• Title/Summary/Keyword: SDS-PAGE pattern

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Optimum pH Condition of Defatted Rice Protein Extraction by Alkaline Method (알칼리 추출법에 의한 탈지 미강 단백질 추출의 최적 pH 조건)

  • Kim, Won;Jung, So-Young;Hong, Kwang-Won
    • Food Engineering Progress
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    • v.15 no.2
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    • pp.143-147
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    • 2011
  • For efficient extraction of protein from defatted rice bran, the 5 ranges of extraction pH (8, 9, 10, 11 and 12) and the 3 ranges of isoelectric precipitation pH (2, 4 and 6) were used. The protein content, browning reaction, the electrophoresis pattern and the recovery yield of soluble protein at each pH range were compared each other. The recovery yield of soluble protein increased in proportion to extraction pH, but at the same time, browning reaction became more conspicuous. The most amount of protein was recovered at the precipitation pH of 4. The SDS-PAGE patterns of the extracted proteins showed no significant correlations between pH and the protein content, but the highly alkaline condition was more advantageous to extract protein less than 35 kDa. In each pH range, the recovery yield of soluble protein averagely reached 32.5% on the basis of extraction. In result, it was found that combination of extraction pH 10 and precipitation pH 4, which resulted in 37.65% of recovery yield and low level of browning reaction, was the optimum condition for the extraction of protein from defatted rice bran.

Effect of the paper acidity on the cellulolytic activity of fungi (종이의 산성화가 미생물의 분해능에 미치는 영향)

  • Han, Sung-Hee;Lee, Kyu-Shik;Chung, Young-Jae;Lee, Hye-Yun
    • 보존과학연구
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    • s.19
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    • pp.3-22
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    • 1998
  • The effect of pH on degradation of paper by some fungi, which able to degrade cellulose, was investigated. Trichoderma koningii, Aspergillus nigerand Penicillium nigulosum were cultured at $28^{\circ}C$ for 16 days in the selective medium (PH3, PH4, PH5, PH6, PH7, PH8, PH9, PH10, PHC) containing paper as substrate. Each paper was pretreated with each pH buffer (pH 3∼pH 10, D.W.)prior to addition to the selective medium. Enzyme activities in the each culture medium were measured spectroph to metrically using C.M.C., Avicel, PNPG as the substrates for endoglucanase, exoglucanase and $\beta$-glucosidase, respectively. In all experimental fungi, the enzyme activities of PH3 and PH9 medium were usually much higher than those of other experimental groups. However in the PH6medium, enzyme activity was lower than other groups. To analyze the concentration and pattern of protein in the each culture medium, the medium was concentrated by lyophilization. The protein concentration of PH3 and PH9 medium were relatively high (T.koningii; 6.31mg, 6,19mg, A.niger; 1.62mg, 1.96mg, P.nigulosum;2.50mg, 2.73mg, respectively), but that of PH6 was relatively low. The protein pattern of each medium was analyzed by using SDS-PAGE and VDS Image Master Analysis Program. The concentrations of bands in the each lane were usually high at lane2 (PH3) and lane8 (PH9) and low at lane5 (PH6). Therefore, the incresed cellulolytic activity of fungus against acidified paper could be result of structural change and deterioration of paper caused by being acidified.

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Characterization of Cucumver mosaic virus Isolated from Hydrangea macrophylla for. otaksa (Sieb. et Zucc) Wils. (수국에서 분리한 Cucumber mosaic virus의 특성)

  • 방주희;박선정;이금희;최장경;이상용
    • Research in Plant Disease
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    • v.7 no.1
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    • pp.1-7
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    • 2001
  • An isolate of Cucumber mosaic cucumovirus(CMV) was isolated from Hydrangea macrophylla for. otaksa(Sieb. et Zucc. ) Wils. showing mosaic symptoms, and designated as Hm-CMV. Hm-CMV was characterized by the tests of host range, physical properties, serological properties, RNA and coat protein compositions, and reverse transcription and polymerase chain reaction (RT-PCR) analysis. Twelve species in 4 families were used in the host range test of Hm-CMV and could be differentiated from Y-CMV used as a control CMV by the ringspot and line pattern on inoculated leaves of several tobacco plants. Thevirus produced local lesions on inoculated leaves of Chenopodium amarticolor, C. quinoa and Vigna unguiculata. The physical properties of the virus were as follows; thermal inactivation point(TIP) was 60$\^{C}$, dilution end point (DEP) was 10$\^$-3/, and longevity in vitro (LIP) was 3∼4 days. Hm-CMV was serologically identical to Y-CMV. SDS-polyaciylamide gel electrophoresis(SDS-PAGE) showed one major protein band of about 28 kDa. In RNA or dsRNA analysis, Hm-CMV consisted of four RNA or dsRNA species, but satellite RNA was not detected. In RT-PCR using CMV-common primer and CMV subgroup I-specific primer, bothe amplified expected size of about 490 bp and 200 bp DNA fragments from Hm-CMV, respectively. Restriction enzyme analysis of the 490 bp RT-PCR products using EcoR I and Msp I showed that Hm-CMV belonged to CMV subgroup I. However, Hm-CMV could be differentiated from other CMV subgroup I isolates by RNA fingerprinting by arbitrarily primed polymerase chain reaction (RAP-PCR).

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A Comparison of Protein Characteristics of Korean and Imported Wheat Varieties (한국산 소맥과 수입 소맥의 단백질 특성 비교)

  • Koh, Bong-Kyung
    • Korean Journal of Food Science and Technology
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    • v.31 no.3
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    • pp.586-592
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    • 1999
  • The objective of this study was to investigate the fundamental characteristics of proteins effecting on Korean wheat flour quality. Mixing properties and protein contents of Eunpa, Allgreu, and Woori showed soft wheat characteristics. Tapdong had strong dough stability and high protein content like hard wheat flour. However, even though Greu had a high protein content, the dough strength was very weak like soft wheat flour. No major difference was observed in protein solubility with water, 0.1N NaCl, 70% ethyl alcohol, 0.1 N acetic acid solution and in sulfur and disulfide contents between Korean wheat flours and imported flours. However, DNS (Dark Norther Spring) had more contents of acid insoluble and SDS insoluble proteins compared with Korean wheat varieties. PAGE pattern of Tapdong was very similar to imported wheat flours at the high molecular weight regions and characteristic high molecular weight glutenin subunits were observed over 116 kD protein molecular weight region from 4 Korean wheat flours such as Eunpa, Woori, Allgreu, and Greu.

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Genetic characteristics of the novel insect pathogenic Bacillus thuringiensis subsp. aizawai strain (새로운 곤충병원성 Bacillus thuringiensis subsp. aizawai 균주의 유전학적 특성)

  • Seo, Mi Ja;Youn, Young Nam;Yu, Yong Man;Kim, Ki Su
    • Korean Journal of Agricultural Science
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    • v.41 no.4
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    • pp.351-359
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    • 2014
  • For identifying the plasmid DNA coding cry gene of Bacillus thuringiensis subsp. aizawai KB098 with high insecticidal activity against Spodoptera exigua, mutant isolates with no crystal protein were produced by $42^{\circ}C$ incubation condition and then mutant plasmid DNA band patterns were compared with those of KB098. KB098 isolates had 4 cry genes, cry1Aa, cry1Ab, cry1C, cry1D, and also had been found seven plasmid DNA. Though the SDS-PAGE experiment, it was confirmed that mutant didn't produce 130~145kDa protein band involved in bipyramidal shape crystal. Also, five mutant isolates had no cry genes coding plasmid DNA in PCR. In result of comparison the plasmid DNA of KB098 and 5 mutant isolates, only 1 plasmid DNA band was left out in mutant plasmid DNA pattern, so that the missing band was extracted from the gel. The missing(disappeared) plasmid DNA was the largest molecular size among the 7 plasmid DNA of KB098 and it was also confirmed this plasmid DNA had all 4 cry genes through PCR.

복제 소 태반과 IVF 소 태반의 protein pattern 분석

  • 김홍래;강재구;윤종택;성한우;조민래;박창식;진동일
    • Proceedings of the Korean Society of Developmental Biology Conference
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    • 2003.10a
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    • pp.90-90
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    • 2003
  • 체세포 핵이식에 의한 복제기술은 매우 낮은 성공률 나타내고 있어 실용화에 지장을 초래하고 있다. 이것은 후생적인 유전현상인 reprogramming이 불완전하게 이루어지기 때문인 것으로 추측되어지고 있다(Reik et al., Theriogenology 2003, 59: 21-32; Han et al, Theriogenology 2003, 59: 33-44). 체세포 핵이식 후에 태아사망의 원인이 태반의 비정상적인 기능과 관계가 있는 것으로 추정되는데 복제시 태아사망의 원인을 찾기 위해 본 연구를 시행하였다. 한우에서 체세포 복제 후 임신 말기에 태아가 사망한 태반조직 3개와 IVF 수정란 이식 후 동일한 시기에 제왕절개술을 실시한 태반조직 2개를 실험에 이용하였다. 태반 protein을 Two-Dimensional electrophoresis와 Mass spectrometer를 이용하여 분석 비교하였다. IPG-system을 이용하여 pH 4~7, pH 6~9에서 1차 전기영동을 한 후, 8~l6%의 SDS-PAGE gel에 2차 전기영동을 실시하였고 G-250 Coomassie로 염색하였다. gel 이미지는 Malanie III program을 이용하여 분석하였다. 전체 gel에서 약 1800개의 구분 가능한 protein spot이 나타났다. pH 4~7 범위에서 양적으로 차이나는 것 15개 중 복제한우 태반에서 증가되는 protein spot 5개와 감소하는 protein spot 10개를 골라 protein identification을 실시하였다. MALDI-TOF-MS를 이용하여 동정한 결과 phosphatidylinositol transfer protein-$\alpha$와 interleukin-18 등의 protein이 복제태반에서 발현이 증가되었고, 복제한우에서 발현이 감소되는 것으로는 vimentin, Rho-GDI-$\beta$, TRAST $\beta$-chain, ovarian sterol carrier protein 2, triosephosphate isomerase, tropemyosin beta chain, Aldose reductase 등으로 나타났다. 이러한 protein들은 inositol 지질 신호전달과 면역시스템, 세포분열, 산소 운반, steroidogenic 세포에서의 콜레스테롤 이동, 촉매 작용, 대사 작용 등에 중요한 역할을 하는 것으로 알려져 체세포 복제에 의한 태아사망 원인은 태반에서 이러한 protein들의 비정상적인 발현에 기인된 것으로 추정된다.

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Characterization of Vibrio vulnificus Isolated from Domestic Coastal Area (국내 연안에서 분리한 Vibrio vulnificus의 특성)

  • Park Geun-Tae;Park Min Jung;Jung Cho-Rok;Song Choon-Bok;Lee Jehee;Yeo In-Kyu;Jeon You Jin;Heo Moon-Soo
    • Journal of Life Science
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    • v.14 no.6 s.67
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    • pp.986-990
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    • 2004
  • Five strains of Vibrio vulnificus, which cause serious septicemia in human worldwidely, was isolated from marine environments of Korea costal area from May to July of 2002. The isolated strains were identified by API 20E kit and partial 16S rDNA sequence analysis. 16S rDNA sequence of the isolates showed $99\%$ similarity with V. vulnificus ATCC 27562. The proteins of V. vulnificus isolates were examined by analyzing patterns of the cell lysates and outer membrane proteins (OMP). The OMP separated from cell lysates showed the common protein band. Therefore OMP profiles might be useful for the identification of V. vulnificus sp.

Characteristics of ustilago maydis virus of SH14 killer strain isolated in Korea

  • Hwang, Seon-Hee;Jung, Cheong-Hwan;Yie, Se-Won
    • Journal of Microbiology
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    • v.33 no.2
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    • pp.154-159
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    • 1995
  • SH-14, a novel killer strain of Ustilago maydis was isolated in Korea. It has been reported in other papers that the toxin specificity and double-stranded RNA pattern of SH-14 strain were different from other laboratory strains. In this paper, we analyzed the biochemical characteristics of U. maydis SH-14 virus. Three distinctive peaks were isolated from CsCl density gradient, designated as top (T), intermediate (I) and bottom (B) components. We found that the densities of each components, 1.285, 1.408 g/cm$\^$3/, respectively, are very similar to those of other strains. As previously reported by the analysis of dsRNA in each component, the dsRNA segments are separately encapsidated. Capsid protein of SH-14 virus consists of two proteins about 70 Kd shown by SDS-PAGE analysis. Electron microscopic examination of the virus particles revealed that UmV particles are very similar in size and morphology to all isolates as well as all lab-strains. In order to test immunological cross reactivity of UmV, werstern bolt analysis was carriedout with antiserum against A8 virus. All capsid protein had positive reaction against A8 antibody which indicated that UmV are immunologically cross-reactive with all isolates from Korea. The results presented in this paper may show that UmV isolated from SH-14 strain has very similar biochemical characteristics to those of other UmV. However, the difference in the toxin specificity and the molecular weight of toxin protein from the SH-14 strain has us to conclude that U. maydis SH-14 strain is a new killer type.

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Characterization of Antimicrobial Substance Produced by Lactobacillus paraplantarum KNUC25 Isolated from Kimchi (김치로부터 분리된 Lactobacillus paraplantarum KNUC25가 만드는 항균 물질의 특성)

  • Kim, Ma-Rie;Lee, Su-Jin;Seul, Keyung-Jo;Park, Yu-Mi;Ghim, Sa-Youl
    • Microbiology and Biotechnology Letters
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    • v.37 no.1
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    • pp.24-32
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    • 2009
  • The KNUC25 strain isolated from over-fermented whole Chinese cabbage kimchi was examined for its physiological characteristics using API 50 CHL system assay and identified as Lactobacillus paraplantarum by analysis of whole-cell protein SDS-PAGE pattern assay and similarity of 16S rDNA sequence. L. paraplantarum KNUC25 had a broad antimicrobial activity spectrum from Gram positive to Gram negative bacteria. Scanning electron micrograph analysis showed that KNUC25 might attack to cell surface of indicator cells and destruction can lead to inhibition of the cell growth. The antimicrobial substance of the KNUC25 strain was stable to various degrading enzymes and at high temperature and not a plasmid-born matter. Resistance to proteolytic enzymes showed that an antimicrobial activity of KNUC25 might not be caused by proteinous substance. Maximum production of antimicrobial substance was the exponential growth phase at $30^{\circ}C$.

Effects of cultural conditions on growth of Micrococcus sp. and casein hydrolysis : (II) -Studies on patterns of casein hydrolysis with time during culture- (Micrococcus sp.의 생육 및 casein 분해에 미치는 배양조건의 영향 : (II) -배양시간에 따른 casein 분해 형태에 관한 연구-)

  • Lee, Si-Kyung;Pec, Un-Hua;Joo, Hyun-Kyu
    • Applied Biological Chemistry
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    • v.35 no.6
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    • pp.479-484
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    • 1992
  • This study was undertaken to determine the effects of cultural conditions on cell growth and casein hydrolysis for cell production in order to add Micrococcus sp. LL3 as a potential agent for industrial application with aim of shortening ripening period and improving flavor. Optimum temperature for cell growth and caseinolysis was $30^{\circ}C$ and $37{\circ}C$, respectively, and optimum pH was 7.0. The enzyme remained stable up to $50^{\circ}C$. Hydrolysis patterns of casein were also observed on SDS-PAGE. Both ${\alpha}-casein$ and ${\beta}-casein$ were totally hydrolysed by enzymes from Micrococcus sp. LL3 during culture. A preferential attack on ${\beta}-casein$ was observed. Production of aminopeptidase which cleaved polypeptides was the highest in early stationary phase during cell growth.

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