• Title/Summary/Keyword: Cell isolation

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Isolation of protoplast from conidiospore of Trichoderma koningii (Trichoderma koningii의 conidiospore로부터의 원형질체 분리에 관하여)

  • 박희문;홍순우;하영칠
    • Korean Journal of Microbiology
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    • v.21 no.4
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    • pp.213-220
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    • 1983
  • Conditions for isolation of protoplasts from conidiospores of Trichoderma koningii ATCC 26113 were tested. Maximum production of conidial protoplasts was obtained by preincubation of conidiospores on liquid minimal medium for 8 1/2 hrs. and by reaction with cell wall lytic enzyme for 3 hrs. Among effective cell wall lytic enzymes (Driselase, p-Glucuronidase, Novozyme and Zymolyase), Driselase was the most effective one on the production of conidial protoplasts. The production of conidial protoplasts was also enhanced by addition of 2-Deoxy-D-Glucose $(25{\mu}g/ml)$ into liquid minimal medium. Over 70% of the initial swollen conidia, preincubated in liquid minimal medium supplemented with 2-Deoxy-D-Glucose $(25{\mu}g/ml)$, were converted to protoplasts by incubation with 2% (w/v) commercial lytic enzyme Driselase at $28^{\circ}C$ for 3 hrs. The reversion frequency of the conidial protoplasts was about 30 times (25-50%) higher than that of mycelial protoplasts (0.6-1.3%).

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Cytotoxic and antimicrobial deterpene from Anisotome Iyallii

  • Choi, Eun-Young;Yang, Hyun-Ok;Choi, Won-Hyung;Lee, Jeong-Ho;Perry-NigelB;Baek, Seung-Hwa
    • Proceedings of the PSK Conference
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    • 2002.10a
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    • pp.392.2-392.2
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    • 2002
  • Cytotoxic activity against the P388 cell line was seen in a crude extract of Anisotome lyallii. A bioactivity guided isolation led to the isolation of a deterpene. which displayed strong Cytotoxic activity against the P388 cell line (IC50 2.3 $\mu$g/ml), as well as antimicrobial activity against Bacillus subtilis. The structure of de terpene 1 was elucidated by spectroscopic methods. (omitted)

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Embryonic Stem Cell and Nuclear Transfer

  • 임정묵
    • Proceedings of the Korean Society of Embryo Transfer Conference
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    • 2002.06a
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    • pp.19-25
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    • 2002
  • Researches on manipulation pluripotent stem cells derived from blastocysts or promordial germ cells (PGCs) have a great advantages for developing innovative technologies in various fields of life science including medicine, pharmaceutics, and biotechnology. Since the first isolation in the mouse embryos, stem cells or stem cell-like colonies have been continuously established in the mouse of different strains, cattle, pig, rabbit, and human. In the animal species, stem cell biology is important for developing transgenic technology including disease model animal and bioreactor production. ES cell can be isolated from the inner cell mass of blastocysts by either mechanical operation or immunosurgery. So, mass production of blastocyst is a prerequisite factor for successful undertaking ES cell manipulation. In the case of animal ES cell research, various protocol of gamete biotechnology can be applied for improving the efficiency of stem cell research. Somatic cell nuclear transfer technique can be applied to researches on animal ES cells, since it is powerful tool for producing clone embryos containing genes of interest. In this presentation, a brief review was made for explaining how somatic cell nuclear transfer technology could contribute to improving stem cell manipulation technology.

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Screening and Isolation of the Antitumor Agents from Medicinal Plants (I) (생약으로부터 항암성분의 검색 및 분리 (I))

  • Park, Shin-Young;Kim, Jin-Woong
    • Korean Journal of Pharmacognosy
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    • v.23 no.4
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    • pp.264-267
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    • 1992
  • The cytotoxic activity of medicinal plants was screened using A549 human lung cancer cell line. Plant materials were extracted with 80% methanol and fractionated to chloroform and water layers. Each methanol, chloroform, and water extract of thirty-two medicinal plants was tested for cytotoxic activity in A549 cell culture system and the cell viability was measured by SRB assay.

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Isolation and identification of porcine reproductive and respiratory syndrome virus from serum samples collected from swine farms (돼지 농장으로부터 수집한 혈청가검물에서 돼지생식기 호흡기증 바이러스의 분리 및 동정)

  • Kim, Hyun-Soo;Kong, Sin-Koog
    • Korean Journal of Veterinary Service
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    • v.22 no.4
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    • pp.363-370
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    • 1999
  • Isolation of PRRSV was attempted from 646 swine sera collected from swine farms. The MARC-145 cell, which is highly permissive to PRRSV, was used for virus isolation, propagation, IFA test, and VN test. Total 36 cytopathic viruses to MARC-145 cells were isolated. The virus isolates were identified as a PRRSV by the IFA test and VN test using the reference sera prepared by experimental infection of reference PRRSV CNV-1 into 30 day-old pig. In addition to serological conformation, ORF5 of genomic RNA of 6 selected cytopathic viruses were amplified by the RT-PCR. The resulting PCR products were examined by electrophoresis in 1.2% agarose gel. An appropriate bands of about 680bp including the flanking sequence of total 80bp were seen on agarose gel.

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Rapid Isolation of Mitochondrial DNA-Depleted Mammalian Cells by Ethidium Bromide and Dideoxycytidine Treatments

  • Yoon, Young Geol;Oh, Yoo Jin;Yoo, Young Hyun
    • Journal of Applied Biological Chemistry
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    • v.57 no.3
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    • pp.259-265
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    • 2014
  • Mitochondrial DNA (mtDNA)-depleted (${\rho}^0$) cells are often used as mtDNA recipients to study the interaction between the nucleus and mitochondria in mammalian cells. Therefore, it is crucial to obtain mtDNA-depleted cells with many different nuclear backgrounds for the study. Here, we demonstrate a rapid and reliable method to isolate mammalian mtDNA-depleted cells involving treatment with the antimitochondrial agents ethidium bromide (EtBr) and 2',3'-dideoxycytidine (ddC). After a short exposure to EtBr or ddC, followed by rapid clonal isolation, we were able to generate viable mtDNA-depleted cells from mouse and human cells and were able to successfully repopulate them with exogenous mitochondria from platelets isolated from mouse and human blood samples. These mtDNA-depleted cells can be used to characterize the nuclear mitochondrial interactions and to study mtDNA-associated defects in mammalian cells. Our method of isolating mtDNA-depleted cells is practical and applicable to a variety of cell types.

2 GHz Down Conversion MMIC Mixer using SiGe HBT Foundry (SiGe HBT 공정을 이용한 2 GHz Down Conversion MMIC Mixer 개발)

  • S.-M. Heo;J.-H. Joo;S.-Y. Ryu;J.-S. Choi;Y.-H. Nho;B.-S. Kim
    • The Journal of Korean Institute of Electromagnetic Engineering and Science
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    • v.13 no.8
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    • pp.764-768
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    • 2002
  • In this paper, a double balanced gilbert cell MMIC mixer was realized in Tachyonics SiGe HBT technology. The fabricated mixer has 17 dB conversion gain, 9.8 dB noise figure, -4.2 dBm output 1 dB compression point, -27 dBc RF to IF isolation, and the good input, output matching characteristics. It draws 10 mA from a 3 V supply. The simulation and the measured results are closer to each other, which confirms accuracy of the model library and reliability of the process.

Isolation of Flavonol Rhamnosides from Loranthus tanakae and Cytotoxic Effect of Them on Human Tumor Cell Lines

  • Kim, Young-Kyoon;Kim, Young-Sup;Choi, Sang-Un;Ryu, Shi-Yong
    • Archives of Pharmacal Research
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    • v.27 no.1
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    • pp.44-47
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    • 2004
  • Loranthus tanakae Fr. et Sav. (Loranthaceae) is a species of mistletoe, a semiparasitic plant growing on the branches of Quercus and Betula species as host trees. In our ongoing search for bioactive compounds from endemic species in Korea, we have investigated to isolate the chemical constituents responsible for the antitumor effect of the MeOH extract of L. tanakae. The ethyl acetate soluble part of the MeOH extract demonstrated a marginal inhibition on the proliferation of the tumor cell lines such as A549 (non small cell lung), SK-OV-3 (ovary), SK-MEL-2 (melanoma), XF498 (central nerve system), and HCT-15 (colon) in vitro. Thus, the activity-guided isolation procedure upon the ethyl acetate soluble part of the extract has been carried out and finally four flavonoid rhamnopyranosides (1-4) were isolated as active principle. The structures of 1-4 were elucidated by the physicochemical and spectral data as rhamnetin 3-O-$\alpha$-L-rhamnoside (1), quercetin 3-O-$\alpha$-L-rhamnoside (2), rhamnocitrin 3-O-$\alpha$rhamnoside (3), and kaempferol 3-O-$\alpha$-L-rhamnoside (4).