• Title/Summary/Keyword: Regeneration and fusion

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Intraspecific Protoplast Fusion of Brevibacterium and Intergeneric Protoplast Fusion between Brevibacterium flavum and Corynebacterium glutamicum and the Metabolic Control of L-Lysine Biosynthesis in Improved Bacterial Strains (Brevibacterium flavum의 동종간 및 Corynebacterium glutamicum과의 이속간 원형질체 융합 및 개량균주의 L-Lysine 생합성의 대사제어)

  • Park, Chung;Im, Beon-Sam;Jeon, Moon-Jin
    • Microbiology and Biotechnology Letters
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    • v.15 no.2
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    • pp.104-111
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    • 1987
  • As a trial method of breeding L-lysine producing strains, the intraspecific protoplast fusion bet-ween Brevibacterium flavum ATCC 21528R and Brevibacterium flavum ATCC 21529S and the intergeneric protoplast fusion between Brevibacterium flavum ATCC 21528R and Corynebacterium glutamicum ATCC 13058S were performed. The optimum conditions for protoplast formation of these strains were examined and the effect of plasma expander on regeneration and/or fusion was also observed. Both fusants No. CH23 and No. CH4l showed higher productivity of L-lysine than those of parental cells under the optimum cultural conditions at a rate of 21% and 8.9%, respectively. And, activity of several enzymes in L-lysine biosynthetic pathway including aspartokinase, a rate-limiting enzyme, was determined. Besides, metabolic control mechanism of L-lysine biosynthesis in fusant No. CH23 and in No. CH41 was investigated to compare with that of parental strains.

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Advances in in vitro culture of the Brassicaceae crop plants

  • Park, Jong-In;Ahmed, Nasar Uddin;Kim, Hye-Ran;Nou, Ill-Sup
    • Journal of Plant Biotechnology
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    • v.39 no.1
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    • pp.13-22
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    • 2012
  • Plant regeneration has been optimized increasingly by organogenesis and somatic embryogenesis using a range of explants with tissue culture improvements focusing on factors, such as the age of the explant, genotype, media supplements and $Agrobacterium$ co-cultivation. The production of haploids and doubled haploids using microspores has accelerated the production of homozygous lines in Brassicaceae crop plants. Somatic cell fusion has facilitated the development of interspecific and intergeneric hybrids in sexually incompatible species of $Brassica$. Crop improvement using somaclonal variation has also been achieved. Transformation technologies are being exploited routinely to elucidate the gene function and contribute to the development of novel enhanced crops. The $Agrobacterium$-mediated transformation is the most widely used approach for the introduction of transgenes into Brassicaceae, and $in$ $vitro$ regeneration is a key factor in developing an efficient transformation method in plants. Although many other Brassicaceae are used as model species for improving plant regeneration and transformation systems, this paper focuses on the recent technologies used to regenerate the most important Brassicaceae crop plants.

Physiological Characteristics of Fusants by Interspecific Protoplast Fusion of the Genus Cellulomonas (Cellulomonas 속 종간 원형질 융합체의 특성)

  • Bae, Moo;Lim, Jung-Hwa
    • Korean Journal of Microbiology
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    • v.28 no.1
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    • pp.47-54
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    • 1990
  • In order to investigate physiological characteristics of fusants by interspecific protoplast fusion of the genus Cellulomonas, protoplasts of Cellulomonas flavigena NCIB 12901 and Cellulomonas bibula NCIB 8142 were fused and cell wall regenerated. To give gene maker, C. bibula was treated with 500 ug/ml NTG for 1 hr and arginine requiring auxotrophic mutants were isolated. Protoplasts of the genus Cellulomonas were obtained by treatment with $600{\mu}{\textrm{g}}$/ml lysozyme, and 0.5M sorbitol was optimal for osmotic stabilizer on protoplast fromation. Protoplast fusion was enhanced by 40% PEG)M.W.6,000) containing 25 mM $CaCl_{2}$ at $30^{\circ}C$ for 30 min and fusion frequency between C. bibula and C. flavigena was $5\times 10^{-4}$. Processes of protoplast formation, cell wall regeneration and protoplast fusion were obsdrved by scanning electron microscope. By comparing enzyme activities of cellulase, exocellobiohydrolase, .betha.-glucosidase of the parent strains of Cellulomonas with those of thier mutants and fusants, fusants with increased enzyme activity were obtained. By the studies on nutritional requirement, antibiotic resistance, cellulolytic enzyme activities, type of peptidoglycan and motility of two mutants and fusants, fusants were proved to be recombinant of both mutant strains.

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Effect of water extract of Danshen on bone regeneration of rat calvarial defect model (랫드 두개골 결손부에서 단삼 수용성 추출물의 골형성 효과)

  • Shim, Kyung Mi;Kim, Se Eun;Kang, Seong Soo
    • Korean Journal of Veterinary Research
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    • v.50 no.3
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    • pp.171-177
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    • 2010
  • The purpose of this study was to evaluate the osteogenic capacity of water extract of danshen (Salvia miltiorrhiza Bunge). We have established in rat critical-sized calvarial defect model using the combination with collagen scaffold and danshen hydrophilic extract. All rats were extinguished at 8 weeks after bone graft surgery, and the bone regeneration ability of bone grafting sides was evaluated by plain radiography and micro-CT. These results revealed water extract of danshen had the potential to promote osteogenesis especially continuous oral administration with local treatment compared to one-shot local treatment. This compound may provide a new alternative agent for growth factors to promote bone healing and bone regeneration. In conclusion, these results suggest that danshen hydrophilic extract have the potential to promote osteogenesis in bone defects. Further studies about fusion technology with salvianolic acid B, peptides, growth factors, and scaffolds using of the combination of tissue engineering, cell engineering and mechanical engineering are needed.

High-frequency Plant Regeneration from Cultured Flower Bud Receptacles of Allium hookeri L.

  • Koo, Ja Choon
    • Horticultural Science & Technology
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    • v.32 no.5
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    • pp.694-701
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    • 2014
  • Allium hookeri L. (Alliaceae family) is an important ethnomedicinal plant native to the Himalayan region of Asia. The aim of this research was to establish a high-frequency plant regeneration system for in vitro propagation of A. hookeri. Among the tissue types examined, receptacle explants derived from immature flower buds showed the highest regeneration rate of shoots ($93.33{\pm}4.63%$), roots ($76.67{\pm}7.85%$), and calli ($80.00{\pm}7.43%$) when cultured on Gamborg B5 (B5) medium containing $10{\mu}M$ 6-benzylaminopurine (BA) + $1{\mu}M$ naphthalene acetic acid (NAA), $0.5{\mu}M$ BA + $5{\mu}M$ NAA, and $1{\mu}M$ BA + $10{\mu}M$ NAA, respectively. Shoot multiplication was superior when cultured in liquid rather than on solid medium and relatively high concentrations of BA, ranging from 5 to $10{\mu}M$. Efficient bulblet formation following root induction from shoot clumps was achieved with culture in liquid B5 medium containing 7% (w/v) sucrose. Regenerated bulblets were successfully acclimatized to ex vitro conditions with a greater than 95% survival rate. By this method, a maximum of 62 plantlets per receptacle could be propagated within 9 weeks of initial culture. The in vitro propagation system established in this study will promote A. hookeri biotechnology, including large-scale production of healthy and aseptic clones, preserving parental genotypes with desirable traits, and genetic manipulation to enhance medicinal value.

Protoplast fusion of Aspergillus oryzae (Aspergilluis oryzae의 원형질체 융합)

  • 이수연;이주실;이영록
    • Korean Journal of Microbiology
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    • v.27 no.3
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    • pp.216-220
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    • 1989
  • As the bsic study about protoplast fusion of amylolytic fungus Aspergillus oryze and nonamyloytic sugar fermenter, Saccaromyces cerevsisae, the intraspecific protoplast fusion of A. oryzae was carried out and the properties of the obtained fusants were investigated. For protoplast fomation from mycellia of auxotrophs, Novozyme 234 as lytic enzyme was the most effective and optimal pH was determined to be pH 5.5-6.0. When the two types of protoplasts were treated with a fusogen including 30% PEG4000, they fused effectively and most of fusants were heterokaryons. Protoplasts aggregated with 30% PEG4000 after fusion treatment were observed by the microscope. Protoplast regeneration frequency was 1.46 to 13.8% and complementation frequency of fusion was 0.12 to 0.16. Fusant strains had a 1.5-fold DNA content compared to that of parent strain. And amylase activity was intermediate between those of parent strains.

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Effects of Demineralized Bone Particle Loaded Poly(lactic-co-glycolic acid) Scaffolds on the Attachment and Proliferation of Costal Cartilage Cells (탈미네랄화된 골분/PLGA 지지체에서 늑연골 세포의 부착과 성장에 미치는 영향)

  • Cho, Sun Ah;Song, Jeong Eun;Kim, Kyoung Hee;Ko, Hyun Ah;Lee, Dongwon;Kwon, Soon Yong;Chung, Jin Wha;Khang, Gilson
    • Polymer(Korea)
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    • v.37 no.5
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    • pp.632-637
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    • 2013
  • It has been widely accepted that costal cartilage cells (CCs) have more excellent initial proliferation capacity than articular cartilage cells as well as the easiness for isolation and collection. This study demonstrated that CCs might be one of the substitutes for articular cartilage cells by tissue engineered cartilage. Poly(lactic-co-glycolic acid) (PLGA) has been extensively tested and used as scaffold material but it was limited by the low attachment of cells and the induction of inflammatory cells. Base on previous our studies, we confirmed demineralized bone particle (DBP) had the power of the reduction of inflammatory reaction and the stimulation proliferation of cells. We fabricated PLGA scaffold loaded with 10, 20, 40 and 80 wt% DBP and then tested the possibility of the regeneration of cartilage using CCs. Assays of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and scanning electron microscope (SEM) carried out to evaluate the attachment and proliferation of CCs in DBP/PLGA scaffolds. Glycosaminoglycan (sGAG) and collagen contents assay were conducted to confirm the effects of DBP on formation of extracellular matrix. This study demonstrated that DBP/PLGA scaffolds showed significant positive effects on cell growth and proliferation due to the vitality of DBP as well as the possibility of the application of CCs for tissue engineered cartilage.

A Study on the Development of Integrated Utilization Considering Multi Functions of Urban Estuarine Area (대도심 하구역 개발과 보전의 융합이용모형 개발 연구 -낙동강 하구역을 중심으로-)

  • Im, Jung-Hyeun;Choi, Jin-Hyu;Kim, Jun-Ho;Yoon, Han-Sam;Ryu, Cheong-Ro
    • Journal of Fisheries and Marine Sciences Education
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    • v.22 no.4
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    • pp.589-603
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    • 2010
  • The purpose of this study is to develop a integrated(fusion) utilization model aimed at maximizing the development/management functions of urban estuarine area and to present an integrated concept by analyzing the existing document and data. The main points of this study are as follows: 1) The integrated utilization model suggested in this paper goes beyond the existing idea of sustainable utilization and conservation, putting an emphasis on rational decision-making in estuary management and peaceful coexistence between human and nature. 2) Policies for utilization/development/conservation/regeneration of urban estuarine area include the establishment of communication system between human and nature and a safety net considering both human and nature, the support for environment-friendly community development and the importance of preserving valuable ecosystem and wetland habitat. 3) Lastly, this study suggested that the major tasks for the fusion utilization model development are the integrated management of estuary areas, the conservation and preservation of wetland ecosystem, proper utilization of estuary's productivity, and the introduction and action plans of the integrated management model including the best way to address contamination and disaster management.

Adhesion and Proliferation Behavior of Retinal Pigment Epithelial Cells on Hesperidin/PLGA Films (헤스페리딘/PLGA 필름에서 망막색소상피세포의 부착과 증식거동)

  • Lee, So Jin;Kang, Su Ji;Kim, Hye Yun;Lee, Jung Hwan;Kim, Eun Young;Kwon, Soon Yong;Chung, Jin Wha;Joo, Choun-Ki;Khang, Gilson
    • Polymer(Korea)
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    • v.38 no.1
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    • pp.24-30
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    • 2014
  • Retinal pigment epithelium (RPE) plays an important role in maintaining the visual function and the degeneration of the RPE causes several retinal degeneration disease. In order to fabricate the suitable carrier for RPE transplantation, the hybrid poly(lactide-co-glycolide) (PLGA) film with hesperidin was prepared. Hesperidin has an anti-inflammatory and antioxidant characteristics. ARPE-19 was seeded on hesperidin/PLGA film and then, cell proliferation was determined by the MTT assay, and cell adhesion and cell morphology were confirmed by SEM. Also, RT-PCR was performed to confirm the expression of the specific genes, and AEC immunohistochemical staining was performed to determine the expression of RPE65. As a result, we confirmed that attachment, proliferation and phenotype maintenance of RPE cells were more excellent on hesperidin/PLGA film than PLGA film, thereby we were able to confirm the potential applications of hesperidin/PLGA film as tissue engineering carrier for regeneration of retina.

Mathematical Modeling of Scheduling Problems for the Fusion Fuel Cycle (핵융합 공정주기에서의 생산 계획 최적화)

  • Lee, Suh-Young;Ha, Jin-Kuk;Lee, In-Beum;Lee, Euy Soo
    • Korean Chemical Engineering Research
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    • v.58 no.4
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    • pp.596-603
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    • 2020
  • In this study, a mathematical model for optimal operation of the fusion fuel cycle is developed based on scheduling approach. The fusion fuel cycle consists of a system for storing and supplying deuterium and tritium, and receiving and separating process after the fusion reaction. Except that tritium is a radioactive material, most of these processes consist of catalytic reactions and separation process. For these reasons, it is possible to apply scheduling approach which is also widely utilized to chemical plants to derive the optimal operating scenarios. The developed model determined the optimal regeneration cycle to minimize the amount of tritium inside the vacuum pumps. Based on the characteristics of various device in the fusion reactor, the optimal tritium plant operation scenario is evaluated. The formulated model was applied to the actual tokamak scenario and utilized to analyze the fuel flow and balance of ITER fuel cycle.