• Korean Journal of Food Science and Technology
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• v.40 no.4
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• pp.470-473
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• 2008

This study examined the enzymatic digestion, average molecular weight distribution and structural changes of Bombyx mori silk gland fibroin using gel penneation chromatography and nuclear magnetic resonance methods. The pure-separation of calcium chloride-treated fibroin hydrolysates was carried out by gel filtration chromatography. Also, the effects of fibroin's enzymatic hydrolysis were investigated using an edible enzyme. The average molecular weight of three hydrolysate samples (silk gland fibroin (SF), SF-calcium chloride (SFC), and SFC-enzyme) were measured to compare their characteristics. The molecular weights of SF, SFC, and SFC-enzyme were approximately 108,000, 65,000, and 1,000 Da, respectively. Finally, we determined the structural characteristic changes of the different enzymatically digested samples by $^{13}C$ nuclear magnetic resonance methods. For the enzymatically digested fibroin, the glycine $^{13}C^{\alpha}$ resonance indicated that the amino acid was dramatically changed and/or separated out; however, this was not shown for the normal Bombyx mori silk gland fibroin.

• Journal of Sericultural and Entomological Science
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• v.38 no.2
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• pp.180-185
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• 1996

Biosynthesis tracing of the silk fibroin in Bombyx mori silkworm was examined in vivo with isotopic [1-13C] Gly. labeling by nuclear magnetic resonance method. The [1-13C] Gly. labeled silk fibroin yielded very sharp 13C NMR signal in the posterior silk gland as well as in aqueous solution and the amound of [1-13C] Gly. labeled signal in the silkworm increased gradually and rapidly to 5-th day of fifth instar. However, the decomposition or decrease of the [1-13C] Gly. labeled signal occured from 5-th to 9-th day of fifth instar unexpectedly. These findings suggest that a relative amount of ${\alpha}$-helical portion or amorphous silk II portion was formed without any further signal from 6-th day of fifth instar to pupation. Through peak separation of orientation spectrum, between the fiber axis and the molecular bond direction, N-H bond in Bombyx mori silk fiber as well as the orientation distribution around the silk fibroin axis were determined and two kinds of peaks were also obtained from this orientation spectrum.

• Journal of Life Science
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• v.10 no.1
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• pp.10-13
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• 2000

To develope the genetic source of oak wild silkworm, Antheraea yamamai, the cDNA library was constructed with poly A+ mRNA isolated from posterial silk gland of fifth instar larvae. Titer of the cDNA library was about 5.1$\times$105 pfu in total. We presumed that the titer covered almost all transcripts existed in Antherea yamamai. From cDNA library of Antheraea yamamai, fibroin heavy chain gene, which is specifically expressed from posterial silk gland of Antheraea yamamai, was screened using oligonuclotide probe specific to alanine rich motif of fibrin heavy chain gene of Antheraea pernyi. As a result, fibroin clones isolated from 5$\times$104 plaques showed the highest homolgy (95%) with that of Antherea pernyi in nucleotide of Anthereaea yamamai and Bombyx mori shows that there is no homologous sequence in the 3+ partial 채야후 region Genomic southern hybridization suggested that one copy is present. Northern hybridization showed that fibroin transcript was approximateely 9 kb in length.

• International Journal of Industrial Entomology and Biomaterials
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• v.5 no.1
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• pp.93-102
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• 2002

We have cloned and characterized the complete fibroin light chain gene from the silkworm Baekok-Jam, Bombyx mori, a recommended variety in Korea. It consists of seven exons and six introns. It consists of 14,663 nucleotides long with an open reading frame of 786 nucleotides that encodes a protein of 262 amino acid residues with a molecular mass of approximately 26,000 dalton. The amino acid alignment revealed that the Baekok-Jam fibroin light chain had 98.5 % protein sequence identity to J139 strain: differed at four amino acid positions (11, 46, 80 and 123). The Northern hybridization analysis showed that the Baekok-Jam fibroin light chain gene was specifically expressed in the posterior silk gland.

• Journal of Sericultural and Entomological Science
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• v.2
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• pp.1-31
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• 1962

The purpose of this treatise is to prove the presence of cystine in silk fiber through wide sampling throughout all the sericultural processes of Bombyx mori.; also to show that disulfide cross linkages exist in the silk fiber. The conclusions reached were as follows: 1. Crystalline cystine was obtained from silk fibroin using Folin's Method. 2. Analytical data showing the cystine content of silk fiber and its related materials were obtained using Sullvan's Method as follows: Material Percent Cystine A. Mulberry leaf protein 0.175 B. Silkworm egg 0.33 C. Silkworm Body, matured, fat extracted, without silk gland 0.41 D. Silk gland, matured 1.23 E. Silkworm feces none F. Silkworm pupa, fat extracted 0.30 G. Silkworm moth, fat extracted 0.60 H. Raw Silk 0.22 I. Fibroin 0.175 J. Sericin 0.30 3. The presence of cystine in the silkworm was substantiated the existence of 0.175 % methionine in mulberry leaves and 0.12% methionine in the silk gland. 4. Part of the sulfhydryl compounds in the silk gland is believed to transfer to serine and methionine, with the former being secreted into the liquid silk finally as silk fiber and the latter used for nutritive purposes in the growing of silk gland tissue. 5. The cystine content is variable by mulberry species, silkworm species, sex, breeding process, and other culturing environments. 6. Hybrid silkworms require more nutritive amino acids for effective growth than the original parents, and secrete less of them as silk fiber. 7. From such an observation, the amino acid composition of silk fiber is believed to be fairly flexible. Cystine if included in the amorphous part of the fiber, especially in sericin. 8. The result from enriching the silkworm diet with pure cystine or wool cystine did not result in any advantage, therefore it is believed that the natural cystine and methionine contents in the mulberry leafaregoodenoughforsilkwormnutrition. 9. The disulfide cross linkage in silk fiber was verified by using the Harris Method. Contraction took place following the treatment of the fiber with various salts and acids. Comparisons were made with wool fiber. 10. During these experiments, the fibrious structure of silk fiber and the net-globular liquid form were photographed microscopically. It is believed that the globules of liquid silk are net-formed by the inter attraction of the OH ion of the globular peptide and the H ion of water as shown by the hair cracking behavior of the film. The net-globular protein precipitation from the mulberry protein solution showed that mulberry is a proper diet for the formation of fibrous protein in the silk fiber. 11. The significance of the presence of cystine in silk fiber as emphasized in this paper should result in modification of the general conception that cystine is absent from this fiber. NOTICE: A part of this treatise was presented at the annual Korea Sericultural Society meeting held in 1961.

• Journal of Sericultural and Entomological Science
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• v.53 no.1
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• pp.44-49
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• 2015

We characterized tissue specific-expressed genes in the posterior silk gland of Bombyx mori using by the Annealing Control Primer based differential display-PCR manner. In this study, we isolated 34 differentially expressed PCR amplicons, which one of these was identified as a novel transcript named as ACP-16 (366 bp), its expression was observed only in the posterior silk glands by Northern blot analysis. To determine promoter region of the ACP-16, we isolated and analyzed a phage DNA having 1.7 kb-long genome DNA including the open reading flame and 5'- upstream untranslated region of the ACP-16 gene from a genomic DNA library. We have estimated a promoter region of the ACP-16 gene by a web promoter prediction engine, which locates -750 ~ -165 from translation initiation site (ATG, +1). ACP-16 gene is necessary to more studies about critical biological role in order to apply the silkworm's transgenic system.