• Title/Summary/Keyword: 겔 전기영동

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Studies on the permeability and dissolvability of cathodic electrolyzed water (음극전리수의 침투력과 용해력 연구)

  • Kim Jum-Ji;Kang Dong-Kyu;Ryoo Kun-Kul;Lee Yoon-Bae;Lee Jong-Kwon;Lee Mi-Young
    • Proceedings of the KAIS Fall Conference
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    • 2005.05a
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    • pp.292-294
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    • 2005
  • 본 연구에서는 전기영동 겔에 대한 음극전리수의 침투력과 녹차성분에 대한 음극전리수의 용해력을 일반 물과 서로 비교하였다. 음극전리수와 증류수로 제조한 CBB-R 염색시약으로 polyacrylamide 겔 상에서 단백질을 다양한 시간 동안 염색한 후 염색강도를 서로 비교하였다. 그 결과 음극전리수로 제조한 CBB-R 염색시약은 증류수로 제조한 CBB-R 염색시약보다 같은 반응 시간 동안에 먼저 단백질을 강하게 염색시켰다. 뿐만 아니라 $25^{\circ}C$에서 음극전리수는 일반 물에 비하여 녹차성분에 대해 극히 탁월한 용해력을 나타내었다.

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Purification and Characterization of a Juvenile Hormong Binding Protein from Whole Body Homogenates of the Wax Moth, Galleris mellonella Final Instar Larvae (꿀벌부채명나방 종령유충에서 유약호르몬 결합단백질의 정제와 특성)

  • 안기흥;전상학;이경로
    • Korean journal of applied entomology
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    • v.37 no.1
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    • pp.59-64
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    • 1998
  • A juvenile hormone binding protein (JHBP) has been isolated from the whole body homogenate of Galleria rnellonella final instar larvae by gel filtration. The isolated protein is homogenous as judged by column chromatography and gel electrophoresis in the presence and absence of denaturing agent. The JHBP has a relative molecular weight of 32 k by denaturing gel electrophresis and 28 k by gel filtration. The protein exhibits a dissociation constant of 3.9 x M for JH 111.

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Identification of Loblolly Pine (Pinus taeda L.) Clones through Isozyme Analysis (동위효소분석(同位酵素分析)에 의(依)한 테다소나무(Pinus taeda L.) 클론의 식별(識別))

  • Ryu, Jang Bal;Na, Chun Soo
    • Journal of Korean Society of Forest Science
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    • v.76 no.4
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    • pp.330-337
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    • 1987
  • Megagametophyte tissues from seeds of 45 loblolly pine clones were subjected to horizontal starch-gel electrophoresis. The resulting gels were tested for activity of five enzyme systems (GOT, SDH, PGM, MDH, and AP). Isozymes observed were under control of 11 loci. All 45 clones could be identified with unique genotypes at above 10 loci.

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Comparative Studies on the Enzymatic Properties of Trypsins from Cat-shark and Mackerel -1. Purifications and Reaction Conditions of the Trypsins- (복상어와 고등어의 Trypsin에 관한 비교 효소학적 연구 -1. Trypsin의 정제와 반응조건-)

  • PYEUN Jae-Hyeung;CHO Deuk-Moon;HEU Min-Soo
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.24 no.5
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    • pp.273-288
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    • 1991
  • To elucidate the physiological and biochemical differences between chondrichthyes and osteichthyes, the properties of the specific digestive enzymes in cat-shark, Cephaloscyllium umbratile, and mackerel, Scomber japonicus, were studied. Homogenous trypsin proved through the disc-electrophoresis, SDS-PAG electrophoresis and gel filtration was obtained from the pancreas of cat-shark by $50-70\%$ saturated ammonium sulphate fractionation, DEAE-Sephadex A-50 column chromatography, benzamidine-Sepharose 6B affinity chromatography and Sephadex G-75-120 gel filtration. Two types of trypsins were also obtained from the pyloric caeca of mackerel by $30-70\%$ saturated ammonium sulphate fractionation and the slightly modified procedure from the method adopted in the purification of cat-shark trypsin. The two trypsins, designated trypsin A and B, were proved their homogeneity by disc- and SDS-PAG electrophoresis and gel filtration. The molecular weights of the trypsins were estimated to be 31,700 for cat-shark trypsin, 30,000 for mackerel trypsin A and 29,000 for mackerel trypsin B by SDS-PAG electrophoresis, but those were estimated to be 21,500 for cat-shark trypsin, 23,700 for mackerel trypsin A and 21,500 for mackerel trypsin B by gel filtration. The trypsins exhibited their optimum conditions at pH 9.0 and on temperature ranged from $45^{\circ}C\;to\;50^{\circ}C$ for cat-shark, and at pH 8.0 and a temperature of $50^{\circ}C$ for mackerel trypsin A and B, respectively. The cat-shark trypsin was stable at pH 10.0 and the temperature below $10^{\circ}C$, whereas the mackerel trypsin A and B, were stable in the range over pH 7.0 to pH 9.0 below $10^{\circ}C$ and at pH 8.0 below $35^{\circ}C$, respectively. The mackerel trypsins were severely inhibited by some heavy metal ions such as $Ag^{2+},\;Cu^{2+}\;and\;Hg^{2+}$ compared to cat-shark trypsin. All of the enzymes were also inhibited by antipain, leupeptin, TLCK(tosyllysine chloromethyl ketone) and SBTI(soybean trypsin inhibitor) remarkably. The inhibitory effects of PMSF(phenylmethane sulphonylfluoride), DFP(diisopropyl fluorophosphate) and benzamidine were indicated that these enzymes belong to serine-proteases.

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Convenient Methods for the Extraction and Discrimination of Water-Soluble Plant Pigments (수용성 식물 색소 추출 및 판별에 관한 간편한 방법)

  • Chung, Sang-Ho;Byun, Young-Ho
    • The Journal of the Korea Contents Association
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    • v.9 no.3
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    • pp.353-360
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    • 2009
  • The use of colorants as additives for foods and drinks is a significant factor to food consumers in determining the acceptability of processed foods. In recent years, the number of previously used artificial colorants/dyes suitable for food use has been drastically reduced as a result of toxicological studies. Therefore, the use of natural pigments such as anthocyanins and betacyanins that were known to have anticancer and antioxidant activities is increasingly required. In this study, the water-soluble plant pigments, anthocyanins and betacyanins, were easily extracted with a very simple method using a few organic solvents such as ethyl acetate, ethyl ether, and chloroform. After the extraction of them, these two major plant pigments could be also simply and rapidly separated and discriminated by a solely one-stepped agarose gel electrophoresis in a citrate buffer (pH 3.0).

Asymmetric Diffusion Model for Protein Spot Matching in 2-DE Image (2차원 전기영동 영상의 단백질 반점 정합을 위한 비대칭 확산 모형)

  • Choi, Kwan-Deok;Yoon, Young-Woo
    • The KIPS Transactions:PartB
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    • v.15B no.6
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    • pp.561-574
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    • 2008
  • The spot detection phase of the 2-DE image analysis program segments a gel image into spot regions by an image segmentation algorithm and fits the spot regions to a spot shape model and quantifies the spot informations for the next phases. Currently the watershed algorithm is generally used as the segmentation algorithm and there are the Gaussian model and the diffusion model for the shape model. The diffusion model is closer to real spot shapes than the Gaussian model however spots have very various shapes and especially an asymmetric formation in x-coordinate and y-coordinate. The reason for asymmetric formation of spots is known that a protein could not be diffused completely because the 2-DE could not be processed under the ideal environment usually. Accordingly we propose an asymmetric diffusion model in this paper. The asymmetric diffusion model assumes that a protein spot is diffused from a disc at initial time of diffusing process, but is diffused asymmetrically for x-axis and y-axis respectively as time goes on. In experiments we processed spot matching for 19 gel images by using three models respectively and evaluated averages of SNR for comparing three models. As averages of SNR we got 14.22dB for the Gaussian model, 20.72dB for the diffusion model and 22.85dB for the asymmetric diffusion model. By experimental results we could confirm the asymmetric diffusion model is more efficient and more adequate for spot matching than the Gaussian model and the diffusion model.

Synthesis of Muscle Proteins During the Differentiation of Cultured Chicken Pectoralis Muscle Cells (培養 鷄胚 筋細胞의 分化에 따른 筋特異 蛋白質의 合成과 젖산탈수소 효소의 活性에 관하여)

  • Ha, Doo-Bong;Im, Wook-Bin;Yoo, Byoung-Je
    • The Korean Journal of Zoology
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    • v.24 no.4
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    • pp.173-188
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    • 1981
  • 근세포의 분화에 있어서의 근특이 단백질의 합성 순서를 구명하기 위하여 계배 근세포를 2$\\sim$9일간 배양하면서 단백질합성야상을 SDS-polyacrylamide 겔전기 영동법, 등전점초점2차원 전기영동법 및 방사자기법으로 분석하였다. Actin은 분화의 초기부터 활발히 합성되어 그 양이 다량으로 축적되나, myosin은 배양 3일째부터 대량 합성되기 시작하였다. Myosin의 대량합성시기는 배양 근원세포가 융합을 활발히 일으키는 시기와 거의 같았다. Myoglobin은 분화초기부터 서서히 합성축적되기 시작하여 배양 5일에서 최대치에 달하였다. Creatine phosphokinase는 배양 3일만에, 그리고 glyceraldehyde dehydrogenase는 6일만에 전기영동상에 검출되었다. Tropomyosin $\\alpha$와 $\\beta$, 그리고 troponin C는 분화초기부터 비교적 다량 합성되고 있었다. 젖산탈수소효소의 활성은 배양 2$\\sim$5일 사이에서 급격히 증가하고 이후 거의 변화가 없었다. 이 효소의 동위효소 조성은 초기 근원세포에서는 $H_4$와 $H_3M$형이 많으나 분화가 진행됨에 따라 $HM_3 와 M_4$형이 서서히 출현하였다. 그리고 배양 5일만에 5종의 동위효소가 모두 검출되었다.

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Synergistic Interaction of Radiation with Pesticide on DNA Damage in Human Lymphocytes as Biological Information for Prevention of Environmental Disaster (환경재해 방지를 위한 생물정보로서의 사람 림프구 DNA 손상에 대한 방사선과 살충제의 상승작용)

  • 김진규
    • Korean Journal of Environmental Biology
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    • v.19 no.1
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    • pp.19-24
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    • 2001
  • Agricultural pesticides may cause certain biological risks since they are widely used to eradicate pests. Agricultural disasters may arise even from the possibility of their synergistic interaction with other harmful enviromnetal factors. The effect of pesticide on radiation-induced DNA damage in human blood lymphocytes was evaluated by the single cell gel electrophoresis (SCGE) assay. The lymphocytes, with or without pretreatment of the pesticide, were exposed to 0-2.0 Gy of $^60 CO$ gamma ray. Significantly increased tail moment, which was a marker of DNA strand breaks in SCGE assay, showed an excellent dose-response relationship. The present study confirms that the pesticide has the cytotoxic effect on lymphocytes and that it shows the synergistic interaction with radiation on DNA damage as well. The results may have a role of providing biological information necessary for the prevention of environmental disaster.

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Apolipophorin-III uptake by the adult testes in the wax moth, Galleria mellonella (꿀벌부채명나방 성충 정소에 의한 아포리포포린-III의 흡수)

  • Yun, Hwa-Kyung
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.17 no.10
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    • pp.199-203
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    • 2016
  • Apolipophorin-III (apoLp-III) was isolated and purified from the last larval hemolymph of Galleria mellonella by KBr gradient ultracentrifugation and gel chromatography (Sephadex G-100). After KBr gradient ultracentrifugation, the lipophorin-free fractions were used as the samples for gel chromatography. The purity of the finally purified apoLp-III was confirmed by SDS-PAGE after gel chromatography. In this study, we found that apoLp-III is taken up into the adult testes in Galleria mellonella. The testes were dissected from day-1 or -2 adults in cold Ringer's solution and used for tissue culture. The protein moiety of apoLp-III was labeled with FITC dissolved in dimethyl sulfoxide (DMSO) at room temperature under conditions of continuous stirring for 1 h. The FITC-labeled apoLp-III was purified with a Sephadex G-25 PD-10 column. The tissues of the adult testes were incubated at room temperature for 30 min with fluorescein isothiocyanate (FITC)-labeled apoLp-III. Fluorescein microscopy and sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis (PAGE) revealed that the adult testis tissues internalize the FITC-labeled apoLp-III. The results showed that apoLp-III is taken up by the adult testes.

Polyacrylamide Gel Electrophoresis of the Cell Proteins from Differentiating Aspergillus niger (검정곰팡이의 분화(分化)에 있어서 폴리아크릴아마이드 겔 전기영동법(電氣泳動法)에 의한 단백질 패턴 연구(硏究))

  • Park, Yong-Ja;Kim, Jong-Hyup
    • The Korean Journal of Mycology
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    • v.8 no.3
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    • pp.159-166
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    • 1980
  • Aspergillus niger van Tieghem was cultured by the method of submerged and synchronized culture for the study of differentiation. Acid-phenol soluble cell proteins of the fungus were extracted from four stages during development. Those acid-phenol soluble proteins were separated by polyacrylamide gel electrophoresis to determine the protein patterns. A new protein band was observed from the pre-sporulation body, color density of the stained protein bands in four tubes differed according to the differentiation stages. The number of protein bands in 10% gels varied from 18 to 16, 17, and 19 according to the course of spore germination stage, conidiophore stage, phialide maturation stage and sporulation stage.

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