• Title/Summary/Keyword: Culture Fusion

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Studies on Development of Antagonistic Microorganism by Cell Fusion - Biological control of disease - ) (세포융합에 의한 신 길항미생물 육종에 관한 연구 - 목초 병해의 생물학적 방제 -)

  • 최기춘;이영환;전우복
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.15 no.1
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    • pp.1-12
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    • 1995
  • This study was to investigate an effective biological control of forage diseases and provide a basic data and a model in improving variety of antagonistic bacteria, with growth promoting effect on forage, through cell fusion. The results obtained were summarized as follows; 1. The antagonistic himbacterium against soil-borne phathogenic fungi Fusarium oxysporum and Rhizoctonia solani was isolated from continuous cropping himsphere soil of forage, and its biological and physiological characteristics were investigated. This bacterium was identified as Bacillus subrilis and named BS 101. Another strain for cell fusion was Bacillus thur ingiensis ssp. kurstaki HD-I(BT 37669) with insecticidal crystal. 2. The auxotropic mutants of BS 101 and BT 37669 were derived after mutagenesis using N-methyl-N'nitro- Nitrosoguanidine(NTG) to give amino acid requirement marker. n e s e auxotropic mutants of BS 101 and BT 37669 were named BS 1013(his-) and BT 69(asp-), respectively. 3. The best protoplast requirement was obtained using DM 3 medium, containing 5% casamino acid, 1 M $MgCI_2$ and 2% bovine semm albumin, to give Fusant 3, 7 and 8. BT toxin gene was not identified with fusants by Southern blotting. However, SDS-PAGE analysis of strains showed various protein patterns among fusants. 4. From the dark culture experiment, growth of forage in inoculated soil with antagonistic bacteria was delayed than that of non-inoculated soil with antagonistic bacteria in each continuous cropping soil and in each sterilized soil. On the other hand, growth duration of forage was different between continuous cropping soil and sterilized soil. 5. Seed germination of Alfalfa, Italian ryegrass and Orchardgrass were significantly improved by inoculation of antagonistic bacteria(p< 0.05).

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Protoplast Formation and Fusion between Anastomosis Groups of Rhizoctonia solani (Rhizoctonia solani 융합균(融合菌) 간(間)의 원형질체형성(原形質體形成) 및 융합(融合))

  • Chung, Hoo-Sup;Kim, Dal-Soo;Ahn, Hee-Suk
    • The Korean Journal of Mycology
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    • v.20 no.1
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    • pp.44-50
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    • 1992
  • The protoplast formation of Rhizoctonia solani in the fast growing anastomosis groups (AGs) 1 and 4, the intermediate AG-2 and AG-5, and the slow AG-3 yielded the most, moderate and the least in that order, respectively. Sclerotia formation varied with AGs. A high yield of protoplasts from AGs was obtained with a combined lytic enzyme system containing cellulase 'Onozuka' R-10, macerozyme R-10 and ${\beta}-glucuronidase$. When 3g (fresh weight) of 30 hr old mycelia was incubated for 3 hr at $32^{\circ}C$ with the enzyme mixture in 0.6 M mannitol, maximum protoplasts were obtained in the five AGs. A protoplast fusion between sclerotia forming AG-1 inactivated with heat and non-forming AG-5 was induced by polyethylene glycol and ${Ca}^{2+}$. Seven fusants obtained were based on characteristics of colony and sclerotium formation on culture plates. The fusants were confirmed by isozyme patterns of esterase and killing reaction between AG-1 and a fusant F1501.

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The Research Methodology of Korean Commercial Arbitration in the Discipline-Fusion Perspective (한국 상사중재의 연구방법론 : 학문융합적 관점을 중심으로)

  • Chung, Yong-Kyun
    • International Commerce and Information Review
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    • v.13 no.2
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    • pp.151-176
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    • 2011
  • The main objective of this study is twofold. The first is to investigate the main currents of Korean commercial arbitration. The second is to investigate the research methodology of Korean commercial arbitration in the perspective of disciplinary fusion perspective. The main findings of Korean commercial arbitration are as follows: first, the incidence of commercial arbitration increased in the Korean Commercial Arbitration Board during the period of 2006-2010, second, the primary causes of trade claims are disputes related with payment and those related with sales contracts, third and finally, the number of countries seeking to resolve disputes through arbitration has increased, including European countries. In this setting, it is necessary to investigate commercial arbitration through the inter-disciplinary perspectives. This study suggests that there are five research methodologies of commercial arbitration. They are legal theory related methodology, institutional methodology, cultural methodology, causal methodology and empirical methodology. Traditionally, the legal profession leads the field of commercial arbitration. However, it is necessary to introduce other elements to the field of commercial arbitration. This study highlights the disciplinary fusion approach among five research methodologies of commercial arbitration.

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Production of Putative Somatic Hybrid of Petunia hybrida and. Nicotiana sanderae by Protoplast Fusion (Petunia hybrida와 Nicotiana sanderae의 원형질체융합에 의한 잠정적 체세포잡종 식물체 생산)

  • 정재동;노영희;최수옥;지선옥
    • Korean Journal of Plant Tissue Culture
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    • v.22 no.2
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    • pp.105-110
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    • 1995
  • The experiment were carried out to obtain a somatic hybrid through protoplast fusion between P.hybrid and N. sanderae. The isoenzyme pattern he chromosome number and the phonotype were observed for genetic study on the regenerants obtained from the fusion product cultures. Putative somatic hybrids possessed all the bands that appeared in both mother plane. A specific band was found on the top of the banding pattern which was assumed to be a marker band of somatic hybrid between two genera. Aspartate amninotransferase isoenzyme bands which were found in both mother plane were also revealed in the putative somatic hybrids or deleted in the upper part of H. sanderae band pattern. The chromosome number of P.hybrida was 2n=14, while N, sanderae was 2n=18,but the number of the putative somatic hybrids ranged from n=32 to 36. The phonotype of putative somatic hybrids was intermediate of the mother plants.

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Fusion Hybrid and Carotenoid Formation from the Yeast, Phaffia rhodozyma (효모 Phaffia rhodozyma의 융합체와 Carotenoid 생성)

  • Chang, Kee-Myung;Kim, Moon-Hee;Song, Myung-Hee;Kim, Sang-Moon;Chun, Soon-Bai
    • The Korean Journal of Mycology
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    • v.21 no.1
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    • pp.9-15
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    • 1993
  • ABSTRACT: The cell fusants were constructed from complementary auxotrophic mutants of Phffia rhodozyma. The nuclear fusion of the fusants was demonstrated by several techniques including comparision of cell volume, estimation of DNA content and nuclear staining. The cell fusants were very stable for succeeding transfer culture on complex medium for more than one year. Malt extract (1%, w/v) and abscisic acid(1 mM) increased the carotenoid formation whereas gibberellic acid(5mM) and riboflavin(0.1 mM) decreased the corresponding content.

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Purification and Characterization of a Subtilisin D5, a Fibrinolytic Enzyme of Bacillus amyloliquefaciens DJ-5 Isolated from Doenjang

  • Choi, Nack-Shick;Chung, Dong-Min;Han, Yun-Jon;Kim, Seung-Ho;Song, Jae-Jun
    • Food Science and Biotechnology
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    • v.18 no.2
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    • pp.500-505
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    • 2009
  • The fibrinolytic enzyme, subtilisin D5, was purified from the culture supernatant of the isolated Bacillus amyloliquefaciens DJ-5. The molecular weight of subtilisin D5 was estimated to be 30 kDa. Subtilisin D5 was optimally active at pH 10.0 and $45^{\circ}C$. Subtilisin D5 had high degrading activity for the A$\alpha$-chain of human fibrinogen and hydrolyzed the $B{\beta}$-chain slowly, but did not affect the $\gamma$-chain, indicating that it is an $\alpha$-fibrinogenase. Subtilisin D5 was completely inhibited by phenylmethylsulfonyl fluoride, indicating that it belongs to the serine protease. The specific activity (F/C, fibrinolytic/caseinolytic activity) of subtilisin D5 was 2.37 and 3.52 times higher than those of subtilisin BPN' and Carlsberg, respectively. Subtilisin D5 exhibited high specificity for Meo-Suc-Arg-Pro-Tyr-pNA (S-2586), a synthetic chromogenic substrate for chymotrypsin. The first 15 amino acid residues of the N-terminal sequence of subtilisin D5 are AQSVPYGISQIKAPA; this sequence is identical to that of subtilisin NAT and subtilisin E.

Production and Purification of Single Chain Human Insulin Precursors with Various Fusion Peptides

  • Cho, Chung-Woo;Park, Sun-Ho;Nam, Doo-Hyun
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.6 no.2
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    • pp.144-149
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    • 2001
  • For the production and purification of a single chain human insulin precursor, four types of fusion peptides $\beta$-galactosidase (LacZ), maltose binding protein (MBP), glutathione-S-transferase (GST), and (His)(sub)6-tagged sequence (HTS) were investigated. Recombinant E. coli harboring hybrid genes was cultivated at 37$\^{C}$ for 1h, and gene induction occurred when 0.2mM of isopropyl-D-thiogalactoside (IPTG) was added to the culture broth, except for E. coli BL21 (DE3) pLysS harboring a pET-BA cultivation with 1.0mM IPTG, followed by a longer than 4h batch fermentation respectively. DEAE-Sphacel and Sephadex G-200 gel filtration chromatography, amylose affinity chromatography, glutathione-sepharose 4B affinity chromatography, and a nickel chelating affinity chromatography system as a kind of immobilized metal ion affinity chromatography (IMAC) were all employed for the purification of a single chain human insulin precursor. The recovery yields of the HTS-fused, GST-fused, MBP-fused, and LacZ-fused single chain human insulin precursors resulted in 47%, 20%, 20%, and 18% as the total protein amounts respectively. These results show that a higher recovery yield of the finally purified recombinant peptides was achieved when affinity column chromatography was employed and when the fused peptide had a smaller molecular weight. In addition the pET expression system gave the highest productivity of a fused insulin precursor due to a two-step regulation of the gene expression, and the HTS-fused system provided the highest recovery of a fused insulin precursor based on a simple and specific separation using the IMAC technique.

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A Study on the Characteristics of Maximalism in Men's wear (남성복에 나타난 맥시멀리즘의 표현 특성 연구)

  • Kim, Ji Hye;Yoo, Youngsun
    • Journal of the Korean Society of Costume
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    • v.65 no.5
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    • pp.1-13
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    • 2015
  • The purpose of this study is to make a contribution to men's fashion by analyzing changes related to excessive and decorative expressions of latest men's wear design in the viewpoint of Maximalism, and using the results to develop a new fashion style. Results of the study are as follows: The characteristics of Maximalism in men's wear collection in the last 5 years can be classified as 'Fusion Maximalism, 'Decorative Maximalism', 'Exaggerative Maximalism', 'Humorous Maximalism', and 'Duplicative Maximalism'. The characteristics of each type of Maximalism can be summarized as follows: First, Fusion Maximalism creates new men's fashion style by fusing designs from different cultures in clothing items as well as dismantling gender boundaries in clothing design, and by having designs which transcends time and space. Second, Decorative Maximalism expresses ambiguous decadence by implementing feminine decoration that applies handicraft technique, as well as exaggeratedly using details and trimmings of contrasting images, and applying women's clothing items. Third, Exaggerative Maximalism represents extreme avant-garde that changes the idea of fashion design by changing the styling or the exaggerated silhouette. Fourth, Humorous Maximalism expresses not only the creation of new men's fashion look, but also expresses humanism with exaggerative enjoyment and pleasant ideas. Fifth, Duplicative Maximalism expresses creative design by implementing vivid images of different period and culture, and reproducing successful images of the past into modern fashion.

Studies on the Isolation, Culture and Fusion of Protoplasts from Plant Mesophyll and Cells Cultured in vitro (식물원형질체의 분리, 배양 및 융합에 관한 연구)

  • Choi, S.J.;Son, S.H.;Chang, W.C.
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.27 no.2
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    • pp.147-154
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    • 1982
  • This study was conducted to investigate an effective method of protoplast isolation, the plating efficiency for cell division, and fusion of plant protoplasts by polyethylene glycol for somatic hybridzation. The effectiveness of protoplast isolation was different with the various enzyme concentrations, but, in the protoplast isolation from tobacco mesophyll, the enzyme solution with 0.5% macerozyme and 2.0% cellulase was very effective. The protoplast isolation from callus cultured in vitro for a long period was not obtained in any of the enzyme solution used. Protoplasts divided actively at cell densities above $10^44/ml and at $25^{\circ}C$ under 12hr illumination by inflorecient light (l50 Lux), regardless of presence of agar. The highest frequency of protoplast fusion was obtained after treatment with a solution of 0.33 M polyethylene glycol 1500.

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Transduced PEP-1-Grb7 Fusion Protein Suppressed LPS-induced COX-2 Expression

  • An, Jae-Jin;Kim, So-Young;Lee, Sun-Hwa;Kim, Dae-Won;Ryu, Hea-Jin;Yeo, Seung-Il;Jang, Sang-Ho;Kwon, Hyung-Joo;Kim, Tae-Yoon;Lee, Sang-Chul;Poo, Ha-Ryoung;Cho, Sung-Woo;Lee, Kil-Soo;Park, Jin-Seu;Eum, Won-Sik;Choi, Soo-Young
    • BMB Reports
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    • v.40 no.2
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    • pp.189-195
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    • 2007
  • Although the incidence and severity of atopic dermatitis (AD) is steadily increasing at an alarming rate, its pathogenic mechanisms remain poorly understood yet. Recently, we found that the expression of Grb7 protein was markedly decreased in AD patients using proteomic analysis. In the present study, human Grb7 gene was fused with PEP-1 peptide in a bacterial expression vector to produce a genetic in-frame PEP-1-Grb7 fusion protein. The expressed and purified PEP-1-Grb7 fusion proteins transduced efficiently into skin cells in a time- and dose-dependent manner when added exogenously in culture media. Once inside the cells, the transduced PEP-1-Grb7 protein was stable for 48 h. In addition, transduced PEP-1-Grb7 fusion protein markedly increased cell viability in macrophage RAW 264.7 cells treated with LPS by inhibition of the COX-2 expression level. These results suggest that the PEP-1-Grb7 fusion protein can be used in protein therapy for inflammatory skin disorders, including AD.