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cDNA Cloning and Overexpression of an Isoperoxidase Gene from Korean-Radish, Raphanus sativus L.

  • Park, Jong-Hoon;Kim, Soung-Soo
    • BMB Reports
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    • v.29 no.2
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    • pp.137-141
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    • 1996
  • A partial cDNA encoding a Korean radish isoperoxidase was obtained from a cDNA library prepared from 9 day old radish root. In order to obtain Korean radish isoperoxidase cDNA, 5' RACE (rapid amplification cDNA end) PCR was performed and a cDNA (prxK1) encoding a complete structural protein was obtained by RT (reverse transcription)-PCR. Sequence analysis revealed that the length of the cDNA was 945 base pairs, and that of the mRNA transcript was ca. 1.6 kb. The deduced amino acid of the protein were composed of 315 amino acid residues and the protein was 92% homologous to turnip peroxidase, and 46% to 50% homologous to other known peroxidases. The 945 bp cDNA encoding Korean radish isoperoxidase was overexpressed in Escherichia coli up to approximately 9% of total cellular protein. The recombinant fusion protein exhibited 43 kDa on SDS-PAGE analysis and the activity level of the recombinant nonglycosylated protein was two fold higher in IPTG induced cell extracts than that of uninduced ones.

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Centering Theory and Argument Deletion in Spoken Korean (센터링 이론과 대화체에서의 논항 생략 현상)

  • 홍민표
    • Korean Journal of Cognitive Science
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    • v.11 no.1
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    • pp.9-24
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    • 2000
  • This paper analyzes the distribution and classification of unrealized arguments of a predicate often called zero pronouns. in spoken Korean. Based on the transcript of a one-hour-Iong dialogue. recorded from public radio stations. I present the statistical data on argument ellipsis in Korean with respect to the frequency of zero ronouns as well as the nature of their antecedents. I go further to review some of the previous efforts to identify the discourse- theoretic functions of zero-pronouns in the framework of Centering Theory. and propose that the zero-pronouns in spoken Korean be divided into center-insensitive vs. center-sensitive classes. I also point out a couple of language-particular idiosyncrasies found in Korean, such as morpho-syntactic elements and encyclopaedic knowledge. that interact with center management in on-going discourse and often lead to difficulties in applying the centering rules and constraints to Korean.

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Arabidopsis SIZ1 positively regulates alternative respiratory bypass pathways

  • Park, Bong-Soo;Kim, Sung-Il;Song, Jong-Tae;Seo, Hak-Soo
    • BMB Reports
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    • v.45 no.6
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    • pp.342-347
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    • 2012
  • Plant mitochondria possess alternative respiratory pathways mediated by the type II NAD(P)H dehydrogenases and alternative oxidases. Here, E3 SUMO ligase was shown to regulate alternative respiratory pathways and to participate in the maintenance of carbon and nitrogen balance in Arabidopsis. The transcript abundance of the type II NAD(P)H dehydrogenases NDA2 and NDB2 and alternative oxidases AOX1a and AOX1d genes was low in siz1-2 mutants compared to that in wild-type. The addition of nitrate or ammonium resulted in a decrease or an increase in the expression of the same gene families, respectively, in both wild-type and siz1-2 mutants. The amount of free sugar (glucose, fructose and sucrose) was lower in siz1-2 mutants than that in wild-type. These results indicate that low nitrate reductase activity due to the AtSIZ1 mutation is correlated with an overall decrease in alternative respiration and with a low carbohydrate content to maintain the carbon to nitrogen ratio in siz1-2 mutants.

Cellular Signaling Molecules Associated with Peptidoglycan-Induced CCL3 Up-Regulation

  • Kim, Kang-Seung;Rhim, Byung-Yong;Eo, Seong-Kug;Kim, Koan-Hoi
    • Biomolecules & Therapeutics
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    • v.19 no.3
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    • pp.302-307
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    • 2011
  • Peptidoglycan (PGN) is detected in inflammatory cell-rich regions of human atheromatous plaques. The present study investigated the effects of PGN on CC chemokine ligand 3 (CCL3) expression, which is elevated in the atherosclerotic arteries, and determined cellular factors involved in PGN-mediated CCL3 up-regulation in mononuclear cells, with the goal of understanding the molecular mechanisms of inflammatory responses to bacterial pathogen-associated molecular patterns in diseased arteries. Exposure of human monocytic leukemia THP-1 cells to PGN resulted in enhanced secretion of CCL3 and profound induction of the CCL3 gene transcript. Both events were abrogated by oxidized 1-palmitoyl-2-arachidonosyl-sn-phosphatidylcholine, an inhibitor of Toll-like receptors 2/4. Pharmacological inhibitors such as U0126, SP6001250, Akt inhibitor IV, rapamycin, RO318220, diphenyleneiodonium chloride, and N-acetylcysteine also significantly attenuated PGN-mediated CCL3 up-regulation. However, polymyxin B, LY294002, and SB202190 did not influence CCL3 expression. We propose that PGN contributes to enhanced CCL3 expression in atherosclerotic plaques and that Toll-like receptors (TLR2), Akt, mTOR, mitogen-activated protein kinase, and reactive oxygen species are involved in that process.

Molecular cloning and expression of glyceraldehyde-3-phosphate dehydrogenase gene under environmental stresses in sweetpotato

  • Kim, Young-Hwa;Song, Young-Sun;Huh, Gyung-Hye
    • Journal of Plant Biotechnology
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    • v.35 no.2
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    • pp.95-100
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    • 2008
  • Glyceraldehyde-3-phosphate dehydrogenase (GAPDH), a main enzyme in the glycolytic pathway, is involved in cellular energy production and regarded as a housekeeping gene. Previously, cytosolic GAPDH was selected as the most significantly abundant gene in EST library of sweetpotato suspension cells. In this study, a full-length of cDNA clone (IbGAPDH) encoding GAPDH was isolated from suspension-cultured cells of sweetpotato (Ipomoea babatas), and its expression was investigated with a view to understanding the physiological function of GAPDH in relation to environmental stresses. IbGAPDH encoded a 36.9 kDa polypeptide consisting of 337 amino acids. When the deduced amino acid of IbGAPDH was compared with other higher plants, IbGAPDH showed high homology with cytosolic GAPDH. The mRNA level of IbGAPDH significantly increased under environmental stresses, such as $H_2O_2$, MV and cold treatments. Among them, the transcript level of IbGAPDH gene was the highest under cold stress. Further investigation of the transcription level under $10^{\circ}C$ or $15^{\circ}C$ was performed with different tissues of sweetpotato. The transcription of IbGAPDH was increased by cold stress with tissue-specificity, moreover, showed different patterns according to temperature.

Brassinosteroids-mediated regulation of ABI3 is involved in high-temperature induced early flowering in plants

  • Hong, Jeongeui;Sung, Jwakyung;Ryu, Hojin
    • Journal of Plant Biotechnology
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    • v.45 no.2
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    • pp.83-89
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    • 2018
  • The interplay of plant hormones is one of the essential mechanisms for plant growth and development. A recent study reported that Brassinosteroids (BR) and ABSCISIC ACID (ABA) interact antagonistically in early seedling developments through the BR-mediated epigenetic repression of ABSCISIC ACID-INSENSITIVE 3 (ABI3). However, the other physiological roles of the BR-mediated regulation of ABI3 and ABA responses beyond early seedling developments remain largely unknown. Here, we showed that the activation of BR signaling by high temperatures promotes flowering time through the suppression of ABI3 expressions. Elevated ambient temperature induced early flowering in wild type Col-0 plants, but not in BR-defective bri1-116 mutant plants. Conversely, a hyper BR biosynthetic dwf4-D mutant displayed more sensitive thermomorphic long shoot elongation and early flowering. Both expression patterns and physiological responses supported the biological roles of ABI3 in the regulation of floral transition and reproduction under high temperature conditions. Finally, we confirmed that the lowered expressions of the transcript and protein levels of ABI3 brought on by elevated temperature were correlated with warmth-induced early flowering phenotypes. In conclusion, our data suggest that the BR- and warmth-mediated regulation of ABI3 are important in thermomorphic reproductive phase transitions in plants.

Gene Expression Profiling in Rice Infected with Rice Blast Fungus using SAGE

  • Kim, Sang-Gon;Kim, Sun-Tae;Kim, Sung-Kun;Kang, Kyu-Young
    • The Plant Pathology Journal
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    • v.24 no.4
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    • pp.384-391
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    • 2008
  • Rice blast disease, caused by the pathogenic fungus Magnaporthe grisea, is a serious issue in rice (Oryza sativa L.) growing regions of the world. Transcript profiling in rice inoculated with the fungus has been investigated using the transcriptomics technology, serial analysis of gene expression (SAGE). Short sequence tags containing sufficient information which are ten base-pairs representing the unique transcripts were identified by SAGE technology. We identified a total of 910 tag sequences via the GenBank database, and the resulting genes were shown to be up-regulated in all functional categories under the fungal biotic stress. Compared to the compatible interaction, the stress and defense genes in the incompatible interaction appear to be more up-regulated. Particularly, thaumatin-like gene (TLP) was investigated in determining the gene and protein expression level utilizing Northern and Western blotting analyses, resulting in an increase in both the gene and the protein expression level which arose earlier in the incompatible interaction than in the compatible interaction.

Development and Validation of the Custom Human cDNA Microarray (KISTCHIP-400) for Monitoring Expression of Genes involved in Hormone Disruption

  • Kim, Youn-Jung;Chang, Suk-Tai;Yun, Hye-Jung;Jeon, Hee-Kyung;Ryu, Jae-Chun
    • Proceedings of the Korea Society of Environmental Toocicology Conference
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    • 2003.05a
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    • pp.180-180
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    • 2003
  • Transcript profiling is a particularly valuable tool in the field of steroid receptor biology, as these receptors are ligand-activated transcription factors and therefore exert their initial effects through altering gene expression in responsive cells. Also, an increased awareness of endocrine disrupting chemicals (EBCs) and their potential to affect wildlife and humans has produced a demand for practical screening methods to identify endocrine activity. Here we developed an in-house cDNA microarray, named KISTCHIP-400, with 401 clones, hormone related genes, factors, and ESTs, based on public database and research papers. Theses clones contained estrogen, androgen, thyroid hormone St receptors, sex hormone signal transduction & regulation, c-fos, c-myc, ps2 gene, metabolism related genes etc. And to validate the KISTCHIP-400, we investigated gene expression profiles with reference hormones, 10$\^$-8/ M 17be1a-estradiol, 10$\^$-7/ M testosterone, 10$\^$-7/ M progesterone, and thyroxin in MCF-7 cell line. Although it is in first step of validation, low doses and combinations of EDCs need to be tested. Our preliminary results that indicate the developed microarray may be a useful laboratory tool for screening EDCs and elucidating endocrine disrupting mechanism.

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Characterization of X-linked RNA Transcripts in Matured Bovine Spermatozoa

  • Jeon, Byeong-Gyun;Kumar, B. Mohana;Rho, Gyu-Jin
    • Reproductive and Developmental Biology
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    • v.35 no.3
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    • pp.209-214
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    • 2011
  • Although the function and utility of RNA transcripts derived from matured spermatozoa remains unclear, they might play important roles in the establishment of a paternal genome and subsequently embryo development. Herein, we investigated the expression of X-chromosome linked RNA transcripts in matured bovine spermatozoa. The total RNA was extracted from the matured spermatozoa, and then converted to cDNA. Autosomal genes (ACT-${\beta}$ and H-2A) and X-chromosome linked genes (ANT3, HPRT, MeCP2, RPS4X, XIAP, XIST and ZFX) were analyzed for the characterization of X-chromosome linked RNA transcripts and compared to female fibroblasts by RT-PCR. The transcripts of autosomal genes (ACT-${\beta}$ and H2A) and X-chromosome linked genes (ANT3, HPRT, MeCP2, RPS4X and ZFX) were not detected in spermatozoa. However, XIAP (X-linked inhibitor of apoptosis protein) and XIST (X-chromosome inactive-specific transcript, a kind of paternal imprinted gene) transcripts were detected in spermatozoa, and relative levels of XIAP and XIST transcripts were similar and 0.5-fold lower when compared to female fibroblasts, respectively. Based on the findings, it is summarized that the presence of RNA transcripts of XIAP and XIST in the isolated spermatozoa may imply their role in inhibition of apoptosis and induction of X-chromosome inactivation in embryo development.

A Study on the Iconological Approach of the Korean Traditional Space Design - Focusing on Regional Prototype and Creative Fantasy - (한국 전통공간디자인의 도상해석학적 접근에 관한 연구 - 지역적 원형과 창조적 환상의 개념을 중심으로 -)

  • Park, Kyung-Ae
    • Korean Institute of Interior Design Journal
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    • v.17 no.6
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    • pp.120-127
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    • 2008
  • Korean space design image is a kind of writing as well as one of the sign that dissembles itself as a direct transcript of what it represents. Moreover it is pictorial representation and notions such as mental and perceptual imaginary. Significance of Iconology lies in how we transform image and the imagination that produces it into power of trust and respect. From this point of view, the process of this study is illustrated as follows: At first, this study search out concept of archetype, collective unconsciousness and collective representation that found principles on basic theory for interpretation of korean space icon. Secondly, it mentions theoretical background of iconological contents and structure. And it clarifies Iconology as a method that is applicable logic for Korean space design. Finally, as an analysis of korean space design, this study analyse in three steps that are pre-iconological description, iconological analysis, iconological interpretation each in terms of modernization at regional korean space design. In the step of the pre-iconological description, it describe visual representative style based on era and place. In the step of the iconological analysis, the typical structure is classified in status, vernacular, ethnic, traditional. In the step of the iconological interpretation, connotation is categorized into allegory, multivalence, potential. Through this process, this study suggest that iconology is an appropriate analysis system of Korean space design images that represent symbols combined with our collective emotion.