• Title/Summary/Keyword: Isozyme analysis

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Presence of Carbonic Anhydrase III-like Protein in Shaggy Sea Raven, Hemitripterus villosus (삼세기(Shaggy sea raven, Hemitripterus villosus)의 carbonic anhydrase III에 관한 연구)

  • Kweon, Rok Eun;Kho, Kang Hee
    • Journal of Life Science
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    • v.24 no.2
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    • pp.186-190
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    • 2014
  • Carbonic anhydrase isozymes (CAs) are widespread zinc-containing metalloenzyme family. The enzyme catalyzes the reversible interconversion of $CO_2$ and $HCO_3$. This reaction is the main role of CA enzymes in physiological conditions. CA III, one of the CA isozymes, has been identified in many tissues. It is distinguished from the other isozymes by several characteristics, particularly by a lower specific activity and by its resistance to acetazolamide. However, the physiological function of CA III in fish is unknown. In this study, we examined the detection of CAs in the Shaggy sea raven Hemitripterus villosus, using SDS-PAGE, isoelectric focusing (IEF), and western blot analysis. We detected a significant protein band with molecular weight about 30 kDa from the tissues of H. villosus by SDS-PAGE and western blotting. A specific band of CA III with pI 7.0 was detected by IEF and western blotting in gill and muscle. The immunoreaction of anti-CA III expressed in the gill of H. villosus was much stronger than other tissues. One explanation for this result is that the fish gill is the only organ that is exposed to the external environment and that plays an important role in acid-base relevant ion transfer, the transfer of $H^+$ and/or $HCO{_3}^-$, for the maintenance of systemic pH. This is the first report on the identification of a carbonic anhydrase III-like protein from H. villosus.

Effects of deoxynivalenol- and zearalenone-contaminated feed on the gene expression profiles in the kidneys of piglets

  • Reddy, Kondreddy Eswar;Lee, Woong;Jeong, Jin young;Lee, Yookyung;Lee, Hyun-Jeong;Kim, Min Seok;Kim, Dong-Woon;Yu, Dongjo;Cho, Ara;Oh, Young Kyoon;Lee, Sung Dae
    • Asian-Australasian Journal of Animal Sciences
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    • v.31 no.1
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    • pp.138-148
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    • 2018
  • Objective: Fusarium mycotoxins deoxynivalenol (DON) and zearalenone (ZEN), common contaminants in the feed of farm animals, cause immune function impairment and organ inflammation. Consequently, the main objective of this study was to elucidate DON and ZEN effects on the mRNA expression of pro-inflammatory cytokines and other immune related genes in the kidneys of piglets. Methods: Fifteen 6-week-old piglets were randomly assigned to three dietary treatments for 4 weeks: control diet, and diets contaminated with either 8 mg DON/kg feed or 0.8 mg ZEN/kg feed. Kidney samples were collected after treatment, and RNA-seq was used to investigate the effects on immune-related genes and gene networks. Results: A total of 186 differentially expressed genes (DEGs) were screened (120 upregulated and 66 downregulated). Gene ontology analysis revealed that the immune response, and cellular and metabolic processes were significantly controlled by these DEGs. The inflammatory stimulation might be an effect of the following enriched Kyoto encyclopedia of genes and genomes pathway analysis found related to immune and disease responses: cytokine-cytokine receptor interaction, chemokine signaling pathway, toll-like receptor signaling pathway, systemic lupus erythematosus (SLE), tuberculosis, Epstein-Barr virus infection, and chemical carcinogenesis. The effects of DON and ZEN on genome-wide expression were assessed, and it was found that the DEGs associated with inflammatory cytokines (interleukin 10 receptor, beta, chemokine [C-X-C motif] ligand 9, CXCL10, chemokine [C-C motif] ligand 4), proliferation (insulin like growth factor binding protein 4, IgG heavy chain, receptor-type tyrosine-protein phosphatase C, cytochrome P450 1A1, ATP-binding cassette sub-family 8), and other immune response networks (lysozyme, complement component 4 binding protein alpha, oligoadenylate synthetase 2, signaling lymphocytic activation molecule-9, ${\alpha}$-aminoadipic semialdehyde dehydrogenase, Ig lambda chain c region, pyruvate dehydrogenase kinase, isozyme 4, carboxylesterase 1), were suppressed by DON and ZEN. Conclusion: In summary, our results indicate that high concentrations of DON and ZEN suppress the inflammatory response in kidneys, leading to potential effects on immune homeostasis.

Association of Single Nucleotide Polymorphism (SNP) in the PGK 2 Gene with Growth Traits in Pigs (돼지 PGK 2 유전자의 단일염기다형성 및 성장 형질과의 연관성 구명)

  • Jang, Hong-Chul;Kim, Sang-Wook;Lim, Da-Jeong;Kim, Jae-Young;Cho, Kyu-Ho;Kim, Myung-Jick;Lee, Ji-Woong;Choi, Bong-Hwan;Kim, Tae-Hun
    • Journal of Animal Science and Technology
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    • v.53 no.1
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    • pp.15-22
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    • 2011
  • The purpose of this study was to analyse of association between growth traits and single nucleotide polymorphisms (SNPs) polymorphism of phosphoglycerate kinase 2 (PGK 2) gene in pigs. The birth weight of piglet influences on weaning weight and survival rate that are import economic traits in pig industry. Also, these growth traits are representative factor to decrease a period getting to marketing weight as well as growth rate in pig. The PGK 2 is an isozyme that catalyzes the first ATP-generating step in the glycolytic pathwayand important enzyme related with energy metabolism. Twenty of SNPs were discoveredby genome structure analysis that compares the sequence on promoter and transcription region of PGK 2 gene in porcine chromosome 7. An association between PGK 2 SNPs and growth traits was analyzed in $F_2$ reciprocal-crossbred population between korean native pig (KNP) and Landrace. Association analysis indicated that polymorphism of the PGK 2 gene promoter region has significant effects on weight at birth (p<0.01) and weight at 3 weeks of age (p<0.0001). These results suggest that PGK 2 gene polymorphism was associated with energy metabolism and physiological function of growth in pig.

GEOMETRIC NINLINEAR ANALYSIS OF UNERGROUND LAMINATED COMPISITE PIPES (기하학적 비선형을 고려한 지하매설 복합재료 파이프의 해석)

  • 김덕현;이인원;변문주
    • Computational Structural Engineering
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    • v.2 no.1
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    • pp.65-70
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    • 1989
  • An analytical study was conducted using the Galerkin technique to determine behaviour of thin fibrereinforced and laminated composite pipes under soil pressure. Geometric nonlinearity and material linearity have been assumed. It is assumed that vertical and lateral soil pressure are proportional to the depth and lateral displacement of the pipe respectively. It is also assumed that radial shear stress is negligible because the ratio of thickness to the radius of pipe is very small. The above results are verified by the finite element analysis.

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Interorder Hybridization between Pleurotus ostreatus and Elfvingia applanata by Protoplast Fusion (원형질체(原形質體) 융합(融合)에 의한 느타리버섯과 잔나비걸상버섯의 이목간(異目間) 교잡(交雜))

  • Yoo, Young-Bok
    • The Korean Journal of Mycology
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    • v.22 no.1
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    • pp.107-116
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    • 1994
  • Interorder somatic hybrids were obtained by protoplast fusion between Pleurotus ostreatus in the order Agaricales and Elfvingia applanata in the order Aphyllophorales. The fusants were classified into stable heterokaryons and spontaneously segregated heterokaryons. Hyphae of all fusion products except two strains did not form clamp connections. Out of them, two clamped and three clampless fusants produced mature fruiting bodies by light-dark cycle on sawdust rice bran medium. All of these basidiocarps had clamp connections. Three fusants were analysed with the distribution of progenies and segregation of genetic characters by random spore analyses. The genetic markers were shown to segregate and recombine in the first generation of monospores isolated from basidiocarps. Phenotypes of a large number of auxotrophic progenies were not detected in the two clamped fusants. The aberration ratio of segregants indicated the gene interaction resulting from different genome structure between distantly related species. The polymerase chain reaction (PCR) was adopted for the detection of somatic hybrids nuclear DNA. Four fusants showed a positive results in three kinds of primers. The prominent reaction products are represented by new bands in primer # 87 and # 125. Out of four fusants, two somatic hybrids had non-parental mtDNA patterns when digested with EcoR1 and HindIII. Comparison of somatic hybrids, tissue culture isolates(TC) and multispore germination isolates(MS) were made using esterase isozyme analysis. It is apparent that somatic hybrids had a minor banding patterns which are quite different from those of parents.

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Morphological and Genetic Variation of Allium victorialis var. platyphyllum (산마늘(Allium victorialis var. platyphyllum)의 형태적.유전적 변이)

  • Bae, Kwan Ho;Hong, Sung Cheon
    • Current Research on Agriculture and Life Sciences
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    • v.13
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    • pp.45-53
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    • 1995
  • This research was conducted to investigate morphological and genetic variation of Allium victorialis val. platyphyllum which growed wild in Mt. Hambaek, Mt. Odae, and Ullungdo. The tree layer of Allium victorialis var. platyphyllum community in Mt. Hambaek and Mt. Odae was dominated by Quercus mongolica. The tree layer of Ullungdo generally consist of Fagus crenata var. multinervis, Acer triflorum, Sorbus commixta, and Tilia insularis. In the herb layer, Rumohra standishii, Trillium tschonoskii, and Lilium hansonii are common at Allium victorialis var. platyphyllum community in Ullungdo. The vegetation in Ullungdo was widely different from those in Mt. Hambaek and Mt. Odae by species composition. The result of Principal Component Analysis(PCA) and Canonical Discriminent Analysis of by the 8 characters showed that Allium victorialis var. platyphyllum could be classified into 2 groups: (one ; Mt. Hambaek and Mt. Odae, the other ; Ullungdo). In PCA, the major factors in the first principal component group was angle of leaf apex. Variation of band by isozyme GOT(glautamate oxaloaccetate transaminase) is similar between Mt. Hambaek and Mt. Odae. However, Ullungdo differed from Mt. Hambaek and Mt. Odae in variation of bands.

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Genetic Variation of Pinus densiflora Populations in South Korea Based on ESTP Markers (ESTP 표지를 이용한 국내 소나무 집단의 유전변이)

  • Ahn, Ji Young;Hong, Kyung Nak;Lee, Jei Wan;Hong, Yong Pyo;Kang, Hoduck
    • Korean Journal of Plant Resources
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    • v.28 no.2
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    • pp.279-289
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    • 2015
  • Genetic diversity and genetic differentiation of thirteen Pinus densiflora populations in South Korea were estimated using nine ESTP (Expressed Sequence Tag Polymorphism) markers. The numbers of allele and the effective allele were 2.2 and 1.8, respectively. The percentage of polymorphic loci (P) was 98.8%. The observed and the expected heterozygosity were 0.391 and 0.402, respectively, and the eleven populations except for Ahngang and Gangneung population were under Hardy-Weinberg equilibrium state. The level of genetic differentiation (Wright’s FST = 0.057) was higher than those of isozyme or nSSR markers. We could not find out any relationship between the genetic distance and geographic distribution among populations from cluster analysis. Also, the genetic differentiation between populations was not correlated with the geographic distance (r = 0.017 and P = 0.344 from Mantel test). From the result of FST-outlier analysis to identify a locus under selection, six loci were detected at confidence interval of 99% by the frequentist’s method. However, only three loci (sams2+AluⅠ, sams2+RsaⅠ, PtNCS_p14A9+HaeⅢ) were presumed as outliers by Bayesian method. The sams2+AluⅠ and sams2+RsaⅠlocus were originated from the sams2 gene and seemed to be the loci under balancing selection.

Physiological Roles of Phospholipase Cγ and Its Mutations in Human Disease (Phospholipase Cγ의 생리적 기능과 질병과 연관된 돌연변이)

  • Jang, Hyun-Jun;Choi, Jang Hyun;Chang, Jong-Soo
    • Journal of Life Science
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    • v.30 no.9
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    • pp.826-833
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    • 2020
  • Phospholipase C gamma (PLCγ) has critical roles in receptor tyrosine kinase- and non-receptor tyrosine kinase-mediated cellular signaling relating to the hydrolysis of phosphatidylinositol 4,5-bisphosphate [PI(4,5)P2] to produce inositol 1,4,5 trisphosphate (IP3) and diacylglycerol (DAG), which promote protein kinase C (PKC) and Ca2+ signaling to their downstream cellular targets. PLCγ has two isozymes called PLCγ1 and PLCγ2, which control cell growth and differentiation. In addition to catalytically active X- and Y-domains, both isotypes contain two Src homology 2 (SH2) domains and an SH3 domain for protein-protein interaction when the cells are activated by ligand stimulation. PLCγ also contains two pleckstrin homology (PH) domains for membrane-associated phosphoinositide binding and protein-protein interactions. While PLCγ1 is widely expressed and appears to regulate intracellular signaling in many tissues, PLCγ2 expression is restricted to cells of hematopoietic systems and seems to play a role in the regulation of immune response. A distinct mechanism for PLCγ activation is linked to an increase in phosphorylation of specific tyrosine residue, Y783. Recent studies have demonstrated that PLCγ mutations are closely related to cancer, immune disease, and brain disorders. Our review focused on the physiological roles of PLCγ by means of its structure and enzyme activity and the pathological functions of PLCγ via mutational analysis obtained from various human diseases and PLCγ knockout mice.

Effects of Alcohol on Neurocognitive Function, Psychomotor Performance and Subjective Response in Koreans with Different ALDH2 Genotypes (한국인에서 Aldehyde Dehydrogenase 2 유전자 변이가 알코올의 신경인지 기능, 정신운동성 수행 및 주관적 반응에 미치는 영향)

  • Shin, Il-Seon;Yoon, Jin-Sang;Kim, Hyun;Yoon, Bo-Hyun;Lee, Hoon;Jung, Jae-Sung;Lee, Hyung-Yung
    • Korean Journal of Biological Psychiatry
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    • v.6 no.2
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    • pp.176-188
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    • 1999
  • Objective : The purpose of this study was to evaluate the effects of alcohol on neurocognitive function, psychomotor performance and subjective response in healthy Korean adults with different ALDH2 genotypes. Method : A total of 24 males, half with active $ALDH2^*1/2^*1$ and the other with inactive $ALDH2^*1/2^*2$, was selected through genotyping using restriction fragment length polymorphism. In a double-blind, placebo-controlled cross-over design, each subject consumed 0.5g/kg dose of alcohol, given as a mixture of 40% vodka and orange juice, and placebo(orange juice) on two separate occasions on an average of weekly intervals. The blood alcohol concentrations(BACs) were measured using a breath analyzer at baseline and at 30, 60 minutes after drinking. P300s were measured at baseline and at 30 minutes after alcohol and placebo intake. Vital signs and psychomotor performance[Critical Flicker Fusion Threshold(CFFT), Choice Reaction Time (CRT), Digit Symbol Substitution(DSS)] were measured at baseline and at 60 minutes after alcohol and placebo intake. Subjective responses were measured at the end of the study. The statistical analysis focused on whether there were any differences between groups with different ALDH2 genotypes. Results : The major results are as follows. 1) BACs in the inactive group were overall equivalent to those in the active group. Only in terms of time, BACs were significantly higher overall at 30 minutes than at 60 minutes after alcohol intake. 2) Pulse rates were significantly increased after alcohol intake compared with placebo, and the increase was greater in the inactive than in the active group. 3) P300 latencies in leads Fz(frontal), Cz(cental) and Pz(parietal) were significantly increased after alcohol intake compared to placebo, and the increase was greater in the inactive than in the active group. P300 amplitudes in leads Cz and Pz were significantly decreased overall after alcohol intake compared to placebo. 4) Compared with placebo, alcohol produced significant effect on the psychomotor performance : impairment in the inactive group, improvement in the active group. 5) Compared with placebo, alcohol significantly induced a negative or an intense effect on the subjective responses in the inactive group, but little negative and even a somewhat positive effect in the active group. Conclusions : These results suggest that ALDH isozyme variance might be an important factor to determine the effects of acute dose of alcohol on the various psychobehavioural functions and also to determine the alcohol use pattern and to predict the future development of alcohol overuse and/or abuse.

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Phytopathogenicity of Burkholderia gladioli pv. alliicola CH1 and Production of PGase Isozymes (Burkholderia gladioli pv. alliicola CH1의 병원성 및 Polygaractronase Isozymes 생성)

  • Lee, Chan-Jung;Lee, Jong-Tae;Kim, Yeong-Tae;Jhune, Chang-Sung;Cheong, Jong-Chun;Park, Wan
    • Research in Plant Disease
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    • v.18 no.3
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    • pp.240-244
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    • 2012
  • Burkholderia gladioli pv. alliicola CH1 showed typical soft rot symptoms at higher than $20^{\circ}C$ but very weak soft rot symptoms at temperature under $10^{\circ}C$. Among the nine agro-chemicals, oxolinic acid WP, streptomycin + copper hydroxide WP and streptomycin WP were found to be effective for the inhibition of the pathogen in vitro. The results of scanning electron microscopic investigation showed that onion bulbs was macerated by infection of B. gladioli pv. alliicola CH1. B. gladioli pv. alliicola CH1 was able to produce polygalacturonase but did not produce pectin lyase and carboxymethylcellulase. In analysis of the polygalacturonase activity of the isolated pectin-degradation enzymes from B. gladioli pv. alliicola CH1 total protein, three activity bands 45 kDa, 35 kDa, and 29 kDa were detected by the direct (or in-gel) activity staining on SDS-PAGE.