• Title/Summary/Keyword: SDS-PAGE pattern

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Structural Characterization of Hordeum vulgare L. Chloroplast by Ozone

  • Chung, Hwa-Sook;Lim, Young-Jin;Park, Kang-Eun
    • Environmental Sciences Bulletin of The Korean Environmental Sciences Society
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    • v.4 no.2
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    • pp.85-94
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    • 2000
  • The effects of ozone on chloroplast development in barley seedlings during greening was investigated based on ultrastructural changes in the chloroplasts and band pattern changes in the chloroplast thylakoid membrane proteins. In this analysis of the chloroplast thylakoid membrane thylakoid protein band pattern by SDS-PAGE, none of the 24-hour greening bands included were clearer than the control. This means that the ozone treatment produced a dealy in chloroplast development and decreased the amount of thylakoid membrane proteins. LHC II chloroplast band of developing barley seedlings treated with 0.5 and 1.0 ppm ozone during the last 4 hours of the 24-hour greening period was weaker than the other bands. This result indicates that ozone affects the LHC II protein complex of the chloroplast thylakoid membrane. When investigating the ultastructural changes in ozone-treated chloroplast, the main site affected by 0.5 ppm ozone was the chloroplast grana, thereby explaining the delayed chloroplast development during the early phase of greening. In addition, there was also a structural change in the stromal grana of the ozone treated chloroplast during the middle phase of greening. The effects of ozone on the chloroplast of barley seedlings during the last phase of 48-hour greening were more functionally inhibiting than structural changes.

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Large-Scale Culture of Hepatitis A Virus in Human Diploid MRC-5 Cells and Partial Purification of the Viral Antigen for Use as a Vaccine

  • Kim, Hyun-Seok;Chung, Yong-Ju;Jeon, Yeong-Joong;Lee, Sung-Hee
    • Journal of Microbiology and Biotechnology
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    • v.9 no.4
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    • pp.386-392
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    • 1999
  • A large-scale culture of hepatitis A virus in human diploid MRC-5 cells was conducted. In a roller bottle culture, the virus was grown to a maximum titer in 3 weeks after infection. Over 95% of the cell-associated virus was excreted after culturing the infected cells in suspension media without fetal bovine serum for 3 days. The cultured virus was inactivated with formalin, concentrated by ultrafiltration, and partially purified by ultracentrifugation in a non-ionic gradient medium of Renocal. Two separate peak fractions showing high anti-HAY ELISA titer were pooled and about 40% of HAV antigen was recovered by this purification procedure. Of the partially purified vaccine, the protein pattern in SDS-PAGE and immunogenicity in mice were compared with a commercial HAV vaccine. In SDS-PAGE, the purified vaccine in this study and the commercial vaccine showed almost the same protein pattern. The seroconversion rate of the purified vaccine in mice was not different from that of the commercial vaccine. Therefore, we could prepare a good grade of HAV vaccine by a simple purification procedure although the purification itself was not completed.

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Effects of gamma irradiation on the functional and structural characteristics of soy protein isolates (방사선 조사가 분리대두단백의 기능적, 구조적 특성에 미치는 영향)

  • 오경남;이숙영
    • Korean journal of food and cookery science
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    • v.20 no.3
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    • pp.256-264
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    • 2004
  • The effects of irradiation on the functional and structural characteristics of soy protein isolates were studied. Soymilk was irradiated at 1, 5, and l0kGy, after which soy protein isolates were prepared. The functional properties of soy protein isolates were examined including solubility, emulsion capacity and stability, foam capacity and stability, structural properties as represented by SDS-PAGE pattern, and secondary and tertiary structures. The solubility and emulsion capacity were increased by radiation treatment at 1kGy however the values were adversely affected again as dosage was increased above 5kGy. As irradiation dosage increased, an increase of foaming capacity at 1kGy and a decreasing turnover afterwards were also noted in foaming capacity, although the differences were not statistically significant. The SDS-PAGE pattern showed fragmentation and aggregation of protein molecules as affected by irradiation in proportion to the dosage increase. The results of CD and fluorescence spectroscopy revealed increased aperiodic structure contents with the dosage increase. It was assumed that irradiation dosagefrom 5 to l0kGy could initiate minimal denaturation of protein in various foods compared to general heat treatment.

Changes of SDS-PAGE Pattern of Pork Myofibrillar Proteins Induced by Electron Beam Irradiation

  • Whang Key;Jeong, Dong-Kwan;Kim, Hyuk-Il
    • Preventive Nutrition and Food Science
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    • v.10 no.4
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    • pp.378-381
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    • 2005
  • Actin and myosin solutions and fresh ground pork were irradiated with the electron beam (e-beam) at a dose of 0, 1.5, 3.0, 5.0 and 10 kGy. The changes in SDS-PAGE pattern of 2 proteins and the salt-soluble proteins extracted from ground pork after e-beam irradiation were monitored. When the myosin solution was irradiated with e-beam, myosin was degraded completely. Complete myosin degradations were observed even with the lowest dose (1.5 kGy) of e-beam treatment. Actin was degraded with the irradiation, but to a less extent than myosin was. The degradation of actin increased as the e-beam treatment increased from 1.5 to 10.0 kGy. Among the salt-soluble proteins extracted from ground pork, myosin was degraded gradually when the e-beam dose increased from 1.5 up to 10.0 kGy. Similar gradual increase in the degradation of actin also occurred with the increase of irradiation. Increases of 2 low molecular weight compounds (<29 kDa) were observed when the irradiation dose increased from 1.5 to 10.0 kGy. These 2 molecules are thought to be the breakdown products produced from the degradation of major salt-soluble proteins, myosin and actin. The salt-soluble protein content of ground pork did not change with the e-beam irradiation.

Modification of Functional Properties of Soy Protein Isolate by Proteolytic Enzymes (단백분해효소에 의한 대두단백의 기능적 특성변화)

  • Cha, Myeong-Hwa;Yoon, Sun
    • Korean Journal of Food Science and Technology
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    • v.25 no.1
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    • pp.39-45
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    • 1993
  • The effects of enzymatic modification with pepsin and actinidin was studied on molecular weight distributions and functional properties of hydrolysates from soy protein isolate (SPI) differing in degree of hydrolysis. The hydrolyzed SPI by pepsin showed 41.5% degree of hydrolysis after 5 min, and maximum hydrolysis was obtained after 2 hours. Actinidin hydrolyzed SPI 26.71% degree after 1 hour. On SDS-PAGE, native SPI showed 9 distinguishable bands on SDS-PAGE gel. Pepsin treated SPI showed one broad band in the lower part of gel. This band was shifted further to the bottom of the gel and became faint as hydrolysis time increased. While actinidin treated SPI showed different SDS-PAGE pattern from pepsin. However PAGE patterns were similar with pepsin and actinidin treated groups. With pepsin treatment, solubility of SPI distinctively increased around isoelectric point(pI). Emulsifying activity (EA) and emulsifying stability (ES) showed marked increase over pH range of $3.0{\sim}8.0$. 5 min modified group had most excellent foam expansion (FE). Foam stability (FS) was increased as pepsin treatment time increased at pI. With actinidin treatment, solubility was increased. 60 min modified SPI had the most effective EA at pH 4.5. However ES was not effected by actinidin treatment. 5 min modified group was most effect in FE. FS was higher at alkaline pH.

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Changes of Protein Pattern of Mungbean Seeds, Phaseolus aureus During Germination (녹두 발아중 단백질 전기영동 패턴의 변화)

  • Park, D.Y.;Cho, S.J.;Shin, Y.C.
    • Korean Journal of Food Science and Technology
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    • v.18 no.2
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    • pp.163-167
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    • 1986
  • During the germination of mungbean seeds, the changes of water contents, total and soluble proteins, and electrophoretic pattern of the soluble proteins were examined. The moisture content of a dry mungbean was 12.7%, which was greatly increased after the soaking. Along to the germination period, the moisture contentof the mungbean sprouts was gradually increased up to 90.7%. The contents of total and soluble proteins were sharply decreased after the soaking of the mungbean and decreased gradually during the germination. PAGE of the soluble proteins showed two broad bands and three sharp bands. During the germination, two broad bands were weadened but other bands were relatively stable. SDS-PAGE showed 19 discrete bands and during the germination, the most of the bands were thinned or disappeared. But some of the protein bands were stable until the end of germination.

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Identification of Different Species and Dultivars of Brassica by SDS-PAGE, Isozyme and Molecular Marker

  • Mukhlesur Rahman Md.;Hirata Yutaka
    • Journal of Plant Biotechnology
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    • v.7 no.1
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    • pp.27-35
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    • 2005
  • Eighty-five different cultivars of Brassica rapa, B. juncea, B. nap us, B. carinata, B. oleracea and hexaploid Brassica collected from Bangladesh, Japan, China and Denmark were analyzed by SDS-PAGE for seed and leaf protein variations, using esterase, acid phosphatase and peroxidase isozyme analysis. Ten polymorphic bands were identified from seed protein however no identifiable polymorphic band was found in the leaf protein. Polymorphic markers clearly distinguished the different Brassica species as well as yellow sarson (YS) and brown seeded (BS) cultivars of B. rapa. The $F_1$ cross between YS and brown seeded cultivars showed the existance of all poly-morphic bands of the respective parents. The Bangla-deshi and Japanese cultivars of B. rapa differed in the amount of seed protein. In the case of isozyme analysis, esterase showed the highest number of polymorphic bands (13) followed by acid phosphatase (9) and peroxidase (5). These polymorphic markers were very effec-tive for classification of all the species studied in this experiment. In parentage tests using isozymes, the hybridity of intra-and-interspecific crosses of almost all the seedlings could be identified from their respective cross combinations. Esterase polymorphism showed a clear differentiation between YS and BS types of B. rapa. In addition, two esterase polymorphic markers were iden ified to differentiate some cultivars of B. juncea. Segregation patterns in these two esterase bands showed a simple Mendelian monohybrid ratio of 3:1 in $F_2$, 1:1 in test cross and 1:0 in back cross progenies. No polymorphic band was identified to distinguish different cultivars of the same species by acid phosphatase or peroxidase. Polymerase Chain Reaction (PCR) was carried out with seed coat color specific marker of B. juncea. The yellow seeded cultivars produced a strong band at 0.5 kb and weak band 1.2 kb. In the addition of these two specific bands, Japanese yellow-seeded cultivars expressed two more weak bands at 1.0 kb and 1.1 kb. Where the brown seeded cultivars generated a single strong band at 1.1 kb. In segregating population, the yellow seed coat color marker segregated at a ratio 15 (brown) : 1 (yellow), indicating the digenic inheritance pattern of the trait.

A Study on the Pattern of Proteins on Electrophoresis in the Porcine Follicular Fluid During Atresia (폐쇄에 따른 돼지 여포액내 단백질의 전기영동 양상에 관한 연구)

  • Kim, Jong-Heup;Yoon, Yong-Dal;Kim, Moon-Kyoo
    • Clinical and Experimental Reproductive Medicine
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    • v.13 no.1
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    • pp.39-51
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    • 1986
  • This experiment has been done to evaluate the relationship between the follicular atresia and the protein patterns on electrophoreais of the follicular fluids in porcine ovary. The protein concentration of the follicular fluids was lower than that of serum, and gradually decreased as the follicle siae became larger. The number of protein bands of follicular fluid on electrophoresis was less than that of serum, and gradually increased as follicle size became larger. Three specific bands were detected on disc PAGE and one band(M W. 75,000) on SDS PAGE in the follicular fluids, while not in serum. One band (A) at ${\beta}$-globulin region on disc PAGE became heavier, as follicles became atretic. Two bands less than(M. W. 20,000) were detected only in the large follicular fluid. Another band(M. W. 43,000) was not detected in necrotic group, whereas all other groups showed it. It could be concluded that the component and composition of the proteins follicular fluids changes according to the follicular size during atresia. Therefore detection of the changing pattern of proteins in the follicular fluid can be used as a basic criterion for the identification of follicular atretic stage.

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Electrophoretic Patterns of Hemolymph Proteins of Varieties with Long and Short Life Span in the Silkworm Bombyx mori L. (${\cdot}$단명 누에 품종의 체액단백질의 전기영동상)

  • Kang, Pil-Don;Yoon, Hyung-Joo;Ryu, Kang-Sun;Sohn, Bong-Hee;Sohn, Hung-Dae
    • Journal of Sericultural and Entomological Science
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    • v.41 no.1
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    • pp.1-8
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    • 1999
  • Hemolymph protein patterns of silkworms in terms of short and long life span were analyzed by native- and SDS-polyacrylamide gel electrophoresis (PAGE) with the developmental stages. From the native-PAGE patterns of silkworm major hemolymph proteins there were varietal differences on the first day of the pupal stage and were classified into there groups MHP-a, b and c SA 10, JAM109 and J037 were grouped into MHP-Ⅰ, Hangang, Chungmun and Daizo(sdi) into MHP-Ⅱ and NTZN, Sulak, Qoichuk and PR varieties into MHP-Ⅲ group. It was found that the MHP in each group revealed similar patterns and changes with development of pupal stage. In the first day of adult, MHP-c was clearly detected in both female and male of the Daizo(sdi), but not in the J037, indicating that there was significantly varietal differences in electrophoretical protein pattern. In addition, the results of protein pattern of hemolymph by SDS-PAGE showed also varietal differences in the concentration of hemolymph protein.

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Biochemical Properties of Seed Lectin from Korean Soybean Cultivars Developed for Soy Source (한국산 장류콩 종자 렉틴의 생화학적 특성)

  • Wang, Yushan;Roh, Kwang-Soo
    • KSBB Journal
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    • v.24 no.2
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    • pp.170-176
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    • 2009
  • Lectin was finally isolated on Sephadex G-100 from Korean soybean cultivars developed for soy source and investigated its some biochemical properties. Native PAGE pattern of this lectin revealed a molecular weight of 108 kDa as tetramer. The molecular weight of this lectin isolated as double protein band by SDS-PAGE was calculated to be 32 and 22 kDa from the relative mobilities compared with those of the standard proteins. Among the tested red blood cell, the isolated lectin agglutinated rabbit red blood cell treated with trypsin, but did not agglutinated human red blood cells (A, B, AB, O), rat, and untreated rabbit red blood cell. The optimal temperature and thermal stability of isolated lectin was at 20-$50^{\circ}C$ and 10-$60^{\circ}C$, respectively. This lectin was stable at 7.2, and showed complete loss in its activity below pH 6.2 and above pH 8.0.