• Title/Summary/Keyword: Bombyx

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Synchronized Expression of Two Bombyx mori Caspase Family Genes, ice-2 and ice-5 in Cells Induced by Ultraviolet Irradiation

  • Wang, Wenbing;Sun, Ying;Song, Lina;Wu, Yan;Wu, Huiling
    • International Journal of Industrial Entomology and Biomaterials
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    • v.17 no.1
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    • pp.121-124
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    • 2008
  • The caspase family proteins play an important role in programmed cell death (apoptosis). To date, the expression profiles of the caspase family genes in Bombyx mori (Bm) are poorly known. In this study, we examined the expression profiles of two novel Bm caspase family genes (ice-2 and ice-5), the potential change of the mitochondrial membrane and the morphology in Bm cells after stimulation of ultraviolet (UV) irradiation. The results showed the potential change of the mitochondrial membrane occurred at 5 hours after UV irradiation treatment. Analysis of fluorescent quantitative RT-PCR demonstrated that both the ice-2 and ice-5 might be involved in UV induced apoptosis in Bm cells. Notably, after UV irradiating, expression pattern of ice-2 and ice-5 were remarkably different. The ice-2 gene was highly expressed at two time points, 0.5 and 5 hours after UV stimulating, while the expression level of ice-5 only peaked at 5 hours after UV stimulating. It indicated that apoptosis induced by UV irradiation was involved in the mitochondrial pathway and the two isoforms of Bm ice may act but play different role during the apoptosis of Bm cells.

Enzymatic activity of Endoplasmic Reticulum Oxidoreductin 1 from Bombyx mori

  • Park, Kwanho;Yun, Eun-Young;Goo, Tae-Won
    • International Journal of Industrial Entomology and Biomaterials
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    • v.37 no.1
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    • pp.15-20
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    • 2018
  • Most proteins produced in the endoplasmic reticulum (ER) of eukaryotic cells fold via disulfide formation (oxidative folding). Oxidative folding is catalyzed by protein disulfide isomerase (PDI) and PDI-related ER protein thiol disulfide oxidoreductases (ER oxidoreductases). In yeast and mammals, ER oxidoreductin-1s (ERO1s) supply oxidizing equivalent to the active centers of PDI. We previously identified and characterized the ERO1 of Bombyx mori (bERO1) as a thioredoxin-like protein that shares primary sequence homology with other ERO1s. Here we compare the reactivation of inactivated rRNase and sRNase by bERO1, and show that bERO1 and bPDI cooperatively refold denatured RNase A. This is the first result suggesting that bERO1 plays an essential role in ER quality control through the combined activities of bERO1 and bPDI as a catalyst of protein folding in the ER and sustaining cellular redox homeostasis.

Expression and In Vitro Activity of Recombinant Canstatin in Stably Transformed Bombyx mori Cells

  • Lee, Ji-Hye;Lee, Jong-Min;Jeon, Hwang-Bo;Shon, Bong-Hee;Yang, Jai-Myung;Chung, In-Sik
    • Journal of Microbiology and Biotechnology
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    • v.19 no.7
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    • pp.685-689
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    • 2009
  • We describe the expression of recombinant canstatin from stably transformed Bombyx mori BmS (BmS) cells. Recombinant canstatin was secreted into a culture medium with a molecular mass of approximately 29 kDa. Densitometric scanning showed that the secreted canstatin accounted for approximately 91% of the total canstatin production. Recombinant canstatin was also purified to homogeneity using a simple one-step Ni-NTA affinity fractionation. The identity of the purified protein was confirmed as human canstatin by nano-LC-MS/MS analysis. Purified recombinant canstatin inhibited human endothelial cell proliferation in a dose-dependent manner. The concentration at half-maximum inhibition ($ED_{50}$) for recombinant canstatin expressed in stably transformed BmS cells was approximately 0.64 ${\mu}g/ml$. A maximum production level of 11 mg/l recombinant canstatin was obtained in a T-flask culture of BmS cells after 6 days of incubation.

Genomic Organization of Heat Shock Protein Genes of Silkworm Bombyx mori

  • Velu, Dhanikachalam;Ponnuvel, Kangayam M.;Qadri, Sayed M. Hussaini
    • International Journal of Industrial Entomology and Biomaterials
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    • v.15 no.2
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    • pp.123-130
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    • 2007
  • The Hsp 20.8 and Hsp 90 cDNA sequence retrieved from NCBI database and consists of 764 bp and 2582 bp lengths respectively. The corresponding cDNA homologus sequences were BLAST searched in Bombyx mori genomic DNA database and two genomic contigs viz., BAAB01120347 and AADK01011786 showed maximum homology. In B. mori Hsp 20.8 and Hsp 90 is encoded by single gene without intron. Specific primers were used to amplify the Hsp 20.8 gene and Hsp 90 variable region from genomic DNA by using the PCR. Obtained products were 216 bp in Hsp 20.8 and 437 bp in Hsp 90. There was no variation found in the six silkworm races PCR products size of contrasting response to thermal tolerance. The comparison of the sequenced nucleotide variations through multiple sequence alignment analysis of Hsp 90 variable region products of three races not showed any differences respect to their thermotolerance and formed the clusters among the voltinism. The comparison of aminoacid sequences of B. mori Hsps with dipteran and other insect taxa revealed high percentage of identity growing with phylogenetic relatedness of species. The conserved domains of B. mori Hsps predicted, in which the Hsp 20.8 possesses ${\alpha}-crystallin$ domain and Hsp 90 holds HATPase and Hsp 90 domains.

Expression of Bombyx mori Nucleopolyhedrovirus ORF4 under the Control of BaculoviruS Ie1 Promoter by a Novel Bac-to-Bac/BmNPV Baculovirus Expression System

  • Su, Wujie;Wu, Yan;Wu, Huiling;Wang, Wenbing
    • International Journal of Industrial Entomology and Biomaterials
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    • v.15 no.2
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    • pp.131-135
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    • 2007
  • Open reading frame 4 of Bombyx mori nucleopolyhedrovirus (BmNPV), designated as Bm4, is a gene whose function is completely unknown. With the recently developed BmNPV bacmid and a modified pFastBac1 whose polyhedrin promoter was replaced with ie1 promoter, a recombinant bacmid expressing Bm4-EGFP fusion protein under the control of ie1 promoter in BmN cells was successfully constructed. The result not only showed that the polyhedrin promoter can be replaced efficiently with other promoters to direct the expression of foreign gene in BmN cells by using Bac-to-Bac/BmNPV baculovirus expression system but also laid the foundation for rescue experiment of Bm4 deletion mutant due to the ability of ie1 promoter to direct gene expression throughout the infection cycle.

Influence of Temperature and Relative Humidity in Infection of Nosema bombycis (Microsporidia: Nosematidae) and Cross-infection of N. mylitta on Growth and Development of Mulberry Silkworm, Bombyx mori

  • Chakrabarti, Satadal;Manna, Buddhadeb
    • International Journal of Industrial Entomology and Biomaterials
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    • v.17 no.2
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    • pp.173-180
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    • 2008
  • The influence of temperature and relative humidity in infection and cross-infection of Nosema bombycis and N. mylitta respectively in mulberry silkworm, Bombyx mori L. on larval mortality, multiplication of pathogens, larval weight and growth rate in three different seasons were studied. Seasons were selected in such condition, when very less fluctuations between minimum and maximum temperature and minimum and maximum relative humidity ($25{\sim}28^{\circ}C$ and $65{\sim}72%$ R.H) was observed i.e., season-1. Fluctuations between minimum and maximum temperature were less ($28.05{\sim}34.50^{\circ}C$) but R.H % was more ($55{\sim}81%$) in season-2. Fluctuations between minimum and maximum temperature and R.H % were more ($20.00{\sim}40.5^{\circ}C$ and $64.00{\sim}90.00%$) in season-3. Growth rate of microsporidian-infected silkworm is directly related to the prevailing temperature and relative humidity in silkworm. Silkworm can tolerate slight variation of temperature but slight variation of relative humidity disfavours the development of silkworm and favours the multiplication of pathogens.

Accumulation of Dietary Conjugated Linoleic Acid (CLA) in Silkworm, Bombyx mori

  • Park, Chung-Gyoo;Kim, Seck-Jong;Ha, Nam-Gue;Park, Bong-Il;Ha, Yeong-Lae
    • Preventive Nutrition and Food Science
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    • v.6 no.1
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    • pp.19-22
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    • 2001
  • Conjugated linoleic acid (CLA) exhibits potent anti-carcinogenic and other biological activities in several animal models. We report here that dietary CLA, chemically synthesized from corn oil, accumulates I silkworm, Bombyx mori, which is used as therapeutic agent for diabetes in Korea and Japan. Mulberry leaves treated with 0.1 or 10% CLA in ethanol were supplied to silkworms from the end of 3rd instar to the 3rd day of the 5th instar. Fresh mulberry leaves or leaves treated with 10% corn oil in ethanol were fed as a check treatment. the amount of total lipids in the larval body ranged from 17.4 to 19.1mg/g of body tissue, which was not significantly affected by the source of the diets. No CLA was found in the control silkworms. But the level of CLA significantly increased to 83.5 mg/g of fat, when fed with mulberry leaves treated with 10% CLA. Only trace amounts of CLA were detected in the larvae reared with check leaves and 0.1% CLA-treated leaves. Mulberry leaves treatd with corn oil or CLA were not palatable to the larvae, resulting in a reduction of larval weight. These results suggest that silkworms containing CLA in body lipids could be producted by dietary CLA.

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Distribution and Characterization of Bacillus thuringiensis isolated fro soils in Korea (한국 토양에서 분리된 Bacillus thuringiensis의 분포와 특성조사)

  • 김호산;박현우;이대원;유용만;김정일;강석권
    • 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.

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Characterization of Bacillus thuringiensis Isolated in Granary Dusts (저곡창고에서 분리된 Bacilus thuringiensis의 특성조사)

  • 김호산;박현우;이대원;유용만;유용만;강석권
    • Korean journal of applied entomology
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    • v.34 no.3
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    • pp.243-248
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    • 1995
  • In order to isolate naturally occurring novel Bacillus thuringiensis strains, we investigated the distribution of B. thuringiensis from granary which exist in Kyong-gi province, Korea. A total of 146 strains of B. thuringiensis producing spore and crystal wre isolated. The toxicity of B. thuringiensis isolates was examined against lepidopteran larvae (Bombyx mori), dipteran larvae (Culex pipiens) and coleopteran larvae (Sitophilus oryzae.), respectively. The results showed that a large number of B. thuringiensis isolates from granary dusts were isolated and most isolates wer toxic to lepidopterous larvae. Also, non-toxic B. thuringiensis isolate was common. In order to isolate many B. thuringiensis, therefore, it suggested that granary is favorable locality.

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Induction of Non-Diapause Eggs and Manifestation of Quantitative Characters by Low Temperature Incubation of Eggs in the Silkworm, Bombyx mori L.

  • Singh, Ravindra;Jayaswal, K.P.;Rao, D.Raghavedra;Kariappa, B.K.;Premalatha, V.
    • International Journal of Industrial Entomology and Biomaterials
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    • v.4 no.2
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    • pp.149-153
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    • 2002
  • Induction of non-diapause eggs and manifestation of quantitative characters were studied in different seasons by low temperature incubation of eggs of a low yielding diapause with coloured cocoons silkworm breed, RDI of Bombyx mori. Hundred percent of nandiapause eggs were induced when the egg incubation was carried out at 15$^{\circ}C$ followed by incubation initially at $25^{\circ}C$ and then at $15^{\circ}C$. The diapause-nondia-pause relation was found to be reversible. Analysis of variance study demonstrated significant variation in all the quantitative characters whereas significant variation due to different incubation treatments was observed for larval span, fifth instar larval span, cocoon yield/10,000 larvae by weight, cocoon weight, cocoon shell weight, and cocoon shell ratio. study on hybrid vigour skewed significant heterosis over mid parent value for four economic characters viz., cocoon weight, cocoon shell weight, cocoon shell ratio and filament length in a Fl hybrid between RD1 with diapause eggs and Japanese type bivoltine $NB_{4} D_{2}$.