• Title/Summary/Keyword: ITS sequence

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Levan-Producing Bacillus subtilis BS 62 and Its Phylogeny Based on Its 16S rDNA Sequence

  • Choi, Seong-Hyun;Chang, Sung;Choi, Woo-Young
    • Journal of Microbiology and Biotechnology
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    • v.11 no.3
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    • pp.428-434
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    • 2001
  • A viscous substance producer strain BS62, which was isolated from conventional Chungkookjang, was examined for its productivity of levansucrase and levan during soybean fermentation at $37{\circ}C$. After one day of cultivation, the enzyme activity reached the highest level, 8 units $ml^{-1}$. Extracts of fermented soybeans were precipitated by ethanol and hydrolyzed by either 0.1 N HCl or invertase, and the hydrolyzates were analyzed using thin layer and ion chromatographies. Fructose was the only sugar detected. This suggest that fructose was derived from the levan produced by the strain BS62 during soybean fermentation. The aerobic, endospore-forming bacterium BS62 was identified as a Bacillus subtilis sp., based on the composition of its cellular fatty acids and phylogeny, which was determined by its 16S rDNA sequence.

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Characterization of a Brown Rot Fungus Isolated from Dwarf Flowering Almond in Korea

  • Shim, Myoung-Yong;Jeon, Young-Jae;Kim, Seong-Hwan
    • Mycobiology
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    • v.35 no.1
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    • pp.30-35
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    • 2007
  • The fruits showing brown rot symptom on dwarf flowering almond were found in Gongju, Chungchungnam-Do in Korea in July 2005. Small water-soaked lesions on the fruits were initiated, and gradually developed to soft rot covered with gray conidia. Then the diseased fruits were shrunk and became grayish-black mummies. A fungus was isolated from the diseased fruit and its morphological, cultural and molecular genetic characteristics were investigated. Typical blastospores of Monilinia spp. were observed under a light microscope both from tissues of the diseased fruits and from PDA-grown cultures. The fungus grew well at $25^{\circ}C$ and on PDA. The ITS ribosomal DNA region (650 bp) of the fungus was amplified by PCR and analyzed. Comparative data on ITS sequence homology among Monilinia spp., ITS sequence-based phylogram and morphological characteristics showed that the fungus is Monilinia fructicola. This is the first report on Monilinia fructicola causing brown rot on fruits of dwarf flowering almond in Korea.

Mutation of a Transposed Amino Acid Triplet Repeat Enhances Coupling of m1 Muscarinic Receptor to Activation of Phospholipase C

  • Lee, Seok-Yong;Cho, Tai-Soon
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1996.04a
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    • pp.206-206
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    • 1996
  • The C-terminus ends of the second putative transmembrane domains of both m1 and m2 muscarinic receptors contain a triplet of amino acid residues consisting of leucine (L), tyrosine (Y) and threonine (T), This triplet is repeated as LYT-LYT in m2 receptors at the interface between the second transmembrane domain and the first extracellular loop. Interestingly, however, it is repeated in a transposed fashion (LYT-TYL) in the sequence of m1 receptors. In this work we employed site-directed mutagenesis to investigate the possible significance of this unique sequence diversity for determining the distinct differential drug-receptor interaction and cellular function at m1 muscarinic receptor. Mutation of the LYTTYL sequence of m1 receptors to the corresponding m2 receptor LYTLYT sequence, however, did not result in a significant change in the binding affinity of the agonist carbachol or in the affinity of the majority of a series of receptor antagonists which are able to discriminate between wild-type m1 and m2 receptors. Surprisingly, the LYTLYT ml receptor mutant demonstrated markedly enhanced coupling to activation of phospholipase C without a change in its coupling to increased cyclic AMP formation. There was also an enhanced receptor sensitivity in transducing elevation of intracellular Ca$\^$2+/. These changes were not due to alterations in the rate of receptor. desensitization or sequestration, On the other hand, the reverse LYTLYT-LYTTYL mutation in the m2 receptor did not alter its coupling to inhibition of adenylate cyclase, but slightly enhanced its coupling to stimulation of PI hydrolysis, Our data suggest that the LYTTYL/LYTLYT sequence difference between ml and n12 muscarinic receptors is not involved in determining receptor pharmacology. On the other hand, while these differences might play a role in the modulation of muscarinic receptor coupling to PI hydrolysis, they are not important for specifying coupling of various subtypes of muscarinic receptors to different cellular signaling pathways.

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A Multiple Sequence Alignment Algorithm using Clustering Divergence (콜러스터링 분기를 이용한 다중 서열 정렬 알고리즘)

  • Lee Byung-ll;Lee Jong-Yun;Jung Soon-Key
    • Journal of the Korea Society of Computer and Information
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    • v.10 no.5 s.37
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    • pp.1-10
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    • 2005
  • Multiple sequence alignment(MSA) is a fundamental technique of DNA and Protein sequence analysis. Biological sequences are aligned vertically in order to show the similarities and differences among them. In this Paper, we Propose an effcient group alignment method, which is based on clustering divergency, to Perform the alignment between two groups of sequences. The Proposed algorithm is a clustering divergence(CDMS)-based multiple sequence alignment and a top-down approach. The algorithm builds the tree topology for merging. It is so based on the concept that two sequences having the longest distance should be spilt into two clusters. We expect that our sequence alignment algorithm improves its qualify and speeds up better than traditional algorithm Clustal-W.

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Partial Sequence Analysis of Puumala Virus M Segment from Bats in Korea

  • Yun, Bo-Kyoung;Yoon, Jeong-Joong;Lee, Yun-Tai
    • The Journal of Korean Society of Virology
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    • v.29 no.1
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    • pp.23-31
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    • 1999
  • Hantavirus is a genus of the Bunyaviridae family causing two serious diseases, hemorrhagic fever with renal syndrome (HFRS) and hantavirus pulmonary syndrome (HPS). Puumala virus is a member of hantavirus originally found in Europe, and its natural reservoir is Clethrionomys glareolus. It is also associated with the human disease nephropathia epidemica, a milder form of HFRS. To identify the hantaviruses in bats, bats were collected from Jeong-Sun, Won-Joo, Chung-Ju and Hwa-Cheon area in Korea, and nested RT-PCR was performed with serotype specific primer from M segment. Interestingly, Puumala virus was detected in bats (Rhinolophus ferrum-equinum) only from Won-Joo. The 327 bp nested RT-PCR product, was sequenced. The sequence database search indicates that the sequence is homologous to the published sequence of Puumala viruses. The sequence similarities were ranged from 71% to 97%. The highest sequence similarity was 97% with Puumala virus Vranicam strain, and the lowest was 71% with Puumala virus K27 isolate. Puumala virus Vranicam strain was isolated from a bank vole (Clethrionomys glareolus) in Bosnia-Hercegovina. Puumala virus K27 was isolated from human in Russia. This analysis confirms that bats (Rhinolophus ferrum-equinum) in Korea are natural reservoir of Puumala virus.

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Effects of Guanidination with Trypsin, Lys-C, or Glu-C Digestion on Mass Spectrometric Signal Intensity and Protein Sequence Coverage

  • Han, Hye-Sun;Nho, Seon-Ho;Lee, Ae-Ra;Kim, Jeong-Kwon
    • Bulletin of the Korean Chemical Society
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    • v.31 no.6
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    • pp.1527-1534
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    • 2010
  • The conventional peptide modification process of guanidination, in which the amino groups of lysine residues are converted to guanidino groups using O-methylisourea to create more basic homoarginine residues, is often used to improve the signal intensity of lysine-containing peptides in matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). Here, we used three different protease enzymes (trypsin, Lys-C, and Glu-C) to evaluate the effects of guanidination on the MS signals of two enzymatically digested proteins. Horse heart myoglobin and bovine serum albumin were guanidinated either before or after digestion with trypsin, Lys-C, or Glu-C. The resulting peptides were subjected to MALDI-MS, and signal intensities and sequence coverage were systematically evaluated for each digest. Guanidination prior to Glu-C digestion improved sequence coverage for both proteins. For myoglobin, guanidination before enzymatic digestion with trypsin or Lys-C also enhanced sequence coverage, but guanidination after enzymatic digestion enhanced sequence coverage only with Lys-C. For albumin, guanidination either before or after Glu-C digestion increased sequence coverage, whereas pre- or post-digestion guanidination decreased sequence coverage with trypsin and Lys-C. The amino acid composition of a protein appears to be the major factor determining whether guanidination will enhance its MALDI-MS sequence coverage.

Inference and estimation using experience-based knowledge

  • Sakai, Y.
    • 제어로봇시스템학회:학술대회논문집
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    • 1992.10b
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    • pp.636-641
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    • 1992
  • In the human cognitive activity, experiencing plays a basic role. This is modeled by the idea of experience sequence here, which has been proposed by the author for the incorporation of the factor of experiencing in man-machine communication. Experience sequence is for modeling the human concept formation through experiencing. Knowledge manipulation requires concept understanding as its basis. An experience sequence deals with such a process of concept formation.

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The efficient implementation of the Hop Sequence in Bluetooth Baseband (블루투스 베이스밴드 홉 시퀀스의 효율적 구현 방안)

  • 김현미;김진숙;임재윤
    • Proceedings of the IEEK Conference
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    • 2002.06a
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    • pp.299-302
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    • 2002
  • This paper has been understanded the whole concept of hop sequence in bluetooth baseband and studied algorithm of its each type. It is implemented and verified both hardware and firmware of hop sequence according to Sepcification ver.1.1, and compared and examined two schemes with performance, size, cost, and time-to-market, etc. This paper is suggested that it is the efficient system division to design optimum system depending on developing environment.

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ITS2 DNA Sequence Analysis for Eight Species of Delphacid Planthoppers and a Loop-mediated Isothermal Amplification Method for the Brown Planthopper-specific Detection (멸구과 8종의 ITS2 DNA 염기서열 비교 분석과 고리매개등온증폭법(LAMP)을 이용한 벼멸구 특이 진단법)

  • Seo, Bo Yoon;Park, Chang Gyu;Koh, Young-Ho;Jung, Jin Kyo;Cho, Jumrae;Kang, Chanyeong
    • Korean journal of applied entomology
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    • v.56 no.4
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    • pp.377-385
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    • 2017
  • Estimates of evolutionary sequence divergence and inference of a phylogenetic tree for eight delphacid planthopper species were based on the full-length nucleotide sequence of the internal transcribed spacer 2 (ITS2) region. Size of the ITS2 DNA sequence varied from 550 bp in Sogatella furcifera to 699 bp in Nilaparvata muiri. Nucleotide sequence distance ($d{\pm}S.E.$) was lowest between N. muiri and N. bakeri ($0.001{\pm}0.001$), and highest between Ecdelphax cervina and Stenocranus matsumurai ($0.579{\pm}0.021$). Sequence distance between N. lugens and other planthoppers ranged from $0.056{\pm}0.008$ (N. muiri) to $0.548{\pm}0.021$ (S. matsumurai). In the neighbor-joining phylogenetic tree, all planthoppers were clustered separately into a species group, except N. muiri and N. bakeri. The ITS2 nucleotide sequence of N. lugens was used to design four loop-mediated isothermal amplification (LAMP) primer sets (BPH-38, BPH-38-1, BPH-207, and BPH-92) for N. lugens species-specific detection. After the LAMP reaction of three rice planthoppers, N. lugens, S. furcifera, and Laodelphax striatellus, with the four LAMP primer sets for 60 min at $65^{\circ}C$, LAMP products were observed in the genomic DNA of N. lugens only. In the BPH-92 LAMP primer set, the fluorescence relative to that of the negative control differed according to the amount of DNA (0.1 ng, 10 ng, and 100 ng) and incubation duration (20 min, 30 min, 40 min, and 60 min). At $65^{\circ}C$ incubation, the difference was clearly observed after 40 min with 10 ng and100 ng, but with a 60-min incubation period, the minimum DNA needed was 0.1 ng. However, there was little difference in fluorescence among all DNA amounts tested with 20 or 30 min incubations.

Molecular Authentication and Phylogenetic Relationship of Bupleurum Species by the rDNA-ITS Sequences (rDNA-ITS 염기서열 분석을 통한 시호 종 감별용 유전자 마커 개발 및 유연관계 분석)

  • Moon, Byeong-Cheol;Choo, Byeong-Kil;Ji, Yun-I;Yoon, Tae-Sook;Lee, A-Young;Cheon, Myeong-Sook;Kim, Bo-Bae;Kim, Ho-Kyoung
    • The Korea Journal of Herbology
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    • v.24 no.3
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    • pp.59-68
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    • 2009
  • Objectives : Bupleuri Radix (Siho) is prescribed as the root of different Bupleurum species on the pharmarcopoeia in Korea and China. Moreover, other species and varieties of the genus Bupleurum have been also distributed on the herbal market as Bupleuri Radix. However, due to the morphological similarity and frequent occurrence of intermediate forms, the correct identification of this radix is very difficult. To develop a reliable method for correct identification and improving the quality standards of official Bupleuri Radix, we analyzed sequences of the ribosomal RNA gene and internal transcribed spacer (rDNA-ITS) region. Methods : PCR amplification of rDNA-ITS region was performed using ITS1 and ITS4 primer from 6 Bupleurum species and 1 variety, B. falcatum L. (Siho), an improved breed of B. falcatum L. (Samdo-Siho), B. chinense DC. (Buk-Siho), B. scorzonerifolium Willd. (Nam-Siho), B. longiadiatum Turcz. (Gae-Siho), B. euphorbiodes Nakai (Deungdae-Siho) and B. latissimum Nakai (Seom-Siho), and nucleotide sequence was determined after sub-cloning into the pGEM-Teasy vector. Authentic marker nucleotides were estimated by the analysis of ClastalW using entire rDNA-ITS sequence of three samples per species. Results : In comparative analysis of the rDNA-ITS sequences, we found specific nucleotides to distinguish Korean (B. falcatum L. and its variety) and Chinese official species (B. chinense DC. and B. scorzonerifolium Willd.) from others at positions 411 and 447, and positions 89, 101, 415 and 599, respectively. Futhermore, we also found nucleotide indels (insertion and/or deletion) and substitutions to identify each of different Bupleurum species, 2 positions for B. falcatum L. and its variety, 6 positions for B. chinense DC., 49 positions for B. scorzonerifolium Willd., 8 positions for B. euphorbioides Nakai, 7 positions for B. longiradiatum Nakai and 9 positions for B. latissimum Nakai. These sequence differences at corresponding positions are avaliable nucleotide markers to determine the botanical origins of Bupleuri Radix. Moreover, we confirmed the phylogenetic relationship of B. latissimum Nakai, a Korean endemic speices, among Bupleurum species based on the rDNA-ITS sequence. Conclusions : These marker nucleotides would be useful to identify the official herbal medicines by the providing of definitive information that can identify each plant species and distinguish it from unauthentic adulterant Bupleurum species.