• Title/Summary/Keyword: 누에

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Germ Line Transformation of the Silkworm, Bombyx mori L. with a piggyBac Vector Harboring the Human Lactoferrin Gene (락토페린 유전자도입 piggyBac 벡터에 의한 누에 형질전환)

  • Kim, Yong-Soon;Sohn, Bong-Hee;Kim, Kee-Young;Jung, I-Yeon;Kim, Mi-Ja;Kang, Pil-Don
    • Journal of Sericultural and Entomological Science
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    • v.49 no.2
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    • pp.37-42
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    • 2007
  • Lactoferrin, an ion-binding 80-kDa glycoprotein, has been suggested to have many biologic activities, such as facilitating ion absorption and having antimicrobial and anti-inflammatory effects. Several of these activities are likely to only be facilitated by human lactoferrin because they depend on the binding of human lactoferrin to specific receptor. To produce recombinant human lactoferrin to animal foods using transgenic silkworm, Bombyx mori L, we have cloned and sequenced the cDNA encoding for a human lactoferrin (HLf) from the mRNA in mammary tumor line (GI-101). As a result, the 2.5-kb fragment of HLf gene was cloned with pGEM-T vector and then this fragment was sequenced. In the nucleotide sequence analysis, single open reading frame of the 2,136-bp encoding for a polypeptide of 712 amino acid residues was detected. On the other hand, we constructed a recombinant plasmid(pPT-HLf), containing human lactoferrin gene for germ line transformation of the silkworm using a piggyBac transposon-derived vector. A nonautonomous helper plasmid encodes the piggyBac transposase. Approximately 6.7% of individuals in the G0 silkworms expressed green fluorescent protein (GFP). PCR analyses of GFP-positive silkworms (G0 and G1) revealed that independent insertions occurred frequently. Furthermore, Western blot analysis showed that the recombinant HLf expressed in hemolymph has the same molecular weight (80 kDa) as a native protein. On the basis of these experiments, expression of HLf in next generation of transgenic silkworm is now in process.

Effect of Silkworm Hemolymph Collected Large-scale Bleeding Method in Insect Cell Culture (절피원심법으로 대규모 채취한 누에체액의 곤충세포 증식효과)

  • Choi, Ji-Young;Kim, Jong-Gill;Choi, Young-Cheol;Kim, Sam-Eun;Han, Myung-Sae
    • Journal of Sericultural and Entomological Science
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    • v.49 no.1
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    • pp.28-32
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    • 2007
  • This study was carried out to investigate the utilization of hemolymph of silkworm, Bombyx mori, as a substitute for fetal bovine serum(FBS) in the insect cell culture. Hemolymph is collected on a small scale by clipping the abdominal leg; however, this method is not appropriate for large scale collection. The hemolymph was collected from 5 th instar larva by centrifugation after cutting of the abdominal legs was more appropriate procedure for large scale collection. The cell growth in the medium supplemented with hemolymph(Baekokjam) collected in large scale was almost same as that in the medium hemolymph supplemented with hemolymph collected in small scale. However, the mutant($wE^b$) hemolymph collected in large scale was still less effective in the cell growth, as compared to the Baekokjam hemolymph collected in large scale. The optimum centrifugation condition for large-scale bleeding was 500 rpm and 15 min.

Phylogenetic Relationships and Characterization of Korean Native Silkworm Strains Based on RAPDs and Isozyme Analysis, Bombyx mori (동위효소 및 RAPD분석에 의한 한국재래종 누에계통의 계통학적 특성)

  • 이재만;노시갑
    • Journal of Sericultural and Entomological Science
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    • v.43 no.2
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    • pp.59-66
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    • 2001
  • This study was conducted to elucidate phylogenetic relationships and genetic characterization of silkworms that might be recognized as the Korean native strains. Genetic characterization in isozymes and the proteins of larval hemolymph of 17 silkworms were observed by acrylamide gel eletrophoresis, on 12 genes; Bph, Bes, les, Amy-hc, Ict-A, -B, -D,-E,-H, Pfl, Pst, Lp. Gene frequencies in each locus were compared other geographic strains. Korean native strains were remarkably different from others considered as the genetic characterization of Korean native strains. Phylogenetic relationships in Korean native strains were analysed using RAPD-PCR markers. A total of 40 primers were used and 346 bands of amplified DNA were generated from geographic strains. Genetic similarity based on the RAPD bands was used to construct phylogenetic dendrogram based on analysis of bard sharing data of amplified markers. Genetic similarity ranged from 0.595 to 0.860. In the genetic relationship based on dendrogram, they were classified into Bombyx mori group (including 16 domesticated silkworm strains) and B. mandarina group. The Bombyx mori group was separated into three sub-groups at the genetic similarity of 0.6930, including Korean, Japanese and Chinese groups. According to this result, the Korean native variety can be considered as a clearly different variety from other geographic strains. It may be concluded that the Korean native strains are also one of original geographic variety such as Japanese, Chinese, etc.

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Effect of Diflubenzuron on Silkgland Development of Silkworm, Bombyx mori (Lepidoptera: Bombycidae) (Diflubenzuron이 누에의 실샘 발달에 미치는 영향)

  • 김영섭;손해룡
    • Journal of Sericultural and Entomological Science
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    • v.43 no.2
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    • pp.77-82
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    • 2001
  • This study was investigated to know the effects of diflubenzuron(DFB) on the larval silkgland development of the silk-worm, Bombyx mori (L.). It has been known as a prohibitor of chitin synthesis mostly on the species of the Order Lepidoptera. In this work, the effects of the DFB concentrations (2.5$\times$10$\^$-1/,2.5$\times$10$\^$-2/, and 2.5$\times$10$\^$-3/$\mu\textrm{g}$/$\mu$l on the various larval stages, were investigated in terms on the silkgland. The macro- and microstructure of cell membrane of silkgland, and the differences of silkgland weight and water contents treated by DFB are also surveyed. As the tesults, the silkgland weight depended sensitively on not the DFB concentration but the DFB treatment period. The longer DFB treatment period, the lighter dried silkgland weight and the heavier water content of the silkgland. White opaque(WO) emerged in the middle silkgland of DFB treated larva. From the scanning election microscope observation, the cell membrane of silkgland of DFB treated larva was distinctly more compact compared to that of control. The WO was evidently resulted from the obstacles of normal transformation of silk protein through the cell membrane of middle silkgland.

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Effect of Some Herbal Extracts on Entomopathogenic Nematodes, Silkworm and Ground Beetles (몇 가지 한약재 추출물이 곤충병원성선충과 누에 및 먼지벌레에 미치는 영향)

  • Lee, Dong-Woon;Choi, Hyeon-Cheol;Kim, Tae-Su;Park, Jong-Kyun;Park, Jung-Chan;Yu, Hwang-Bin;Lee, Sang-Myoung;Choo, Ho-Yul
    • Korean journal of applied entomology
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    • v.48 no.3
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    • pp.335-345
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    • 2009
  • Effect of four nematicidal herbal extracts (Daphne genkwa, Eugenia caryophyllata, Quisqualis indica and Zingiber officinale) and 3 acricidal herbal extracts (Pharbitis nil, Xanthium strumarium, and Desmodium caudatum) on entomopathobenic nematodes [Steinernema carpocapsae Pocheon strain (ScP) and Heterorhabditis sp. Gyeongsan strain (HG)], silkworm (Bombyx mori), and ground beetles (Synuchus sp.) were investigated in the laboratory and field. D. genkwa was highly toxic to SCP and HG (100% mortality) at the concentration of 5,000 ppm in X-plate. All the infective juveniles of HG were dead after 3 days by E. caryophyllata and Q. indica. The mortality of ScP and HG was below 10% by D. genkwa, D. caudatum, E. caryophyllata, Q. indica and Z. officinale at the concentration of 1,000 ppm two days after treatment while mortality of HG was 62.8% by D. genkwa at the concentration of 1,000 ppm in X-plate. However, 1,000 ppm had not effect on nematode survival and pathogenicity of ScP in sand column. On the contrary, E. caryophyllata had effect on pathogenicity of HG. Mean number of dead Galleria mellonella larva of HG was 0.5 in E. caryophyllata treatment. Q. indica did not effect silkworm reared on mulberry leaves at the treatment of 1,000 ppm in 10 days after treatment. However, there were 20.0 and 100% mortalities in the treatment of D. genkwa 3 and 10 days after treatment, respectively. The weight of silkworm was low in D. genkwa and did not pupate. The weight of pupa and cocoon were not different in E. caryophyllata, P. nil, Q. indica, X. strumarium and Z. officinale. D. genkwa, E. caryophyllata, P. nil, Q. indica and Z. officinale had no effect on ground beetles, Synuchus sp. in forest soil.

Effect of Neem and Mustard oils on Entomopathogenic Nematodes and Silkworm (Neem과 mustard oil이 곤충병원성 선충과 누에에 미치는 영향)

  • Ha, Pan-Jung;Kim, Tae-Su;Lee, Shin-Hae;Choo, Ho-Yul;Choi, Sung-Hwan;Kim, Young-Sub;Lee, Dong-Woon
    • The Korean Journal of Pesticide Science
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    • v.14 no.1
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    • pp.54-64
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    • 2010
  • Effects of thirteen essential oils (anise oil, clove oil, marigold, mustard oil, neem oil, quassia, quilaja, rosemary oil, rotenone, tea tree extract, thyme oil, wintergreen oil, and yucca) and caffeine on typical industrial insect, silkworm (Bombyx mori) and two entomopathogenic nematodes, Steinernema carpocapsae GSN-1 strain (Sc) and Heterorhabditis sp. Gyeongsan strain (Hg) were investigated in the laboratory. When 1,000 ppm of each essential oils was treated, neem oil showed the highest insecticidal activity against silkworm. Mortality of silkworm fed on neem oil treated mulberry leaf was 55.3 and 100% 5 and 10 days after treatment, respectively. The silkworm fed on neem oil treated mulberry leaf did not make cocoon and pupa. Weight of cocoon and pupa was low in rotenone treatment showing 0.27 g and 1.01 g, respectively. Mustard oil had the highest nematicidal activity against entomopathogenic nematodes. 20 ppm of mustard oil resulted in 69.0% and 100% mortality of Sc and Hg 3 days after treatment, but 4% and 36% at 5 ppm in X-plate, respectively. Mortality of baited Galleria mellonella larva by Sc was not different from control at the concentration of 100 ppm of mustard oil while 30% lower in Hg in sand barrier. Mean numbers of established infective juveniles of Hg in Galleria larva were lower than Sc in sand barrier. Survival rate of Sc was similar to control at the concentration of <200 ppm of mustard oil in sand barrier.

The Effect of Nicotine-Contaminated Mulberry Leaf in the Vicinity of Tabacco Drying Plant on Cocoon Crop (연초건조장 부근의 뽕잎이 잠작에 미치는 영향)

  • 양성열;이상풍;김계명;이상욱
    • Journal of Sericultural and Entomological Science
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    • v.20 no.2
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    • pp.26-31
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    • 1978
  • The objective of the present study was to investigate the effect of nicotine-contaminated mulberry-leaf, which was grown in the vicinity of tobacco drying plant (TDP), on cocoon crop. Mulberry-leaf harvested from the field at Sericultural Experiment Station (SES), Suweon, Korea, was used as control, supposedly nicotine-free leaf. Leaf harvested from the field in the distances of 30-50m, 300-400m and 700-800m from TDP was fed during the whole larval stage of the silkworm at. SES. The effect of leaf in each treatment level on the quantitative characters of the silkworm was summarized as follows; 1. Larval duration from 4th instar on was significantly longer in the TDP-leaf treatments than for the control. 2. Duration of matured silkworm appearance became longer as the distance of the mulberry-field from TDP got shorter, because the larval duration and growth of the silkworm were not uniform in the TDP-leaf treatments. 3. Mortality rates during the late larval, cocoon spinning, and pupal stages were highest for the 30-50m leaf, especially mortality rates during the late larval and pupal stage were serious. 4. Pupation rate was lowest for the 30-50m leaf and those for the 300-400m and the 700-800m leaf were not significantly different from that of the control. 5. Nicotine damage to cocoon weight and cocoon shell weight was significant in each TDP-leaf level. Cocoon shell ratio was reduced at the same extent in each level, compared with the control. 6. The ratio among cocoon-classes was significantly different between treatments, compared with best-cocoon ratio of 87.1% for the control. Cocoons were not uniform for the 30-50m leaf, and those for the 300-400m and the 700-800m leaf were as almost uniform as those for the control. 7. Loss of fresh cocoon yield became greater as the distance of the mulberry-field from the TDP-got shorter. In conclusion, the critical distance of mulberry-field, which influences larval health, cocoon quality and yield, appeared to be 800m from the TDP. Such other factors as wind direction and topographic location may be involved in the critical distance. 8. From the present experiment, we could obtain only the effect of nicotine on the silkworm through digestive system, since the silkworm was raised at SES in Suweon. If the silkworm.. were raised in the vicinity of the TDP, poison effect of nicotine on the silkworm could beo expected through exoskeleton and tracheal system as well as through digestive system.

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