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At least 631 records · Page 35Linked to original sources
PubMed1982

The nucleotide sequence of 5.8S rRNA from the posterior silk gland of the silkworm Philosamia cynthia ricini.

The nucleotide sequence of 5.8S rRNA from the Chinese silkworm Philosamia cynthia ricini has been determined by gel sequencing and mobility shift methods. The complete primary structure is (sequence in text). This is one of the largest known 5.8S rRNAs. As compared to Bombyx 5.8S rRNA, it is two nucleotides longer; two nucleotides near the 5'end and two nucleotides near the 3'end are different, and psi 61 of the Bombyx RNA sequence is an unmodified U in Philosamia RNA. The secondary structure of Philosamia 5.8S rRNA may differ from the Bombyx RNA structure by three additional base pairs at the 5'/3' ends.

Animals↗
PubMed1986

Characterization of 3'----5' exonuclease associated with DNA polymerase of silkworm nuclear polyhedrosis virus.

3'----5' Exonuclease specific for single-stranded DNA copurified with DNA polymerase of nuclear polyhedrosis virus of silkworm Bombyx mori (BmNPV Pol). BmNPV Pol has no detectable 5'----3' exonuclease activity on single-stranded or duplex DNA. Analysis of the products of 3'----5' exonucleolytic reaction showed that deoxynucleoside monophosphates were released during the hydrolysis of single-stranded DNA. The exonuclease activity cosedimented with the polymerase activity during ultracentrifugation of BmNPV Pol in glycerol gradient. The polymerase and the exonuclease activities of BmNPV Pol were inactivated by heat with nearly identical kinetics. The mode of the hydrolysis of single-stranded DNA by BmNPV Pol-associated exonuclease was strictly distributive. The enzyme dissociated from single-stranded DNA after the release of a single dNMP and then reassociated with a next polynucleotide being degradated.

Animals↗
PubMed2006

Role of the silkworm argonaute2 homolog gene in double-strand break repair of extrachromosomal DNA.

The argonaute protein family provides central components for RNA interference (RNAi) and related phenomena in a wide variety of organisms. Here, we isolated, from a Bombyx mori cell, a cDNA clone named BmAGO2, which is homologous to Drosophila ARGONAUTE2, the gene encoding a repressive factor for the recombination repair of extrachromosomal double-strand breaks (DSBs). RNAi-mediated silencing of the BmAGO2 sequence markedly increased homologous recombination (HR) repair of DSBs in episomal DNA, but had no effect on that in chromosomes. Moreover, we found that RNAi for BmAGO2 enhanced the integration of linearized DNA into a silkworm chromosome via HR. These results suggested that BmAgo2 protein plays an indispensable role in the repression of extrachromosomal DSB repair.

Amino Acid Sequence↗
PubMed1999

Two concentric protein shell structure with spikes of silkworm Bombyx mori cytoplasmic polyhedrosis virus revealed by small-angle neutron scattering using the contrast variation method.

The overall and internal structures of the silkworm Bombyx mori cytoplasmic polyhedrosis virus was investigated by small-angle neutron scattering using the contrast variation method. Data were collected in aqueous buffer solutions containing 0, 50, 75, and 100% D2O in the q range of 0.002 to 0.0774 A-1 at 5 degrees C. The radius of gyration at infinite contrast was estimated to be 336 A. The contrast matching point of the virus was determined to correspond to about 50% D2O level, evidence that the virus is composed of protein and nucleic acid. The virus was basically spherical and had a diameter of about 700 A. The main feature of its structure is the clustering of protein into two concentric shells separated by about 100 A. Most of the RNA moieties are located in the central core and between these two protein shells. However, the distance distribution function P(r) showed a minor distribution beyond a distance of r = 700 A, with a maximum particle distance of the virus of 1350 A. This is indicative of an external structure region with very low scattering density, in addition to the basic spherical structure. This external region is thought to correspond to twelve pyramidal protruding spikes shown by electron microscopic studies.

Animals↗
PubMed1988

Coupled transcription-translation of silkworm cytoplasmic polyhedrosis virus injected into oocytes of the frog, Xenopus laevis.

The cytoplasmic polyhedrosis virus of the silkworm, Bombyx mori, contains a ten-segmented, double-stranded RNA genome and five species of polypeptides, V1 (Mr 146,000), V2 (Mr 140,000), V3 (Mr 128,000)), V4 (Mr 62,000), and V5 (Mr 32,000). The virus contains an RNA-dependent RNA polymerase that transcribes the duplex genome RNA to form mRNA either in the infected animals or under appropriate conditions in vitro. We co-microinjected the virus, [alpha-32P]GTP and actinomycin D into oocytes of the frog, Xenopus laevis, and found that at least eight species of mRNA were formed in the oocytes. Virus-injected oocytes were labeled with [35S]methionine and cell extracts were treated with rabbit anti-cytoplasmic polyhedrosis virus immunoglobulin G. Analysis of the immunoprecipitates by sodium dodecyl sulfate polyacrylamide gel electrophoresis revealed that V1, V2, and V3 were produced in the virus-injected oocytes whereas V4 and V5 were not. By injecting the separated double-stranded genome segments immediately after heat-denaturation into oocytes, it was found that V1 was coded for by segment 1 with a chain length of 4.2 kilobase pairs, V2 by segment 2 or 3, whose chain lengths are both 4.0 kilobase pairs, and V3 by segment 4 with a chain length of 3.2 kilobase pairs. These results demonstrate that the Xenopus oocyte is a very useful system for the coupled transcription-translation of double-stranded RNA viruses.

Animals↗
PubMed1990

Amino acid sequence of silkworm (Bombyx mori) hemolymph antitrypsin deduced from its cDNA nucleotide sequence: confirmation of its homology with serpins.

A cDNA clone of silkworm (Bombyx mori) larval hemolymph antitrypsin (sw-AT) has been isolated from a fat body cDNA library. The cDNA has an open reading frame which codes a 392-amino acid residue polypeptide comprising a 16-residue signal peptide and a 376-residue mature sw-AT of Mr 41,805. The reactive site of sw-AT for inhibition of bovine trypsin [Sasaki, T. et al. (1987) J. Biochem. 102, 433-441] was identified as Lys343-Val344. Alignment of the sw-AT amino acid sequence with those of 11 members of the serpin superfamily of proteins clearly confirmed the homology of sw-AT with serpins. The amino acid sequence of sw-AT is 56% identical with that of the proteinase inhibitor from a lepidopteron, Manduca sexta [Kanost, M.R. et al. (1989) J. Biol. Chem. 264, 965-972], but the sequence around the reactive site shows no homology and the inhibitory specificity for proteinases is very different.

Amino Acid Sequence↗
PubMed1991

Characterization of v-sis protein expressed in silkworm larvae using the Bombyx mori nuclear polyhedrosis virus vector.

The v-sis oncogene of simian sarcoma virus encodes a protein which is homologous to the human platelet-derived growth factor B-chain. The v-sis protein undergoes a series of processing steps including dimer formation and proteolytic digestion to generate several molecular sizes of the protein. Two of these v-sis proteins were expressed alone or as polyhedrin-sis fusion proteins using the Bombyx mori nuclear polyhedrosis virus vector. The polyhedrin-sis fusion proteins contained a collagenase-sensitive site at the junction. The expression levels of the fusion proteins whose polyhedrin portions consisted of only 8 amino-terminal amino acids were 3-4 times higher than those of non-fusion proteins. One of these fusion proteins was expressed in silkworm larvae and the v-sis protein was isolated from the fusion protein by collagenolysis followed by chromatography. Because the purified v-sis protein exhibited the same molecular size on SDS-polyacrylamide gels under reducing and non-reducing conditions, it was concluded to be monomeric in structure. It possessed chemotactic activity but lacked mitogenic activity. In addition, a small amount (approximately 1%) of monomeric v-sis protein was converted in vitro to the mitogenically active v-sis protein, which could be a homo-dimer.

Amino Acid Sequence↗
PubMed1994

Amino acid sequence of an inhibitor from the silkworm (Bombyx mori) hemolymph against fungal protease.

A novel protein protease inhibitor (FPI-F) which is highly specific for fungal proteases and subtilisin was isolated from the silkworm hemolymph, and its amino acid sequence was determined by conventional methods. The inhibitor consisted of 55 amino acid residues and had a molecular weight of 6,100. The inhibitor included eight cysteine residues and relatively large amounts of acidic amino acids, but neither alanine, methionine nor tryptophan. The amino acid sequence of FPI-F was not homologous with those of other known protease inhibitors of microbe, plant or animal origin.

Amino Acid Sequence↗
PubMed1978

Chymotrypsin inhibitors from hemolymph of the silkworm, Bombyx mori.

Three new protease inhibitors were isolated and purified about 200-fold from hemolymph of silkworm larvae, Bombyx mori, using ion-exchange and affinity chromatography. Two of the three inhibitors were basic proteins (SCI-I had pI 9.4 and SCI-II had pI 9.6) and one was acidic (SCI-III had pI 4.0). The molecular weight of each inhibitor was determined to be 7,000 by the sedimentation equilibrium method. The amino acid composition of the inhibitors were similar except for the contents of Asp, Glu, Ile, Leu, and Lys. Val, His, and Trp were not present in the inhibitors and Met appeared only in SCI-III. The CD spectra of the inhibitors were all similar and indicated a low content of alpha-helical structure (10% at most). Each inhibitor could inhibit the protease and esterase activities of bovine alpha-chymotrypsin at a one-to-one molar ratio, and the dissociation constants were 3.1 X 10(-9)M for SCI-I and II and 1.3 X 10(-8)M for SCI-III. Only SCI-II showed a weak inhibitory activity against bovine trypsin. Subtilisin BPN' and papain were not inhibited by these inhibitors.

Amino Acids↗
PubMed1978

Adenylate cyclase in silkworm. Properties of the enzyme in pupal fat body.

Adenylate cyclase was assayed in a sonicated preparation of silkworm pupal fat body. The adenylate cyclase was found mostly in the particulate fraction. The activity depended upon either Mg2+ or Mn2+, and the degree of stimulation by Mn2+ was 2 times greater than that by Mg2+ compared at the saturating concentrations. In the presence of Mg2+, the enzyme was inhibited by both EGTA and high concentrations of Ca2+, showing biphasical response to Ca2+. The enzyme was stimulated several-fold by NaF. The enzyme exhibited typical Michaelis-Menten kinetics and Km values were 0.13 mM for MgATP and 0.086 mM for MnATP.

Adenylyl Cyclases↗
PubMed1981

Occurrence of thiol proteinases in the eggs of the silkworm, Bombyx mori.

In the crude extract of matured eggs of the silkworm (Bombyx mori), proteolytic activity was detected only in the acidic pH region with maximal activity at pH 3.5, and this activity was maintained throughout the development. No appreciable activity was observed in the neutral to alkaline pH region either in matured eggs or in eggs at early embryonic stages. Two molecular forms of proteinases active at pH 3.5 were obtained from matured eggs, and their molecular weights were estimated to be over 16,000 and about 68,000 respectively. An additional form of 40,000 molecular weight appeared in eggs just before hatching. They were strongly inhibited by thiol proteinase inhibitors, such as p-chloromercuribenzoate (pCMB), p-chloromercuriphenyl sulfonate (pCMPS), and N-[N-(L-3-trans-carboxyoxirane-2-carbonyl)-L-leucyl]agmatine (E-64), whereas pepstatin, diisopropylphosphorofluoridate, and EDTA were without effect, suggesting that they are thiol enzymes. They hydrolyzed casein, bovine serum albumin, egg albumin, and gamma-globulin. They could not hydrolyze alpha-N-benzoyl-DL-arginine p-nitroanilide, alpha-N-benzoyl-DL-arginine beta-naphthylamide, and several other synthetic substrates. These thiol proteinases are different from animal tissue thiol proteinases hitherto known.

Animals↗
PubMed1982

Inhibition of guanylate cyclase by guanosine 3'-phosphate and 2'-deoxyguanosine 3'-phosphate in silkworm fat body.

Guanosine 3'-phosphate and 2'-deoxyguanosine 3'-phosphate are potent inhibitors of guanylate cyclase of various tissues of the silkworm, Bombyx mori. The inhibitory activity is specific for guanine 3'-nucleotides. Thus, 2'-GMP and 5'-GMP are one order or more less potent than 3'-GMP, and the other 3'-nucleotides except 3'-IMP, i.e. 3'-AMP, 2'-deoxy-3'-AMP, 3'-CMP, and 3'-TMP, have no inhibitory activity. Guanosine and 2'-deoxyguanosine are only weakly active. The 3'-nucleotide inhibition is dependent on the concentration of Mn2+. The concentrations of 2'-deoxy-3'-GMP and 3'-GMP required for 50% inhibition are 0.12 and 0.15 mM, respectively, with 5 mM Mn2+.

Adipose Tissue↗
PubMed1984

Predominant synthesis of fibroin heavy and light chains on the membrane-bound polysomes prepared from the posterior silk gland of the silkworm, Bombyx mori.

Membrane-bound polysomes were prepared from the posterior silk gland of the silkworm, Bombyx mori, on the fourth to fifth day in the fifth larval instar. The polysomes, when supplemented with a soluble fraction from the posterior silk gland, exhibited the elongation reaction of the growing polypeptide-chains, but the initiation reaction of polypeptide synthesis was not demonstrated in this system. The predominant products synthesized on the membrane-bound polysomes were fibroin heavy chain (H-chain) and light chain (L-chain), while polypeptides of heterogeneous size classes were synthesized on the 105,000 X g-sedimentable polysomes. A substantial fraction of the fibroin L-chain synthesized was bound to the H-chain by disulfide bond. Most of the newly synthesized fibroin H- and L-chains on the membrane-bound polysomes were proved to be present within microsomal membrane vesicles because of their insensitivity to digestion with proteases in the absence of Triton X-100.

Animals↗
PubMed1985

Purification and characterization of membrane-bound alkaline proteases from midgut tissue of the silkworm, Bombyx mori.

Membrane-bound alkaline proteases from the midgut epithelia of the silkworm, Bombyx mori, were solubilized with 1% Lubrol-WX, at pH 11.2. They were purified by gel filtration on Sepharose 6B and Ultrogel AcA-202 columns and a preparative polyacrylamide gel electrophoresis. Two proteases, caseinolytic (6B3-Tc) and benzoyl-arginine-p-nitroanilide-lytic (6B3-Tb) were obtained. Both enzymes were homogeneous as judged by polyacrylamide electrophoresis. These enzymes showed high pH optima, 11.2, and pI values, above 11, and were extremely stable over a wide range of pH. The Km values for 6B3-Tb and Tc were 0.476 mM and 2.5 mg/ml respectively. Hammarsten casein and mulberry leaf protein were rapidly hydrolyzed by Tc, whereas the hydrolytic activity of Tb for Azocoll was higher than that of Tc. The protease Tb was strongly inhibited by diisopropylfluorophosphate, p-chloromercuribenzoate, benzamidine, leupeptin, and soybean trypsin inhibitor; Tc was inhibited by diisopropylfluorophosphate, tosyl phenylalanine chloromethylketone and chymostatin, but not by tosyl lysine chloromethylketone, p-chloromercuribenzoate, or iodoacetamide. The molecular weights of the proteases were estimated to be 12,800 (Tb) and 13,300 (Tc) by Sephacryl S-300 gel filtration. The amino acid analyses showed that both proteases contain a large number of acidic amino acids but a relatively small number of basic amino acids.

Amino Acids↗
PubMed1998

The functional residues and their representation by a hypothetical 3D model of silkworm eclosion hormone.

We have previously reported that an insect neuropeptide, eclosion hormone contained an alpha-helix in the N-terminal region and the helix was likely to play an important role in constructing an active globular structure. Furthermore, Met24 and Phe25 were found to be indispensable for the biological activity. On the other hand, no strict structure at the C-terminal side was found. In this paper, we predicted the secondary structure in the C-terminal side and analyzed the functional residues by a Gly-substitution technique. As a result, we speculated that the eclosion hormone contains three alpha-helices throughout the molecule which are essential for an active peptide structure. Moreover, we found four residues important for the biological activity of silkworm eclosion hormone: Phe29, Ile55, Phe58 and Leu59. In order to understand these results stereochemically, we have constructed a 3D structure using computer aided molecular modelling. The hypothetical 3D model showed that Phe25 and Phe58 interact together in a hydrophobic manner to keep a globular form. Met24, Phe29, and Ile55 are exposed to solvent to have a hydrophobic interaction with an eclosion hormone receptor. Leu59 can also play an important role by forming a functional conformation with Phe29 and Ile55.

Amino Acid Sequence↗
PubMed2005

Diagnostic and predictive value of the silkworm larvae plasma test for postoperative infection following gastrointestinal surgery.

OBJECTIVE: To determine whether the silkworm larvae plasma (SLP) test is a reliable diagnostic marker of infection in patients with infectious complications following gastrointestinal surgery. DESIGN: Prospective study. SETTING: Department of Surgery, University Hospital, Shiga University of Medical Science. PATIENTS: One hundred and twelve adult patients undergoing gastrointestinal surgery INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Thirty-nine of 112 patients developed infectious complications (infected group). Seven patients with sepsis (severely infected group) and 32 patients without sepsis (minor infected group) were observed. The operation time, blood loss, and age were significantly greater in both infected groups than in the noninfected group. The systemic inflammatory response syndrome score on postoperative day (POD) 1 and POD7 was highest in the severely infected group. The increase in C-reactive protein on POD3 and POD7 was significantly higher in both infected groups than in the noninfected group. White blood cell counts on POD7 were elevated significantly higher in the severely infected group than in the other groups. Immediately after surgery, SLP activity significantly increased compared with presurgery in all groups and was significantly higher in the minor and severely infected groups than in the noninfected group. The increased SLP activity returned to preoperative levels in the minor and noninfected groups; however, SLP activity in the severely infected groups remained high throughout the observational period. The most significant factor and time point that predicted infectious complications were the SLP test on POD1; sensitivity 66.7%, specificity 90.4%, positive and negative predictive values 78.8% and 83.5%. The area under the receiver operating characteristic curve for the SLP test was 0.813 +/- 0.046. CONCLUSIONS: The SLP test appears to be a useful marker of diagnosis and prediction of infectious complications following gastrointestinal surgery. Moreover, the SLP test may be able to evaluate not only the existence but also the severity of infection in surgical patients.

Aged↗
PubMed1999

Gene targeting in the silkworm by use of a baculovirus.

The Bombyx mori fibroin light (L)-chain gene was cloned and the green fluorescent protein (GFP) gene inserted into exon 7. The chimeric L-chain-GFP gene was used to replace the polyhedrin gene of Autographa californica nucleopolyhedrovirus (AcNPV). This recombinant virus was used to target the L-chain-GFP gene to the L-chain region of the silkworm genome. Female moths were infected with the recombinant virus and then mated with normal male moths. Genomic DNA from their progenies was screened for the desired targeting event. This analysis showed that the chimeric gene had integrated into the L-chain gene on the genome by homologous recombination and was stably transmitted through generations. The chimeric gene was expressed in the posterior silk gland, and the gene product was spun into the cocoon layer.

Animals↗
PubMed2006

Crystallization and preliminary X-ray diffraction analysis of a protease inhibitor from the haemolymph of the Indian tasar silkworm Antheraea mylitta.

A protein with inhibitory activity against fungal proteases was purified from the haemolymph of the Indian tasar silkworm Antheraea mylitta and was crystallized using the hanging-drop vapour-diffusion method. Polyethylene glycol 3350 was used as a precipitant. Crystals belonged to space group P6(3)22, with unit-cell parameters a = b = 60.6, c = 85.1 angstroms. X-ray diffraction data were collected and processed to a maximum resolution of 2.1 angstroms.

Animals↗