• 제목/요약/키워드: isozyme

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Electrochemical properties of the mugwort-embedded biosensor for the determination of hydrogen peroxide (쑥을 이용한 과산화수소 정량 바이오센서의 전기화학적 성질)

  • Lee, Beom-Gyu;Park, Sung-Woo;Yoon, Kil-Joong
    • Analytical Science and Technology
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    • v.19 no.1
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    • pp.58-64
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    • 2006
  • A mugwort-tissue-based modified carbon paste electrode was constructed for the amperometric detection of hydrogen peroxide and its electrochemical properties are described. Especially the amperometric signal was very stable and bigger than any other enzyme electrode studied in this lab. The effect of tissue composition on the response was linear within the wide range of experiment and the linearity of Lineweaver-Burk plot showed that the sensing process of the biosensor is by enzymatic catalysis. And pH dependent current profile connoted that two isozymes are active in this system.

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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Purification and Properties of Branched Chain Amino Acid Arminotransferase from Fasciola hepatica (간질(Fasciola hepatica)의 Branched Chain 아미노산 Aminotransferase의 정제 및 성상)

  • 이중호;이동욱이의성송철용
    • Parasites, Hosts and Diseases
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    • v.21 no.1
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    • pp.49-57
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    • 1983
  • The distribution and Properties of branched chain amino acid aminotransferase (EC 2.6. 1.42) was investigated in adult Fasciola hepatica. Fascicla hepatica was fractionated by differential centrifugation into nuclear, mitochondrial and cytosolic fractions. The activity of branched chain amino acid aminotransferase was measured by the method of Ichihara and Koyama (1966) . Isozyme patterns of this enzyule was also examined by DEAE-cellulose column chromatography. The results obtained were as follows; 1. The activity in homogenate was found to be 12.69 units/g wet tissue. The activity of this enzyme was relatively high compared with those in rat tissues. 2. The distribution of branched chain amino acid aminotransferase in the subcellular organelles showed that 87.8% of the activity was in cytosolic, 10.9% in mitochondrial and 1.3% was in nuclear fraction. 3. Cytosolic fraction of Fasciola hepatica contained Enzyme I, but not Enzyme II and III, of branched chain amino acid aminotransferase. Ensyme I was eluted by 50mM phosphate buffier from DEAE-cellulose column and catalyzed the transamination of all three branched chain amino acids. 4. The Enzyme I was purified about 22-folds increase in specific activity after chromatography on DEAE-cellulose. 5. The best substrate among three amino acids (leucine, isoleucine and valise) was L-isoleucine. 6. The optimal temperature of Enzyme I was $45^{\circ}C$ and the optimal pH was 8.2. 7. The Km value for leucine of Enzyme I was 4.17 mM. 8. The Km values for a-ketoglutarate and pyridoxal phosphate of Enzyme I were 0.41mM and $4.76{\times}10^{-3}{\;}mM$, respectively.

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Genetic Diversify and Population Structure of Two Korean Pond Frog Species, Rana nigromaculata and R. plancyi (Anura, Ranidae), with a Survey of Temporal Genetic Variation in R. nigromaculata

  • Suh-Yung Yang;Jong-Bum Kim;Mi-Sook Min;Jae-Hwa Suh
    • Animal cells and systems
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    • v.3 no.3
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    • pp.275-283
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    • 1999
  • Korean R. plancyi occupies a restricted area in western South Korea and shows a relatively low level of genic variability (%P=15.2, Ho=0.052, He=0.048). In contrast, R. nigromaculata is broadly distributed in South Korea. The observed low level of variability of R. nigromaculata (%P=14.3, Ho=0.042, He=0.043) is probably due to its recent colonization. Populations of R. nigromaculata exhibited considerable genetic differentiation (F$_{sT}$=0.149) and low level of gene flow (Nm=1.427) among populations, compared to those of R. Plancyi (F$_{sTF$_{sT}$}$=0.096, Nm=2.354), which occupies a restricted area. The observed levels of gene flow among populations of R. nigromaculata (Nm=1.427) over a broad geographic range is relatively higher than other amphibian species. The high level of gene flow is probably the result of the high dispersal abilities of R. nigromaculata. A survey of temporal genic variation of R. nigromaculata showed that there was no significant change on the overall average genetic diversity from 1978 (average He=0.044) to 1997 (average He=0.040). Wright's F-statistics also indicated no significant genetic differentiation from 1978 (F$_{sT}$=0.118) to 1997 (F$_{sT}$=0.108). This suggests that the environmental change appears to have had little influence on the genetic composition of R. nigromaculata in the study areas during the past 20 years. The low level of temporal variation might be due to the result of high dispersal abilities and wide migration range of this species.

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Comparison of Glucuronidating Activity of Two Human cDNAs, UDPGTh1 and UDPGTh2

  • Kim, Soon-Sun;Owens, Ida-S.;Sheen, Yhun-Yhong
    • Archives of Pharmacal Research
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    • v.20 no.5
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    • pp.454-458
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    • 1997
  • Two human liver UDP-glucuronosyltransferase cDNA clones, HLUG25 and UDPGTh2 were previously shown to encode isozymes active in the glucuronidation of hyodeoxycholic acid (HDCA) and certain estrogen derivatives (e.g., estriol and 3,4-catechol estrogens), respectively. in this study we have found that the UDPGTh2-encoded isoform (UDPGTh2) and HLUG25-encoded isoform (UDPGThl) have parallel aglycone specificities. When expressed in COS 1 cells, each isoform metabolized three types of dihydroxy- or trihydroxy-substituted ring structures, including the 3,4-catechol estrogen (4-hydroxyestrone), estriol, 17-epiestriol, and HDCA, but the UDPGTh2 isozyme was 100-fold more efficient than UDPGTh1. UDPGTh1 and UDPGTh2 were 86% identical overall (76 differences out of 528 amino acids), including 55 differences in the first 300 amino acids of the amino terminus, a domain which conferred the substrate specificity. The data indicated that a high level of conservation in the amino terminus was not required for the preservation of substrate selectivity. Analysis of glucuronidation activity encoded by UDPGTh1/UDPGTh2 chimeric cDNA constructed at their common restriction sites, Sac I (codon 297), Nco I (codon 385), and Hha I (codon 469), showed that nine amino acids between residues 385 and 469 were important for catalytic efficiency, suggesting that this region represented a domain which was critical for the catalysis but distinct from that responsible for aglycone-selection. These data indicate that UDPGTh2 is a primary isoform responsible for the detoxification of the bile salt intermediate as well as the active estrogen intermediates.

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Isolation and Cloning of Porcine SLC27A2 Gene and Detection of Its Polymorphism Associated with Growth and Carcass Traits

  • Wang, Tao;Liu, Chang;Xiong, Yuan-Zhu;Deng, Chang-Yan;Zuo, Bo;Xie, Hong-Tao;Xu, De-Quan
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.8
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    • pp.1169-1173
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    • 2007
  • The protein encoded by SLC27A2 gene is an isozyme of long-chain fatty-acid-coenzyme A ligase family, and it converts free long-chain fatty acids into fatty acyl-CoA esters, and thereby plays a key role in lipid biosynthesis and fatty acid degradation. In the present study, SLC27A2 located on human chromosome 15 was selected as candidate gene and we isolated and cloned partial fragments of mRNA sequence and genomic fragments of porcine SLC27A2 gene. The coding region of the gene as determined by alignments shared 90% and 82% identity with human and mouse cDNAs, respectively. Detection in LargeWhite and Meishan breeds showed that a single nucleotide polymorphism (SNP) ($A{\rightarrow}G$) existed in exon 7, which caused corresponding amino acid changed for encoding. In LargeWhite pigs it encoded for Val while in Meishan pigs it encoded for Ile, so we developed the PCR-RFLP genotype method for detection of this polymorphism. Association study in 135 $F_2$ reference family indicated that significant correlation existed between the polymorphism and growth and carcass traits.

Systematics and Evolutionary Study of the Family Corvidae(Passeriformes) in Korea (한국산 까마귀과(참새목) 조류의 계통 진화)

  • 심재한;박병상;양서영
    • Korean Journal of Environment and Ecology
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    • v.11 no.3
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    • pp.277-289
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    • 1997
  • Electrophresis methods were used to examined the degree of genetic relatedness and divergent times on the 4 genera 4 species of the family Corvidae and 1 species of the Family Cinclidae. Genetic relatedness between two families and between 4 genera of Corvidae showed similar to those reported at comparable taxonomic levels in other bird. But it appears to be considerably less than that of non avian taxa. Genetic relatedness between Garrulus glandarius brandtii and Corvus corone orientalis was the most close related(S=0.81), whereas Pica pica sericea(S=0.72) and Cyanopica cyanus koreensis(S=0.68) were showed less close relatedness respectively. The presumed divergent time between two families was 3.9 million years before present. Whereas among the 4 species of corvidae, the presumes divergent times of Cyanopica cyanus koreensis and Pica pica sericea were about 1.9 and 1.6 million years before present respectively, and between Garrulus glandarius brandtii and Corvus corone orientalis was showed recently about 1.0 million years before present

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Genetic Variation of Several Isoenzymes in Pinus densiflora for. multicaulis (반송(Pinus densiflora for. multicaulis)의 몇 가지 동위효소(同位酵素)의 유전변이(遺傳變異))

  • Hwang, Jae Woo;Lee, Seok Woo
    • Journal of Korean Society of Forest Science
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    • v.85 no.3
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    • pp.409-415
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    • 1996
  • Isozyme variations in 4 enzyme systems in Pinus densiflora for. multicaulis in 31 individual trees collected throughout the country were studied by starch gel electrophoresis using haploid megagametophyte tissue to compare with those of Pinus densiflora. A minimum of 7 loci were found to code for isozymes of the enzyme systems. No variation was found at locus GOT-A; at the remaining 6 variable loci(GDH-A, GOT-B, GOT-C, IDH-A, LAP-,A, LAP-B) and 2 to 4 alleles were identified. We could not find any marker alleles to distinguish Pinus densiflora for. multicaulis from Pinus densiflora at the isozymes studied here. Allele frequency distributions at each loci were almost all the same as those of P. densiflora. The percentage of polymorphic loci(99% level), the number of alleles per locus, the observed and expected heterozygosities were 85.7, 2.3, 0.165 and 0.186%, respectively. The level of genetic diversity in Pinus densiflora for. multicaulis seemed to be less than that of P. densiflora.

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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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Determination of Seed Purity in Radish (Raphanus sativus L.) Using Allozyme (알로자임에 의한 무 씨의 순수성 검증)

  • Huh, Man-Kyu
    • Journal of Life Science
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    • v.18 no.7
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    • pp.907-911
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    • 2008
  • Radish (Raphanus sativus L.) is one of very important crop plants in the world. It is very important to determine hybrid seed quality in the production of hybrid Brassica vegetable seeds to avoid unacceptable contamination with self-inbred (sib) seeds. The allozyme for evaluating seed purity in a commercial $F_1-hybrid$ radish cultivar is demonstrated. Three hundred sixty seeds from the male and female harvest were subsequently screened for seed purity using 27 isozyme loci. Especially, F1 hybrids of radish, Per-1 ($aa{\times}bb$), Lap-1 ($aa{\times}bb$), Est-1 ($aa{\times}bb$) were presented clear hybrid bands. Est-1 locus revealed that 15 (8.3%) seeds from the female harvest and 26 (14.4%) seeds from the male harvest were sibs. It maintains higher than average level of genetic diversity compared with their correspondent parents. Shannon's index of phenotypic diversity (I) of hybrids was the highest of all accessions (R. sativus L. cv. Daepeng, R. sativus L. cv. Backza, and their hybrids). The allozyme may lead to a better insight into the hybrid seed purity.