• Title/Summary/Keyword: isozymes

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Genic Vadadon and Speciation of Fishes of the Genus Moroco(Cyprinidae) (버들치속(잉어과) 어류의 유전적 변이 및 종분화)

  • 양서영;민미숙
    • The Korean Journal of Zoology
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    • v.32 no.2
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    • pp.75-83
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    • 1989
  • Surveys of electrophoretic variation in isozymes and general proteins encoded by 26 loci were conducted to assess species recognition and to estimate the degree of genic variation and species divergence for seven species of the genus Moroco inhabiting in Korea and Japan. Estimates of the average calculated heterozygosity per species of M semotilus, M sp., M percnurus, M lagowskii, M oxycephalus, M steindachneri and M jouyf are low: 0.021, 0.019, 0.051, 0.031, 0.023, 0.046, and 0.007, respectively, and observed heterozygosities are 0.038, 0.022, 0.060, 0.027, 0.025, 0.042, and 0.002, respectively. Allozyme analyses show these species to be distinct genetically with the lafter four species being more closely related one another than any one of them is to the rest of the species. However, these four species (M. lagowskii, M. oxycephalus, M. steindachneri and M jouyi), had unique genetic markers in each species to be recognized as valid species. These results contrast to the previous report of Chung et of. (1986) mainly due to their error in analyzing the isozyme pallems, particularly in MDH and PGI analyses. The genetic distances among M semotilus, M sp., and M percnurus are near the high end of the scale of such estimate for freshwater fish congeners. Based on estimated divergent time of these species of the genus Moroco (5 to 0.6 million years) it is assumed that they are speciated during late Pliocene to middle Pleistocene epoch prior to migration to Korean and Japanese waters through Paleo Amur River system.

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Evaluation of Mutation Rate by Differences of Isozyme Band Patterns on $M_2$ Seedling Treated with Chemical Mutagen in Barley (보리 화학돌연변이제 처리 $M_2$ 유묘의 Isozyme band pattern 차이에 의한 돌연변이율 검정)

  • Bon Cheol, Koo;M., Kucharska
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.42 no.2
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    • pp.214-219
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    • 1997
  • Mutation rate of M$_2$ plants that were treated with three types of double treatments of chemical mutagens(1.5mol Na$N_2$ + 0.75mol MNH, 0.75mol MNH + 0.75mol MNH and 0.5mol MNH + 0.5mol MNH) were estimated on the rate of chlorophyll mutant, changes of isozyme loci ; esterase (Est), glutamate oxaloacetate transaminase(GOT ; AAT) and leucyl aminopeptydase(LAP ; AMP). Rate of chlorophyll mutants (3.3% =no. of seedling carrying mutant / all number of M$_2$ seedlings $\times$ 100) and rate of esterase isozyme loci mutants(3.5% =no. of plant carrying mutant / all number of M$_2$ plant) in Dema were higher than one of Sacheon 6, but no significant differences in GOT, LAP. Among isozymes, most of mutants in M$_2$ plant of two varieties were found in esterase (73% of total mutants were occurred in esterase loci). Although many of null bands were found in GOT 3, these were not repeatable and no real mutants. It might be due to qualities of starch, amount of extract buffer and degradation of isozyme during electrophoresis and staining.

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Purification and Characterization of the External Invertase Constitutively Produced by Rhodotorula glutinis K-24 (Rhodotorula glutinis K-24에 의해 구성적으로 생산되는 세포외 Invertase의 정제 및 특성)

  • Choi, Mi-Jung;Kim, Chul;Lee, Sang-Ok;Lee, Tae-Ho
    • Microbiology and Biotechnology Letters
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    • v.18 no.4
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    • pp.368-375
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    • 1990
  • Rhodoto& ghtbth~ K-24 was found to produce external invertase in addition to internal and cell wall bound invertase. External invertase was purified to an electrophoretically homogeneous state and partitally characterized and was compared with internal and cell wall bound invertase of which procedures for purification and characterization were reported previously. The enzyme was purified by ethanol precipitation, column chromatographies on DEAE-Sephadex A-50 and SP-Sephadex C-50, and gel filtration on Sephadex G-100. The molecular weight and subunit molecular weight of external invertasGwere estimated to be 220,000 and 100,000, respectively. The isoelectric point of the enzyme was about pH 6.0. The optimum pH and temperature for enzyme activity were pH 4.0 and $60^{\circ}C$, respectively. The enzyme remained stable at the wide range, from pH 3.0 to 11.0 and stable up to $40^{\circ}C$, but was inactivated at temperatures above that. $HgC_12, AgN0_3, MnS0_4$, SDS and p-CMB inhibited the enzyme activity. The $K_m$ value of the enzyme for sucrose was $1.0\times 10^{-2}$M. From these results, the three isozymes from Rh. glutinis K-24 seem to have the similar enzymatic properties, but to differ in molecular and subunit weights.

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Phylogeny of Bombyx mandarina inhabiting Korea analysing the isozyme and hemolymph protein polymorphism (동위효소와 체액단백질 분석에 의한 한국산 멧누에나방의 지역적 특성)

  • 이재만;김경아;노시갑
    • Journal of Sericultural and Entomological Science
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    • v.45 no.1
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    • pp.18-24
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    • 2003
  • B. mandarina of Korean population apparently differs B. mori in isozyme analysis. Fourteen polymorphism occurred B. mandarina not in B. mori at 6 isozymes, Bph, Bes, Amy-hc, Ies, Ict-D, Ict-E. Korean population has shared with the Korean native strain of B. mori in B genotype of Bes, F of Amy-hc, n of Ict-E, M and S of Ict-H. These 5 genotype were known that detection only Korean native strains of B. mori. Nei's genetic distance based on the genotype of isozyme and hemolymph protein using 4 populations of B. mandarina varied from 0.0350 to 0.0624. The distances of 0.0350 is between Jinju and Chilgok population and between Jinju and Kosung population has the largest distances, 0.0624. In genus of Bombyx, B. mandarina and B. mori, genetic distance varied from 0.3822 to 0.5074. Phylogenetic tree obtained using the subprogram UPGMA of NTSYS represented that Bombyx devided two group, B. mandarina and B. mori. B. mandarina has genetic differences according to the population within the Korean peninsula, but that was not recognized genetic variation or divergence considering low values of genetic distance.

Localization and isozyme patterns of phosphatase in Fibricola seoulensis (Fibricola seoulensis에서 phosphatase의 분포와 동위효소유형)

  • 김홍자;김창환
    • Parasites, Hosts and Diseases
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    • v.31 no.4
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    • pp.353-362
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    • 1993
  • The present study was carried out to investigate the localization and isozyme patterns of acid phosphatase and alkaline phosphatase in metacercariae and in adults of F. seoulensis by enzyme-histochemistry method and electrophoresis. Acidphosphatase showed a strong activity at pH 5 in the intestinal caecum of adults, but showed no reactions in the nonsubstrate control and in the inhibitor-treated control. Alkaline phosphatase showed a strong activity at pH 8 in the intestinal caecum and the tribocytic organ of adults, and in the intestinal caecum and in the genital anlagen of metacercariae. In non-denature PAGE, ten bands of protein fraction from the extracts of metacercariae and twenty-two bands from adults were detected. In denature PAGE, two protein bands having molecular weights of 192 kDa and 123 kDa were detected in the metacercariae, but absent from adult stage. In adults, protein fractions of 27.5 kDa, 24.5 kDa, 21.4 kDa, 18 kDa, 16 kDa and 15 kDa were detected. In non-denature PAGE, isozymes of acid phosphatase showed the most strong activity at pH 5, whereas no activity was shown at pH 2 and pH 7. One isozyme 85 kDa, 73 kDa and 62 kDa) in adults.

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Proteomic Analysis for Neuroprotective Effect of Gastrodia elata Blume in the Substantia Nigra of Mice (천마의 흑질 내 도파민성 신경세포 보호 효과에 대한 단백체학적 분석)

  • Chang-Hwan, Bae;Hee-Young, Kim;Hanul, Lee;Ji Eun, Seo;Dong Hak, Yoon;Seungtae, Kim
    • Korean Journal of Acupuncture
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    • v.39 no.4
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    • pp.142-151
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    • 2022
  • Objectives : Parkinson's disease (PD) is a neurodegenerative disorder threatening the quality of life and highly occurred in over 65 years old. Gastrodia elata Blume (GEB), a traditional medicine used for the treatment of headache and convulsion, has been reported to have neuroprotective effect. This study was designed to investigate the neuroprotective effect of GEB and the proteomic changes in the substantia nigra (SN) of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-treated mice. Methods : Male eleven-week-old C57BL/6 mice were intraperitoneally injected with 30 mg/kg of MPTP at 24-h intervals for 5 days. Two hours after the daily MPTP injection, the mice were orally administered 800 mg/kg of GEB extract, which continued for 7 days beyond the MPTP injections, for a total of 12 consecutive days. Two hours after the final GEB administration, the brain samples were collected, and dopaminergic neuronal death and proteomic changes in the SN were evaluated. Results : GEB prevented the MPTP-induced dopaminergic neuronal death and regulated the expression of 11 proteins including thimet oligopeptidase, T-complex protein 1, glycine tRNA ligase, and pyruvate kinase isozymes M1. Conclusions : GEB prevents MPTP-induced dopaminergic neuronal death by regulating the proteins in the SN.

Appropriate Electrophoresis Techniques and Isozymes to Identification of Barley Cultivars (보리품종 구분에 적합한 전기영동법과 효소)

  • Son, Eung-Ryong;Lee, Yong-Se;Yoon, Kyung-Eun;Ha, Yong-Woong
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.30 no.4
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    • pp.405-411
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    • 1985
  • The buffer soluble proteins were extracted from six cultivars of barley grains and analyzed by various electrophoresis; 7.5% polyacrylamide slab gel, 2-30% polyacrylamide porosity gradient tube gel, isoelectric focusing (pH4-9) and starch gel electrophoresis. The proteins, esterase, acid phosphatase, malate dehydrogenase, glutamate dehydrogenase and leucine aminopeptidase were investigated to find out the best method to differentiate barley cultivars. The result were that protein and esterase bands in 2-30% polyacrylamide porosity gradient tube gel electrophoresis and protein bands in 7.5% polyacrylamide slab gel electrophoresis showed typical varietal differences. Therefore, those methods were suitable for differentiation of barley cultivars. It was difficult to differentiate the cultivars by the other methodes and patterns of the other enzymes.

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Development and Utilization of KASP Markers Targeting the Lipoxygenase Gene in Soybean

  • Seo-Young Shin;Se-Hee Kang;Byeong Hee Kang;Sreeparna Chowdhury;Won-Ho Lee;Jeong-Dong Lee;Sungwoo Lee;Yu-Mi Choi;Bo-Keun Ha
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.68 no.4
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    • pp.294-303
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    • 2023
  • Lipoxygenase gives soybeans their grassy flavor, which can disrupt food processing efficiency. This study aimed to identify soybean genotypes with lipoxygenase deficiency among 1,001 soybean accessions and to develop kompetitive allele specific PCR (KASP) markers that can detect lipoxygenase mutations. Three lipoxygenase isozymes (Lox1, Lox2, and Lox3) were analyzed using a colorimetric assay based on a substrate-enzyme reaction. Among the 1,001 accessions examined, two (IT160160 and IT276392) exhibited a deficiency solely in Lox1, and one (IT269984) lacked both Lox1 and Lox2. IT160160 had a 74-bp deletion in exon 8 of Lox1 (Glyma13g347600), whereas IT276392 displayed a missense mutation involving the change of C to A at position 2,880 of Lox1. Moreover, we successfully developed four KASP markers that specifically target Lox1, Lox2, and Lox3 mutations. To validate the Lox1 KASP markers, we used two F2:3 populations generated through a cross between Daepung 2 (lipoxygenase wild type, maternal parent), IT160160, and IT276392 (null Lox1, paternal parent). The results revealed that the Daepung 2 × IT160160 group followed the expected 3:1 ratio according to Mendel's law, whereas the Daepung 2 × IT276392 group did not. Furthermore, a comparison between the colorimetric and KASP marker analyses results revealed a high agreement rate of 96%. KASP markers offer a distinct advantage by allowing the distinction of heterozygous types independent of other variables. As a result, we present an opportunity to expedite the lipoxygenase-deficient cultivar development.

Difference in Electrophoretic Phenotypes of Rice Cultivars Selected to Oxyfluorfen (Oxyfluorfen에 대한 내성(耐性) 및 감수성(感受性) 수도품종(水稻品種)의 전기영동(電氣泳動) 표현형(表現型) 차이(差異))

  • Kuk, Y.I;Guh, J.O.;Lee, D.J.;Kim, Y.J.
    • Korean Journal of Weed Science
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    • v.8 no.2
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    • pp.199-207
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    • 1988
  • The study was intended to know any relations between the rice tolerance to oxyfluorfen and varietal speciation in seed protein composition or any enzymatical allelies with or without chemical treatment. Rice varieties used were Chokoto, Aichiasahi, Agabyeo, IR 3941 and Tablei as the tolerant group, and Mushakdanti, Weld Pally, HP 1033, HP 857, and HP 907 as the susceptible, respectively. Electrophoretic methods used were SDS-PAGE for seed protein, 7% PAGE for isozymes (acid phosphatase and peroxidase from rice seedling) and changes in isoenzyme activity (malate dehydrogenase, peroxidase and esterase) as affected by oxyfluorfen treatment ($10^{-4}M$) was also studied. The results are summarized as follows. -Among 19 bands separated in seed proteins, two different rice groups selected in terms of tolerance were clustered in dissimilarity. This was based on 2 facts in that G band was not present in susceptible varieties and that less activity of H, N, O, P, Q, Rand S band was shown. -Among 4 bands separated in acid phosphatase, the presence of (band and lower activity of B band was specific for tolerant varieties. For 4 minor bands separated in peroxidase, the tolerant varieties had no activity in B band and higher activity in A, C, D bands. -Time-course study of isozymes as affected by $10^{-4}M$ oxyfluorfen showed that Chokoto, the tolerant varieties, had little activity in A band and consistently higher activities in Band C bands for malate dehydrogenase. For 5 bands separated in peroxidase, B band was not found in Chokoto while A, C, D, and E bands were consistently present. Esterase was separated into about 4 bands in which Chokoto had maintained higher activities in A, C and D bands.

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Investigation of Herbicide Safeners and their Mode of Safening Action;II. Effect of N-(4-chlorophenyl) maleimide, Plant Growth Regulators, and Alkylating Agents on Glutathione Content and Glutathione S-transferase Activity (제초제(除草劑) 약해경감물질(藥害輕減物質) 탐색(探索)과 작용기구(作用機構) 규명(糾明);Ⅱ. Glutathione 함량(含量)과 Glutathione S-transferase 활성(活性) 변화(變化)에 대한 N-(4-chlorophenyl) maleimide, 식물생장조절물질(植物生長調節物質) 및 Alkylating Agents 의 효과(效果))

  • Chun, Jae-Chul;Ma, Sang-Yong
    • Korean Journal of Environmental Agriculture
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    • v.14 no.3
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    • pp.329-337
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    • 1995
  • The effect of N-(4-chlorophenyl) maleimide(CPMI), plant growth regulators, and alkylating agents on gluathione(GSH) content and glutathione S-transferase(GST) activity was examined with 3-day-old etiolated sorghum(Sorghum bicolor [L.] Moench) seedlings. The GSH content and GST activity of untreated seedlings were higher in shoots than that in roots. Response of GST activity in coleoptile was significantly greater than in other tissues of sorghum seedling. In CPMI-treated seedlings, GSH content was not significantly different from that in untreated seedlings. CPM treatment resulted in 2.3-fold increase in GST activity measured with metolachlor as substrate in the coleoptile region. In contrast, change in GST activity measured with metolachlor as substrate in the coleoptile region. In contrast, change in GST activity measured with 1-chloro-2, 4-dinitrobenzene did not occur. The increase of GST activity was caused by induction of a GST isozyme, which is substrate-specific to metolachlor. Subsequently, two hypotheses related to metolachlor detoxification were evaluated on the basis of regulation of plant growth regulators and substrate induction of GST activity. In coleoptile, GST activity measured with metolachior was increased to 2.1-and 3.4-fold by both 2, 4-dichlorophenoxyacetic acid(2,4-D) and metolachlor treated at the germination stage of sorghum, respectively. Treatments of 2.4-D and metolachlor also induced isozymes exhibiting the activity toward metolachlor. One of the isozymes was co-eluted with that induced by CPMI. These results indicated that increase in GST activity by CPMI may be partially related to auxin regulation and substrate induction.

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