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Characterization of Lactate Dehydrogenase in Acanthogobius hasta (풀망둑(Acanthogobius hasta) 젖산탈수소효소의 특성)

  • Yum, Jung-Joo
    • Journal of Life Science
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    • v.18 no.2
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    • pp.264-272
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    • 2008
  • The lactate dehydrogenase (EC 1.1.1.27, LDH) isozymes in tissues from Acanthogobius hasta were characterized by biochemical, immunochemical and kinetic methods. The activities of LDH in skeletal muscle and eye tissues were 65.30 and 53.25 units, but LDH activities in heart and liver tissues were very low. LDH/CS (EC 4.1.3.7, citrate synthase) in skeletal muscle was the highest as 22.29. Specific activities of LDH in brain, eye and skeletal muscle were 56.45, 38.04 and 11.0 units/mg, respectively. The LDH isozymes in tissues were separated by polyacrylamide gel electrophoresis after immunoprecipitation with antiserum against $A_4,\;B_4$ eye-specific $C_4$ and liver-specific $C_4$. LDH $AC_4$ isozymes were detected predominantly in skeletal muscle, brain and eye tissues, and $B_4$ isozyme was detected in heart. Anodal eye-specific $C_4$ and cathodal liver-specific $C_4$ were coexpressed in A. hasta. The eye-specific $C_4$ isozyme showed higher activity in eye tissue, but liver-specific $C_4$ isozyme showed lower activity in liver. As a result, one part of molecular structures in $A_4\;and\;C_4,\;A_4\;and\;B_4$, and eye-specific $C_4$ and liver-specific $C_4$ were similar, but in $B_4\;and\;C_4$ were different with each other. 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 LDH $A_4$ isozyme of skeletal muscle was purified in the fraction from elution with NAD+ containing buffer of affinity chromatography and eye-specific $C_4$ isozyme was eluted right after $A_4$, so the structure of eye-specific $C_4$ isozyme is similar to $A_4$. And LDH activity remained 35.22-43.47% as a result of the inhibition by pyruvate, the Michaelis-Menten constant values for pyruvate was 0.080-0.098 mM, and Vmax were 153.85 units, 35.09 units in skeletal muscle and eye, respectively. Also the $B_4$ isozyme was the thermo-stablest and $C_4$ was stabler than $A_4$ isozyme. The optimum pH of LDH was 6.5. The results mentioned above indicate that isozymes in tissues showed the properties between LDH $A_4\;and\;B_4$ isozyme as A. hasta was adapted to hypoxic conditions. Also LDH seems to function more effectively under anaerobic condition because LDH in skeletal muscle and eye tissues have high affinity for pyruvate.

Peroxidase Isozyme Pattern and Polyamine Contnts in Germinating Peas (Pisum staivum) (완두 발아시 Polyamine 함량 변화 및 Peroxidase Isozyme 양상)

  • 표병식
    • Journal of Plant Biology
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    • v.36 no.4
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    • pp.307-312
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    • 1993
  • In germinating pea, contents of enodgenous polyamine in the leaf and stem were determined, and protein content, peroxidase activity and pattern of isozymes were examined in the leaf treated with polyamines. During growth of the pea for 14 days in light condition, the polyamines in leaf and stem showed the highest level at the 5th day, and were decreased rapidly at the 7th day, kept almost constant level since then. The putrescine level was relatively higher than those of spermidine and spermine, and cadaverine was also detected. On the other hand, in the leaf treated with spermine (0.01 mM) protein content increased about 250% than that of the control, the peroxidase activity increased ore than 100% in spermine of 0.01 mM and 0.1 mM. In treating with putrescine of 0.1 mM the pattern of peroxidase isozyme appeared 4 new cathodic bands (pI 4.8, 5.6, 5.9 and 6.8) compared with the control, the clear cathodic bands (pI 5.6, 5.9, 6.4 and 6.6) were also observed in spermine of 0.1 mM. These results suggest that polyamines were important factor in the differentiation of pea at the early stage of germination.

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Genetic Relationships of Silkworm Stocks in Korea Inferred from Isozyme Analyses (동위효소 다형특성에 의한 누에 품종의 유연관계)

  • 성수일
    • Journal of Sericultural and Entomological Science
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    • v.39 no.2
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    • pp.119-133
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    • 1997
  • Isozyme was used to characterize general protein patterns of genetic relationships among 303 silkworm stocks preserved in National Sericultural and Entomology Research Institute, RDA. Six isozymes (esterase, acid phosphatase, alkaline phosphatase, amylase, glucose-6-phosphate dehydrogenase and sucrase) from hemolymph, midgut, and digestive juice were employed to construct dendograms(UPGMA method) using a polycrylamide gel electrophoresis. A cluster analysis revealed four major group, which were divided into several subgroups within each group, contained assemglages of Japanese and Chinese races. Especially, genetic differentiation in the first and second group was greatest rather than within Japanese and Chinese races repectively and was concordant with the hypothesis of phyletic sorting of initial variability in China many years ago. Hypothesized recent introgression between groups was also plausible, but the eviednce suggested bidirectional gene flow between the Chinese and the Japnaese lineages. Interpreting the results in light of evidence from the current study, the genetic diversity and relationship showed in Korean silkworm race, Hansammyun reflected early and independent evolution from the Chinese ancestor, limited addition of new variability and phyletic sorting within Korean peninsula more than 4,000 years.

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Evaluation and Selection of Potential Parents Based on Selection Indices and Isozyme Variability in Silkworm, Bombyx mori, L.

  • Moorthy S.M.;Das S.K.;Rao, P.R.T.;Urs S. Rao,;Sarkar A.
    • International Journal of Industrial Entomology and Biomaterials
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    • v.14 no.1
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    • pp.1-7
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    • 2007
  • In order to find out the appropriate parents for the breeding programme, twelve bivoltine and three multivoltine silkworm breeds were evaluated on the basis of multivariate selection index and isozyme analysis. Of which, four [CSR2, D6 (P), SK3, SK4] bivoltine and two multivoltine (Nistari, Cambodge) breeds were selected and breeding initiated to develop higher survival bivoltine silkworm breed suitable for tropical conditions. Among two isozyme (Esterase and acid phosphatase) analyzed, only esterase exhibited polymorphism among the bivoltine breeds. No polymorphism was observed among multivoltine in respect of esterase as well as acid phosphatase.

Comparative Study of Lactic Dehydrogenase Isozyme patterns in Clonal Derivatives of Chinese and Armenian Hamsters (Chinese hamster와 Armenian hamster에서 얻은 여러 細胞系에 대한 乳酸데히드로게나제 아이소자임 패턴에 관한 比較硏究)

  • Kang, Yung-Sun;Sahsook Hahn
    • The Korean Journal of Zoology
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    • v.17 no.1
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    • pp.37-42
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    • 1974
  • Comparative LDH isozyme studies on normal, malignant and virus-transformed derivatives of Chinese and Armenian hamsters were illustrated. With respect to LDH isozyme patterns, there were good indications that epithelial-like cells have varying levels of LDH-1-2-3 and -4, in addition to the "standard" LDH-5 band, which was exhibited by the majority of fibroblast-like cell lines.ell lines.

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The Detection of Superoxide Dismutase Activity and Isozyme Pattern of Panax ginseng C.A. Meyer Leaves (인삼엽에서 Superoxide Dismutase Activity 측정 및 Isozyme Pattern 검정)

  • Yang, Deok-Jo;Kim, Myeong-Sik;Lee, Seong-Jong
    • Journal of Ginseng Research
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    • v.11 no.1
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    • pp.24-31
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    • 1987
  • We studied a assay method on the measurement of superoxide dismutase (SOD Superoxide : superoxide oxidoreductase, EC. 1. 15. 1. 1) activity with photoreduced flavin and nitroblue tetrazolium (NBT) as superoxide (${O_2}^{-}$) source and detector, respectively. The $\Delta$E (1000 ng SOD$.$$min.)^{-1}$ of photoreduced flavin-NBT system was 0.08, whereas that of xanthine-xanthine-cytochrome system used broadly in experiments was 0.014. Therefore, the new method was regarded more simple and utilizable than xanthine-xanthine cytochrome system method. In the present paper, we also carried out to investigate the SOD activity and isozyme pattern for the parpose of study of leaf-burning disease in ginseng (Panax ginseng C.A. Meyer) leaves.

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Changes of Esterase Isozymes During the Development from Plodia interpunctella (Hiibner) (화랑곡나방의 발생에 따른 Esterase Isozymes의 Pattern변화)

  • Park, Hee-Yun;Lee, Hyung-Chul;Yoo, Chong-Myung
    • Journal of the Korean Society of Tobacco Science
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    • v.20 no.1
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    • pp.80-86
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    • 1998
  • Changes in activity and classification of esterase isozymes during the tire cycle or Plodia inteipunctella (Hiibner) were investigated by the native polyacrylamide gel electrophoresis. The stage specificity in esterase activity and isozyme pattern was observed throughout the larvalpupal-adult transformation. The activity esterase was highest at the 3-day old adult stage, and the lowest level at the prepupal stage. A total of 12 esterase bands were identified throughout the development, and the bands showing high enzyme activity was observed in the middle part of gel. Twelve esterases on the basis of inhibition by the three types of inhibitors (organophosphates, eserine sulfate and sulfhydryl reagents) were classified into three class, namely, carboxylesterase (CE), arylesterase (ArE) and cholinesterase (ChE), and these classes contained 7, 3 and 2 isozymes, respectively.

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Preliminary Studies on the Role of ${\alpha}-Esterase$ Isozymes in Quantitative Traits of Two Multivoltine Silkworm (Bombyx mori L.) Races and their $F_1$ Hybrid

  • Kumaresan P.;Somasundaram P.;Kumar K. Ashok;Urs S. Raje
    • International Journal of Industrial Entomology and Biomaterials
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    • v.13 no.2
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    • pp.113-117
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    • 2006
  • Heterosis was studied involving two multivoltine silkworm breeds viz, APM1 and SLKSPM through rearing and isozyme analysis. A positive significant heterotic effect was observed in fecundity, hatching % and survivability. The heterobeltiosis was observed only in fecundity and hatching %. Isozyme analysis of ${\alpha}-esterase$ showed variation in loci and allelic expression. The allele with heterozygosity $(Est-2^{12})$ was observed at the Est-2 locus in $F_1$ progeny. Est-3 was observed in $F_1$ progeny, whereas it was completely absent in both parental lines. The present study suggests that the markers ($Est-2^{12}$ and Est-3) targeted for introgression may be useful for the improvement of fecundity and survivability as the phenomenon of heterosis was observed only in $F_1$ progeny.

The change of ascorbate oxidase isozyme pattern during mycelial development of streptomyces lavendulae (Streptomyces lavendulae의 균사체 발달에 따른 ascorbate oxidase 동위효소 양상 변화)

  • 이현무;김재헌
    • Korean Journal of Microbiology
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    • v.26 no.2
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    • pp.137-142
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    • 1988
  • pH decreased as the substrate mycelium developed, $\Delta$pH was 1.05-1.15, but increased after the aerial mycelium formation. The lactic acid content in culture solution showed no difference between 0.2% and 5% glucose, at which the aerial mycelium formation was repressed. The growth and development of mycelium was delayed by the lactate treatment. The activity of catalase was maximum in 24 hours after inoculation, and the wuperoxide dismutase activity showed a constant level during the developmental phases. The ascorbic acid accumulated after the aerial mycelium formation. The ascorbate oxidase isozyme of Rf 0.44 appeared, while the isozyme of Rf 0.36 desappeared during the development.

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뱀장어속 어류 2종의 동위효소 및 mtDNA 분석

  • 민미숙;양서영
    • The Korean Journal of Zoology
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    • v.36 no.4
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    • pp.545-555
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    • 1993
  • 뱀장어속(Anguilla)의 뱀장어(A. ioponica)와 무태장어(A. marmoruta) 및 꾀붕장어(Anago anogo)의 유전적 특징과 종간 유연관계를 밝히기 위하여 isozyme 분석과 mtDNA 분석을 실시하였다. Isozyme 분석결과 20개의 효소 및 비효소단백질에서 총 39개의 유전자를 검출하였고 각종 특유의 genetic marker를 확인하였다. 3종의 평균 유전적 변이는 뱀장어가 HD=0.057. HG=0.065, 무태장어는 HD=0.067 HG=0.053, 체불장어가 HD=0.018. HG=0.020으로 각각 나타났다. 뱀장어와 무태장어의 평균 종간유전적 근인관계는 S=0.420 (D=0.869)로 멀게 나타났다. 6 base를 인지하는 10종류의 제한효소를 처리한 결과 mtDNA 절편양상은 각 종내에서 동일하게 나타났으나 각종간에 차이를 보였고, 종간 평균염기 치 환율은 뱀장어와 무태장어가 p=3.4%, 뱀장어속 2종과 꾀붕장어는 p=9.6%로 나타났다.

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