• Title/Summary/Keyword: isoelectrofocusing

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Purification of Trichoderma viride Cellobiohydrolase by Immunoaffinity Chromatography (면역친화 크로마토그라피에 의한 Tricholderma viride의 Cellobiohydrolase 분리)

  • 오태광
    • Microbiology and Biotechnology Letters
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    • v.18 no.4
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    • pp.390-393
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    • 1990
  • A cellobiohydrolase was purified from the culture broth of Trichoderma uiride by using immunoaffinity chromatography. A single protein band in polyacylamide gel electrophoresis and isoelectrofocusing after immunoaffinity purification corresponded to cellobiohydrolase activity. A immunoaffinity purified cellobiohydrolase is more effective in the hydrolysis of highly crystalline cellulose than amorphous cellulose.

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Genetic analysis and characteristics in the crescent-egg mutant, cre, of Bombyx mori. (신돌연변이잠 cre(반월형란)의 유전자 연관분석과 유전형질)

  • 홍선미;노시갑
    • Journal of Sericultural and Entomological Science
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    • v.43 no.2
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    • pp.67-76
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    • 2001
  • The "crescent-egg" a new spontaneous mutant was detected in a white egg strain k37. Studies were carried out the linkage analysis, to investigate phenotypic characteristics and biochemical analysis of haemolymph and ovarian protein. The mutant, ore was independent from 20 linkage groups P(2), Ze(3),L(4), oc(5), sn8), Ia(9), w-1(10), K(11), ch(13), U(14), bl(15), cts(16), bts(17), mln(18), nb(19), oh(20),Lan(21), or(22), tub(23) and Xan(27). The fertilization, hatchability and larval growth were not different from the those of normal eggs. The content and composition of yolk protein were similared to normal eggs. Scanning electron microscopy revealed the areal specific structure in dorsal region of egg-shell of cre mutant. Analysis of chorion protein by isoelectrofocusing(IEF), was resolved no difference in the composite of the chorion protein. We conclude that the egg mutant ere is expressed only in the egg-shape formation and region specific determination.

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Purification and Characterization of a Novel 21 kD Calcium Binding Protein from Dunaliella salina

  • Ko, Jae-Heung;Lee, Sun-Hi
    • Journal of Plant Biology
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    • v.39 no.3
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    • pp.173-177
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    • 1996
  • A novel calcium binding protein (CaBP) was purified to electrophoretic homogeneity from Dunaliella salina. In the course of purification experiment, this CaBP was identified as a monomer and its molecular weight was about 21 kDand isoelectric point (pI) value was about 4.1 using isoelectrofocusing. This CaBP was able to bind Ca2+ even in the pressence of an excess MgCl2 and KCI both in solution. In the SDS-PAGE, the Ca2+-bound form was slower than the Ca2+-free form in the nondenaturing PAGE. This means that the CaBP undergoes conformational change in the Ca2+-bound condition. Furthermore, UV absorption spectrum and fluorescence intensity of this CaBP was investigated. UV absorption peak was appeared at about 258 nm and decreased somewhat in Ca2+-bound condition. In the measurement of fluorescence, maximum intensity was appeared at 303 nm and decreased in Ca2+-bound state, similarly as UV absorption spectrum. These show distinct changes upon Ca2+-binding, which indicate of structural and/or dynamic changes largely reminiscent of other members of the EF-hand Ca2+-binding protein family.

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Two-Dimensional Electrophoretic Analysis of Rice Seed Proteins (쌀 종자 단백질의 2차원 전기영동적 분석)

  • Yoon, Hye-Hyun;Kim, Seung-Ho
    • Applied Biological Chemistry
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    • v.32 no.2
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    • pp.85-90
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    • 1989
  • High resolution two-dimensional (2-D) electrophoresis with isoelectrofocusing in the first dimension and electrophoresis in sodium dodecyl sulfate in acrylamide gradient gels in the second dimension has been used to produce maps of proteins, extracted from rice seeds with 2% sodium dodecyl sulfate/5% 2-mercaptoethanol. Six rice cultivars-three Japonica types and three Tongil(high-yielding) types-at six maturities were studied. Composite map was constructed and more than 300 polypeptide spots were counted in the pH range of $5.2{\sim}8.3$ and molecular range of $20,000{\sim}100,000$. Vast differences were observed between varieties and between maturities in the maps.

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Purification and Properties of Polyphenol Oxidase from Perillae Folium (들깨잎중의 폴리페놀 산화효소의 정제 및 특성)

  • 박수선;김안근;노진희;심미옥
    • YAKHAK HOEJI
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    • v.35 no.3
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    • pp.222-230
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    • 1991
  • Polyphenol oxidase (PPO) was purified from an extract of Perillae Folium by ammonium sulfate fractionation and sephadex G-150 gel filtration, which molecular weight estimated 65,000$\pm$1,000 in SDS-gel electrophoresis, and pI value was 4.8. The pH and temperature optima were 6.0 and $30^{\circ}C$ respectively. $K_{m}$ values of the PPO for various phenolics derived from Lineweaver-Burk plots were 4.0$\times$10$^{-4}$, caffeic acid; 4.2$\times$10$^{-3}$M, 4-methylcatechol. The inhibition by 4-nitrocatechoi, potassium cyanide, cysteine, 2-mercaptoethanol was competitive with $K_{i}$ values of 7.6$\times$10$^{-5}$M, 7.2$\times$10$^{-5}$M, 3.6$\times$10$^{-5}$M, 2.2$\times$10$^{-5}$M, respectively. Among the divalent cations, Cu$^{2+}$ ion was strong activator on PPO and Zn$^{2+}$, Ni$^{2+}$ ions were little effect on PPO activity. In comparing the amino acid composition of Perillae Folium PPO with that of wheat isozyme, grape, spinach showed similarity. But the content of glycine phenylalanine was abundant relatively.

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Direct Detection of (1-3)-$\beta$-Glucanase Isozymes in Isoelectrofocusing Gels Using a Dye -Labeled Substrate (염료착색 기질을 이용한 IEF gel에서(1-3)-$\beta$-glucanase 동위효소의 검출)

  • Yun, Song-Joong;Lee, Myong-Chul;Kwon, In-Sook;Kim, Tae-San;Go, Seung-Joo
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.39 no.2
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    • pp.121-127
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    • 1994
  • A procedure for the direct detection of (1-3)-$\beta$-glucanase isozymes in electrophoresis gels was developed. The procedure employed the commercial preparation of AZCL-pachyman as a chromogenic substrate for (1-3)-$\beta$-glucanases. The procedure detected the three basic isozymes which have been known to be expressed in germinating barley kernels. A major acidic and a minor isozymes were also detected in germinating kernels. The procedure was proved to be fast, simple and sensitive enough to be used for the analysis of the expression of (1-3)-$\beta$-glucanase isozymes in plant tissues. The detection limit of the procedure for the commercial preparation of Penicillium (1-3)-$\beta$-glucanase was estimated to be as low as 50$\mu$U. The procedure could be used for the investigation of (1-3)-$\beta$-glucanases in laboratories facilitated with ordinary equipments and research personnel.

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The Biocontrol Activity of Chromobacterium sp. Strain C-61 against Rhizoctonia solani Depends on the Productive Ability of Chitinase

  • Park, Seur-Kee;Lee, Myung-Chul;Harman, Gary E.
    • The Plant Pathology Journal
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    • v.21 no.3
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    • pp.275-282
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    • 2005
  • A chitinolytic bacterium, Chromobacterium sp. strain C-61, was found strongly antagonistic to Rhizoctonia solani, a causal agent of damping-off of eggplant. In this study, the biocontrol activity and enzymatic characteristics of strain C-61 were compared with its four Tn5 insertion mutants (C61-A, -B, -C, and -D) that had lower chitinolytic ability. The chitinase activity of a 2-day old culture was about $76\%,\;49\%\;and\;6\%$ level in C61-A, C61-B and in C61-C, respectively, compared with that of strain C-61. The $\beta-N-acetylhexosaminidase$(Nahase) activity was little detected in strain C-61 but increased largely in C-61A, C61-B and C61-C. Activities of chitinase and Nahase appeared to be negatively correlated in these strains. Another mutant, C-61D, produced no detectable extracellular chitinase and Nahase. The in vitro and in vivo biocontrol activities of strain C-61 and its mutants were closely related to their ability to produce chitinase but not Nahase. No significant differences in population densities between strain C-61 and its mutants were observed in soil around eggplant roots. The results of SDS-PAGE and isoelectrofocusing showed that a major chitinase of strain C-61 is 54-kDa with pI of approximately 8.5. This study provides evidence that the biocontrol activity of Chromobacterium sp. strain C-61 against Rhizoctonia solani depends on the ability to produce chitinase with molecular weight of 54-kDa and pI of 8.5.

A Systematic Study on the Selected Amentiferous Plant Taxa -On the Genus Quercus- (유이화서 식물군의 계통분류학적 연구 -참나무과를 중심으로-)

  • Lee, Yoo Sung
    • Journal of Plant Biology
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    • v.27 no.2
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    • pp.61-72
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    • 1984
  • The concept of natural grouping of plant designated as the "Amentiferous" is no longer given serious credence, and many of the families included in this grouping have been dispersed in diverse order. Because a review of taxonomic treatments of amentiferous taxa reveals diverse classifications, it has become necessary to investigate new characteristics and attempt to determine the significance of these characteristics in terms of amentiferous taxonomy. Protein analyses by isoelectrofocusing(IEF) and rocket immunoelectrophoresis(RIE) have proved to be useful in the delicitation of Quercus taxa using pollen extracts from selected taxa. When Quercus pollen extracts were separated by electrophoresis based on their isoelectric points in a stable pH gradient and substrates for estrase activity were stained, ten bands were revealed between pH 5-14. Within Lepidobalanus grouping, a great diversity in the pollen protein zymograms was observed with some segregation corresponding to the designated taxonomic sections. Two taxa of Cyclobalanopsis produced a zymogram that is somewhat similar to taxa included within the section Prinus of Lepidobalanus, and less similar to taxa within the section Cerris of the same subgenus. Three tested taxa of the Cerris are in the similar zymogram each other, being segregable from the taxa of Prinus. Quantitative and qualitative analyses for serological relationships within and among th Quercus were also employed. To calculate the degree of protein similarity, total rocket heights obtained from RIE provided an index of serological correspondence(SC). It is reconfirmed that the Quercus is distantly separated from the Fagus according to SC. Comparative data from rocket number and SC in the tested taxa of Quercus also indicate that Lepidobalanus is separable from Cyclobalanopsis. Within the Lepidobalanus Q. acutissima and Q. acutissima x variabilis are almost homogeneors and distinguishable from the other tested taxa of same subgenus. Although the number of taxa tested has been limited, the overall serological evidence best reflects the classification proposed by Redher(1940) and Melchior (1964), having the genus Quercus subdivided into three subgunera: Erythrobalanus, Lepidobalanus, and Cyclobalanopsis.alanopsis.

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Effect of Gibberellic acid on Isozyme Pattern of Rice Plant (Gibberellic acid가 수도의 Isozyme pattern에 미치는 영향)

  • Park, W.M.;Lee, Y.S.;Son, E.R.
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.29 no.1
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    • pp.39-45
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    • 1984
  • The present researches were carried out to investigate the effects of gibberellic acid on the appearance of isozyme patterns of esterase, phosphatase, amylase and peroxidase, also to investigate if there were any differences of the isozyme patterns among root, shoot and seeds of rice plants. It was noticed that the isozyme patterns of the above tested enzymes were differ among the organs, root, shoot and seed. The GA treated plants showed difference of esterase patterns in root from Akibare and the difference in shoot and root from Yushin, phosphatase patterns in root from Akibare. However, the GA did not affect isozyme patterns of amylase or peroxidase. The seed should be the suitable organ to study isozyme patterns for genetics or variety characterization of rice plant.

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