• Title/Summary/Keyword: Sequencing Strategy

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Monitoring of Bifenazate Resistant Two-spotted Spider Mite, Tetranychus urticae Using Molecular Detection Method (분자학적 진단방법을 이용한 bifenazate 저항성 점박이응애 모니터링)

  • Lee, Kyu-Ri;Shin, Yun-Ho;Cho, Sun-Ran;Koo, Hyun-Na;Choi, Jang-Jeon;Ahn, Ki-Su;Kim, Gil-Hah
    • The Korean Journal of Pesticide Science
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    • v.15 no.1
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    • pp.61-67
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    • 2011
  • In 2010, two-spotted spider mite, Tetranychus urticae was collected from the rose greenhouse and apple orchards in Cheongju (CJ), Chungju (CUJ)-1, CUJ-2, Kangjin (KJ), Yesan (YS), and Yeongju (YJ). Among them, KJ and YS strain showed high resistance to bifenazate of 964.5- and 1l30-fold, respectively. The other strains showed low resistance to bifenazate. By analyzing the mitochondrial cytochrome b (cytb) sequence, G126S point mutation was detected in KJ and YS strain. Thus, G126S point mutation in the mitochondrial cytb was available molecular detection marker for selection of bifenazate resistant T. urticae. Two molecular detection methods, quantitative sequencing (QS) and PCR amplification of specific alleles (PASA) were well detected specific G126S point mutation. Therefore, these methods can be used to monitor the resistance allele in field population of T. urticae and bifenazate resistance management strategy.

Current Status and Prospect of Wheat Functional Genomics using Next Generation Sequencing (차세대 염기서열분석을 통한 밀 기능유전체 연구의 현황과 전망)

  • Choi, Changhyun;Yoon, Young-Mi;Son, Jae-Han;Cho, Seong-Woo;Kang, Chon-Sik
    • Korean Journal of Breeding Science
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    • v.50 no.4
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    • pp.364-377
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    • 2018
  • Hexaploid wheat (common wheat/bread wheat) is one of the most important cereal crops in the world and a model for research of an allopolyploid plant with a large, highly repetitive genome. In the heritability of agronomic traits, variation in gene presence/absence plays an important role. However, there have been relatively few studies on the variation in gene presence/absence in crop species, including common wheat. Recently, a reference genome sequence of common wheat has been fully annotated and published. In addition, advanced next-generation sequencing (NGS) technology provides high quality genome sequences with continually decreasing NGS prices, thereby dawning full-scale wheat functional genomic studies in other crops as well as common wheat, in spite of their large and complex genomes. In this review, we provide information about the available tools and methodologies for wheat functional genomics research supported by NGS technology. The use of the NGS and functional genomics technology is expected to be a powerful strategy to select elite lines for a number of germplasms.

A Study on Environmental Design Method based on Open Narrative Structure - A Case of Designing of Arirang Culture Park - (열린 내러티브 구조를 이용한 환경설계 방법 연구 - 용산 아리랑 문화공원을 설계사례로 -)

  • 이상경;조경진
    • Journal of the Korean Institute of Landscape Architecture
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    • v.31 no.2
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    • pp.12-27
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    • 2003
  • The purpose of this study is to look for an environmental design method based on open narrative structure, and to promote various experiences and interpretations of space through user's engagements. That is to say, the designer does not lead specific events through separate Boning but using the continuous open composition users are provided with margins for their imaginations. Spatial formulation through open narrative structure gives us various thoughts and it plays an important role in making a sequential space. Like an abstract painting, it is a complex story making or arranging a montage of images containing stories that elicit the reader's engagement through diverse interpretations. Like this, open composition exists in an ambiguous state and it is possible to interpret unfinished‘evolving work’within it. Utilizing open narrative structure, this study attempts to apply the idea of sequencing and open composition in the case of designing Arirang Culture Park. Open composition should induce various engagements by users and could be a medium which organically connects nature, culture and people. The spatial strategies of‘ambiguity’ and‘transparency’are like a bundle of complex and heterogeneous factors. Finally, the study focuses on the ‘integration’of the main ideas that compose multilayered space. ‘Voidness’and‘thickening’are also used for spatial strategies in open narrative structure. As alternative plans for undecided programs of the space, the voidness can be a strategic design program with flexibility about changes of futures. Also, thickening can be a strategic design program for functional reinforcement of the space, for the dramatic effects and for the generation of incidental events. Although both voidness and thickening seem paradoxical, we can see they are similar in the way that both focus on various spatial uses and by how they do not function as one-to-one correspondence, but as multiple correspondences. Therefore, open narrative structure is possible to apply in designing space and it can be an alternative design strategy for inducing multiple interpretations of space.

Effects of Nitrogen and Phosphorus Starvation on Growth and Fatty Acid Production in Newly Isolated Two Freshwater Green Microalgae from Nakdonggang River (낙동강 수계에서 분리한 녹조류 2종의 질소와 인의 결핍에 따른 생장 및 지방산 변화 연구)

  • Yim, Kyung June;Park, Hanwool;Lee, Chang Soo;Jo, Bok Yeon;Nam, Seung Won;Lee, Choul-Gyun;Kim, Z-Hun
    • Journal of Marine Bioscience and Biotechnology
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    • v.11 no.2
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    • pp.81-88
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    • 2019
  • In this study, effects of nitrogen (N) and phosphorus (P) starvation on the cell growth and fatty acid (FA) production of newly isolated freshwater microalgae were investigated. The microalgae were identified as Chlorella sp. and Parachlorella sp. through 18S rRNA sequencing. Optimal culture temperature and light intensity were investigated using a high-throughput photobioreator, and the result was validated in 0.5 L bubble column photobioreactors using BG-11 without NaNO3 and/or K2HPO4. Under nutrient starvation conditions, total FA contents of the microalgae were significantly changed rather than FA composition. Starvation of both N and P was most effective for increasing FA contents in Parachlorella sp (24.4±0.1%) whereas highest FA contents (42.6±1.8%) was achieved when only P was starved in Chlorella sp. among tested conditions. These results suggest an effective strategy for increasing FA production from microalgae using appropriate nutrient starvation.

Current status and prospects of genomics and bioinformatics in grapes (포도 유전체 연구현황 및 전망)

  • Hur, Youn Young;Jung, Sung Min;Yun, Hae Keun
    • Journal of Plant Biotechnology
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    • v.42 no.4
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    • pp.298-311
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    • 2015
  • Grape is one of the important fruit crops around the world, and exposed to disease and pests, and internal or environmental stresses in the vineyards. Breeding and cultivation of new varieties of high quality-grapes resistant to diseases and pests and tolerant to stresses are the most important steps in the grape production. However, conventional breeding has laborious and time-consuming procedures in maintaining and selecting seedlings in the fields. Development of molecular breeding technology through understanding of molecular mechanism of useful traits can be used as an alternative strategy to improve the efficiency of grape breeding program by cross hybridization in grape development programs. The completion of the grape genome sequencing project provided the way to discover the novel genes and to analyze their functions. Comparative genomics, transcriptomic analysis, and the genome-wide identification and analysis of useful genes as well as development of molecular marker for valuable traits could provide novel insights into fruit quality and the responses to diseases and stresses, and can be used as important information in molecular breeding programs for grape development.

Scolopendrasin I: a novel antimicrobial peptide isolated from the centipede Scolopendra subspinipes mutilans

  • Lee, Joon Ha;Kim, In-Woo;Kim, Mi-Ae;Yun, Eun-Young;Nam, Sung-Hee;Ahn, Mi-Young;Lee, Young Bo;Hwang, Jae Sam
    • International Journal of Industrial Entomology and Biomaterials
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    • v.31 no.1
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    • pp.14-19
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    • 2015
  • In a previous report, we identified several candidate antimicrobial peptides through de novo RNA sequencing of the centipede Scolopendra subspinipes mutilans. Here, we identify and characterize one of these peptides, Scolopendrasin I. We identified the centipede antimicrobial peptide Cecropin from the centipede transcriptome using an SVM algorithm, and subsequently analyzed the amino acid sequence for predicted secondary structure using a GOR algorithm. We identified an alpha helical region of Cecropin and named it Scolopendrasin I. We then assessed antimicrobial and hemolytic activity of Scolopendrasin I. Scolopendrasin I showed antimicrobial activity against various microbes, including antibiotic-resistant Gram-negative bacteria, in a radial diffusion assay. Scolopendrasin I had potent antibacterial activity against acne-associated microbes in a colony count assay and showed no hemolytic activity in a hemolysis assay. In addition, we confirmed that Scolopendrasin I bound to the surface of bacteria via a specific interaction with lipoteichoic acid and lipopolysaccharide, two components of bacterial cell membranes. In conclusion, the results presented here provide evidence that this is an efficient strategy for antimicrobial peptide candidate identification and that Scolopendrasin I has potential for successful antibiotic development.

Novel Preparation and Characterization of the α4-loop-α5 Membrane-perturbing Peptide from the Bacillus thuringiensis Cry4Ba δ-endotoxin

  • Leetachewa, Somphob;Katzenmeier, Gerd;Angsuthanasombat, Chanan
    • BMB Reports
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    • v.39 no.3
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    • pp.270-277
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    • 2006
  • Helices 4 and 5 of the Bacillus thuringiensis Cry4Ba $\delta$-endotoxin have been shown to be important determinants for mosquito-larvicidal activity, likely being involved in membrane-pore formation. In this study, the Cry4Ba mutant protein containing an additional engineered tryptic cleavage site was used to produce the $\alpha4$-$\alpha5$ hairpin peptide by an efficient alternative strategy. Upon solubilization of toxin inclusions expressed in Escherichia coli and subsequent digestion with trypsin, the 130-kDa mutant protoxin was processed to protease-resistant fragments of ca. 47, 10 and 7 kDa. The 7-kDa fragment was identified as the $\alpha4$-loop-$\alpha5$ hairpin via N-terminal sequencing and mass spectrometry, and was successfully purified by size-exclusion FPLC and reversed-phase HPLC. Using circular dichroism spectroscopy, the 7-kDa peptide was found to exist predominantly as an $\alpha$-helical structure. Membrane perturbation studies by using fluorimetric calcein-release assays revealed that the 7-kDa helical hairpin is highly active against unilamellar liposomes compared with the 65-kDa activated full-length toxin. These results directly support the role of the $\alpha4$-loop-$\alpha5$ hairpin in membrane perturbation and pore formation of the full-length Cry4Ba toxin.

Immunogenomics approaches to study host innate immunity against intestinal parasites

  • Lillehoj, Hyun S.
    • Proceedings of the Korea Society of Poultry Science Conference
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    • 2006.11a
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    • pp.7-16
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    • 2006
  • Poultry products including meat and eggs constitute a major protein source in the American diet and disease - causing pathogens represent major challenges to the poultry industry. More than 95 % of pathogens enter the host through the mucosal surfaces of the respiratory, digestive and reproductive tracts and over the past few decades, the two main mechanisms used to control diseases have been the use of vaccines and antibiotics. However, in the poultry industry, there are mounting concerns over the ability of current vaccines to adequately protect against emerging hyper - virulent strains of pathogens and a lack of suitable, cost effective adjuvants. Thorough investigation of the immunogenetic responses involved in host-pathogen interactions will lead to the development of new and effective strategies for improving poultry health, food safety and the economic viability of the US poultry industry. In this paper, I describe the development of immunogenomic and proteomic tools to fundamentally determine and characterize the immunological mechanisms of the avian host to economically significant mucosal pathogens such as Eimeria. Recent completion of poultry genome sequencing and the development of several tissue-specific cDNA libraries in chickens are facilitating the rapid application of functional immunogenomics in the poultry disease research. Furthermore, research involving functional genomics, immunology and bioinformatics is providing novel insights into the processes of disease and immunity to microbial pathogens at mucosal surfaces. In this presentation, a new strategy of global gene expression using avian macrophage (AMM) to characterize the multiple pathways related to the variable immune responses of the host to Eimeria is described. This functional immunogenomics approach will increase current understanding of how mucosal immunity to infectious agents operates, and how it may be enhanced to enable the rational development of new and effective strategies against coccidiosis and other mucosal pathogens.

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Development and Effect of Learning Materials of Earth Science Using Simplifying Condition Method (단순화 조건법을 이용한 지질 연대 분야의 학습 자료 개발과 그 효과)

  • Kim, Jong-Hee;Jeong, Hui-Gyeong;Kim, Sang-Dal
    • Journal of the Korean earth science society
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    • v.24 no.6
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    • pp.495-507
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    • 2003
  • The purpose of this study was three-folded to suggest the Simplifying Conditions Method (SCM) as a means of task analysis and sequencing of instructional content, to develop teaching-learning materials by analyzing part of the geological time scale of the earth science and finally to analyze the effectiveness of this method. SCM began by simplifying a complex task into the basic components by eliminating various complexities, which produced a simple representative of the entire task. The next step was to relax conditions on the basic version one by one, thereby gradually introducing progressively more complex tasks to the students. This sequential strategy enabled students to understand the task holistically and to acquire authentic skills from very onset of the course. Moreover, Early mastery of skills enhances the effectiveness and efficiency of instruction. The result of this study revealed that instruction through SCM was more effective in developing students' self-directed learning characteristics and academic achievement than instruction through sequential task analysis methodology.

A New ColE1-like Plasmid Group Revealed by Comparative Analysis of the Replication Proficient Fragments of Vibrionaceae Plasmids

  • Pan, Li;Leung, P.C.;Gu, Ji-Dong
    • Journal of Microbiology and Biotechnology
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    • v.20 no.8
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    • pp.1163-1178
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    • 2010
  • Plasmids play important roles in horizontal gene transfer among Vibrionaceae, but surprisingly little is known about their replication and incompatibility systems. In this study, we successfully developed a bioinformatics-assisted strategy of experimental identification of seven Vibrio plasmid replicons. Comparative sequences analysis of the seven Vibrio plasmid replicons obtained in this study together with eight published Vibrionaceae plasmid sequences revealed replication-participating elements involved in the ColE1 mode of replication initiation and regulation. Like plasmid ColE1, these Vibrionaceae plasmids encode two RNA species (the primer RNA and the antisense RNA) for replication initiation and regulation, and as a result, the 15 Vibrionaceae plasmids were designated as ColE1-like Vibrionaceae (CLV) plasmids. Two subgroups were obtained for the 15 CLV plasmids, based on comparison of replicon organization and phylogenetic analysis of replication regions. Coexistence of CLV plasmids were demonstrated by direct sequencing analysis and Southern hybridization, strongly suggesting that the incompatibility of CLV plasmids is determined mainly by the RNA I species like the ColE1-like plasmids. Sequences resembling the conserved Xer recombination sites were also identified on the CLV plasmids, indicating that the CLV plasmids probably use the host site-specific recombination system for multimer resolution like that used by ColE1-like plasmids. All the results indicated that the 15 plasmids form a new ColE1-like group, providing a basis for the rapid characterization and classification of Vibrionaceae plasmids.