• Title/Summary/Keyword: Sequence Analyses

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Improved Methodology for Identification of Cryptomonads: Combining Light Microscopy and PCR Amplification

  • Xia, Shuang;Cheng, Yingyin;Zhu, Huan;Liu, Guoxiang;Hu, Zhengyu
    • Journal of Microbiology and Biotechnology
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    • v.23 no.3
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    • pp.289-296
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    • 2013
  • Cryptomonads are unicellular, biflagellate algae. Generally, cryptomonad cells cannot be preserved well because of their fragile nature, and an improved methodology should be developed to identify cryptomonads from natural habitats. In this study, we tried using several cytological fixatives, including glutaraldehyde, formaldehyde, and their combinations to preserve field samples collected from various waters, and the currently used fixative, Lugol's solution was tested for comparison. Results showed that among the fixatives tested, glutaraldehyde preserved the samples best, and the optimal concentration of glutaraldehyde was 2%. The cell morphology was well preserved by glutaraldehyde. Cells kept their original color, volume, and shape, and important taxonomic features such as furrow/gullet complex, ejectosomes, as well as flagella could be observed clearly, whereas these organelles frequently disappeared in Lugol's solution preserved samples. The osmotic adjustments and buffers tested could not preserve cell density significantly higher. Statistical calculation showed the cell density in the samples preserved by 2% glutaraldehyde remained stable after 43 days of the fixation procedure. In addition, DNA was extracted from glutaraldehyde preserved samples by grinding with liquid nitrogen and the 18S rDNA sequence was amplified by PCR. The sequence was virtually identical to the reference sequence, and phylogenetic analyses showed very close relationship between it and sequences from the same organism. To sum up, the present study demonstrated that 2% unbuffered glutaraldehyde, without osmotic adjustments, can preserve cryptomonads cells for identification, in terms of both light microscopy and phylogenetic analyses based on DNA sequences.

First Report of Cucumber mosaic virus Infecting Pinewood Coneflower (Rudbeckia bicolor) in Korea

  • Kim, Mi-Kyeong;Kwak, Hae-Ryun;Ko, Sug-Ju;Lee, Su-Heon;Kim, Jeong-Soo;Kim, Kook-Hyung;Cha, Byeong-Jin;Choi, Hong-Soo
    • The Plant Pathology Journal
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    • v.26 no.1
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    • pp.93-98
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    • 2010
  • A virus isolate causing symptoms of yellow mosaic, fern leaves, malformation and plant necrosis on Rudbeckia bicolor was prevalent around Pyeongchang area in Korea. The causal virus was identified as Cucumber mosaic virus (CMV) using characteristics from biological, serological and molecular analyses and named as CMV-Rb. CMV-Rb caused mosaic on Nicotiana benthamiana, N. tabacum, Capsicum annuum, and Lycopersicon esculentum. However, typical local lesions did not develop on inoculated Pisum sativum, Cucurbita moschata, Datura stramonium and Tetragonia expansa plants. Full-length genome sequences of CMV-Rb RNAs 1, 2 and 3 were obtained using 12 primer pairs by RT-PCR analysis. The genome of CMV-Rb RNA segments 1, 2, and 3 consists of 3363nt, 3049nt, and 2214nt in length, respectively. In order to ascertain their taxonomic identity, nucleotide and the deduced amino acid sequence analyses RNAs 1, 2 and 3 of CMV-Rb isolates were conducted with previously reported sequences of CMV strains and/or isolates. CMV-Rb RNAs showed about 90 to 99% sequence identity to those of subgroup I strains suggesting that CMV-Rb is more closely related to CMV isolates belong to subgroup I. To our knowledge, this is the first report of CMV on Rudbeckia bicolor in Korea.

Genetic Diversity Among Pseudomonas syringae pv. morsprunorum Isolates from Prunus mume in Korea and Japan by Comparative Sequence Analysis of 16S rRNA Gene

  • Lee, Young-Sun;Koh, Hyun-Seok;Sohn, San-Ho;Koh, Young-Jin;Jung, Jae-Sung
    • The Plant Pathology Journal
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    • v.28 no.3
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    • pp.295-298
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    • 2012
  • Genetic diversity among Pseudomonas syringae pv. morsprunorum isolates from Prunus mume in Korea and Japan was investigated by comparative sequence analysis of the 16S rRNA gene. The strains included 24 field isolates recovered from P. mume in Korea along with seven Japanese strains. Two strains isolated from P. salicina in Japan, one strain from P. avium in the United Kingdom, and the pathotype strain were also used for comparison with their 16S rRNA gene sequences. Nearly complete 16S rRNA gene sequences were sequenced in all 35 strains, and three sequence types, designated types I, II and III, were identified. Eleven strains consisting of five Korean isolates, five Japanese strains, and one strain from the United Kingdom belonged to type I, whereas the pathotype strain and another 19 Korean isolates belonged to type III. Another four Japanese strains belonged to type II. Type I showed 98.9% sequence homology with type III. Type I and II had only two heterogeneous bases. The 16S rRNA sequence types were correlated with the races of P. syringae pv. morsprunorum. Type I and II strains belonged to race 1, whereas type III isolates were included in race 2. Sequence analyses of the 16S rRNA gene from P. syringae pv. morsprunorum were useful in identifying the races and can further be used for epidemiological surveillance of this pathogen.

Complete Genome Sequence Analysis of Two Divergent Groups of Sweet potato chlorotic fleck virus Isolates Collected from Korea

  • Kwak, Hae-Ryun;Kim, Jaedeok;Kim, Mikyeong;Seo, Jang-Kyun;Kim, Jeong-Soo;Choi, Hong-Soo
    • The Plant Pathology Journal
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    • v.34 no.5
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    • pp.451-457
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    • 2018
  • The Sweet potato chlorotic fleck virus (SPCFV), of the genus Carlavirus (family Betaflexiviridae), was first detected as one of several viruses infecting sweet potatoes (Ipomea batatas L.) in Korea. Out of 154 sweet potato samples collected in 2012 that were showing virus-like symptoms, 47 (31%) were infected with SPCFV, along with other viruses. The complete genome sequences of four SPCFV isolates were determined and analyzed using previously reported genome sequences. The complete genomes were found to contain 9,104-9,108 nucleotides, excluding the poly-A tail, containing six putative open reading frames (ORFs). Further, the SPCFV Korean isolates were divided into two groups (Group I and Group II) by phylogenetic analysis based on the complete nucleotide sequences; Group I and Group II had low nucleotide sequence identities of about 73%. For the first time, we determined the complete genome sequence for the Group II SPCFV isolates. The amino acid sequence identity in coat proteins (CP) between the two groups was over 90%, whereas the amino acid sequence identity in other proteins was less than 80%. In addition, SPCFV Korean isolates had a low amino acid sequence identity (61% CPs and 47% in the nucleotide-binding protein [NaBp] region) to that of Melon yellowing-associated virus (MYaV), a typical Carlavirus.

Taxonomic revision of the genus Herposiphonia (Rhodomelaceae, Rhodophyta) from Korea, with the description of three new species

  • Koh, Young Ho;Kim, Myung Sook
    • ALGAE
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    • v.33 no.1
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    • pp.69-84
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    • 2018
  • We examined the species diversity of Herposiphonia on Korean coasts, based on a combination of morphology and molecular analyses of the mitochondrial COI-5P DNA barcode marker and plastid rbcL gene. We report the presence of eight species including three novel species: H. donghaensis sp. nov., H. jejuinsula sp. nov., H. sparsa sp. nov., H. caespitosa, H. fissidentoides, H. insidiosa, H. parca, and H. subdisticha. Specimens were separated into eight clades in both the COI-5P and rbcL gene analyses, with 1.3-19.6 and 6.6-15% interspecific sequence divergence, respectively. These eight species are also distinguishable by several morphological characteristics such as: branching pattern (d/i pattern in H. donghaensis sp. nov. and H. sparsa sp. nov.; d/d/d/i pattern in others), shape of determinate branch (ligulate in H. fissidentoides; terete in others), number of vegetative trichoblasts (1-2 in H. insidiosa and H. sparsa sp. nov.; 3-4 in H. caespitosa; absent in others), and number of segments and pericentral cells in determinate branches. About three novel species revealed by our analyses, H. donghaensis sp. nov. is newly discovered, and H. jejuinsula sp. nov. and H. sparsa sp. nov. were previously reported in Korea as H. nuda and H. secunda, respectively. Our results show that DNA barcoding and rbcL analyses are useful for delimiting species boundaries and discovering cryptic species diversity in the genus Herposiphonia.

The Cell Resequencing Buffer for the Cell Sequence Integrity Guarantee for the Cyclic Banyan Network (사이클릭 벤얀 망의 셀 순서 무결성 보장을 위한 셀 재배열 버퍼)

  • 박재현
    • Journal of the Institute of Electronics Engineers of Korea TC
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    • v.41 no.9
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    • pp.73-80
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    • 2004
  • In this paper, we present the cell resequencing buffer to solve the cell sequence integrity problem of the Cyclic banyan network that is a high-performance fault-tolerant cell switch. By offering multiple paths between input ports and output ports, using the deflection self-routing, the Cyclic banyan switch offer high reliability, and it also solves congestion problem for the internal links of the switch. By the way, these multiple paths can be different lengths for each other. Therefore, the cells departing from an identical source port and arriving at an identical destination port can reach to the output port as the order that is different from the order arriving at input port. The proposed cell resequencing buffer is a hardware sliding window mechanism. to solve such cell sequence integrity problem. To calculate the size of sliding window that cause the prime cost of the presented device, we analyzed the distribution of the cell delay through the simulation analyses under traffic load that have a nonuniform address distribution that express tile Property of traffic of the Internet. Through these analyses, we found out that we can make a cell resequencing buffer by which the cell sequence integrity is to be secured, by using a, few of ordinary memory and control logic. The cell resequencing buffer presented in this paper can be used for other multiple paths switching networks.

Seismic Stratigraphy and Sedimentary Environment of the Dukjuk-Do Sand Ridge in Western Gyeonggi Bay, Korea (경기만 서부 덕적도 사퇴의 탄성파층서 및 퇴적환경 연구)

  • Lee, Yoon-Oh;Choi, Sang-Il;Jeong, Gyo-Cheol
    • The Journal of Engineering Geology
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    • v.24 no.1
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    • pp.9-21
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    • 2014
  • We examined high-resolution seismic data, side scan sonar data, surface sediments, and vibrocore samples from a sand ridge off the western part of Dukjuk-Do in Gyeonggi Bay, with the aim of interpretation of seismic stratigraphy and sedimentary environment. Based on the seismic data, the deposited sands are divided into three sedimentary units. 14C age data indicate that the top sequence (sequence I) formed at 5000-6000 yr BP, when a transgression resulted in strong shifting tides. Analyses of the vibrocore samples indicate that sequence II is a paleo-mudflat layer of intertidal sediments dominated by mud. Sequence III consists of terrestrial sediments that are presumed to have been deposited at the end of the Pleistocene, unconformably overlying the acoustic bedrock and Mesozoic granite. The side scan sonar data indicate that sand waves were formed on the seabed on top of the sand ridge. Generally, this is the direction of $N20^{\circ}E$, which coincides with the direction of tidal flow. Sand ripples occur away from the top of the sand ridge and are distributed homogeneously across a sandy slope. Vibrocore analyses indicate that the surface sediments and core sediments (samples VC-1, -2, and -3) are homogeneous, without any internal structures, and are characterized by a mixture of medium and fine sand (1-$2{\phi}$), respectively.

Flutter stability of a long-span suspension bridge during erection

  • Han, Yan;Liu, Shuqian;Cai, C.S.;Li, Chunguang
    • Wind and Structures
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    • v.21 no.1
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    • pp.41-61
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    • 2015
  • The flutter stability of long-span suspension bridges during erection can be more problematic and more susceptible to be influenced by many factors than in the final state. As described in this paper, numerical flutter stability analyses were performed for the construction process of Zhongdu Bridge over Yangtze River using the commercial FE package ANSYS. The effect of the initial wind attack angle, the sequence of deck erection, the stiffness reduction of stiffening girders, the structural damping, and the cross cables are discussed in detail. It was found that the non-symmetrical sequence of deck erection was confirmed to be aerodynamically favourable for the deck erection of long-span suspension bridges and the best erection sequence should be investigated in the design phase. While the initial wind attack angle of $-3^{\circ}$ is advantageous for the aerodynamic stability, $+3^{\circ}$ is disadvantageous compared with the initial wind attack angle of $0^{\circ}$ during the deck erection. The stiffness reduction of the stiffening girders has a slight effect on the flutter wind speed of the suspension bridge during erection, but structural damping has a great impact on it, especially for the early erection stages.

Nucleotide Sequence of Rat Transketolase and Liver-Specific Pretranslational Activation During Postnatal Development

  • Kim, Sung-Min F.;Kim, Byung-Moon;Jeng, Jingjau;Soh, Yun-Jo;Bak, Choong-Il;Huh, Jae-Wook;Song, Byoung-J.
    • BMB Reports
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    • v.29 no.2
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    • pp.146-150
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    • 1996
  • A 2.1 kb cDNA clone for rat transketolase was isolated from rat liver ${\lambda}gt11$ cDNA library and its sequence was determined. The predicted rat transketolase (655 amino acids with $M_r$ 71,186) is highly similar (92%) to that of the human enzyme except that it contains an extra 32 amino acids at its N-terminus. Although it is less similar (<27%) to transketolases from non-mammalian species, the functional motifs such as the catalytic sites and thiamine binding domain are well conserved in the rat enzyme. Southern blot analysis of genomic DNA verified that transketolase appears to be derived from a single gene. Immunoblot and Northern blot analyses suggested that hepatic transketolase was activated pretranslationally by a 2.1-fold while little change was observed in brain enzyme, indicating a tissue-specific pretranslational activation during postnatal development.

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Taxonomic Position of Korean Isolates of Rhizoctonia solani Based on RAPD and ITS Sequencing of Ribosomal DNA

  • Jeon, Young-Ah;Kim, Wan-Gyu;Kim, Dae-Ho;Kwon, Soon-Wo;Hong, Seung-Beom
    • The Plant Pathology Journal
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    • v.26 no.1
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    • pp.83-89
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    • 2010
  • Taxonomic position of 46 Korean isolates of Rhizoctonia solani which were classified into nine intraspecific groups by anastomosis and cultural characteristics was analyzed by randomly amplified polymorphic DNA (RAPD) and sequence analyses of the internal transcribed spacer (ITS) regions of ribosomal DNA. All the isolates within each group showed highly similar band patterns in RAPD. The ITS regions of the isolates within the same groups showed a high level of sequence similarity above 96.0% whereas similarities among different groups were below 94.4%. When compared with several reference strains of R. solani from foreign countries, all the Korean isolates were clustered with the foreign isolates belonging to the same groups in the phylogenetic tree. All six Korean strains of AG-4 were identified as HG-1 out of 3 subgroup of AG-4. We discussed taxonomic position of Korean isolates of R. solani and showed that sequence analysis with ITS regions could be a rapid and useful method for identification of intraspecific group of R. solani.