• Title/Summary/Keyword: 누에

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Construction of the cDNA Library from Bombyx mori Larvae and Analysis of the Partial cDNA Sequences (누에 유충의 cDNA 유전자 은행 제작 및 cDNA 클론의 부분염기서울 분석)

  • 김상현;윤은영
    • Journal of Sericultural and Entomological Science
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    • v.38 no.1
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    • pp.13-18
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    • 1996
  • To secure the genetic resources of silkworm, Bomyx mori, the cDNA library was constructed with mRNA isolated from fifth instar larvae. Titer of the cDNA library was about 1.3 X 106 plaques in total. We presumed that the titer covered all transcripts existed in Bombyx mori. Meanwhile, it is knowen that partial cDNA sequences, Expressed Sequence Tags(ESTs), have a good value for the discovery of novel genes and the elucidation of their structures. For this purpose, partial cDNA sequencing was carried out from randomly selected cDNA clones in the library. Partial cDNA sequences of 37 clones were determined and an average of 212 nucleotides of sequence can be read from the clone. The ESTs were searched in GenBAnk database and fifteen ESTs showed significant similarities to enlisted sequences. They included the genes of storage protein, heat shock protein, actin, catalase and so forth. We presumed that the 22 unmatched ESTs were novel genes.

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Isolation and Characterization of Inducible Genes from Bombyx mori Injected with E. coli by Differential Screening (누에에의 차별화 선별을 통한 면역 관련 유도 유전자의 분리와 특성)

  • 김상현;제연호
    • Journal of Sericultural and Entomological Science
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    • v.38 no.1
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    • pp.19-24
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    • 1996
  • To investigate the genes which is related to immune reaction of Bombyx mori, differential screening was carried out using naive and induced B. mori mRNA probe. To begin with, we constructed the cDNA library with mRNA isolated from fifth instar larvae injected with E. coli(4 X 106 cells/larva) using Uni ZAP XR vector kit. Thirty-two inducible cDNAs showing higher intensity on the induced mRNA probing membranes were selected. Partial nucleotide sequences of 29 clones were determined and their expessed sequence tags (ESTs) were produced. Nineteen ESTs in 29 ESTs were matched in GenBank database and the rest of them were found to be unknown. These unmatched ESTs were presumed to be novel genes. The nineteen ESTs contained variable genes related to biological process in Bombyx mori and four classes immune genes. Four clones, BmInc 6, 8, 18 and 27 were similar to two antibacterial peptide genes, hemolin gene and transferrin gene, respectively.

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Morphological and Biochemical Characterization of the Chorion in Interspecific Hybrid Between Bombyx mori and Bombyx mandarina (집누에(Bombyx mori)와 멧누에(Bombyx mandayina)의 종간교잡에 있어서 란각구조 및 Chorion 단백질)

  • 김종길;노시갑
    • Journal of Sericultural and Entomological Science
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    • v.36 no.1
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    • pp.30-36
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    • 1994
  • choChorion(egg-shell) morphology of the F1 hybrid between Bombyx mori and Bombyx mandarina has been observed by scanning electrom microscope and chorion protein was analyzed by electrophoresis. The chorion surface structure of F1 hybrids in the lateral(flat) region was similar to that of maternal line. The F1 hybrids chorion was found to have basically a three layer structure. The middle and inner layer were very much like those of the Bombyx mandarina and Bombyx mori. There were many conic pillar structures in the outer layer of the F1 hybrid, which was similar to Bombyx mandarina. This conic pillar structure had a thin cover layer was more clear in the dorsal and ventral side of the F1 hybrid chorion. The conic pillar structure of Bombyx mandarina was found to be dominant in F1 hybrid chorion irrespective of their maternal line. Major components of chorion protein were analyzed by two-dimensional electrophoresis and found to have isoelectric points in the range of pH 4.0-6.5 and molecular weight 10 to 50 kd. F1 hybrid chorion protein components related directly to those of the maternal line. The conic pillar structure was dominat characteristic and it was present in all F1 hybrid.

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Identification of a Pathogenic Bacterium, Staphylococcus gallinarum, to Bombyx mori (누에(Bombyx mori) 무름병을 야기하는 병원성 세균 Staphylococcus gallinarum의 동정)

  • 김길호;박영진;김용균
    • Korean journal of applied entomology
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    • v.41 no.4
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    • pp.279-284
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    • 2002
  • Flacherie symptom was found in the fifth instar larvae of silkworm, Bombyx mori. The bacterial pathogen was isolated from the hemolymph of the infected silkworm and identified. The isolated bacteria caused a significant flacherie pathogenicity to the fifth instar larvae of B. mori when $5{\times}10^{6}$ cfu (colony-forming unit) of the bacteria was injected into each larva. The infected larvae began to die at 6 days after injection and resulted in complete mortality at 10 days. The bacterium was identified as Staphylococcus gallinarum based on the morphological and physiological characteristics described in Bergey's manual. This identification was further supported by the characters of carbohydrate utility analyzed from a bacterial identification system ($MicroLog^{\circledR}$) and also by the molecular structure of 165-23S rDNA internal transcribed spaces. As an insecticidal action, S. gallinarum caused hemolymph septicemia by its cytotoxic effect on the hemocytes of B. mori.

Study on analysis of components and artificial cultural practice on several culture media of Paecilomyces japonica (눈꽃동충하초 배지별 인공재배법과 성분분석에 관한연구)

  • 이희덕;김용균;김홍규;이가순
    • Korean Journal of Plant Resources
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    • v.12 no.2
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    • pp.102-106
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    • 1999
  • This experiment was carried out to find the method for mass production by artifical cultivition and to analyze the components of Paecilomyces japonica according to several media. Time of inoculation of the Paecilomyces japonica using silkworm was on first day of five molting and infection rate was 72.0%. Optium medium for mass production of the Paecilomyces japonica was known effective for increasing dry weight and fruitbody at brown rice 80g plus pupa powder 20g. Dry weight of Paecilomyces japonica using fungus of silkworm was 1.2g including pupa and length of fruitbody was appeared 3.0cm to 3.5cm. Content of $\beta$ - glucan was very high as 40.5% at inoculation on the first day of the five molting while 16.4% at brown rice, 20.7% at pupa, 23.1% at brown rice plus pupa powder, and 28.7% at pine sawdust plus wheat bran. Mycelium was poor and pinkly conidiospore was formed on media of centipede, maggot and powder of silkworm.

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Expression of Attacin-like Antibacterial Protein NUECIN in Pleurotus ostreatus

  • Kim, Beom-Gi;Yun, Eun-Young;Yoo, Youngbok
    • Journal of Mushroom
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    • v.9 no.3
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    • pp.91-95
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    • 2011
  • Pleurotus ostreatus, the oyster mushroom, is one of the most important edible mushrooms. It is especially susceptible to bacterial blotch disease, which is caused by Pseudomonas tolaasii. In order to develop bacterial blotch disease-resistant transgenic mushroom, NUECIN cDNA, a gene for an antibacterial peptide cloned from Bombyx mori, was overexpressed in Pleurotus ostreatus. NUECIN cDNA was fused to the ${\beta}$-TUBULIN promoter of oyster mushroom and co-transformed with the pTRura3-2 vector into the uracil auxotrophic mutant strain. Twelve transformants containing the NUECIN gene were identified by genomic PCR and Southern blot analysis. NUECIN gene expression was confirmed by Northern blot analysis. Three transformants showed the transcriptional expression of the gene. However, we could not detect expression of the protein in the transformants. This study showed the possibility of transgenic mushroom development for disease resistance.

Varietal Difference in Amylase Activity of Larval Digestive Fluid of the Silkworm, Bombyx mori, reared on Artificial Diet (인공사료로 사육한 누에의 소화액Amylase활성에 있어서 품종간관 차이)

  • 문재유;설광렬
    • Journal of Sericultural and Entomological Science
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    • v.24 no.2
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    • pp.73-80
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    • 1983
  • 1. The varietal difference in amylase activity of the digestive fluid of the 5th instar larvae reared on the artificial diet was investigated, using the parent commercial silkworm varieties of Japanese strain. The amylase activity was large different among silkworm varieties. The activity was strong in Hansaeng-1, Jam 115 and Jam 117, medium in Hansaeng-3, Jam 113, Jam 119, and Jam 201, weak in Jam 107, Jam 121 and Gyeongchu. The amylase of the digestive fluid of ten parent commercial silkworm varieties is possible-(ae) type, compared with +(+$\^$ae/) type of Daizo. 2. To investigate the effect of a-amylase pre-treatment of the artificial diet, larvae were fed with the diet treated by a-amylase during 4th-5th instar periods. The blood sugar content and cocoon qualities were slightly higher in the experimental larvae than those in the control, while showing the slight less body weight, amylase activity and dietary efficient.

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Development of polymorphism genetic marker for identification of the silkworm races (누에 피브로인 유전자 다형성 마커 개발)

  • Choi, Kwang-Ho;Kim, Seong-Ryul;Kang, Seok-Woo;Piao, Yulan;Kim, Sung-Wan;Kim, Kee-Young
    • Journal of Sericultural and Entomological Science
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    • v.53 no.2
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    • pp.124-129
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    • 2015
  • We have previously characterized the complete fibroin light chain gene from one of the silkworm race Baegokjam (Bombyx mori) and found two variable regions (FibL1, intron 2 ~ 3; FibL2, intron 6) with the primer sets designed to cover these variable regions. In this study, we tested the utility of these variable regions as genetic markers for classifying silkworm races. For the purpose, Europian races (Q, PK), Chinese races (C26, C31), Japanese races (N15, N9), and tropical races (SA2, SA5) were used in this experiment. The polymorphism of the FibL1 and FibL2 were divided into two and three types, respectively. The combination of the FibL1 and FibL2 polymorphisms were constant within the same races. The result suggest that the primer sets designed from two variable regions of fibroin light chain gene may be useful as the genetic markers for silkworm races.

Antibacterial effect of natural extracts on oral microorganisms (천연추출물이 구강미생물에 미치는 항균 효과)

  • Hwang, Hye-Jeong;Kim, Min-Young;Kang, Kyung-hee
    • Journal of the Korea Convergence Society
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    • v.13 no.5
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    • pp.97-102
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    • 2022
  • In this study, the antibacterial effect of silkworm extract and Momordica charantia extract on S. mutans and F. nucleatum was investigated. S. mutans or F. nucleatum and 0%, 2%, 4%, 8% concentration of silkworm powder or Momordica charantia powder extract were added to the BHI liquid medium and cultured, and the sample was measured at an optical density of 600nm. As a result, S. mutans, the absorbance value was significantly decreased only in 8% of the silkworm extract, and the absorbance value was significantly decreased in all concentrations in the Momordica charantia extract. The absorbance of F. nucleatum was significantly decreased in a concentration-dependent manner in both the silkworm extract and the Momordica charantia extract. This suggests the possibility that silkworm extract and Momordica charantia extract can be used as materials for the prevention and treatment of oral diseases.

Study on the Inoculation Augmentation of paecizomyces japonicus to the Silkworm, Bombyx mori, Using Dexamethasone (Dexamethasone을 이용한 누에(Bombyx mori)에 대한 동충하초균 (Paecilomyces japonicus)의 접종율 제고에 관한 연구)

  • 김길호;박영진;김용균;이영인
    • Korean journal of applied entomology
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    • v.40 no.1
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    • pp.51-58
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    • 2001
  • Entomopathogenic fungus, Paecilomyces japonicus, has been commercially used as medicinal purpose . The silkworm, Bombyx mori, as an optimal host for the fungi, has been selected and used for the production of the fungal fruit bodies. In current method, newly molted fifth instal larvae should be exposed to the adverse stress environment of high temperature (3$0^{\circ}C$), high relative humidity ( 90%), and starvation for 24h for better fungal inoculation to the host insects. In this study, an alternative method using chemical agent, dexamethasone (DEX: an eicosanoid biosynthesis inhibitor), was tried to get the immunodepressive effect on the larvae to elevate the inoculation rate of the fungi to the silkworm without any harsh rearing environment. DEX (100$\mu\textrm{g}$) showed significantly synergistic effect on the hemocyte lethality of the fungus, and was effective to decrease cellular immune responses measured by the number of hemocyte microaggregation and phenoloxidase activity of the fifth instar larvae in response to the fungal injection. A detergent of 0.05% Triton-X was effective to increase the in- oculation rate of the fungi to the larvae and used in all fungal spraying solutions. Without any environ- mental stress treatment, only DEX (100$\mu\textrm{g}$) injection to the fifth instar larvae followed by the fungal spray was effective to get the inoculation rate equivalent to the current fungal spray method requiring harsh rearing environment. These results suggest that the inoculation of P. japonicus can be elevated by the help of DEX and that the silkworms use eicosanoids to elicit cellular immune response against fungal pathogen.

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