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1.
J Biomater Sci Polym Ed ; : 1-18, 2019 Nov 25.
Artigo em Inglês | MEDLINE | ID: mdl-31724490

RESUMO

Biological modifications of the silk fibroin (silk) material have broad applications in textiles, biomedical materials and other industrial materials. It is economical to incorporate nanoparticles to the biosynthesis of silk fibroin by adding them to silkworm larval diets. This strategy may result in the rapid stable production of modified silk. Glucose-coated silver nanoparticles (AgNPs) were used to improve the AgNPs' biocompatibility, and the AgNPs were efficiently incorporated into silk by feeding. Larvae fed with AgNPs produced silk with significantly improved antibacterial properties and altered silk secondary structures. Both positive and negative effects on the growth and synthesis of silk proteins were observed after different AgNPs doses. Larvae feeding with low concentration of 0.02% and medium 0.20% AgNPs have greater transfer efficiencies of AgNPs to silk compared with feeding high concentration of 2.00% AgNPs. In addition, the elongation and tensile strength of the produced silk fibers were also significantly increased, with greater mammalian cell compatibility. The appropriate AgNPs concentration in the diet of silkworms can promote the synthesis of silk proteins, enhance their mechanical properties, improve their antibacterial property and inhibit the presence of Gram-negative bacteria.

2.
Int J Biol Sci ; 15(10): 2170-2181, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31592097

RESUMO

Hyperproteinemia is a severe metabolic disease characterized by abnormally elevated plasma protein concentrations (PPC). However, there is currently no reliable animal model for PPC, and the pathological mechanism of hyperproteinemia thus remains unclear. In this study, we evaluated the effects of hyperproteinemia on reproductive development in an invertebrate silkworm model with a controllable PPC and no primary disease effects. High PPC inhibited the synthesis of vitellogenin and 30K protein essential for female ovarian development in the fat body of metabolic tissues, and inhibited their transport through the hemolymph to the ovary. High PPC also induced programmed cell death in testis and ovary cells, slowed the development of germ cells, and significantly reduced the reproductive coefficient. Furthermore, the intensities and mechanisms of high-PPC-induced reproductive toxicity differed between sexes in this silkworm model.

3.
Front Physiol ; 10: 537, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31130878

RESUMO

Circadian clock system disorders can lead to uncontrolled cell proliferation, but the molecular mechanism remains unknown. We used a Bombyx mori animal model of single Period gene (BmPer) expression to investigate this mechanism. A slow growing developmental cell model (Per-KD) was isolated from a B. mori ovarian cell line (BmN) by continuous knock down of BmPer expression. The effects of BmPer expression knockdown (Per-KD) on cell proliferation and apoptosis were opposite to those of m/hPer1 and m/hPer2 in mammals. The knockdown of BmPer expression led to cell cycle deceleration with shrinking of the BmN cell nucleus, and significant inhibition of nuclear DNA synthesis and cell proliferation. It also promoted autophagy via the lysosomal pathway, and accelerated apoptosis via the caspase pathway.

4.
Front Physiol ; 9: 302, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29651251

RESUMO

Hyperproteinemia, which is characterized by an abnormally elevated plasma protein concentration (PPC), is a high-mortality, metabolic complication associated with severe liver and kidney disease. It is difficult to clinically distinguish the difference between the impacts of primary diseases and hyperproteinemia on tissues and organs, and there are no available animal models of hyperproteinemia. Here, we constructed an animal model of hyperproteinemia with a controllable PPC and no primary disease effects in the silkworm Bombyx mori that has attracted interest owing to its potential use in the pathological analysis of model animals. Silkworm have an open circulatory system in which each organ is directly immersed in hemolymph. The fat body (FB) of a silkworm, as a major organ for nutrient storage and energy metabolism, can effectively reflect hyperproteinemia-induced metabolic abnormalities in damaged visceral tissues. A pathogenesis study showed that hyperproteinemia attenuated cell autophagy and apoptosis by attenuating an endocrine hormone, thereby preventing FB remodeling during metamorphosis. Meanwhile, hyperproteinemia increased oxidative stress in the FB and resulted in a dysfunction of amino acid conversion. Supplementation with exogenous 20-hydroxyecdysone effectively mitigated the hyperproteinemia-mediated inhibition of FB remodeling.

5.
Sci Rep ; 7(1): 10972, 2017 09 08.
Artigo em Inglês | MEDLINE | ID: mdl-28887546

RESUMO

Silkworms (Bombyx mori) reared on artificial diets have great potential applications in sericulture. However, the mechanisms underlying the enhancement of metabolic utilization by altering silkworm nutrition are unclear. The aim of this study was to investigate the mechanisms responsible for the poor development and low silk protein synthesis efficiency of silkworms fed artificial diets. After multi-generational selection of the ingestive behavior of silkworms to artificial diets, we obtained two strains, one of which developed well and another in which almost all its larvae starved to death on the artificial diets. Subsequently, we analyzed the metabolomics of larval hemolymph by gas chromatography/liquid chromatography-mass spectrometry, and the results showed that vitamins were in critically short supply, whereas the nitrogen metabolic end product of urea and uric acid were enriched substantially, in the hemolymph of the silkworms reared on the artificial diets. Meanwhile, amino acid metabolic disorders, as well as downregulation of carbohydrate metabolism, energy metabolism, and lipid metabolism, co-occurred. Furthermore, 10 male-dominant metabolites and 27 diet-related metabolites that differed between male and female silkworms were identified. These findings provide important insights into the regulation of silkworm metabolism and silk protein synthesis when silkworms adapt to an artificial diet.


Assuntos
Bombyx/metabolismo , Dieta , Metaboloma , Animais , Feminino , Hemolinfa/química , Masculino , Morus/química
6.
Sci Rep ; 7: 46258, 2017 04 10.
Artigo em Inglês | MEDLINE | ID: mdl-28393918

RESUMO

Abnormalities in the circadian clock system are known to affect the body's metabolic functions, though the molecular mechanisms responsible remain uncertain. In this study, we achieved continuous knockdown of B. mori Period (BmPer) gene expression in the B. mori ovary cell line (BmN), and generated a Per-KD B. mori model with developmental disorders including small individual cells and slow growth. We conducted cell metabolomics assays by gas chromatography/liquid chromatography-mass spectrometry and showed that knockdown of BmPer gene expression resulted in significant inhibition of glycometabolism. Amino acids that used glucose metabolites as a source were also down-regulated, while lipid metabolism and nucleotide metabolism were significantly up-regulated. Metabolite correlation analysis showed that pyruvate and lactate were closely related to glycometabolism, as well as to metabolites such as aspartate, alanine, and xanthine in other pathways. Further validation experiments showed that the activities of the key enzymes of glucose metabolism, hexokinase, phosphofructokinase, and citrate synthase, were significantly decreased and transcription of their encoding genes, as well as that of pyruvate kinase, were also significantly down-regulated. We concluded that inhibition of the circadian clock gene BmPer repressed glycometabolism, and may be associated with changes in cellular amino acid metabolism, and in cell growth and development.


Assuntos
Bombyx/genética , Bombyx/metabolismo , Relógios Circadianos/genética , Metabolismo Energético , Expressão Gênica , Proteínas Circadianas Period/genética , Animais , Metabolismo dos Carboidratos , Linhagem Celular , Cromatografia Líquida , Ritmo Circadiano/genética , Cromatografia Gasosa-Espectrometria de Massas , Técnicas de Silenciamento de Genes , Glucose/metabolismo , Proteínas de Insetos/genética , Redes e Vias Metabólicas , Metaboloma , Metabolômica/métodos
7.
Chemosphere ; 159: 628-637, 2016 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-27348562

RESUMO

Silicon nanoparticles (SiNPs) have attractive potential applications in biological and medical fields, and yet their impact on animals is still controversial, and there have been no reports of their effects on hematopoiesis. In this study, the effects of SiNPs on hemocytes and hematopoiesis were investigated by administering SiNPs via a vascular injection into an invertebrate model, the silkworm. Our results show that the ability of SiNPs to enter different types of circulating hemocytes and their impact on those hemocytes differed significantly. Rapid accumulation of SiNPs was observed in granulocytes, oenocytoids, and spherulocytes, which have immune functions in the circulating hemolymph, whereas SiNPs did not easily enter prohemocytes, which can differentiate into granulocytes, oenocytoids, and spherulocytes and replenish them. The SiNPs that entered the hemocytes initiated autophagy and apoptosis via the lysosomal/mitochondrial pathway. High-dose SiNPs weakly stimulated lysosomal activity in hematopoietic organs, but did not lead to a significant increase in reactive oxygen species or severe autophagy or apoptosis in the organ tissues. We suggest that the damage caused by high-dose SiNPs to hematopoiesis is self-healing, because few SiNPs entered the hematopoietic stem cells in the circulating hemolymph, so the damage to the hematopoietic tissues was limited.


Assuntos
Bombyx/fisiologia , Hematopoese/fisiologia , Hemolinfa/fisiologia , Nanopartículas/toxicidade , Silício/toxicidade , Animais , Apoptose/efeitos dos fármacos , Bombyx/efeitos dos fármacos , Hemócitos/efeitos dos fármacos , Hemolinfa/metabolismo , Mitocôndrias/metabolismo , Modelos Animais , Espécies Reativas de Oxigênio/metabolismo
8.
Sci Rep ; 6: 19802, 2016 Jan 25.
Artigo em Inglês | MEDLINE | ID: mdl-26806642

RESUMO

The use of quantum dots (QDs) in biological imaging applications and targeted drug delivery is expected to increase. However, the efficiency of QDs in drug targeting needs to be improved. Here, we show that amino acids linked to CdTe QDs significantly increased the targeted transfer efficiency and biological safety in the invertebrate model Bombyx mori. Compared with bare QDs530, the transfer efficiency of Ala- and Gly-conjugated QDs (QDs530-Ala and QDs530-Gly) in circulatory system increased by 2.6 ± 0.3 and 1.5 ± 0.3 times, and increased by 7.8 ± 0.9 and 2.9 ± 0.2 times in target tissue silk glands, respectively, after 24 h of QDs exposure. Meanwhile, the amount of conjugated QDs decreased by (68.4 ± 4.4)% and (46.7 ± 9.1)% in the non-target tissue fat body, and the speed at which they entered non-target circulating blood cells significantly decreased. The resultant QDs530-Ala revealed a better structural integrity in tissues and a longer retention time in hemolymph than that of QDs530 after exposure via the dorsal vessel. On the other hand, QDs530-Ala significantly reduced the toxicity to hemocytes, silk gland, and fat body, and reduced the amount of reactive oxygen species (ROS) in tissues.


Assuntos
Bombyx/efeitos dos fármacos , Sistemas de Liberação de Medicamentos , Pontos Quânticos/química , Aminoácidos/química , Animais , Compostos de Cádmio/química , Sobrevivência Celular/efeitos dos fármacos , Hemócitos/efeitos dos fármacos , Humanos , Invertebrados/efeitos dos fármacos , Modelos Animais , Pontos Quânticos/administração & dosagem , Espécies Reativas de Oxigênio/química , Telúrio/química
9.
Biomaterials ; 35(9): 2942-51, 2014 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-24411333

RESUMO

Quantum dots (QDs) have gained significant attention due to their superior optical properties and wide usage in biological and biomedical studies. In recent years, there has been intense concern regarding the in vivo toxicity of QDs. This study was undertaken to examine the toxicity of CdTe QDs on hematopoiesis in an invertebrate model organism, Bombyx mori. Vascular injection of sub-lethal doses of QDs in B. mori larvae caused time- and dose-dependent damage in the hematopoietic organ and hematocytes. QDs with the maximum emission wavelength of 530 nm (QDs530) were quickly observed in cystocytes and plasmacytes, and gradually bleached their green fluorescence, followed by a decrease in peripheral hematocytes. Additionally, the proportion of abnormal hematocytes increased. In marked contrast, QDs with the maximum emission wavelength of 720 nm (QDs720) were quickly surrounded by hematocytes and subsequently enriched in cystocytes like the human's leukocytes, but with weaker cytotoxicity. QDs exposure promoted the mitotic nucleus in prohemocytes and hematocytes similar to peripheral blood stem cells in humans, but aggravated apoptosis. A decrease in hematopoiesis was accompanied by shrinkage and death of hematopoietic organs via an increase in reactive oxygen species. QDs with smaller size resulted in more severe hematopoiesis toxicity.


Assuntos
Bombyx/efeitos dos fármacos , Compostos de Cádmio/toxicidade , Hematopoese/efeitos dos fármacos , Pontos Quânticos/toxicidade , Telúrio/toxicidade , Testes de Toxicidade , Animais , Apoptose/efeitos dos fármacos , Bombyx/crescimento & desenvolvimento , Contagem de Células , Núcleo Celular/efeitos dos fármacos , Núcleo Celular/metabolismo , Hemócitos/citologia , Hemócitos/efeitos dos fármacos , Hemócitos/metabolismo , Microscopia de Fluorescência , Mitose/efeitos dos fármacos , Modelos Animais , Especificidade de Órgãos/efeitos dos fármacos , Espécies Reativas de Oxigênio/metabolismo
10.
Insect Sci ; 21(1): 39-46, 2014 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-23956095

RESUMO

To explore whether glutathione regulates diapause determination and termination in the bivoltine silkworm Bombyx mori, we monitored the changes in glutathione redox cycle in the ovary of both diapause- and nondiapause-egg producers, as well as those in diapause eggs incubated at different temperatures. The activity of thioredoxin reductase (TrxR) was detected in ovaries but not in eggs, while neither ovaries nor eggs showed activity of glutathione peroxidase. A lower reduced glutathione/oxidized glutathione (GSH/GSSG) ratio was observed in the ovary of diapause-egg producers, due to weaker reduction of oxidized glutathione (GSSG) to the reduced glutathione (GSH) catalyzed by glutathione reductase (GR) and TrxR. This indicates an oxidative shift in the glutathione redox cycle during diapause determination. Compared with the 25°C-treated diapause eggs, the 5°C-treated diapause eggs showed lower GSH/GSSG ratio, a result of stronger oxidation of GSH catalyzed by thioredoxin peroxidase and weaker reduction of GSSG catalyzed by GR. Our study demonstrated the important regulatory role of glutathione in diapause determination and termination of the bivoltine silkworm.


Assuntos
Bombyx/crescimento & desenvolvimento , Bombyx/metabolismo , Glutationa/metabolismo , Animais , Bombyx/enzimologia , Feminino , Glutationa Peroxidase/metabolismo , Proteínas de Insetos/metabolismo , Metamorfose Biológica , Ovário/citologia , Ovário/enzimologia , Ovário/metabolismo , Óvulo/enzimologia , Óvulo/crescimento & desenvolvimento , Óvulo/metabolismo , Oxirredução , Tiorredoxina Dissulfeto Redutase/metabolismo
11.
Gene ; 526(2): 309-17, 2013 Sep 10.
Artigo em Inglês | MEDLINE | ID: mdl-23747351

RESUMO

The complete mitochondrial genome (mitogenome) of the Ailanthus silkmoth, Samia cynthia cynthia (Lepidoptera: Saturniidae) was determined. The circular genome is 15,345 bp long, and presents a typical gene organization and order for sequenced mitogenomes of Bombycidea species. The nucleotide composition of the genome is highly A+T biased, accounting for 79.86%. The AT skew of the genome is slightly negative, indicating the occurrence of more Ts than As, as found in other Saturniidae species. All protein-coding genes (PCGs) are initiated by ATN codons, except for COI and COII, which are tentatively designated by CGA and GTG, respectively, as observed in other insects. Four of 13 PCGs, including COI, COII, ATP6, and ND3, harbor the incomplete termination codons, T or TA. With an exception for tRNASer(AGN), all other tRNAs can form a typical clover-leaf structure of mitochondrial tRNA. The 359 bp A+T-rich region of S. c. cynthia contains non-repetitive sequences, but harbors several features common to the Bombycidea insects, including the motif ATAGA followed by a poly-T stretch of 19 bp, a microsatellite-like (AT)7 element preceded by the ATTTA motif, and a poly-A element upstream tRNAMet. The phylogenetic analyses support the morphology-based current hypothesis that Bombycidae and Saturniidae are monophyletic. Our result confirms that Saturniini and Attacini form a reciprocal monophyletic group within Saturniidae.


Assuntos
Bombyx/genética , Genoma Mitocondrial , Animais , Composição de Bases , Sequência de Bases , DNA Intergênico/química , Ordem dos Genes , Genômica , Dados de Sequência Molecular , Conformação de Ácido Nucleico , Fases de Leitura Aberta , Filogenia , RNA Ribossômico/genética , RNA de Transferência/química , RNA de Transferência/genética , Alinhamento de Sequência
12.
Anticancer Agents Med Chem ; 13(8): 1280-90, 2013 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-23796245

RESUMO

Glaucocalyxin (Gla) A-C are major ent-kauranoid diterpenoids isolated from Rabdosia japonica var. glaucocalyx, a plant used in Chinese traditional medicine as an antitumor and anti-inflammatory agent. The present investigation was carried out to observe whether cellular reduced glutathione (GSH) plays important roles in Gla -induced cytotoxicity. Among major ent-kauranoid diterpenoids isolated, Gla A and B dose-dependently decreased the growth of HL-60 cells with an IC50 of approximately 6.15 and 5.86 µM at 24 h, respectively. Both Gla A and B could induce apoptosis, G2/M-phase cycle arrest, DNA damage and the accumulation of reactive oxygen species (ROS) in HL-60 cells. Moreover, Gla A, B caused rapid decrease of the intracellular GSH content, while inhibition of cellular GSH synthesis by buthionine sulfoximine (BSO) augmented the induced cytotoxicity and apoptosis in HL-60 cells. On the other hand, the administration of GSH or GSH precursor N-acetyl-cysteine (NAC) could rescue Gla A, B-depleted cellular GSH, and abrogate the induced cytotoxicity, G2/M-phase cycle arrest, DNA damage and ROS accumulation in HL-60 cells. Furthermore, Gla A, B decreased the activity of the GSH-related enzymes including glutathione reductase (GR) and glutathione peroxidase (GPX). These data suggest that the intracellular GSH redox system plays important roles in regulating the Gla A, B-induced cytotoxicity on HL-60 cells.


Assuntos
Diterpenos de Caurano/farmacologia , Glutationa/metabolismo , Acetilcisteína/farmacologia , Apoptose/efeitos dos fármacos , Butionina Sulfoximina/farmacologia , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Glutationa Peroxidase/metabolismo , Glutationa Redutase/metabolismo , Células HL-60 , Humanos , Espécies Reativas de Oxigênio/metabolismo
13.
Chemosphere ; 93(2): 263-8, 2013 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-23719487

RESUMO

4-Nonylphenol (4-NP) a known endocrine disrupting chemical is a persistent environmental contaminant. However, the mechanism of reproductive toxicity caused by 4-NP is still largely unresolved in invertebrates. In this study, Bombyx mori larvae were constantly fed 4-NP at concentrations ranging from 0.05 to 0.4gkg(-1), reproductive toxicity and induction of vitellogenin gene (Vg) expression were investigated in this organism which is an ideal lepidopteran model insect. The results showed that gonad development was retarded and maturity was decreased in both male and female pupae, while the sex ratio was unaffected by 4-NP exposure. In the 4-NP exposed animals, the corresponding egg yolk protein, vitellin, involved in energy reserves for embryonic development in oviparous animals, was present in the testis of male pupae, and the mRNA transcript of the Vg gene was detected in the fat body, a specific organ of Vg synthesis, which is normally silent in males. In addition, expression of the Vg gene was up-regulated in the fat body of female pupae and adults, while the protein was decreased in developing eggs. Furthermore, expression of the ecdysone receptor gene (EcR) in the ovaries of pupae was down-regulated, suggested that the transport of Vg from the fat body to developing oocytes was disturbed by 4-NP due to interference in the expression of EcR related to ecdysone activity.


Assuntos
Bombyx/efeitos dos fármacos , Disruptores Endócrinos/toxicidade , Regulação da Expressão Gênica/efeitos dos fármacos , Fenóis/toxicidade , Reprodução/efeitos dos fármacos , Testes de Toxicidade , Vitelogeninas/genética , Animais , Biomarcadores/metabolismo , Bombyx/genética , Bombyx/fisiologia , Transporte Proteico/efeitos dos fármacos , Vitelogênese/efeitos dos fármacos , Vitelogeninas/metabolismo
14.
Artigo em Inglês | MEDLINE | ID: mdl-23262065

RESUMO

The dragline silk of orb-weaving spiders possesses extremely high tensile strength and elasticity. To date, full-length sequences of only two genes encoding major ampullate silk protein (MaSp) in Latrodectus hesperus have been determined. In order to further understand this gene family, we utilized in this study a variety of strategies to isolate full-length MaSp1 and MaSp2 cDNAs in the wasp spider Argiope bruennichi. A. bruennichi MaSp1 and MaSp2 are primarily composed of remarkably homogeneous ensemble repeats containing several complex motifs, and both have highly conserved C-termini and N-termini. Two novel amino acid motifs, GGF and SGR, were found in MaSp1 and MaSp2, respectively. Amino acid composition analysis of silk, luminal contents and predicted sequences indicates that MaSp1 and MaSp2 are two major components of major ampullate glands and that the ratio of MaSp1 to MaSp2 is approximately 3:2 in dragline silk. Furthermore, both the MaSp1:MaSp2 ratio and the conserved termini are closely linked with the production of high quality synthetic fibers. Our results make an important contribution to our understanding of major ampullate silk protein structure and provide a second blueprint for creating new composite silk which mimics natural spider dragline silk.


Assuntos
Estruturas Animais/metabolismo , Fibroínas/biossíntese , Fibroínas/química , Aranhas/metabolismo , Sequência de Aminoácidos , Aminoácidos/metabolismo , Animais , Sequência de Bases , Northern Blotting , DNA Complementar/genética , DNA Complementar/isolamento & purificação , Eletroforese em Gel de Poliacrilamida , Feminino , Fibroínas/genética , Interações Hidrofóbicas e Hidrofílicas , Dados de Sequência Molecular , Especificidade de Órgãos , Filogenia , Análise de Sequência de DNA , Homologia de Sequência de Aminoácidos
15.
Arch Insect Biochem Physiol ; 77(2): 72-80, 2011 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-21433065

RESUMO

For diapause eggs of the silkworm, Bombyx mori, diapause initiation is prevented with hydrochloric acid (HCl) at around 20 h post-oviposition while diapause status is terminated with chilling around 5°C. To investigate whether hydrogen peroxide (H(2)O(2)) and catalase expression are involved in diapause initiation and termination, the concentration of H(2)O(2), relatively higher levels of catalase mRNA and activity of catalase were compared between (1) 20-h-old diapause eggs and the HCl-treated diapause eggs, and (2) 10-day-old diapause eggs and the 5°C-chilled diapause eggs. Compared to diapause eggs, the HCl-treated eggs had significantly higher H(2)O(2) concentrations (up from approximately 1-3 µmol/g fresh mass to 5-8 µmol/g fresh mass), higher relative level of catalase mRNA (up from 0 to 35.2%) and higher catalase activity (up from 2.51 units/mg protein to 4.97 units/mg protein) at 96 h post-treatment. On the other hand, the 5°C chilling resulted in significant increases of H(2)O(2) concentration (up from 0.79 µmol/g fresh mass to 5.57 µmol/g fresh mass), relative level of catalase mRNA (up from 0 to 71.4%) and catalase activity (up from 0.88 units/mg protein to 3.42 units/mg protein) within 120 days. The results obtained in this work suggest that variations of H(2)O(2) and catalase expression in Bombyx eggs are involved in diapause initiation and termination.


Assuntos
Bombyx/crescimento & desenvolvimento , Catalase/genética , Catalase/metabolismo , Peróxido de Hidrogênio/metabolismo , Animais , Bombyx/metabolismo , Feminino , Regulação Enzimológica da Expressão Gênica , Ácido Clorídrico/farmacologia , Proteínas de Insetos/metabolismo , Metamorfose Biológica , Óvulo/metabolismo , RNA Mensageiro/genética
16.
Yi Chuan ; 32(9): 942-50, 2010 Sep.
Artigo em Chinês | MEDLINE | ID: mdl-20870616

RESUMO

Carotenoid-binding protein (CBP) is the only key protein that has been characterized to be involved in yellow cocoon coloration of the domesticated silkworm (Bombyx mori). Gene structure and mRNA expression profiles of cbp along with UV-Vis spectrum profiles of carotenoids in silk glands were investigated among twelve strains to disclose their relationship with cocoon color. Yellow cocoon strains of B. mori contained two or three cbp gene types, which had different mRNA products with a longer form acting functionally to code CBP protein and the smaller one without exon 2. The structures of cbp were different among the green cocoon strains with the mRNA product lacking exon 2. Only one cbp gene structure existed in white cocoon strains of B. mori, which produced the mRNA product free of exon 2. A newly identified intron 1 sequence of cbp gene in this study may have cocoon color-specificity among strains. The UV-Vis spectrum profiles of carotenoids in the yellow cocoon strains' silk glands were significantly different from those in the green cocoon strains and white cocoon strains.Together, it can be concluded that the gene structure and expression profile of cbp was closely linked to cocoon colors of B. mori.


Assuntos
Bombyx/metabolismo , Carotenoides/metabolismo , Larva/metabolismo , Seda/química , Sequência de Aminoácidos , Animais , Sequência de Bases , Bombyx/genética , Carotenoides/química , Proteínas de Transporte/química , Proteínas de Transporte/metabolismo , Cor , Genes , Proteínas de Insetos/química , Proteínas de Insetos/genética , Larva/genética , Dados de Sequência Molecular , Seda/genética
17.
Biochemistry ; 45(10): 3348-56, 2006 Mar 14.
Artigo em Inglês | MEDLINE | ID: mdl-16519529

RESUMO

Araneoid spiders use specialized abdominal glands to produce up to seven different protein-based silks/glues that have various mechanical properties. To date, the fibroin sequences encoding egg case fibers have not been fully determined. To gain further understanding of a recently reported spider silk protein gene family, several novel strategies were utilized in this study to isolate two full-length cDNAs of egg case silk proteins, cylindrical silk protein 1 (CySp1, 9.1 kb) and cylindrical silk protein 2 (CySp2, 9.8 kb), from the wasp spider, Argiope bruennichi. Northern blotting analysis demonstrated that CySp1 and CySp2 are selectively expressed in the cylindrical glands. The amino acid composition of raw egg case silk was closely consistent with the deduced amino acid composition based on the sequences of CySp1 and CySp2, which supports the assertion that CySp1 and CySp2 represent two major components of egg case silk. CySp1 and CySp2 are primarily composed of remarkable homogeneous assemble repeats that are 180 residues in length and consist of several complex subrepeats, and they contain highly homologous C-termini and markedly different N-termini. Our results suggest a possible link between CySp1 and CySp2. In addition, comparisons of stress/strain curves for dragline and egg case silk from Argiope bruennichi showed obvious differences in ultimate strength and extensibility, and similarities in toughness.


Assuntos
Proteínas de Insetos/metabolismo , Aranhas/metabolismo , Sequência de Aminoácidos , Animais , Clonagem Molecular , DNA Complementar/genética , DNA Complementar/isolamento & purificação , Feminino , Proteínas de Insetos/genética , Dados de Sequência Molecular , Óvulo/metabolismo , Alinhamento de Sequência , Análise de Sequência de Proteína , Homologia de Sequência de Aminoácidos , Seda/química , Seda/genética , Seda/metabolismo , Aranhas/genética , Relação Estrutura-Atividade
18.
Yi Chuan Xue Bao ; 32(6): 625-32, 2005 Jun.
Artigo em Chinês | MEDLINE | ID: mdl-16018190

RESUMO

On the basis of the molecular linkage map, mapmaker software QTLMapper 2.0 was used to analyze the QTLs effect of the whole cocoon weight,cocoon shell weight, ratio of cocoon shell and pupa weight of domestic silkworm. For these four cocoon quantitative traits, 7, 6, 2 and 8 effective QTLs were detected and mapped to 7, 5, 2 and 7 linkage groups, respectively. Complicated epistatic effects were found involved in the genetic variation of the whole cocoon weight and cocoon shell weight. For the whole cocoon weight, there were three pairs of QTLs with significant additive by additive interactions, in which, one pair had significant additive by dominance and dominance by dominance interactions. Whereas significant dominance were detected for three QTLs and significant additive effects one QTL had. For the cocoon shell weight, significant genetic effects, including epistatic effects were found for one pair of QTLs, significant dominance by dominance interaction for another pair of QTLs; one QTL had significant dominance and another QTL had additive by additive interaction. The ratio of cocoon shell and the pupa weight were controlled mainly by additive or dominance effects. No interaction between QTL was found for the ratio of cocoon. Most QTLs, associated with the pupa weight, had negative dominance effects. Only significant additive by additive interaction was found between one pair of QTLs. The 2nd, 3rd, 4th, 11th, 13th, 24th, 34th, 37th, and 40th linkage groups are the common chromosomal regions harboring QTLs of two or more cocoon quantitative traits. There are identical QTL or chromosomal region for the whole cocoon weight and cocoon shell weight, indicating they can be simultaneously improved by utilizing epistatic effects in breeding.


Assuntos
Bombyx/genética , Variação Genética , Locos de Características Quantitativas , Animais , Mapeamento Cromossômico , Epistasia Genética , Feminino , Masculino , Pupa/anatomia & histologia , Pupa/genética , Pupa/crescimento & desenvolvimento
19.
Yi Chuan ; 27(3): 372-6, 2005 May.
Artigo em Chinês | MEDLINE | ID: mdl-15985399

RESUMO

In this paper , the sex-effects of the cocoon quality characters in silkworm was predicted with Mixed linear model úThe fact that the probability of effect variance and predictability of random gender of whole cocoon weight, cocoon shell weight, ratio of cocoon shell and pupa weight reached a level of extreme significance showed that the gender effect of the four traits was extremely significant, which matched with the reality completely. The predictive values of gender effect of the four traits of female(male) were 0.248g (-0.247g), 2.423cg(-2.394)cg, -1.976%(1.992%) and 0.224g(-0.223g) respectively. Each trait showed single peak distribution after adjusting by sex-effects, which fitted for the request that quantitative traits should show continuously normal standard distribution if QTL analysis was taken.


Assuntos
Bombyx , Pupa , Animais , Peso Corporal , Bombyx/genética , Fenótipo , Locos de Características Quantitativas
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