• BMB Reports
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• v.41 no.5
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• pp.394-399
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• 2008

Multiple sequence alignments of the Bombyx mori fibroin light chain gene (fib-L) from hybrids and from Chinese and Japanese strains demonstrated that 51.6% of the fib-L third intron is conserved. One of these conserved segments, 41 bp long, contains the sequence CGTTATTATACATATT, which is duplicated in the B. mori Nd-$s^D$ mutant. In the present work, electrophoretic mobility assays and computational analyses revealed a major peak of intrinsic bent DNA within the segment that undergoes breakage in the previously-described Nd-$s^D$ mutation. This result suggested that this intrinsically-curved region might mediate DNA cleavage and enhance recombination events in the third intron of the Bombyx mori fib-L gene.

• Journal of Sericultural and Entomological Science
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• v.36 no.2
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• pp.138-146
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• 1994

The structural characteristics of Antheraea yamamai and Antheraea pernyi silk were investigated by using x-ray diffraction method, IR spectroscopy and polarizing microscopy. The amino acid composition, fiber density, thermal decomposition temperature and glass transition temperature were also measured for relating these physical properties to the structure in comparison with those of Bombyx mori silk fiber. There was no significant structural difference between A. yamamai and A. pernyi silk fiber on an examination of x-ray diffraction curve and IR spectrum. Both of these wild silk fibers showed double diffraction peaks at the Bragg angle 2Θ16.7˚ and 20.5˚by x-ray diffraction analysis as well as IR absorption peaks for the bending vibration of specific groups related to ala-ala amino acid sequence. On the other hand, the x-ray diffraction curve and IR spectrum of Bombyx mori silk fiber are different from those of wild silk fibers, indicating different crystal structure as well as amino acid sequences. It showed under the polarizing microscope examination that the birefringence and optical orientation factor of wild silk fibers are much lower than those of B. mori silk. Also, the surface of degummed wild silk fibers was characterized by the longitudinal stripes of microfibrils in the direction of fiber axies. The amino acid composition, which is strongly related to the fine structure and properties, was not significantly different between these two wild silk fibers. However, the alanine content was somewhat less and polar amino acid content more for A. yamamai. As a result of fiber density measurement, the specific gravities of B. mori, A. pernyi and A. yamamai were 1.355~1.356, 1.308~1.311, 1.265~1.301g/㎤ in the order, respectively. The calculated crystallinity(%) was 64% for B. mori and 51~52% for wild silk fibers, which showed same trend by IR method in spite of somewhat higher value. The thermal decomposition behaviour was examined by DSC and TGA, showing that the degradation temperature was in the order of B mori, A. prernyi and A. yamamai at around 350$^{\circ}C$. It was also observed by TGA that the decomposition seems to proceed step by step according to their specific regions in the fiber structure, resulting the difference in their thermal stabilities. The glass transition temperature was turned out to be 220$^{\circ}C$ for B. mori, 240$^{\circ}C$ A. yamamai and 255$^{\circ}C$ A. pernyi by the dynamic mechanical analysis. It is expected that the chemical properties are affected by the dynamic mechanical behavior in accordance with their structural characters.

• International Journal of Industrial Entomology and Biomaterials
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• v.37 no.1
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• pp.21-28
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• 2018

CRISPR/Cas9 gene editing system is an efficient method to mutation in a sequence specific manner. Here we report the direct transfection of the Cas9 nuclease and gene specific guide RNA can be used in BM-N cell line derived from Bombyx mori ovarian tissue to enfeeble function of endogenous gene in vitro. We have used gene editing system to negative regulation components of major signaling cascade, the Toll pathway, which controls B. mori resistance to microbe infections, such as fungi and gram positive bacteria. We demonstrate that the $I{\kappa}B-like$ protein Cactus may controls the activation of transcription factors such as Rel A and Rel B. The direct transfection of Cas9 nuclease and Cactus-specific guide-RNA complex may be used in BM-N cells to disrupt the function of endogenous genes in vitro. A mutation frequency of 30-40% was observed in the transfected cells, and various mutations caused the target region. Moreover, RT-PCR analysis revealed that Cactus gene was down regulated after these mutations. More importantly, mutation of BmCactus stimulated expression of lysozyme, moricin, and lebocin genes. These results suggest that the CRISPR/Cas9 systems are expected to efficiently induce site-specific mutations and it was possible to produce antimicrobial peptide through the gene editing.

• International Journal of Industrial Entomology and Biomaterials
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• v.7 no.2
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• pp.127-131
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• 2003

Protein disulfide isomerase (PDI) found in the endoplasmic reticulum (ER) catalyzes disulfide bond exchange and assists in protein folding of newly synthesized proteins. PDI also functions as a molecular chaperone and has been found to be associated with proteins in the ER. In addition, PDI functions as a subunit of two more complex enzyme systems: the prolyl-4-hydroxylase and the triacylglycerol transfer proteins. A cDNA that encodes protein disulfide isomerase was previously isolated from Bombyx mori (bPDI), in which open reading frame of 494 amino acids contained two PDI-typical thioredoxin active site of WCGHCK and an ER retention signal of the KDEL motif at its C-terminal, and we report its functional characterization here. This putative bPDI cDNA is expressed in insect Sf9 cells as a recombinant proteins using baculovirus expression vector system. The bPDI recombinant proteins are successfully recognized by antirat PDI antibody, and shown to be biologically active in vitro by mediating the oxidative refolding of reduced and scrambled RNase. This suggests that bPDI may play an important role in protein folding mechanism of insects.

• Journal of Physiology & Pathology in Korean Medicine
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• v.24 no.3
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• pp.463-469
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• 2010

This research is performed to obtain positive evidences of Mori cortex, a kind of oriental medicinal herbs, in the cellular levels. The extracts of M. cortex have shown anti-inflammatory effects against cutaneous inflammation and clinical effects on pulmonary asthma and congestion in oriental medicine. Thus BV2 cells were chosen because microglia are considered as the main immunocompetent cells in the central nervous system. Lipopolysaccharide (LPS)-induced microglial activation of cultured BV2 cells and subsequent release of nitric oxide (NO) and Prostaglandin E2 (PGE2) were effectively suppressed by methylene chloride extract of Morus alba L. (MEMA). From the inflammation-mediated mRNA and protein analyses, we showed that inducible NO synthase (iNOS), cyclooxygenase-2 (COX-2), interleukin-$1{\beta}$ (IL-$1{\beta}$) and tumor necrosis alpha (TNF-${\alpha}$) induced by LPS were markedly decreased by MEMA treatment. From the observation of nuclear factor-kB (NF-${\kappa}B$) which is controlling and mediating inflammation through COX-2 and iNOS, there showed that p65, a subunit of NF-${\kappa}B$, was increased in nuclear and $I{\kappa}B$, a competitor of NF-${\kappa}B$, was recovered in cytosol after MEMA treatment. These are corresponding with results of iNOS, COX-2, IL-$1{\kappa}$ and TNF-${\alpha}$, and confirm some suppressive effect against transcriptional activation of NF-${\kappa}B$. In conclusion, the anti-inflammatory action of M. cortex against BV2 microglia cells is expected to protect nerve tissues through suppression of neuronal inflammation in various neurodegenerative diseases.

• International Journal of Industrial Entomology and Biomaterials
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• v.23 no.2
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• pp.223-229
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• 2011

Cyclophilins are originally identified as cytosolic binding protein of the immunosuppressive drug cyclosporine A. They have an activity of peptidyl prolyl cis/trans-isomerases (PPIase), which may play important roles in protein folding, trafficking, assembly and cell signaling. In this study, we report the cloning and characterization of a Bombyx mori cyclophilin A (bCypA) cDNA. The full-length cDNA of bCypA consist of 947 nucleotides with a polyadenylation signal sequence AATAAA and contain an open reading frame of 498 nucleotides encoding a polypeptide of 166 amino acids. The deduced amino acid sequence of bCypA shares a central peptidyl prolyl cis/trans-isomerase and a cyclosporin-A-binding domain with other cyclophilin sequences. Relative quantification real-time (RT) PCR analysis shows that mRNA transcripts of bCypA are detected in all the investigated tissues and highest expression level in the skin of 3-day-old 5 instar larva. Also, bCypA had PPIase activity on the proline-containing peptides. Accordingly, we suggest that bCypA is a new member of the cyclophilin A (CyPA) family and will be useful for quality control of bioactivity recombinant proteins with proline-containing peptides.

• Journal of Sericultural and Entomological Science
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• v.33 no.2
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• pp.75-81
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• 1991

The study has been carried out to acquire some basic informations about Bacillus thuringiensis for developing the microbial pesticide. Pathogenicity tests on three of B.thuringiensis var. aizawai, kurstaki, and dendrolimus were determined in two species of insects, H. cunea and B. moris. The pathogencity in varieties of B. thuringiensis against H. cunea and B. mori was depended on instar age of tested larvae. Bacillus thuringiensis var. dendrolimus, kurstaki, aizawai are arranged in order of pathogenicity against H. cunea and B. mori. In result Bacillus thuringiensis var. kurstaki was shown the most stable toxicity with respect to each instar of tested larvae.

• Korean journal of applied entomology
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• v.34 no.4
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• pp.344-349
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• 1995

To isolate naturally occurring novel Bacillus thuringiensis strains for useful pest con시 agents, A total of 64 isolates of B. thuringiensis producing spore and crystal were obtained from 583 soil samples in Korea. Toxicity of isolates was assayed against Lepidoptera (Bombyx ori), Diptera (Culex pipens) and Coleoptera (Sitophilus oryzae), respectively. The results showed that B. thuringiensis isolates toxic Lepidoptera are 42.2% , both Lepidoptera and Diptera are 31.3% and Diptera are 20.3%, demonstrating that spectrum of insecticidal activity of B. thuringiensis isolates is various. But no B. thuringiensis isolates have toxicity against Coleoptera.

• Korean journal of applied entomology
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• v.42 no.4
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• pp.353-360
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• 2003

Immunodepression can be required for entomopathogenic bacteria to induce their potent pathogenicities to the target insects. Here, we raise a hypothesis that the capacity of a pathogenic bacterium to induce the target insect immunodepression has positive relationship with the degree of pathogenicity. X. nematophilus had 1,200 times as potent as another entomopathogenic bacterium, Staphylococcus gallinarum against the fifth instar larvae of silkworm, Bombyx mori, when they were Injected into the hemocoel. Although both bacteria had significant cytotokic effect on the hemocytes of B. mori, X. nematophilus gave faster and greater cytotoxicity than did S. gallinarum. In cellular immune reactions, B. mori could form 20 hemocyte nodules against the bacterial injection with 5${\times}$10$\^$5/ cells. The number of the hemocyte nodules was significantly depressed when live X. nematophilus was inject-ed, but not in S. gallinarum. Activation of prophenoloxidase (proPO) was depressed in the bacterial injection. The depression of PO activation was significantly greater in X. nematophilus infection than in S. gallinarum injection. Lysozyme activity was induced by the injection of S. gallinarum at 4 h after the treatment, but not induced in X. nematophilus at all the time. These results showed that X. nemato-philus induced greater immunodepression against B. mori and resulted in higher pathogenicity than did S. gallinarum. Therefore, this study suggests that the immunodepression induced by entomopathogenic bacteria has positive relationship with their pathogenicity.

• International Journal of Industrial Entomology and Biomaterials
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• v.3 no.1
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• pp.83-88
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• 2001

The effect of topical application with 200,400 and 600 $\mu$g/ml 2,4-dichlorophenoxyacetic acid (2,4-D) on the fifth larval stadium of the silkworm, B. mori, was analyzed. Larvae treated during fifth larval stadium enhanced larval, cocoon and adult parameters. The larval period was significantly decreased with increase in silk gland weight at 400 and 600 $\mu$g/ml, male cocoon weight and shel1 weight in all the treated groups and filament length and weight at 200 $\mu$g/ml treated group. Length of the ovariole, eggs per ovariole and hatching percentage increased significantly in all the treated groups when compared with those of the carrier control. This suggests that the plant growth regulator 2,4-D in addition to affecting silk production also affect reproductive performance.