of a bacterial crystal protein expressed by a recombinant baculovirus in insect cells.baculoviruses are insect pathogens with a relatively slow speed of action, and this has limited their use as control agents of insect pests. introduction into baculoviruses of genes which code for proteins interfering specifically with insect metabolism or metamorphosis, such as toxins, hormones, and enzymes, may enhance the pathogenicity of these viruses. the complete insecticidal crystal protein gene cryia(b) of bacillus thuringiensis subsp. aizawai 7.21 was engineered into the nuclear polyhed ...199016348284
limiting an insect infestation of nitrogen-fixing root nodules of the pigeon pea (cajanus cajan) by engineering the expression of an entomocidal gene in its root nodules.a region of dna which determined the production of the insecticidal toxin of bacillus thuringiensis subsp. israelensis was cloned into a derivative of a broad-host-range group incq plasmid vector of gram-negative bacteria. the plasmid which we constructed was transferred by conjugative mobilization into a bradyrhizobium species that nodulates pigeon peas. in this species the construction was maintained stably in the absence of selection and expressed the gene that was installed. experiments in a ...199016348294
the phylloplane as a source of bacillus thuringiensis variants.novel variants of bacillus thuringiensis were isolated from the phylloplane of deciduous and conifer trees as well as of other plants. these isolates displayed a range of toxicity towards trichoplusia ni. immunoblot and toxin protein analysis indicate that these strains included representatives of the three principal b. thuringiensis pathotypes active against larvae of the orders lepidoptera, diptera, and coleoptera. we propose that b. thuringiensis be considered part of the common leaf microflo ...199116348400
specificity of activated cryia proteins from bacillus thuringiensis subsp. kurstaki hd-1 for defoliating forest lepidoptera.the insecticidal activity of the cryia(a), cryia(b), and cryia(c) toxins from bacillus thuringiensis subsp. kurstaki hd-1 was determined in force-feeding experiments with larvae of choristoneura fumiferana, c. occidentalis, c. pinus, lymantria dispar, orgyia leucostigma, malacosoma disstria, and actebia fennica. the toxins were obtained from cloned protoxin genes expressed in escherichia coli. the protoxins were activated with gut juice from bombyx mori larvae. biological activity of the individ ...199116348504
erratum.in the second paragraph of the letter ;;bacillus thuringiensis and pest control'' by marvin k. harris (6 sept., p. 1075), the name of the polyphagus insect pest helicoverpa zea was incorrectly spelled.199117831420
tubular bioreactors: case study of bioreactor performance for industrial production and scientific research.application of bioreactors is dominated by industrial production with the consequence that bioreactors also for scientific purposes are mainly used following an empiric pragmatic approach. for the sake of a breakthrough in biotechnology in general, and especially for advanced process development, a more systematic approach is emphasized here. this methodology in bioreactor performance studies is explained and the meaning clarified in a case study of a new type of tubular bioreactor. the central ...199118597337
assembly and deacetylation of n-acetylglucosaminyl-plasmanylinositol in normal and affected paroxysmal nocturnal hemoglobinuria cells.decay-accelerating factor (daf) is anchored in cell membranes by a glycosyl-plasmanylinositol (gpi) moiety that is transferred to it en bloc in the rough endoplasmic reticulum. to analyze the biochemical reactions involved in preassembly of this structure, a human hematopoietic cell-free system was employed. incubation of cell extracts with udp-[3h]glcnac and butanol partitioning of reaction mixtures yielded two products similar in tlc mobility to intermediates described in trypanosoma brucei. b ...19911708886
early response of cultured lepidopteran cells to exposure to delta-endotoxin from bacillus thuringiensis: involvement of calcium and anionic channels.the role of ion channels in the initial steps following exposure of sf-9 lepidopteran insect cells in culture to the delta-endotoxin cryic from the insecticidal bacterium bacillus thuringiensis was investigated using single ionic channel measurements and microspectrofluorescence of the calcium-sensitive probe fura-2. it was found that: (1) the toxin triggers an immediate rise in intracellular calcium; (2) the surge is due to calcium entering the cells via calcium channels; (3) the toxin recruits ...19911711900
comparative analysis of bacillus anthracis, bacillus cereus, and related species on the basis of reverse transcriptase sequencing of 16s rrna.the primary structures of the 16s rrnas of bacillus anthracis, bacillus cereus, bacillus mycoides, and bacillus thuringiensis were determined by using the reverse transcription-dideoxy sequencing method. all of the strains exhibited very high levels of sequence similarity (greater than 99%) that were consistent with the close relationships shown by previous dna hybridization studies. the sequences of b. anthracis sterne and b. cereus emetic strain nctc 11143 were found to be identical for a cont ...19911715736
potentiation of antitumor activity of bleomycin towards solid tumors in mice by bacillus thuringiensis subsp. israelensis toxin.we previously reported that a 25-kda bacillus thuringiensis subsp. israelensis (bti) toxin is cytotoxic to cultured tumor cells and can also potentiate the cytotoxic effect of some antitumor agents in vitro. in this study, we examined the in vivo effect of bti toxin on the potentiation of antitumor activity of bleomycin (blm) against solid tumor-bearing mice. three tumor cell lines, ehrlich carcinoma, b16 melanoma and meth a fibrosarcoma, were employed. it was shown that dose of bti toxin, ineff ...19911720939
susceptibility of aedes aegypti larvae to temephos and bacillus thuringiensis var israelensis in integrated control.the susceptibility of field collected aedes aegypti larvae was evaluated in terms of median lethal time (lt50) and final mortality, when treated with temephos, bacillus thuringiensis var israelensis as well as mixtures of these two agents. third instar larvae were shown to be more susceptible than early and late fourth instar ones to the entomopathogen. survival of some individuals when exposed to temephos suggest possible resistance. temporal synergism in early fourth instar larvae was detected ...19911726480
[susceptibility of populations of simulium (chirostilbia) pertinax kollar, 1832 (culicomorpha, simuliidae) to eemephos and to bacillus thuringiensis var. israelensis-based formulation].the use of wooden troughs on stream beds, artificially colonized by blackfly larvae, is proposed for larvicide evaluations. mortality was recorded 3 or 4 hours after treatment. larval susceptibility was also evaluated utilizing the lt50 criterion. in there field assays simulium (c.) pertinax populations from the litoral of s. paulo and rio de janeiro states were shown to be resistant to temephos, even when subjected to high concentrations. vectobac 12 as, a bacillus thuringiensis var. israelensi ...19911726481
identification of phosphatidylinositol-specific phospholipase c activity in listeria monocytogenes: a novel type of virulence factor?a phospholipase c which cleaves phosphatidylinositol and glycosylphosphatidylinositol (gpi) anchors was identified in listeria monocytogenes. this 36 kda protein is encoded by the gene plca, and is homologous to the bacillus cereus, bacillus thuringiensis and eukaryotic phosphatidylinositol-specific phospholipases c (pi-plc). expression of the plca gene in escherichia coli correlates with the appearance of pi-plc activity in the cells. in listeria monocytogenes, the activity is secreted to the c ...19911645839
phosphatidylinositol-specific phospholipases c from bacillus cereus and bacillus thuringiensis. 19911646937
is231a from bacillus thuringiensis is functional in escherichia coli: transposition and insertion specificity.a kanamycin resistance gene was introduced within the insertion sequence is231a from bacillus thuringiensis, and transposition of the element was demonstrated in escherichia coli. dna sequencing at the target sites showed that is231a transposition results in direct repeats of variable lengths (10, 11, and 12 bp). these target sequences resemble the terminal inverted repeats of the transposon tn4430, which are the preferred natural insertion sites of is231 in b. thuringiensis.19911648561
sequence of an operon containing a novel delta-endotoxin gene from bacillus thuringiensis.an insect larval toxin designated cryii is produced by several subspecies of bacillus thuringiensis and differs from the other major delta-endotoxins in these bacteria in its size, toxicity profile and presence as part of an operon with three open reading frames (orf). such an operon from a novel b. thuringiensis isolate has been cloned and differs from one previously characterized in the following ways: (a) the size and number of amino acid repeats in one of the orfs; (b) the smaller size of th ...19911651878
a study of human erythrocyte acetylcholinesterase inhibition by chlorpromazine.membrane-bound acetylcholinesterase (ache) from the human erythrocyte is inhibited by chlorpromazine (cpz) in a concentration range within this amphiphilic drug has been demonstrated to interact with erythrocyte membranes, causing a large spectrum of physical and structural effects; membrane solubilization with 0.5% triton x-100 results in a complete loss of cpz inhibitory potency. although these observations might suggest a role of membrane lipid environment in mediating human erythrocyte ache ...19911654884
cloning and analysis of delta-endotoxin genes from bacillus thuringiensis subsp. alesti.bacillus thuringiensis subsp. alesti produced only cryia(b)-type protoxins, and three cryia(b) genes were cloned. one was cryptic because of an alteration near the 5' end, and the other two were very similar to each other. the protoxin encoded by one of the latter genes differed from other cryia(b) protoxins in its greater stability and relative toxicity for two members of the order lepidoptera.19911655719
crystal structure of insecticidal delta-endotoxin from bacillus thuringiensis at 2.5 a resolution.the structure of the delta-endotoxin from bacillus thuringiensis subsp. tenebrionis that is specifically toxic to coleoptera insects (beetle toxin) has been determined at 2.5 a resolution. it comprises three domains which are, from the n- to c-termini, a seven-helix bundle, a three-sheet domain, and a beta sandwich. the core of the molecule encompassing all the domain interfaces is built from conserved sequence segments of the active delta-endotoxins. therefore the structure represents the gener ...19911658659
n-acetyl galactosamine is part of the receptor in insect gut epithelia that recognizes an insecticidal protein from bacillus thuringiensis.proteins synthesized by the bacterium bacillus thuringiensis are potent insecticides. when ingested by susceptible larvae they rapidly lyse epithelial cells lining the midgut. in vitro the toxins lyse certain insect cell lines and show saturable, high-affinity binding to brush-border membrane vesicles (bbmvs) prepared from insect midguts. we observed that the sugar n-acetyl galactosamine (galnac) specifically decreased the cytolytic activity of a cryia(c) toxin towards choristoneura fumiferana c ...19911658805
nucleotide sequence of a novel delta-endotoxin gene cryig of bacillus thuringiensis ssp. galleriae.a gene cryig coding for entomocidal protein delta-endotoxin of bacillus thuringiensis ssp. galleriae str. 11-67 named cryig has been cloned and sequenced (embl accession number x58120). the deduced amino acid sequence that contains 1156 amino acid residues shows only 28% of identical residues, when compared with other delta-endotoxins of the cryi family. the extent of identity is substantially higher for some regions of the sequence ('conserved blocks'), that presumably bear important structural ...19911660003
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
identification and characterization of heliothis virescens midgut membrane proteins binding bacillus thuringiensis delta-endotoxins.to investigate the specificity of bacillus thuringiensis var. kurstaki strain hd1 insecticidal crystal proteins (icp), we used membrane preparations obtained from the midgut of heliothis virescens larvae to perform separate ligand-blot experiments with the three activated cryia toxins. the cryia(a) and the cryia(b) toxins bind the same 170-kda protein, but most likely at two different binding sites. the cryia(c) toxin binds two proteins of molecular masses 140 kda and 120 kda. we also demonstrat ...19911662137
construction of cloning vectors for bacillus thuringiensis.the replication region of the bacillus thuringiensis plasmid, pht1030, was treated with hydroxylamine. various copy-number mutants were selected and subsequently used to construct shuttle vectors with multiple cloning sites. these recombinant plasmids are very stable and allowed the cloning of a delta-endotoxin-encoding gene in b. thuringiensis. comparison between gene expression level and vector copy-number indicated that a plateau in delta-endotoxin production is reached with a copy-number of ...19911662180
differential inhibition by bacillus thuringiensis delta endotoxin of leucine and aspartic acid uptake into bbmv from midgut of manduca sexta.pre-incubation of brush border membrane vesicles (bbmv) isolated from the midgut of manduca sexta with activated bacillus thuringiensis delta endotoxin for a short period resulted in differential inhibition of k(+)-dependent transport of leucine relative to the effect on k(+)-dependent transport of aspartic acid. the difference in i1/2 (5 fold greater for aspartic acid than for leucine) is interpreted as the result of enhanced binding of the b. thuringiensis delta endotoxin to the leucine transp ...19911662492
insect resistance of transgenic tobacco plants expressing delta-endotoxin gene of bacillus thuringiensis.the initiative b.thuringiensis delta-endotoxin (bt toxin) gene clones th12 and th48 contain two different classes of homologous genes, the 5.3 kb class and 6.6 kb class, respectively. bt toxin genes of both classes, modified at the 5'-end and truncated at the 3'-end, can still be expressed to produce the insecticidal, truncated toxin proteins in e. coli. the modified bt toxin genes were inserted into the plant binary expression vector pbin 437 (a derivative plasmid of pbin 19) and were transferr ...19911663400
the c-terminal domain of the toxic fragment of a bacillus thuringiensis crystal protein determines receptor binding.the insecticidal crystal proteins of bacillus thuringiensis show a high degree of specificity. in vitro binding studies with several crystal proteins demonstrated a correlation between toxicity and binding to receptors of larval midgut epithelial cells. in order to study the domain-function relationships of the toxic fragment, hybrid crystal proteins based on cryia(b) and cryic were constructed. two out of 11 hybrid proteins constructed exhibited insecticidal activity. both dispalyed an insectic ...19911664021
stability of the delta-endotoxin gene from bacillus thuringiensis subsp. kurstaki in a recombinant strain of clavibacter xyli subsp. cynodontis.deletion of chromosomally inserted gene sequences from clavibacter xyli subsp. cynodontis, a xylem-inhabiting endophyte, was studied in vitro and in planta. we found that nonreplicating plasmid pcg610, which conferred resistance to kanamycin and tetracycline and contained segments of c. xyli subsp. cynodontis genomic dna, integrated into a homologous sequence in the bacterial chromosome. in addition, pcg610 contains two copies of the gene encoding the cryia(c) insecticidal protein of bacillus th ...19911664710
[search for producers of restriction endonucleases with a given specificity].six site-specific restriction endonucleases were isolated from bacillus thuringiensis strains chosen of 58 strains ought purposefully for production of restriction enzymes. all six strains produce isoschisomers of sau3a.19911667028
fed-batch fermentation studies with bacillus thuringiensis h-14 synthesising endotoxin.cell yield and toxicity of b. thuringiensis h-14 was improved markedly by adopting a simple fed-batch fermentation technique based on controlling glucose concentration. maintenance of steady glucose concentration (0.3-0.5%) in the culture medium was achieved by the continuous addition of concentrated glucose solution. addition of glucose at 3 g/hr/l of culture starting from 3rd hr till 16th hr of fermentation was found to yield cell densities of 80 g/l (wet weight) which represented a nearly 3-f ...19911667779
high-level expression in escherichia coli and rapid purification of phosphatidylinositol-specific phospholipase c from bacillus cereus and bacillus thuringiensis.the construction of four vectors for high-level expression in escherichia coli of the phosphatidylinositol-specific phospholipase c from bacillus cereus or bacillus thuringiensis is described. in all constructs the coding sequence for the mature phospholipase is precisely fused to the e. coli heat-stable enterotoxin ii signal sequence for targeting of the protein to the periplasm. in one set of plasmids expression of the b. cereus or b. thuringiensis enzyme is under control of the e. coli alkali ...19911668269
[bacillus thuringiensis: biological characteristics and production perspectives].in this paper the potential activity of bacillus thuringiensis as insecticide is reviewed. it is presented recent knowledges of its metabolism, production system and the rinetic constants already reported finally the possibilities for production in batches or continuous culture is discussed in order to improve the productivity and economical aspects.19911670264
deletion by in vivo recombination shows that the 28-kilodalton cytolytic polypeptide from bacillus thuringiensis subsp. israelensis is not essential for mosquitocidal activity.the cyta gene encoding the 28-kda polypeptide of bacillus thuringiensis subsp. israelensis crystals was disrupted in the 72-mda resident plasmid by in vivo recombination, thus indicating that homologous recombination occurs in b. thuringiensis. the absence of the 28-kda protein in b. thuringiensis did not affect the crystallization of the other toxic components of the parasporal body (68-, 125-, and 135-kda polypeptides). the absence of the 28-kda protein abolished the hemolytic activity of b. t ...19911675212
effect of corn-steep liquor on growth and mosquito larvicidal activity of bacillus thuringiensis var israelensis de barjac 1978 and b. sphaericus neide 1904.influence of corn steep liquor on the cell yield and toxicity of three strains of b. thuringiensis var israelensis and two strains of b. sphaericus was studied and compared with peptone-yeast extract using a laboratory fermentor. large increase in the cell yield of all the three strains of b. thuringiensis var israelensis was observed when cornsteep liquor was used as the sole nitrogen source. significant increase in toxicity was also observed in b. thuringiensis var israelensis strains b17 and ...19911678365
bacillus sphaericus as a mosquito pathogen: properties of the organism and its toxins.in the course of sporulation, bacillus sphaericus produces an inclusion body which is toxic to a variety of mosquito larvae. in this review we discuss the general biology of this species and concentrate on the genetics and physiology of toxin production and its processing in the midgut of the larval host. the larvicide of b. sphaericus is unique in that it consists of two proteins of 51 and 42 kda, both of which are required for toxicity to mosquito larvae. there is a low level of sequence simil ...19911682792
subspecies-specific haemagglutination patterns of fimbriated bacillus thuringiensis spores.electron microscopy revealed fimbrial appendages (pili) of three types on the surface of bacillus thuringiensis spores. fimbriated spores induced mannose-resistant agglutination of rat, guinea pig and/or chicken red blood cells. each haemagglutination pattern correlated with the subspecies of a given b. thuringiensis strain, rather than with its flagellar serotype. fimbriation and haemagglutination patterns of b. thuringiensis spores are considered to be of possible taxonomic value.19911686005
[mobilization transfer of the pub110 plasmid between gram-positive bacteria].the three factor crosses between the donor strain bacillus subtilis 168 harbouring the plasmid pub102-4, bacillus thuringiensis strain carrying the mobilizing plasmid pam beta 1 and recipient strain lactobacillus fermenti were conducted in order to elaborate the optimal conditions of the plasmid pub102-4 mobilization for transfer into gram-positive microorganisms and to elucidate the possible expression of endogluconase genes in a lactobacillus strain. the lactobacillus fermenti transconjugants ...19911745269
identification of putative insect brush border membrane-binding molecules specific to bacillus thuringiensis delta-endotoxin by protein blot analysis.binding sites for insecticidal toxins of bacillus thuringiensis are located in the brush border membranes of insect midguts. two approaches were used to investigate the interactions of b. thuringiensis subsp. kurstaki hd-73 cryia(c) toxin with brush border membrane vesicles from sensitive and naturally resistant insects: 125i-toxin-vesicle binding assays and protein blots probed with 125i-cryia(c) toxin. in bioassays, manduca sexta and heliothis virescens larvae were highly sensitive, helicoverp ...19911746942
biological calcium absorption edge imaging using monochromatic synchrotron radiation.soft x-ray contact absorption edge images of unfixed, unstained biological specimens were made using monochromatic synchrotron radiation. x-ray contact replicas of unfixed, hydrated biological specimens at the nitrogen absorption edge and above and below the caliii absorption edge were compared to comparative conventional morphological and elemental high-resolution imaging methods (scanning and transmission electron microscopy, tem-histochemistry and tem-x-ray microanalysis). soft x-ray absorpti ...19911755114
toxicity of bacillus thuringiensis to laboratory populations of the olive fruit fly (dacus oleae).a survey of bacillus thuringiensis recovered from the environments of olive groves in greece was carried out. of 80 soil samples, 24 were found to contain b. thuringiensis with parasporal crystal inclusions; these were tested for toxicity against the olive fruit fly (dacus oleae). mortality levels of larvae caused by the different isolates varied from 7 to 87%. higher levels of mortality were observed if a mixture of relatively pure crystals and spores was used compared with the mortality result ...19911768100
cytotoxicity of a cloned bacillus thuringiensis subsp. israelensis cryivb toxin to an aedes aegypti cell line.a cloned cryivb toxin was purified from a cured strain of bacillus thuringiensis (bt) containing the cryivb gene on the recombinant plasmid cam135. solubilized protoxin was treated with aedes gut extract or trypsin for varying times and tested for toxicity in vitro on three dipteran and one lepidopteran cell line. treatment with the aedes extract but not trypsin, produced an active toxin which lysed only aedes aegypti cells out of those tested. this activation was time-dependent reaching a maxim ...19911769533
production & formulation of bacillus thuringiensis var. israelensis & b. sphaericus 1593.three fermentation media each for bulk growth of b. thuringiensis var. israelensis and b. sphaericus 1593 were formulated using defatted groundnut cake (arachis hypogea) as the first nitrogen source and gram flour (cicer arientinum), soy bean (glycine max) and defatted milk powder as the second nitrogen source. medium containing gram flour showed highest toxicity (14.45 micrograms/l) in case of b. thuringiensis var. israelensis whereas medium containing milk powder was found to be highly toxic w ...19911778620
[the flotation characteristics of bacillus cells and spores].variations in hydrophobicity of the surface of bacillary cells and their capacity to flotation in the process of batch cultivation have been studied. it is shown that hydrophobicity of the cell surface increases in the course of batch cultivation of bacillus thuringiensis, b. licheniformis and b. megaterium. hydrophobicity of spores of the mentioned cultures is considerably higher than that of the vegetative cells. the increase of hydrophobicity of bacillary cells positively correlated with thei ...19911779906
prediction of insecticidal activity of bacillus thuringiensis strains by polymerase chain reaction product profiles.a rapid analysis of bacillus thuringiensis strains predictive of insecticidal activity was established by using polymerase chain reaction (pcr) technology. primers specific to regions of high homology within genes encoding three major classes of b. thuringiensis crystal proteins were used to generate a pcr product profile characteristic of each insecticidal class. predictions of insecticidal activity were made on the basis of the electrophoretic patterns of the pcr products. included in the scre ...19911781673
two novel strains of bacillus thuringiensis toxic to coleopterans.two novel strains of bacillus thuringiensis were isolated from native habitats by the use of genes coding for proteins toxic to coleopterans (cryiii genes) as hybridization probes. strain eg2838 (isolated by the use of the cryiiia probe) contained a cryiiia-hybridizing plasmid of approximately 100 mda and synthesized crystal proteins of approximately 200 (doublet), 74, 70, 32, and 28 kda. strain eg4961 (isolated by the use of a cryiiia-related probe) contained a cryiiia-hybridizing plasmid of ap ...19911781691
bioencapsulation and delivery to mosquito larvae of bacillus thuringiensis h14 toxicity by tetrahymena pyriformis. 19911787330
frass failure and pupation failure as quantal measurements of bacillus thuringiensis toxicity to lepidoptera. 19911787331
cloning and expression of the lepidopteran toxin produced by bacillus thuringiensis var sotto in escherichia coli.during sporulation, bacillus thuringiensis var. sotto produces a parasporal crystalline protein which is toxic for the silk-worm, bombyx mori and the cotton leaf worm, spodoptera littoralis. the gene coding this crystal protein is present in a single plasmid. the plasmid dna was isolated, purified and physically mapped using restriction endonuclease enzymes (r.e.). the gene coding the delta-endotoxin was inserted into escherichia coli-jm103, using cloning vector puc8. transformed e. coli cells w ...19911791407
delayed mortality and morphogenetic anomalies induced by the microbial control agent bacillus thuringiensis ser. (h-14) in culex quinquefasciatus.sublethal concentrations of bacillus thuringiensis ser. h-14 were applied to early 4th-instar larvae of culex quinquefasciatus to assess mortality and morphogenetic aberrations in larvae, pupae and adults. at the 24 h lc10, lc25, lc50 and lc80, additional mortality occurred in surviving larvae beyond a 24 h exposure period. the cumulative mortalities increased daily and the overall mortality of larvae up to 7 days posttreatment were 12, 73, 82 and 96% at the indicated concentrations, respectivel ...19911791452
[the autoselection of neustonic forms of bacteria].self-breeding of neuston forms of methylobacterium sp., pseudomonas putida bc-2, alcaligenes paradoxus bc-1, bacillus thuringiensis var. israilensis bacteria as well as of a mixed culture of methylotrophs is shown possible. in spite of ability of hydrophobicity of the cell surface the suggested method of self-breeding may be used to perfect properties of larvicidal biopreparations, and bacterial preparations which intensify self-purification of water bodies.19911791780
[a comparison of some malarial mosquito species in their reaction to bacterial insecticides].considerable differences have been detected in the sensitivity of various anopheles species to bacterial insecticides. these differences might be associated with features in larval nutrition and do not depend on their size and taxonomic species position. an. pulcherrimus or an. sacharovi and an. stephensi species are suggested as the most suitable test objects for the representatives of the given genus.19911795691
action of phosphatidylinositol-specific phospholipase c from bacillus thuringiensis is significantly influenced by coexisting lipids in substrate-detergent micelles.the effects of phosphatidylcholine (pc), phosphatidylethanolamine (pe), sphingomyelin (sm), and cholesterol on the activity of phosphatidylinositol-specific phospholipase c (pi-plc) from bacillus thuringiensis were studied in detail in phosphatidylinositol (pi)/detergent mixed micelles. by addition of pc, the enzymatic hydrolysis of pi was significantly stimulated in pi/triton x-100 as well as pi/sodium deoxycholate (sdc) mixed micelles. sm stimulated enzyme activity toward pi/triton x-100 micel ...19911797426
the effects of coexisting lipids on the action of bacillus thuringiensis phosphatidylinositol-specific phospholipase c toward liposomal substrate.the hydrolytic activity of phosphatidylinositol (pi)-specific phospholipase c (pi-plc) from bacillus thuringiensis was studied in detail toward mixed liposomes consisting of pi and one of other phospholipids and cholesterol. among pi-liposomes, small unilamellar vesicles (suv) were the most sensitive to pi-plc; the enzymatic hydrolysis of pi in suv was not less than 10-fold that in large unilamellar vesicles (luv) or in multilamellar vesicles (mlv). thus, in a survey of the effects of coexisting ...19911799944
long-term effects of bacillus thuringiensis subsp. israelensis on aedes aegypti.immediate (24 hours exposure) and long-term (until the emergence of the adults) effects of different doses of a primary powder of bacillus thuringiensis subsp. israelensis (b.t.i.) against first and second instar aedes aegypti larvae were monitored. the long-term effect was dose-dependent and was materialized by a prolongation of the preimaginal development and continuous cumulative mortality until the emergence of the adults. mortality values were higher during the fourth larval instar and the ...19911802053
isolation and cloning of bacillus thuringiensis var kurstaki hd73 toxin gene and construction of a chimaeric gene for expression in plants.necessity for the production of transgenic crop plants of india, capable of expression of insecticidal bt protein in plant to combat lepidopteran pest damage has been strongly felt. bacillus thuringiensis kurstaki hd73 crystal protein coded by cryia(c)73 gene has been found to be sufficiently effective against the major pests of jute and chickpea. an attempt to isolate the gene and make its use in a chimaeric gene construct for expression in plant was carried out. the plasmid cryia(c)73 gene was ...19911816076
laboratory evaluation of bacillus thuringiensis h-14 against aedes aegypti larvae in the northeast region of thailand.laboratory bioassays using a preparation of bacillus thuringiensis h-14 (bt.h-14), namely skeetal were conducted to determine their effectiveness against late 3rd/early 4th instar larvae of aedes aegypti. the larvae were collected from municipal areas in 7 provinces, namely burirum, roi-et, khon kaen, ubol ratchatani, nakorn phanom, surin and nakorn ratchasima, in the northeast of thailand. it was found that for skeetal, lc50 ranged from 128 to 151 nl/l (average 143) and lc90 ranged from 254 to ...19911818396
[insecticidal activity of various strains of bacillus against larvae of houseflies (musca domestica)].insecticidal activity of b. thuringiensis subsp. morrisoni and b. thuringiensis subsp. darmstadiensis as well as b. sphaericus 2362 and 2297 was evaluated against larvae of the house fly (musca domestica) under laboratory condition. the activity of all tested strains was similar, lc50 ranges from 4 x 10(7) to 1 x 10(8) spores/ml. the higher spores concentration (2 x 10(9)) caused ca 70 per cent mortality. the relative low sensitivity of house fly larvae could results from weakly alkalic ph of th ...19911822051
[usefulness of thuridan in the control of fly larvae in animal houses].the studies were carried out in a pig farm and in a dairy cattle farm in south poland. thuridan was applied as 0.3% suspension in 3 replications in 7 days interval. the efficacy of thuridan was established using fly--catchers which were hung on the same places--once before and three times after thuridan was used. in the animal houses 86% less flies were found.19911823488
aminoacylase i is not a glycolipid-anchored ectoenzyme in pig kidney.subcellular fractionation of pig kidney cortex revealed that aminoacylase i (ec 3.5.1.14, n-acyl-l-amino-acid aminohydrolase) is predominantly a soluble enzyme with only 0.5% of the total activity being recovered in the membrane fraction. the aminoacylase i activity associated with the membrane preparations displayed neither rapid release following incubation with phosphatidylinositol-specific phospholipase c from bacillus thuringiensis nor the distinctive differential pattern of detergent solub ...19911825788
cloning of a tumor-associated antigen: mov18 and mov19 antibodies recognize a folate-binding protein.monoclonal antibodies mov18 and mov19, raised against a membrane preparation of an ovarian carcinoma surgical specimen, react with a surface antigen present on the majority of nonmucinous ovarian malignant tumors tested but not with normal adult tissue (s. miotti, s. canevari, s. ménard, d. mezzanzanica, g. porro, s. m. pupa, m. regazzoni, e. tagliabue, and m. i. colnaghi, int. j. cancer, 39: 297-303, 1987). this surface antigen was purified as a soluble glycoprotein (molecular mass, 36-38 kda) ...19911840502
expression of the cryib crystal protein gene of bacillus thuringiensis.the cryib gene of bacillus thuringiensis subsp. thuringiensis hd-2 codes for a mr 139492 protein that is lethal to certain lepidopteran larvae. we used primer extension to map transcriptional initiation sites and found that cryib was transcribed from two sites that are activated at different times during sporulation. the presumed promoter regions for the two start sites are very similar to the two promoters preceding the cryia (a) gene, and the in vivo transcriptional start sites were found to b ...19911840641
plasmid vectors for 'unclonable' dna constructions. 19911841362
[implications of biological pest and parasite control for environment].biological control of parasites, vectors and pests must consider safety to non-target organisms and to the environment. the who safety procedure was discussed in respect to such agents as bacillus thuringiensis, romanomermis culicivorax, gambusia affinis and others. monitoring of the environment in which biological control treatments are performed is necessary to secure safety requirements as well as the efficiency of biological control programmes.19911844787
mechanism of inhibition of adenylate cyclase by phospholipase c-catalyzed hydrolysis of phosphatidylcholine. involvement of a pertussis toxin-sensitive g protein and protein kinase c.the phospholipase c-mediated hydrolysis of phosphatidylcholine has been shown recently to be activated by a number of agonists. muscarinic receptors, which trigger various signal transduction mechanisms including inhibition of adenylate cyclase through gi, have been shown to be potent stimulants of this novel phospholipid degradative pathway. we demonstrate here, by exogenous addition of bacillus cereus phosphatidylcholine-hydrolyzing phospholipase c, that phosphatidylcholine breakdown mimics th ...19911845988
the mechanism of sunlight-mediated inactivation of bacillus thuringiensis crystals.detailed photostability studies were carried out using purified delta-endotoxin crystals from bacillus thuringiensis subspecies hd-1 and hd-73. the mechanism and time course of sunlight inactivation was investigated by: (a) monitoring the tryptophan damage in the intact crystals by raman spectroscopy, (b) amino acid analysis and (c) biological assays using insects. the results demonstrate that, for purified hd-1 or hd-73 crystals, the 300-380 nm range of the solar spectrum is largely responsible ...19911846530
release of a membrane surface glycoprotein from human platelets by phosphatidylinositol specific phospholipase(s) c.phosphatidylinositol (pi) specific phospholipase c (piase c) treatment of human platelets caused release of a surface glycoprotein in the medium. human blood platelets were isolated by low speed centrifugation and surface glycoproteins were labelled with periodate/[3h]borohydride procedure. intact surface-labelled platelets were treated with piase c purified from culture filtrates of staphylococcus aureus (sa) or bacillus thuringiensis (bt). after piase c treatments platelets were spun at low sp ...19911847299
listeria monocytogenes mutants lacking phosphatidylinositol-specific phospholipase c are avirulent.a number of bacterial species secrete phosphatidylinositol-specific phospholipase c (pi-plc). in this report, we show that the facultative intracellular bacterial pathogen, listeria monocytogenes, contains a gene, plca, predicting a polypeptide with 31% amino acid identity to a bacillus thuringiensis pi-plc. accordingly, l. monocytogenes secretes pi-plc activity, while a mutant with a transposon insertion in plca lacks detectable pi-plc activity. in addition, expression of plca in b. subtilis re ...19911847723
two structural domains as a general fold of the toxic fragment of the bacillus thuringiensis delta-endotoxins.the unfolding by guanidine hydrochloride of the toxic fragment of a bacillus thuringiensis toxin belonging to the cryic class reveals a two-step denaturation under both acid and alkaline conditions. this demonstrates the existence of two structural domains as building blocks for this toxin. protease digests performed on a cryia(b) and cryic b. thuringiensis toxin, under native and partially denatured conditions, confirm this conclusion. whereas the native cryic toxin is completely protease resis ...19911847865
phosphatidylinositol-specific phospholipase c induces biosynthesis of acetylcholinesterase via diacylglycerol in schistosoma mansoni.we have previously shown that two ectoenzymes, acetylcholinesterase (ache) and alkaline phosphatase, are released from the surface and from particulate fractions of the parasite schistosoma mansoni, by a phosphatidylinositol-specific phospholipase c (ptdins-plc) of bacterial origin. exposure to ptdins-plc not only removes large amounts of ache from the surface of intact, viable schistosoma in culture, but is accompanied by a concomitant increase in overall levels of ache in the parasite. the sam ...19911847873
rapid analysis of glycolipid anchors in amphiphilic dimers of acetylcholinesterases.1. we describe two simple procedures for the rapid identification of certain structural features of glycolipid anchors in acetylcholinesterases (aches). 2. treatment with alkaline hydroxylamine (that cleaves ester-linked acyl chains but not ether-linked alkyl chains) converts molecules possessing a diacylglycerol, but not those with an alkylacylglycerol, into hydrophilic derivatives. aches in human and bovine erythrocytes possess an alkylacylglycerol (roberts et al., j. biol. chem. 263:18766-187 ...19911849455
cloning and expression of bacillus thuringiensis israelensis delta-endotoxin dna in b. sphaericus.bacillus thuringiensis israelensis delta-endotoxin genes were cloned into bacillus sphaericus 2362, producing stable transformants reacting with antibody to the 28- and 65-kda b. thuringiensis israelensis crystal proteins and approximately 10 times more toxic to aedes mosquito larvae than the original host strain. the lc50 after 48 hr of exposure of aedes larvae to the most active transformed clone was 0.19 microgram/ml, compared with an lc50 of 1.9 microgram/ml for b. sphaericus 2362 and less t ...19911850771
isolation of mosquito-pathogenic bacillus sphaericus & b. thuringiensis from the root surface of hydrophytes.attempts were made to isolate b. sphaericus and b. thuringiensis active against mosquito larvae from the root surface of hydrophytes. out of 139 samples processed, 86 b. sphaericus and 23 b. thuringiensis isolates were obtained. sixty two of the b. sphaericus isolates belonged to the serotype h5a5b, 2 to h6 and 22 isolates did not agglutinate with any of the 6 antisera tested. twenty of the b. thuringiensis isolates belonged to the h14 serotype, 1 each to the h10 and h17 serotype(s) and 1 to an ...19911855819
characterization and sequence analysis of a small plasmid from bacillus thuringiensis var. kurstaki strain hd1-dipel.the complete nucleotide sequence of a small (2.055 kb) plasmid phd2 from bacillus thuringiensis var. kurstaki strain hd1-dipel was obtained. the sequence encoded two open reading frames (orfs) which corresponded to polypeptides of mr 26,447 and 9122. comparison of the sequence with those obtained for other plasmids from gram-positive organisms suggested that phd2 may belong to the extensive and highly interrelated family of plasmids exhibiting replication via a ssdna intermediate: a putative nic ...19911857751
analysis of unstable rna transcripts of insecticidal crystal protein genes of bacillus thuringiensis in transgenic plants and electroporated protoplasts.we have examined expression of several insecticidal crystal protein (icp) genes of bacillus thuringiensis in transgenic tobacco plants and electroporated carrot protoplasts. we determined that low levels of lepidopteran toxin cryia(b) icp gene expression in plants and electroporated carrot cells is due to rna instability. we used a series of 3' deleted by cryia(b) constructs directed by the cauliflower mosaic virus 35s promoter to demonstrate that this instability is minimally contained in the f ...19911863758
shuttle vectors containing a multiple cloning site and a lacz alpha gene for conjugal transfer of dna from escherichia coli to gram-positive bacteria.the mobilizable shuttle cloning vectors, pat18 and pat19, are composed of: (i) the replication origins of puc and of the broad-host-range enterococcal plasmid pam beta 1; (ii) an erythromycin-resistance-encoding gene expressed in gram- and gram+ bacteria; (iii) the transfer origin of the incp plasmid rk2; and (iv) the multiple cloning site and the lacz alpha reporter gene of puc18 (pat18) and puc19 (pat19). these 6.6-kb plasmids contain ten unique cloning sites that allow screening of derivative ...19911864514
[inversion polymorphism in malaria mosquito anopheles messeae. part x. resistance of larvae with different genotypes to toxins of crystal-forming bacteria bacillus thuringiensis subsp. israelensis (serovar h14)].insensibility of larvae with different chromosomal inversions to the toxins of bacillus thuringiensis subsp. israelensis (bti) was examined. it has been shown that larvae with inversion combinations xl0(xl1)2r0-3r0-3l0 had greater variability after treatment with bti than larvae with inversions xl1(xl2)2r1-3r1-3l1(3l0). the former inversion combinations were previously shown to dominate in the south of the species area and to be supported by k-selection. the latter inversion combinations form "n ...19911874433
effect of certain formulations of the bacterial larvicide, bacillus thuringiensis, serotype h-14 on culex pipiens l. in egypt. 19911875070
effect of nutritive elements on the extracellular protein of different bacillus strains, toxic to mosquito larvae.the effect of 5 nutritional elements (glucose, peptone, yeast extract, beef extract and lactic acid) on the growth of the 4 tested bacillus strains (bacillus thuringiensis bactimos 78, b. thuringiensis ips 82, b. sphaericus solvay 85 and b. sphaericus rb 80) as well as on the concentration of their extracellular proteins were studied. the nutritive elements were individually tested either dissolved in distilled water or in a buffer base. results indicated that the bacterial growth was higher whe ...19911875084
characterization and comparative sequence analysis of replication origins from three large bacillus thuringiensis plasmids.the replication origins of three large bacillus thuringiensis plasmids, derived from b. thuringiensis hd263 subsp. kurstaki, have been cloned in escherichia coli and sequenced. the replication origins, designated ori 43, ori 44, and ori 60, were isolated from plasmids of 43, 44, and 60 mda, respectively. each cloned replication origin exhibits incompatibility with the resident b. thuringiensis plasmid from which it was derived. recombinant plasmids containing the three replication origins varied ...19911885511
two new isolates of bacillus thuringiensis pathogenic to spodoptera litura.both the standard bacillus thuringiensis kurstaki (hd-1) and the formulated commercial product resulted from this strain have shown limited pathogenicity against the tobacco cutworm (spodoptera litura). however, two new isolates of bacillus thuringiensis (k-2074 and k-2178) isolated from taiwan have been identified through an active screening program to be highly pathogenic against the tobacco cutworm. in this paper, we present results of characterization and the pathogenicity of these two new i ...19911885923
viral and bacterial pathogens of insects. 19911887272
field trials with vectolex (bacillus sphaericus) and vectobac (bacillus thuringiensis (h-14)) against anopheles gambiae and culex quinquefasciatus breeding in zaire.under field conditions in kinshasa, zaire, an aqueous suspension of bacillus thuringiensis (h-14), vectobac (12-as), lost most of its larvicidal activity at all concentrations after 48 h against culex quinquefasciatus breeding in polluted gutter water and anopheles gambiae breeding in clear water irrigation ponds. however, good control of cx. quinquefasciatus was obtained using a granular formulation of b. sphaericus, vectolex-g (abg-6185), at concentrations of 10-30 kg/ha. high concentrations o ...19911895075
compatibility of bacillus thuringiensis var. israelensis and bacillus sphaericus with the fungal pathogen lagenidium giganteum (oomycetes: lagenidiales).larvae of culex quinquefasciatus were exposed to infection by lagenidium giganteum and various concentrations of b.t.i. or b. sphaericus. the resulting larval mortalities, percentages of infected dead larvae and percentages of larval body regions containing the fungus were compared. overall, the effectiveness of lagenidum giganteum against the larvae was not significantly affected by the presence of b.t.i. or b. sphaericus, and the fungal and bacterial agents were compatible. in experiments usin ...19911895077
perspectives on management of pestiferous chironomidae (diptera), an emerging global problem.in recent years, adult chironomidae emerging from some urban natural or man-made habitats have increasingly posed a variety of nuisance and economic problems, and in some situations medical problems to humans in different parts of the world. although there are an estimated 4,000 or more species of chironomid midges worldwide, less than 100 species have been reported to be pestiferous. among midge control methods, numerous laboratory and field studies have been conducted on the use of organochlor ...19911895084
aerially applied, liquid bacillus thuringiensis var. israelensis (h-14) for control of spring aedes mosquitoes in michigan.liquid b.t.i. (vectobac 12as), when mixed with water at a 1:3 ratio and applied by helicopter at a rate of 1.17 liters b.t.i. (4.68 liters mix) per ha, was 99% effective in a small (mass median diameter on dye cards: 178 microns) droplet size but ineffective (65%) in a large (553 microns) droplet against spring aedes larvae in snowmelt pools. there were about 6 times as many smaller droplets as larger ones impacting the treated pools, which probably explained the difference in effectiveness for ...19911895091
a simple technique for determining relative toxicities of bacillus thuringiensis var. israelensis formulations against larval blackflies (diptera: simuliidae).a laboratory system for assaying the potency of bacillus thuringiensis var. israelensis formulations against larval blackflies was developed. an orbital shaker was used to create a water current in 250-ml erlenmeyer flasks containing the test larvae. this system produced dose-mortality relationships with acceptable statistical parameters.19911895092
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