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유전자총 및 Agrobacterium을 이용한 Bentgrass의 형질전환

  • 임용우;김기용;정영수
    • Proceedings of the Korean Society of Grassland Science Conference
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    • 1999.06a
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    • pp.74-74
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    • 1999
  • Bentgrass의 형질전환식물 생산을 위하여 유전자총 (Gene-gun) 및 Agrobacterium기법을 이용하여 형질전환을 시도한 결과를 요약하면 다음과 같다. 1. 캘러스의 유도 및 증식 : Bentgrass (Agrostis palustris)의 종자를 MS-5 배지 (MS 기본배지, 2,4-D 5mg/L, casein hydrolysate 2g 포함)에 치상하여 캘러스를 유도하였다. 유도된 캘러스는 증식을 위하여 MS-3 배지 (MS기본배지, 2,4-D 3mg/L, casein hydrolysate 2g 포함)에서 2주 간격으로 subculture 하였다.(중략)

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Establishment of a Method for the Analysis of Diarrhetic Shellfish Poisoning by Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS를 이용한 설사성패류독소의 분석조건 확립)

  • Lee, Ka-Jeong;Suzuki, Toshiyuki;Kim, Poong-Ho;Oh, Eun-Gyoung;Song, Ki-Cheol;Kim, Ji-Hoe
    • Korean Journal of Food Science and Technology
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    • v.41 no.4
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    • pp.458-463
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    • 2009
  • To establish and validate a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the rapid and accurate quantitation of diarrhetic shellfish poisoning (DSP) toxins, we compared the results from different mobile phases and columns used for their analysis and collision energies for MS/MS experiments. Clear peaks of okadaic acid (OA) and dinophysistoxin-1 (DTX1) were obtained by using a mobile phase comprising aqueous acetonitrile containing 2 mM ammonium formate and 50 mM formic acid. The collision energies were optimized to facilitate the most sensitive detection for each toxin, namely, OA, DTX1, pectenotoxin-2 (PTX2), or yessotoxin (YTX). Further, the maximum ion response was obtained at a collision energy of 48 V for OA and DTX1. We compared the analytical performance of $C_8$ and $C_{18}$ columns. A wide range of toxins namely, OA, DTX1, PTX2, and YTX, except DTX3, were detected by both the columns. Although DTX3 was only detected by the $C_8$ column, we found that the $C_{18}$ column was also suitable for the quantitation of OA and DTX1 the toxins responsible for inducing diarrhea. The limit of detection of OA and DTX1 by the established LC-MS/MS conditions was 1 ng/g, and the limit of quantitation of the toxins under the same conditions was 3 ng/g. The process efficiencies were 91-118% for oysters (Crassostrea gigas) and 96-117% for mussels (Mytilus galloprovincialis) further, we observed no significant effect of matrix during the ionization process in LC-MS/MS. The comparison between mouse bioassay (MBA) and LC-MS/MS yielded varying results because low concentrations of OA and DTX1 were detected by LC-MS/MS in some shellfish samples, which provided positive results on MBA for DSP. The analysis time required by MBA for DSP analysis can be reduced by LC-MS/MS.

Effect of Degradative Products on Dibenzothiophene Desulfurization Activity of Gordona sp. MS6 (Gordona sp. MS6균주의 dibenzothiophene 탈황능에 미치는 분해산물의 영향)

  • 조경숙
    • Korean Journal of Environmental Biology
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    • v.18 no.1
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    • pp.33-39
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    • 2000
  • Gordona sp. MS6, desulfurizing petroleum, can convert dibenzothiophene (DBT) to 2-hydroxybiphenyl (2-HBP) and sulfate. In this study, the effect of DBT degradation product on DBT desulfurization activity was investigated. When $Na_2$SO$_4$ and DBT were simultaneously added in the medium sulfur sources, specific growth rate of strain MS6 was doubled compared to not adding $Na_2$SO$_4$. But, sulfate inhibited DBT desulfurization rate, furthermore, when 0.05 g/L $Na_2$SO$_4$ was supplied DBT desulfurization rate decreased down to 40%. When 2-HBP and its derivative, 2, 2'-dihydroxybiphenyl (DHBP) were not added, DBT desulfurization rate was 5.0 $\mu$mol L$^{-1}$ㆍh$^{-1}$. With the increase of 2-HBP and DHBP concentration, DBT desulfurization rate was decreased. No DBT desulfurization activity of Gordona sp. MS6 was observed when initial concentration was over 0.15 mM 2-HBP and 0.8 mM DHBP [Gordona sp., Desulfurization, Dibenzothiophene, 2-Hydroxybiphenyl, Sulfate].

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31P NMR and ESI-MS Study of Fenitrothion-Copper Ion Complex: Experimental and Theoretical Study

  • Choi, Ho-June;Yang, Ki-Yull;Park, Jong-Keun;Koo, In-Sun
    • Bulletin of the Korean Chemical Society
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    • v.31 no.5
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    • pp.1339-1342
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    • 2010
  • $^{31}P$ NMR and ESI-MS studies of $Cu^{2+}$ binding to Fenitrothion (FN) were performed by experimentally and theoretically. The calculated $^{31}P$ NMR chemical shifts for FN-$Cu^{2+}$ complexes are in good agreement with experimental chemical shifts in order, and the results present an important information for organophosphorus pesticide metal complexes. ESI-MS and low energy CID MS/MS experiments of FN-$Cu^{2+}$ complexes combined with accurate mass measurements give insight into the metal localization and allow unambiguous identification of fragments and hydrolysis products.

Ultra-sensitive Determination of Salinomycin in Serum Using ICP-MS with Nanoparticles

  • Cho, H.K.;Lim, H.B.
    • Bulletin of the Korean Chemical Society
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    • v.35 no.11
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    • pp.3195-3198
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    • 2014
  • An ultra-sensitive detection method for small molecules such as antibiotics was developed using ICP-MS with magnetic and $TiO_2$ nanoparticles. Since most of the antibiotics are too small to employ a sandwich-type extraction through an immunoreaction, a non-specific platform was employed, in which the target was extracted by magnetic separation, followed by tagging with $TiO_2$ nanoparticles of 11.2 nm for ICP-MS measurement. The detection limit for salinomycin obtained from spiked serum samples was $0.4ag\;mL^{-1}$ (${\pm}10.3%$), which was about $1.5{\times}10^6$ times lower than that of LC-MS/MS and about $1.2{\times}10^{11}$ times better than that of ELISA. Such an excellent sensitivity enabled us to study the toxicity of antibiotics exposed to human beings by determining them in serum.

Somatic Embryogenesis and Plant Regeneration in Suspension Cultures of Aralia elata S (두릅캘러스의 현탁배양에서 체세포배발생과 식물체 재분화)

  • 장한호;박철호;이윤수;신영범
    • Korean Journal of Plant Tissue Culture
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    • v.21 no.3
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    • pp.167-171
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    • 1994
  • This study was carried out to investigate the possibility of plant regeneration through somatic embryogenesis in suspension culture of Aralia elata S. Callus was induced from the explants of leaf and petiole cultured in the MS media containing 2,4-D and TDZ. More embryogenic calli were formed from petiole and with combination treatment of 24-D and TDZ. The quarter strength MS medium was effective for increasing number of somatic embryos. Mannitol supplemented to the quarter strength MS medium, reduced somatic embryo formation but inositol increased. Normal plantlets(86%) were regenerated from mature somatic embryos in MS basal medium and 50% of those survied when transplanted to the vermiculite in greenhouse.

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Multi-secondary Somatic Embryogenesis and Plant Regeneration from Hypocotyl Cultures of Alfalfa (Medicago sativa L.) (알팔파의 하배축으로부터 다량의 이차 체세포배 발생과 식물체 재분화)

  • Won, S.H.;Lee, B.H.;Kim, K.Y.;Lee, H.S.;Lee, H.J.;Jo, J.
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.19 no.3
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    • pp.273-280
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    • 1999
  • Hypocotyl explants of Medicago saliva cv. Vernal were cultured on Murashige and Skoog (MS) medium supplemented with various combinations of growth regulators. After six weeks of culture, somatic embryos were formed from calli on MS medium containing $4mg/{\ell}$ 2,4-D and $0.1mg/{\ell}$ kinetin, or $4mg/{\ell}$ 2,4-D and $0.5mg/{\ell}$ kinetin. The mature somatic embryos were developed to plantlets when subcultured on MS basal medium. In order to obtain secondary somatic embryogenic calli, cotyledon of regenerated plantlets were cultured on a callus induction medium. Embryogenic calli were formed on MS medium containing $4mg/{\ell}$ 2,4-D alone. For induction and development of secondary somatic embryogenesis, the embryogenic calli were transferred to MS basal medium containing either 2,4-D or NAA. Multi-secondary somatic embryogenesis was the most effective on MS basal medium with $0.1mg/{\ell}$ 2,4-D. The rate of secondary somatic embryo formation of regenerated plants was 18 times higher than that of seed grown plants. The mature secondary somatic embryo were germinated into plantlets on MS basal medium after six weeks of culture.

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Survey and method validation of simultaneous quantitative analysis of T-2 and HT-2 toxins in cereals (곡류 중 T-2 및 HT-2 독소 동시 정량분석의 유효성 검증 및 실태조사)

  • Paek, Ockjin;Kang, Teabeom
    • Food Science and Preservation
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    • v.22 no.4
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    • pp.559-566
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    • 2015
  • The aim of this study was to develop an analytical method for determination of T-2 toxin and HT-2 toxin level in cereals and to survey their levels using LC-MS/MS. The T-2 and HT-2 toxins were simultaneously analyzed by electrospray ionization with a positive ion mode and multiple reaction monitoring (MRM) after filteration and immuno-affinity column clean-up. A matrix-matched standard calibration used for quantification and recoveries of T-2 and HT-3 toxins were in the range of $100.6{\pm}7.2%$ and $96.8{\pm}9.4%$, respectively. Limits of detection and quantification of T-2 and HT-2 toxins were estimated to be 0.5 and $1.5{\mu}g/kg$, respectively. Each repeatability (RSRr) of T-2 and HT-2 toxins was determined to be 0.9~6.0%, and 4.9~6.1%, respectively. Total 115 samples cereals were collected from 9 types of cereals for analysis. The positive percentages of T-2 and HT-2 toxins obtained from collected samples were found to be 72% and 80%, respectively. The contamination level of T-2 toxin and HT-2 toxin in cereals were $37.1{\mu}g/kg$, and $5.4{\mu}g/kg$, respectively. Therefore, this study suggests that the developed method could be an useful analytical method to determine the T-2 and HT-2 toxin level in cereals and the present data could be used as a reference to estimate the risk assessment.

Determination of Total Aflatoxins in Foods by Parallelism of ELISA and LC/MS/MS (ELISA-LC/MS/MS 병행에 의한 식품 중 aflatoxins 분석)

  • Kim, Kyeong-Yeol;Nam, Min-Ji;Nam, Bo-Ram;Ryu, Hee-Jung;Song, Jeong-Eon;Shim, Won-Bo;Lee, Soo-Hyung;Chung, Duck-Hwa
    • Journal of Environmental Health Sciences
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    • v.36 no.1
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    • pp.52-60
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    • 2010
  • High performance liquid chromatography (HPLC) and liquid chromatography mass spectrometry (LC/MS) have been widely used to quantify aflatoxins in food, but these methods are expensive, time-consuming, unsuitable for analysis of the routine screening of large sample numbers and require derivatization and high level techniques to perform. The objective of this study is to detect aflatoxins in a large number of foods by a high efficient analytical system of combined enzyme linked immunosorbent assay (ELISA) for screening and LC/MS/MS for confirmation. The samples spiked individually with aflatoxin $B_1$ (0.5 and 1.0 ng/g) and total aflatoxins (10 ng/g) were analyzed by ELISA and LC/MS/ MS, and the recoveries for ELISA and LC/MS/MS were 71.8~119.2% and 70.8~135.3%, respectively. A total of 378 samples (grains, nuts, soybean and fermented soybean foods, pepper and fermented pepper foods) were purchased from the six major cities in Korea and analyzed by ELISA-LC/MS/MS system. Twenty two (5.8%; peanut: 11, pistachio: 2, walnut: 6, almond: 1, pepper powder: 1, pepper paste: 1) out of 378 samples were screened as aflatoxin B1 positive by ELISA, but, 4 (1.1%; peanut: 2, pistachio:1, pepper powder: 1) out of the 22 samples screened were confirmed as aflatoxins positive at levels of 1.02~52.79 ng/g by LC/MS/MS. ELISA-LC/MS/MS system provides a more rapid, accurate and cost-effective method for the detection of aflatoxins in large number of samples.

Educational Peptide Mapping of Protein-based Biopharmaceuticals by using LC-MS/MS (LC-MS/MS를 이용한 단백질 의약품 맵핑 교수법)

  • Kim, Junseok
    • Journal of Practical Engineering Education
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    • v.14 no.2
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    • pp.327-332
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    • 2022
  • This experiment presents a precise analysis method using a mass spectrometer in the biopharmaceutical market, where utility is expanding. Among various techniques for analyzing the protein drug, somatotropin, the peptide fragments through biochemical sample preparation was analyzed by LC-MS/MS characterization. The analysis process was performed by separation analysis using nanoUPLC and MS/MS analysis using Orbitrap. In the case of somatotropin with 21 tryptic peptides, 13 of them were consistent with theoretical predictions within an average of 1 ppm error.