• 제목/요약/키워드: in vitro selection

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p-Fluorophenylalanine Resistant Cell Line Selection and Enzyme Activity from Diploid and Hapliod calli of Nicotiana tabacum cv. BY4 (담배 (Nicotiana tabacum cv. BY4)의 캘러스로부터 p-Fluorophenylalanine 저항성 캘러스 선발 및 효소활성도 측정)

  • 오승철;소웅영;조덕이;오승용;양덕춘
    • Korean Journal of Plant Tissue Culture
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    • v.28 no.2
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    • pp.69-74
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    • 2001
  • Calli were induced on MS medium supplemented with 0.5 mg/L 2,4-D by using the leaf explants of haploid which were derived from the diploid and haploid of Nicotiana tabacum cv BY4. These calli were subcultured on MS medium with the combination of 2.0 mg/L 2,4-D, 1.0 mg/L kinetin and 0.1 mg/L BAP. Cell propagation of diploid plants were good in a combination of 2.0 mg/L 2,4-D, 0.1mg/L BAP in vitro conditions, suspension cultures were conducted in equal condition. Homogenized suspension cultured cells were smeared 2.0 mL each on MS medium with 0~100 $\mu$M PFP, to select the resistant colony to PFP, and were examined after 10d, 20d and 30d. Measurment of fresh weight of cells after 30d of culture shows that with more concentration of PFP in medium the fresh weight of the cells decreased. In case of diploid, selected callus was the highest in vitro treated with 5 $\mu$M PFP. It was higher than control until 100 $\mu$M PFP. The active degree of catalase was the highest in vitro with 5 $\mu$M PFP but the lowest in vitro with 10 $\mu$M PFP on the other hand, in case of haploid plant, the active degree of peroxidase and catalase was the highest in vitro treated with 50 $\mu$M PFP. It's sure that enzyme active degree of between diploid and haploid had big differences.

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Development of In vitro Systems for Heat Tolerance Screening and Tuberization of True Potato Seeds (감자의 耐暑性 器內檢定 및 眞正種子의 器內塊莖形成體系 開發)

  • 김현준;김화영;유승렬;김병현;김정간
    • Korean Journal of Plant Tissue Culture
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    • v.24 no.2
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    • pp.71-75
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    • 1997
  • For the establishment of early selection of heat-tolerant clones through in vitro tuberization of true potato seeds, different temperature treatments for in vitro tuberization were investigated. The ratios of tuberization at 2$0^{\circ}C$ on var. Superior and DTO-33 treated with 5 mg/L of BAP and 500 mg/L of CCC, were 85% and 92%, respectively. At 3$0^{\circ}C$, the ratio of tuberization on DTO-33 was 37%, which revealed strongly heat-tolerant clone. In culture system of in vitro tuberization, the number of tubers per flask at 2$0^{\circ}C$ on non-subculture incubation was more than that on subculture incubation. The condition of non-subculture and short-day treatment for 4 weeks was good for production as 10.6 tubers per flask, which was very similar to that of long-day treatment. On the other hand, tuber diameter on long-day treatment was greater as 11.2 mm than on short-day treatment. Therefore, in vitro tuberization from true potato seeds could be induced under the condition of long-day treatment at darkness.

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Efficient and Reliable in vitro Regeneration System for Rubus Species as the Basis of Genetic Engineering

  • Kalai Katalin;Meszaros Annamaria;Denes Ferenc;Zatyko Jozsef;Balazs Ervin
    • Journal of Plant Biotechnology
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    • v.7 no.4
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    • pp.241-246
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    • 2005
  • Factors affecting regeneration of different Rubus varieties (blackberry, raspberry and their hybrid) were examined and a reliable regeneration system was established. Media for stock plant maintenance were tested; different explants and media were investigated to find the best circumstances for the regeneration. The effect of the commonly used antibiotics was studied to determine the most suitable one for selection of the transformants. We found that both MS and LS media supplemented by $20\;gL^{-1}$ sucrose are suitable for the stock plant maintenance. The optimal hormone content for the stock plants is $0.125\;mgL^{-1}$ 6-benzylaminopurine (BAP) with $0.01\;mgL^{-1}$ indole-3- butyric acid (IBA). The highest regeneration rate was observed on medium containing MS salts with B5 vitamins complemented with glucose, sucrose, maltose, $10\;gL^{-1}$ each, supplemented with benzylaminopurine riboside (BAR) ($2\;mgL^{-1}$) and indole-3-acetic acid (IAA) ($0.1\;mgL^{-1}$). The regenerated shoots appeared directly from the cut edges, without callus phase. Hygromycin and geneticin proved to be good selection agents for the Rubus explants, but due to their severe effect on the tissues we propose to use marker-free constructions for the transformation.

Effect of Sperm Selection by Glass Wool Filtration and Swim-up on the Fertilizing Capacity of Frozen-thawed Boar Sperm (동결융해 돼지정자의 수정능에 대한 Glass Wool여과법과 Swim-up법에 의한 정자 선별의 효과)

  • 박수봉;고대환;정진관
    • Journal of Embryo Transfer
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    • v.7 no.2
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    • pp.133-136
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    • 1992
  • Glass wool filtration and swim-up method resulted in inreasing to 58.3% and 62.7% of the progressive motility in frozen-thawed boar sperm, compared to 34.2% in the untreated sperm. Glass wool filtration tended to be more successful than swim-up method for the survival sfter incubation of 38.5$^{\circ}C$ for 3h. Sperm recovered by both the swim-up method and the glass wool filtration method were tested in an in vitro fertilization to determine which of the two techniques would yield sperm with high fertilizing capacity. The results indicated that there was a significantly(p

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Plant Regeneration from Turnip (Brassica rapa ssp. rapifera) Organs

  • Gendaram Sarantuya;Bae Chang-Hyu
    • Plant Resources
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    • v.8 no.3
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    • pp.286-292
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    • 2005
  • Shoot induction system was developed in the recalcitrant plant species, Brassica rapa ssp. rapifera by using optimum selection of profit organ, phytohormone combination, seedling age and kind of culture container. Out of in vitro cultured leaf segment, petiole, hypocotyl, and cotyledon with petiole, only cotyledon with petiole derived from 4 day-old seedlings induced multiple shoot. The optimum combination of auxin and cytokinin for the multiple shoot induction was MS medium containing 5mg/L BA and 0.5mg/L NAA. The major factors for multiple shoot propagation were part of plant organ, age of seedling, and ratio of auxin and cytokinin. In addition, shoot regeneration was promoted in the 100ml Erlenmeyer flask compared with the $90mm{\times}20mm$ Petri-dish. The induced shoots formed roots easy on MS medium containing 0.1mg/L IBA and the whole plants were successfully cultivated in soil.

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Transdermal Permeation of Riboflavin in Ointment Bases Using Gums & Enhancers (Gum류의 연고제제와 흡수촉진제가 Riboflavin의 경피흡수에 미치는 영향)

  • 오세영;황성규;김판기
    • Journal of Environmental Health Sciences
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    • v.26 no.2
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    • pp.91-96
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    • 2000
  • We investigated characters of transdermal therapeutic system(TTS) and the skin permeability of that with applying drug delivery system(DDS). Natural gums were selected as material of TTS. The permeation of natural gums ointment containing drug in rat skin using diffusion cell model. Permeation properties of materials were investigated for water soluble drug such as riboflavin in vitro. We used glycerin, PEG 600 and oleic acid as enhancers. Since dermis has more hydration than the stratum corneum, skin permeation rate at steady state was highly influenced when glycerin was used in riboflavin. The permeation rate of content enhancer and drug was found to be faster than that of content riboflavin only. These results showed that skin permeation rate of drug across the composite was mainly dependent on the property of ointment base and drug. All the gum ointment tested showed good safety. Proper selection of the materials which resemble and enhance properties of the delivering drug was found to be important in controlling the skin permeation rate.

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Selection and Characterization of Tomato Plants for Osmotic Stress Tolerance Derived from a Gamma Ray Irradiation (감마선 돌연변이원에 의한 Osmotic 스트레스 저항성 토마토 계통 선발 및 특성)

  • Kang, Kwon Kyoo;Jung, Yu Jin
    • Journal of Radiation Industry
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    • v.4 no.3
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    • pp.247-252
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    • 2010
  • The present study has been performed to select the osmotic tolerant lines using polyethylene glycol (PEG 6000) through an in vitro and in vivo mutagensis with a gamma-ray. During the screening, we selected three mutant lines that seemed to confer elevated osmotic tolerance in high concentrations of PEG 6000. Fruits of these mutants (Os-HK101, Os-HK102 and Os-HK103) were increased to sugar concentration, L-glutamine acid, vitamin C content and lycopine content than those of the wild type. Also the chlorophyll contents were few decreased more in the three mutant lines than the WT plants. Our results suggest that the Os-HK101 is characterized as osmotic stress tolerance considering the sugar concentration and lycopine content. It is expected that the result of this study can be used for breeding more competitive species with respect to contents in sugar or functional chemicals from the selected osmotic resistant lines.

Agrobacterium-mediated transformation of Lycopersicon esculentum (cv. MicroTom) with two pathogen-induced hot pepper transcription factors

  • Seong, Eun-Soo;Oh, Sang-Keun;Eunsook Chung;Park, Doil
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.79.1-79
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    • 2003
  • Two pathogen-induced hot pepper transcription factors (CaNACl and CapIfl) were introduced into‘MicroTom’tomato by Agrobacterium tumefaciens-mediated transformation. We used to nptII containing kanamycin resistance gene as a selection marker. Both transformed and non-transformed plants were transferred to pot after rooting test in vitro. To approximate the levels of caNACl transcript in leaves of wild-type and transgenic plants, RNA blots were hybridized with double-stranded full-length CaNACl probe at moderate stringency, Although the relative signal strength for hybridization fluctuated among the samples on different blots, transgenic plant lines N-1, N-2 and N-3 consistently displayed increased levels of CaNACl transcript relative to other transgenic lines and wild-type plants. Of all the transgenic lines examined, line N-7 had the least amount of CaNACl transcript. Role of these transcription factors in pathogen defense will be examined by overexpression in tomato.

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Aptamers (nucleic acid ligands) for trypsin-like serine proteases

  • Gal, Sang-Wan;Jeong, Yong-Kee;Satoshi Nishikawa
    • Journal of Life Science
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    • v.12 no.1
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    • pp.14-18
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    • 2002
  • Subpopulations of nucleotides that bind specifically to a variety of proteins have been isolated from a population of random sequence RNA/DNA molecules. Roughly one in $10^{13}$ random sequence RNA/DNA molecules folds in such a way as to create a specific binding site for small ligands. Since the development of in vitro selection procedure, more than 50 nucleic acid ligands (aptamers) have been isolated. These molecules are very useful for the study of molecular recognition between nucleic acid and protein/organic compound. In addition to these basic studies this method gives us a dream to produce new drugs against several diseases. We focused on several aptamers which specifically binds to trypsin-like serine proteases (thrombin, human neutrophil elastase, activated protein C and NS3 protease of human hepatitis C virus) and want to introduce their structural characteristics and some functions.

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Aptamer-based optical switch for biosensors (압타머 광학 바이오센서)

  • Lee, Joo-Woon;Cho, Jeong Hwan;Cho, Eun Jeong
    • Analytical Science and Technology
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    • v.27 no.3
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    • pp.121-139
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    • 2014
  • In this review, we will discuss aptamer technologies including in vitro selection, signal transduction mechanisms, and designing aptamers and aptazyme for label-free biosensors and catalysts. Dye-displacement, a typical label-less method, is described here which allows avoiding relatively complex labeling steps and extending this application to any aptamers without specific conformational changes, in a more simple, sensitive and cost effective way. We will also describe most recent and advanced technologies of signaling aptamer and aptazyme for the various analytical and clinical applications. Quantum dot biosensor (QDB) is explained in detail covering designing and adaptations for multiplexed protein detection. Application to aptamer array utilizing self-assembled signaling aptamer DNA tile and the novel methods that can directly select smart aptamer or aptazyme experimentally and computationally will also be finally discussed, respectively.