• Title/Summary/Keyword: Silk gland

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Manufacture and Characterization of Silkworm Gland Hydrolysate (누에 실샘 가수분해물의 제조 및 특성 규명)

  • Hwang, Jung Wook;Lee, Heui Sam;Kim, Hojin;Kim, Kyu-Oh;Choi, Yong-Soo
    • Journal of Sericultural and Entomological Science
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    • v.50 no.2
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    • pp.76-81
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    • 2012
  • Silk protein has been explored to be used for biomedical applications for several decades. However, it has not been used in this field cause to their irreversible crystallization after dissolving in water. The existing methods of silk protein hydrolysis using silkworm cocoon were used with harmful solvents and through a very complicated process. Therefore, we have developed novel methods for the production of water-soluble hydrolysate using silkworm gland. We manufactured two types of silkworm gland-derived hydrolysate (water-soluble SGH, SSGH; total SGH, TSGH) and compared the characteristics with commercial cocoon-derived sericin hydrolysate (CSH). The molecular weight of SGH ranged from 7 to 50 kDa (SSGH) and 5 to 15 kDa (TSGH) within glycine, alanine, and aspartic acid as a main amino acid composition. In contrast, CSH ranged from 15 to 50 kDa within serine and aspartic acid. The results of FTIR implied that SGH was more soluble form than CSH, as shown by the decrease in the ${\beta}$-sheet structure at $1630cm^{-1}$ on amide I peak. In comparison with 10% fetal bovine serum, 0.1% (1 mg/ml) SSGH had equivalent effect on the proliferation of human dermal fibroblasts and mesenchymal stem cells. All results of the SSGH made by novel manufacturing process indicate the SSGH is more preferable as a culture medium supplement than cocoon-derived sericin.

Identification of Highly Transcribed Genes in Japanese Oak Silkworm, Antheraea yamamai, Using PCR-Based cDNA Library

  • Lee, Jin-Sung;Kim, Ki-Hwan;Goo, Tae-Won;Yun, Eun-Young;Kang, Seok-Woo;Suh, Dongs-Sang;Hwang, Jae-Sam
    • International Journal of Industrial Entomology and Biomaterials
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    • v.1 no.2
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    • pp.171-175
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    • 2000
  • Determined sequences of 384 randomly selected clones in a PCR-based cDNA library of Antheraea yamamai could identify expressed sequence tags (ESTs) of highly expressed gene. One EST (fibroin) appeared 15 times, one EST (40S ribosomal protein S18) twelve times, one EST (ribosomal protein S24a) eleven times, ten times (ribosomal protein S8), nine times (60S ribosomal protein L10A), seven times (60S ribosomal protein S15A, S17, S17 and seroin), six times (ribosomal protein S8), five times (ribosomal protein S24, mariner transposase and P8 protein), four times (serpin 2), three times (heat shock protein 70 and poly A binding protein), and the remaining 6 ESTs twice (amylase, KIAA1006, elongation factor-1, transposon mag, translation initiation factor 4C, QM protein, transposase). Therefore, the 94 EST make it possible to identify 24 redundant clones that are candidates for highly expressed genes in posterior silk gland of this insect. The 24 redundant EST clones were identified in GenBank, but none of them was related to A. yamamai, suggesting that there are many unidentified genes which are highly expressed in the A. yamamai genome.

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Cloning and Spatiotemporal Expression Analysis of Bombyx mori elav, an Embryonic Lethal Abnormal Visual Gene

  • Wang, Geng-Xian;Liu, Ying;Sim, Yang-Hu;Zhang, Sheng-Xiang;Xu, Shi-Qing
    • International Journal of Industrial Entomology and Biomaterials
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    • v.18 no.2
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    • pp.113-120
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    • 2009
  • Embryonic lethal abnormal visual (elav) is a lethal gene in Drosophila inducing the abnormal development and function of nervous system. We cloned a Bm-elav gene by bioinformatics and biological experiment, based on sequence of ELAV protein and dbEST of Bombyx mori. The full-length of Bm-elav cDNA is 1498 bp, contains a 906 bp open read frame (ORF) encoding a precursor of 301 amino acid residues with a calculated molecular weight of 34 kDa and pI of 8.99. Bm-ELAV protein precursor contains three RNA recognition motifs (RRM) in $24{\sim}91$, $110{\sim}177$ and $222{\sim}295$ bit amino acid residues respectively, and belongs to RNA-binding protein family. Bm-ELAV shared varying positives, ranging from 56% to 60% (Identities from 41% to 45%), with RRM from other species of Xenopus tropicalis, Apis mellifera, Tribolium castaneum, Branchiostoma belcheri and Drosophila. Gene localization indicated that Bm-elav is a single-copy gene, gene mapping within 12-chromosome from 7916.68 knt to 7918.16 knt region of nscaf2993. Spatiotemporal expressions pattern analysis revealed that Bm-elav expressed higher in most tested tissues and developmental stages in whole generation, such as silk gland, fat body, midgut, hemopoietic organ and ovary, but almost no expression in terminated diapause eggs. This suggested that the expression of Bm-elav in early developmental embryonic stages might induce abnormal development like in Drosophila. Cloning of the Bm-elav gene enables us to test its potential role in controlling pests by transferring the gene into field lepidopteran insects in the future.

Single-dose oral toxicity study of genetically modified silkworm expressing EGFP protein in ICR mouse

  • Jang, Kyung-Min;Kim, Sung-Gun;Park, Ji-Young;Choi, Won-Ho;Lee, Jae-Woo;Jegal, Hyeon-Young;Kweon, Soon-Jong;Choi, Kwang-Ho;Park, Jung-Ho
    • Korean Journal of Agricultural Science
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    • v.43 no.1
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    • pp.109-115
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    • 2016
  • Silk has had a reputation as a luxurious and sensuous fabric but it is not popular due to the expensive price and poor durability. To develop the silk materials that apply the various industries, the artificially synthesized gene can be introduced into the silkworm and expressed in the silk gland. Transgenic silkworms for the mass production of green fluorescent silks are generated using a fibroin H-chain expression system. For commercial use, safety assessment of the transgenic silkworms is essential. The purpose of this study was to examine the potential acute oral toxicity of EGFP protein expressed in genetically modified (GM) fluorescence silkworm and to obtain the approximative lethal dose in the male and female at 6-weeks ICR mice. EGFP protein was fed at a dose of 2,000 mg/kg body weight in five male or five female mice. Mortalities, clinical findings and body weight changes were monitored for 1, 3, 7, 14 days after dosing. At the end of 14 day observation period, all mice were sacrificed, and the postmortem necropsy were performed. The test group was not observed death case. Also the effect was not admitted by test substance administration in common symptoms, the body weight and postmortem. The results of single-dose oral toxicity test showed that approximative lethal dose of EGFP protein expressed in fluorescence silkworm was considered to exceed the 2,000 mg/kg body weight in both sexes.

Ultrastructure of the Ampullate Gland in the Orb Web Spider, Nephila clavata L. Koch I. Excretory Duct of the Large Ampullate Gland (무당거미(Nephila clavata L. Koch) 병상선(甁狀腺)의 미세구조(微細構造) I. 대병상선(大甁狀腺)의 분비관(分泌管))

  • Moon, Myung-Jin;Kim, Chang-Shik;Kim, Woo-Kap
    • Applied Microscopy
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    • v.18 no.2
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    • pp.77-90
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    • 1988
  • The ultrastructure of the excretory duct of the large ampullate gland in the orb web spider, Nephila clavain L. Koch are studied with light and electron microscopes. The excretory ducts of the large ampullate glands connected with the large spinning tubes(spigots) on the anterior spinnerets are basically composed of three superposed types of the layers which are inner cuticles, monolayered epithelial cells and peripheral connective cells. According to the morphological characteristics of the cuticles and internal textures of the epithelial cells, the long excretory ducts are subdivided into three(distal, middle and proximal) portions. Especially, at the distal portion of the ducts near the spinning tubes, the electron lucent subcuticles which had the functions of water removal and orientation of silk fibers are well distributed, whereas at the middle and proximal portions these layers disappeared and instead of these, endocuticles are developed. The endocuticle contains two types of bands, which are electron dense and electron lucent. And along the length of the cuticular stem in the excretory duct, these two alternating bands are twisted spirally. In the cytoplasm of the columnar epithelial cells of the distal portion, rough endoplasmic reticula and Golgj complexes, related to the production of the cuticular materials are well developed. Between the adjacent epithelial cells, specialized septate junctions and desmosomes are formed along the plasma membranes. At the proximal portion of the duct, densely accumulated secretory materials appeared, and these are released to the inner canal by the apocrine secretion.

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Fine Structure of the Silk Spigots in the Spider Dolomedes sulfureus (Araneae: Pisauridae) (닷거미과의 황닷거미(Dolomedes sulfureus)의 실크 분비장치의 미세구조)

  • Moon, Myung-Jin
    • Applied Microscopy
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    • v.38 no.2
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    • pp.89-96
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    • 2008
  • Dolomedes spiders of the family Pisauridae are one of free wandering spiders with semi-aquatic habitation. They do not build web for prey-hunting but build a nursery web for spiderlings. This paper describes the fine structure of the silkspinning spigots of the fishing spider Dolomedes sulfureus revealed by the field emission scanning electron microscope (FESEM). The fishing spider Dolomedes sulfureus possesses only three types of silk glands which connected through the typical spinning tubes on the spinnerets. The silk spigots of this spider were identified as three groups: ampullates, pyriforms and aciniforms. Two pairs of major ampullate glands send secretory ductules to the anterior spinnerets, and another two pairs (or $1{\sim}2$ pairs in males) of minor ampullate glands supply the middle spinnerets. In addition, the pyriform glands feed silk into the anterior spinnerets ($62{\sim}68$ pairs in females and $45{\sim}50$ pairs in males), and the aciniforms send ductules to the middle ($33{\sim}40$ pairs in females and $18{\sim}25$ pairs in males) and the posterior spinnerets ($42{\sim}50$ pairs in females and $24{\sim}28$ pairs in males). Among these, the ampullate one is the most predominate gland in both sexes.

The Effect of Mineral Salts, Magnesium Sulphate and Potassium Nitrate on the Economic Parameters of Silkworm Bombyx mori L.

  • Goudar, K.S.;Kaliwal, B.B.
    • International Journal of Industrial Entomology and Biomaterials
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    • v.3 no.1
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    • pp.69-73
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    • 2001
  • The effect of topical application 100,300 and 500 $\mu$g/ ml with Mineral Salts of magnesium sulphate and potassium nitrate on economic parameters was ana- lyzed following treatment of last larval stadium. The treated larvae showed significantly decreased larval weight and silk gland weight in all the treated groups along with other decreased Iarval, cocoon and adult parameters. The female cocoon weight was significantly decreased in all the treated groups with decreased female cocoon shell weight and male cocoon weight at 300 and 500$\mu$g/ml respectively. The length and weight of filament was significantly decreased at 300 and 500 $\mu$g/ml respectively and denier at 500 $\mu$g/ ml. The fecundity decreased significantly in 300 and 500 $\mu$g/ml treated groups when compared with the corresponding parameters of the carrier control.

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