• Title/Summary/Keyword: Protein Isozyme patterns

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Superoxide Dismutase and Peroxidase Activity of Transformed Callus in Tomato (형질전환된 토마토 캘러스의 Superoxide Dismutase와 Peroxidase 활성)

  • 유정민;정형진;김경민;곽상수
    • Korean Journal of Plant Tissue Culture
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    • v.25 no.3
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    • pp.177-181
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    • 1998
  • This study was carried out to investigate activity difference in the superoxide dismutase (SOD) and peroxidase (POD) of tomato callus transformed with Agrobacterium containing the GUS gene. Than those of other two tomato cultivars, the hypocotyl explant of JA101 was shown to have higher POD and SOD specific activity of 23 unit/mg protein and 2,156 unit/mg protein, respectively. Relatively high frequency of callus formation was obtained from the hypocotyl explant on MS medium containing 1 mg/L 2,4-D for 30 days and its POD(47 unit/mg protein) and SOD (95,786 unit/mg protein) specific activities were higher than other 2,4-D concentration. The hypocotyl explant and callus cocultivated with Agrobacterium for 72 hours were transferred to MS medium supplemented with 1 mg/L 2,4-D, 30 mg/L kanamycin, 30 g/L sucrose and 4 g/L Gelrite. The hypocotyl explants transferred to the medium formed callus with 45.5% effeciency after 8 weeks. The transformation efficiency confirmed by GUS assay was 21.6%. POD specific activity of the transformed callus (54 unit/mg protein) were somewhat lower than the non-transformed callus (64 unit/mgg protein) and SOD specific activity of the transformed callus (30,300 unit/mg protein) were also lower than the non-transformed callus (37,077 unit/mg protein). However there was no significant difference in POD and SOD isozyme patterns between the transformed and the non-transformed calluses. From these results, it revealed that there was no difference of antioxidant enzyme activities between the transformed callus and the non-transformed callus in tomato.

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Application of Electrophoretic Methods for differentiation of Trichoderma species (전기영동법을 이용한 Trichoderma spp 분류가능성에 관한 연구)

  • Park W.M.;Park Y.H.;Lee E.Y.
    • Korean journal of applied entomology
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    • v.23 no.2 s.59
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    • pp.102-108
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    • 1984
  • These researches were carried out to investigate the morphology of different species of Trichoderma and the possibilities of differentiation of the species of Trichoderma by electrophoretic methods. Variations between the isolates of a species of Trichoderma indicate the genetical differences, also isozyme and protein patterns will be useful to investigate genetical variations betweens the isolates. It might be possible that distinct bands of isozymes of esterase, phosphotase, catalase, catalase differentiate species of Trichoderma.

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Extraction of Intercellular Material from Rice Leaf Tissue (수도의 세포간극 물질 추출)

  • Park, W.M.;Son, E.R.;Ko, Y.H.;Yoo, Y.J.;Lee, Y.S.
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.28 no.3
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    • pp.323-327
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    • 1983
  • The intercellular material was extracted from rice leaf tissue. The quantitative tests of protein and soluble carbohydrate, and activity of peroxidase, showed differences between tissue extract and intercellular material. Also electrophoretic patterns of peroxidase and esterase isozymes were not similar between them. It indicated that the intercellular material which was extracted, was not the mixture of cellular materials.

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Isozyme Variability in Three Species of Freshwater Planorbid Snails in Korea : Gyraulus convexiusculus , Hippeutis cantori and Segmentina hemisphaerula (한국산 또아리물달팽이과 ( family Planorbidae ) 3종에서의 동위 효소 변이)

  • 정평림;정영헌;김기선
    • The Korean Journal of Malacology
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    • v.11 no.1
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    • pp.51-61
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    • 1995
  • A horizontal starch gal electrophoresis for enzyme proteins extracted from 3 species of Korean planorbid snails; Gyraulus convexiusculus, Hippeutis cantori and Segmentina hemisphaerula was carried out in order to elucidate their genetic relationships.The results from 12 enzymes employed in three different kinds of buffer systems are summarized as follows:1) Two loci from each enzyme of aldehyde oxidase, esterase, glucose phosphate isomerase. isocitrate dehydrogenase, leucine aminopeptidase, malate dehyogenase, peptidase and xanthine oxidase were detected, and only one locus was observed from each of the following four enzymes: 6-phosphogluconate dehydrogenase, glyceraldehyde-phosphate dehydrogenase, glutamate oxaloacetate transaminase and glycerol-3-phosphate dehydrogenase.2) Most of loci in 3 species of planorbid snails employed showed homozygous and monomorphic banding patterns and some of them were specifis as genetic markers among different species. However, a few of loci (EST-1. EST-2 and GPI-2)showed polymorphic banding patterns. 3)Hippeutis cantori and Segmentina hemisphaerula were more closely clustered in a dendrogram with the genetic iddentity value of 0.431, and these two species were lineated with Gyraulus convexiusculus as another cluster at the value of 0.294.In summarizing the above results, three species of Korean planorbid snails employed in this study mostly showed monomorphic enzyme protein banding patterns and genetic differences specific among 3 species.

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Effects of Ethephon and Gibberellin on Sex Expression, and Subsequent Changes in Protein Contents, Peroxidase Activities, and Isoperoxidase Isoperoxidase Patterns of Cucumis sativus L. (Ethephon과 Gibberellin 처리(處理)가 오이의 성발현(性發現)과 이에 따른 단백질함량(蛋白質含量), Peroxidase 활성(活性) 및 Isoperoxidase Pattern에 미치는 영향(影響))

  • Ku, Woo Seo;Kim, Young Rae
    • Korean Journal of Agricultural Science
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    • v.12 no.1
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    • pp.1-16
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    • 1985
  • This experiment was conducted to study sex expression and subsequent changes in protein and peroxidase after three cultivars of cucumber plants were treated with ethephon and gibberellin. The three cultivars of cucumber used in this study included 'Sayeup' (monoecious type), 'Sinrokdadaki' (gynoecious type), and 'Seonghowon' (intermediate type). The ethephon at 250 ppm and gibberellin at 100 ppm were treated at the 2-leaf and 4-leaf stages, and subsequent sex expression and changes in protein contents, peroxidase activities and isoperoxidase banding patterns by disc polyacrylamide gel electrophoresis were studied. The results can be summarized as follows: 1. Ethephon treatment slightly increased number of pistillate flowers and significantly decreased number of staminate flowers in the three cultivars, while gibberellin treatment significantly increased number of staminate flowers in both gynoecious 'Sinrokdadaki' and intermediate 'Seonghowon' and did not increase number of staminate flowers in monecious 'Sayeup' 2. There were some differences among three cultivars in protein contents, protein banding and isoperoxidase banding patterns of seeds and germinating seeds. However, it was not obvious to differentiate monoecious from gynoecious cultlvars by these characters. 3. Protein contents in the leaves. and stem apex after ethephon and gibberellin treatment increased gradually at the 2-leaf stage, but decreased at the 4-leaf stage. Protein contents in stem apex at the 4-leaf stage without treatment were much higher in 'Sinrokdadaki' and 'Seonghowon' than in monoecious 'Sayeup'. Protein contents in the stern apex at the 4-leaf stage were increased in the ethephon-treated monoecious 'Sayeup' and decreased in 'Sinrokdadaki' and 'Seonghowon' compared with untreated plants. 4. Peroxidase activities in the leaves and stem apex gradually decreased at the 2- leaf stage, but increased at the 4-leaf stage. Peroxidase activities in stern apex at the 4-leaf stage were significantly increased by ethephon treatment. 5. The number of protein bands in the three cuitivars after treatment gradually decreased in leaves and stem apex at the 2-leaf stage, but increased in the 4-leaf stage. The protein banding pat terns of stern apex of the ethephon-treated monoecious 'Sayeup' at the 4-leaf stage were gradually shifted to the banding.

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Phenoloxidases and Photomorphogenesis in Coprinus congregatus (Coprinus congregatus의 분화와 Phenoloxidase와의 관계)

  • 최형태
    • Proceedings of the Botanical Society of Korea Conference
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    • 1987.07a
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    • pp.157-167
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    • 1987
  • The have been many reports that phenoloxidase are correlated with development in many fungi. C. congregatus, one of nushroom-forming basidiomycetes, which requires light for its development also has phenoloxidases. In C. congragatus, there are two sets of membrane-associated phenoloxidase (PHO I and PHO II) which are differentiated by their isozyme patterns, and each enzyme set consists of two different subtrate specific enzyme protein; o-tolidine reacting enzyme, and DOPA reacting enzyme. PHO I which is localized by a protoplast-concanavalin A technique by using a new solidifying agent, Pluronic Polyol F 127, instead of agar appears in the vegetative hyphae, and PHO II appears at the early primordial stage on agar and at the sclerotial stage of liquid shake cultures. Inhibition of PHO I with the enzyme inhibitors inhibits mushroom formation as well as melanization of the vegetative hyphae at concentrations which do not inhibit the vegetative growth. PHO I deficient mutants do not form mushrooms or melanins, and the mutants show abnormal nuclear migration patterns. PHO II has roles; possibly cementing the adjacent hyphae during the actual three dimensonal structure formation, and melanizing mushrooms and sclerotia. The possible roles of PHO I in the light reception complex and in melanin formation, the function of malanin, and possible roles of postulated post translational modifying enzymes which regulate the phenoloxidases, nuclear migration pattern, and self-nonself recognition mechanism are discussed.

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Studies on the Domestication of of Field Bean ( Glycine soja Sieb and Zucc ) for Forage Crop I. Esterase isozyme variation and nutritive value of field bean (돌콩 ( Glycine soja Sieb and Zucc ) 의 사료작물화에 관한 연구 I. 돌콩의 Esterase isozyme 변이와 사료적 가치 평가)

  • Lee, Sung K.;Lee, Eun;Choi, Il
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.13 no.2
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    • pp.86-92
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    • 1993
  • The variation of the esterase isozyme, germination rate, chemical composition and digestibility of field bean(G1ycine soja S. and Z.) were estimated. The results are as follows; 1. The banding patterns of the esterase isozyme in field bean were varied with the tissue and habitat. 2. The enzyme activity of the Est-I, Est-2, Est-3 and Est-4 in field bean showed a high value compared with the other enzyme. 3. The range of germination temperature in field bean was 10-40C and the optimum germination temperature was 25- 38^{\circ}C.$. 4. The crude protein(CP) contents was 19.9% in the whole plant, 27.8% in the leaf and 45.9% in the seed, the cellulose contents was 29.5% in the whole plant, 23.0% in the leaf and 13.8% in the seed, the neutral detergent fiber(NDF) was 62.6% in the whole plant, 47.9% in the leaf and 47.9% in the seed and the acid detergent fiber(ADF) was 44.5% in the whole plant, 28.4% in the leaf and 28.4% in the seed, respectively. 5. The digestibility of the field bean was 44.1% in the whole plant, 49.6% in the leaf and 75.1% in the seed, NDF was 26.2% in the whole plant 46.2% in the leaf, ADF was 29.0% in the whole plant, 47.7% in the leaf and 58.0% in the seed and Cellulose was 48.7% in the whole plant, 58.0% in the leaf and 70.2% in the seed, respectively. 6. Total digestible nutrients(TDN) of the field bean was 47.4% in the whole plant, 51.5% in the leaf and 70.2% in the seed, respectively. The digestible energy(DE) value was 2.1 kcal/g in the whole plant, 2.27 kcal/g in the leaf and 3.10 kcal/g in the seed and the metabolizable energy(ME) value was 1.72 kcal/g in the whole plant, 1.86 kcal/g in the leaf and 3.23 kcal/g in the seed, respectively.

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Improvement of Peroxidase Productivity by Optimization of Medium Composition and Cell Inoculum Size in Suspension Cultures of Sweet Potato (Ipomoea batatas) (고구마(Ipomoea batatase)현탁배양에서 배지조성 및 세포접종량의 적정화에 의한 Pemxidase생산성 향상)

  • 곽상수;김수경;정경희;유순희;박일현;유장렬
    • Korean Journal of Plant Tissue Culture
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    • v.21 no.2
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    • pp.91-97
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    • 1994
  • To improve the productivity of peroxidase (POD) of cell line SP-47 derived from cell suspension cultures of sweet potato (Ipomoea batatas (L) Lam.cv White Star), we optimized culture conditions including the composition and concentration of plant growth regulators and carbon source, and the cell inoculum size. When one g (fr wt) of cells was inoculated into 50 mL TL medium supplemented with l mg/L 2,4-D and 30g/L sucrose in 300 mL Erlenmeyer flask at 25$^{\circ}C$ in the dark (100rpm), the POD activity per g cell dry wt was maximized to be about 6,800 units after 25 days of subculture, which was about 30 times higher than that of intact roots of horseradish plants grown in the greenhouse, but the cell growth was maximum after 15 days of subculture. The protein content per g cell dry wt maintained almost plateau and after 25 days of subculture decreased as culture Proceeded further whereas the POD specific activity (unit/mg protein) was about two times higher after subculture and continuously increased from 12 days to the end of cultures (40 days). The POD isozyme patterns showed almost the same regardless of cell growth stage, but some acidic isozymes were slightly increased after 25 days of subculture. These results indicate that POD activity in suspension cultures of sweet potato is closely associated with cell growth and stresses derived from cell culture renditions and medium depletion. Due to its high POD activity the SPL47cell line seems to be suitable for the mass production of POD.

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Purification and Characterization of Lactate Dehydrogenase Isozymes in Channa argus (가물치(Channa argus) 젖산탈수소효소 동위효소들의 정제 및 특성)

  • Park, Eun-Mi;Yum, Jung-Joo
    • Journal of Life Science
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    • v.20 no.2
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    • pp.260-268
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    • 2010
  • The lactate dehydrogenase (EC 1.1.1.27, LDH) isozymes in tissues from Channa argus were purified and characterized by biochemical, immunochemical and kinetic methods. The activity of LDH in skeletal muscle was the highest at 380.4 units and those in heart, eye and brain tissues were 13.4, 3,5 and 5.4 units, respectively. Citrate synthase (EC 4.1.3.7, CS) activity in heart tissue was the highest at 20.7 units. LDH/CS in skeletal muscle, heart, eye and brain tissues were 172.9, 0.6, 0.32 and 0.47. Protein concentration in skeletal muscle tissue was 14.7 mg/g and specific activities of LDH in skeletal muscle, heart, eye and brain tissues were 25.88, 0.79, 0.31 and 1.38 units/mg, respectively. Therefore, skeletal muscle tissue was anaerobic and heart tissue was aerobic. The LDH isozymes in tissues were identified by polyacrylamide gel electrophoresis, immunoprecipitation and Western blot with antiserum against $A_4$, $B_4$, and eye-specific $C_4$. LDH $A_4$, $A_3B$, $A_2B_2$. $AB_3$ and $B_4$ isozymes were detected in every tissue, $C_4$, $AC_3$, $A_2C_2$ and $A_3C$ were detected in eye tissue, and $A_3C$ was found in brain tissue. LDH $A_4$, $A_3B$, $A_2B_2$, $AB_3$, $B_4$, eye-specific $C_4$ isozymes were purified by affinity chromatography and Preparative PAGE Cells. The LDH $A_4$ isozyme was purified in the fraction from elution with $NAD^+$ containing buffer of affinity chromatography. Eye-specific $C_4$ isozyme was eluted right after $A_4$, after which $B_4$ isozyme was eluted with plain buffer. As a result, one part of molecular structures in $A_4$, $B_4$ and eye-specific $C_4$ were similar, but were different from each other in $B_4$ and $C_4$. Therefore the subunit A may be conservative in evolution, and the evolution of subunit B seems to be faster than that of subunit A. The activity of LDH $A_4$, $A_2B_2$, $B_4$, and eye-specific $C_4$ isozymes remained at 39.98, 21.28, 19.67 and 16.87% as a result of the inhibition by 10 mM of pyruvate, so the degree of inhibition was very high. The $Km^{PYR}$ values were 0.17, 0.27 and 0.133 mM in $A_4$, $B_4$ and eye-specific $C_4$ isozymes, respectively. The optimum pH of LDH $A_4$, $B_4$, eye-specific $C_4$, $A_2B_2$, $A_3B$, and $AB_3$ were pH 6.5, pH 8.5, pH 5.5, pH 6.0-6.5, pH 5.0 and pH 7.5. The $A_4$ and heterotetramer isozymes stabilized a broad range of pH. Especially, LDH activities in skeletal muscle tissue were high, resulting in a high degree of muscle activity.LDH metabolism in eye tissue seems to be converted faster from pyruvate to lactate by eye-specific $C_4$ isozyme as eye-specific $C_4$ have the highest affinity for pyruvate, and right after the conversion, oxidation of lactate was induced by $A_4$ isozyme. It was found that expression of Ldh-C, affinity to substrate and reaction time of $C_4$ isozyme were different according to the ecological environmental and feeding capturing patterns.

Studies on Constituents of Higher Fungi of Korea (LXXI) -Application of Enzymes to Taxonomy of Ganoderma Species-

  • Kim, Byong-Kak;Kim, Jin-Sook;Choi, Kyun-Gae;Kim, Ha-Won;Choi, Eung-Chil
    • Korean Journal of Pharmacognosy
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    • v.24 no.2
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    • pp.116-123
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    • 1993
  • The genus Ganoderma is typical wood-rotting fungi and its fruiting body has been used as an important herb in oriental medicine. Recent research discovered antitumor components from Ganoderma lncidum. Various Ganoderma species are being cultivated in Korea. However, taxonomic system of the genus Ganoderma has been based mainly on the macromorphology of fruiting bodies and the ultrastructural characteristics of basidiospores. Since there are similar characteristics in Ganoderma mycelia grown on the same artificial media, it is suggested that the compatibility of the fungi by di-mon mating be used as an aid to determine the identity of species in addition to the conventional characterization. In this study, we examined physiological and genetical properties such as growth temperature, pH, compatibility and enzyme or protein patterns of laccase, esterase and cellular proteins of G. lucidum RZ, G. tsugae and Ganoderma species cultivated in Korea by electrophoresis for characterization of the isolates. We found that compatibility test and isozyme patterns of laccase and esterase of the mycelia could be used for the differentiation of the isolates. These results showed that Ganoderma species cultivated in Korea is genetically similar to G. lucidum but physiologically closer to G. tsugae than to G. lucidum.

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