• Title/Summary/Keyword: gel electrophoresis

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Identification and Purification of a Normal Rat Liver Plasma Membrane Surface Protein which Disappears after Chemical Carcinogenesis

  • Kim, Min-Young;Lee, Myung-Kyu;Hahm, Kyung-Soo
    • BMB Reports
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    • v.28 no.6
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    • pp.504-508
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    • 1995
  • The electrophoretic patterns of plasma membrane surface proteins of normal rat liver cells and rat hepatomas were compared in 10% non-denaturing and 7-15% gradient non-denaturing gel. Chemical carcinogens, 2-Me DAB (2-methyl-4-dimethylaminoazobenzene) and DENA (diethylnitrosamine), were used to induce hepatoma in rats. One protein which disappeared in hepatoma was identified in normal rat liver by non-denaturing gel electrophoresis. Rabbit antisera were raised against this specific protein, and the protein was purified by Sephacryl S-200 column and immunoaffinity chromatography using the purified antibody. The purified protein showed two bands of molecular weights approximately 50 $kD_{\alpha}$ and 52 $kD_{\alpha}$ by SDS-polyacrylamide gel electrophoresis, which reacted specifically with the antibody. However only one band was observed in non-denaturing gel and also in isoelectric focusing with a pI value of 6.6. This study showed the existence of an unique protein on the plasma membrane surface of normal rat liver cells which disappeared in rat hepatomas induced by chemical carcinogens.

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Capillary Gel Electrophoretic Analysis of Cattle Breeds Based on Difference of DNA Mobility of Microsatellite Markers

  • Lee, Mi-Ji;Yoon, Du-Hak;Jeon, Jin-Tae;Eo, Seong-Kug;Kang, Seong-Ho
    • Bulletin of the Korean Chemical Society
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    • v.30 no.11
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    • pp.2655-2660
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    • 2009
  • A breed of cattle, i.e., Korean cattle (Hanwoo), was identified based on the DNA mobilities of their microsatellites (MSs) by capillary gel electrophoresis (CGE) with a laser-induced fluorescence (LIF) detector. The MS markers were used for the accurate identification of species-specific genes. The DNA mobilities of the MS markers of Hanwoo and Holstein were measured using a CGE system with a fused-silica capillary (inner diameter of 75 ${\mu}m$, outer diameter of 365 ${\mu}m$, and total length of 50 cm). The capillary was dynamically coated with 1.0% (w/v) polyvinylpyrrolidone ($M_r$ = 1,000,000) and then filled with a mixture of 1.3% (w/v) poly(ethylene oxide) ($M_r$ = 600,000) and 1.9% (w/v) poly(ethylene oxide) (Mr = 8,000,000) as a sieving gel matrix. The species-specific genes of Hanwoo and Holstein were clearly distinguished within 33 min. This CGE assay technique is expected to be a useful analytical method for the fast and accurate identification of breeds of cattle.

Lane Detection and Tracking Algorithm for 3D Fluorescence Image Analysis (3D 형광이미지 분석을 위한 레인 검출 및 추적 알고리즘)

  • Lee, Bok Ju;Moon, Hyuck;Choi, Young Kyu
    • Journal of the Semiconductor & Display Technology
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    • v.15 no.1
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    • pp.27-32
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    • 2016
  • A new lane detection algorithm is proposed for the analysis of DNA fingerprints from a polymerase chain reaction (PCR) gel electrophoresis image. Although several research results have been previously reported, it is still challenging to extract lanes precisely from images having abrupt background brightness difference and bent lanes. We propose an edge based algorithm for calculating the average lane width and lane cycle. Our method adopts sub-pixel algorithm for extracting rising-edges and falling edges precisely and estimates the lane width and cycle by using k-means clustering algorithm. To handle the curved lanes, we partition the gel image into small portions, and track the lane centers in each partitioned image. 32 gel images including 534 lanes are used to evaluate the performance of our method. Experimental results show that our method is robust to images having background difference and bent lanes without any preprocessing.

Polyacrylamide Gel Electrophoresis of Proteins of the Meal-Worm, Ephestia kuhniella Zeller (Polyacrylamide Gel 전기영동법에 의한 밀얼룩명나방의 단백질 분리)

  • Yoo, Chong-Myung;Lee, Kyung-Ro
    • The Korean Journal of Zoology
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    • v.16 no.3
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    • pp.185-191
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    • 1973
  • The soluble whole body proteins of meal-worm Ephestia kuhniella, at various developmental stages have been studied by means of acrylamide gel disc electrophoresis. A maximum of eighteen bands were observed in the male of the 6th instar larva and the female of the late pupa. In general, the protein bands increased with the growth of larva, decreased at the prepupal stages, and at the pupal stages they increased again. The possible significance of these observations has been discussed. In addition, comparisons have been made of the protein patterns between the males and females in the developmental stages.

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Chemopreventive Effect of Vegetable or Fruit Extract Against Total Diesel Exhaust Particle Extract in NIH/3T3 Cells Using Alkaline Single Cell Gel Electrophoresis (총 디젤분진의 DNA 손상작용과 야채 및 과일추출물의 보호효과)

  • Heo Chan;Kim Nam-Yee;Heo Moon-Young
    • Environmental Analysis Health and Toxicology
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    • v.21 no.2 s.53
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    • pp.127-138
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    • 2006
  • In urban areas, diesel exhaust particles (DEP) are probably a major component of particulate matters, especially in Korea where drive many diesel vehicles. The aim of this study was to investigate genotoxic effects of DEP using single ceil gel electrophoresis. In order to evaluate the mechanisms of DEP genotoxicity, the rat microsome mediated and DNA repair enzyme treated comet assays together with conventional comet assay were performed. Total diesel particles (DEPT) was collected without site fractionation from diesel engine bus and dichloromethane extract was obtained. The organic extract of DEPT revealed DNA damage itself in NIH/3T3 cells. The level of DNA breaks plus oxidative DNA lesions and microsome mediated DNA damage was assessed by modified single cell gel eletrophoresis. DEPT was able to induce oxidative DNA damage as well as microsome mediated DNA damage. Vitamin C as an model antioxidant reduced DNA damage in endonuclase III treated comet assay. One of flavonoid, galangin as a CYP1A1 inhibitor. reduced DNA damage in the presence of S-9 mixture. $DEP_T$ is the sources of oxidative stress, but antioxidants can significantly reduce oxidative DNA dmage. And $DEP_T$ may contain indirect mutagens which can be inhibited by CYP1A1 inhibitors. The ethanol extracts of the mixed vegetables (BV) or the mixed fruits (BF) were evaluated for their in vitro antigenotoxic effects. BV and BF showed potent Inhibitory effects against DEPT induced DNA damage with oxidative DNA lesions and in the prescence of S-9 mixture. These results indicate that BV and BF could prevent cellular DNA damage by inhibiting oxidative stress and suppressing cytochrome P4501A1 in cell culture.

Proteomic Analysis of Bovine Pregnancy-specific Serum Proteins by 2D Fluorescence Difference Gel Electrophoresis

  • Lee, Jae Eun;Lee, Jae Young;Kim, Hong Rye;Shin, Hyun Young;Lin, Tao;Jin, Dong Il
    • Asian-Australasian Journal of Animal Sciences
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    • v.28 no.6
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    • pp.788-795
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    • 2015
  • Two dimensional-fluorescence difference gel electrophoresis (2D DIGE) is an emerging technique for comparative proteomics, which improves the reproducibility and reliability of differential protein expression analysis between samples. The purpose of this study was to investigate bovine pregnancy-specific proteins in the proteome between bovine pregnant and non-pregnant serum using DIGE technique. Serums of 2 pregnant Holstein dairy cattle at day 21 after artificial insemination and those of 2 non-pregnant were used in this study. The pre-electrophoretic labeling of pregnant and non-pregnant serum proteins were mixed with Cy3 and Cy5 fluorescent dyes, respectively, and an internal standard was labeled with Cy2. Labeled proteins with Cy2, Cy3, and Cy5 were separated together in a single gel, and then were detected by fluorescence image analyzer. The 2D DIGE method using fluorescence CyDye DIGE flour had higher sensitivity than conventional 2D gel electrophoresis, and showed reproducible results. Approximately 1,500 protein spots were detected by 2D DIGE. Several proteins showed a more than 1.5-fold up and down regulation between non-pregnant and pregnant serum proteins. The differentially expressed proteins were identified by MALDI-TOF mass spectrometer. A total 16 protein spots were detected to regulate differentially in the pregnant serum, among which 7 spots were up-regulated proteins such as conglutinin precursor, modified bovine fibrinogen and IgG1, and 6 spots were down-regulated proteins such as hemoglobin, complement component 3, bovine fibrinogen and IgG2a three spots were not identified. The identified proteins demonstrate that early pregnant bovine serum may have several pregnancy-specific proteins, and these could be a valuable information for the development of pregnancy-diagnostic markers in early pregnancy bovine serum.

Microbial Community Structure of Korean Cabbage Kimchi and Ingredients with Denaturing Gradient Gel Electrophoresis

  • Hong, Sung Wook;Choi, Yun-Jeong;Lee, Hae-Won;Yang, Ji-Hee;Lee, Mi-Ai
    • Journal of Microbiology and Biotechnology
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    • v.26 no.6
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    • pp.1057-1062
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    • 2016
  • Kimchi is a traditional Korean fermented vegetable food, the production of which involves brining of Korean cabbage, blending with various other ingredients (red pepper powder, garlic, ginger, salt-pickled seafood, etc.), and fermentation. Recently, kimchi has also become popular in the Western world because of its unique taste and beneficial properties such as antioxidant and antimutagenic activities, which are derived from the various raw materials and secondary metabolites of the fermentative microorganisms used during production. Despite these useful activities, analysis of the microbial community present in kimchi has received relatively little attention. The objective of this study was to evaluate the bacterial community structure from the raw materials, additives, and final kimchi product using the culture-independent method. Specifically, polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) was used to analyze the 16S rRNA partial sequences of the microflora. One primer set for bacteria, 341FGC-518R, reliably produced amplicons from kimchi and its raw materials, and these bands were clearly separated on a 35-65% denaturing gradient gel. Overall, 117 16S rRNA fragments were identified by PCR-DGGE analysis. Pediococcus pentosaceus, Leuconostoc citreum, Leuconostoc gelidum, and Leuconostoc mesenteroides were the dominant bacteria in kimchi. The other strains identified were Tetragenococcus, Pseudomonas, Weissella, and uncultured bacterium. Comprehensive analysis of these microorganisms could provide a more detailed understanding of the biologically active components of kimchi and help improve its quality. PCR-DGGE analysis can be successfully applied to a fermented food to detect unculturable or other species.

Studies on Serum Albumin Types in Goats by Starch Gel Electrophoresis (전분(澱粉) Gel 전기영동법(電氣泳動法)에 의한 염소의 혈청(血淸) Albumin 형(型)에 관한 연구(硏究))

  • Lee, Jai Hong
    • Korean Journal of Veterinary Research
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    • v.15 no.1
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    • pp.19-22
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    • 1975
  • The serum albumin phenotypes and the gene frquencies of 100 Korean native goats were examined by starch gel electrohporesis. The results obtained were as follows: 1. The albumin phenotypes were classified as Alb AA, Alb AB, and Alb BB and the frequencies of appearence were 10, 12 and 78%, respectively. 2. The genetic factors of serum albumin were observed as Alb A and Alb B and the rates of gene frequencies were 16 and 84%, respectively.

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Purification of Glucose Isomerase from Arthrobacter sp. L-3 (Arthrobacter sp. L-3이 생성하는 Glucose Isomerase의 정제)

  • 방병호;남궁석
    • The Korean Journal of Food And Nutrition
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    • v.4 no.2
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    • pp.155-160
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    • 1991
  • Authrobacter sp. L-3이 생성하는 glucose isomerase를 DEAE-cellulose column chromatography법으로 2단계 NaCl농도 구배로 용출함으로서 순수분리하였다. 이것이 SDS-acrylamide gel electrophoresis상에서 단일띠를 보임으로서 매우 잘 분리되었음을 알 수 있었다. Glucose isomerase의 Km값과 Vmax값이 각각 0.175M, 0.29로 얻어졌다. 한편, SDS-acrylamide gel electrophoresis와 Sephadex-G10050에 의한 gel filtration으로부터 분자량이 각각 42,000과 180,000으로 얻어져, 이 효소는 분자량이 42,500인 4개의 subunit로 구성되었음을 알 수 있었다.

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Changes of Protein During Growth of Soybean Sprout (Gel filtration에 의한 콩나물 제조중(製造中) 단백질(蛋白質)의 변화조사(變化調査))

  • Yang, Cha-Bum;Park, Sang-Ki;Yoon, Suk-Kwon
    • Korean Journal of Food Science and Technology
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    • v.16 no.4
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    • pp.472-474
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    • 1984
  • Gel filtration of water soluble protein of soybean and cotyledon of sprout at various growth stages by using Sephadex G-200 showed 5 fractions (A,B,C,D and E). According to gel filteration and disc gel electrophoresis, fraction B,C and D were identified as 11S,7S and 2S, respectively. Fraction A was turbid substance and fraction E showed 1 peak. During growth of sprout 7S decreased firstly, 2S secondly and 11S lastly in cotyledon. Fraction A increased until 6th day and decreased thereafter while E increased throughout the growth. In axis only two fractions (11S+7S and E) were showed, and 11S+7S fraction was little changed and fraction E increased slightly with the growth.

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