• Title/Summary/Keyword: sequence length

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Phylogenetic Study of Korean Chrysosplenium Based on nrDNA ITS Sequences (ITS 염기서열에 의한 한국산 괭이눈속(Chrysosplenium)의 계통학적 연구)

  • Han, Jong-Won;Yang, Sun-Gyu;Kim, Hyun-Jun;Jang, Chang-Gee;Park, Jeong-Mi;Kang, Shin-Ho
    • Korean Journal of Plant Resources
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    • v.24 no.4
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    • pp.358-369
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    • 2011
  • The internal transcribed spacer (ITS) regions of nuclear ribosomal DNA from genus Chrysosplenium were sequenced to address phylogenetic relationship. ITS including 5.8S sequence varied in length from 647 bp to 653 bp. Among them, 219 sites were variable sites with parsimony-informative. The aligned sequences were analyzed by maximum parsimony (MP) and neighbor-joining (NJ) methods. In the strict consensus trees of parsimony analysis, the monophyly of Chrysosplenium was supported by 100% bootstrap value. The first clade, C. pseudofauriei was at the basal position of the genus, and others formed two clades with high bootstrap support. The second clade included Ser. Pilosa and Ser. Oppositifolia and third clade included Ser. Alternifolia and Ser. Flagellifera. The NJ trees showed essentially the same topology. Finally, DNA sequences of ITS regions were useful phylogenetic marker in this genus. Based on the ITS and ridge seed morphological results, C. sphaerospermum Maxim. and C. valdepilosum (Ohwi) S.H. Kang & J.W. Han were discussed their scientific names and taxonomic positions.

Pharmaco-medical Application of Antimicrobial Peptides Derived from Insect (곤충유래 항균 펩티드의 의약학적 적용)

  • Lee, Joon Ha;Kim, In-Woo;Kim, Mi-Ae;Yun, Eun Young;Hwang, Jae Sam
    • Journal of Life Science
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    • v.26 no.6
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    • pp.737-748
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    • 2016
  • By this time, insect antimicrobial peptides (AMPs) have been characterized more than 150 peptides since purification of cecropin in the hemolymph of pupae from Hyalophora cecropia in 1980. Therefore, it is considered that insects are good sources of AMP selection. Insect AMPs are small (low molecular weight) and cationic, and amphipathic with variable length, sequence, and structure. They perform a pivotal role on humoral immunity in the insect innate immune system against invading pathogens such as bacteria, fungi, parasites, and viruses. Most of the insect AMPs are induced rapidly in the fat bodies and other specific tissues of insects after septic injury or immune challenge. Then the AMPs subsequently released into the hemolymph to act against microorganisms. These peptides have a broad antimicrobial spectrum against various microbes including anticancer activities. Insect AMPs could be divided into four families based on their structures and sequences. That is the α-helical peptides, cysteine-rich peptides, proline-rich peptides, and glycine-rich peptides/proteins. For instance, cecropins, insect defensins, proline-rich peptides, and attacins are common insect AMPs, but gloverins and moricins have been identified only in lepidopteran species. This review focuses on AMPs from insects and discusses current knowledge and recent progress with potential applications of insect AMPs.

Molecular Cloning and Expression of the Metallothionein Gene under Environmental Stresses in Sweet Potato (고구마 metallothionein 유전자의 클로닝 및 환경 스트레스 하에서 발현 분석)

  • Kim, Young-Hwa;Yu, Eun Jeong;Huh, Gyung-Hye
    • Journal of Life Science
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    • v.27 no.12
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    • pp.1415-1420
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    • 2017
  • The metallothionein (MT) gene (IbMT3) was selected from an EST library of suspension-cultured sweet potato cells. The MT gene, which is one of abundant ESTs in the library, is involved in stress regulation of cells and tissues. A full-length IbMT3 cDNA was obtained and analysis of its nucleotide sequence revealed that IbMT3 encoded a type 3 MT protein, based on its structural characteristics. The function of type 3 MT in plants is not yet known. Northern blot analysis showed stronger expression of IbMT3 in suspension-cultured cells than in sweet potato plant leaves. Since cell culture is known to impose a state of oxidative stress on cells, sweet potato plants were subjected to oxidative stress to investigate the transcriptional regulation of IbMT3. When the herbicide methyl viologen (MV) was administered for 6, 12, and 24 hr, IbMT3 transcription rapidly increased at 6 hr and then decreased. A cold treatment at $15^{\circ}C$ for 24 and 48 hr resulted in a gradual increase in IbMT3 expression. These findings indicate that IbMT3 expression is regulated in response to environmental and oxidative stress. IbMT3 isoform is expected to have antioxidant effects in sweet potato plants and may play an important role in cellular adaptation to oxidative stress.

Phylogeny of Ganoderma Based on the Restriction Enzyme Analysis of Mitochondrial DNA (미토콘드리아 DNA의 제한효소 분석법에 의한 영지의 계통분류)

  • Hong, Soon-Gyu;Jung, Hack-Sung
    • Korean Journal of Microbiology
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    • v.32 no.4
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    • pp.245-251
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    • 1994
  • Ten strains of 7 species from the genus Ganoderma, G. lucidum ATCC 64251, FP-103561-T, and ES70701, G. applanatum ATCC 44053 and FP-57035-T. G. lobatum ATCC 42985, G. resinaceum ATCC 52416, G. subamboinense var. laevisporum ATCC 52420, G. meredithae ATCC 64492, and G. microsporum ATCC 76024, were studied to discuss their phylogenetic relationships by utilizing restriction fragment length polymorphisms (RFLPs) of mitochondrial DNAs (mtDNAs). Six restriction enzymes, BamHI, BglII, EcoRI, HindIII, PvuII, and XbaI which digested mtDNAs into adequate numbers of restriction fragments for cluster analysis, were used in this study. Restriction profiles of strains for each restriction enzyme were treated as analysis characters to calculate similarity coefficients, which were converted into nucleotide sequence divergence values whose mean values were then arranged in a matrix table. This table was utilized for a phylogenetic analysis using the Neighborjoining method of the PHYLIP package to construct phylogenetic tree. Three strains of G. lucidum and two strains of G. applanatum exhibited different lineages each but one of G. applanatum strains showed a close relationship with G. lobatum, which reflected the species complexity of these species whose strains were phenotypically indistinguishable but genetically distinct. The present results suggest that the natural classification of Ganoderma needs to be considered from the viewpoints of molecular biology-based systematics as well as morphological classifications and cultural identifications for better phylogenetic conclusions.

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Molecular Cloning of Mutant cDNA of PU.1 Gene (PU.1 유전자(cDNA)의 인위적 변이체 클로닝)

  • 류종석;유시현
    • KSBB Journal
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    • v.10 no.5
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    • pp.499-509
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    • 1995
  • PU.1, a tissue-specific transcription activator, binds to a purine-rich sequence(5'-GAGGAA-3') called PU box. The PU.1 cDNA consists of an open reading frame of 816 nucleotides coding for 272 amino acids. The amino terminal end is highly acidic, while the carboxyl terminal end is highly basic. Transcriptional activation domain is located at the amino terminal end, while DNA binding domain is located at the carboxyl terminal end. Activation of PU.1 transcription factor is supposed to be accomplished by the phosphorylation of serine residue(s). There exist 22 serines in the PU.1. Five(the 41, 45, 132$.$133, and 148th) of the serines(plausible phosphorylation site by casein kinase II), are the primary targets of interest in elucidating the molecular mechanism(s) of the action of the PU.1 gene. In this study, PU.1 cDNA coding for the five serine residues(41th AGC, 45th AGC, 132$.$133th AGC$.$TCA, and 148th TCT), was mutated to alanine codon(41th GCC, 45th GCC, 132$.$133th GCC$.$GCA, and 1481h GCT), respectively, by Splicing-Overlapping-Extension(SOE) using Polymerase Chain Reaction(PCR). And each mutated cDNA fragments was ligated into pBluescript KS+ digested with HindIII and Xba I, to generate mutant clones named pKKS41A, pRKS45A, pMKS132$.$133A, and pMKS148A. The clones will be informative to study the "Structure and Function" of the immu-nologically important gene, PU.1.

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Trajectory Clustering in Road Network Environment (도로 네트워크 환경을 위한 궤적 클러스터링)

  • Bak, Ji-Haeng;Won, Jung-Im;Kim, Sang-Wook
    • The KIPS Transactions:PartD
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    • v.16D no.3
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    • pp.317-326
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    • 2009
  • Recently, there have been many research efforts proposed on trajectory information. Most of them mainly focus their attention on those objects moving in Euclidean space. Many real-world applications such as telematics, however, deal with objects that move only over road networks, which are highly restricted for movement. Thus, the existing methods targeting Euclidean space cannot be directly applied to the road network space. This paper proposes a new clustering scheme for a large volume of trajectory information of objects moving over road networks. To the end, we first define a trajectory on a road network as a sequence of road segments a moving object has passed by. Next, we propose a similarity measurement scheme that judges the degree of similarity by considering the total length of matched road segments. Based on such similarity measurement, we propose a new clustering algorithm for trajectories by modifying and adjusting the FastMap and hierarchical clustering schemes. To evaluate the performance of the proposed clustering scheme, we also develop a trajectory generator considering the observation that most objects tend to move from the starting point to the destination point along their shortest path, and perform a variety of experiments using the trajectories thus generated. The performance result shows that our scheme has the accuracy of over 95% in comparison with that judged by human beings.

Alkaliphilic Endoxylanase from Lignocellulolytic Microbial Consortium Metagenome for Biobleaching of Eucalyptus Pulp

  • Weerachavangkul, Chawannapak;Laothanachareon, Thanaporn;Boonyapakron, Katewadee;Wongwilaiwalin, Sarunyou;Nimchua, Thidarat;Eurwilaichitr, Lily;Pootanakit, Kusol;Igarashi, Yasuo;Champreda, Verawat
    • Journal of Microbiology and Biotechnology
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    • v.22 no.12
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    • pp.1636-1643
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    • 2012
  • Enzymatic pre-bleaching by modification of pulp fibers with xylanases is an attractive approach to reduce the consumption of toxic bleaching chemicals in the paper industry. In this study, an alkaliphilic endoxylanase gene was isolated from metagenomic DNA of a structurally stable thermophilic lignocellulose-degrading microbial consortium using amplification with conserved glycosyl hydrolase family 10 primers and subsequent genome walking. The full-length xylanase showed 78% sequence identity to an endo-${\beta}$-1,4-xylanase of Clostridium phytofermentans and was expressed in a mature form with an N-terminal His6 tag fusion in Escherichia coli. The recombinant xylanase Xyn3F was thermotolerant and alkaliphilic, working optimally at $65-70^{\circ}C$ with an optimal pH at 9-10 and retaining >80% activity at pH 9, $60^{\circ}C$ for 1 h. Xyn3F showed a $V_{max}$ of 2,327 IU/mg and $K_m$ of 3.5 mg/ml on birchwood xylan. Pre-bleaching of industrial eucalyptus pulp with no prior pH adjustment (pH 9) using Xyn3F at 50 IU/g dried pulp led to 4.5-5.1% increase in final pulp brightness and 90.4-102.4% increase in whiteness after a single-step hypochlorite bleaching over the untreated pulp, which allowed at least 20% decrease in hypochlorite consumption to achieve the same final bleaching indices. The alkaliphilic xylanase is promising for application in an environmentally friendly bleaching step of kraft and soda pulps with no requirement for pH adjustment, leading to improved economic feasibility of the process.

Identification of Arbuscular Mycorrhizal Fungi Colonizing Panax ginseng Using 18S rDNA Sequence (18S rDNA를 이용한 인삼(Panax ginseng)의 내생균근 균의 동정)

  • Eo, Ju-Kyeong;Kim, Dong-Hun;Jeong, Hyeon-Suk;Eom, Ahn-Heum
    • Applied Biological Chemistry
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    • v.47 no.2
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    • pp.182-186
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    • 2004
  • Morphological observation of roots and molecular technique were used to investigate the symbiotic relationships between arbuscular mycorrhizal (AM) fungi and ginseng roots. Korean ginseng, Panax ginseng, was collected from 8 sites in Korea. Colonization pattern of AM fungi in ginseng roots was determined as an Arum type under light microscopes. Nested PCR using AM fungal specific primers was employed to amplify a partial region on 18s rDNA of AM fungi from the root extracted mixed DNA. The amplified DNA was cloned and analyzed by random fragment length polymorphism (RFLP) with restriction enzymes, AluI, HinfI and AsuC21. One from each RFLP pattern was selected for sequencing. A total 16 clones were sequenced and identified as 2 species of AM fungi; Paraglomus brasilianum and Glomus spurcum. Paramglomus brasilianum was found from most of the ginseng roots, in this syudy suggesting that this species of AM fungi could have specific relationship with the ginseng root. Possible roles of AM fungal species in ginseng roots are discussed.

The Wet and Dry Etching Process of Thin Film Transistor (박막트랜지스터의 습식 및 건식 식각 공정)

  • Park, Choon-Sik;Hur, Chang-Wu
    • Journal of the Korea Institute of Information and Communication Engineering
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    • v.13 no.7
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    • pp.1393-1398
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    • 2009
  • Conventionally, etching is first considered for microelectronics fabrication process and is specially important in process of a-Si:H thin film transistor for LCD. In this paper, we stabilize properties of device by development of wet and dry etching process. The a-Si:H TFTs of this paper is inverted staggered type. The gate electrode is lower part. The gate electrode is formed by patterning with length of 8 ${\mu}$m${\sim}$16 ${\mu}$m and width of 80${\sim}$200 ${\mu}$m after depositing with gate electrode (Cr) 1500 ${\AA}$under coming 7059 glass substrate. We have fabricated a-SiN:H, conductor, etch-stopper and photo resistor on gate electrode in sequence, respectively. The thickness of these thin films is formed with a-SiN:H (2000 ${\mu}$m), a-Si:H(2000 ${\mu}$m) and n+a-Si:H (500 ${\mu}$m), We have deposited n-a-Si:H, NPR(Negative Photo Resister) layer after forming pattern of Cr gate electrode by etch-stopper pattern. The NPR layer by inverting pattern of upper gate electrode is patterned and the n+a-Si:H layer is etched by the NPR pattern. The NPR layer is removed. After Cr layer is deposited and patterned, the source-drain electrode is formed. In the fabricated TFT, the most frequent problems are over and under etching in etching process. We were able to improve properties of device by strict criterion on wet, dry etching and cleaning process.

Molecular Cloning and Expression of a Cu/Zn-Containing Superoxide Dismutase from Thellungiella halophila

  • Xu, Xiaojing;Zhou, Yijun;Wei, Shanjun;Ren, Dongtao;Yang, Min;Bu, Huahu;Kang, Mingming;Wang, Junli;Feng, Jinchao
    • Molecules and Cells
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    • v.27 no.4
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    • pp.423-428
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    • 2009
  • Superoxide dismutases (SODs) constitute the first line of cellular defense against oxidative stress in plants. SODs generally occur in three different forms with Cu/Zn, Fe, or Mn as prosthetic metals. We cloned the full-length cDNA of the Thellungiella halophila Cu/Zn-SOD gene ThCSD using degenerate RT-PCR and rapid amplification of cDNA ends (RACE). Sequence analysis indicated that the ThCSD gene (GenBank accession number EF405867) had an open reading frame of 456 bp. The deduced 152-amino acid polypeptide had a predicted molecular weight of 15.1 kDa, an estimated pI of 5.4, and a putative Cu/Zn-binding site. Recombinant ThCSD protein was expressed in Escherichia coli and assayed for SOD enzymatic activity in a native polyacrylamide gel. The SOD activity of ThCSD was inactivated by potassium cyanide and hydrogen peroxide but not by sodium azide, confirming that ThCSD is a Cu/Zn-SOD. Northern blotting demonstrated that ThCSD is expressed in roots, stems, and leaves. ThCSD mRNA levels increased by about 30-fold when plants were treated with sodium chloride (NaCl), abscisic acid (ABA), and indole-acetic acid (IAA) and by about 50-fold when treated with UVB light. These results indicate that ThCSD is involved in physiological pathways activated by a variety of environmental conditions.