• Title/Summary/Keyword: base-isolated

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Isolation and Characterization of a Type II Peroxiredoxin Gene from Panax ginseng C. A. Meyer

  • Kim, Yu-Jin;Lee, Jung-Hye;Lee, Ok-Ran;Shim, Ju-Sun;Jung, Seok-Kyu;Son, Na-Ri;Kim, Ju-Han;Kim, Se-Young;Yang, Deok-Chun
    • Journal of Ginseng Research
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    • v.34 no.4
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    • pp.296-303
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    • 2010
  • A peroxiredoxin cDNA (PgPrx) was isolated and characterized from the leaves of Panax ginseng. The cDNA is 716 nucleotides long and has an open reading frame of 489 base pairs with a deduced amino acid sequence of 162 residues. The calculated molecular mass of the mature protein is approximately 17.4 kDa with a predicted isoelectric point of 5.37. A GenBank BlastX search revealed that the deduced amino acid sequence of PgPrx shares a high degree homology with type II peroxiredoxin (Prx) proteins in other plants. The PgPrx gene was highly expressed in leaves, and expressed at a low level in the stem. To analyze the gene expression of PgPrx in response to various abiotic stresses, we utilized real-time quantitative RT-PCR. Our results reveal that PgPrx expression is induced by ultraviolet irradiation, low temperature, and salt. The induction of PgPrx in response to abiotic stimuli suggests that ginseng Prx may function to protect the host against environmental stresses.

Ancestral Remnants in the Deoxyribonucleic Acid from Pseudomonas (Deoxyribonucleic Acid 속의 Pseudomonas로부터의 Ancestral Remnants)

  • J. De Ley;In Won Park
    • Journal of the Korean Chemical Society
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    • v.13 no.4
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    • pp.355-364
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    • 1969
  • Cross-hybridizations between DNA of two pseudomonads and a xanthomonad suggested that the three DNA types had a considerable section in common. The existence of this common part was proved by hybridization of preselected DNA, i.e. DNA resulting from a previous hybridization between any one set of two DNA types, with the third type. It was thus shown that about 50% of the DNA of the three organisms was similar. This common part was isolated in pure state and its % (G+C) was found to be indentical to the overall base composition of the native DNA. The evolutionary drift in % (G+C) could thus not be detected. The total molecular weight of the chromosornal DNA/bacterial nucleoid was determined to be 2.4 ${\times} 10^9$daltons. It can therefore be estimated that the common putida-fluorescenspelargonii DNA part consists of some 2,000 cistrons. P. putida and P. fluorescens share an additional 1,300 cistrons, and all xanthomonads share at least an additional 1,000 cistrons.

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The transposition pattern of the Ac element and its use for targeted transposition in Arabidopsis thaliana

  • Machida, Yasunori;Onouchi, Hitoshi;Tanaka, Hirokazu;Hamada, Susumu;Ishikawa, Takaaki;Semiarti, Endang;Iwakawa, Hidekazu;Nomura, Kiyohito;Machida, Chiyoko
    • Proceedings of the Botanical Society of Korea Conference
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    • 1999.07a
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    • pp.11-15
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    • 1999
  • In order to evlauate feasibility of the gene tagging by the maize transposable element Ac in heterologous plant systems, we have investigated physical distances and directions of transposition of the element in Arabidopsis thaliana and tobacco cultured cell line BY-2. We prepared a T-DNA construct that carried a non-autonomous derivative of Ac with a site for cleavage by endonuclease I-Scel (designated dAc-I-RS element). Another cleavage site was also introduced into the T-DNA region outside dAc-I-RS. A number of transgenic Arabidopsis plants were generated, each of which had a single copy of the T-DNA at a different chromosomal location. To examine the pattern of transposition, three out of these transgenic plants were crossed with the Arabidopsis plant that carried the gene for Ac transposase and progeny in which dAc-I-RS had been transposed were isolated. After digestion of the genomic DNA of these progeny with I-SceI, sizes of segment of DNA were determined byd pulse-field gel electrophoresis. We also performed linkage analysis for the transposed elements and sites of mutations near the elements. Our results with three transgenic lines showed that 50% of all transposition events had occurred within 1,700 kilo-base pairs (kb) on the same chromosome, with 35% within 200 kb, and that the elements transposed in both directions on the chromosome with roughly equal probability. The data thus indicate that the Ac-Ds system is most useful for tagging of genes that are present within 200 kb of the chromosomal site of Ac in Arabidopsis. In addition, determination of the precise localization of the transposed dAc-I-RS element should definitely assist in map-based cloning of genes around insertion sites. In the present paper, we report typical examples of such gene isolation studies.

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세포내의 물의 상태

  • 강사욱
    • Proceedings of the Botanical Society of Korea Conference
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    • 1985.08b
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    • pp.51-57
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    • 1985
  • In order to evlauate feasibility of the gene tagging by the maize transposable element Ac in heterologous plant systems, we have investigated physical distances and directions of transposition of the element in Arabidopsis thaliana and tobacco cultured cell line BY-2. We prepared a T-DNA construct that carried a non-autonomous derivative of Ac with a site for cleavage by endonuclease I-Scel (designated dAc-I-RS element). Another cleavage site was also introduced into the T-DNA region outside dAc-I-RS. A number of transgenic Arabidopsis plants were generated, each of which had a single copy of the T-DNA at a different chromosomal location. To examine the pattern of transposition, three out of these transgenic plants were crossed with the Arabidopsis plant that carried the gene for Ac transposase and progeny in which dAc-I-RS had been transposed were isolated. After digestion of the genomic DNA of these progeny with I-SceI, sizes of segment of DNA were determined byd pulse-field gel electrophoresis. We also performed linkage analysis for the transposed elements and sites of mutations near the elements. Our results with three transgenic lines showed that 50% of all transposition events had occurred within 1, 700 kilo-base pairs (kb) on the same chromosome, with 35% within 200 kb, and that the elements transposed in both directions on the chromosome with roughly equal probability. The data thus indicate that the Ac-Ds system is most useful for tagging of genes that are present within 200 kb of the chromosomal site of Ac in Arabidopsis. In addition, determination of the precise localization of the transposed dAc-I-RS element should definitely assist in map-based cloning of genes around insertion sites. In the present paper, we report typical examples of such gene isolation studies.

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Sequence analysis of the variable VP2 gene of infectious bursal disease viruses isolated in Korea (Infectious bursal disease virus(국내분리주)의 variable VP2 gene의 분석)

  • Kwon, Hyuk-moo;Kim, Dae-kyu;Seong, Hwan-woo
    • Korean Journal of Veterinary Research
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    • v.39 no.3
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    • pp.545-553
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    • 1999
  • A 474-base pair segment covering the hypervariable region of VP2 gene from six Korean infectious bursal disease virus(K-IBDV) isolates(K1, K2, SH/92, 225, 269, 310) and one attenuated IBDV(DAE) were amplified using RT-PCR, sequenced, and compared with published sequences for IBDV. K-IBDV isolates(K1, K2, SH/92, 225, 269) and foreign very virulent(vv) IBDV strains had 94.93~100% amino acid sequence similarity. K-IBDV isolate 310 and other K-IBDV isolates had 84.31~86.07% amino acid sequence similarity. Attenuated strain(DAE), like other attenuated strain, has substitution at positions 279(D to N) and 284(A to T) as well as in the serine-rich heptapeptide region. Five K-IBDV isolates except 310 isolate share unique amino acid residues at positions 222(A), 256(I), 294(I) which are not present in other standard and attenuated strains. At the two hydrophilic region, K-IBDV isolates except 310 isolate had identical amino acids comparing with Belgium vv IBDV 894VB but had four amino acid substitutions comparing with Chinese vv IBDV F9502. The SWSASGS heptapeptide is conserved in all K-IBDV isolates. The sequence of K-IBDV isolate 310 was markedly different from other IBDV strains, evolving from a separate lineage than the others. By phylogenetic analysis, Five K-IBDV isolates except 310 isolate were categorized in one group with foreign vv IBDV isolates but K-IBDV isolate 310 was categorized in a separate group which was differentiated from other compared IBDV strains.

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Molecular Cloning of a Gene Encoding $\alpha$-L-Arabinofuranosidase from Hyperthermophile Thermotoga maritima and Characterization of Its Biochemical Properties

  • Keum, In-Kyung;Lee, Eun-Joo;Kim, Tae-Jip;Kim, Chung-Ho;Han, Nam-Soo
    • Proceedings of the Korean Society for Applied Microbiology Conference
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    • 2004.06a
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    • pp.273-277
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    • 2004
  • $\alpha$-L-Arabinofuranosidase ($\alpha$-L-AFase, EC 3.2.1.55) was isolated from hyperthermophilic microorganism, Thermotoga maritima. The open reading frame (ORF) of $\alpha$-L-AFase gene is 1,455 bp long and encodes 484 amino acid residues with a molecular weight of 55,265 Da. The ORF of $\alpha$-L-AFase gene was introduced into the E. coli expression vector, $_p/RSET-B, and overexpressed in E. coli BL21. The purified recombinant $\alpha$-L-AFase showed the highest activity at 10$0^{\circ}C$ and pH 5.5. The purified enzyme appeared to have no metal cofactor requirement. The Km and specific activity values of the recombinant enzyme were 0.99 mM and 1,200 U/mg on p-nitrophenyl-$\alpha$-L-arabinofuranoside. It released only L-arabinose from sugar beet arabinan, sugar beet debranched arabinan and oat spelts arabinoxylan but had no activity onarabinogalactan and gum arabic. This result suggests that L-arabinose could be produced from natural polysaccharides using this enzyme. Mutant enzymes which Glu26, Glu172 and Glu281 residues were replaced to alanine, aspartic acid or glutamine caused Kcat to decrease by a factor of between 10$^3$ and 10$^4$. Glu172 and Glu281 residues of $\alpha$-L-AFase are seemed to be the acid/base and nucleophile in catalytic reaction, respectively, and Glu26 is supposed to playa key role in substrate binding.ng.

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Approach case design for achieve the social values (사회적 가치 실현을 위한 디자인의 접근 사례)

  • Kim, Myoung Yun
    • Smart Media Journal
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    • v.3 no.1
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    • pp.46-51
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    • 2014
  • The focus of design in the contemporary point of view has changed from the improvement in production efficiency for profitability and in styling for sales promotion to human-oriented design. Human-centered design has developed into design which includes social and ethical responsibility and values. With tradition and culture as a base the principles of markets and capital are incorporated as well. Such current design is the outcome of design which honors social values represented based on culture acquired from individuals and society and the the responsibility of the designer who provides it. For the realization of social values in design, it should be considered along with the influence of design on society, this study presented the following cases: First, human-oriented design for those who are isolated, but should not be ignored; Second, cultural-oriented design which makes a contribution to local conditions; and third, usability-oriented design which replaces familiar ideas with new paradigms. This study presented the function and meaning of design in terms of purpose and the methods to solve social and human problems using design. Further studies should deal with the realization and extension of social values through design.

An investigation of Panax ginseng Meyer growth promotion and the biocontrol potential of antagonistic bacteria against ginseng black spot

  • Sun, Zhuo;Yang, Limin;Zhang, Lianxue;Han, Mei
    • Journal of Ginseng Research
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    • v.42 no.3
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    • pp.304-311
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    • 2018
  • Background: Ginseng black spot disease resulting from Alternaria panax Whuetz is a common soil-borne disease, with an annual incidence rate higher than 20-30%. In this study, the bacterial strains with good antagonistic effect against A. panax are screened. Methods: A total of 285 bacterial strains isolated from ginseng rhizosphere soils were screened using the Kirby-Bauer disk diffusion method and the Oxford cup plate assay. We analyzed the antifungal spectrum of SZ-22 by confronting incubation. To evaluate the efficacy of biocontrol against ginseng black spot and for growth promotion by SZ-22, we performed pot experiments in a plastic greenhouse. Taxonomic position of SZ-22 was identified using morphology, physiological, and biochemical characteristics, 16S ribosomal DNA, and gyrB sequences. Results: SZ-22 (which was identified as Brevundimonas terrae) showed the strongest inhibition rate against A. panax, which showed 83.70% inhibition, and it also provided broad-spectrum antifungal effects. The inhibition efficacies of the SZ-22 bacterial suspension against ginseng black spot reached 82.47% inhibition, which is significantly higher than that of the 25% suspension concentrate azoxystrobin fungicide treatment (p < 0.05). Moreover, the SZ-22 bacterial suspension also caused ginseng plant growth promotion as well as root enhancement. Conclusion: Although the results of the outdoor pot-culture method were influenced by the pathogen inoculum density, the cropping history of the field site, and the weather conditions, B. terrae SZ-22 controlled ginseng black spot and promoted ginseng growth successfully. This study provides resource for the biocontrol of ginseng black spot.

Protein Carboxylmethylation in Porcine Spleen is Mainly Mediated by Class I Protein Carboxyl O-Methyltransferase

  • Cho, Jae-Youl;Kim, Sung-Soo;Kwon, Myung-Hee;Kim, Seong-Hwan;Lee, Hyang-Woo;Hong, Sung-Youl
    • Archives of Pharmacal Research
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    • v.27 no.2
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    • pp.206-216
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    • 2004
  • The functional role of protein carboxylmethylation (PCM) has not yet been clearly elucidated in the tissue level. The biochemical feature of PCM in porcine spleen was therefore studied by investigating the methyl accepting capacity (MAC) of natural endogenous substrate proteins for protein carboxyl O-methyltransferase (PCMT) in various conditions. Strong acidic and alkaline-conditioned (at pH 11.0) analyses of the MAC indicated that approximately 65% of total protein methylation seemed to be mediated by spleen PCMT. The hydrolytic kinetics of the PCM products, such as carboxylmethylesters (CMEs), under mild alkaline conditions revealed that there may be three different kinds of CMEs [displaying half-times (T$_{1}$2/) of 1.1 min (82.7% of total CMEs), 13.9 min (4.6%), and 478.0 min (12.7%)], assuming that the majority of CME is base-labile and may be catalyzed by class I PCMT. In agreement with these results, several natural endogenous substrate proteins (14, 31 and 86 kDa) were identified strikingly by acidic-conditioned electrophoresis, and their MAC was lost upon alkaline conditions. On the other hand, other proteins (23 and 62 kDa) weakly appeared under alkaline conditions, indicating that PCM mediated by class II or III PCMT may be a minor reaction. The MAC of an isolated endogenous substrate protein (23-kDa) was also detected upon acidic-conditioned electrophoresis. Therefore, our date suggest that most spleen PCM may be catalyzed by class I PCMT, which participates in repairing aged proteins.

Studies on Aromatic Dihydrazines (I). Synthesis of p-Phenylenedihydrazine via Tetrazonium Salt and Formation of Dihydrazones (芳香族 디히드라진에 關한 硏究 (第 1 報). 테트라아조늄염을 通한 파라페닐렌디히드라진의 합성 및 디히드라존의 生成)

  • Woo Young Lee
    • Journal of the Korean Chemical Society
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    • v.18 no.1
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    • pp.50-57
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    • 1974
  • A practical method applicable to the synthesis of aromatic dihydrazines was proposed by reducing tetrazonium salt in strong mineral acid media. By diazotizing p-phenylenediamine with sodium nitrite in a medium of concentrated hydrochloric acid or 45 % perchloric acid at $-5 {\sim} -10{\circ}C$ and reducing the tetrazonium salt with stannous chloride, p-phenylenedihydrazine (PPDH) was separated in the form of hydrochloride as colorless fine needles. Since PPDH was subject to oxidation and unstable, the free base could not be isolated. PPDH${\cdot}$2HCl was decomposed at $^180{\circ}C$ without showing sharp melting point. It behaved largely as aromatic monohydrazines, and reacted immediately with aldehydes and ketones in acetate buffer, giving generally yellow to brownish condensation products, dihydrazones. This suggests that PPDH will react with dicarbonyl compounds producing high molecular polymers or cyclization products.

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